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Sample records for human excreta urine

  1. Use of Human Excreta as Manure in Rural China

    Institute of Scientific and Technical Information of China (English)

    LIU Ying; HUANG Ji-kun; Precious Zikhali

    2014-01-01

    Empirical research has shown that the use of manure signiifcantly improves crop yield, soil fertilityand water and moisture conservation. Despite these documented benefits, however, there is a concern on the downward trend of manure use in agriculture in China. This paper examines factors contributing to this downward trend, with a particular focus on human excreta used in agriculture. Empirical analysis based on data from stratiifed random sampling of rural households in ifve provinces of China shows that about 85% of human excreta was still used as manure in agriculture in 2007 which was less than a decade ago when nearly all human excreta was used as manure. Econometric results suggest that income growth, rising population density and improvement in rural transportation signiifcantly contribute to declining use of human excreta as manure in agriculture. These results imply that the current downward trend will continue given China’s rising economic growth, urbanization and rural infrastructural improvement.

  2. Composition of human excreta - a case study from Souther Thailand

    DEFF Research Database (Denmark)

    Schouw, Nanette L..; Danteravanich, S.; Mosbæk, Hans

    2002-01-01

    . In the present study the composition of human. excreta has been studied in three case study areas in Southern Thailand: Kuan Lang, Phattalung and Prik. The inhabitants of the three areas represent people of Southern Thailand by age, sex, occupation, religion and type of residence. Human excreta was collected....... Furthermore, there was no significant influence of age, sex, occupation or religion on the chemical composition. The only significant variation was that the older people excreted larger amounts of total wet matter than the younger, which could be due to a higher water intake, in order to reduce the risk...

  3. Environmental impact of recycling nutrients in human excreta to agriculture compared with enhanced wastewater treatment

    Energy Technology Data Exchange (ETDEWEB)

    Spångberg, J. [Swedish University of Agricultural Sciences, Department of Energy and Technology, Box 7032, 750 07 Uppsala (Sweden); Tidåker, P. [Swedish Institute of Agricultural and Environmental Engineering, P.O. Box 7033, 750 07 Uppsala (Sweden); Jönsson, H., E-mail: hakan.jonsson@slu.se [Swedish University of Agricultural Sciences, Department of Energy and Technology, Box 7032, 750 07 Uppsala (Sweden)

    2014-09-15

    Human excreta are potential sources of plant nutrients, but are today usually considered a waste to be disposed of. The requirements on wastewater treatment plants (WWTPs) to remove nitrogen and phosphorus are increasing and to meet these requirements, more energy and chemicals are needed by WWTPs. Separating the nutrient-rich wastewater fractions at source and recycling them to agriculture as fertiliser is an alternative to removing them at the WWTP. This study used life cycle assessment methodology to compare the environmental impact of different scenarios for recycling the nutrients in the human excreta as fertiliser to arable land or removing them in an advanced WWTP. Three scenarios were assessed. In blackwater scenario, blackwater was source-separated and used as fertiliser. In urine scenario, the urine fraction was source-separated and used as fertiliser and the faecal water treated in an advanced WWTP. In NP scenario, chemical fertiliser was used as fertiliser and the toilet water treated in an advanced WWTP. The emissions from the WWTP were the same for all scenarios. This was fulfilled by the enhanced reduction in the WWTP fully removing the nutrients from the excreta that were not source-separated in the NP and urine scenarios. Recycling source-separated wastewater fractions as fertilisers in agriculture proved efficient for conserving energy and decreasing global warming potential (GWP). However, the blackwater and urine scenarios had a higher impact on potential eutrophication and potential acidification than the WWTP-chemical fertiliser scenario, due to large impacts by the ammonia emitted from storage and after spreading of the fertilisers. The cadmium input to the arable soil was very small with urine fertiliser. Source separation and recycling of excreta fractions as fertiliser thus has potential for saving energy and decreasing GWP emissions associated with wastewater management. However, for improved sustainability, the emissions from storage and

  4. Community perceptions of human excreta as fertilizer in peri-urban agriculture in Ghana.

    Science.gov (United States)

    Mariwah, Simon; Drangert, Jan-Olof

    2011-08-01

    Although human excreta contain the necessary nutrients for plant growth, local authorities in Ghana spend huge sums of money to dispose them as waste. Reusing excreta for agricultural purposes saves expenditure for chemical fertilizers, improves soil fertility, reduces poverty and ensures food security. People's attitudes and perceptions about excreta vary between cultures and even within specific cultures. This study aimed to explore attitudes and perceptions among a peri-urban agricultural community towards sanitized human excreta and its use. The study adopted an exploratory design and collected data from 154 randomly selected households using questionnaires and focus group discussions. It was found that there is a general negative attitude to fresh excreta and the handling of it. However, the residents accept that excreta can be used as fertilizer, but they are not willing to use it on their own crops or consume crops fertilized with excreta. The study recommends open discussions in the community for a successful implementation of ecological sanitation.

  5. Hygiene versus fertiliser the use of human excreta in agriculture - a Vietnamese example

    DEFF Research Database (Denmark)

    Jensen, Peter Kjær Mackie; Phuc, Pham Duc; Knudsen, Line Gram;

    2008-01-01

    The use of human excreta as fertiliser in agriculture is a common practice in parts of South East Asia benefiting production but at the same time a risk factor for increased helminth infections. This paper describes the hygienic handling of human excreta for use in agriculture in Central Vietnam...

  6. The human urine metabolome.

    Directory of Open Access Journals (Sweden)

    Souhaila Bouatra

    Full Text Available Urine has long been a "favored" biofluid among metabolomics researchers. It is sterile, easy-to-obtain in large volumes, largely free from interfering proteins or lipids and chemically complex. However, this chemical complexity has also made urine a particularly difficult substrate to fully understand. As a biological waste material, urine typically contains metabolic breakdown products from a wide range of foods, drinks, drugs, environmental contaminants, endogenous waste metabolites and bacterial by-products. Many of these compounds are poorly characterized and poorly understood. In an effort to improve our understanding of this biofluid we have undertaken a comprehensive, quantitative, metabolome-wide characterization of human urine. This involved both computer-aided literature mining and comprehensive, quantitative experimental assessment/validation. The experimental portion employed NMR spectroscopy, gas chromatography mass spectrometry (GC-MS, direct flow injection mass spectrometry (DFI/LC-MS/MS, inductively coupled plasma mass spectrometry (ICP-MS and high performance liquid chromatography (HPLC experiments performed on multiple human urine samples. This multi-platform metabolomic analysis allowed us to identify 445 and quantify 378 unique urine metabolites or metabolite species. The different analytical platforms were able to identify (quantify a total of: 209 (209 by NMR, 179 (85 by GC-MS, 127 (127 by DFI/LC-MS/MS, 40 (40 by ICP-MS and 10 (10 by HPLC. Our use of multiple metabolomics platforms and technologies allowed us to identify several previously unknown urine metabolites and to substantially enhance the level of metabolome coverage. It also allowed us to critically assess the relative strengths and weaknesses of different platforms or technologies. The literature review led to the identification and annotation of another 2206 urinary compounds and was used to help guide the subsequent experimental studies. An online database

  7. The fear of awful smell: risk perceptions among farmers in Vietnam using wastewater and human excreta in agriculture

    DEFF Research Database (Denmark)

    Knudsen, Line G; Phuc, Pham D; Hiep, Nguyen T

    2008-01-01

    Vietnamese farmers' health-risk awareness, knowledge, and practices related to their use of wastewater and human excreta was investigated in an anthropological study by a multidisciplinary team in peri-urban Hanoi and Nghe An Province. Farmers identified health risks associated with their use of ...

  8. Fertilizer and sanitary quality of digestate biofertilizer from the co-digestion of food waste and human excreta.

    Science.gov (United States)

    Owamah, H I; Dahunsi, S O; Oranusi, U S; Alfa, M I

    2014-04-01

    This research was aimed at assessing the fertilizer quality and public health implications of using digestate biofertilizer from the anaerobic digestion of food wastes and human excreta. Twelve (12) kg of food wastes and 3kg of human excreta were mixed with water in a 1:1 w/v to make 30-l slurry that was fed into the anaerobic digester to ferment for 60days at mesophilic temperature (22-31°C). Though BOD, COD, organic carbon and ash content in the feedstock were reduced after anaerobic digestion by 50.0%, 10.6%, 74.3% and 1.5% respectively, nitrogen, pH and total solids however increased by 12.1%, 42.5% and 12.4% respectively. The C/N ratios of the feedstock and compost are 135:1 and 15.8:1. The residual total coliforms of 2.10×10(8)CFU/100ml in the digestate was above tolerable limits for direct application on farmlands. Microbial analysis of the digestate biofertilizer revealed the presence of Pseudomonas, Klebsiella, Clostridium, Bacillus, Bacteroides, Penicillum, Salmollena, and Aspergillus. Klebsiella, Bacillus, Pseudomonas, Penicillum and Aspergillus can boost the efficiency of the biofertilizer through nitrogen fixation and nutrient solubility in soils but Klebsiella again and Salmollena are potential health risks to end users. Further treatment of the digestate for more efficient destruction of pathogens is advised. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Challenges of adoption of urine-diversion dry toilets technology as ...

    African Journals Online (AJOL)

    user

    Key words: Ecological sanitation, urine-diversion dry toilets, adoption, community participation, Tanzania. ... than attempting to control it, and uses the safe products of sanitized human excreta ...... Ecological Sanitation; Department for Natural.

  10. Bisphenol A levels in human urine.

    OpenAIRE

    Matsumoto, Akiko; Kunugita, Naoki; Kitagawa, Kyoko; Isse, Toyohi; Oyama, Tsunehiro; Foureman, Gary L; Morita, Masatoshi; Kawamoto, Toshihiro

    2003-01-01

    The estrogenic effects of bisphenol A (BPA) have been reported in human cells (E-screen assays) and in (italic)in vivo(/italic) studies of rodents, although the latter reports remain controversial, as do the exposure levels and adverse health effects of BPA in humans. In this study we report on an analytical high-performance liquid chromatography/fluorescence method for BPA and its conjugate in human urine and on the application of this method in two student cohorts. Urine, along with informa...

  11. Screening for human papillomavirus: Is urine useful?

    Directory of Open Access Journals (Sweden)

    D′Hauwers K

    2009-01-01

    Full Text Available Persistent infection with high-risk Human papillomavirus (hr-HPV 16, 18, 31, 33, and 45 is the main risk factor for developing malignant genital lesions. Screening methods and follow-up schedules for cervical cancer are well known. A golden standard to screen and monitor men does not exist yet, because HPV-related, life threatening malignancies in men are rare. The importance of male HPV screening lies mainly in HPV vaccination. Young females are the target group for HPV, but men are considered to be the reservoir for HPV and to have a role in the perpetuation of the infection in the general population. We looked at the usefulness of urine as a tool for HPV screening. Pubmed was searched with the words ′′HPV′′, ′′Urine,′′ and ′′HPV-DNA′′. The chance of finding HPV-DNA in urine is higher in men with lesions in the urethra than outside the urethra, and in women with abnormal cervical cytology. In general, the results of testing urine for HPV-DNA are better for women than for men, probably because of the anatomical position of the urethra to the vagina, vulva, and cervix. In both genders, urine HPV prevalence is higher in HIV pos patients and in high-risk populations. Urine, to screen asymptomatic low-risk-profile (women seems less useful because their urine samples are often inadequate. If urine proves to be the best medium to screen, a low-risk population remains controversial.

  12. Bisphenol A levels in human urine.

    Science.gov (United States)

    Matsumoto, Akiko; Kunugita, Naoki; Kitagawa, Kyoko; Isse, Toyohi; Oyama, Tsunehiro; Foureman, Gary L; Morita, Masatoshi; Kawamoto, Toshihiro

    2003-01-01

    The estrogenic effects of bisphenol A (BPA) have been reported in human cells (E-screen assays) and in (italic)in vivo(/italic) studies of rodents, although the latter reports remain controversial, as do the exposure levels and adverse health effects of BPA in humans. In this study we report on an analytical high-performance liquid chromatography/fluorescence method for BPA and its conjugate in human urine and on the application of this method in two student cohorts. Urine, along with information on smoking, alcohol intake, and coffee/tea consumption, was collected in two different years from two different groups of university students, 50 in 1992 and 56 in 1999. Overall, the urinary BPA levels in the students in 1992 were significantly higher than were those in 1999. The BPA levels were also positively correlated with coffee and tea consumption in the 1992 cohort but not in the 1999 cohort. We speculate that recent changes made in Japan regarding the interior coating of cans used to package these beverages may partly explain these findings.

  13. Application of electrolysis for detoxification of an antineoplastic in urine.

    Science.gov (United States)

    Kobayashi, Toyohide; Hirose, Jun; Sano, Kouichi; Kato, Ryuji; Ijiri, Yoshio; Takiuchi, Hiroya; Tanaka, Kazuhiko; Goto, Emi; Tamai, Hiroshi; Nakano, Takashi

    2012-04-01

    Antineoplastics in excreta from patients have been considered to be one of the origins of cytotoxic, carcinogenic, teratogenic, and mutagenic contaminants in surface water. Recent studies have demonstrated that antineoplastics in clinical wastewater can be detoxified by electrolysis. In this study, to develop a method for the detoxification of antineoplastics in excreta, methotrexate solution in the presence of human urine was electrolyzed and evaluated. We found that urine inhibits detoxification by electrolysis; however, this inhibition decreased by diluting urine. In urine samples, the concentrations of active chlorine generated by anodic oxidation from 0.9% NaCl solution for inactivation of antineoplastics increased in dilution-dependent and time-dependent manner. These results indicate that electrolysis with platinum-based iridium oxide composite electrode is a possible method for the detoxification of a certain antineoplastic in urine.

  14. User perceptions of urine diversion dehydration toilets: Experiences ...

    African Journals Online (AJOL)

    2013-03-20

    Mar 20, 2013 ... Experiences from a cross-sectional study in eThekwini Municipality. E Roma1,2*, K ... INTRODUCTION. Over recent years concerns have been raised within the global ..... Cultural Acceptability of using Human Excreta (Faeces and Urine) for Food ... and development of a marketing strategy. In: Werner C ...

  15. Screening for human papillomavirus: Is urine useful?

    Directory of Open Access Journals (Sweden)

    K W D′Hauwers

    2009-01-01

    We looked at the usefulness of urine as a tool for HPV screening. Pubmed was searched with the words ′′HPV′′, ′′Urine,′′ and ′′HPV-DNA′′. The chance of finding HPV-DNA in urine is higher in men with lesions in the urethra than outside the urethra, and in women with abnormal cervical cytology. In general, the results of testing urine for HPV-DNA are better for women than for men, probably because of the anatomical position of the urethra to the vagina, vulva, and cervix. In both genders, urine HPV prevalence is higher in HIV pos patients and in high-risk populations. Urine, to screen asymptomatic low-risk-profile (women seems less useful because their urine samples are often inadequate. If urine proves to be the best medium to screen, a low-risk population remains controversial.

  16. Fossilized excreta associated to dinosaurs in Brazil

    Science.gov (United States)

    Souto, P. R. F.; Fernandes, M. A.

    2015-01-01

    This work provides an updated register of the main occurrences of fossilized excreta (coprolites and urolites) associated with dinosaurs found in the Brazil. The goal is to provide a relevant guide to the interpretation of the environment in the context of Gondwana. In four geographic areas, the excreta are recovered from Cretaceous sedimentary deposits in outcrops of the Bauru and São Luis basins and the Upper Jurassic aeolian deposits of the Parana Basin in the state of São Paulo. The coprolites were analyzed by X-ray diffraction and X-ray fluorescence methods. The results of these analyses reveal compositions that differ from the surrounding matrix, indicating a partial substitution of the organic material due to the feeding habits of the producers. Additionally, we describe the urolite excavations in epirelief and hyporelief, the result of gravitational flow the impact from urine jets on sand. These are associated with ornithopod and theropod dinosaur footprints preserved in the aeolian flagstones of the Botucatu Formation, Parana Basin.

  17. Human Urine as a Noninvasive Source of Kidney Cells

    Directory of Open Access Journals (Sweden)

    Fanny Oliveira Arcolino

    2015-01-01

    Full Text Available Urine represents an unlimited source of patient-specific kidney cells that can be harvested noninvasively. Urine derived podocytes and proximal tubule cells have been used to study disease mechanisms and to screen for novel drug therapies in a variety of human kidney disorders. The urinary kidney stem/progenitor cells and extracellular vesicles, instead, might be promising for therapeutic treatments of kidney injury. The greatest advantages of urine as a source of viable cells are the easy collection and less complicated ethical issues. However, extensive characterization and in vivo studies still have to be performed before the clinical use of urine-derived kidney progenitors.

  18. Safety and Effectiveness of Struvite from Black Water and Urine as a Phosphorus Fertilizer

    NARCIS (Netherlands)

    Gell, K.; Ruijter, de F.J.; Kuntke, P.; Graaff, de M.; Smit, A.L.

    2011-01-01

    To ensure food supply, phosphorus must be recycled, for which an appealing method is using struvite fertilizer from human excreta. One struvite from black water and another from urine were assessed for safety under Dutch regulations, and for effectiveness as P fertilizer in a maize field experiment

  19. Fertiliser value of human manure from pilot urine-diversion toilets

    CSIR Research Space (South Africa)

    Mnkeni, PNS

    2009-01-01

    Full Text Available Ecological sanitation is a system that, unlike the traditional waterborne sewerage and pit toilet systems, regards human excreta as a resource to be recycled rather than as a waste. There is, however, little or no information on the fertiliser value...

  20. C26 sterol in a human urine.

    Science.gov (United States)

    Ikekawa, N; Fujimoto, Y; Isiguro, M; Suwa, S; Hirayama, Y; Mizunuma, H

    1979-06-15

    A new C26 sterol, 22-trans-27-norcholesta-5,22-dien-3 beta-ol, was found in the urine of a 6-year-old girl, with a clinical diagnosis of congenital adrenal hyperplasia of the salt losing type, accompanied by symptoms of mixed sex anatomy and skin pigmentation. The structure of the sterol was determined by comparison with the synthetic compound. The sterol was also detected in ther serum. This appears to be the first case in which a C26 sterol has occurred in mammalia.

  1. Profiles of phytoestrogens in human urine from several Asian countries.

    Science.gov (United States)

    Kunisue, Tatsuya; Tanabe, Shinsuke; Isobe, Tomohiko; Aldous, Kenneth M; Kannan, Kurunthachalam

    2010-09-08

    Intake of a diet rich in phytoestrogens has been associated with a decreased risk for hormone-dependent cancers in humans. Biomonitoring of phytoestrogens in human urine has been used to assess the intake of phytoestrogens. Although studies have reported phytoestrogen levels in urine specimens from the United States and Japan, little is known of human intake of phytoestrogens in other Asian countries. In this study we determined the concentrations of seven phytoestrogens, namely, enterolactone, enterodiol, daidzein, equol, O-desmethylangolensin (O-DMA), genistein, and coumestrol, in 199 human urine samples from three Asian countries, Vietnam (Hanoi and Ho Chi Minh), Cambodia (Phnom Penh), and India (Chennai and Kolkata), using a simple, sensitive, and reliable liquid chromatography (LC)-tandem mass spectrometry (MS/MS) method. The residue levels of phytoestrogens in urine samples from the three Asian countries were compared with the concentrations in 26 urine samples from Japan (Ehime) and 16 urine samples from the United States (Albany), analyzed in this study. Among the phytoestrogens analyzed, isoflavones such as daidzein and genistein were predominant in urine samples from Vietnam; samples from Cambodia and India contained higher concentrations of enterolactone than isoflavones. Urinary concentrations of isoflavones in samples from Hanoi, Vietnam, were notably higher than the concentrations in samples from Cambodia, India, and the United States and similar to the concentrations in samples from Japan. The lowest concentrations of daidzein and the highest concentrations of enterolactone were found in urine samples from India. Concentrations of equol and O-DMA, which are microbial transformation products of daidzein (produced by gut microflora), were notably high in urine samples from Hanoi, Vietnam. The ratios of the concentration of equol or O-DMA to that of daidzein were significantly higher in samples from Hanoi than from Japan, indicating high

  2. Impact of dietary supplemental methionine sources on sensory measurement of odor-related compounds in broiler excreta.

    Science.gov (United States)

    Chavez, C; Coufal, C D; Niemeyer, P L; Carey, J B; Lacey, R E; Miller, R K; Beier, R C

    2004-10-01

    An experiment was conducted to detect differences in odor characteristics of broiler excreta due to utilization of different supplementary Met sources by a trained human descriptive aroma attribute sensory panel. The 5 treatment groups were no supplemental Met (control group), sodium methioninate aqueous solution, dry Met hydroxy analogue, liquid Met hydroxy analogue, and DL-Met. Two trials were conducted consisting of 5 treatment groups with 3 replications of 13 randomly distributed straight run broiler chicks per pen reared in battery cages. Starter and grower diets were formulated to contain 0.5 and 0.38% Met activity, respectively (except control group, 0.35% Met activity). Excreta were collected for 24 h in litter pans lined with aluminum foil at wk 4, 5, and 6 and analyzed by a trained sensory panel (7 people). Each panelist was given 25 g of manure heated at 27 degrees C for 5 min for sensory analysis. The 13 odor attributes used to determine differences in broiler excreta by the trained sensory panel were ammonia, dirty socks, wet poultry, fermented rotten fruit, hay, musty wet, sharp, sour, urinous, rotten eggs, irritating, pungent, and nauseating. Panelist marked intensities for each attribute ranging from 0 = none and 15 = extremely intense. Each panelist was given 2 replications of each treatment group in a random order each week (total of 10 samples per wk). All data were evaluated by ANOVA using the general linear model procedure of SAS software. No significant differences were observed in BW, feed consumption, or feed conversion among the treatments. The attributes of ammonia, wet poultry, rotten fruit, musty wet, sharp, and pungent differed (P < 0.05) across treatment groups. These findings demonstrate that supplemental Met sources significantly influence odor production in broiler excreta.

  3. Bodemmaterialen voor vrijloopstallen: eigenschappen in relatie tot compostering en gasvormige emissies bij menging met mest en urine = Exploratory study of bedding materials for freestall animal housing; their composting potential and gaseous emissions after mixing with dairy cattle excreta

    NARCIS (Netherlands)

    Smits, M.C.J.; Blanken, K.; Bokma, S.; Galama, P.J.; Dooren, van H.J.C.; Szanto, G.L.

    2012-01-01

    Tussen bodemmaterialen bestaan aanzienlijke verschillen in organische stofgehalten, koolstof- en stikstofgehalten, respiratiesnelheden, temperatuurontwikkelingen en emissies bij compostering met feces en urine van melkkoeien.Chemical characteristics of bedding materials, their composting potential a

  4. Mining the human urine proteome for monitoring renal transplant injury

    Energy Technology Data Exchange (ETDEWEB)

    Sigdel, Tara K.; Gao, Yuqian; He, Jintang; Wang, Anyou; Nicora, Carrie D.; Fillmore, Thomas L.; Shi, Tujin; Webb-Robertson, Bobbie-Jo; Smith, Richard D.; Qian, Wei-Jun; Salvatierra, Oscar; Camp, David G.; Sarwal, Minnie M.

    2016-06-01

    The human urinary proteome reflects systemic and inherent renal injury perturbations and can be analyzed to harness specific biomarkers for different kidney transplant injury states. 396 unique urine samples were collected contemporaneously with an allograft biopsy from 396 unique kidney transplant recipients. Centralized, blinded histology on the graft was used to classify matched urine samples into categories of acute rejection (AR), chronic allograft nephropathy (CAN), BK virus nephritis (BKVN), and stable graft (STA). Liquid chromatography–mass spectrometry (LC-MS) based proteomics using iTRAQ based discovery (n=108) and global label-free LC-MS analyses of individual samples (n=137) for quantitative proteome assessment were used in the discovery step. Selected reaction monitoring (SRM) was applied to identify and validate minimal urine protein/peptide biomarkers to accurately segregate organ injury causation and pathology on unique urine samples (n=151). A total of 958 proteins were initially quantified by iTRAQ, 87% of which were also identified among 1574 urine proteins detected in LC-MS validation. 103 urine proteins were significantly (p<0.05) perturbed in injury and enriched for humoral immunity, complement activation, and lymphocyte trafficking. A set of 131 peptides corresponding to 78 proteins were assessed by SRM for their significance in an independent sample cohort. A minimal set of 35 peptides mapping to 33 proteins, were modeled to segregate different injury groups (AUC =93% for AR, 99% for CAN, 83% for BKVN). Urinary proteome discovery and targeted validation identified urine protein fingerprints for non-invasive differentiation of kidney transplant injuries, thus opening the door for personalized immune risk assessment and therapy.

  5. Screening for human papillomavirus: is urine useful?

    NARCIS (Netherlands)

    D'Hauwers, K.W.M.; Tjalma, W.A.

    2009-01-01

    Persistent infection with high-risk Human papillomavirus (hr-HPV 16, 18, 31, 33, and 45) is the main risk factor for developing malignant genital lesions. Screening methods and follow-up schedules for cervical cancer are well known. A golden standard to screen and monitor men does not exist yet, bec

  6. Urine Metabolite Profiles Predictive of Human Kidney Allograft Status.

    Science.gov (United States)

    Suhre, Karsten; Schwartz, Joseph E; Sharma, Vijay K; Chen, Qiuying; Lee, John R; Muthukumar, Thangamani; Dadhania, Darshana M; Ding, Ruchuang; Ikle, David N; Bridges, Nancy D; Williams, Nikki M; Kastenmüller, Gabi; Karoly, Edward D; Mohney, Robert P; Abecassis, Michael; Friedewald, John; Knechtle, Stuart J; Becker, Yolanda T; Samstein, Benjamin; Shaked, Abraham; Gross, Steven S; Suthanthiran, Manikkam

    2016-02-01

    Noninvasive diagnosis and prognostication of acute cellular rejection in the kidney allograft may help realize the full benefits of kidney transplantation. To investigate whether urine metabolites predict kidney allograft status, we determined levels of 749 metabolites in 1516 urine samples from 241 kidney graft recipients enrolled in the prospective multicenter Clinical Trials in Organ Transplantation-04 study. A metabolite signature of the ratio of 3-sialyllactose to xanthosine in biopsy specimen-matched urine supernatants best discriminated acute cellular rejection biopsy specimens from specimens without rejection. For clinical application, we developed a high-throughput mass spectrometry-based assay that enabled absolute and rapid quantification of the 3-sialyllactose-to-xanthosine ratio in urine samples. A composite signature of ratios of 3-sialyllactose to xanthosine and quinolinate to X-16397 and our previously reported urinary cell mRNA signature of 18S ribosomal RNA, CD3ε mRNA, and interferon-inducible protein-10 mRNA outperformed the metabolite signatures and the mRNA signature. The area under the receiver operating characteristics curve for the composite metabolite-mRNA signature was 0.93, and the signature was diagnostic of acute cellular rejection with a specificity of 84% and a sensitivity of 90%. The composite signature, developed using solely biopsy specimen-matched urine samples, predicted future acute cellular rejection when applied to pristine samples taken days to weeks before biopsy. We conclude that metabolite profiling of urine offers a noninvasive means of diagnosing and prognosticating acute cellular rejection in the human kidney allograft, and that the combined metabolite and mRNA signature is diagnostic and prognostic of acute cellular rejection with very high accuracy.

  7. Intercomparison of analytical methods for arsenic speciation in human urine.

    Science.gov (United States)

    Crecelius, E; Yager, J

    1997-01-01

    An intercomparison exercise was conducted for the quantification of arsenic species in spiked human urine. The primary objective of the exercise was to determine the variance among laboratories in the analysis of arsenic species such as inorganic As (As+3 and As+5), monomethylarsonic acid (MMA), and dimethylarsinic acid (DMA). Laboratories that participated had previous experience with arsenic speciation analysis. The results of this interlaboratory comparison are encouraging. There is relatively good agreement on the concentrations of these arsenic species in urine at concentrations that are relevant to research on the metabolism of arsenic in humans and other mammals. Both the accuracy and precision are relatively poor for arsenic concentrations of less than about 5 micrograms/l. PMID:9288500

  8. Natural levels of {sup 210}Po in human urine

    Energy Technology Data Exchange (ETDEWEB)

    Diaz-Frances, I.; Manjon, G.; Mantero, J.; Diaz, J. [Departament of Applied Phisic II, University of Seville, P.O. Box 41012 Seville (Spain); Garcia-Tenorio, R. [Departament of Applied Phisic II, University of Seville, P.O. Box 41012 Seville (Spain); National Accelerator Centre, P.O. Box 41092 Seville (Spain)

    2014-07-01

    Since the secret agent Alexander Litvinenko was murdered in 2006 by a {sup 210}Po lethal dose, presumably ingested, there is renovated interest on the toxicity of this radionuclide in humans. {sup 210}Po is a radioactive isotope naturally found in nature, mainly incorporated by humans via food and water ingestion, as well as inhaled through its progenitor, the {sup 222}Rn. The total amount of natural {sup 210}Po in the human body can vary from person to person depending on their lifestyle: dietary habits, drinking water source, place of residence (associated with exposure to {sup 222}Rn), etc- and therefore in the concentrations of this element to be found in urine. To analyze the influence of dietary habits on the amount of {sup 210}Po excreted in urine, two volunteers in Seville had a well-defined and time-varying diet for a month, following a daily collection of their urine and determination of the concentrations therein of this radionuclide. The results obtained and the conclusions derived from them form the core of this communication. {sup 210}Po determinations were performed daily in 200 ml aliquots of urine using the technique of high resolution alpha spectrometry. This has involved the application of a single radiochemical method for the concentration and isolation {sup 210}Po, followed by its auto-deposition on copper planchets for proper measure. Daily {sup 210}Po activity concentrations in voluntary urine analyzed during the month of study show high variability with a difference of up to an order of magnitude between maximum and minimum values obtained, and a clear dependence on the diet type followed in the various stages of the experiment. The lowest concentrations obtained are associated with a diet rich in carbohydrates and proteins 'terrestrial' (pork, beef,...), while the highest concentrations were obtained in the final phase of the experiment when the diet was enriched with presence of marine products in fair correspondence with the

  9. Human Elimination of Organochlorine Pesticides: Blood, Urine, and Sweat Study.

    Science.gov (United States)

    Genuis, Stephen J; Lane, Kevin; Birkholz, Detlef

    2016-01-01

    Background. Many individuals have been exposed to organochlorinated pesticides (OCPs) through food, water, air, dermal exposure, and/or vertical transmission. Due to enterohepatic reabsorption and affinity to adipose tissue, OCPs are not efficiently eliminated from the human body and may accrue in tissues. Many epidemiological studies demonstrate significant exposure-disease relationships suggesting OCPs can alter metabolic function and potentially lead to illness. There is limited study of interventions to facilitate OCP elimination from the human body. This study explored the efficacy of induced perspiration as a means to eliminate OCPs. Methods. Blood, urine, and sweat (BUS) were collected from 20 individuals. Analysis of 23 OCPs was performed using dual-column gas chromatography with electron-capture detectors. Results. Various OCPs and metabolites, including DDT, DDE, methoxychlor, endrin, and endosulfan sulfate, were excreted into perspiration. Generally, sweat samples showed more frequent OCP detection than serum or urine analysis. Many OCPs were not readily detected in blood testing while still being excreted and identified in sweat. No direct correlation was found among OCP concentrations in the blood, urine, or sweat compartments. Conclusions. Sweat analysis may be useful in detecting some accrued OCPs not found in regular serum testing. Induced perspiration may be a viable clinical tool for eliminating some OCPs.

  10. Human Elimination of Organochlorine Pesticides: Blood, Urine, and Sweat Study

    Directory of Open Access Journals (Sweden)

    Stephen J. Genuis

    2016-01-01

    Full Text Available Background. Many individuals have been exposed to organochlorinated pesticides (OCPs through food, water, air, dermal exposure, and/or vertical transmission. Due to enterohepatic reabsorption and affinity to adipose tissue, OCPs are not efficiently eliminated from the human body and may accrue in tissues. Many epidemiological studies demonstrate significant exposure-disease relationships suggesting OCPs can alter metabolic function and potentially lead to illness. There is limited study of interventions to facilitate OCP elimination from the human body. This study explored the efficacy of induced perspiration as a means to eliminate OCPs. Methods. Blood, urine, and sweat (BUS were collected from 20 individuals. Analysis of 23 OCPs was performed using dual-column gas chromatography with electron-capture detectors. Results. Various OCPs and metabolites, including DDT, DDE, methoxychlor, endrin, and endosulfan sulfate, were excreted into perspiration. Generally, sweat samples showed more frequent OCP detection than serum or urine analysis. Many OCPs were not readily detected in blood testing while still being excreted and identified in sweat. No direct correlation was found among OCP concentrations in the blood, urine, or sweat compartments. Conclusions. Sweat analysis may be useful in detecting some accrued OCPs not found in regular serum testing. Induced perspiration may be a viable clinical tool for eliminating some OCPs.

  11. The human urine virome in association with urinary tract infections

    Directory of Open Access Journals (Sweden)

    Tasha M Santiago-Rodriguez

    2015-01-01

    Full Text Available While once believed to represent a sterile environment, the human urinary tract harbors a unique cellular microbiota. We sought to determine whether the human urinary tract also is home to viral communities whose membership might reflect urinary tract health status. We recruited and sampled urine from 20 subjects, 10 subjects with urinary tract infections (UTIs and 10 without UTIs, and found viral communities in the urine of each subject group. Most of the identifiable viruses were bacteriophage, but eukaryotic viruses also were identified in all subjects. We found reads from human papillomaviruses (HPVs in 95% of the subjects studied, but none were found to be high-risk genotypes that are associated with cervical and rectal cancers. We verified the presence of some HPV genotypes by quantitative PCR. Some of the HPV genotypes identified were homologous to relatively novel and uncharacterized viruses that previously have been detected on skin in association with cancerous lesions, while others may be associated with anal and genital warts. On a community level, there was no association between the membership or diversity of viral communities based on urinary tract health status. While more data are still needed, detection of HPVs as members of the human urinary virome using viral metagenomics represents a non-invasive technique that could augment current screening techniques to detect low-risk HPVs in the genitourinary tracts of humans.

  12. The human urine virome in association with urinary tract infections

    Science.gov (United States)

    Santiago-Rodriguez, Tasha M.; Ly, Melissa; Bonilla, Natasha; Pride, David T.

    2014-01-01

    While once believed to represent a sterile environment, the human urinary tract harbors a unique cellular microbiota. We sought to determine whether the human urinary tract also is home to viral communities whose membership might reflect urinary tract health status. We recruited and sampled urine from 20 subjects, 10 subjects with urinary tract infections (UTIs) and 10 without UTIs, and found viral communities in the urine of each subject group. Most of the identifiable viruses were bacteriophage, but eukaryotic viruses also were identified in all subjects. We found reads from human papillomaviruses (HPVs) in 95% of the subjects studied, but none were found to be high-risk genotypes that are associated with cervical and rectal cancers. We verified the presence of some HPV genotypes by quantitative PCR. Some of the HPV genotypes identified were homologous to relatively novel and uncharacterized viruses that previously have been detected on skin in association with cancerous lesions, while others may be associated with anal and genital warts. On a community level, there was no association between the membership or diversity of viral communities based on urinary tract health status. While more data are still needed, detection of HPVs as members of the human urinary virome using viral metagenomics represents a non-invasive technique that could augment current screening techniques to detect low-risk HPVs in the genitourinary tracts of humans. PMID:25667584

  13. How do we sell the hygiene message? With dollars, dong or excreta?

    Directory of Open Access Journals (Sweden)

    West Line

    2010-06-01

    Full Text Available Abstract In North and Central Vietnam it is common among farmers to use excreta from the family double vault composting latrine (DVC as fertilizer in the fields. The official Vietnamese health guidelines stipulate a six-month period of composting before applying excreta to two of their three annual crops. However, farmers in this region cannot afford to follow these guidelines and this paper presents the reasons why. In their efforts to ensure optimal hygienic conditions, by providing a guideline, the Vietnamese health authorities have not put sufficient attention to the ‘excreta economy’ in relation to farmers’ livelihoods. The free fertilizer in the household DVC represents a value of approximately US$ 15.5 per year - or the equivalent of 15 percent of the annual household income for the poorest 20 percent of farmers. For this reason, the economic benefits derived from free fertilizer outweigh the hygiene message for most Vietnamese farmers. Even at national level the excreta economy has an impact. If Vietnam were to replace human excreta with imported fertilizer, it would involve an extra national expenditure of at least US$ 83 million a year. In order to convince Vietnamese farmers to adopt different fertilizing methods when reusing human excreta, it is necessary for the Vietnamese health authorities to change their hygiene message. They need to replace their current health sector-specific approach with a holistic one that takes the premises of farmers' livelihoods into account. If they do not the hygiene message will simply be lost.

  14. Human papillomavirus detection from human immunodeficiency virus-infected Colombian women's paired urine and cervical samples.

    Science.gov (United States)

    Munoz, Marina; Camargo, Milena; Soto-De Leon, Sara C; Sanchez, Ricardo; Parra, Diana; Pineda, Andrea C; Sussmann, Otto; Perez-Prados, Antonio; Patarroyo, Manuel E; Patarroyo, Manuel A

    2013-01-01

    Infection, coinfection and type-specific human papillomavirus (HPV) distribution was evaluated in human immunodeficiency virus (HIV)-positive women from paired cervical and urine samples. Paired cervical and urine samples (n = 204) were taken from HIV-positive women for identifying HPV-DNA presence by using polymerase chain reaction (PCR) with three generic primer sets (GP5+/6+, MY09/11 and pU1M/2R). HPV-positive samples were typed for six high-risk HPV (HR-HPV) (HPV-16, -18, -31, -33, -45 and -58) and two low-risk (LR-HPV) (HPV-6/11) types. Agreement between paired sample results and diagnostic performance was evaluated. HPV infection prevalence was 70.6% in cervical and 63.2% in urine samples. HPV-16 was the most prevalent HPV type in both types of sample (66.7% in cervical samples and 62.0% in urine) followed by HPV-31(47.2%) in cervical samples and HPV-58 (35.7%) in urine samples. There was 55.4% coinfection (infection by more than one type of HPV) in cervical samples and 40.2% in urine samples. Abnormal Papanicolau smears were observed in 25.3% of the women, presenting significant association with HPV-DNA being identified in urine samples. There was poor agreement of cervical and urine sample results in generic and type-specific detection of HPV. Urine samples provided the best diagnosis when taking cytological findings as reference. In conclusion including urine samples could be a good strategy for ensuring adherence to screening programs aimed at reducing the impact of cervical cancer, since this sample is easy to obtain and showed good diagnostic performance.

  15. Human papillomavirus detection from human immunodeficiency virus-infected Colombian women's paired urine and cervical samples.

    Directory of Open Access Journals (Sweden)

    Marina Munoz

    Full Text Available Infection, coinfection and type-specific human papillomavirus (HPV distribution was evaluated in human immunodeficiency virus (HIV-positive women from paired cervical and urine samples. Paired cervical and urine samples (n = 204 were taken from HIV-positive women for identifying HPV-DNA presence by using polymerase chain reaction (PCR with three generic primer sets (GP5+/6+, MY09/11 and pU1M/2R. HPV-positive samples were typed for six high-risk HPV (HR-HPV (HPV-16, -18, -31, -33, -45 and -58 and two low-risk (LR-HPV (HPV-6/11 types. Agreement between paired sample results and diagnostic performance was evaluated. HPV infection prevalence was 70.6% in cervical and 63.2% in urine samples. HPV-16 was the most prevalent HPV type in both types of sample (66.7% in cervical samples and 62.0% in urine followed by HPV-31(47.2% in cervical samples and HPV-58 (35.7% in urine samples. There was 55.4% coinfection (infection by more than one type of HPV in cervical samples and 40.2% in urine samples. Abnormal Papanicolau smears were observed in 25.3% of the women, presenting significant association with HPV-DNA being identified in urine samples. There was poor agreement of cervical and urine sample results in generic and type-specific detection of HPV. Urine samples provided the best diagnosis when taking cytological findings as reference. In conclusion including urine samples could be a good strategy for ensuring adherence to screening programs aimed at reducing the impact of cervical cancer, since this sample is easy to obtain and showed good diagnostic performance.

  16. Assessment Of NPK In Human Male And Female Urine For Its Fertilising Potential In Agriculture

    Directory of Open Access Journals (Sweden)

    Alfred L.K. Kuwornu

    2015-08-01

    higher in male urine than in female urine during the 2nd and 3rd months of storage. Ecosan urinals a designed urinal that seeks to separately collect urine to optimize its usefulness should be designed to separately collect urine for specific NPK requirements for crop production. Results of this study suggest that concentration of NPK in human urine is comparable to commercial chemical fertilizers. Human urine in agriculture should progressively be promoted by governments and other agencies.

  17. Analysis of human urine metabolites using SPE and NMR spectroscopy

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Nuclear magnetic resonance (NMR) spectroscopic analysis of metabonome/metabolome has widespread applications in biomedical science researches. However, most of NMR resonances for urinary metabolites remain to be fully assigned. In the present study, human urine samples from two healthy volunteers were pre-treated with C18 solid-phase extraction and the resultant 5 sub-fractions were subjected to one- and two-dimensional NMR studies, including 1H J-Resolved, 1H-1H COSY, 1H-1H TOCSY, 1H-13C HSQC, and HMBC 2D NMR. More than 70 low molecular weight metabolites were identified, and complete assignments of 1H and 13C resonances including many complex coupled spin systems were obtained.

  18. Culture of Spirulina platensis in human urine for biomass production and O2 evolution

    Institute of Scientific and Technical Information of China (English)

    Feng Dao-lun; WU Zu-cheng

    2006-01-01

    Attempts were made to culture Spirulina platensis in human urine directly to achieve biomass production and O2 evolution, for potential application to nutrient regeneration and air revitalization in life support system. The culture results showed that Spirulinaplatensis grows successfully in diluted human urine, and yields maximal biomass at urine dilution ratios of 140~240.Accumulation of lipid and decreasing of protein occurred due to N deficiency. O2 release rate of Spirulina platensis in diluted human urine was higher than that in Zarrouk medium.

  19. Dairy cow excreta patches change the boreal grass swards from sink to source of methane

    Directory of Open Access Journals (Sweden)

    Marja Elisa Maljanen

    2012-06-01

    Full Text Available We studied methane (CH4 flux rates from experimental excreta patches on a dairy pasture with a chamber technique during snow free seasons and with a gas gradient technique during winter from timothy-meadow fescue sward with mineral N fertilization (220 kg ha-1 and from grass-white clover mixture without fertilization. The dung and urine patches were applied in June or August two consecutive grazing seasons and the measurements were carried out for a year following each application. There were no significant differences in CH4 fluxes between plant species and emissions originated mainly from the fresh dung pats. The average annual CH4 fluxes from the control sites without excreta were -0.60±0.1 and with the excreta 0.47±0.3 kg CH4 ha-1. Thus, excreta originating from dairy cows can turn boreal swards from weak sinks to small sources of CH4. However, these emissions are only 0.2% of the total CH4 emissions from a dairy cow.

  20. Identification of Metabolites of the Fungicide Penconazole in Human Urine.

    Science.gov (United States)

    Mercadante, R; Polledri, E; Scurati, S; Moretto, A; Fustinoni, S

    2016-07-18

    Penconazole (PEN) is a fungicide used in agriculture that has been classified as hazardous to humans and the environment. The objective of this work was to identify PEN urinary metabolites in humans and propose a biomarker for PEN exposure. Five urine samples were collected from agricultural workers who worked with and were exposed to PEN. Samples were analyzed by liquid chromatography coupled with hybrid triple quadrupole-linear ion trap mass spectrometry, with the source operating in the electrospray ionization mode. Metabolites previously identified in animal studies were searched as possible metabolites in humans. Candidate metabolites were first identified by multiple reaction monitoring following the protonated molecular ions that generated the protonated triazole moiety, which is expected to be present in all PEN metabolites; second, the isotopic patterns of the molecular ions were checked for consistency with the presence of two chlorine atoms; third, the full mass spectra were evaluated for consistency with the molecular structure. Seven different oxidized metabolites were found, both in the free and glucuronide conjugate forms. The major metabolite was the monohydroxyl-derivative PEN-OH (median molar fraction approximately 0.92 as a sum of free and glucuronide conjugated form). The product of further oxidation was the carboxyl-derivate PEN-COOH (median molar fraction approximately 0.03). After hydrolysis with β-glucuronidase, the free compounds were quantified in the presence of deuterated PEN as an internal standard; PEN-OH levels ranged from 230 to 460 μg/L, and PEN-COOH levels ranged from 5.2 to 16.7 μg/L. We propose a pathway for PEN metabolism in humans and suggest PEN-OH, after hydrolysis of glucuronide conjugates, as a biomarker for monitoring human exposure to PEN.

  1. How do we sell the hygiene message? With dollars, dong or excreta?

    DEFF Research Database (Denmark)

    Jensen, Peter Kjaer Mackie; Phuc, Pham Duc; West, Line Gram Knudsen

    2010-01-01

    different fertilizing methods when reusing human excreta, it is necessary for the Vietnamese health authorities to change their hygiene message. They need to replace their current health sector-specific approach with a holistic one that takes the premises of farmers' livelihoods into account. If they do...... to farmers' livelihoods. The free fertilizer in the household DVC represents a value of approximately US$ 15.5 per year--or the equivalent of 15 percent of the annual household income for the poorest 20 percent of farmers. For this reason, the economic benefits derived from free fertilizer outweigh...... the hygiene message for most Vietnamese farmers. Even at national level the excreta economy has an impact. If Vietnam were to replace human excreta with imported fertilizer, it would involve an extra national expenditure of at least US$ 83 million a year.In order to convince Vietnamese farmers to adopt...

  2. Identification of tetrahydro-beta-carboline-3-carboxylic acid in foodstuffs, human urine and human milk.

    Science.gov (United States)

    Adachi, J; Mizoi, Y; Naito, T; Ogawa, Y; Uetani, Y; Ninomiya, I

    1991-05-01

    1-Methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (MTCA) and 1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (TCCA), both precursors of mutagenic N-nitroso compounds (N-nitrosamines, 1-methyl-2-nitroso-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid and 2-nitroso-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid), were detected in various food-stuffs, urine from healthy human subjects and human milk. A purification procedure, involving a chemically-bonded material followed by HPLC combined with fluorometric detection, was used for the quantitative determination of these compounds, allowing the separation of two diastereoisomers of MTCA. An HPLC and mass spectrometry method was also developed for their identification. Comparing the concentration of MTCA and TCCA in fermented products and raw materials suggested that tetrahydro-beta-carbolines may have been produced through fermentation or by condensation of tryptophan and acetaldehyde formed from ethanol added as a food preservative. This is the first report of excretion of tetrahydro-beta-carbolines in human urine and human milk. A comparison of the concentrations of tetrahydro-beta-carbolines in urine from human infants and human milk indicates that tetrahydro-beta-carbolines may be synthesized endogenously in humans. A possible pathway of tryptophan metabolism in plants and animals is presented.

  3. Urine versus brushed samples in human papillomavirus screening: study in both genders.

    NARCIS (Netherlands)

    Hauwers, K.W.M. d'; Depuydt, C.; Bogers, J.P.; Stalpaert, M.; Vereecken, A.; Wyndaele, J.J.; Tjalma, W.

    2007-01-01

    AIM: To investigate whether urine is a good medium for screening and whether there is a correlation between the amount of extracted DNA and human papillomavirus (HPV)-positivity. METHODS: In the present study, 30 first-voided urine (FVU) specimens and 20 urethroglandular swabs using cervex-brushes f

  4. Global gene expression profiling of asymptomatic bacteriuria Escherichia coli during biofilm growth in human urine

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Klemm, Per

    2007-01-01

    asymptomatic bacteriuria (ABU) E. coli strains 83972 and VR50 are significantly better biofilm formers in their natural growth medium, human urine, than the two uropathogenic E. coli isolates CFT073 and 536. We used DNA microarrays to monitor the expression profile during biofilm growth in urine of the two ABU...

  5. Urine antigen detection for the diagnosis of human neurocysticercosis.

    Science.gov (United States)

    Castillo, Yesenia; Rodriguez, Silvia; García, Hector H; Brandt, Jef; Van Hul, Anke; Silva, Maria; Rodriguez-Hidalgo, Richar; Portocarrero, Mylagritos; Melendez, D Paolo; Gonzalez, Armando E; Gilman, Robert H; Dorny, Pierre

    2009-03-01

    Neurocysticercosis (NCC) is a major cause of seizures and epilepsy. Diagnosis is based on brain imaging, supported by immunodiagnosis in serum or cerebrospinal fluid (CSF). Lumbar puncture is invasive and painful. Blood sampling is slightly painful and poorly accepted. Urine antigen detection has been used for other parasites and tried in NCC with suboptimal performance. We used a monoclonal antibody-based ELISA to detect Taenia solium antigens in urine from 87 Peruvian neurocysticercosis patients (viable cysts, N = 34; subarachnoid cysticercosis, N = 10; degenerating parasites, N = 7; calcified lesions, N = 36) and 32 volunteers from a non-endemic area of Peru. Overall sensitivity of urine antigen detection for viable parasites was 92%, which decreased to 62.5% in patients with a single cyst. Most patients (30/36, 83%) with only calcified cysticercosis were urine antigen negative. Antigen levels in paired serum/urine samples (evaluated in 19 patients) were strongly correlated. Non-invasive urine testing for T. solium antigens provides a useful alternative for NCC diagnosis.

  6. Identification of factors in human urine that inhibit the binding of Escherichia coli adhesins.

    OpenAIRE

    1988-01-01

    Earlier studies on the binding of Escherichia coli adhesins to the human urinary tract have indicated that the ability to recognize binding sites on the urinary tract epithelial cells is not a characteristic for P fimbriae only, but is also shared by some other adhesins that are not associated with pyelonephritis, especially S fimbriae. In the present study we have investigated whether human urine contains inhibitors of the binding of E. coli adhesins. Normal human urine was found to inhibit ...

  7. Effects of N source and nitrification pretreatment on growth of Arthrospira platensis in human urine

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Culture of Arthrospira platensis (Spirulina platensis) in human urine was investigated to get valuable biomass. NO3-N was the proper N source, in comparison with other N source, includ ing urea, NH4-N and NO2-N. As a result, aerobic nitrification of human urine was performed, with above 93.6% nitrification percentage finally achieved with total-N (TN) load of 46.52those in Zarrouk medium. Thus, it is possible to culture Arthrospiraplatensis in nitrified human urine for food production within bioregenerative life support systems (BLSSs).

  8. Economic and nutrient discharge tradeoffs of excreta-fed aquaculture in the Mekong Delta, Vietnam

    NARCIS (Netherlands)

    Nhan, D.K.; Verdegem, M.C.J.; Binh, N.T.; Duong, L.T.; Milstein, A.; Verreth, J.A.J.

    2008-01-01

    The present study quantifies the effects on production, nutrient discharge and economic return of the use of pig and human excreta in pond tanning. On-farm data from various studies were integrated and analyzed applying single and multiple regression methods. Pond-dissolved oxygen concentration, wat

  9. Monitoring Homovanillic Acid and Vanillylmandelic Acid in Human Urine by Capillary Electrophoresis with Electrochemical Detection

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    A simple, rapid and low-cost method of separation and determination of homovanillic acid and vanillylmandelic acid in human urine was developed based on capillary zone electrophoresis / amperometric detection with high sensitivity and good resolution.

  10. Urea hydrolysis and recovery of nitrogen and phosphorous as MAP from stale human urine

    Institute of Scientific and Technical Information of China (English)

    LIU Zhigang; ZHAO Qingliang; WANG Kun; LEE Duujong; QIU Wei; WANG Jianfang

    2008-01-01

    Laboratory-scale tests for magnesium ammonium phosphate (MAP) precipitation following urea hydrolysis of human urine were conducted using orthogonal experiment design. The effects of initial pH, temperature and the volumetric ratios of stale urine to fresh urine, on urea hydrolysis in urine were studied to determine the final hydrolysis time to recover most nitrogen from separated human urine by MAP. With a volumetric ratio of stale to fresh urine >10% and at temperature of 20℃ and above, urea hydrolysis could be completed in two days. Alkaline pH inhibited urea hydrolysis progress. The final pHs were all around 9.0 following urine hydrolysis, while the suspension pH might act as an indicator to detect the start and extent of urea hydrolysis. Over 95% of ammonium nitrogen and over 85% of phosphorus from hydrolyzed urine as MAP precipitate were obtained using MgCl2·6H2O and Na2HPO4·12H2O as precipitation agents at pH 8.5, molar ratio of Mg2+:NH4+-N:PO43--P at (1.2--1.3):1:1, mixing speed of 120 r/min, and precipitation time and reaction time of 3 h and 15 min, respectively. The precipitate has a structure resembling pure MAP crystal.

  11. Global gene expression profiling of asymptomatic bacteriuria Escherichia coli during biofilm growth in human urine

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Klemm, Per

    2007-01-01

    asymptomatic bacteriuria (ABU) E. coli strains 83972 and VR50 are significantly better biofilm formers in their natural growth medium, human urine, than the two uropathogenic E. coli isolates CFT073 and 536. We used DNA microarrays to monitor the expression profile during biofilm growth in urine of the two ABU...... strains 83972 and VR50. Significant differences in expression levels were seen between the biofilm expression profiles of the two strains with the corresponding planktonic expression profiles in morpholinepropanesulfonic acid minimal laboratory medium and human urine; 417 and 355 genes were up- and down...... versions of 83972 and VR50; all mutants showed reduced biofilm formation in urine by 18 to 43% compared with the wild type (P profile of strain 83972 in the human urinary tract partially overlaps with the biofilm expression profile....

  12. Discovery and Characterization of Human-Urine Utilization by Asymptomatic-Bacteriuria-Causing Streptococcus agalactiae.

    Science.gov (United States)

    Ipe, Deepak S; Ben Zakour, Nouri L; Sullivan, Matthew J; Beatson, Scott A; Ulett, Kimberly B; Benjamin, William H; Davies, Mark R; Dando, Samantha J; King, Nathan P; Cripps, Allan W; Schembri, Mark A; Dougan, Gordon; Ulett, Glen C

    2015-11-09

    Streptococcus agalactiae causes both symptomatic cystitis and asymptomatic bacteriuria (ABU); however, growth characteristics of S. agalactiae in human urine have not previously been reported. Here, we describe a phenotype of robust growth in human urine observed in ABU-causing S. agalactiae (ABSA) that was not seen among uropathogenic S. agalactiae (UPSA) strains isolated from patients with acute cystitis. In direct competition assays using pooled human urine inoculated with equal numbers of a prototype ABSA strain, designated ABSA 1014, and any one of several UPSA strains, measurement of the percentage of each strain recovered over time showed a markedly superior fitness of ABSA 1014 for urine growth. Comparative phenotype profiling of ABSA 1014 and UPSA strain 807, isolated from a patient with acute cystitis, using metabolic arrays of >2,500 substrates and conditions revealed unique and specific l-malic acid catabolism in ABSA 1014 that was absent in UPSA 807. Whole-genome sequencing also revealed divergence in malic enzyme-encoding genes between the strains predicted to impact the activity of the malate metabolic pathway. Comparative growth assays in urine comparing wild-type ABSA and gene-deficient mutants that were functionally inactivated for the malic enzyme metabolic pathway by targeted disruption of the maeE or maeK gene in ABSA demonstrated attenuated growth of the mutants in normal human urine as well as synthetic human urine containing malic acid. We conclude that some S. agalactiae strains can grow in human urine, and this relates in part to malic acid metabolism, which may affect the persistence or progression of S. agalactiae ABU.

  13. Energy efficient reconcentration of diluted human urine using ion exchange membranes in bioelectrochemical systems.

    Science.gov (United States)

    Tice, Ryan C; Kim, Younggy

    2014-11-01

    Nutrients can be recovered from source separated human urine; however, nutrient reconcentration (i.e., volume reduction of collected urine) requires energy-intensive treatment processes, making it practically difficult to utilize human urine. In this study, energy-efficient nutrient reconcentration was demonstrated using ion exchange membranes (IEMs) in a microbial electrolysis cell (MEC) where substrate oxidation at the MEC anode provides energy for the separation of nutrient ions (e.g., NH4(+), HPO4(2-)). The rate of nutrient separation was magnified with increasing number of IEM pairs and electric voltage application (Eap). Ammonia and phosphate were reconcentrated from diluted human urine by a factor of up to 4.5 and 3.0, respectively (Eap = 1.2 V; 3-IEM pairs). The concentrating factor increased with increasing degrees of volume reduction, but it remained stationary when the volume ratio between the diluate (urine solution that is diluted in the IEM stack) and concentrate (urine solution that is reconcentrated) was 6 or greater. The energy requirement normalized by the mass of nutrient reconcentrated was 6.48 MJ/kg-N (1.80 kWh/kg-N) and 117.6 MJ/kg-P (32.7 kWh/kg-P). In addition to nutrient separation, the examined MEC reactor with three IEM pairs showed 54% removal of COD (chemical oxygen demand) in 47-hr batch operation. The high sulfate concentration in human urine resulted in substantial growth of both of acetate-oxidizing and H2-oxidizing sulfate reducing bacteria, greatly diminishing the energy recovery and Coulombic efficiency. However, the high microbial activity of sulfate reducing bacteria hardly affected the rate of nutrient reconcentration. With the capability to reconcentrate nutrients at a minimal energy consumption and simultaneous COD removal, the examined bioelectrochemical treatment method with an IEM application has a potential for practical nutrient recovery and sustainable treatment of source-separated human urine.

  14. Serum and Urine Biomarkers for Human Renal Cell Carcinoma

    Directory of Open Access Journals (Sweden)

    A. L. Pastore

    2015-01-01

    Full Text Available Renal cell carcinoma (RCC diagnosis is mostly achieved incidentally by imaging provided for unrelated clinical reasons. The surgical management of localized tumors has reported excellent results. The therapy of advanced RCC has evolved considerably over recent years with the widespread use of the so-called “targeted therapies.” The identification of molecular markers in body fluids (e.g., sera and urine, which can be used for screening, diagnosis, follow-up, and monitoring of drug-based therapy in RCC patients, is one of the most ambitious challenges in oncologic research. Although there are some promising reports about potential biomarkers in sera, there is limited available data regarding urine markers for RCC. The following review reports some of the most promising biomarkers identified in the biological fluids of RCC patients.

  15. Determination of ortho-phenylphenol in human urine by gas chromatography-mass spectrometry.

    Science.gov (United States)

    Bartels, M J; Brzak, K A; Bormett, G A

    1997-12-05

    A sensitive gas chromatographic-mass spectrometric method was developed to quantitate total o-phenylphenol (OPP) (free plus conjugates) in human urine. Conjugates of OPP were acid-hydrolyzed to free OPP, derivatized to the pentafluorobenzoyl ester derivative and analyzed via negative-ion chemical ionization gas chromatography-mass spectrometry. Two stable isotope analogs of OPP were shown to be suitable as internal standards for this method (D2-phenol ring, 13C6-phenyl ring). A synthetic method is presented for the preparation of the D2-OPP internal standard. The 13C6-OPP analog was also shown to be useful as an alternate test material for laboratory-based exposure studies. The limit of quantitation for this method was 1 ng OPP/ml urine. Calibration curves were linear for the analyte over the concentration range of 0.5-1117 ng OPP/ml urine. Relative recovery of OPP from urine ranged from 97.0 to 104.7%. Low levels of OPP (mean=6+/-7 ng/ml; n=22) were found in control human urine samples. The method was validated with urine samples obtained from human volunteers undergoing a dermal exposure study with 12C-/13C6-/14C-OPP. This method was developed to aid in assessments of human exposure to OPP during a variety of uses of the compound.

  16. Genotyping of Pseudomonas aeruginosa isolated from cockroaches and human urine.

    Science.gov (United States)

    Saitou, Keiko; Furuhata, Katsunori; Fukuyama, Masafumi

    2010-10-01

    Molecular-epidemiological analysis of Pseudomonas aeruginosa isolated from cockroaches captured in hospitals and from patient urine was performed, employing randomly amplified polymorphic DNA (RAPD) analysis to investigate the usefulness of RAPD analysis. Four specific bands at positions of 993, 875, 521, and 402 bp were commonly detected using primer 272 in 16 of 45 cockroach-derived strains (35.6%), but not in 21 urine-derived strains. On analysis using primer 208, 4 specific bands at positions of 1,235, 1,138, 1,068, and 303 bp were commonly detected in 15 of the 45 cockroach-derived (33.3%) and 10 of the 21 patient urine-derived (47.6%) strains, in a total of 25 of 66 strains (37.8%). On cluster analysis, 12 (48.5%) and 16 (66.7%) clusters were grouped based on a homology of 89% or greater, using primer 272 and primer 208, respectively, showing that primer 208 was suitable for the confirmation of diversity. Seven patterns were clustered based on 100% homology using either primer, and 6 of these consisted of only cockroach-derived strains. In the individual groups with 100% homology, all strains in the group were isolated at an identical site during the same period. P. aeruginosa isolated from cockroaches showed diverse genotypes suggesting several sources of contamination, indicating the necessity for investigating infection control targeting cockroaches inhabiting hospitals.

  17. Sheep Excreta as Source of Nitrous Oxide in Ryegrass Pasture in Southern Brazil

    Directory of Open Access Journals (Sweden)

    Michely Tomazi

    2015-10-01

    Full Text Available ABSTRACT Livestock urine and dung are important components of the N cycle in pastures, but little information on its effect on soil nitrous oxide (N2O emissions is available. We conducted a short-term (39-day trial to quantify the direct N2O-N emissions from sheep excreta on an experimental area of ryegrass pasture growing on a Typic Paleudult in southern Brazil. Four rates of urine-N (161, 242, 323, and 403 kg ha-1 N and one of dung-N (13 kg ha-1 N were applied, as well as a control plot receiving no excreta. The N2O-N emission factor (EF = % of added N released as N2O-N for urine and dung was calculated, taking into account the N2O fluxes in the field, over a period of 39 days. The EF value of the urine and dung was used to estimate the emissions of N2O-N over a 90-day period of pasture in the winter under two grazing intensities (2.5 or 5.0 times the herbage intake potential of grazing lambs. The soil N2O-N fluxes ranged from 4 to 353 µg m-2h-1. The highest N2O-N fluxes occurred 16 days after application of urine and dung, when the highest soil nitrate content was also recorded and the water-filled pore space exceeded 60 %. The mean EF for urine was 0.25 % of applied N, much higher than that for dung (0.06 %. We found that N2O-N emissions for the 90-day winter pasture period were 0.54 kg ha-1 for low grazing intensity and 0.62 kg ha-1 for moderate grazing intensity. Comparison of the two forms of excreta show that urine was the main contributor to N2O-N emissions (mean of 36 %, whereas dung was responsible for less than 0.1 % of total soil N2O-N emissions.

  18. 20 CFR 654.406 - Excreta and liquid waste disposal.

    Science.gov (United States)

    2010-04-01

    ... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Excreta and liquid waste disposal. 654.406 Section 654.406 Employees' Benefits EMPLOYMENT AND TRAINING ADMINISTRATION, DEPARTMENT OF LABOR SPECIAL RESPONSIBILITIES OF THE EMPLOYMENT SERVICE SYSTEM Housing for Agricultural Workers Housing Standards § 654.406 Excreta and liquid waste disposal....

  19. Toilet compost and human urine used in agriculture: fertilizer value assessment and effect on cultivated soil properties.

    Science.gov (United States)

    Sangare, D; Sou Dakoure, M; Hijikata, N; Lahmar, R; Yacouba, H; Coulibaly, L; Funamizu, N

    2015-01-01

    Toilet compost (TC) and human urine are among natural fertilizers, which raise interest due to their double advantages to combine sanitation and nutrient recovery. However, combination of urine and TC is not so spread probably because the best ratio (urine/TC) is still an issue and urine effect on soil chemical properties remains poorly documented. This study aims to determine the best ratio of urine and TC in okra cultivation, by targeting higher fertilization effect combined with lower impact on soil chemical properties. Based on Nitrogen requirement of okra, seven treatments were compared: (T0) no fertilizer, (T1) chemical fertilizer (NPK: 14-23-14), (T2) 100% urine, (T3) 100% TC, (T4) ratio of 75% urine+25% TC, (T5) 50% urine+50% TC and (T6) 25% urine+75% TC. Results indicated that T4 (75% urine+25% TC) gave the highest plant height and yield. In contrast, T2 (100% urine) gave the lowest results among all treatments, indicating toxicity effects on plant growth and associated final yield. Such toxicity is confirmed by soil chemical properties at T2 with soil acidification and significant increase in soil salinity. In contrast, application of urine together with TC mitigates soil acidification and salinity, highlighting the efficiency of urine and TC combination on soil chemical properties. However, further investigation is necessary to refine better urine/TC ratio for okra production.

  20. Utilization of Human Urine as Fertilizer with Magnesium Oxide (MgO, Zeolite and Activated Carbon as Absorbent

    Directory of Open Access Journals (Sweden)

    Hijrah Purnama Putra

    2014-01-01

    Full Text Available Urine is residual fluid excreted by kidneys through urinary tract to outside of the human body, to maintain homeostasis of fluid in the body. Normally urine still contains high amount of nitrogen, which is 87%, phosphor 50%, potassium 54% and low bacterial content. With these contents urine potentially becomes organic fertilizer rich with nitrogen, phosphor and potassium contents and is beneficial to plants. However, until today the utilization or urine in Indonesia is very low. The urine produced is dispose with feces in toilets. This study aimed to utilize urine as solid organic fertilizer using magnesium oxide (MgO, zeolite, and actived carbon as absorbents of ammonium and phosphor. The study started with collecting urine, time variations of urine storage were 24; 48 and 72 hours, and urine was mixed with water as an assumption that urine mixes with water when flushed in urinals. The result showed effectiveness of optimum urine absorption in urine stored for 48 hours by adding 8 gram MgO, producing ammonium and phosphor contents 56.100 ppm and 3.610 ppm, respectively. From environmental perspective, utilization of urine as organic fertilizer was applicable because it satisfied the ecological principle of sanitation to prevent soil pollution, ground and surface water pollution and its utilization as agricultural resources.

  1. Determination of monomethylarsonous acid, a key arsenic methylation intermediate, in human urine.

    Science.gov (United States)

    Le, X C; Ma, M; Cullen, W R; Aposhian, H V; Lu, X; Zheng, B

    2000-01-01

    In this study we report on the finding of monomethylarsonous acid [MMA(III)] in human urine. This newly identified arsenic species is a key intermediate in the metabolic pathway of arsenic biomethylation, which involves stepwise reduction of pentavalent to trivalent arsenic species followed by oxidative addition of a methyl group. Arsenic speciation was carried out using ion-pair chromatographic separation of arsenic compounds with hydride generation atomic fluorescence spectrometry detection. Speciation of the inorganic arsenite [As(III)], inorganic arsenate [As(V)], monomethylarsonic acid [MMA(V)], dimethylarsinic acid [DMA(V)], and MMA(III) in a urine sample was complete in 5 min. Urine samples collected from humans before and after a single oral administration of 300 mg sodium 2,3-dimercapto-1-propane sulfonate (DMPS) were analyzed for arsenic species. MMA(III) was found in 51 out of 123 urine samples collected from 41 people in inner Mongolia 0-6 hr after the administration of DMPS. MMA(III )in urine samples did not arise from the reduction of MMA(V) by DMPS. DMPS probably assisted the release of MMA(III) that was formed in the body. Along with the presence of MMA(III), there was an increase in the relative concentration of MMA(V) and a decrease in DMA(V) in the urine samples collected after the DMPS ingestion. PMID:11102289

  2. Identification of factors in human urine that inhibit the binding of Escherichia coli adhesins.

    Science.gov (United States)

    Parkkinen, J; Virkola, R; Korhonen, T K

    1988-10-01

    Earlier studies on the binding of Escherichia coli adhesins to the human urinary tract have indicated that the ability to recognize binding sites on the urinary tract epithelial cells is not a characteristic for P fimbriae only, but is also shared by some other adhesins that are not associated with pyelonephritis, especially S fimbriae. In the present study we have investigated whether human urine contains inhibitors of the binding of E. coli adhesins. Normal human urine was found to inhibit hemagglutination by S and type 1 fimbriae but not P fimbriae. The major inhibitor of S fimbriae in normal urine was identified as Tamm-Horsfall glycoprotein, and the interaction with S fimbriae is probably mediated by its sialyloligosaccharide chains. No significant variation was observed in the inhibitory effect of T-H glycoprotein preparations originating from different individuals. In contrast to S fimbriae, the major inhibitors of type 1 fimbriae in urine were identified as low-molecular-weight compounds. Gel filtration and ion-exchange chromatography and alpha-mannosidase treatment indicated that they were neutral alpha-mannosides, probably manno-oligosaccharides with three to five saccharides. Studies of urine samples collected from several individuals indicated the common occurrence of these inhibitory alpha-mannosides. Type 1 fimbriae bound to immobilized T-H glycoprotein, but, unlike S fimbriae, their binding was poorly inhibited by soluble T-H glycoprotein. Some urine samples were also found to contain low-molecular-weight inhibitors for the O75X adhesin of E. coli. These results emphasize that to function as a virulence factor in human urinary tract infections, an adhesin must evidently recognize such receptor structures at the infection sites that are not excreted in soluble form in urine. This prerequisite is filled by P fimbriae but not by type 1 or S fimbriae.

  3. Analysis of endogenous aldehydes in human urine by static headspace gas chromatography-mass spectrometry.

    Science.gov (United States)

    Serrano, María; Gallego, Mercedes; Silva, Manuel

    2016-03-11

    Endogenous aldehydes (EAs) generated during oxidative stress and cell processes are associated with many pathogenic and toxicogenic processes. The aim of this research was to develop a solvent-free and automated analytical method for the determination of EAs in human urine using a static headspace generator sampler coupled with gas chromatography-mass spectrometry (HS-GC-MS). Twelve significant EAs used as markers of different biochemical and physiological processes, namely short- and medium-chain alkanals, α,β-unsaturated aldehydes and dicarbonyl aldehydes have been selected as target analytes. Human urine samples (no dilution is required) were derivatized with O-2,3,4,5,6-pentafluorobenzylhydroxylamine in alkaline medium (hydrogen carbonate-carbonate buffer, pH 10.3). The HS-GC-MS method developed renders an efficient tool for the sensitive and precise determination of EAs in human urine with limits of detection from 1 to 15ng/L and relative standard deviations, (RSDs) from 6.0 to 7.9%. Average recoveries by enriching urine samples ranged between 92 and 95%. Aldehydes were readily determined at 0.005-50μg/L levels in human urine from healthy subjects, smokers and diabetic adults.

  4. Integrated forward osmosis-membrane distillation process for human urine treatment.

    Science.gov (United States)

    Liu, Qianliang; Liu, Caihong; Zhao, Lei; Ma, Weichao; Liu, Huiling; Ma, Jun

    2016-03-15

    This study demonstrated a forward osmosis-membrane distillation (FO-MD) hybrid system for real human urine treatment. A series of NaCl solutions at different concentrations were adopted for draw solutions in FO process, which were also the feed solutions of MD process. To establish a stable and continuous integrated FO-MD system, individual FO process with different NaCl concentrations and individual direct contact membrane distillation (DCMD) process with different feed temperatures were firstly investigated separately. Four stable equilibrium conditions were obtained from matching the water transfer rates of individual FO and MD processes. It was found that the integrated system is stable and sustainable when the water transfer rate of FO subsystem is equal to that of MD subsystem. The rejections to main contaminants in human urine were also investigated. Although individual FO process had relatively high rejection to Total Organic Carbon (TOC), Total Nitrogen (TN) and Ammonium Nitrogen (NH4(+)-N) in human urine, these contaminants could also accumulate in draw solution after long term performance. The MD process provided an effective rejection to contaminants in draw solution after FO process and the integrated system revealed nearly complete rejection to TOC, TN and NH4(+)-N. This work provided a potential treatment process for human urine in some fields such as water regeneration in space station and water or nutrient recovery from source-separated urine.

  5. NMR metabolomics of human blood and urine in disease research.

    Science.gov (United States)

    Duarte, Iola F; Diaz, Sílvia O; Gil, Ana M

    2014-05-01

    This paper reviews the main applications of NMR metabolomics of blood and urine in disease research, over the last 5 years. The broad range of disease types addressed attests the increasing interest within the academic and medical communities to explore the recognised potential of metabolomics to (1) provide insight into underlying disease pathogenesis and (2) unveil new metabolic markers for disease diagnosis and follow up. Importantly, most recent studies reveal an increasing awareness of possible limitations and pitfalls of the metabolomics approach, together with efforts for improved study design and statistical validation, which are crucial requisites for the sound development of NMR metabolomics and its progress into the clinical setting. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. Quantitative Determination of Common Urinary Odorants and Their Glucuronide Conjugates in Human Urine

    Directory of Open Access Journals (Sweden)

    Maria Wagenstaller

    2013-08-01

    Full Text Available Our previous study on the identification of common odorants and their conjugates in human urine demonstrated that this substance fraction is a little-understood but nonetheless a promising medium for analysis and diagnostics in this easily accessible physiological medium. Smell as an indicator for diseases, or volatile excretion in the course of dietary processes bares high potential for a series of physiological insights. Still, little is known today about the quantitative composition of odorous or volatile targets, as well as their non-volatile conjugates, both with regard to their common occurrence in urine of healthy subjects, as well as in that of individuals suffering from diseases or other physiological misbalancing. Accordingly, the aim of our study was to develop a highly sensitive and selective approach to determine the common quantitative composition of selected odorant markers in healthy human subjects, as well as their corresponding glucuronide conjugates. We used one- and two-dimensional high resolution gas chromatography-mass spectrometry in combination with stable isotope dilution assays to quantify commonly occurring and potent odorants in human urine. The studies were carried out on both native urine and on urine that had been treated by glucuronidase assays, with analysis of the liberated odor-active compounds using the same techniques. Analytical data are discussed with regard to their potential translation as future diagnostic tool.

  7. Human Excretion of Bisphenol A: Blood, Urine, and Sweat (BUS Study

    Directory of Open Access Journals (Sweden)

    Stephen J. Genuis

    2012-01-01

    Full Text Available Background. Bisphenol A (BPA is an ubiquitous chemical contaminant that has recently been associated with adverse effects on human health. There is incomplete understanding of BPA toxicokinetics, and there are no established interventions to eliminate this compound from the human body. Using 20 study participants, this study was designed to assess the relative concentration of BPA in three body fluids—blood, urine, and sweat—and to determine whether induced sweating may be a therapeutic intervention with potential to facilitate elimination of this compound. Methods. Blood, urine, and sweat were collected from 20 individuals (10 healthy participants and 10 participants with assorted health problems and analyzed for various environmental toxicants including BPA. Results. BPA was found to differing degrees in each of blood, urine, and sweat. In 16 of 20 participants, BPA was identified in sweat, even in some individuals with no BPA detected in their serum or urine samples. Conclusions. Biomonitoring of BPA through blood and/or urine testing may underestimate the total body burden of this potential toxicant. Sweat analysis should be considered as an additional method for monitoring bioaccumulation of BPA in humans. Induced sweating appears to be a potential method for elimination of BPA.

  8. Solar thermal evaporation of human urine for nitrogen and phosphorus recovery in Vietnam

    Energy Technology Data Exchange (ETDEWEB)

    Antonini, Samantha, E-mail: sam_antonini@uni-bonn.de; Nguyen, Phong Thanh; Arnold, Ute; Eichert, Thomas; Clemens, Joachim

    2012-01-01

    A No Mix sanitation system was installed in a dormitory at the University of Can Tho in Vietnam, with the objective of recycling nutrients from source separated urine. This paper presents a pilot scale evaporation technology, and investigates the feasibility of recovering nitrogen and phosphorus from human urine by solar still for use as fertilizer. After 26 days of sun exposure, 360 g of solid fertilizer material was recovered from 50 L undiluted urine. This urine-derived fertilizer was mainly composed of sodium chloride, and had phosphorus and nitrogen contents of almost 2%. When tested with maize and ryegrass, the urine fertilizer led to biomass yields and phosphorus and nitrogen uptakes comparable to those induced by a commercial mineral fertilizer. Urine acidification with sulfuric or phosphoric acid prior treatment reduced nitrogen losses, improved the nutrient content of the generated fertilizers, and induced higher biomass yields and nitrogen and phosphorus uptakes than the commercial mineral fertilizer. However, acidification is not recommended in developing countries due to additional costs and handling risks. The fate of micropollutants and the possibility of separating sodium chloride from other beneficial nutrients require further investigation. - Highlights: Black-Right-Pointing-Pointer 360 g of fertilizer was derived from 50 L urine by solar evaporative distillation. Black-Right-Pointing-Pointer The fertilizer contained 90% sodium chloride, 3% sulfur, 2% nitrogen, 2% phosphorus. Black-Right-Pointing-Pointer It induced biomass yields comparable to those produced by a commercial fertilizer. Black-Right-Pointing-Pointer Urine acidification improved the nutrient content of the generated fertilizers. Black-Right-Pointing-Pointer Acidification is not recommended for use in developing countries (costs, safety).

  9. A magnetic bead-based method for concentrating DNA from human urine for downstream detection.

    Science.gov (United States)

    Bordelon, Hali; Russ, Patricia K; Wright, David W; Haselton, Frederick R

    2013-01-01

    Due to the presence of PCR inhibitors, PCR cannot be used directly on most clinical samples, including human urine, without pre-treatment. A magnetic bead-based strategy is one potential method to collect biomarkers from urine samples and separate the biomarkers from PCR inhibitors. In this report, a 1 mL urine sample was mixed within the bulb of a transfer pipette containing lyophilized nucleic acid-silica adsorption buffer and silica-coated magnetic beads. After mixing, the sample was transferred from the pipette bulb to a small diameter tube, and captured biomarkers were concentrated using magnetic entrainment of beads through pre-arrayed wash solutions separated by small air gaps. Feasibility was tested using synthetic segments of the 140 bp tuberculosis IS6110 DNA sequence spiked into pooled human urine samples. DNA recovery was evaluated by qPCR. Despite the presence of spiked DNA, no DNA was detectable in unextracted urine samples, presumably due to the presence of PCR inhibitors. However, following extraction with the magnetic bead-based method, we found that ∼50% of spiked TB DNA was recovered from human urine containing roughly 5×10(3) to 5×10(8) copies of IS6110 DNA. In addition, the DNA was concentrated approximately ten-fold into water. The final concentration of DNA in the eluate was 5×10(6), 14×10(6), and 8×10(6) copies/µL for 1, 3, and 5 mL urine samples, respectively. Lyophilized and freshly prepared reagents within the transfer pipette produced similar results, suggesting that long-term storage without refrigeration is possible. DNA recovery increased with the length of the spiked DNA segments from 10±0.9% for a 75 bp DNA sequence to 42±4% for a 100 bp segment and 58±9% for a 140 bp segment. The estimated LOD was 77 copies of DNA/µL of urine. The strategy presented here provides a simple means to achieve high nucleic acid recovery from easily obtained urine samples, which does not contain inhibitors of PCR.

  10. A magnetic bead-based method for concentrating DNA from human urine for downstream detection.

    Directory of Open Access Journals (Sweden)

    Hali Bordelon

    Full Text Available Due to the presence of PCR inhibitors, PCR cannot be used directly on most clinical samples, including human urine, without pre-treatment. A magnetic bead-based strategy is one potential method to collect biomarkers from urine samples and separate the biomarkers from PCR inhibitors. In this report, a 1 mL urine sample was mixed within the bulb of a transfer pipette containing lyophilized nucleic acid-silica adsorption buffer and silica-coated magnetic beads. After mixing, the sample was transferred from the pipette bulb to a small diameter tube, and captured biomarkers were concentrated using magnetic entrainment of beads through pre-arrayed wash solutions separated by small air gaps. Feasibility was tested using synthetic segments of the 140 bp tuberculosis IS6110 DNA sequence spiked into pooled human urine samples. DNA recovery was evaluated by qPCR. Despite the presence of spiked DNA, no DNA was detectable in unextracted urine samples, presumably due to the presence of PCR inhibitors. However, following extraction with the magnetic bead-based method, we found that ∼50% of spiked TB DNA was recovered from human urine containing roughly 5×10(3 to 5×10(8 copies of IS6110 DNA. In addition, the DNA was concentrated approximately ten-fold into water. The final concentration of DNA in the eluate was 5×10(6, 14×10(6, and 8×10(6 copies/µL for 1, 3, and 5 mL urine samples, respectively. Lyophilized and freshly prepared reagents within the transfer pipette produced similar results, suggesting that long-term storage without refrigeration is possible. DNA recovery increased with the length of the spiked DNA segments from 10±0.9% for a 75 bp DNA sequence to 42±4% for a 100 bp segment and 58±9% for a 140 bp segment. The estimated LOD was 77 copies of DNA/µL of urine. The strategy presented here provides a simple means to achieve high nucleic acid recovery from easily obtained urine samples, which does not contain inhibitors of PCR.

  11. Inhibition of uropathogenic biofilm growth on silicone rubber in human urine by lactobacilli - a teleologic approach

    NARCIS (Netherlands)

    Velraeds, MMC; van de Belt-Gritter, B; Busscher, HJ; Reid, G; van der Mei, HC

    2000-01-01

    The ability of three Lactobacillus strains to inhibit the adhesion and growth of naturally occurring uropathogens on silicone rubber was investigated in human urine. The importance of biosurfactant production by Lactobacillus in discouraging uropathogen growth was determined in relation to the bindi

  12. Acute maternal rehydration increases the urine production rate in the near-term human fetus

    NARCIS (Netherlands)

    Haak, MC; Aarnoudse, JG; Oosterhof, H.

    OBJECTIVE: We sought to investigate the effect of a decrease of maternal plasma osmolality produced by hypotonic rehydration on the fetal urine production rate in normal near-term human fetuses. STUDY DESIGN: Twenty-one healthy pregnant women attending the clinic for antenatal care were studied

  13. Application of ion mobility spectrometry for the detection of human urine.

    Science.gov (United States)

    Rudnicka, Joanna; Mochalski, Paweł; Agapiou, Agapios; Statheropoulos, Milt; Amann, Anton; Buszewski, Bogusław

    2010-11-01

    The aim of the present study was to evaluate the suitability of ion mobility spectrometry (IMS) for the detection of human urine as an indication of human presence during urban search and rescue operations in collapsed buildings. To this end, IMS with a radioactive ionization source and a multicapillary column was used to detect volatile organic compounds (VOCs) emitted from human urine. A study involving a group of 30 healthy volunteers resulted in the selection of seven volatile species, namely acetone, propanal, 3-methyl-2-butanone, 2-methylpropanal, 4-heptanone, 2-heptanone and octanal, which were detected in all samples. Additionally, a preliminary study on the permeation of urine volatiles through the materials surrounding the voids of collapsed buildings was performed. In this study, quartz sand was used as a representative imitating material. Four compounds, namely 3-methyl-2-butanone, octanal, acetone and 2-heptanone, were found to permeate through the sand layers during all experiments. Moreover, their permeation times were the shortest. Although IMS can be considered as a potential technique suitable for the detection, localization and monitoring of VOCs evolved from human urine, further investigation is necessary prior to selecting field chemical methods for the early location of trapped victims.

  14. A Rapid and Sensitive Method for the Quantitation of Legionella Pneumophila Antigen from Human Urine.

    Science.gov (United States)

    1980-11-12

    reverse% passive hemagglutination test was developed to assay concentrations of solubl4 antigen of Legionnaires’ Disease ( Legionella pneumophila ) in... Legionella pneumophila Antigen from Humanl Urine JOSEPH A. MANGIAFICO, KENNETH W. HEDLUND, AND ALLEN R. KNOTT Running title: L. PNEUMOPHILA ANTIGEN IN...Approved for public release; distribution unlimited A Rapid and Sensitive Method for the Quantitation of Legionella pneumophila Antigen from Human

  15. Inhibition of uropathogenic biofilm growth on silicone rubber in human urine by lactobacilli - a teleologic approach

    NARCIS (Netherlands)

    Velraeds, MMC; van de Belt-Gritter, B; Busscher, HJ; Reid, G; van der Mei, HC

    2000-01-01

    The ability of three Lactobacillus strains to inhibit the adhesion and growth of naturally occurring uropathogens on silicone rubber was investigated in human urine. The importance of biosurfactant production by Lactobacillus in discouraging uropathogen growth was determined in relation to the bindi

  16. Characterization of a scyllo-inositol-containing sialyloligosaccharide from normal human urine.

    Science.gov (United States)

    Parkkinen, J

    1983-10-31

    Three inositol-containing sialyloligosaccharides were isolated from normal human urine. Structural studies including gas-liquid chromatography of mono- and disaccharide derivatives, methylation analysis, mass spectrometry and glycosidase treatments indicated the structure NeuAc (alpha 2-3)Gal(beta 1-0)scyllo-inositol for one of the oligosaccharides isolated. This provides the first evidence for the natural occurrence of a scyllo-inositol glycoside in biological material. The two other oligosaccharides isolated were identified as two isomers of NeuAc(alpha 2-3)Gal(beta 1-0)myo-inositol, which have not been identified in normal urine before.

  17. Application of duckweed for human urine treatment in Bioregenerative Life Support System

    Science.gov (United States)

    Manukovsky, Nickolay; Kovalev, Vladimir

    The object of the study was the common duckweed Lemna minor L. Thanks to the ability to assimilate mineral and organic substances, duckweed is used to purify water in sewage lagoons. In addition, duckweed biomass is known to be a potential high-protein feed resource for domestic animals and fish. The aim of the study was to estimate an application of duckweed in a two-stage treatment of human urine in Bioregenerative Life Support System (BLSS). At the first stage, the urine’s organic matter is oxidized by hydrogen peroxide. Diluted solution of oxidized urine is used for cultivation of duckweed. The appointment of duckweed is the assimilation of mineralized substances of urine. Part of the duckweed biomass yield directly or after composting could be embedded in the soil-like substrate as organic fertilizer to compensate the carry-over in consequence of plant growing. The rest duckweed biomass could be used as a feed for animals in BLSS. Then, the residual culture liquid is concentrated and used as a source of dietary salt. It takes 10-15 m2 of duckweed culture per crewmember to treat oxidized urine. The BLSS configuration including two-component subsystem of urine treatment is presented.

  18. Urban nutrient recovery from fresh human urine through cultivation of Chlorella sorokiniana.

    Science.gov (United States)

    Zhang, Shanshan; Lim, Chun Yong; Chen, Chia-Lung; Liu, He; Wang, Jing-Yuan

    2014-12-01

    High rate food consumption in urban cities causes vast amounts of nitrogen and phosphorus used in agriculture to end up in urban wastewaters. To substantially recover these nutrients, source-separated human urine should be targeted. The present study was to investigate the feasibility of recovering nitrogen and phosphorus in urine via microalgae cultivation. In concentrated urine, urea hydrolysis and precipitation occur rapidly, making microalgal growth difficult and nutrient recovery ineffective. However, when fresh urine was added as nutrient stock for 1-day growth requirement, biomass of Chlorella sorokiniana grew from 0.44 to 0.96 g L(-1) utilising 62.64 mg L(-1) of N and 10.64 mg L(-1) of P, achieving 80.4% and 96.6% recoveries, respectively in a 1-day non-sterile cultivation cycle. Overall, microalgae grown with urine added as nutrient supplement show no signs of inferiority as compared to those grown in recipe medium BG11 in terms of mass and chlorophyll a growth rates as well as resulting lipids (36.8%) and energy contents (21.0 kJ g(-1)).

  19. Detection of Volatile Metabolites Derived from Garlic (Allium sativum in Human Urine

    Directory of Open Access Journals (Sweden)

    Laura Scheffler

    2016-12-01

    Full Text Available The metabolism and excretion of flavor constituents of garlic, a common plant used in flavoring foods and attributed with several health benefits, in humans is not fully understood. Likewise, the physiologically active principles of garlic have not been fully clarified to date. It is possible that not only the parent compounds present in garlic but also its metabolites are responsible for the specific physiological properties of garlic, including its influence on the characteristic body odor signature of humans after garlic consumption. Accordingly, the aim of this study was to investigate potential garlic-derived metabolites in human urine. To this aim, 14 sets of urine samples were obtained from 12 volunteers, whereby each set comprised one sample that was collected prior to consumption of food-relevant concentrations of garlic, followed by five to eight subsequent samples after garlic consumption that covered a time interval of up to 26 h. The samples were analyzed chemo-analytically using gas chromatography-mass spectrometry/olfactometry (GC-MS/O, as well as sensorially by a trained human panel. The analyses revealed three different garlic-derived metabolites in urine, namely allyl methyl sulfide (AMS, allyl methyl sulfoxide (AMSO and allyl methyl sulfone (AMSO2, confirming our previous findings on human milk metabolite composition. The excretion rates of these metabolites into urine were strongly time-dependent with distinct inter-individual differences. These findings indicate that the volatile odorant fraction of garlic is heavily biotransformed in humans, opening up a window into substance circulation within the human body with potential wider ramifications in view of physiological effects of this aromatic plant that is appreciated by humans in their daily diet.

  20. Detection of Volatile Metabolites Derived from Garlic (Allium sativum) in Human Urine

    Science.gov (United States)

    Scheffler, Laura; Sauermann, Yvonne; Heinlein, Anja; Sharapa, Constanze; Buettner, Andrea

    2016-01-01

    The metabolism and excretion of flavor constituents of garlic, a common plant used in flavoring foods and attributed with several health benefits, in humans is not fully understood. Likewise, the physiologically active principles of garlic have not been fully clarified to date. It is possible that not only the parent compounds present in garlic but also its metabolites are responsible for the specific physiological properties of garlic, including its influence on the characteristic body odor signature of humans after garlic consumption. Accordingly, the aim of this study was to investigate potential garlic-derived metabolites in human urine. To this aim, 14 sets of urine samples were obtained from 12 volunteers, whereby each set comprised one sample that was collected prior to consumption of food-relevant concentrations of garlic, followed by five to eight subsequent samples after garlic consumption that covered a time interval of up to 26 h. The samples were analyzed chemo-analytically using gas chromatography-mass spectrometry/olfactometry (GC-MS/O), as well as sensorially by a trained human panel. The analyses revealed three different garlic-derived metabolites in urine, namely allyl methyl sulfide (AMS), allyl methyl sulfoxide (AMSO) and allyl methyl sulfone (AMSO2), confirming our previous findings on human milk metabolite composition. The excretion rates of these metabolites into urine were strongly time-dependent with distinct inter-individual differences. These findings indicate that the volatile odorant fraction of garlic is heavily biotransformed in humans, opening up a window into substance circulation within the human body with potential wider ramifications in view of physiological effects of this aromatic plant that is appreciated by humans in their daily diet. PMID:27916960

  1. Research Article. Perfluoroalkylated substances in human urine: results of a biomonitoring pilot study

    Directory of Open Access Journals (Sweden)

    Hartmann Christina

    2017-04-01

    Full Text Available Perfluoroalkylated substances (PFASs are a class of synthetic chemicals used in a wide range of processes and products due to their unique physicalchemical properties. Through intake of PFASs via food or several consumer products, humans can be exposed. Long-chain PFASs have been associated with adverse effects in laboratory animals, and there is also evidence for adverse health effects in humans. Although investigations of human exposure are mainly conducted in blood samples, some studies have shown that especially short-chain PFASs can be detected in human urine. In the present study, a sensitive analytical method was adapted for the measurement of 12 PFASs in human urine samples by HPLC-MS/MS. For verifying this method, concentrations in 11 male and female participants aged 25-46 years were analysed. In the study population, ranges of urinary PFASs concentrations were n.d.- 8.5 ng/l for perfluoropentanoic acid, human urine.

  2. Evaluation of a human on-site urine multidrug test for emergency use with dogs.

    Science.gov (United States)

    Teitler, Joan B

    2009-01-01

    A rapid, human on-site urine multidrug test was used to screen canine urine samples for the presence of five illegal drugs and drugs from three commonly abused drug classes. Each sample was sent to a toxicology laboratory for gas chromatography/mass spectrometry (GC/MS) validation. On-site test results and GC/MS assays confirmed that the human on-site test kit did identify barbiturates, opiates, benzodiazepines, and amphetamines/methamphetamines in urine from dogs that had received these common illicit drugs/drug classes either intravenously and/or orally. However, neither the on-site test kit nor the GC/MS individual assays for marijuana or methadone, a synthetic opiate, were effective in identifying marijuana and methadone in urine from dogs with suspected or known exposure. No index of suspicion was seen for exposure to phencyclidines or cocaine during the study period, and no exposures were indicated by the on-site test results. Overall, the test is a rapid, readily available, affordable, and useful complement to the veterinarian's clinical consideration and professional judgment.

  3. Technical note: effects of forage protein-binding polyphenols on chemistry of dairy excreta.

    Science.gov (United States)

    Powell, J M; Broderick, G A; Grabber, J H; Hymes-Fecht, U C

    2009-04-01

    Forage chemistry can affect intake, digestion, milk production, and manure excretion. Although information is available on the effects of forage protein-binding polyphenols on small ruminant production and manure excretion, little information is available for dairy cattle. The objective of this study was to compare fecal and urinary N excretion of diets formulated with alfalfa (Medicago sativa L.) silage versus condensed tannin-containing birdsfoot trefoil (Lotus corniculatus) or o-quinone-containing red clover (Trifolium pratense L.) silages. Significantly higher concentrations of N were excreted in urine by lactating Holstein dairy cows fed red clover and low-tannin birdsfoot trefoil (8.2 g/L) than by cows fed high-tannin birdsfoot trefoil or alfalfa (7.1 g/L). Fecal N concentrations were similar (33.6 g/kg) among all diets. Dairy cows fed red clover had lower rates of urinary N excretion (5.0 g/h) compared with other forages (6.6 g/h). Fecal N excretion rates were lowest for red clover (4.1 g/h), intermediate for alfalfa (5.8 g/h), and greatest for cows fed high- and low-tannin birdsfoot trefoil (6.4 g/h). The ratio of fecal N to urinary N was highest for high-tannin trefoil, lowest for alfalfa and red clover, and higher in excreta collected in morning than evening. Concentrations of neutral detergent fiber (NDF) in feces, of N in NDF (NDIN) and acid detergent fiber (ADIN), and relative amounts of NDIN and ADIN excreted in feces were significantly higher from cows fed high-tannin birdsfoot trefoil than the other silage types. Study results imply that collection of excreta for environmental studies needs to consider forage polyphenol and diurnal effects on chemistry of dairy excreta.

  4. Direct solid-phase microextraction combined with gas and liquid chromatography for the determination of lidocaine in human urine

    NARCIS (Netherlands)

    Koster, E.H M; Hofman, N.S K; de Jong, G.J.

    1998-01-01

    Solid-phase microextraction (SPME) has been combined with gas chromatography (GC) and liquid chromatography (LC) for the determination of lidocaine in human urine. A polydimethylsiloxane (PDMS) coated fibre was directly immersed into buffered urine. Extraction conditions such as time, pH, ionic stre

  5. Isolation and characterization of novel phosphate-containing sialyloligosaccharides from normal human urine.

    Science.gov (United States)

    Parkkinen, J; Finne, J

    1984-04-16

    Three phosphate-containing sialyloligosaccharides were isolated from normal human urine using charcoal adsorption, gel-filtration chromatography, ion-exchange chromatography and paper chromatography. Studies including gas-liquid chromatography of monosaccharide and disaccharide derivatives, methylation analysis, phosphate determination, ion-exchange chromatography and glycosidase and phosphatase treatments indicated the following three structures for the compounds isolated: NeuAc(alpha 2-6)Gal(beta 1-4)GlcNAc(alpha)-P; NeuAc(alpha 2-3)Gal(beta 1-4)GlcNAc(alpha)-P; NeuAc(alpha 2-3)Gal(beta 1-3)GalNAc(alpha)-P. These sialyloligosaccharide 1-phosphates represent a novel class of oligosaccharides. Their oligosaccharide chains are identical with the common sialyloligosaccharide end groups of glycoproteins and glycolipids. The excretion of these compounds in normal human urine may indicate the existence of a novel, as yet unrevealed pathway in the metabolism of complex carbohydrates.

  6. Novel validated spectrofluorimetric methods for the determination of taurine in energy drinks and human urine.

    Science.gov (United States)

    Sharaf El Din, M K; Wahba, M E K

    2015-03-01

    Two sensitive, selective, economic and validated spectrofluorimetric methods were developed for the determination of taurine in energy drinks and spiked human urine. Method Ι is based on fluorimetric determination of the amino acid through its reaction with Hantzsch reagent to form a highly fluorescent product measured at 490 nm after excitation at 419 nm. Method ΙΙ is based on the reaction of taurine with tetracyanoethylene yielding a fluorescent charge transfer complex, which was measured at λex /em of (360 nm/450 nm). The proposed methods were subjected to detailed validation procedures, and were statistically compared with the reference method, where the results obtained were in good agreement. Method Ι was further applied to determine taurine in energy drinks and spiked human urine giving promising results. Moreover, the stoichiometry of the reactions was studied, and reaction mechanisms were postulated.

  7. Rapid determination of vitamin B2 and B12 in human urine by isocratic liquid chromatography.

    Science.gov (United States)

    Mandal, Santi M; Mandal, Mahitosh; Ghosh, Ananta K; Dey, Satyahari

    2009-04-27

    A simple and rapid method for the identification and quantification of vitamin B(2) and B(12) in human urine has been developed using reverse phase high performance liquid chromatography (HPLC) and the peaks identity were confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). HPLC separation was performed in single wavelength detector (lambda(365)) mode and separated isocratically using mobile phase methanol: 1mM aqueous TFA (1:4) in C18 column. The calibration graphs plotted with different concentrations of vitamin B(2) and B(12) was linear with a correlation coefficients (r(2))=0.9975 and 0.9985, respectively. The recoveries of vitamin B(2) and B(12) were above 87% and 90%, respectively. The results of this present study suggest that the proposed method may be simple and convenient way of identifying and quantifying vitamin B(2) and B(12) from human urine.

  8. Comparison of Uric Acid Quantity with Different Food in Human Urine by Flow Injection Chemiluminescence Analysis

    Directory of Open Access Journals (Sweden)

    Jiajia Wang

    2013-01-01

    Full Text Available Based on the inhibitory effect of uric acid (UA on luminol-Co2+ chemiluminescence (CL system, a sensitive method for the determination of UA at nanomolar level by flow injection (FI CL was proposed. The proposed method was successfully applied to real-time monitoring of UA excretion in human 24 h urine with different food intake, showing that meats, vegetables, and porridge intake caused differential UA excretions of 879, 798, and 742 mg, respectively. It was also found that UA concentrations in urine under the three kinds of food intake simultaneously reached maximum at 2 h after meals with the values of 417, 318, and 288 μg mL−1, respectively. The UA concentration in human serum was also determined by this approach, and the possible mechanism of luminol-Co2+-UA CL reaction was discussed in detail.

  9. High-performance liquid chromatographic method for the determination of drotaverine in human plasma and urine.

    Science.gov (United States)

    Bolaji, O O; Onyeji, C O; Ogungbamila, F O; Ogunbona, F A; Ogunlana, E O

    1993-12-08

    A simple and sensitive HPLC method for the determination of drotaverine in human plasma and urine has been developed. Alkalinized plasma or urine was extracted with organic solvent and the basic components in the organic phase were back-extracted into 0.1 M HCl. An aliquot of the aqueous layer was injected onto the column and the eluent was monitored at 254 nm. Separation was performed on a C18-column with 0.02 M sodium dihydrogen phosphate-methanol (30:70, v/v) containing perchlorate ion at pH 3.2 as mobile phase. Drotaverine was well resolved from the plasma constituents and internal standard. An excellent linearity was observed between peak-height ratios and plasma concentrations and the intra- and inter-assay coefficients of variation were always drotaverine in humans as well as in animal models.

  10. [Determination of amphetamines in human urine using microwave extraction-gas chromatography].

    Science.gov (United States)

    Wang, Jifen; Sun, Hongfeng; Ye, Nengsheng; Gu, Xuexin; Li, Wenjun; Li, Ying

    2008-03-01

    A method has been developed for the determination of methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA) and 3,4-methylenedioxymethamphetamine (MDMA) in human urine using microwave extraction-gas chromatography(GC). To improve the extraction efficiency, experimental parameters of the extraction, including extraction solvent and its amount, pH value of the urine sample, extraction time and temperature were investigated. The optimal conditions were as follows: the pH value of urine sample at pH 12, cyclohexane as extraction solvent, extraction at 40 degrees C for 10 min. The average recoveries of MA, MDA and MDMA with this extraction method were 92.25%, 85.94% and 91.50%, the relative standard deviations were 5.5%, 5.5% and 6.1%(n = 5), and the limits of detection were 10, 20 and 20 ng/mL, respectively. Using this method, MA, MDA and MDMA need not be derivatized and can be separated from the matrix. The results indicate that the developed method is rapid, accurate and sensitive, and can be used for the simultaneous determination of MA, MDA and MDMA in urine samples.

  11. A salting out and resin procedure for extracting Schistosoma mansoni DNA from human urine samples

    Directory of Open Access Journals (Sweden)

    Rodrigues Nilton B

    2010-04-01

    Full Text Available Abstract Background In this paper a simple and cheap salting out and resin (InstaGene matrix® resin - BioRad DNA extraction method from urine for PCR assays is introduced. The DNA of the fluke Schistosoma mansoni was chosen as the target since schistosomiasis lacks a suitable diagnostic tool which is sensitive enough to detect low worm burden. It is well known that the PCR technique provides high sensitivity and specificity in detecting parasite DNA. Therefore it is of paramount importance to take advantage of its excellent performance by providing a simple to handle and reliable DNA extraction procedure, which permits the diagnosis of the disease in easily obtainable urine samples. Findings The description of the extraction procedure is given. This extraction procedure was tested for reproducibility and efficiency in artificially contaminated human urine samples. The reproducibility reached 100%, showing positive results in 5 assay repetitions of 5 tested samples each containing 20 ng DNA/5 ml. The efficiency of the extraction procedure was also evaluated in a serial dilution of the original 20 ng DNA/5 ml sample. Detectable DNA was extracted when it was at a concentration of 1.28 pg DNA/mL, revealing the high efficiency of this procedure. Conclusions This methodology represents a promising tool for schistosomiasis diagnosis utilizing a bio-molecular technique in urine samples which is now ready to be tested under field conditions and may be applicable to the diagnosis of other parasitic diseases.

  12. A New Molecularly Imprinted Polymer for Solid-phase Extraction of Cotinine from Human Urine

    Institute of Scientific and Technical Information of China (English)

    Jun YANG; Xiao Lan ZHU; Ji Bao CAI; Qing De SU; Yun GAO; Liang ZHANG

    2005-01-01

    A molecularly imprinted polymer (MIP), prepared around a cotinine template, has been synthesized. The feasibility of using the polymer for solid-phase extraction (SPE) of cotinine from biological samples has been investigated. The results show that cotinine can be quantitatively retained and eluted from the polymer. Experiments with human urine samples indicate that clean target analyte is obtained for HPLC with UV detection using the protocol.

  13. Biotransformation of aesculin by human gut bacteria and identification of its metabolites in rat urine

    Institute of Scientific and Technical Information of China (English)

    Wei-Jun Ding; Yun Deng; Hao Feng; Wei-Wei Liu; Rong Hu; Xiang Li; Zhe-Ming Gu; Xiao-Ping Dong

    2009-01-01

    AIM:To observe the biotransformation process of a Chinese compound, aesculin, by human gut bacteria, and to identify its metabolites in rat urine. METHODS:Representative human gut bacteria were collected from 20 healthy volunteers, and then utilized in vitro to biotransform aesculin under anaerobic conditions. At 0, 2, 4, 8, 12, 16, 24, 48 and 72 h postincubation, 10 mL of culture medium was collected. Metabolites of aesculin were extracted 3 × from rat urine with methanol and analyzed by HPLC. For in vivo metabolite analysis, aesculetin (100 mg/kg) was administered to rats via stomach gavage, rat urine was collected from 6 to 48 h post-administration, and metabolite analysis was performed by LC/ESI-MS and MS/MS in the positive and negative modes. RESULTS:Human gut bacteria could completely convert aesculin into aesculetin in vitro. The biotransformation process occurred from 8 to 24 h post-incubation, with its highest activity was seen from 8 to 12 h. The in vitro process was much slower than the in vivo process. In contrast to the in vitro model, six aesculetin metabolites were identified in rat urine, including 6-hydroxy-7-glucocoumarin (M1), 6-hydroxy-7-sulf-coumarin (M2), 6, 7-digluco- coumarin (M3), 6-glc-7-gluco-coumarin (M4), 6-O-methyl-7-gluco-coumarin (M5) and 6-O-methyl-7- sulf-coumarin (M6). Of which, M2 and M6 were novel metabolites. CONCLUSION:Aesculin can be transferred into aesculetin by human gut bacteria and is furth er modifiedby the host in vivo. The diverse metabolites of aesculin may explain its pleiotropic pharmaceutical effects.

  14. Application of thermoresponsive HPLC to forensic toxicology: determination of barbiturates in human urine

    OpenAIRE

    Kanno, Sanae; Watanabe, Kanako; HIRANO, SEISHIRO; Yamagishi, Itaru; Gonmori, Kunio; Minakata, Kayoko; Suzuki, Osamu

    2009-01-01

    A high-performance liquid chromatography (HPLC) method has been developed for the assays of five barbiturates in human urine using a new thermoresponsive polymer separation column, which is composed of N-isopropylacrylamide polymer. According to elevating the column temperature from 10 ℃ to 50 ℃, five barbiturates, such as metharbital, primidone, phenobarbital, mephobarbital and pentobarbital, became well separated by this method. Five barbiturates showed good linearity in the range of 0.2-10...

  15. Ion-pair HPLC method for the quantification of metformin in human urine

    Directory of Open Access Journals (Sweden)

    Eva Troja

    2016-01-01

    Full Text Available The aim of this study was to develop and validate a selective and sensitive ion-pairing HPLC–UV method for the determination of metformin in human urine using a conventional reversed-phase column. The bioanalytical method was carried out using a RP-C18 column (250×4.0 mm; 5 μm. A mobile phase consisting of acetonitrile and 10 mM sodium phosphate buffer (pH=6.0, 30:70 v/v and sodium dodecyl sulfate (0.3% was pumped at an isocratic flow rate of 1.00 mL/minute and quantification was achieved at 236 nm using using a UV/VIS DAD. The calibration curves were linear (r2 >0.999 in the concentration ranges of 62.5–2000 μg/mL for metformin in urine. LOD and LOQ were found to be 12 μg/mL and 35 μg/mL, respectively. The method was found to be rapid, precise and accurate for quantification of metformin in human urine. This method was successfully applied to a pharmacokinetic study of humans through oral administration.

  16. Magnetic graphene oxide as adsorbent for the determination of polycyclic aromatic hydrocarbon metabolites in human urine.

    Science.gov (United States)

    Zhu, Linli; Xu, Hui

    2014-09-01

    Detection of monohydroxy polycyclic aromatic hydrocarbons metabolites in urine is an advisable and valid method to assess human environmental exposure to polycyclic aromatic hydrocarbons. In this work, novel Fe3O4/graphene oxide composites were prepared and their application in the magnetic solid-phase extraction of monohydroxy polycyclic aromatic hydrocarbons in urine was investigated by coupling with liquid chromatography and mass spectrometry. In the hybrid material, superparamagnetic Fe3O4 nanoparticles provide fast separation to simplify the analytical process and graphene oxide provides a large functional surface for the adsorption. The prepared magnetic nanocomposites were characterized by X-ray diffraction, scanning electron microscopy, transmission electron microscopy, and vibrating sample magnetometry. The experimental conditions were optimized systematically. Under the optimal conditions, the recoveries of these compounds were in the range of 98.3-125.2%, the relative standard deviations ranged between 6.8 and 15.5%, and the limits of detection were in the range of 0.01-0.15 ng/mL. The simple, quick, and affordable method was successfully used in the analysis of human urinary monohydroxy polycyclic aromatic hydrocarbons in two different cities. The results indicated that the monohydroxy polycyclic aromatic hydrocarbons level in human urine can provide useful information for environmental exposure to polycyclic aromatic hydrocarbons.

  17. The culture of Chlorella vulgaris with human urine in multibiological life support system experiments

    Science.gov (United States)

    Li, Ming; Liu, Hong; Tong, Ling; Fu, Yuming; He, Wenting; Hu, Enzhu; Hu, Dawei

    The Integrative Experimental System (IES) was established as a tool to evaluate the rela-tionship of the subsystems in Bioregenerative Life Support System, and Multibiological Life Support System Experiments (MLSSE) have been conducted in the IES. The IES consists of a higher plant chamber, an animal chamber and a plate photo bioreactor (PPB) which cultivated lettuce (Lactuca sativa L.), silkworm (Bombyx Mori L.) and microalgae (Chlorella vulgaris), respectively. In MLSSE, four volunteers took turns breathing the system air through a tube connected with the animal chamber periodically. According to the CO2 concentration in the IES, the automotive control system of the PPB changed the light intensity regulating the photosynthesis of Chlorella vulgaris to make CO2 /O2 in the system maintain at stable levels. Chlorella vulgaris grew with human urine by carrying certain amount of alga liquid out of the bioreactor every day with synthetic urine replenished into the system, and O2 was regenerated, at the same time human urine was purified. Results showed that this IES worked stably and Chlorella vulgaris grew well; The culture of Chlorella vulgaris could be used to keep the balance of CO2 and O2 , and the change of light intensity could control the gas composition in the IES; Microalgae culture could be used in emergency in the system, the culture of Chlorella vulgaris could recover to original state in 5 days; 15.6 ml of condensation water was obtained every day by the culture of Chlorella vulgaris; The removal efficiencies of N, P in human urine could reach to 98.2% and 99.5%.

  18. Semisolid liver infusion tryptose supplemented with human urine allows growth and isolation of Trypanosoma cruzi and Trypanosoma rangeli clonal lineages

    OpenAIRE

    2016-01-01

    Abstract: INTRODUCTION This work shows that 3% (v/v) human urine (HU) in semisolid Liver Infusion Tryptose (SSL) medium favors the growth of Trypanosoma cruzi and T. rangeli. METHODS Parasites were plated as individual or mixed strains on SSL medium and on SSL medium with 3% human urine (SSL-HU). Isolate DNA was analyzed using polymerase chain reaction (PCR) and pulsed-field gel electrophoresis (PFGE). RESULTS SSL-HU medium improved clone isolation. PCR revealed that T. cruzi strains pr...

  19. Evidence for Public Health Risks of Wastewater and Excreta Management Practices in Southeast Asia: A Scoping Review

    Directory of Open Access Journals (Sweden)

    Steven Lam

    2015-10-01

    Full Text Available The use of wastewater and excreta in agriculture is a common practice in Southeast Asia; however, concerns remain about the potential public health risks of this practice. We undertook a scoping review to examine the extent, range, and nature of literature, as well as synthesize the evidence for associations between wastewater and excreta management practices and public health risks in Southeast Asia. Three electronic databases (PubMed, CAB Direct, and Web of Science were searched and a total of 27 relevant studies were included and evaluated. The available evidence suggested that possible occupational health risks of wastewater and excreta management practices include diarrhea, skin infection, parasitic infection, bacterial infection, and epilepsy. Community members can be at risk for adverse health outcomes through consuming contaminated fish, vegetables, or fruits. Results suggested that practices including handling, treatment, and use of waste may be harmful to human health, particularly farmer’s health. Many studies in this review, however, had limitations including lack of gender analyses, exposure assessment, and longitudinal study designs. These findings suggest that more studies on identifying, quantitatively assessing, and mitigating health risks are needed if sustainable benefits are to be obtained from wastewater and excreta reuse in agriculture in Southeast Asia.

  20. Evidence for Public Health Risks of Wastewater and Excreta Management Practices in Southeast Asia: A Scoping Review

    Science.gov (United States)

    Lam, Steven; Nguyen-Viet, Hung; Tuyet-Hanh, Tran Thi; Nguyen-Mai, Huong; Harper, Sherilee

    2015-01-01

    The use of wastewater and excreta in agriculture is a common practice in Southeast Asia; however, concerns remain about the potential public health risks of this practice. We undertook a scoping review to examine the extent, range, and nature of literature, as well as synthesize the evidence for associations between wastewater and excreta management practices and public health risks in Southeast Asia. Three electronic databases (PubMed, CAB Direct, and Web of Science) were searched and a total of 27 relevant studies were included and evaluated. The available evidence suggested that possible occupational health risks of wastewater and excreta management practices include diarrhea, skin infection, parasitic infection, bacterial infection, and epilepsy. Community members can be at risk for adverse health outcomes through consuming contaminated fish, vegetables, or fruits. Results suggested that practices including handling, treatment, and use of waste may be harmful to human health, particularly farmer’s health. Many studies in this review, however, had limitations including lack of gender analyses, exposure assessment, and longitudinal study designs. These findings suggest that more studies on identifying, quantitatively assessing, and mitigating health risks are needed if sustainable benefits are to be obtained from wastewater and excreta reuse in agriculture in Southeast Asia. PMID:26501297

  1. Evidence for Public Health Risks of Wastewater and Excreta Management Practices in Southeast Asia: A Scoping Review.

    Science.gov (United States)

    Lam, Steven; Nguyen-Viet, Hung; Tuyet-Hanh, Tran Thi; Nguyen-Mai, Huong; Harper, Sherilee

    2015-10-15

    The use of wastewater and excreta in agriculture is a common practice in Southeast Asia; however, concerns remain about the potential public health risks of this practice. We undertook a scoping review to examine the extent, range, and nature of literature, as well as synthesize the evidence for associations between wastewater and excreta management practices and public health risks in Southeast Asia. Three electronic databases (PubMed, CAB Direct, and Web of Science) were searched and a total of 27 relevant studies were included and evaluated. The available evidence suggested that possible occupational health risks of wastewater and excreta management practices include diarrhea, skin infection, parasitic infection, bacterial infection, and epilepsy. Community members can be at risk for adverse health outcomes through consuming contaminated fish, vegetables, or fruits. Results suggested that practices including handling, treatment, and use of waste may be harmful to human health, particularly farmer's health. Many studies in this review, however, had limitations including lack of gender analyses, exposure assessment, and longitudinal study designs. These findings suggest that more studies on identifying, quantitatively assessing, and mitigating health risks are needed if sustainable benefits are to be obtained from wastewater and excreta reuse in agriculture in Southeast Asia.

  2. Determination of tenuazonic acid in human urine by means of a stable isotope dilution assay.

    Science.gov (United States)

    Asam, Stefan; Habler, Katharina; Rychlik, Michael

    2013-05-01

    The content of tenuazonic acid in human urine was determined by a stable isotope dilution assay (SIDA) that was recently developed for the analysis of food commodities and extensively re-validated for urine matrix in this study. Linearity of the response curve was proven between molar ratios n(labeled standard)/n(analyte) of 0.02-100. The limits of detection and determination were 0.2 and 0.6 μg/L, respectively. The mean recovery of the stable isotope dilution assay was 102 ± 3 % in the range between 1.0 and 100 μg/L. Interassay precision was 6.7 % (relative standard deviation of three triplicate analyses of a human urine sample during 3 weeks). The method was applied to two studies dealing with urinary excretion of tenuazonic acid: In the first study, tenuazonic acid was quantified in the 24-h urine of six volunteers from Germany (three female, three male) in a concentration range of 1.3-17.3 μg/L or 2.3-10.3 ng/mg(-1) creatinine, respectively. In the second study, two volunteers (one female, one male) ingested 30 μg tenuazonic acid by consumption of naturally contaminated whole meal sorghum infant cereals and tomato juice, respectively. The urinary excretion of the ingested tenuazonic acid was 54-81 % after 6 h, depending on matrix and volunteer. After 24 h, 87-93 % of the ingested amount of tenuazonic acid was excreted, but the fate of the remaining about 10 % is open. Thus, it is not possible to exclude potential health hazards for the consumer, completely.

  3. Determination of salbutamol and salbutamol glucuronide in human urine by means of liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Mareck, Ute; Guddat, Sven; Schwenke, Anne; Beuck, Simon; Geyer, Hans; Flenker, Ulrich; Elers, Jimmi; Backer, Vibeke; Thevis, Mario; Schänzer, Wilhelm

    2011-01-01

    The determination of salbutamol and its glucuronide in human urine following the inhalative and oral administration of therapeutic doses of salbutamol preparations was performed by means of direct urine injection utilizing liquid chromatography-tandem mass spectrometry (LC-MS/MS) and employing d(3)-salbutamol and d(3)-salbutamol glucuronide as internal standards. Unconjugated salbutamol was detected in all administration study urine samples. Salbutamol concentrations following inhalation were commonly (99%) below 1000 ng/ml whereas values after oral administration frequently (48%) exceeded this threshold. While salbutamol glucuronide was not detected in urine samples collected after inhalation of the drug, 26 out of 82 specimens obtained after oral application contained salbutamol glucuronide with a peak value of 63 ng/ml. The percentage of salbutamol glucuronide compared to unconjugated salbutamol was less than 3%. Authentic doping control urine samples indicating screening results for salbutamol less than 1000 ng/ml, showed salbutamol glucuronide concentrations between 2 and 6 ng/ml, whereas adverse analytical findings resulting from salbutamol levels higher than 1000 ng/ml, had salbutamol glucuronide values between 8 and 15 ng/ml. The approach enabled the rapid determination of salbutamol and its glucuronic acid conjugate in human urine and represents an alternative to existing procedures since time-consuming hydrolysis or derivatization steps were omitted. Moreover, the excretion of salbutamol glucuronide in human urine following the administration of salbutamol was proven.

  4. Anticlastogenic Effect of Redistilled Cow's Urine Distillate in Human Peripheral Lymphocytes Challenged With Manganese Dioxide and Hexavalent Chromium

    Institute of Scientific and Technical Information of China (English)

    DIPANWITA DUTTA; S.SARAVANA DEVI; K. KRISHNAMURTHI; T. CHAKRABARTI

    2006-01-01

    Objective To study the anticlastogenic effect of redistilled cow's urine distillate (RCUD) in human peripheral lymphocytes (HLC) challenged with manganese dioxide and hexavalent chromium. Methods The anticlastogenic activity of redistilled cow's urine distillate was studied in human polymorphonuclear leukocytes (HPNLs) and human peripheral lymphocytes in vitro challenged with manganese dioxide and hexavalent chromium as established genotoxicants and clastogens which could cause induction of DNA strand break, chromosomal aberration and micronucleus. Three different levels of RCUD: 1 μL/mL, 50 μL/mL and 100μL/mL, were used in the study. Results Manganese dioxide and hexavalent chromium caused statistically significant DNA strand break, chromosomal aberration and micronucleus formation, which could be protected by redistilled cow's urine distillate. Conclusion The redistilled cow's urine distillate posseses strong antigenotoxic and anticlastogenic properties against HPNLs and HLC treated with Cr+6 and MnO2. This property is mainly due to the antioxidants present in RCUD.

  5. Selective arsenic speciation analysis of human urine reference materials using gradient elution ion-exchange HPLC-ICP-MS

    DEFF Research Database (Denmark)

    Sloth, Jens Jørgen; Larsen, Erik Huusfeldt; Julshamn, K.

    2004-01-01

    Arsenic speciation analysis was performed in two human urine certified reference materials (NIES No. 18 and NIST SRM2670a) and three human urine control materials (Seronorm, Medisafe and Lyphocheck). The samples were diluted 1 + 3 prior to analysis by gradient elution anion or cation exchange high-performance...... liquid chromatography ( HPLC) coupled with inductively coupled plasma mass spectrometry (ICP-MS). Nine arsenic species, including arsenic acid, arsenous acid, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine, trimethylarsine oxide, dimethylarsinoylacetic acid, trimethylarsoniopropionate...... and dimethylarsinoylethanol, were determined in the urines. Additionally, several unknown arsenicals were detected. This is the first time that dimethylarsinoylacetic acid and trimethylarsoniopropionate have been reported in human urine. The sums of the species concentrations determined by the chromatographic approaches were...

  6. Human urine and wood ash as plant nutrients for red beet (Beta vulgaris) cultivation: impacts on yield quality.

    Science.gov (United States)

    Pradhan, Surendra K; Holopainen, Jarmo K; Weisell, Janne; Heinonen-Tanski, Helvi

    2010-02-10

    The objective of this study was to evaluate the effect of human urine and wood ash fertilization on the yield and quality of red beet by measuring the microbial, nutrient, and antioxidant (betanin) content of the roots. Red beets were fertilized with 133 kg of N/ha as mineral fertilizer, urine and ash, and only urine with no fertilizer as a control. The mineral-fertilized plants and urine- and ash-fertilized plants also received 89 kg of P/ha. Urine and ash and only urine fertilizer produced 1720 and 656 kg/ha more root biomass, respectively, versus what was obtained from the mineral fertilizer. Few fecal coliforms and coliphage were detected in mineral-fertilized and urine- and ash-fertilized red beet roots. The protein and betanin contents in red beet roots were similar in all treatments. In conclusion, this study revealed that urine with or without ash can increase the yield of red beet and furthermore the microbial quality and chemical quality were similar to the situation in mineral-fertilized products.

  7. Analysis of Fluconazole in Human Urine Sample by High Performance Liquid Chromatography Method

    Science.gov (United States)

    Hermawan, D.; Ali, N. A. Md; Ibrahim, W. A. Wan; Sanagi, M. M.

    2013-04-01

    A method for determination of fluconazole, antifungal drug in human urine by using reversed-phased high performance liquid chromatography (RP-HPLC) with ultraviolet (UV) detector was developed. Optimization HPLC conditions were carried out by changing the flow rate and composition of mobile phase. The optimum separation conditions at a flow rate 0.85 mL/min with a composition of mobile phase containing methanol:water (70:30, v/v) with UV detection at a wavelength 254 nm was able to analyze fluconazole within 3 min. The excellent linearity was obtained in the range of concentration 1 to 10 μg/mL with r2 = 0.998. The limit of detection (LOD) and limit of quantitation (LOQ) were 0.39 μg/mL and 1.28 μg/mL, respectively. Solid phase extraction (SPE) method using octadecylsilane (C18) as a sorbent was used to clean-up and pre-concentrated of the urine sample prior to HPLC analysis. The average recoveries of fluconazole in spiked urine sample was 72.4% with RSD of 3.21% (n=3).

  8. Extraction and Determination of Cyproheptadine in Human Urine by DLLME-HPLC Method.

    Science.gov (United States)

    Maham, Mehdi; Kiarostami, Vahid; Waqif-Husain, Syed; Abroomand-Azar, Parviz; Tehrani, Mohammad Saber; Khoeini Sharifabadi, Malihe; Afrouzi, Hossein; Shapouri, Mahmoudreza; Karami-Osboo, Rouhollah

    2013-01-01

    Novel dispersive liquid-liquid microextraction (DLLME), coupled with high performance liquid chromatography with photodiode array detection (HPLC-DAD) has been applied for the extraction and determination of cyproheptadine (CPH), an antihistamine, in human urine samples. In this method, 0.6 mL of acetonitrile (disperser solvent) containing 30 μL of carbon tetrachloride (extraction solvent) was rapidly injected by a syringe into 5 mL urine sample. After centrifugation, the sedimented phase containing enriched analyte was dissolved in acetonitrile and an aliquot of this solution injected into the HPLC system for analysis. Development of DLLME procedure includes optimization of some important parameters such as kind and volume of extraction and disperser solvent, pH and salt addition. The proposed method has good linearity in the range of 0.02-4.5 μg mL(-1) and low detection limit (13.1 ng mL(-1)). The repeatability of the method, expressed as relative standard deviation was 4.9% (n = 3). This method has also been applied to the analysis of real urine samples with satisfactory relative recoveries in the range of 91.6-101.0%.

  9. Determination of triamterene in human plasma and urine after its cloud point extraction

    Directory of Open Access Journals (Sweden)

    Ahad Bavili Tabrizi

    2014-01-01

    Full Text Available A new analytical approach was developed involving cloud point extraction (CPE and spectrofluorimetric determination of triamterene (TM in biological fluids. A urine or plasma sample was prepared and adjusted to pH 7, then TM was quickly extracted using CPE, using 0.05% (w/v of Triton X-114 as the extractant. The main factors that affected the extraction efficiency (the pH of the sample, the Triton X-114 concentration, the addition of salt, the extraction time and temperature, and the centrifugation time and speed were studied and optimized. The method gave calibration curves for TM with good linearities and correlation coefficients (r higher than 0.99. The method showed good precision and accuracy, with intra- and inter-assay precisions of less than 8.50% at all concentrations. Standard addition recovery tests were carried out, and the recoveries ranged from 94.7% to 114%. The limits of detection and quantification were 3.90 and 11.7 µg L-1, respectively, for urine and 5.80 and 18.0 µg L-1, respectively, for plasma. The newly developed, environmentally friendly method was successfully used to extract and determine TM in human urine samples.

  10. Identification of polyphenols and their metabolites in human urine after cranberry-syrup consumption.

    Science.gov (United States)

    Iswaldi, Ihsan; Arráez-Román, David; Gómez-Caravaca, Ana María; Contreras, María Del Mar; Uberos, José; Segura-Carretero, Antonio; Fernández-Gutiérrez, Alberto

    2013-05-01

    As the beneficial effects of American cranberry (Vaccinium macrocarpon) can be partly attributed to its phenolic composition, the evaluation of the physiological behaviour of this fraction is crucial. A rapid and sensitive method by ultra-performance liquid chromatography coupled to quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS) has been used to identify phenolic metabolites in human urine after a single dose of cranberry syrup. Prior to the analysis, metabolites were extracted using an optimised solid-phase extraction procedure. All possible metabolites were investigated based on retention time, accurate mass data and isotope and fragmentation patterns. Free coumaroyl hexose (isomer 1 and 2), dihydroxybenzoic acid, caffeoyl glucose, dihydroferulic acid 4-O-β-d-glucuronide, methoxyquercetin 3-O-galactoside, scopoletin, myricetin and quercetin, together with other 23 phase-I and phase-II metabolites, including various isomers, could be tentatively identified in the urine. Afterwards, the metabolites were simultaneously screened in the urine of different subjects at 0, 2, 4, and 6h after the ingestion of cranberry syrup by Target Analysis(TM) software.

  11. Urine proteomes of healthy aging humans reveal extracellular matrix (ECM) alterations and immune system dysfunction.

    Science.gov (United States)

    Bakun, M; Senatorski, G; Rubel, T; Lukasik, A; Zielenkiewicz, P; Dadlez, M; Paczek, L

    2014-02-01

    Aging is a complex physiological process that poses considerable conundrums to rapidly aging societies. For example, the risk of dying from cardiovascular diseases and/or cancer steadily declines for people after their 60s, and other causes of death predominate for seniors older than 80 years of age. Thus, physiological aging presents numerous unanswered questions, particularly with regard to changing metabolic patterns. Urine proteomics analysis is becoming a non-invasive and reproducible diagnostic method. We investigated the urine proteomes in healthy elderly people to determine which metabolic processes were weakened or strengthened in aging humans. Urine samples from 37 healthy volunteers aged 19-90 years (19 men, 18 women) were analyzed for protein expression by liquid chromatography-tandem mass spectrometry. This generated a list of 19 proteins that were differentially expressed in different age groups (young, intermediate, and old age). In particular, the oldest group showed protein changes reflective of altered extracellular matrix turnover and declining immune function, in which changes corresponded to reported changes in cardiovascular tissue remodeling and immune disorders in the elderly. Thus, urinary proteome changes in the elderly appear to reflect the physiological processes of aging and are particularly clearly represented in the circulatory and immune systems. Detailed identification of "protein trails" creates a more global picture of metabolic changes that occur in the elderly.

  12. Optimization of a Pre-MEKC Separation SPE Procedure for Steroid Molecules in Human Urine Samples

    Directory of Open Access Journals (Sweden)

    Ilona Olędzka

    2013-11-01

    Full Text Available Many steroid hormones can be considered as potential biomarkers and their determination in body fluids can create opportunities for the rapid diagnosis of many diseases and disorders of the human body. Most existing methods for the determination of steroids are usually time- and labor-consuming and quite costly. Therefore, the aim of analytical laboratories is to develop a new, relatively low-cost and rapid implementation methodology for their determination in biological samples. Due to the fact that there is little literature data on concentrations of steroid hormones in urine samples, we have made attempts at the electrophoretic determination of these compounds. For this purpose, an extraction procedure for the optimized separation and simultaneous determination of seven steroid hormones in urine samples has been investigated. The isolation of analytes from biological samples was performed by liquid-liquid extraction (LLE with dichloromethane and compared to solid phase extraction (SPE with C18 and hydrophilic-lipophilic balance (HLB columns. To separate all the analytes a micellar electrokinetic capillary chromatography (MECK technique was employed. For full separation of all the analytes a running buffer (pH 9.2, composed of 10 mM sodium tetraborate decahydrate (borax, 50 mM sodium dodecyl sulfate (SDS, and 10% methanol was selected. The methodology developed in this work for the determination of steroid hormones meets all the requirements of analytical methods. The applicability of the method has been confirmed for the analysis of urine samples collected from volunteers—both men and women (students, amateur bodybuilders, using and not applying steroid doping. The data obtained during this work can be successfully used for further research on the determination of steroid hormones in urine samples.

  13. Determination of lipoic acid in human urine by capillary zone electrophoresis.

    Science.gov (United States)

    Kubalczyk, Paweł; Głowacki, Rafał

    2017-07-01

    Fast, simple, and accurate CE method enabling determination of lipoic acid (LA) in human urine has been developed and validated. LA is a disulfide-containing natural compound absorbed from the organism's diet. Due to powerful antioxidant activity, LA has been used for prevention and treatment of various diseases and disorders, e.g. cardiovascular diseases, neurodegenerative disorders, and cancer. The proposed analytical procedure consists of liquid-liquid sample extraction, reduction of LA with tris(2-carboxyethyl)phosphine, derivatization with 1-benzyl-2-chloropyridinium bromide (BCPB) followed by field amplified sample injection stacking, capillary zone electrophoresis separation, and ultraviolet-absorbance detection of LA-BCPB derivative at 322 nm. Effective baseline electrophoretic separation was achieved within 6 min under the separation voltage of 20 kV (∼80 μA) using a standard fused-silica capillary (effective length 51.5 cm, 75 μm id) and BGE consisted of 0.05 mol/L borate buffer adjusted to pH 9. The experimentally determined limit of detection for LA in urine was 1.2 μmol/L. The calibration curve obtained for LA in urine showed linearity in the range 2.5-80 μmol/L, with R(2) 0.9998. The relative standard deviation of the points of the calibration curve was lower than 10%. The analytical procedure was successfully applied to analysis of real urine samples from seven healthy volunteers who received single 100 mg dose of LA. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Microbial health hazard assessment associated with the utilization of source-separated human urine%源分离尿液收集、利用的病原性健康风险计算评估

    Institute of Scientific and Technical Information of China (English)

    仇付国; 高红伟; 卢文灏

    2013-01-01

    为了明确源分离生态卫生系统收集、利用尿液过程中所导致的人体健康风险,对尿液中病原体的含量进行了调查分析,采用蒙特卡洛方法对尿液收集、利用环节所导致的健康风险进行了模拟计算.结果表明,尿液中的病原体主要来自粪便的交叉污染,细菌和隐孢子虫等不会造成大的风险,风险主要来源于病毒,即使摄人剂量很低(1mL)的、没处理过的尿液,其中的病毒也会对人体健康产生很高的风险(0.02~0.87 a-,均值为0.08 a-1).尿液浇灌施肥时造成的感染风险和尿液的储存时间有关,经过30 d的储存后,细菌和原生动物的尿液对人体的感染风险都很低,然而对于在5℃下储存6个月的尿液或20℃下储存1个月的尿液,轮状病毒的感染风险概率为10-1 a-1的数量级.如果尿液喷洒前在20℃下储存6个月,平均风险会降低到10-5 a-1数量级.对食用尿液浇灌的农产品进行的风险计算表明,浇灌两周后采摘食用蔬菜造成的健康风险为10-9 a-数量级,可以忽略不计.%The paper is to introduce a microbial health hazard assessment associated with the utilization of source-separated human urine.The reason why we would like to pick up the research topic is to make benefits of the urine utilization as a natural fertilizer in agriculture in favor of reducing the nitrogen and phosphorus load in the domestic sewage and transmitting human excreta with less fresh water in comparison with the conventional gravitational flow drainage system.However,it is necessary to identify the infectious hazards of disease when handling and using the urine so as to protect people' s health and promote the development of ecological sanitation environment.In this study,we have collected data of human faecal sterols in the urine and worked out the faeces content in it based on the average sterol level in them.Then we have done the evaluation of the amount of pathogen in the isolated urine by

  15. Preconcentration of indapamide from human urine using molecularly imprinted solid-phase extraction.

    Science.gov (United States)

    Yılmaz, Hüma; Basan, Hasan

    2015-09-01

    A simple, sensitive, and selective molecularly imprinted solid-phase extraction and spectrophotometric method has been developed for the clean-up and preconcentration of indapamide from human urine. Molecularly imprinted polymers were prepared by a non-covalent imprinting approach using indapamide as a template molecule, 2-(trifluoromethyl) acrylic acid as a functional monomer, ethylene glycol dimethacrylate as a crosslinker, N,N-azobisisobutyronitrile as a thermal initiator and acetonitrile as a porogenic solvent. A non-imprinted polymer was also prepared in the same way, but in the absence of template. Molecularly imprinted polymer and non-imprinted polymer sorbents were dry-packed into solid-phase extraction cartridges. Eluates from cartridges were analyzed using a spectrophotometer for the determination of indapamide by referring to the calibration curve in the range 0.14-1.50 μg/mL. Preconcentration factor, limit of detection, and limit of quantification were 16.30, 0.025 μg/mL, and 0.075 μg/mL, respectively. A relatively high imprinting factor (9.3) was also achieved and recovery values for the indapamide spiked into human urine were in the range of 80.1-81.2%. In addition, relatively low within-day (0.17-0.42%) and between-day (1.1-1.4%) precision values were obtained as well. The proposed molecularly imprinted solid-phase extraction and spectrophotometric method was successfully applied to selective extraction, preconcentration, and determination of indapamide from human urine samples.

  16. Exposure of family members to antineoplastic drugs via excreta of treated cancer patients.

    Science.gov (United States)

    Yuki, Michiko; Sekine, Satoko; Takase, Kanae; Ishida, Takashi; Sessink, Paul J M

    2013-09-01

    (a) To measure the urinary excretion of antineoplastic drugs of three patients during 48 h after the administration of cyclophosphamide (two patients) and 5-fluorouracil (one patient). (b) To evaluate environmental contamination with antineoplastic drugs via excreta of patients in the home setting. (c) To evaluate exposure of family members to antineoplastic drugs by measuring the drugs in their urine during the 48 h after completion of the chemotherapy by the patients. Two patients were administered cyclophosphamide by i.v. bolus injection. One patient was administered 5-fluorouracil by i.v. bolus injection and thereafter immediately administered the same drug by continuous infusion for 46 h. Urine samples from the patients administered cyclophosphamide and their family members, and wipe samples from their home environment, were analysed for the unchanged form of cyclophosphamide. For 5-fluorouracil, the urine samples from the patient and the family member were analysed for the 5-fluorouracil metabolite α-fluoro-β-alanine. Wipe samples were analysed for 5-fluorouracil. Drugs were detected and quantified with gas chromatography in tandem with mass spectroscopy-mass spectroscopy or by high-performance liquid chromatography with ultraviolet-light detection. A total of 35 and 16 urine samples were collected from the three patients and their family members, respectively. The drugs were detected in all samples. Cyclophosphamide was detected at levels of 0.03-7.34 ng/cm(2) in 8 of the 12 wipe samples obtained from the homes of the patients administered cyclophosphamide. For the patient administered 5-fluorouracil, drug levels in his home environment were below the limit of detection. We demonstrated contamination of the home setting and exposure of family members to cyclophosphamide via the excreta of outpatient receiving chemotherapy. Exposure of the family member of the patient administered 5-fluorouracil was also demonstrated. These findings indicate the

  17. Validation of a standardised method for determining beryllium in human urine at nanogram level.

    Science.gov (United States)

    Devoy, Jérôme; Melczer, Mathieu; Antoine, Guillaume; Remy, Aurélie; Heilier, Jean-François

    2013-10-01

    The potential toxicity of beryllium at low levels of exposure means that a biological and/or air monitoring strategy may be required to monitor the exposure of subjects. The main objective of the work presented in this manuscript was to develop and validate a sensitive and reproducible method for determining levels of beryllium in human urine and to establish reference values in workers and in non-occupationally exposed people. A chelate of beryllium acetylacetonate formed from beryllium(II) in human urine was pre-concentrated on a SPE C18 cartridge and eluted with methanol. After drying the eluate, the residue was solubilised in nitric acid and analysed by atomic absorption spectrometry and/or inductively coupled plasma mass spectrometry. The proposed method is 4 to 100 times more sensitive than other methods currently in routine use. The new method was validated with the concordance correlation coefficient test for beryllium concentrations ranging from 10 to 100 ng/L. Creatinine concentration, urine pH, interfering compounds and freeze-thaw cycles were found to have only slight effects on the performance of the method (less than 6%). The effectiveness of the two analytical techniques was compared statistically with each other and to direct analysis techniques. Even with a detection limit of 0.6 ng/L (obtained with inductively coupled plasma mass spectrometry), the method is not sensitive enough to detect levels in non-occupationally exposed persons. The method performance does however appear to be suitable for monitoring worker exposure in some industrial settings and it could therefore be of use in biological monitoring strategies.

  18. Flow method based on cloud point extraction for fluorometric determination of epinephrine in human urine.

    Science.gov (United States)

    Davletbaeva, Polina; Falkova, Marina; Safonova, Evgenia; Moskvin, Leonid; Bulatov, Andrey

    2016-03-10

    A novel stepwise injection fluorometric method for the determination of epinephrine in human urine has been developed. In the current study, the stepwise injection analysis (SWIA) was successfully combined with on-line in-syringe cloud point extraction (CPE) and fluorometric detection. The procedure was based on the epinephrine derivatization in the presence of o-phenylenediamine followed by the preconcentration stage based on the CPE with the nonionic surfactant Triton X-114. After the phase separation into a syringe of the flow system, the micellar phase containing the epinephrine derivative was transported to a fluorometric detector. The excitation and emission wavelengths were set at 447 nm and 550 nm, respectively. The conditions of epinephrine derivatization and CPE have been studied. The calibration plot constructed using the developed procedure was linear in the range of 1·10(-11)-5·10(-7) mol L(-1). The limit of detection, calculated as 3 σ of a blank test (n = 10), was found to be 3·10(-12) mol L(-1). The proposed method was successfully applied for the determination of epinephrine in human urine samples.

  19. Siderophore biosynthesis coordinately modulated the virulence-associated interactive metabolome of uropathogenic Escherichia coli and human urine.

    Science.gov (United States)

    Su, Qiao; Guan, Tianbing; Lv, Haitao

    2016-04-14

    Uropathogenic Escherichia coli (UPEC) growth in women's bladders during urinary tract infection (UTI) incurs substantial chemical exchange, termed the "interactive metabolome", which primarily accounts for the metabolic costs (utilized metabolome) and metabolic donations (excreted metabolome) between UPEC and human urine. Here, we attempted to identify the individualized interactive metabolome between UPEC and human urine. We were able to distinguish UPEC from non-UPEC by employing a combination of metabolomics and genetics. Our results revealed that the interactive metabolome between UPEC and human urine was markedly different from that between non-UPEC and human urine, and that UPEC triggered much stronger perturbations in the interactive metabolome in human urine. Furthermore, siderophore biosynthesis coordinately modulated the individualized interactive metabolome, which we found to be a critical component of UPEC virulence. The individualized virulence-associated interactive metabolome contained 31 different metabolites and 17 central metabolic pathways that were annotated to host these different metabolites, including energetic metabolism, amino acid metabolism, and gut microbe metabolism. Changes in the activities of these pathways mechanistically pinpointed the virulent capability of siderophore biosynthesis. Together, our findings provide novel insights into UPEC virulence, and we propose that siderophores are potential targets for further discovery of drugs to treat UPEC-induced UTI.

  20. Generating a non-integrating human induced pluripotent stem cell bank from urine-derived cells.

    Directory of Open Access Journals (Sweden)

    Yanting Xue

    Full Text Available Induced pluripotent stem cell (iPS cell holds great potential for applications in regenerative medicine, drug discovery, and disease modeling. We describe here a practical method to generate human iPS cells from urine-derived cells (UCs under feeder-free, virus-free, serum-free condition and without oncogene c-MYC. We showed that this approach could be applied in a large population with different genetic backgrounds. UCs are easily accessible and exhibit high reprogramming efficiency, offering advantages over other cell types used for the purpose of iPS generation. Using the approach described in this study, we have generated 93 iPS cell lines from 20 donors with diverse genetic backgrounds. The non-viral iPS cell bank with these cell lines provides a valuable resource for iPS cells research, facilitating future applications of human iPS cells.

  1. Nitrogen recovery from source-separated human urine using clinoptilolite and preliminary results of its use as fertilizer.

    Science.gov (United States)

    Beler-Baykal, B; Allar, A D; Bayram, S

    2011-01-01

    The use of source separated human urine as fertilizer is one of the major suggestions of the new sanitation concept ECOSAN. Urine is rich in nitrogen, phosphorus and potassium which act as plant nutrients, however its salinity is high for agricultural and landscape purposes. Moreover, characteristics change significantly throughout storage where salinity increases to higher values as the predominant form of nitrogen shifts from urea to ammonium. Transferring nitrogen in human urine onto the natural zeolite clinoptilolite and using the subsequently recovered ammonium from the exhausted clinoptilolite for agricultural/landscape purposes is suggested as an indirect route of using urine in this work. Results reporting the outcome of the proposed process together with characterization of fresh and stored urine, and preliminary work on the application of the product on the landscape plant Ficus elastica are presented. Up to 97% of the ammonium in stored urine could be transferred onto clinoptilolite through ion exchange and about 88% could be recovered subsequently from exhausted clinoptilolite, giving an overall recovery of 86%. Another important merit of the suggested process was the successful elimination of salinity. Preliminary experiments with Ficus elastica had shown that the product, i.e. clinoptilolite exhausted with ammonium, was compatible with the synthetic fertilizer tested.

  2. [Analysis of protein ratios based on the studies of human urine proteome in an experiment with 105-day isolation].

    Science.gov (United States)

    2012-01-01

    Purpose of the work was to study urine proteome of healthy human subjects during 105-day isolation in controlled environment of the IBMP chamber with a self-sustained life support system. The parameters under study were diurnal rhythm, physical activity, water intake, as well as consumption of sodium, protein and some other basic nutrients. Urine was sampled by 6 male subjects at the age of 25 to 40 years admitted to the experiment by the medical certification commission. The studies were performed with the use of proteome data acquisition technologies; the cutting-edge bio-information analysis techniques including reconstruction of associative gene networks were applied to investigate the urine proteome profile, to structure experimental data in light of the existing physiological concepts and to formulate viable hypotheses for ensuing experimental verification. Proteins of different origin were identified in urine; appearance or disappearance of proteins was in tight correlation with salt consumption.

  3. Determination of salbutamol and salbutamol glucuronide in human urine by means of liquid chromatography-tandem mass spectrometry

    DEFF Research Database (Denmark)

    Mareck, Ute; Guddat, Sven; Schwenke, Anne;

    2012-01-01

    The determination of salbutamol and its glucuronide in human urine following the inhalative and oral administration of therapeutic doses of salbutamol preparations was performed by means of direct urine injection utilizing liquid chromatography-tandem mass spectrometry (LC-MS/MS) and employing d(3......)-salbutamol and d(3)-salbutamol glucuronide as internal standards. Unconjugated salbutamol was detected in all administration study urine samples. Salbutamol concentrations following inhalation were commonly (99%) below 1000 ng/ml whereas values after oral administration frequently (48%) exceeded...... this threshold. While salbutamol glucuronide was not detected in urine samples collected after inhalation of the drug, 26 out of 82 specimens obtained after oral application contained salbutamol glucuronide with a peak value of 63 ng/ml. The percentage of salbutamol glucuronide compared to unconjugated...

  4. Improvements in culturing exfoliated urothelial cells in vitro from human urine.

    Science.gov (United States)

    Belik, Rouslana; Follmann, Wolfram; Degen, Gisela H; Roos, Peter H; Blaszkewicz, Meinolf; Knopf, H Jurgen; Golka, Klaus

    2008-01-01

    Human bladder cancer is a common malignant tumor that may be produced by factors such as lifestyle, environment and occupation. The aim of this study was to evaluate parameters related to the viability of exfoliated urothelial cells. Exfoliated urothelial cells were obtained from 83 urine samples of 22 healthy participants (20-53 yr). From 67 of these samples, cells were transferred to collagen-coated 24-well plates. Parameters including sample volume, pH, osmolality and participant age and gender were examined on cell viability. In successive cultures, the numbers of cell colonies and cells per cell colony were determined. The number of viable cells in the urinary sediments of males varied from 0 to 6.5 x 10(3) cells per sample (mean 1 x 10(3)). Higher cell numbers in urine samples from females (6 x 10(3)) were due to considerable amounts of exfoliated vaginal cells. Cell numbers in males were positively related to volume, osmolality, and pH of the samples, as well as to the retention time of urine in the bladder. Cell proliferation was achieved in 25 out of 67 samples and was positively related to sample osmolality and pH. Participant age and content of urinary oxalates exerted negative effects on cell proliferation in vitro. The mean number of cell colonies per sample was 1.7. The mean cell number per colony was 11.7 x 10(3). It appears that high variability in individual excretion of urothelial cells able to proliferate is a limiting factor for routine use of these cells for in vitro toxicology.

  5. Chinese population exposure to triclosan and triclocarban as measured via human urine and nails.

    Science.gov (United States)

    Yin, Jie; Wei, Ling; Shi, Ying; Zhang, Jing; Wu, Qingqing; Shao, Bing

    2016-10-01

    Triclosan (TCS) and triclocarban (TCC) exposures are highly concerned due to their suspected endocrine-disrupting effects. The present study investigated TCS and TCC exposure levels in the general Chinese population by biomonitoring human urine and nail samples. TCS (69-80 %) and TCC (99-100 %) were frequently detected, which demonstrates that the general Chinese population has extensive exposure to these chemicals. The geometric mean (GM) urinary concentrations were 0.40 μg/g creatinine (creat), 95 % confidence interval (CI) 0.30-0.56, for TCS and 0.40 μg/g creat, 95 % CI 0.29-0.56, for TCC. On the other hand, the GM levels of TCS and TCC were 13.57 (5.67 μg/kg) and 84.66 μg/kg (41.50 μg/kg) in fingernail (toenail) samples, respectively, indicating that the levels in fingernails were approximately twice as high as those in toenails. Pearson's correlation coefficients between the urine and fingernail (toenail) samples were 0.715 (0.614) for TCS and 0.829 (0.812) for TCC. These data suggest that nail samples can be applied to the biomonitoring for TCS and TCC in the general population. We observed that the levels of both chemicals were higher in females than in males for urine and fingernail samples, but no significant differences were found between different genders for either compound in toenails. Nineteen- to 29-year-olds had the highest TCS levels in their nail samples, whereas TCC levels did not differ with regard to age. Region of residence significantly influenced TCS and TCC concentrations in the three biological matrices measured.

  6. Bilirubin - urine

    Science.gov (United States)

    Conjugated bilirubin - urine; Direct bilirubin - urine ... Bilirubin is not normally found in the urine. ... Increased levels of bilirubin in the urine may be due to: Biliary tract disease Cirrhosis Gallstones in the biliary tract Hepatitis Liver disease ...

  7. Survival of enteric bacteria in source-separated human urine used ...

    African Journals Online (AJOL)

    MAKAYA

    the quarter-diluted urine at 25°C. Survival times of the tested bacteria were more shortened in ammonia ... the urine treatment by storage may preserve the useful nutrients ... MATERIALS AND METHODS .... Microsoft Excel package was used.

  8. Determination of Ephedrine Alkaloids in Human Urine and Plasma by Liquid Chromatography/Tandem Mass Spectrometry: Collaborative Study

    Science.gov (United States)

    Trujillo, William A.; Sorenson, Wendy R.

    2008-01-01

    A collaborative study was conducted to evaluate the accuracy and precision of a method for ephedrine-type alkaloids (i.e., norephedrine, norpseudoephedrine, ephedrine, pseudoephedrine, methylephedrine, and methylpseudoephedrine) in human urine and plasma. The amount of ephedrine-type alkaloids present was determined using liquid chromatography (LC) with tandem mass selective detection. The test samples were diluted to reflect a concentration of 5.00–100 ng/mL for each alkaloid. An internal standard was added and the alkaloids were separated using a 5 μm phenyl LC column with an ammonium acetate, glacial acetic acid, acetonitrile, and water mobile phase. Eight blind duplicates of human urine and eight blind duplicates of human plasma were analyzed by 10 collaborators. In addition to negative controls, test portions of urine and plasma were fortified at 3 different levels with each of the 6 ephedrine-type alkaloids at approximately 1, 2, and 5 μg/mL for urine and 100, 200, and 500 ng/mL for plasma. On the basis of the accuracy and precision results for this collaborative study, it is recommended that this method be adopted Official First Action for the determination of 6 different ephedrine-type alkaloids in human urine and plasma. PMID:14509420

  9. Captan metabolism in humans yields two biomarkers, tetrahydrophthalimide (THPI) and thiazolidine-2-thione-4-carboxylic acid (TTCA) in urine.

    Science.gov (United States)

    Krieger, R I; Thongsinthusak, T

    1993-01-01

    Captan fungicide (N-(trichloromethylthio)-4-cyclohexene-1,2-dicarboximide) metabolism in two human volunteers rapidly yields THPI (tetrahydrophthalimide) and TTCA (thiazolidine-2-thione-4-carboxylic acid). The work was done to evaluate usefulness of TTCA and THPI as biomarkers of occupational exposure and to compare human and rat dermal absorption and metabolism. THPI in 12h urine ranged from MDL (5 ppb) to 640 ppb and was stable for at least one year. TTCA was also a stable metabolite, but the MDL was 50 ppb. THPI was detectable in urine for 72 hours following oral dosages of 1 mg/kg, but most was eliminated 0-24 h. No THPI was detectable in urine following application of a chloroform solution to hands, forearms, or inguinal region. Dermal absorption and metabolism of captan are substantially different in humans and rats.

  10. Selenium speciation in pretreated human urine by ion-exchange chromatography and ICP-MS detection

    DEFF Research Database (Denmark)

    Gammelgaard, Bente; Jons, O.; Bendahl, L.

    2001-01-01

    Urine samples were extracted by benzo-15-crown-5-ether to remove sodium and potassium. More than 90% of the sodium and potassium content of the urine was removed with this extraction. In a cation-exchange system based on oxalic acid at pH 3, chromatography of an untreated urine pool resulted in a...

  11. Determination of raloxifene hydrochloride in human urine by LC-MS-MS

    Directory of Open Access Journals (Sweden)

    K. Tharpa

    2009-09-01

    Full Text Available A sensitive and selective liquid chromatographic-tandem mass spectrometric (LC-MS-MS method was developed to determine raloxifene hydrochloride (RLX in human urine. After a solid-phase extraction with SPE cartridge, the urine sample was analyzed on a C18 column (Symmetry 3.5μm; 50 mm4.6 mm i.d interfaced with a triple quadruple tandem mass spectrometer. A positive electrospray ionization was employed as the ionization source. The mobile phase consisted of ammonium acetate (pH 4.0–acetonitrile (60:40, v/v.The method was linear over a concentration range of 20–1000 ng mL-1. The lower limit of quantitation was 20 ng mL-1. The intra-day and inter-day relative standard deviation across three validation runs over the entire concentration range was <10.5%. The accuracy determined at three concentrations (50, 500 and 850 ng mL-1 RLX was within ±0.84% in terms of relative errors.

  12. Characterization and quantification of endogenous fatty acid nitroalkene metabolites in human urine.

    Science.gov (United States)

    Salvatore, Sonia R; Vitturi, Dario A; Baker, Paul R S; Bonacci, Gustavo; Koenitzer, Jeffrey R; Woodcock, Steven R; Freeman, Bruce A; Schopfer, Francisco J

    2013-07-01

    The oxidation and nitration of unsaturated fatty acids transforms cell membrane and lipoprotein constituents into mediators that regulate signal transduction. The formation of 9-NO2-octadeca-9,11-dienoic acid and 12-NO2-octadeca-9,11-dienoic acid stems from peroxynitrite- and myeloperoxidase-derived nitrogen dioxide reactions as well as secondary to nitrite disproportionation under the acidic conditions of digestion. Broad anti-inflammatory and tissue-protective responses are mediated by nitro-fatty acids. It is now shown that electrophilic fatty acid nitroalkenes are present in the urine of healthy human volunteers (9.9 ± 4.0 pmol/mg creatinine); along with electrophilic 16- and 14-carbon nitroalkenyl β-oxidation metabolites. High resolution mass determinations and coelution with isotopically-labeled metabolites support renal excretion of cysteine-nitroalkene conjugates. These products of Michael addition are in equilibrium with the free nitroalkene pool in urine and are displaced by thiol reaction with mercury chloride. This reaction increases the level of free nitroalkene fraction >10-fold and displays a K(D) of 7.5 × 10(-6) M. In aggregate, the data indicates that formation of Michael adducts by electrophilic fatty acids is favored under biological conditions and that reversal of these addition reactions is critical for detecting both parent nitroalkenes and their metabolites. The measurement of this class of mediators can constitute a sensitive noninvasive index of metabolic and inflammatory status.

  13. Determination of amphetamines in human urine by headspace solid-phase microextraction and gas chromatography.

    Science.gov (United States)

    Raikos, Nikolaos; Christopoulou, Klio; Theodoridis, Georgios; Tsoukali, Heleni; Psaroulis, Dimitrios

    2003-06-05

    Solid-phase microextraction (SPME) is under investigation for its usefulness in the determination of a widening variety of volatile and semivolatile analytes in biological fluids and materials. Semivolatiles are increasingly under study as analytical targets, and difficulties with small partition coefficients and long equilibration times have been identified. Amphetamines were selected as semivolatiles exhibiting these limitations and methods to optimize their determination were investigated. A 100- micro m polydimethylsiloxane (PDMS)-coated SPME fiber was used for the extraction of the amphetamines from human urine. Amphetamine determination was made using gas chromatography (GC) with flame-ionization detection (FID). Temperature, time and salt saturation were optimized to obtain consistent extraction. A simple procedure for the analysis of amphetamine (AMP) and methamphetamine (MA) in urine was developed and another for 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxy-N-methamphetamine (MDMA) and 3,4-methylenedioxy-N-ethylamphetamine (MDEA) using headspace solid-phase microextraction (HS-SPME) and GC-FID. Higher recoveries were obtained for amphetamine (19.5-47%) and methamphetamine (20-38.1%) than MDA (5.1-6.6%), MDMA (7-9.6%) and MDEA (5.4-9.6%).

  14. Nitrous oxide emissions from cattle excreta applied to a Scottish grassland: effects of soil and climatic conditions and a nitrification inhibitor.

    Science.gov (United States)

    Bell, M J; Rees, R M; Cloy, J M; Topp, C F E; Bagnall, A; Chadwick, D R

    2015-03-01

    Dung and urine excreted onto grasslands are a major source of nitrous oxide (N2O). These N2O emissions stem from inefficient utilisation of nitrogen (N) ingested by ruminants, and the inability of pasture to utilise the deposited N. Predicted growth in dairy and meat consumption means that there is a requirement to quantify N2O emissions, and investigate emission reduction mechanisms. Three 12 month 'seasonal' experiments were undertaken at Crichton, SW Scotland, where N2O emissions were measured from applications of cattle urine, dung, artificial urine and urine+a nitrification inhibitor (NI), dicyandiamide (DCD). The three application timings were 'spring', 'summer' and 'autumn', representative of early-, mid- and late grazing seasons. N2O emissions were measured from static chambers for 12 months. The aim was to quantify emissions from cattle excreta, and determine their dependence on the season of application, and the respective contribution of dung and urine to total excreta emissions. Measurement from NI amended urine was made to assess DCD's potential as an emission mitigation tool. Emissions were compared to the IPCC's default emission factor (EF) of 2% for cattle excreted N. Mean annual cumulative emissions from urine were the highest when applied in summer (5034 g N2O-N ha(-1)), with lower emissions from spring (1903 g N2O-N ha(-1)) and autumn (2014 g N2O-N ha(-1)) application, most likely due to higher temperatures and soil moisture conducive to both nitrification and denitrification in the summer months. Calculated EFs were significantly greater from urine (1.1%) than dung (0.2%) when excreta was applied in summer, and EFs varied with season of application, but in all experiments were lower than the IPCC default of 2%. These results support both lowering and disaggregating this EF into individual EFs for dung and urine. Addition of DCD to urine caused no significant reduction in emissions, suggesting that more research is required into its use as a

  15. Further biochemical characterization of an Na+ pump inhibitor purified from human urine.

    Science.gov (United States)

    Crabos, M; Grichois, M L; Guicheney, P; Wainer, I W; Cloix, J F

    1987-01-02

    An increase in endogenous Na+,K+-ATPase inhibitor(s) with digitalis-like properties has been reported in chronic renal insufficiency, in Na+-dependent experimental hypertension and in some essential hypertensive patients. The present study specifies some properties and some biochemical characteristics of a semipurified compound from human urine having digitalis-like properties. The urine-derived inhibitor (endalin) inhibits Na+,K+-ATPase activity and [3H]-ouabain binding, and cross-reacts with anti-digoxin antibodies. The inhibitory effect on ATPases of endalin is higher on Na+,K+-ATPase than on Mg2+-ATPase and Ca2+-ATPase. The mechanism of endalin action on highly purified Na+,K+-ATPase was compared to that of ouabain and was similar in that it reversibly inhibited Na+,K+-ATPase activity; it inhibited Na+,K+-ATPase non-competitively with ATP; its inhibitory effect was facilitated by Na+; K+ decreased its inhibitory effect on Na+,K+-ATPase; it competitively inhibited ouabain binding to the enzyme; its binding was maximal in the presence of Mg2+ and Pi; it decreased the Na+ pump activity in human erythrocytes; it reduced serotonin uptake by human platelets; and it was diuretic and natriuretic in rat bioassay. The endalin differed from ouabain in only three aspects: its inhibitory effect was not really specific for Na+,K+-ATPase; its binding to the enzyme was undetectable in the presence of Mg2+ and ATP; it was not kaliuretic in rat bioassay. Endalin is a reversible and partial specific inhibitor of Na+,K+-ATPase, its Na+,K+-ATPase inhibition closely resembles that of ouabain and it could be considered as one of the natriuretic hormones.

  16. [Determination of phenobarbital in human urine and serum using flow injection chemiluminescence].

    Science.gov (United States)

    Li, X; Niu, L C; He, X L; Song, Z H

    2012-01-01

    A sensitive chemiluminescence method, based on the enhancive effect of phenobarbital on the chemiluminescence reaction between luminol and dissolved oxygen in a flow injection system, was proposed for the determination of phenobarbital. The chemiluminescence intensity responded to the concentration of phenobarbital linearly ranging from 0.05 to 10 ng x ml(-1) with the detection limit of 0.02 ng x ml(-1) (3 sigma). At a flow rate of 2.0 ml x min(-1), a complete determination of phenobarbital, including sampling and washing, could be accomplished in 0.5 min, offering the sampling efficiency of 120 h(-1) accordingly. The method was applied successfully in an assay of PB for pharmaceutical preparations, human urine and serum without any pretreatment with recovery from 95.7 to 106.7% and RSDs of less than 3.0%.

  17. Determination of uric acid in human urine by ion chromatography with conductivity detector

    Institute of Scientific and Technical Information of China (English)

    Fu Yong Zhao; Zong Hua Wang; Hui Wang; Rui Zhao; Ming Yu Ding

    2011-01-01

    A simple, fast, precise and eco-friendly analytical method for the determination of uric acid (UA) in human urine by ion chromatography (IC) was established. The sample pretreatment was not required, only needed centrifugation and filtration. The separation was carried out on a cation exchange column with 2.0 mmol/L nitric acid as mobile phase at the flow-rate 1.0 mL/min. A non-suppressed conductivity detector was used. The IC analysis time for one run was within 10 min under the optimized IC condition. The detection limits were 0.5 μg/L(S/N = 3) for uric acid. The recovery was 100.1 % while the relative standard deviation (RSD) was 1.8% from 10 measurements.

  18. Adsorptive Cathodic Stripping Voltammetric Determination of Cefoperazone in Bulk Powder, Pharmaceutical Dosage Forms, and Human Urine

    Science.gov (United States)

    Hoang, Vu Dang; Huyen, Dao Thi; Phuc, Phan Hong

    2013-01-01

    The electroreduction behaviour and determination of cefoperazone using a hanging mercury drop electrode were investigated. Cyclic voltammograms of cefoperazone recorded in universal Britton-Robinson buffers pH 3–6 exhibited a single irreversible cathodic peak. The process was adsorption-controlled. Britton-Robinson buffer 0.04 M pH 4.0 was selected as a supporting electrolyte for quantitative purposes by differential pulse and square wave adsorptive cathodic stripping voltammetry. The experimental voltammetric conditions were optimized using Central Composite Face design. A reduction wave was seen in the range from −0.7 to −0.8 V. These voltammetric techniques were successfully validated as per ICH guidelines and applied for the determination of cefoperazone in its single and sulbactam containing powders for injection and statistically comparable to USP-HPLC. They were further extended to determine cefoperazone in spiked human urine with no matrix effect. PMID:24109542

  19. Adsorptive Cathodic Stripping Voltammetric Determination of Cefoperazone in Bulk Powder, Pharmaceutical Dosage Forms, and Human Urine

    Directory of Open Access Journals (Sweden)

    Vu Dang Hoang

    2013-01-01

    Full Text Available The electroreduction behaviour and determination of cefoperazone using a hanging mercury drop electrode were investigated. Cyclic voltammograms of cefoperazone recorded in universal Britton-Robinson buffers pH 3–6 exhibited a single irreversible cathodic peak. The process was adsorption-controlled. Britton-Robinson buffer 0.04 M pH 4.0 was selected as a supporting electrolyte for quantitative purposes by differential pulse and square wave adsorptive cathodic stripping voltammetry. The experimental voltammetric conditions were optimized using Central Composite Face design. A reduction wave was seen in the range from −0.7 to −0.8 V. These voltammetric techniques were successfully validated as per ICH guidelines and applied for the determination of cefoperazone in its single and sulbactam containing powders for injection and statistically comparable to USP-HPLC. They were further extended to determine cefoperazone in spiked human urine with no matrix effect.

  20. Adsorptive cathodic stripping voltammetric determination of cefoperazone in bulk powder, pharmaceutical dosage forms, and human urine.

    Science.gov (United States)

    Hoang, Vu Dang; Huyen, Dao Thi; Phuc, Phan Hong

    2013-01-01

    The electroreduction behaviour and determination of cefoperazone using a hanging mercury drop electrode were investigated. Cyclic voltammograms of cefoperazone recorded in universal Britton-Robinson buffers pH 3-6 exhibited a single irreversible cathodic peak. The process was adsorption-controlled. Britton-Robinson buffer 0.04 M pH 4.0 was selected as a supporting electrolyte for quantitative purposes by differential pulse and square wave adsorptive cathodic stripping voltammetry. The experimental voltammetric conditions were optimized using Central Composite Face design. A reduction wave was seen in the range from -0.7 to -0.8 V. These voltammetric techniques were successfully validated as per ICH guidelines and applied for the determination of cefoperazone in its single and sulbactam containing powders for injection and statistically comparable to USP-HPLC. They were further extended to determine cefoperazone in spiked human urine with no matrix effect.

  1. Urine-derived induced pluripotent stem cells as a modeling tool to study rare human diseases.

    Science.gov (United States)

    Shi, Liang; Cui, Yazhou; Luan, Jing; Zhou, Xiaoyan; Han, Jinxiang

    2016-08-01

    Rare diseases with a low prevalence are a key public health issue because the causes of those diseases are difficult to determine and those diseases lack a clearly established or curative treatment. Thus, investigating the molecular mechanisms that underlie the pathology of rare diseases and facilitating the development of novel therapies using disease models is crucial. Human induced pluripotent stem cells (iPSCs) are well suited to modeling rare diseases since they have the capacity for self-renewal and pluripotency. In addition, iPSC technology provides a valuable tool to generate patient-specific iPSCs. These cells can be differentiated into cell types that have been affected by a disease. These cells would circumvent ethical concerns and avoid immunological rejection, so they could be used in cell replacement therapy or regenerative medicine. To date, human iPSCs could have been generated from multiple donor sources, such as skin, adipose tissue, and peripheral blood. However, these cells are obtained via invasive procedures. In contrast, several groups of researchers have found that urine may be a better source for producing iPSCs from normal individuals or patients. This review discusses urinary iPSC (UiPSC) as a candidate for modeling rare diseases. Cells obtained from urine have overwhelming advantages compared to other donor sources since they are safely, affordably, and frequently obtained and they are readily obtained from patients. The use of iPSC-based models is also discussed. UiPSCs may prove to be a key means of modeling rare diseases and they may facilitate the treatment of those diseases in the future.

  2. Direct measurement of the glucuronide conjugate of 1-hydroxypyrene in human urine by using liquid chromatography with tandem mass spectrometry.

    Science.gov (United States)

    Kakimoto, Kensaku; Toriba, Akira; Ohno, Takanori; Ueno, Mariko; Kameda, Takayuki; Tang, Ning; Hayakawa, Kazuichi

    2008-05-15

    To evaluate human exposure to polycyclic aromatic hydrocarbons (PAHs), we developed a rapid, simple and sensitive method for determining 1-hydroxypyrene-glucuronide (1-OHP-G) in human urine. To improve precision, a deuterated glucuronide was used as an internal standard. The method requires only 1 mL of urine. The urine was treated with a mixed-mode anion-exchange and reversed-phase solid-phase extraction cartridge (Oasis MAX). The analytes were analyzed with a C(18) reversed-phase column with a gradient elution, followed by tandem mass spectrometry with electrospray ionization in negative ion mode. The detection limit of 1-OHP-G (corresponding to a signal-to-noise ratio of 3) was 0.13 fmol/injection. Urinary concentrations of 1-OHP-G determined by this method were strongly correlated (r(2)=0.961) with concentrations of 1-hydroxypyrene by conventional HPLC with fluorescence detection.

  3. Quantitation of Metformin in Human Plasma and Urine by Hydrophilic Interaction Liquid Chromatography and Application to a Pharmacokinetic Study

    DEFF Research Database (Denmark)

    Nielsen, Flemming; Hougaard Christensen, Mette Marie; Brøsen, Kim

    2014-01-01

    : We describe an analytical method for the quantification of the widely used antihyperglycemic agent, metformin, in human plasma and urine. The separation was performed using isocratic hydrophilic interaction liquid chromatography on a Luna hydrophilic interaction liquid chromatography column (125.......5% in plasma and 1.6% to 6.2% in urine. Between-day reproducibility ranged from 2.9% to 5.3% in plasma and 0.6% to 1.8% in urine. The inaccuracy expressed as bias ranged from -3.1% to 1.9% in plasma and from -7.2% to 0.7% in urine. The lower limit of quantification for metformin in plasma was 5 ng....../mL and in urine was 40 ng/mL. The method was therefore considered to be precise, accurate, reproducible, and sensitive enough to be appropriate for pharmacokinetic studies of metformin. The applicability of the method for human pharmacokinetic studies was demonstrated by dosing a healthy male volunteer with 500...

  4. Semi-quantitative tests of cyanide in foods and excreta of Three Hapalemur species in Madagascar.

    Science.gov (United States)

    Yamashita, Nayuta; Tan, Chia L; Vinyard, Christopher J; Williams, Cathy

    2010-01-01

    Three sympatric Hapalemur species (H. g. griseus, H. aureus, and H. (Prolemur) simus) in Ranomafana National Park, Madagascar are known to eat bamboo food parts that contain cyanide. How these lemurs avoid cyanide poisoning remains unknown. In this study, we tested for the presence/absence of cyanide in bamboo lemur foods and excreta to (1) document patterns of cyanide consumption among species with respect to diet, (2) identify routes of elimination of cyanide from the gastrointestinal tract, and (3) determine whether cyanide is absorbed from the diet. We tested 102 food, urine, and fecal samples for hydrogen cyanide (HCN) during two "pre-dry" seasons (April 2006, May 2007) using commercially available Cyantesmo test strips. The test strips changed color in the presence of HCN, and we recorded color change on a scale of 0 (no change) to 5 (cobalt) at preset intervals with a final score taken at 24 hr. We detected cyanide in bamboo food parts and urine of all three Hapalemur species. Time to color change of the test strips ranged from almost instantaneous to >12 hr incubation. Of the foods tested, only bamboo contained cyanide, but results differed among bamboo species and plant parts of the same species. Specifically, branch shoot and culm pith of the giant bamboo produced strong, immediate reactions to the test paper, whereas parts of liana bamboos produced either weak or no color change. Cyanide was present in almost all urine samples but rarely in fecal samples. This suggests that dietary cyanide is absorbed in the gastrointestinal tract of the Hapalemur species and excreted, at least in part, by the kidneys. Samples from H. griseus exhibited lower, though still detectable, cyanide levels compared with H. simus and H. aureus. Differences among lemur species appear to be related to the specific bamboo parts consumed.

  5. Survey of attitudes and perceptions of urine-diverting toilets and human waste recycling in Hawaii

    Energy Technology Data Exchange (ETDEWEB)

    Lamichhane, Krishna M., E-mail: lamichha@hawaii.edu [University of Hawaii, Department of Civil and Environmental Engineering, 2540 Dole Street, Holmes Hall 283, Honolulu, Hawaii 96822 (United States); Babcock, Roger W., E-mail: rbabcock@hawaii.edu [University of Hawaii, Department of Civil and Environmental Engineering, Holmes Hall 383, Honolulu, Hawaii 96822 (United States)

    2013-01-15

    Urine constitutes only about 1% of domestic sewage but contains 50% or more of the excreted nutrients and chemicals like hormones and pharmaceutical residues. Urine diverting toilet (UDT) systems can be considered a more sustainable alternative to wastewater management because they allow nutrient recycling, reduce water use, and allow source-separation of hormones and chemicals that can harm the environment. An online survey was conducted to determine whether UDTs are acceptable to the general public in Hawaii and if attitudes and perceptions towards it and human waste (HW) recycling vary with age, sex, level of education, religious affiliation, ethnicity, and employment status. The survey was also intended to detect possible drivers and barriers for the UDTs. Variations on variables were tested at 5% significance (p = 0.05) level (Chi-squared test or ANOVA) and considered significantly different if the p-value was less than 0.05. The results were encouraging as more than 60% are willing to pay extra for the UDT, while only 22% knew that such systems existed. No statistically significant difference was found between males and females on all survey questions at the 5% level. However, females had higher willingness to pay (WTP) than males and WTP increased with age and income. The WTP of Caucasians was higher than Asians and differed significantly. Some respondents expressed concern about the legal provisions for recycling of HW. The survey results indicate that with a public education program, it is possible that most people would be willing to adopt UDTs and HW recycling with incurred societal benefits of reduced water and fertilizer use, reduced greenhouse gas emissions, and collection of micropollutants at the source to prevent their entry into waterways. Because of the small sample size (N = 132, 13% response rate) the survey is not representative but may be indicative of the general attitude of Hawaiian people. - Highlights: ► Urine diverting toilets (UDTs

  6. Selenium speciation in human urine samples by LC- and CE-ICP-MS-separation and identification of selenosugars

    DEFF Research Database (Denmark)

    Gammelgaard, Bente; Bendahl, L.

    2004-01-01

    Human urine samples were analysed by a reversed-phase chromatographic system and an ion-pair chromatographic system. The chromatographic system, was connected to the ICP-MS either by a microconcentric nebulizer (MCN) in combination with a cyclonic spraychamber or by a modified direct injection...

  7. Simultaneous kinetic spectrophotometric determination of norfloxacin and rifampicin in pharmaceutical formulation and human urine samples by use of chemometrics approaches

    Institute of Scientific and Technical Information of China (English)

    KOKOT; Serge

    2008-01-01

    A kinetic spectrophotometric method with aid of chemometrics is proposed for the simultaneous determination of norfloxacin and rifampicin in mixtures. The proposed method was applied for the simultaneous determination of these two compounds in pharmaceutical formulation and human urine samples,and the results obtained are similar to those obtained by high performance liquid chromatography.

  8. Detection and sequencing of West Nile virus RNA from human urine and serum samples during the 2014 seasonal period.

    Science.gov (United States)

    Nagy, Anna; Bán, Enikő; Nagy, Orsolya; Ferenczi, Emőke; Farkas, Ágnes; Bányai, Krisztián; Farkas, Szilvia; Takács, Mária

    2016-07-01

    West Nile virus, a widely distributed mosquito-borne flavivirus, is responsible for numerous animal and human infections in Europe, Africa and the Americas. In Hungary, the average number of human infections falls between 10 and 20 cases each year. The severity of clinically manifesting infections varies widely from the milder form of West Nile fever to West Nile neuroinvasive disease (WNND). In routine laboratory diagnosis of human West Nile virus infections, serological methods are mainly applied due to the limited duration of viremia. However, recent studies suggest that detection of West Nile virus RNA in urine samples may be useful as a molecular diagnostic test for these infections. The Hungarian National Reference Laboratory for Viral Zoonoses serologically confirmed eleven acute human infections during the 2014 seasonal period. In three patients with neurological symptoms, viral RNA was detected from both urine and serum specimens, albeit for a longer period and in higher copy numbers with urine. Phylogenetic analysis of the NS3 genomic region of three strains and the complete genome of one selected strain demonstrated that all three patients had lineage-2 West Nile virus infections. Our findings reaffirm the utility of viral RNA detection in urine as a molecular diagnostic procedure for diagnosis of West Nile virus infections.

  9. [Simultaneous determination of four mercapturic acids in human urine using solid phase extraction and liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Hou, Hongwei; Xiong, Wei; Gao, Na; Song, Dongkui; Tang, Gangling; Hu, Qingyuan

    2011-01-01

    A method was developed for the simultaneous extraction and determination of four mercapturic acids (MAs), N-acetyl-S-(3,4-dihydroxybutyl)-L-cysteine (DHBMA), N-acetyl-S-(3-hydroxypropyl)-L-cysteine (3-HPMA), N-acetyl-S-(2-carboxyethyl)-L-cysteine ( CEMA) and S-phenylmercapturic acid (SPMA), in human urine using solid phase extraction and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Frozen urine samples were thawed at room temperature, and centrifuged to remove any settled precipitate. The supernatant was then purified and concentrated by a C18 solid phase extraction column, and analyzed by HPLC-MS/MS in the multiple reaction monitoring (MRM) mode for the quantitative analysis. The ranges of recovery for DHBMA, 3-HPMA, CEMA and SPMA spiked in human urine matrix at three concentration levels were 105.6%-124.4%, 102.7%-106.5%, 103.2%-103.9% and 101.7%-104.3%, respectively, with the relative standard deviations of 2.6%-7.7%. The limits of detection (LOD, S/N > or = 3) were 0. 062, 0. 031, 0. 020 and 0. 003 microg/L for DHBMA, 3-HPMA, CEMA and SPMA, respectively. The method was successfully used to detect 4 MAs in 37 human urine samples from smokers and non-smokers. It was found that the contents of 3-HPMA, CEMA and SPMA in the urines from cigarette smokers were about three to six-fold more than those in the urines from the non-smokers.

  10. Toxicokinetics of colchicine in humans: analysis of tissue, plasma and urine data in ten cases.

    Science.gov (United States)

    Rochdi, M; Sabouraud, A; Baud, F J; Bismuth, C; Scherrmann, J M

    1992-11-01

    1. A specific and sensitive radioimmunoassay was used to study the toxicokinetics of colchicine in seven cases of acute human poisoning. Post-mortem tissue concentrations of colchicine were measured in three further cases. Depending on the time of patient admission, two disposition processes could be observed. The first, in three patients, admitted early, showed a bi-exponential plasma colchicine decrease, with distribution half-lives of 30, 45 and 90 min. The second, in four patients, admitted late, showed a mono-exponential decrease. Plasma terminal half-lives ranged from 10.6 to 31.7 h for both groups. 2. Pharmacokinetic analysis of urine colchicine data was performed for two patients. The fraction of unchanged colchicine excreted in urine was about 30%, renal clearance was about 13 l h-1 and three-fold less than total body clearance (39 l h-1). The apparent volume of distribution was 21 l kg-1. 3. Post-mortem tissue analysis showed an ubiquitous colchicine distribution. Colchicine accumulated at high concentrations in the bone marrow (more than 600 ng g-1), testicle (400 ng g-1), spleen (250 ng g-1), kidney (200 ng g-1), lung (200 ng g-1) and heart (95 ng g-1); it was also found in the brain (125 ng g-1). 4. This toxicokinetic study shows that after massive ingestion, the disposition parameters and kinetics of colchicine are not markedly modified from those occurring in healthy volunteers. The absorption process was not delayed and the distribution and elimination half-lives were in the range known to occur with therapeutic doses.

  11. False-negative urine human chorionic gonadotropin in molar pregnancy: " The high-dose hook effect" !

    Directory of Open Access Journals (Sweden)

    Sujata Narendra Datti

    2015-01-01

    Full Text Available Failure to detect pregnancy in the emergency situations can have important consequences. These include missing of ectopic pregnancy (the leading cause of first-trimester pregnancy-related maternal death, administration of medications contraindicated in pregnancy, fetal radiation exposure, and medico legal problems. This in turn has led to the dictum to check for pregnancy in all women of child-bearing age group. Urine pregnancy (human chorionic gonadotropin [hCG] test is the commonly used test to rule out pregnancy and has been reported by Griffey et al. in their study to achieve 100% sensitivity and 99.2% specificity in a clinical setting, resulting in a positive predictive value of 98.3% and a negative predictive value of nearly 100%. However, the sensitivity is influenced not only by the quantity of β hCG but on its variants that vary with different weeks of pregnancy. β hCG is present in several variant forms that change in their concentrations at different stages of pregnancy. In spite of its high sensitivity, in the presence of molar pregnancy that is associated with very high levels of β hCG it fails to detect the antigen (β hCG. This is explained by the phenomenon known as "high-dose hook effect" which further leads to delay in diagnosis and treatment. This can be overcome by dilution of the sample. In such cases, diagnosis will be made by serum β hCG and ultrasound (USG. Here, we present a case of gravida 2 para 1 living 1 with 2΍ months amenorrhea with bleeding p/v and pain abdomen of 20 days duration whose urine β hCG was repeatedly negative and diagnosis was made by serum β hCG and USG.

  12. Identification and quantification of metabolites of the fungicide tebuconazole in human urine.

    Science.gov (United States)

    Mercadante, R; Polledri, E; Scurati, S; Moretto, A; Fustinoni, S

    2014-11-17

    Tebuconazole (TEB) is a fungicide used in agriculture; the objective of this work was to identify and quantify TEB metabolites in human urine. Samples from seven vineyard workers exposed to TEB were submitted to liquid chromatography interfaced with a triple quadrupole mass spectrometer, equipped with an electron spray source, and a linear ion trap to gain a profile of candidate metabolites. Based on the presence of the ion m/z 70 in the MS/MS spectra, which corresponds to protonated triazole (a specific moiety of TEB), and the isotopic pattern of the molecular ions, typical of molecules with one chlorine atom, hydroxyl and carboxyl derivatives of TEB, that is, TEB-OH and TEB-COOH, were identified as major metabolites, both as free molecules and as glucuronide (Glc) conjugates. The mean molar fractions were 0.67, 0.13, 0.13, and 0.07 for TEB-O-Glc, TEB-OH, TEB-COO-Glc, and TEB-COOH. Urine samples were submitted to hydrolysis with β-glucuronidase, and the free compounds were quantified in the presence of deuterated TEB (TEB-d6) as the internal standard (IS), by multiple reaction monitoring (MRM) mode. The assay was linear in the ranges of 0.2-600 μg/L and 0.1-240 μg/L for TEB-OH and TEB-COOH, respectively; precision, accuracy, and the limit of quantification (LOQ) were <3.1%, 98-103%, and 0.3 μg/L for both analytes. An evaluation of matrix effects showed that the use of TEB-d6 controlled these sources of bias. The urinary levels of TEB-OH and TEB-COOH in specimens collected from farmers exposed to TEB ranged from 10 to 473 and from 3 to 159 μg/L, respectively.

  13. Screening and Identification of Mitragynine and 7-Hydroxymitragynine in Human Urine by LC-MS/MS

    Directory of Open Access Journals (Sweden)

    Hanzhuo Fu

    2015-05-01

    Full Text Available Kratom is a tree planted in Southeast Asia, including Thailand, Malaysia, Myanmar (Burma and elsewhere in the region. A long history of usage and abuse of kratom has led to the classification of kratom as a controlled substance in its native Thailand and other Southeast Asian countries. However, kratom is not controlled in the United States, and the wide availability of kratom on the Internet and in the streets has led to its emergence as an herbal drug of misuse. With the increasing popularity of kratom, efficient protocols are needed to detect kratom use. In this study, a rapid method for the analysis of kratom compounds, mitragynine and 7-hydroxymitragynine, in human urine has been developed and validated using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS. The chromatographic system employed a 2.6-μm 100 mm × 2.1 mm phenyl-hexyl analytical column and gradient elution with a 0.4-mL/min flow rate of water and acetonitrile as mobile phases. A triple quadrupole mass spectrometer was used as the detector for data acquisition. The analyst was the quantification software. The established method demonstrated linearity of >0.99 for both analytes, and low detection limits were obtained down to 0.002581 ng/mL for mitragynine and 0.06910 ng/mL for 7-hydroxymitragynine. The validated method has been utilized for clinical analysis of urine for the purpose of mitragynine and 7-hydroxymitragynine detection.

  14. Stored human urine supplemented with wood ash as fertilizer in tomato (Solanum lycopersicum) cultivation and its impacts on fruit yield and quality.

    Science.gov (United States)

    Pradhan, Surendra K; Holopainen, Jarmo K; Heinonen-Tanski, Helvi

    2009-08-26

    This study evaluates the use of human urine and wood ash as fertilizers for tomato cultivation in a greenhouse. Tomatoes were cultivated in pots and treated with 135 kg of N/ha applied as mineral fertilizer, urine + ash, urine only, and control (no fertilization). The urine fertilized plants produced equal amounts of tomato fruits as mineral fertilized plants and 4.2 times more fruits than nonfertilized plants. The levels of lycopene were similar in tomato fruits from all fertilization treatments, but the amount of soluble sugars was lower and Cl(-) was higher in urine + ash fertilized tomato fruits. The beta-carotene content was greater and the NO(3)(-) content was lower in urine fertilized tomato fruits. No enteric indicator microorganisms were detected in any tomato fruits. The results suggest that urine with/without wood ash can be used as a substitute for mineral fertilizer to increase the yields of tomato without posing any microbial or chemical risks.

  15. Pomegranate juice ellagitannin metabolites are present in human plasma and some persist in urine for up to 48 hours.

    Science.gov (United States)

    Seeram, Navindra P; Henning, Susanne M; Zhang, Yanjun; Suchard, Marc; Li, Zhaoping; Heber, David

    2006-10-01

    Ellagitannins (ETs) from pomegranate juice (PJ) are reported to have numerous biological properties, but their absorption and metabolism in humans are poorly understood. To investigate the pharmacokinetics of pomegranate ETs, 18 healthy volunteers were given 180 mL of PJ concentrate, and blood samples were obtained for 6 h afterwards. Twenty-four-hour urine collections were obtained on the day before (-1), the day of (0), and the day after (+1) the study. Ellagic acid (EA) was detected in plasma of all subjects with a maximum concentration of 0.06 +/- 0.01 micromol/L, area under concentration time curve of 0.17 +/- 0.02 (micromol x h) x L(-1), time of maximum concentration of 0.98 +/- 0.06 h, and elimination half-life of 0.71 +/- 0.08 h. EA metabolites, including dimethylellagic acid glucuronide (DMEAG) and hydroxy-6H-benzopyran-6-one derivatives (urolithins), were also detected in plasma and urine in conjugated and free forms. DMEAG was found in the urine obtained from 15 of 18 subjects on d 0, but was not detected on d -1 or +1, demonstrating its potential as a biomarker of intake. Urolithin A-glucuronide was found in urine samples from 11 subjects on d 0 and in the urine from 16 subjects on d +1. Urolithin B-glucuronide was found in the urine of 3 subjects on d 0 and in the urine of 5 subjects on d +1. Urolithins, formed by intestinal bacteria, may contribute to the biological effects of PJ as they may persist in plasma and tissues and account for some of the health benefits noted after chronic PJ consumption. Whether genetic polymorphisms in EA-metabolizing enzymes (e.g., catechol-O-methyl transferase and glucuronosyl transferase) are related to variations in response to PJ remains to be established.

  16. Determination of ephedrine and codeine in human urine by cation-selective exhaustive injection and sweeping micellar electrokinetic chromatography

    Institute of Scientific and Technical Information of China (English)

    Li Jun Li; Si Guang Li; Hai Yan Li; Zhuo Cai; Hao Cheng

    2009-01-01

    A sensitive method for the determination of ephedrine and codeine in human urine by capillary electrophoresis (CE) was described. In order to improve the sensitivity, two online concentration techniques including cation-selective exhaustive injection (CSEI) and sweeping micellar electrokinetic chromatography (sweeping-MEKC) were used. Under the optimum conditions, the detection limits (S/N = 3) were 0.10 μg/L for ephedrine and 0.80 μg/L for codeine. This method was successfully applied to real urine sample analysis.

  17. Direct determination of lead in human urine and serum samples by electrothermal atomic absorption spectrometry and permanent modifiers

    OpenAIRE

    Andrada,Daniel; Pinto,Frederico G.; Magalhães, Cristina Gonçalves; Nunes,Berta R.; Franco,Milton B.; Silva,José Bento Borba da

    2006-01-01

    The object of the present study was the development of alternative methods for the direct determination of lead in undigested samples of human urine and serum by electrothermal atomic absorption spectrometry (ET AAS). Thus, some substances have been investigated to act as chemical modifiers. Volumes of 20 µL of diluted samples, 1 + 1, v/v for urine and 1 + 4, v/v for serum, with HNO3 1% v/v and 0.02% v/v of cetil trimethyl ammonium chloride (CTAC) were prepared directly in the autosampler cup...

  18. Neurotoxic thioether adducts of 3,4-methylenedioxymethamphetamine identified in human urine after ecstasy ingestion.

    Science.gov (United States)

    Perfetti, Ximena; O'Mathúna, Brian; Pizarro, Nieves; Cuyàs, Elisabet; Khymenets, Olha; Almeida, Bruno; Pellegrini, Manuela; Pichini, Simona; Lau, Serrine S; Monks, Terrence J; Farré, Magí; Pascual, Jose Antonio; Joglar, Jesús; de la Torre, Rafael

    2009-07-01

    3,4-Methylenedioxymethamphetamine (MDMA, Ecstasy) is a widely misused synthetic amphetamine derivative and a serotonergic neurotoxicant in animal models and possibly humans. The underlying mechanism of neurotoxicity involves the formation of reactive oxygen species although their source remains unclear. It has been postulated that MDMA-induced neurotoxicity is mediated via the formation of bioreactive metabolites. In particular, the primary catechol metabolites, 3,4-dihydroxymethamphetamine (HHMA) and 3,4-dihydroxyamphetamine (HHA), subsequently cause the formation of glutathione and N-acetylcysteine conjugates, which retain the ability to redox cycle and are serotonergic neurotoxicants in rats. Although the presence of such metabolites has been recently demonstrated in rat brain microdialysate, their formation in humans has not been reported. The present study describes the detection of 5-(N-acetylcystein-S-yl)-3,4-dihydroxymethamphetamine (N-Ac-5-Cys-HHMA) and 5-(N-acetylcystein-S-yl)-3,4-dihydroxyamphetamine (N-Ac-5-Cys-HHA) in human urine of 15 recreational users of MDMA (1.5 mg/kg) in a controlled setting. The results reveal that in the first 4 h after MDMA ingestion approximately 0.002% of the administered dose was recovered as thioether adducts. Genetic polymorphisms in CYP2D6 and catechol-O-methyltransferase expression, the combination of which are major determinants of steady-state levels of HHMA and 4-hydroxy-3-methoxyamphetamine, probably explain the interindividual variability seen in the recovery of N-Ac-5-Cys-HHMA and N-Ac-5-Cys-HHA. In summary, the formation of neurotoxic thioether adducts of MDMA has been demonstrated for the first time in humans. The findings lend weight to the hypothesis that the bioactivation of MDMA to neurotoxic metabolites is a relevant pathway to neurotoxicity in humans.

  19. Urine culture

    Science.gov (United States)

    Culture and sensitivity - urine ... when urinating. You also may have a urine culture after you have been treated for an infection. ... when bacteria or yeast are found in the culture. This likely means that you have a urinary ...

  20. Determination of salbutamol in human plasma and urine using liquid chromatography coupled to tandem mass spectrometry and its pharmacokinetic study.

    Science.gov (United States)

    Zhang, Dujuan; Teng, Yanni; Chen, Keguang; Liu, Sha; Wei, Chunmin; Wang, Benjie; Yuan, Guiyan; Zhang, Rui; Liu, Xiaoyan; Guo, Ruichen

    2012-10-01

    A sensitive and selective liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was developed and validated for the determination of salbutamol in human plasma and urine, and successfully applied to the pharmacokinetic study of salbutamol in Chinese healthy volunteers after inhalation of salbutamol sulfate aerosol. Salbutamol and the internal standard (IS) acetaminophen in plasma and urine were extracted with ethyl acetate, separated on a C(18) reversed-phase column, eluted with mobile phase of acetonitrile-ammonium acetate (5 m m; 30:70, v/v), ionized by positive ion pneumatically assisted electrospray and detected in the multi-reaction monitoring mode using precursor → product ions of m/z 240.2 → 148.1 for salbutamol and 152 → 110 for the IS. The lower limits of quantitation of salbutamol in human plasma and urine by this method were 0.02 and 1 ng/mL, respectively. The specificity, matrix effect, recovery, sensitivity, linearity, accuracy, precision and several stabilities were validated for salbutamol in human plasma and urine. In conclusion, the validation results showed that this method is robust, specific and sensitive, and can successfully fulfill the requirement of clinical pharmacokinetic study of salbutamol in healthy Chinese volunteers.

  1. Trace analysis of sulfamethazine in animal feed, human urine, and wastewater by electron capture gas chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Holder, C.L.; Thompson, H.C. Jr.; Bowman, M.C.

    1981-12-01

    Sulfamethazine, a widely used antibacterial drug additive in feeds for swine, chickens, and cattle, was scheduled for toxicological evaluation because of potential human health hazards associated with its residues in edible animal tissues. Analytical chemical procedures that would ensure proper concentration, homogeneity, and stability of the drug in dosed feed and its safe usage during the animal studies were prerequisites for such toxicological tests. Electron capture gas chromatographic (EC/GC) methods were therefore devised for the analysis of sulfamethazine residues in animal feed, human urine, and wastewater at levels as low as 100, 10, and 10 ppb, respectively. Sample extracts were cleaned up by using liquid/liquid partitioning, and the extracts were subjected to two derivatizations followed by cleanup on a silica gel column. The derivatizations of sulfamethazine consisted of methylation followed by trifluoroacetylation of the primary amine function. Ancillary data concerning stability of the compound in animal feed, water, and as a dry residue on glass, extraction efficiencies, partition values with various solvents, and the analysis of residues in feed by high pressure liquid chromatography (HPLC) at levels as low as 1.0 ppm are presented.

  2. Determination of human albumin in serum and urine samples by constant-energy synchronous fluorescence method.

    Science.gov (United States)

    Madrakian, Tayyebeh; Bagheri, Habibollah; Afkhami, Abbas

    2015-08-01

    A sensitive spectrofluorimetric method using constant-energy synchronous fluorescence technique is proposed for the determination of human albumin without separation. In this method, no reagent was used for enhancement of the fluorescence signal of albumin in the solution. Effects of some parameters, such as energy difference between excitation and emission monochromators (ΔE), emission and excitation slit widths and scan rate of wavelength were studied and the optimum conditions were established. For this purpose factorial design and response surface method were employed for optimization of the effective parameters on the fluorescence signal. The results showed that the scan rate of the wavelength has no significant effect on the analytical signal. The calibration curve was linear in the range 0.1-220.0 µg mL(-1) of albumin with a detection limit of 7.0 × 10(-3)  µg mL(-1). The relative standard deviations (RSD) for six replicate measurements of albumin were calculated as 2.2%, 1.7% and 1.3% for 0.5, 10.0 and 100.0 µg mL(-1) albumin, respectively. Furthermore the proposed method has been employed for the determination of albumin in human serum and urine samples.

  3. Calcium isolation from large-volume human urine samples for 41Ca analysis by accelerator mass spectrometry.

    Science.gov (United States)

    Miller, James J; Hui, Susanta K; Jackson, George S; Clark, Sara P; Einstein, Jane; Weaver, Connie M; Bhattacharyya, Maryka H

    2013-08-01

    Calcium oxalate precipitation is the first step in preparation of biological samples for (41)Ca analysis by accelerator mass spectrometry. A simplified protocol for large-volume human urine samples was characterized, with statistically significant increases in ion current and decreases in interference. This large-volume assay minimizes cost and effort and maximizes time after (41)Ca administration during which human samples, collected over a lifetime, provide (41)Ca:Ca ratios that are significantly above background.

  4. Calcium Isolation from Large-Volume Human Urine Samples for 41Ca Analysis by Accelerator Mass Spectrometry

    Science.gov (United States)

    Miller, James J; Hui, Susanta K; Jackson, George S; Clark, Sara P; Einstein, Jane; Weaver, Connie M; Bhattacharyya, Maryka H

    2013-01-01

    Calcium oxalate precipitation is the first step in preparation of biological samples for 41Ca analysis by accelerator mass spectrometry. A simplified protocol for large-volume human urine samples was characterized, with statistically significant increases in ion current and decreases in interference. This large-volume assay minimizes cost and effort and maximizes time after 41Ca administration during which human samples, collected over a lifetime, provide 41Ca:Ca ratios that are significantly above background. PMID:23672965

  5. Ebola Virus RNA Stability in Human Blood and Urine in West Africa's Environmental Conditions.

    Science.gov (United States)

    Janvier, Frédéric; Delaune, Deborah; Poyot, Thomas; Valade, Eric; Mérens, Audrey; Rollin, Pierre E; Foissaud, Vincent

    2016-02-01

    We evaluated RNA stability of Ebola virus in EDTA blood and urine samples collected from infected patients and stored in West Africa's environmental conditions. In blood, RNA was stable for at least 18 days when initial cycle threshold values were <30, but in urine, RNA degradation occurred more quickly.

  6. Advances in the Diagnosis of Human Opisthorchiasis: Development of Opisthorchis viverrini Antigen Detection in Urine.

    Directory of Open Access Journals (Sweden)

    Chanika Worasith

    Full Text Available Many strategies to control opisthorchiasis have been employed in Thailand, but not in the other neighbouring countries. Specific control methods include mass drug administration (MDA and health education to reduce raw fish consumption. These control efforts have greatly shifted the epidemiology of Opisthorchis viverrini (OV infection over the last decade from presenting as densely concentrated "heavy" infections in single villages to widespread "light" OV infections distributed over wide geographical areas. Currently, the "gold standard" detection method for OV infection is formalin ethyl-acetate concentration technique (FECT, which has limited diagnostic sensitivity and diagnostic specificity for light OV infections, with OV eggs often confused with eggs of minute intestinal flukes (MIFs in feces. In this study, we developed and evaluated the diagnostic performance of a monoclonal antibody-based enzyme-linked immunosorbent assay for the measurement of OV excretory-secretory (ES antigens in urine (urine OV-ES assay for the diagnosis of opisthorchiasis compared to the gold standard detection FECT method.We tested several methods for pre-treating urine samples prior to testing the diagnostic performance of the urine OV-ES assay. Using trichloroacetic acid (TCA pre-treated urine, we compared detection and quantification of OV infection using the urine OV-ES assay versus FECT in OV-endemic areas in Northeastern Thailand. Receiver operating characteristic (ROC curves were used to determine the diagnostic sensitivity and specificity of the urine OV-ES assay using TCA pre-treated urine, and to establish diagnostic positivity thresholds. The Positive Predictive Value as well as the likelihood of obtaining a positive test result (LR+ or a negative test result (LR- were calculated for the established diagnostic positivity threshold. Diagnostic risks (Odds Ratios were estimated using logistic regression.When urine samples were pre-treated with TCA prior to

  7. [Direct proteomic profiling of human urine and blood serum in an experiment with 5-day dry immersion].

    Science.gov (United States)

    2012-01-01

    Changes in proteome of urine and blood serum obtained from 14 healthy humans (age 21-29 yrs) medically certified for an experiment with dry immersion were analyzed. Urine and serum samples were pre-fractionated and enriched with magnetic particles MB-WCX and MB-HIC, respectively, on robot ClinProt (Bruker Daltonics) for direct mass-spectrometry profiling by MALDI-TOF. As a result, 143 protein peaks on the average were identified in urine samples. It was shown that a high variation coefficient in 23.7% of protein peaks, i.e. double technical, points to the most plastic fraction of the urine proteome. In blood serum, 175 peaks were identified in a sample on the average. Comparison of baseline and immersion mass-spectra of the blood proteome revealed significant differences. Increased peak areas of several protein fragments--C3 and C4 fragments of complement system, high-molecular kininogen and fibrinogen--can be ascribed to human body adaptation to the experimental conditions.

  8. Determination of gouty arthritis' biomarkers in human urine using reversed-phase high-performance liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    Lei-Wen Xiang; Jing Li; Jin-Ming Lin; Hai-Fang Li

    2014-01-01

    Creatinine, uric acid, hypoxanthine and xanthine are important diagnostic biomarkers in human urine for gouty arthritis or renal disease diacrisis. A simple method for simultaneous determination of these biomarkers in urine based on reversed-phase high-performance liquid chromatography (RP-HPLC) with ultraviolet (UV) detector was proposed. After pretreatment by dilution, centrifugation and filtration, the biomarkers in urine samples were separated by ODS-BP column by elution with methanol/50 mM NaH2PO4 buffer solution at pH 5.26 (5:95). Good linearity between peak areas and concentrations of standards was obtained for the biomarkers with correlation coefficients in the range of 0.9957-0.9993. The proposed analytical method has satisfactory repeatability (the recovery of data in a range of creatinine, uric acid, hypoxanthine and xanthine was 93.49-97.90%, 95.38-96.45%, 112.46-115.78%and 90.82-97.13%with standard deviation of o5%, respectively) and the limits of detection (LODs, S/N Z 3) for creatinine, uric acid, hypoxanthine, and xanthine were 0.010, 0.025, 0.050 and 0.025 mg/L, respectively. The established method was proved to be simple, accurate, sensitive and reliable for the quantitation of gouty arthritis' biomarkers in human urine samples. The ratio of creatinine to uric acid was found to be a possible factor for assessment of gouty arthritis.

  9. Portable kit for identification and detection of drugs in human urine using surface-enhanced Raman spectroscopy.

    Science.gov (United States)

    Han, Zhenzhen; Liu, Honglin; Meng, Juan; Yang, Liangbao; Liu, Jing; Liu, Jinhuai

    2015-09-15

    A portable kit was demonstrated for rapid and reliable surface-enhanced Raman scattering (SERS) detection of drugs in human urine. This kit contains two sealed reagent tubes, a packet of standardized SERS substrates, and a mini Raman device. A 3 min pretreatment for separating amphetamines from human urine was developed with an extraction rate of >80% examined by ultraperformance liquid chromatography (UPLC). Simultaneously, highly reproducible two-dimensional (2D) gold nanorod (GNR) arrays were assembled by the use of methoxymercaptopoly(ethylene glycol) (mPEG-SH) capping. Thirty batches of GNR arrays produced the 1001 cm(-1) intensity of methamphetamine (MA) molecules with a relative standard deviation (RSD) of 7.9%, and a 21 × 21 μm(2) area mapping on a 2D GNR array produced a statistical RSD of amphetamines in human urine was at least 0.1 ppm. Moreover, the portable kit was successfully used for detecting MA, 3,4-methylenedioxymethamphetamine (MDMA), and methcathinone (MC) in 30 volunteers' urine samples with various clinical natures, and the dual-analyte detection of MA and MDMA implied a good capability of multiplex analysis. UPLC examination and the SERS recovery test clearly indicated that our pretreatment procedure was sufficient to lower the high background signals caused by complex components in urine and demonstrated the practicability and the resistance to false positives, which is a vital problem for law enforcement applications. The excellent performance of our portable kit promises a great prospective toward a rapid, reliable, and on-spot analyzer, especially for public safety and healthcare.

  10. Dystrophin-deficient cardiomyocytes derived from human urine: New biologic reagents for drug discovery

    Directory of Open Access Journals (Sweden)

    Xuan Guan

    2014-03-01

    Full Text Available The ability to extract somatic cells from a patient and reprogram them to pluripotency opens up new possibilities for personalized medicine. Induced pluripotent stem cells (iPSCs have been employed to generate beating cardiomyocytes from a patient's skin or blood cells. Here, iPSC methods were used to generate cardiomyocytes starting from the urine of a patient with Duchenne muscular dystrophy (DMD. Urine was chosen as a starting material because it contains adult stem cells called urine-derived stem cells (USCs. USCs express the canonical reprogramming factors c-myc and klf4, and possess high telomerase activity. Pluripotency of urine-derived iPSC clones was confirmed by immunocytochemistry, RT-PCR and teratoma formation. Urine-derived iPSC clones generated from healthy volunteers and a DMD patient were differentiated into beating cardiomyocytes using a series of small molecules in monolayer culture. Results indicate that cardiomyocytes retain the DMD patient's dystrophin mutation. Physiological assays suggest that dystrophin-deficient cardiomyocytes possess phenotypic differences from normal cardiomyocytes. These results demonstrate the feasibility of generating cardiomyocytes from a urine sample and that urine-derived cardiomyocytes retain characteristic features that might be further exploited for mechanistic studies and drug discovery.

  11. Estimation of D-Arabinose by Gas Chromatography/Mass Spectrometry as Surrogate for Mycobacterial Lipoarabinomannan in Human Urine.

    Directory of Open Access Journals (Sweden)

    Prithwiraj De

    Full Text Available Globally, tuberculosis is slowly declining each year and it is estimated that 37 million lives were saved between 2000 and 2013 through effective diagnosis and treatment. Currently, diagnosis relies on demonstration of the bacteria, Mycobacterium tuberculosis (Mtb, in clinical specimens by serial sputum microscopy, culture and molecular testing. Commercial immunoassay lateral flow kits developed to detect Mtb lipoglycan lipoarabinomannan (LAM in urine as a marker of active TB exhibit poor sensitivity, especially in immunocompetent individuals, perhaps due to low abundance of the analyte. Our present study was designed to develop methods to validate the presence of LAM in a quantitative fashion in human urine samples obtained from culture-confirmed TB patients. Herein we describe, a consolidated approach for isolating LAM from the urine and quantifying D-arabinose as a proxy for LAM, using Gas Chromatography/Mass Spectrometry. 298 urine samples obtained from a repository were rigorously analyzed and shown to contain varying amounts of LAM-equivalent ranging between ~10-40 ng/mL. To further substantiate that D-arabinose detected in the samples originated from LAM, tuberculostearic acid, the unique 10-methyloctadecanoic acid present at the phosphatidylinositol end of LAM was also analyzed in a set of samples and found to be present confirming that the D-arabinose was indeed derived from LAM. Among the 144 samples from culture-negative TB suspects, 30 showed presence of D-arabinose suggesting another source of the analyte, such as disseminated TB or from non-tuberculosis mycobacterium. Our work validates that LAM is present in the urine samples of culture-positive patients in small but readily detectable amounts. The study further substantiates LAM in urine as a powerful biomarker for active tuberculosis.

  12. Colony stimulating factor (CSF) from human leukemic urine: affinity chromatography and isoelectric focusing.

    Science.gov (United States)

    Kellar, K L; Vogler, W R; Kinkade, J M

    1975-12-01

    Biological activities associated with colony-stimulating factor (CSF) from human leukemic urine were found to be selectively retained on an affinity adsorbent of Con A-Sepharose. Elution of activity was achieved using a linear gradient of eith alpha-methyl-D-mannopyranoside (alphaMM) or alpha-methyl-D-glucopyranoside (alphaMG), and resulted in significant increases in specific biological activity. Rechromatography of appropriate fractions indicated that retention of CSF activities was not artifactual. Pretreatment of the affinity matrix with alphaMM completely inhibited binding of CSF. Affinity chromatography of CSF on a Blue Dextran-Sepharose adsorbent was found to be an effective method for removing albumin, a major protein contaminant in urinary preparations. Treatment of CSF with neuraminidase had no effect on its in vitro activity; however, such treatment resulted in an increase in the isoelectric point of CSF from pH 3.5 to pH 4.7, as determined using both sucrose and polyacrylamide gel stabilized pH gradients. Relatively broad regions of biological activity were observed following isoelectric focusing of both neuraminidase-treated and untreated CSF, suggesting that activity was associated with a heterogeneous/polydisperse population of molecular species.

  13. An environmentally friendly reflectometric method for ranitidine determination in pharmaceuticals and human urine

    Science.gov (United States)

    Lima, Liliane Spazzapam; Weinert, Patrícia Los; Lemos, Sahra Cavalcante; Sequinel, Rodrigo; Pezza, Helena Redigolo; Pezza, Leonardo

    2009-01-01

    This paper describes an environmentally friendly method for quantitative determination of ranitidine using diffuse reflectance spectroscopy. This method is based on the reflectance measurements of the colored product produced from the spot test reaction between ranitidine and p-dimethylaminocinnamaldehyde ( p-DAC), in acid medium, using filter paper as solid support. Experimental design methodologies were used to optimize the optimal conditions. All reflectance measurements were carried out at 590 nm and the linear range was from 1.42 × 10 -3 to 3.42 × 10 -2 mol L -1, with a correlation coefficient of 0.997. The limit of detection was estimated to be 1.09 × 10 -3 mol L -1 (R.S.D. = 1.9%). The proposed method was successfully applied to the determination of ranitidine in commercial brands of pharmaceuticals and no interferences were observed from the common excipients in formulations. The results obtained by the proposed method were favorably compared with those obtained by an official procedure at 95% confidence level. Additionally, the method was also applied to the determination of ranitidine in human urine showing excellent recoveries (99.6-100.3%).

  14. Molecular Detection of Campylobacter spp. in California Gull (Larus californicus) Excreta

    Science.gov (United States)

    We examined the prevalence, quantity, and diversity of Campylobacter species in the excreta of 159 California gull samples using PCR and qPCR based detection assays. While Campylobacter prevalence and abundance was relatively high in the gull excreta examined, molecular data ind...

  15. Molecular Detection of Campylobacter spp. in California Gull (Larus californicus) Excreta

    Science.gov (United States)

    We examined the prevalence, quantity, and diversity of Campylobacter species in the excreta of 159 California gull samples using PCR and qPCR based detection assays. While Campylobacter prevalence and abundance was relatively high in the gull excreta examined, molecular data ind...

  16. Methamphetamine and Amphetamine Isomer Concentrations in Human Urine Following Controlled Vicks VapoInhaler Administration

    OpenAIRE

    Smith, Michael L.; Nichols, Daniel C.; Underwood, Paula; Fuller, Zachary; Moser, Matthew A.; Flegel, Ron; Gorelick, David A.; Newmeyer, Matthew N.; Concheiro, Marta; HUESTIS, MARILYN A.

    2014-01-01

    Legitimate use of legal intranasal decongestants containing l-methamphetamine may complicate interpretation of urine drug tests positive for amphetamines. Our study hypotheses were that commonly used immunoassays would produce no false-positive results and a recently developed enantiomer-specific gas chromatography–mass spectrometry (GC–MS) procedure would find no d-amphetamine or d-methamphetamine in urine following controlled Vicks VapoInhaler administration at manufacturer's recommended do...

  17. An optical spot test for the detection of dopamine in human urine using stabilized in air lipid films.

    Science.gov (United States)

    Nikolelis, Dimitrios P; Drivelos, Dimitrios A; Simantiraki, Maria G; Koinis, Spyros

    2004-04-15

    The present technique describes a simple, sensitive spot test for the rapid one-shot detection of dopamine in human urine using lipid films with incorporated resorcin[4]arene receptor that are synthesized by a chemical reaction with a methacrylate polymer on a glass fiber filter. The lipid films without the receptor provided fluorescence under a UV lamp. The use of the receptor in these films quenched this fluorescence, and the color became similar to that of the filters without the lipid films. A drop of dopamine or urine containing this stimulant provided a "switching on" of the fluorescence, which allows the rapid detection of this stimulant in human urine at 10(-8) M concentrations. The novelty of the present work is that it opens new routes in the field of biosensing, i.e., development of sensitive, rapid, and simple methods for detecting species based on the fluorescence of the lipid membranes on a polymer film, and provides a spot test technique for the rapid detection of dopamine. The effect of potent interferences including a wide range of compounds usually found in human urine (i.e., ascorbic aid, glucose, leucine, glycine, tartrate, citrate, bicarbonate, and caffeine) was examined using an aqueous buffered solution that contained the potent interference and dopamine at two lower concentration levels (i.e., 3 x 10(-8)-10(-8) M). The effect of proteins and lipids was also investigated at these two lower dopamine concentration levels in aqueous buffered solution. The results showed no interferences from all these constituents at concentrations usually found in human urine samples; for example, albumin up to 3.22 g/L concentration levels did not provide any interference (i.e., no fluorescence). A drop of urine containing this stimulant provided similar results, i.e., a "switching on" of the fluorescence that allows a technique for the rapid detection of this stimulant in human urine at 10(-8) M concentrations. The technique is not based on a calibration

  18. Simultaneous Determination of Tramadol and Its Metabolite in Human Urine by the Gas Chromatography-Mass Spectrometry Method.

    Science.gov (United States)

    Yilmaz, Bilal; Erdem, Ali Fuat

    2015-08-01

    A sensitive and efficient method was developed for determination of tramadol and its metabolite (O-desmethyltramadol) in human urine by gas chromatography-mass spectrometry. Tramadol, O-desmethyltramadol and medazepam (internal standard) were extracted from human urine with a mixture of ethylacetate and diethylether mixture (1 : 1, v/v) at basic pH with liquid-liquid extraction. The calibration curves were linear (r = 0.99) over tramadol and O-desmethyltramadol concentrations ranging from 10 to 200 ng/mL and 7.5 to 300 ng/mL, respectively. The method had an accuracy of >95% and intra- and interday precision (relative standard deviation %) of ≤4.93 and ≤4.62% for tramadol and O-desmethyltramadol, respectively. The extraction recoveries were found to be 94.1 ± 2.91 and 96.3 ± 3.46% for tramadol and O-desmethyltramadol, respectively. The limit of quantification using 0.5 mL human urine was 10 ng/mL for tramadol and 7.5 ng/mL for O-desmethyltramadol. After oral administration of 100 mg of tramadol hydrochloride to a patient, the urinary excretion was monitored during 24 h. About 15% of the dose was excreted as unchanged tramadol. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  19. Sensitive spectrophotometric determination of metoclopramide hydrochloride in dosage forms and spiked human urine using vanillin

    Directory of Open Access Journals (Sweden)

    O. Zenita Devi

    2016-09-01

    Full Text Available A new spectrophotometric method which is simple, sensitive, selective and rapid is described for the determination of metoclopramide hydrochloride (MCP in bulk drug and in dosage forms using vanillin as the chromogenic agent. The method is based on the condensation reaction between primary aromatic amine group present in MCP with aromatic aldehyde, vanillin to produce an intense yellow colored product. The resulting Schiff’s base shows an absorption maximum at 410 nm and the reaction product is stable for more than one day. The reaction was carried out in acetic acid and perchloric acid medium. Beer’s law was obeyed in the concentration range 1.5–15.0 μg ml−1 MCP with a molar absorptivity of 1.89 × 104 l mol−1 cm−1. The limit of detection (LOD and limit of quantification (LOQ were found to be 0.51 and 1.55 μg ml−1, respectively. The method was statistically evaluated by calculating percent relative error (% RE for accuracy and percent relative standard deviation (% RSD for precision, and was applied successfully to the determination of MCP in tablets, in injection and also in spiked human urine. No interference was observed from common additives found in pharmaceutical preparations. The results obtained by the proposed method were validated statistically by comparing the results with those of the reference method by applying the Student’s t-test and F-test. The accuracy and reliability of the method were further ascertained by performing recovery tests via standard-addition technique.

  20. Qualitative profiling and quantification of neonicotinoid metabolites in human urine by liquid chromatography coupled with mass spectrometry.

    Directory of Open Access Journals (Sweden)

    Kumiko Taira

    Full Text Available Neonicotinoid pesticides have been widely applied for the production of fruits and vegetables, and occasionally detected in conventionally grown produce. Thus oral exposure to neonicotinoid pesticides may exist in the general population; however, neonicotinoid metabolites in human body fluids have not been investigated comprehensively. The purpose of this study is the qualitative profiling and quantitative analysis of neonicotinoid metabolites in the human spot urine by liquid chromatography coupled with mass spectrometry (LC/MS. Human urine samples were collected from three patients suspected of subacute exposure to neonicotinoid pesticides. A qualitative profiling of urinary metabolites was performed using liquid chromatography/time-of-flight mass spectrometry (LC/TOFMS with a database of nominal molecular weights of 57 known metabolites of three neonicotinoid pesticides (acetamiprid, Imidacloprid, and clothianidin, as well as the parent compounds. Then a quantitative analysis of selected urinary metabolites was performed using liquid chromatography/tandem mass spectrometry (LC/MS/MS with a standard pesticide and metabolite, which were detected by the qualitative profiling. The result of qualitative profiling showed that seven metabolites, i.e. an acetamiprid metabolite, N-desmethyl-acetamiprid; three Imidacloprid metabolites, 5-hydroxy-Imidacloprid, 4,5-dihydroxy-imidacloprid, 4,5-dehydro-Imidacloprid; a common metabolite of acetamiprid and Imidacloprid, N-(6-chloronicotinoyl-glycine; and two clothianidin metabolites, N-desmethyl-clothianidin, N-(2-(methylsulfanylthiazole-5-carboxyl-glycine, as well as acetamiprid, were detected in the urine of three cases. The result of the quantitative analysis showed N-desmethyl-acetamiprid was determined in the urine of one case, which had been collected on the first visit, at a concentration of 3.2 ng/mL. This is the first report on the qualitative and quantitative detection of N-desmethyl-acetamiprid in

  1. Development and application of a robust speciation method for determination of six arsenic compounds present in human urine.

    Science.gov (United States)

    Milstein, Lisa S; Essader, Amal; Pellizzari, Edo D; Fernando, Reshan A; Raymer, James H; Levine, Keith E; Akinbo, Olujide

    2003-01-01

    Six arsenic species [arsenate, arsenite, arsenocholine, arsenobetaine, monomethyl arsonic acid, and dimethyl arsinic acid] present in human urine were determined using ion-exchange chromatography combined with inductively coupled plasma mass spectrometry (IC-ICP-MS). Baseline separation was achieved for all six species as well as for the internal standard (potassium hexahydroxy antimonate V) in a single chromatographic run of less than 30 min, using an ammonium carbonate buffer gradient (between 10 and 50 mM) at ambient temperature, in conjunction with cation- and anion-exchange columns in series. The performance of the method was evaluated with respect to linearity, precision, accuracy, and detection limits. This method was applied to determine the concentration of these six arsenic species in human urine samples (n = 251) collected from a population-based exposure assessment survey. Method precision was demonstrated by the analysis of duplicate samples that were prepared over a 2-year analysis period. Total arsenic was also determined for the urine samples using flow injection analysis coupled to ICP-MS. The summed concentration of the arsenic species was compared with the measured arsenic total to demonstrate mass balance. PMID:12611657

  2. Determination of sulpiride in human urine using excitation-emission matrix fluorescence coupled with second-order calibration.

    Science.gov (United States)

    Nie, Jin-Fang; Wu, Hai-Long; Xia, A-Lin; Zhu, Shao-Hua; Bian, Ying-Chao; Li, Shu-Fang; Yu, Ru-Qin

    2007-12-01

    This paper proposes a new and effective approach for the quantitative analysis of sulpiride, a significant antipsychotic drug, in human urine samples by the incorporation of excitation-emission matrix (EEM) fluorescence and second-order calibration methodologies based on the alternating fitting residue (AFR) and self-weighted alternating trilinear decomposition (SWATLD) algorithms. With the application of a second-order advantage, the proposed strategy could be utilized for a direct concentration determination of sulpiride with a simple pretreatment step, even in the presence of serious natural fluorescent interferences. The average recoveries of sulpiride in complex urine samples by using AFR and SWATLD with an estimated component number of three were 101.2 +/- 2.1 and 94.4 +/- 0.7%, respectively. Moreover, the accuracy of the two algorithms was also evaluated through elliptical joint confidence region (EJCR) tests as well as the figures of merit, such as sensitivity (SEN), selectivity (SEL) and limit of detection (LOD). The experimental results demonstrated that both algorithms, as promising quantitative alternatives, have been satisfactorily applied to the determination of sulpiride in human urine, but the performance of AFR was slightly better than that of SWATLD.

  3. Magnetic solid-phase extraction for determination of sulpiride in human urine and blood using high-performance liquid chromatography.

    Science.gov (United States)

    Zhao, Jiao; Liao, Wenlong; Yang, Yaling

    2015-12-01

    A novel and efficient sample preconcentration technique based on the Fe3O4 magnetic nanoparticles (Fe3O4 MNPs) coated with silica (SiO2) has been developed for extraction and determination of sulpiride. The functionalized MNPs showed excellent dispersibility in aqueous solution and were applied to magnetic solid-phase extraction of sulpiride from human urine and blood prior to high-performance liquid chromatography analysis. The separation, preconcentration and desorption procedure was completed in 10 min. Optimal experimental conditions, including sample pH, the amount of the MNPs, eluent type and volume, and the ultrasonication time were studied and established. The method showed good linearity for the determination of sulpiride in the concentration range of 10-1000 ng/mL in urine and blood. The recovery of the method was in the range between 91.2 and 97.5%, and the limit of detection was 2 ng/mL for sulpiride in human blood and urine. The results indicated that the present procedure is a suitable pretreatment method for biological samples.

  4. Removal of ammonium from human urine through ion exchange with clinoptilolite and its recovery for further reuse.

    Science.gov (United States)

    Beler-Baykal, B; Bayram, S; Akkaymak, E; Cinar, S

    2004-01-01

    Ammonium, from separately collected human urine, had been removed through transfer onto the ammonium selective natural zeolite, clinoptilolite, through ion exchange. In the subsequent treatment steps of washing with tap water, ammonium removed from urine was eluted from the surface of the clinoptilolite to be recovered for further reuse. Different quantities of clinoptilolite were used for a survey of the capacity of the zeolite for the process and to identify removal efficiencies based on initial ammonium loads. The highest surface concentration attained under experimental conditions employed was 15.44 mg ammonium per gram of clinoptilolite for an initial concentration of 110 mg ammonia per litre, and the highest removal was 98%, obtained for a loading of 1 mg ammonium per gram clinoptilolite. In the subsequent elution process, better removals were observed as pH was increased and the highest removal was attained at pH 13. The recovery was calculated as 9.73 mg ammonium per gram of clinoptilolite, corresponding to an efficiency of 63% only through washing with tap water. The results have given positive indications for the possibility of using ion exchange with clinoptilolite for the removal of ammonium from human urine and an incentive for improving methods of elution for its recovery for further reuse.

  5. The determination of organophosphonate nerve agent metabolites in human urine by hydrophilic interaction liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Mawhinney, Douglas B; Hamelin, Elizabeth I; Fraser, Rheaclare; Silva, Sathya S; Pavlopoulos, Antonis J; Kobelski, Robert J

    2007-06-01

    A sensitive, robust isotope dilution LC/MS/MS method is presented for the quantitative analysis of human urine for the alkyl methylphosphonic acid metabolites of five organophosphorus nerve agents (VX, rVX or VR, GB or Sarin, GD or Soman, and GF or Cyclosarin). The selective sample preparation method employs non-bonded silica solid-phase extraction and is partially automated. While working with a mobile phase composition that enhances the electrospray ionization process, the hydrophilic interaction chromatography method results in a 5-min injection-to-injection cycle time, excellent peak shapes and adequate retention (k'=3.1). These factors lead to limits of detection for these metabolites as low as 30 pg/mL in a 1-mL sample of human urine. The quality control data (15 and 75 ng/mL) demonstrate accurate (-0.5 to +3.4%) and precise (coefficients of variation of 2.1-3.6%) quantitative results over the clinically relevant urine concentration range of 1-200 ng/mL for a validation set of 20 standard and quality control sets prepared by five analysts over 54 days. The selectivity of the method is demonstrated for a 100-individual reference range study, as well as the analysis of relevant biological samples. The combined sample preparation and analysis portions of this method have a throughput of 288 samples per day.

  6. Simultaneous determination of nabumetone and its principal metabolite in medicines and human urine by time-resolved fluorescence.

    Science.gov (United States)

    Murillo Pulgarín, José Antonio; Alañón Molina, Aurelia; Martínez Ferreras, Fernando

    2012-11-07

    A simple fluorescent methodology for the simultaneous determination of nabumetone and its main metabolite, 6-methoxy-2-naphthylacetic acid (6-MNA), in pharmaceutical preparations and human urine is proposed. Due to the strong overlapping between the fluorescence spectra of both analytes, the use of fluorescence decay curves to resolve their mixture is proposed, since these curves are more selective. Values of dependent instrumental variables affecting the signal-to-noise ratio were fixed using a simplex optimization procedure. A factorial design with three levels per factor coupled to a central composite design was selected to obtain a calibration matrix of thirteen standards plus one blank sample that was processed using a partial least-squares (PLS) analysis. In order to assess the goodness of the proposed method, a prediction set of ten synthetic samples was analyzed, obtaining recovery percentages between 97 and 105%. Limits of detection, calculated by means of a new criterion, were 0.96 μg L(-1) and 0.88 μg L(-1) for nabumetone and 6-MNA, respectively. The method was also tested in the pharmaceutical preparation Relif, which contains nabumetone, obtaining recovery percentages close to 100%. Finally, the simultaneous determination of both analytes in human urine samples was successfully carried out by the PLS-analysis of a matrix of fifteen standards plus four analyte blanks and the use of the standard addition technique. Although urine shows native fluorescence, no extraction method or prior separation of the analytes was needed.

  7. Detection of renal tissue and urinary tract proteins in the human urine after space flight.

    Directory of Open Access Journals (Sweden)

    Lyudmila Kh Pastushkova

    Full Text Available The urine protein composition samples of ten Russian cosmonauts (male, aged of 35 up to 51 performed long flight missions and varied from 169 up to 199 days on the International Space Station (ISS were analyzed. As a control group, urine samples of six back-up cosmonauts were analyzed. We used proteomic techniques to obtain data and contemporary bioinformatics approaches to perform the analysis. From the total number of identified proteins (238 in our data set, 129 were associated with a known tissue origin. Preflight samples contained 92 tissue-specific proteins, samples obtained on Day 1 after landing had 90 such proteins, while Day 7 samples offered 95 tissue-specific proteins. Analysis showed that consistently present proteins in urine (under physiological conditions and after space flight are cubilin, epidermal growth factor, kallikrein-1, kininogen-1, megalin, osteopontin, vitamin K-dependent protein Z, uromodulin. Variably present proteins consists of: Na(+/K(+ ATPase subunit gamma, β-defensin-1, dipeptidyl peptidase 4, maltasa-glucoamilasa, cadherin-like protein, neutral endopeptidase and vascular cell adhesion protein 1. And only three renal proteins were related to the space flight factors. They were not found in the pre-flight samples and in the back-up cosmonaut urine, but were found in the urine samples after space flight: AFAM (afamin, AMPE (aminopeptidase A and AQP2 (aquaporin-2. This data related with physiological readaptation of water-salt balance. The proteomic analysis of urine samples in different phases of space missions with bioinformation approach to protein identification provides new data relative to biomechemical mechanism of kidney functioning after space flight.

  8. The effect of water, ascorbic acid, and cranberry derived supplementation on human urine and uropathogen adhesion to silicone rubber

    NARCIS (Netherlands)

    Habash, MB; Van der Mei, HC; Busscher, HJ; Reid, G

    1999-01-01

    In this study, urine was collected from groups of volunteers following the consumption of water, ascorbic acid, or cranberry supplements. Only ascorbic acid intake consistently produced acidic urine. Photospectroscopy data indicated that increased water consumption produced urine with lower protein

  9. Designer phenethylamines routinely found in human urine: 2-ethylamino-1-phenylbutane and 2-amino-1-phenylbutane.

    Science.gov (United States)

    Uralets, Victor; App, Mike; Rana, Sumandeep; Morgan, Stewart; Ross, Wayne

    2014-03-01

    2-Ethylamino-1-phenylbutane (EAPB) and 2-amino-1-phenylbutane (APB) were identified by gas chromatography-mass spectrometry in multiple urine samples submitted for stimulant drug testing and screened positive for amphetamines by enzyme immunoassay. Forty-two samples from all over the USA were found, containing both analytes during a 3-month period May-July 2013. A sports dietary supplement 'CRAZE' has been determined to be one of the sources of EAPB supply. EAPB along with its suggested metabolite APB were detected in a urine sample, obtained from a person known to use 'CRAZE'.

  10. Rapid simultaneous determination of ephedrines, amphetamines, cocaine, cocaine metabolites, and opiates in human urine by GC-MS.

    Science.gov (United States)

    Saito, Takeshi; Mase, Hiroyasu; Takeichi, Sanae; Inokuchi, Sadaki

    2007-01-04

    This paper presents a simple and sensitive chromatographic procedure for the simultaneous determination and quantification of ephedrines, amphetamines, cocaine, cocaine metabolites, and opiates in human urine by gas chromatography-mass spectrometry. This method involves enzyme hydrolysis in the presence of a deuterated internal standard, liquid-liquid extraction, and derivatization with pentafluoropropionic anhydride and pentafluoropropanol. The recovery of each compound averaged at 65.8% or more. The limits of detection determined for each compound by using a 2-mL sample volume ranged from 5 to 50 ng/mL. The calibration curves were linear to 100 ng/mL for all compounds when determined using methamphetamine-d4 and MDMA-d5 as internal standards. This method was successfully applied for the analysis of urine samples suspected to contain intoxicants such as methamphetamine and heroin.

  11. Determination of gouty arthritis' biomarkers in human urine using reversed-phase high-performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Lei-Wen Xiang

    2014-04-01

    Full Text Available Creatinine, uric acid, hypoxanthine and xanthine are important diagnostic biomarkers in human urine for gouty arthritis or renal disease diacrisis. A simple method for simultaneous determination of these biomarkers in urine based on reversed-phase high-performance liquid chromatography (RP-HPLC with ultraviolet (UV detector was proposed. After pretreatment by dilution, centrifugation and filtration, the biomarkers in urine samples were separated by ODS-BP column by elution with methanol/50 mM NaH2PO4 buffer solution at pH 5.26 (5:95. Good linearity between peak areas and concentrations of standards was obtained for the biomarkers with correlation coefficients in the range of 0.9957–0.9993. The proposed analytical method has satisfactory repeatability (the recovery of data in a range of creatinine, uric acid, hypoxanthine and xanthine was 93.49–97.90%, 95.38–96.45%, 112.46–115.78% and 90.82–97.13% with standard deviation of <5%, respectively and the limits of detection (LODs, S/N≥3 for creatinine, uric acid, hypoxanthine, and xanthine were 0.010, 0.025, 0.050 and 0.025 mg/L, respectively. The established method was proved to be simple, accurate, sensitive and reliable for the quantitation of gouty arthritis' biomarkers in human urine samples. The ratio of creatinine to uric acid was found to be a possible factor for assessment of gouty arthritis.

  12. Influence of multiple injections of human chorionic gonadotropin (hCG) on urine and serum endogenous steroids concentrations.

    Science.gov (United States)

    Strahm, Emmanuel; Marques-Vidal, Pedro; Pralong, François; Dvorak, Jiri; Saugy, Martial; Baume, Norbert

    2011-12-10

    Since it is established that human chorionic gonadotropin (hCG) affects testosterone production and release in the human body, the use of this hormone as a performance enhancing drug has been prohibited by the World Anti-Doping Agency. Nowadays, the only validated biomarker of a hCG doping is its direct quantification in urine. However, this specific parameter is subjected to large inter-individual variability and its determination is directly dependent on the reliability of hCG immunoassays used. In order to counteract these weaknesses, new biomarkers need to be evidenced. To address this issue, a pilot clinical study was performed on 10 volunteers submitted to 3 subsequent hCG injections. Blood and urine samples were collected during two weeks in order to follow the physiological effects on related compounds such as the steroid profile or hormones involved in the hypothalamo-pituitary axis. The hCG pharmacokinetic observed in all subjects was, as expected, prone to important inter-individual variations. Using ROC plots, level of testosterone and testosterone on luteinizing hormone ratio in both blood and urine were found to be the most relevant biomarker of a hCG abuse, regardless of inter-individual variations. In conclusion, this study showed the crucial importance of reliable quantification methods to assess low differences in hormonal patterns. In regard to these results and to anti-doping requirements and constraints, blood together with urine matrix should be included in the anti-doping testing program. Together with a longitudinal follow-up approach it could constitute a new strategy to detect a hCG abuse, applicable to further forms of steroid or other forbidden drug manipulation.

  13. Myoglobin urine test

    Science.gov (United States)

    Urine myoglobin; Heart attack - myoglobin urine test; Myositis - myoglobin urine test; Rhabdomyolysis - myoglobin urine test ... The test involves only normal urination, which should cause no discomfort.

  14. HPLC determination of ciprofloxacin, cloxacillin, and ibuprofen drugs in human urine samples.

    Science.gov (United States)

    Espinosa-Mansilla, Anunciación; Muñoz de la Peña, Arsenio; González Gómez, David; Cañada-Cañada, Florentina

    2006-08-01

    This paper reports, for the first time, a liquid chromatographic method for the simultaneous determination of three frequently co-administered active principles, two antibiotics, ciprofloxacin (CIPRO) and cloxacillin (CLOXA) belonging to the fluoroquinolones and beta-lactam families, respectively, and ibuprofen (IBU), a non-steroidal anti-inflammatory drug. The chromatographic separation was performed on a C-18 analytical column, using isocratic elution with methanol-acetonitrile-pH 3 formate buffer (CT = 0.1 M) (15:12:73, v/v/v) for 3 min and, after that, a linear gradient with methanol-acetonitrile (88:12, v/v) for 8 min. Several absorption spectra were obtained for each peak using a DAD detector. Chromatograms at the maximum absorption wavelength for each analyte, 220 nm for both IBU and CLOXA, and 280 nm for CIPRO were selected as the most suitable. The proposed chromatographic method requires about 15 min per sample. The presence of a urine background was tested and no interference was found. The method was satisfactorily applied to the determination of CIPRO, CLOXA, and IBU, in fortified urine, and in urine samples from a patient undergoing treatment with these three active principles, among others. Limits of quantification in urine were 1.00, 1.70, and 2.87 microg/mL for CIPRO, CLOXA, and IBU, respectively.

  15. Quantifying creatinine and urea in human urine through Raman spectroscopy aiming at diagnosis of kidney disease.

    Science.gov (United States)

    Saatkamp, Cassiano Junior; de Almeida, Maurício Liberal; Bispo, Jeyse Aliana Martins; Pinheiro, Antonio Luiz Barbosa; Fernandes, Adriana Barrinha; Silveira, Landulfo

    2016-03-01

    Due to their importance in the regulation of metabolites, the kidneys need continuous monitoring to check for correct functioning, mainly by urea and creatinine urinalysis. This study aimed to develop a model to estimate the concentrations of urea and creatinine in urine by means of Raman spectroscopy (RS) that could be used to diagnose kidney disease. Midstream urine samples were obtained from 54 volunteers with no kidney complaints. Samples were subjected to a standard colorimetric assay of urea and creatinine and submitted to spectroscopic analysis by means of a dispersive Raman spectrometer (830 nm, 350 mW, 30 s). The Raman spectra of urine showed peaks related mainly to urea and creatinine. Partial least squares models were developed using selected Raman bands related to urea and creatinine and the biochemical concentrations in urine measured by the colorimetric method, resulting in r = 0.90 and 0.91 for urea and creatinine, respectively, with root mean square error of cross-validation (RMSEcv) of 312 and 25.2 mg/dL, respectively. RS may become a technique for rapid urinalysis, with concentration errors suitable for population screening aimed at the prevention of renal diseases.

  16. Quantifying creatinine and urea in human urine through Raman spectroscopy aiming at diagnosis of kidney disease

    Science.gov (United States)

    Saatkamp, Cassiano Junior; de Almeida, Maurício Liberal; Bispo, Jeyse Aliana Martins; Pinheiro, Antonio Luiz Barbosa; Fernandes, Adriana Barrinha; Silveira, Landulfo, Jr.

    2016-03-01

    Due to their importance in the regulation of metabolites, the kidneys need continuous monitoring to check for correct functioning, mainly by urea and creatinine urinalysis. This study aimed to develop a model to estimate the concentrations of urea and creatinine in urine by means of Raman spectroscopy (RS) that could be used to diagnose kidney disease. Midstream urine samples were obtained from 54 volunteers with no kidney complaints. Samples were subjected to a standard colorimetric assay of urea and creatinine and submitted to spectroscopic analysis by means of a dispersive Raman spectrometer (830 nm, 350 mW, 30 s). The Raman spectra of urine showed peaks related mainly to urea and creatinine. Partial least squares models were developed using selected Raman bands related to urea and creatinine and the biochemical concentrations in urine measured by the colorimetric method, resulting in r=0.90 and 0.91 for urea and creatinine, respectively, with root mean square error of cross-validation (RMSEcv) of 312 and 25.2 mg/dL, respectively. RS may become a technique for rapid urinalysis, with concentration errors suitable for population screening aimed at the prevention of renal diseases.

  17. The Characterization of Feces and Urine: A Review of the Literature to Inform Advanced Treatment Technology.

    Science.gov (United States)

    Rose, C; Parker, A; Jefferson, B; Cartmell, E

    2015-09-02

    The safe disposal of human excreta is of paramount importance for the health and welfare of populations living in low income countries as well as the prevention of pollution to the surrounding environment. On-site sanitation (OSS) systems are the most numerous means of treating excreta in low income countries, these facilities aim at treating human waste at source and can provide a hygienic and affordable method of waste disposal. However, current OSS systems need improvement and require further research and development. Development of OSS facilities that treat excreta at, or close to, its source require knowledge of the waste stream entering the system. Data regarding the generation rate and the chemical and physical composition of fresh feces and urine was collected from the medical literature as well as the treatability sector. The data were summarized and statistical analysis was used to quantify the major factors that were a significant cause of variability. The impact of this data on biological processes, thermal processes, physical separators, and chemical processes was then assessed. Results showed that the median fecal wet mass production was 128 g/cap/day, with a median dry mass of 29 g/cap/day. Fecal output in healthy individuals was 1.20 defecations per 24 hr period and the main factor affecting fecal mass was the fiber intake of the population. Fecal wet mass values were increased by a factor of 2 in low income countries (high fiber intakes) in comparison to values found in high income countries (low fiber intakes). Feces had a median pH of 6.64 and were composed of 74.6% water. Bacterial biomass is the major component (25-54% of dry solids) of the organic fraction of the feces. Undigested carbohydrate, fiber, protein, and fat comprise the remainder and the amounts depend on diet and diarrhea prevalence in the population. The inorganic component of the feces is primarily undigested dietary elements that also depend on dietary supply. Median urine

  18. Degradation of pharmaceuticals and metabolite in synthetic human urine by UV, UV/H2O2, and UV/PDS.

    Science.gov (United States)

    Zhang, Ruochun; Sun, Peizhe; Boyer, Treavor H; Zhao, Lin; Huang, Ching-Hua

    2015-03-03

    To minimize environmental pharmaceutical micropollutants, treatment of human urine could be an efficient approach due to the high pharmaceutical concentration and toxic potential excreted in urine. This study investigated the degradation kinetics and mechanisms of sulfamethoxazole (SMX), trimethoprim (TMP) and N4-acetyl-sulfamethoxazole (acetyl-SMX) in synthetic fresh and hydrolyzed human urines by low-pressure UV, and UV combined with H2O2 and peroxydisulfate (PDS). The objective was to compare the two advanced oxidation processes (AOPs) and assess the impact of urine matrices. All three compounds reacted quickly in the AOPs, exhibiting rate constants of (6.09-8.53) × 10(9) M(-1)·s(-1) with hydroxyl radical, and (2.35-16.1) × 10(9) M(-1)·s(-1) with sulfate radical. In fresh urine matrix, the pharmaceuticals' indirect photolysis was significantly suppressed by the scavenging effect of urine citrate and urea. In hydrolyzed urine matrix, the indirect photolysis was strongly affected by inorganic urine constituents. Chloride had no apparent impact on UV/H2O2, but significantly raised the hydroxyl radical concentration in UV/PDS. Carbonate species reacted with hydroxyl or sulfate radical to generate carbonate radical, which degraded SMX and TMP, primarily due to the presence of aromatic amino group(s) (k = 2.68 × 10(8) and 3.45 × 10(7) M(-1)·s(-1)) but reacted slowly with acetyl-SMX. Ammonia reacted with hydroxyl or sulfate radical to generate reactive nitrogen species that could react appreciably only with SMX. Kinetic simulation of radical concentrations, along with products analysis, helped elucidate the major reactive species in the pharmaceuticals' degradation. Overall, the AOPs' performance was higher in the hydrolyzed urine than fresh urine matrix with UV/PDS better than UV/H2O2, and varied significantly depending on pharmaceutical's structure.

  19. Analysis of mitragynine and metabolites in human urine for detecting the use of the psychoactive plant kratom.

    Science.gov (United States)

    Le, David; Goggin, Melissa M; Janis, Gregory C

    2012-01-01

    The leaves of the South Asian plant kratom are described as having stimulating effects at low doses, and opiate-like analgesic and euphoric effects at high doses. A long history of use and abuse has led to the classification of kratom as a controlled substance in its native Thailand and other South Asian countries. However, kratom is not controlled in the United States, and the ready availability of kratom has led to its emergence as an herbal drug of abuse. With the growing popularity of kratom, efficient procedures are needed to detect kratom use. In the current study, both ultra-high-performance liquid chromatography and high-performance liquid chromatography-tandem mass spectrometry methods have been developed and validated for monitoring the major alkaloids and metabolites found in urine following kratom use. The primary unique alkaloid mitragynine is quantified in human urine from 1.00-500.00 ng/mL using mitraphylline as an internal standard. In addition, two metabolites (5-desmethylmitragynine and 17-desmethyldihydromitragynine) and the related active, alkaloid 7-hydroxy-mitragynine, are simultaneously qualitatively monitored. The presence of analytes are confirmed by an information-dependent acquisition-enhanced product ion procedure generating full fragmentation data used to positively identify detected analytes. The validated method has been utilized for clinical and forensic analyses of urine for the detection of kratom use.

  20. Methodology for and the determination of the major constituents and metabolites of the Amazonian botanical medicine ayahuasca in human urine.

    Science.gov (United States)

    McIlhenny, Ethan H; Riba, Jordi; Barbanoj, Manel J; Strassman, Rick; Barker, Steven A

    2011-09-01

    Ayahuasca, also known as caapi or yage among various South American groups, holds a highly esteemed and millennia-old position in these cultures' medical and religious pharmacopeia. There is now an increasing interest in the potential for modern medical applications of ayahuasca, as well as concerns regarding its increasing potential for abuse. Toxicological and clinical research to address these issues will require information regarding its metabolism and clearance. Thus, a rapid, sensitive and specific method for characterization and quantitation of the major constituents and of the metabolites of ayahuasca in urine is needed. The present research provides a protocol for conducting such analyses. The characteristics of the method, conducted by sample dilution and using HPLC-electrospray ionization (ESI)-selected reaction monitoring (SRM)-tandem mass spectrometry, are presented. The application of the analytical protocol to urine samples collected from three individuals that were administered ayahuasca has also been demonstrated. The data show that the major metabolite of the hallucinogenic component of ayahuasca, N,N-dimethyltryptamine (DMT), is the corresponding N-oxide, the first time this metabolite has been described in in vivo studies in humans. Further, very little DMT was detected in urine, despite the inhibition of monoamine oxidase afforded by the presence of the harmala alkaloids in ayahuasca. The major harmala alkaloid excreted was tetrahydroharmine. Other excretion products and metabolites were also identified and quantified. The method described would be suitable for use in further toxicological and clinical research on ayahuasca. Copyright © 2010 John Wiley & Sons, Ltd.

  1. Validation of an immunodiagnostic assay for detection of 13 Streptococcus pneumoniae serotype-specific polysaccharides in human urine.

    Science.gov (United States)

    Pride, Michael W; Huijts, Susanne M; Wu, Kangjian; Souza, Victor; Passador, Sherry; Tinder, Chunyan; Song, Esther; Elfassy, Arik; McNeil, Lisa; Menton, Ronald; French, Roger; Callahan, Janice; Webber, Chris; Gruber, William C; Bonten, Marc J M; Jansen, Kathrin U

    2012-08-01

    To improve the clinical diagnosis of pneumococcal infection in bacteremic and nonbacteremic community-acquired pneumonia (CAP), a Luminex technology-based multiplex urinary antigen detection (UAD) diagnostic assay was developed and validated. The UAD assay can simultaneously detect 13 different serotypes of Streptococcus pneumoniae by capturing serotype-specific S. pneumoniae polysaccharides (PnPSs) secreted in human urine. Assay specificity is achieved by capturing the polysaccharides with serotype-specific monoclonal antibodies (MAbs) on spectrally unique microspheres. Positivity for each serotype was based on positivity cutoff values calculated from a standard curve run on each assay plate together with positive- and negative-control urine samples. The assay is highly specific, since significant signals are detected only when each PnPS was paired with its homologous MAb-coated microspheres. Validation experiments demonstrated excellent accuracy and precision. The UAD assay and corresponding positivity cutoff values were clinically validated by assessing 776 urine specimens obtained from patients with X-ray-confirmed CAP. The UAD assay demonstrated 97% sensitivity and 100% specificity using samples obtained from patients with bacteremic, blood culture-positive CAP. Importantly, the UAD assay identified Streptococcus pneumoniae (13 serotypes) in a proportion of individuals with nonbacteremic CAP, a patient population for which the pneumococcal etiology of CAP was previously difficult to assess. Therefore, the UAD assay provides a specific, noninvasive, sensitive, and reproducible tool to support vaccine efficacy as well as epidemiological evaluation of pneumococcal disease, including CAP, in adults.

  2. Analysis of chlorpheniramine in human urine samples using dispersive liquid-liquid microextraction combined with high-performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Mehdi Maham

    2014-09-01

    Full Text Available A simple and environmentally friendly microextraction technique was used for determination of chlorpheniramine (CPM, an antihistamine drug, in human urine samples using dispersive liquid-liquid microextraction (DLLME followed by high performance liquid chromatography with diode array detection (HPLC-DAD. In this extraction technique, an appropriate mixture of acetonitrile (disperser solvent and carbon tetrachloride (extraction solvent was rapidly injected into the urine sample containing the target analyte. Tiny droplets of extractant were formed and dispersed into the sample solution and then sedimented at the bottom of the conical test tube by centrifugation. Under optimal conditions, the calibration curve was linear in the range of 0.055-5.5 µg mL-1, with a detection limit of 16.5 ng mL-1. This proposed method was successfully applied to the analysis of real urine samples. Low consumption of toxic organic solvents, simplicity of operation, low cost and acceptable figures of merit are the main advantages of the proposed technique.

  3. Determination of amphetamines in human urine by liquid chromatography with fluorimetric detection using a solid-phase extraction procedure.

    Science.gov (United States)

    Bugamelli, Francesca; Mandrioli, Roberto; Cavallini, Annalisa; Baccini, Cesare; Conti, Matteo; Raggi, Maria Augusta

    2006-10-01

    A precise and feasible HPLC method has been developed for the analysis of amphetamine (AMPH), methamphetamine (MAMPH) and methylenedioxymethamphetamine (MDMA, ecstasy) in human urine. A chromatographic run on a C8 Genesis (150 mm x 4.6 mm, 5 microm) column maintained at 30 degrees C lasts about 17 min, using a mobile phase composed of ACN (12%) and a pH 2.5 phosphate buffer (88%) containing 0.3% triethylamine. Mirtazapine was used as the internal standard. Good linearity was found in the 100-2000 ng/mL concentration range for AMPH and MAMPH and in the 12-2000 ng/mL concentration range for MDMA. The pretreatment of urine samples was carried out by means of a careful SPE procedure on C2 cartridges. The extraction yields were very satisfactory for all analytes, with average values greater than 97%. The leading conditions allowed the determination of AMPH, MAMPH and MDMA with satisfactory precision and accuracy. The method has been successfully applied to the determination of the analytes in urine of AMPH users.

  4. The use of a gold electrode for the determination of amphetamine derivatives and application to their analysis in human urine

    Directory of Open Access Journals (Sweden)

    Nevešćanin Marina M.

    2013-01-01

    Full Text Available The catalytic abilities of gold electrode were tested for the quantitative determination of amphetamine (A and 3,4-methylenedioxy-N-methylamphetamine (MDMA standards by their oxidation using cyclic voltammetry (CV. The value of the oxidative currents of A and MDMA standards at 0.80 V vs. SCE in 0.05 M NaHCO3 at the scan rate of 50 mV/s is linear function of concentration in range of 110.9-258.9 mM and 38.7-229.2 mM, respectively. Square wave voltammetry (SWV revealed linear increase of current with concentration of MDMA (range 30.9-91.6 mM and thus quantitative determination of amphetamine derivates. SWV analysis is successfully performed in spiked urine samples as well. A and MDMA in the presence of sucrose and as a content in illegally produced tablets were also analyzed. The voltammetric determination of A and MDMA derivatives using CV and SWV at gold electrode is a rapid, selective and simple procedure and its accuracy was confirmed with reference method, high performance liquid chromatography (HPLC. The spiked urine samples analysis offers additional possibility for the rapid detection of A and MDMA in human urine.

  5. Safety assessment of genetically modified rice expressing human serum albumin from urine metabonomics and fecal bacterial profile.

    Science.gov (United States)

    Qi, Xiaozhe; Chen, Siyuan; Sheng, Yao; Guo, Mingzhang; Liu, Yifei; He, Xiaoyun; Huang, Kunlun; Xu, Wentao

    2015-02-01

    The genetically modified (GM) rice expressing human serum albumin (HSA) is used for non-food purposes; however, its food safety assessment should be conducted due to the probability of accidental mixture with conventional food. In this research, Sprague Dawley rats were fed diets containing 50% (wt/wt) GM rice expressing HSA or non-GM rice for 90 days. Urine metabolites were detected by (1)H NMR to examine the changes of the metabolites in the dynamic process of metabolism. Fecal bacterial profiles were detected by denaturing gradient gel electrophoresis to reflect intestinal health. Additionally, short chain fatty acids and fecal enzymes were investigated. The results showed that compared with rats fed the non-GM rice, some significant differences were observed in rats fed with the GM rice; however, these changes were not significantly different from the control diet group. Additionally, the gut microbiota was associated with blood indexes and urine metabolites. In conclusion, the GM rice diet is as safe as the traditional daily diet. Furthermore, urine metabonomics and fecal bacterial profiles provide a non-invasive food safety assessment rat model for genetically modified crops that are used for non-food/feed purposes. Fecal bacterial profiles have the potential for predicting the change of blood indexes in future.

  6. Development of a GC-MS/MS strategy to determine 15 mycotoxins and metabolites in human urine.

    Science.gov (United States)

    Rodríguez-Carrasco, Yelko; Moltó, Juan Carlos; Mañes, Jordi; Berrada, Houda

    2014-10-01

    The widespread mycotoxins contamination of food commodities has made the monitoring of their levels essential. To overcome the disadvantages of the indirect approach by food analysis, detection of mycotoxin as biomarkers in urine provides a useful and specific data for exposure assessment to these food contaminants. In this work, a sensitive, rapid and accurate method based on gas chromatography-tandem mass spectrometry procedure to determine 15 mycotoxins and metabolites in human urine was optimized and validated taking into consideration the guidelines specified in Commission Decision 2002/657/EC and 401/2006/EC. A salting-out assisted acetonitrile-based extraction was used for sample preparation. The extraction recoveries were in a range of 72-109%, with intra-day relative standard deviation and inter-day relative standard deviation lower than 10% and 13%, respectively for all mycotoxins at 50, 100 and 200 µg/L spiking levels. The limits of quantitation ranged from 0.25 to 8 µg/L. Matrix effect was evaluated and matrix-matched calibration was used for quantitation. The proposed procedure was applied to 10 urine samples collected from children. Mycotoxins were quantified in 30% of samples.

  7. Using dispersive liquid-liquid microextraction and liquid chromatography for determination of guaifenesin enantiomers in human urine.

    Science.gov (United States)

    Hatami, Mehdi; Farhadi, Khalil; Abdollahpour, Assem

    2011-11-01

    A simple, rapid, and efficient method, dispersive liquid-liquid microextraction (DLLME) coupled with high-performance liquid chromatography-fluorescence detector, has been developed for the determination of guaifenesin (GUA) enantiomers in human urine samples after an oral dose administration of its syrup formulation. Urine samples were collected during the time intervals 0-2, 2-4, and 4-6 h and concentration and ratio of two enantiomers was determined. The ratio of R-(-) to S-(+) enantiomer concentrations in urine showed an increase with time, with R/S ratios of 0.66 at 2 h and 2.23 at 6 h. For microextraction process, a mixture of extraction solvent (dichloromethane, 100 μL) and dispersive solvent (THF, 1 mL) was rapidly injected into 5.0 mL diluted urine sample for the formation of cloudy solution and extraction of enantiomers into the fine droplets of CH(2)Cl(2). After optimization of HPLC enantioselective conditions, some important parameters, such as the kind and volume of extraction and dispersive solvents, extraction time, temperature, pH, and salt effect were optimized for dispersive liquid-liquid microextraction process. Under the optimum extraction condition, the method yields a linear calibration curve in the concentration range from 10 to 2000 ng/mL for target analytes. LOD was 3.00 ng/mL for both of the enantiomers. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Rapid and sensitive determination of strychnine and brucine in human urine by capillary electrophoresis with field-amplified sample stacking.

    Science.gov (United States)

    Li, Junmei; Jiang, Ye

    2010-02-01

    A simple, rapid, sensitive and low-cost method using capillary electrophoresis (CE) coupled with field-amplified sample stacking (FASS) has been developed and validated for the simultaneous determination of strychnine and brucine residues in human urine. Before sample loading, a water plug (3.5 kPa, 3 s) was injected to contain sample cations and to permit FASS. Electrokinetic injection at a voltage (20 kV, 25 s) was then used to introduce cations. Separation was performed using 20 mM acetate buffer (pH 3.8) with an applied voltage of 20 kV. The calibration curves were linear over a range of 8.00-2.56 infinity 10(2) ng/mL (r = 0.9995) for strychnine and 10.0-3.20 x 10(2) ng/mL (r = 0.9999) for brucine. Extraction recoveries in urine were greater than 79.6 and 82.8% for strychnine and brucine, respectively, with an RSD of less than 4.9%. The detection limits (signal-to-noise ratio 3) for strychnine and brucine were 2.00 and 2.50 ng/mL, respectively. A urine sample from one healthy female volunteer (26 years old, 50 kg) was pretreated and analyzed. Strychnine and brucine levels in urine could be detected 24 h after administration. On these grounds, this method was feasible for application to preliminary screening of trace levels of abused drugs for both doping control and forensic analysis. (c) 2009 John Wiley & Sons, Ltd.

  9. Morphine and Codeine Concentrations in Human Urine following Controlled Poppy Seeds Administration of Known Opiate Content

    Science.gov (United States)

    Smith, Michael L.; Nichols, Daniel C.; Underwood, Paula; Fuller, Zachary; Moser, Matthew A.; LoDico, Charles; Gorelick, David A.; Newmeyer, Matthew N.; Concheiro, Marta; Huestis, Marilyn A.

    2014-01-01

    Opiates are an important component for drug testing due to their high abuse potential. Proper urine opiate interpretation includes ruling out poppy seed ingestion; however, detailed elimination studies after controlled poppy seed administration with known morphine and codeine doses are not available. Therefore, we investigated urine opiate pharmacokinetics after controlled oral administration of uncooked poppy seeds with known morphine and codeine content. Participants were administered two 45g oral poppy seed doses 8h apart, each containing 15.7mg morphine and 3mg codeine. Urine was collected ad libitum up to 32h after the first dose. Specimens were analyzed with the Roche Opiates II immunoassay at 2,000 and 300μg/L cutoffs, and the ThermoFisher CEDIA® Heroin Metabolite (6-acetylmorphine, 6AM) and Lin-Zhi 6AM immunoassays with 10μg/L cutoffs to determine if poppy seed ingestion could produce positive results in these heroin marker assays. In addition, all specimens were quantified for morphine and codeine by GC/MS. Participants (N=22) provided 391 urine specimens over 32h following dosing; 26.6% and 83.4% were positive for morphine at 2,000 and 300μg/L GC/MS cutoffs, respectively. For the 19 subjects who completed the study, morphine concentrations ranged from <300 to 7,522μg/L with a median peak concentration of 5,239μg/L. The median first morphine-positive urine sample at 2,000μg/L cutoff concentration occurred at 6.6h (1.2-12.1), with the last positive from 2.6 to 18h after the second dose. No specimens were positive for codeine at a cutoff concentration of 2,000μg/L, but 20.2% exceeded 300μg/L, with peak concentrations of 658 μg/L (284-1540). The Roche Opiates II immunoassay had efficiencies greater than 96% for the 2000 and 300μg/L cutoffs. The CEDIA 6AM immunoassay had a specificity of 91%, while the Lin-Zhi assay had no false positive results. These data provide valuable information for interpreting urine opiate results. PMID:24887324

  10. Stoichiometry of excreta and excretion rates of a stream-dwelling plethodontid salamander

    Data.gov (United States)

    U.S. Environmental Protection Agency — Stoichiometry of excreta and excretion rates of a stream-dwelling plethodontid salamander in Cincinnati, OH, USA. This dataset is associated with the following...

  11. Feed value of ensiled pig excreta, poultry litter or urea with ...

    African Journals Online (AJOL)

    Dnkosi

    2014-05-17

    May 17, 2014 ... Thus, pig excreta (PE) and PL are potential N sources for ruminant nutrition. ... litter or urea as N sources, and with molasses or bakery by-products as energy sources. ..... Kinetic parameters of ruminal degradation estimated.

  12. RESULTS OF THE EXCRETA BIOASSAY QUALITY CONTROL PROGRAM FOR APRIL 1, 2011 THROUGH MARCH 31, 2012

    Energy Technology Data Exchange (ETDEWEB)

    Antonio, Cheryl L.; MacLellan, Jay A.

    2012-10-31

    The performance statistics for the Hanford excreta radiobioassay program are presented and discussed. The performance period covers the second contract year of Contract 112512 - April 2011 to March 2012.

  13. Molecularly imprinted pipette-tip solid phase extraction for selective determination of fluoroquinolones in human urine using HPLC-DAD.

    Science.gov (United States)

    de Oliveira, Hanna Leijoto; da Silva Anacleto, Sara; da Silva, Anny Talita Maria; Pereira, Arnaldo César; de Souza Borges, Warley; Figueiredo, Eduardo Costa; Borges, Keyller Bastos

    2016-10-15

    A simple method using HPLC-DAD was developed for the determination of fluoroquinolones in human urine including ciprofloxacin (CIPRO), enrofloxacino (ENRO), marbofloxacino (MARBO) and norfloxacin (NOR). In addition, it was studied the extraction of fluoroquinolones in human urine samples using pipette tip-based molecularly imprinted polymers solid phase extraction (PT-MIPs-SPE). With the goal of finding the best procedure for extraction of four fluoroquinolones in human urine, several parameters that are likely to affect the efficiency of extraction during sample preparation, including the washing solvent, type and volume of eluent, amount of material, the volume of the sample, pH and the ionic strength were systematically optimized. Chromatographic separations of fluoroquinolones were hit within 10min using a Synergi(®) C18 (250×4.6mm, 4μm) column and mobile phase consisting of water (10mM of phosphoric acid, the pH adjusted at 3.29 with triethylamine) : acetonitrile (85.7: 14.3, v/v) at a flow rate of 1.5mLmin(-1). Detection was performed at 290nm. The average extraction recoveries/standard deviation relative to ENRO, CIPRO, NOR and MARBO were 96.40±5.51%, 42.47±4.81%, 41.82±7.99% and 87.49±4.70, respectively. The method was liner from 39 to 1260ngmL(-1) for each fluoroquinolone with correlation coefficient of 0.9904, 0.9910, 0.9914 and 0.9919, to ENRO, CIPRO, NOR and MARBO, respectively. The assays of within-day and between-day precision and accuracy for all analytes were studied at three concentration levels and were lower than 15%. The method was successfully employed in a preliminary cumulative urinary excretion study after administration of CIPRO to a healthy volunteer. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Methamphetamine and amphetamine isomer concentrations in human urine following controlled Vicks VapoInhaler administration.

    Science.gov (United States)

    Smith, Michael L; Nichols, Daniel C; Underwood, Paula; Fuller, Zachary; Moser, Matthew A; Flegel, Ron; Gorelick, David A; Newmeyer, Matthew N; Concheiro, Marta; Huestis, Marilyn A

    2014-10-01

    Legitimate use of legal intranasal decongestants containing l-methamphetamine may complicate interpretation of urine drug tests positive for amphetamines. Our study hypotheses were that commonly used immunoassays would produce no false-positive results and a recently developed enantiomer-specific gas chromatography-mass spectrometry (GC-MS) procedure would find no d-amphetamine or d-methamphetamine in urine following controlled Vicks VapoInhaler administration at manufacturer's recommended doses. To evaluate these hypotheses, 22 healthy adults were each administered one dose (two inhalations in each nostril) of a Vicks VapoInhaler every 2 h for 10 h on Day 1 (six doses), followed by a single dose on Day 2. Every urine specimen was collected as an individual void for 32 h after the first dose and assayed for d- and l-amphetamines specific isomers with a GC-MS method with >99% purity of R-(-)-α-methoxy-α-(trifluoromethyl)phenylacetyl derivatives and 10 µg/L lower limits of quantification. No d-methamphetamine or d-amphetamine was detected in any urine specimen by GC-MS. The median l-methamphetamine maximum concentration was 62.8 µg/L (range: 11.0-1,440). Only two subjects had detectable l-amphetamine, with maximum concentrations coinciding with l-methamphetamine peak levels, and always ≤ 4% of the parent's maximum. Three commercial immunoassays for amphetamines EMIT(®) II Plus, KIMS(®) II and DRI(®) had sensitivities, specificities and efficiencies of 100, 97.8, 97.8; 100, 99.6, 99.6 and 100, 100, 100%, respectively. The immunoassays had high efficiencies, but our first hypothesis was not affirmed. The EMIT(®) II Plus assay produced 2.2% false-positive results, requiring an enantiomer-specific confirmation.

  15. Selective fiber used for headspace solid-phase microextraction of abused drugs in human urine

    OpenAIRE

    Sunanta Wangkarn

    2007-01-01

    A sensitive and selective fiber for simultaneous analysis of three drugs of abuse (amphetamine, methamphetamine and ephedrine) in urine samples was explored using headspace solid phase microextraction and gas chromatography with flame ionization detection. Several parameters affecting extraction such as extraction time, extraction temperature, pH of solution and salt concentrations were investigated. Among five commercially available fibers, divinylbenzene/carboxen/ polydimethylsiloxane is th...

  16. Detection and characterization of prednisolone metabolites in human urine by LC-MS/MS.

    OpenAIRE

    Matabosch Geronès, Xavier; Pozo Mendoza, Óscar J., 1975-; Pérez Mañá, Clara; Papaseit Fontanet, Esther; Segura Noguera, Jordi; Ventura Alemany, Rosa

    2015-01-01

    Glucocorticosteroids are prohibited in sports when used by systemic administrations (e.g. oral), whereas they are allowed using other administration ways. Strategies to discriminate between administrations routes have to be developed by doping control laboratories. For this reason, the metabolism of prednisolone (PRED) was studied using liquid chromatography coupled to tandem mass spectrometry. A single oral (10 mg) dose of PRED was administered to two healthy male volunteers. Urine samples w...

  17. Structural Elucidation of a Carnosine-Acrolein Adduct and its Quantification in Human Urine Samples.

    Science.gov (United States)

    Bispo, Vanderson S; de Arruda Campos, Ivan P; Di Mascio, Paolo; Medeiros, Marisa H G

    2016-01-19

    Aldehydes accumulate in inflammation, during myocardial infarction and have been associated with pain symptoms. One pathway of aldehyde detoxification is the conjugation with carnosine. A 3-methylpyridinium carnosine adduct from the reaction of carnosine and acrolein was characterized using extensive spectroscopic measurements. The adduct with urinary concentrations of 1.82 ± 0.68 nmol/mg of creatinine is one of the most abundant acrolein metabolites in urine and opens promising therapeutic strategies for carnosine.

  18. Simultaneous determination of vanillylmandelic, homovanillic and 5-hydroxyindoleacetic acids in human urine by thin layer chromatography.

    Science.gov (United States)

    Alemany, G; Gamundí, A; Rosselló, C; Rial, R

    1996-01-01

    A TLC method for the simultaneous analysis of vanillylmandelic, homovanillic and 5-hydroxyindole-3-acetic acids in urine is described. The sample is cleaned up through a cyano minicolumn and extracted with diethyl ether. The acids are resolved by high-performance TLC, visualized by Folin Ciocalteau reagent and quantificated by densitometry at 600 nm with beta-(4-hydroxy-3-phenyl) acetic acid as the internal standard.

  19. Direct and label-free detection of the human growth hormone in urine by an ultrasensitive bimodal waveguide biosensor.

    Science.gov (United States)

    González-Guerrero, Ana Belén; Maldonado, Jesús; Dante, Stefania; Grajales, Daniel; Lechuga, Laura M

    2017-01-01

    A label-free interferometric transducer showing a theoretical detection limit for homogeneous sensing of 5 × 10(-8) RIU, being equivalent to a protein mass coverage resolution of 2.8 fg mm(-2) , is used to develop a high sensitive biosensor for protein detection. The extreme sensitivity of this transducer combined with a selective bioreceptor layer enables the direct evaluation of the human growth hormone (hGH) in undiluted urine matrix in the 10 pg mL(-1) range.

  20. GC-MS analysis of fluoxetine and its active metabolite norfluoxetine in human urine

    Directory of Open Access Journals (Sweden)

    Münevver Açıkkol

    2010-05-01

    Full Text Available A gas chromatographic-mass spectrometric (GC-MS method was developed for detection of fluoxetine and its active metabolite norfluoxetine in urine. Liquid and solid phase extraction were applied to urine samples using maprotiline as an internal standard (IS. The GC-MS analysis were carried out using HP-5MS capillary column. The linearity ranges of the method were 5-75 ng mL-1 for fluoxetine and 6-125 ng mL-1 for norfluoxetine by solid phase extraction (SPE, and 10-80 ng mL-1 for fluoxetine and also norfluoxetine by liquid- liquid extraction (LLE. Also the range of detection limits were between 1-10 ng mL-1, the range of quantification limits were between 5-10 ng mL-1 for fluoxetine and norfluoxetine by both SPE and LLE. The range of recoveries were between 87 -109 % by both SPE and LLE for analytes. The developed method allowed clinical and toxicological analysis of fluoxetine and norfluoxetine in urine samples.

  1. Liquid chromatographic determination of quinolones in water and human urine samples after microextraction by packed sorbent.

    Science.gov (United States)

    Rani, Susheela; Kumar, Ashwini; Malik, Ashok Kumar; Singh, Baldev

    2012-01-01

    A method for the simultaneous determination of quinolones in water and urine samples by microextraction in a sorbent-packed syringe (MEPS) with LC is described. MEPS is a new miniaturized SPE technique that can be used with chromatographic instruments without any modifications. In MEPS, approximately 1 mg of the solid packing material is inserted into a syringe (100-250 microL) as a plug. Sample preparation takes place on the packed bed. The new method is promising, easy to use, economical, and rapid. The determination of quinolones in groundwater and urine was performed using MEPS as a sample preparation method with LC-UV determination. Four quinolone antibiotics--enrofloxacin, enoxacin, danofloxacin, and nalidixic acid--in groundwater and urine samples were used as analytes. The extraction recovery was found to be between 64.9 and 98.9%. The results showed high correlation coefficients (R2 > 0.992) for all of the analytes within the calibration range. The LOQ was between 0.091 and 0.315 ng/mL.

  2. Mercury, lead and cadmium levels in the urine of 170 Spanish adults: a pilot human biomonitoring study.

    Science.gov (United States)

    Castaño, Argelia; Sánchez-Rodríguez, Jinny E; Cañas, Ana; Esteban, Marta; Navarro, Carmen; Rodríguez-García, Ana C; Arribas, Misericordia; Díaz, Gema; Jiménez-Guerrero, José A

    2012-02-01

    Human biomonitoring is a well-recognized tool for estimating the exposure of human populations to environmental pollutants. However, information regarding biomarker concentrations of many environmental chemicals in the general population is limited for many countries. The Spanish Environment Ministry has recently funded a human biomonitoring study on the Spanish general population. This study aims to determine reference levels for several biomarkers, especially heavy metals, persistent organic pollutants (POPs) and cotinine, in urine, whole blood, serum and hair, and will involve 2000 volunteers throughout Spain. Samples were taken during 2009-2010 and analyses are currently underway. The results presented herein were obtained in a pilot study carried out in the Madrid region. The study group comprised 170 volunteers, of which 79% were female and 21% male (age: 23-66 years). All participants were asked to complete a questionnaire regarding diet and living habits and provides a morning urine sample. The geometric means for total mercury (Hg), lead (Pb) and cadmium (Cd) were 1.23, 1.11 and 0.25 μg/g creatinine, respectively. Levels of Pb and Hg were higher than those reported for the general population in the USA and Germany, whereas Cd was in the same range (CDC, 2009; Becker et al., 2003). The values reported here are similar to those reported in other Spanish studies.

  3. Detection of Plasmodium vivax and Plasmodium falciparum DNA in human saliva and urine: loop-mediated isothermal amplification for malaria diagnosis.

    Science.gov (United States)

    Ghayour Najafabadi, Zahra; Oormazdi, Hormozd; Akhlaghi, Lame; Meamar, Ahmad Reza; Nateghpour, Mehdi; Farivar, Leila; Razmjou, Elham

    2014-08-01

    This study investigated loop-mediated isothermal amplification (LAMP) detection of Plasmodium falciparum and Plasmodium vivax in urine and saliva of malaria patients. From May to November 2011, 108 febrile patients referred to health centers in Sistan and Baluchestan Province of south-eastern Iran participated in the study. Saliva, urine, and blood samples were analyzed with nested PCR and LAMP targeting the species-specific nucleotide sequence of small subunit ribosomal RNA gene (18S rRNA) of P. falciparum and P. vivax and evaluated for diagnostic accuracy by comparison to blood nested PCR assay. When nested PCR of blood is used as standard, microscopy and nested PCR of saliva and urine samples showed sensitivity of 97.2%, 89.4% and 71% and specificity of 100%, 97.3% and 100%, respectively. LAMP sensitivity of blood, saliva, and urine was 95.8%, 47% and 29%, respectively, whereas LAMP specificity of these samples was 100%. Microscopy and nested PCR of saliva and LAMP of blood were comparable to nested PCR of blood (к=0.95, 0.83, and 0.94, respectively), but agreement for nested PCR of urine was moderate (к=0.64) and poor to fair for saliva LAMP and urine LAMP (к=0.38 and 0.23, respectively). LAMP assay showed low sensitivity for detection of Plasmodium DNA in human saliva and urine compared to results with blood and to nested PCR of blood, saliva, and urine. However, considering the advantages of LAMP technology and of saliva and urine sampling, further research into the method is worthwhile. LAMP protocol and precise preparation protocols need to be defined and optimized for template DNA of saliva and urine.

  4. Validation of a high performance liquid chromatography analysis for the determination of noradrenaline and adrenaline in human urine with an on-line sample purification

    DEFF Research Database (Denmark)

    Hansen, Åse Marie; Kristiansen, J; Nielsen, J L

    1999-01-01

    A high performance liquid chromatography (HPLC) method with fluorescence detection including an on-line purification was established for determination of catecholamines in human urine. The method was evaluated using samples of pooled urine spiked with catecholamines and validated for measurements...... deviation of the method was sufficiently low to distinguish between persons working with two different processes of garbage collection, i.e. collection using four wheeled containers versus collection using bins....

  5. Measuring the Safety of Excreta Disposal Behavior in India with the New Safe San Index: Reliability, Validity and Utility

    Directory of Open Access Journals (Sweden)

    Marion W. Jenkins

    2014-08-01

    Full Text Available Methods to assess household excreta disposal practices are critical for informing public health outcomes of efforts to improve sanitation in developing countries. We present a new metric, the Safe San Index (SSI, to quantify the hygienic safety of a household’s defecation and human feces disposal practices in India, where behavioral outcomes from on-going public expenditures to construct household sanitation facilities and eliminate open defecation are poorly measured. We define hygienic safety of feces disposal as capture in a hygienic sanitation facility. The SSI consists of 15 self-report items and two sub-scales, Latrine Use Frequency and Seven-Day Open Defecation Rate. Households are scored on a standardized scale from 0 (no defecation safely captured to 100 (all defecation safely captured. We present results of a pilot study in Odisha, India to apply the Index to assess excreta disposal behaviors among rural households and evaluate the reliability and validity of the Index for estimating the rate of correct and consistent sanitation facility usage of household with an improved latrine.

  6. Utilizing of Square Wave Voltammetry to Detect Flavonoids in the Presence of Human Urine

    Directory of Open Access Journals (Sweden)

    Rene Kizek

    2007-10-01

    Full Text Available About biological affecting of flavonoids on animal organisms is known less,thus we selected flavonoids, flavanones and flavones, and their glycosides, which wereexamined as potential inducers of cytochrome(s P450 when administrated by gavages intoexperimental male rats. The study was focused on induction of CYP1A1, the majorcytochrome P450 involved in carcinogen activation. The data obtained demonstrate thenecessity of taking into account not only ability of flavonoids to bind to Ah receptor(induction factor but also to concentrate on their distribution and metabolism (includingcolon microflora in the body. After that we examined certain flavonoids as potential inducers of cytochrome P450, we wanted to suggest and optimize suitable electrochemical technique for determination of selected flavonoids (quercetin, quercitrin, rutin, chrysin and diosmin in body liquids. For these purposes, we selected square wave voltannetry using carbon paste electrode. Primarily we aimed on investigation of their basic electrochemical behaviour. After that we have optimized frequency, step potential and supporting electrolyte. Based on the results obtained, we selected the most suitable conditions for determination of the flavonoids as follows: frequency 180 Hz, step potential 1.95 mV/s and phosphate buffer of pH 7 as supporting electrolyte. Detection limits (3 S/N of the flavonoids were from units to tens of nM except diosmin, where the limit were higher than μM. In addition, we attempted to suggest a sensor for analysis of flavonoids in urine. It clearly follows from the results obtained that flavonoids can be analysed in the presence of animal urine, because urine did not influence much the signals of flavonoids (recoveries of the signals were about 90 %.

  7. Gross alpha and beta activity analyses in urine-a routine laboratory method for internal human radioactivity detection.

    Science.gov (United States)

    Chen, Xiaowen; Zhao, Luqian; Qin, Hongran; Zhao, Meijia; Zhou, Yirui; Yang, Shuqiang; Su, Xu; Xu, Xiaohua

    2014-05-01

    The aim of this work was to develop a method to provide rapid results for humans with internal radioactive contamination. The authors hypothesized that valuable information could be obtained from gas proportional counter techniques by screening urine samples from potentially exposed individuals rapidly. Recommended gross alpha and beta activity screening methods generally employ gas proportional counting techniques. Based on International Standards Organization (ISO) methods, improvements were made in the evaporation process to develop a method to provide rapid results, adequate sensitivity, and minimum sample preparation and operator intervention for humans with internal radioactive contamination. The method described by an American National Standards Institute publication was used to calibrate the gas proportional counter, and urine samples from patients with or without radionuclide treatment were measured to validate the method. By improving the evaporation process, the time required to perform the assay was reduced dramatically. Compared with the reference data, the results of the validation samples were very satisfactory with respect to gross-alpha and gross-beta activities. The gas flow proportional counting method described here has the potential for radioactivity monitoring in the body. This method was easy, efficient, and fast, and its application is of great utility in determining whether a sample should be analyzed by a more complicated method, for example radiochemical and/or γ-spectroscopy. In the future, it may be used commonly in medical examination and nuclear emergency treatment.Health Phys. 106(5):000-000; 2014.

  8. Well-defined nanostructured surface-imprinted polymers for highly selective magnetic separation of fluoroquinolones in human urine.

    Science.gov (United States)

    He, Yonghuan; Huang, Yanyan; Jin, Yulong; Liu, Xiangjun; Liu, Guoquan; Zhao, Rui

    2014-06-25

    The construction of molecularly imprinted polymers on magnetic nanoparticles gives access to smart materials with dual functions of target recognition and magnetic separation. In this study, the superparamagnetic surface-molecularly imprinted nanoparticles were prepared via surface-initiated reversible addition-fragmentation chain transfer (RAFT) polymerization using ofloxacin (OFX) as template for the separation of fluoroquinolones (FQs). Benefiting from the living/controlled nature of RAFT reaction, distinct core-shell structure was successfully constructed. The highly uniform nanoscale MIP layer was homogeneously grafted on the surface of RAFT agent TTCA modified Fe3O4@SiO2 nanoparticles, which favors the fast mass transfer and rapid binding kinetics. The target binding assays demonstrate the desirable adsorption capacity and imprinting efficiency of Fe3O4@MIP. High selectivity of Fe3O4@MIP toward FQs (ofloxacin, pefloxacin, enrofloxacin, norfloxacin, and gatifloxacin) was exhibited by competitive binding assay. The Fe3O4@MIP nanoparticles were successfully applied for the direct enrichment of five FQs from human urine. The spiked human urine samples were determined and the recoveries ranging from 83.1 to 103.1% were obtained with RSD of 0.8-8.2% (n = 3). This work provides a versatile approach for the fabrication of well-defined MIP on nanomaterials for the analysis of complicated biosystems.

  9. Determination of 1-hydroxypyrene in human urine by high-performance liquid chromatography

    DEFF Research Database (Denmark)

    Hansen, Åse Marie; Poulsen, O M; Christensen, J M

    1993-01-01

    A high-performance liquid chromatography (HPLC)/fluorescence method for quantitative analysis of 1-hydroxypyrene in urine was developed. The method validation analysis showed the method to be in analytical control. No significant systematical errors could be demonstrated. The entire run time...... of chromatography was 10 min using isocratic elution (acetonitrile-water, 70:30), and the retention time for 1-hydroxypyrene was 3.5 min. The short run time in combination with the low limit detection (1.37 nmol/L) makes the method potentially applicable for surveillance of pyrene exposure in work environments...

  10. Widespread occurrence of perchlorate in water, foodstuffs and human urine collected from Kuwait and its contribution to human exposure.

    Science.gov (United States)

    Alomirah, Husam F; Al-Zenki, Sameer F; Alaswad, Marivi C; Alruwaih, Noor A; Wu, Qian; Kannan, Kurunthachalam

    2016-06-01

    Perchlorate is a thyroid hormone-disrupting compound and is reported to occur widely in the environment. Little is known on human exposure to perchlorate in Kuwait. In this study, 218 water samples, 618 commonly consumed foodstuffs and 532 urine samples collected from Kuwait were analysed to assess the exposure of the Kuwaiti population to perchlorate. For the estimation of daily intake of perchlorate, food consumption rates were obtained from the National Nutrition Survey in the State of Kuwait (NNSSK). The results showed that leafy vegetables accounted for a major share of perchlorate exposure among the Kuwaiti population at 0.062 µg kg(-)(1) bw day(-)(1) (36.2%), followed by fruits at 0.026 µg kg(-)(1) bw day(-)(1) (15.3%) and non-leafy vegetables at 0.017 µg kg(-)(1) bw day(-)(1) (10.1%). The urinary perchlorate geometric mean (GM) concentrations ranged from 8.51 to 17.1 µg l(-)(1) for the five age groups, which were higher than those reported in other countries. The estimated urinary perchlorate exposure for the Kuwaiti general population was 0.42 µg kg(-)(1) bw day(-)(1), which was higher than that reported for the United States. The dietary intake of perchlorate for the Kuwaiti population ranged from 0.14 to 0.67 µg kg(-)(1) bw day(-)(1) for the five age groups, with a mean total daily intake of 0.17 µg kg(-)(1) bw day(-)(1) for the general population. The highest estimated dietary mean daily intake of perchlorate (0.67 µg kg(-)(1) bw day(-)(1)) was found for children at 3-5 years. The estimated dietary perchlorate exposure in Kuwait is higher than the recommended mean reference dose (RfD) but lower than that of provisional maximum tolerable daily intake (PMTDI) set by the Joint FAO/WHO Expert Committee on Food Additives (JECFA).

  11. Sunscreens in human plasma and urine after repeated whole-body topical application

    DEFF Research Database (Denmark)

    Janjua, N.R.; Kongshoj, B.; Andersson, A.M.;

    2008-01-01

    Background The three chemical ultraviolet absorbers benzophenone-3 (BP-3), octyl-methoxycinnamate (OMC) and 3-(4-methylbenzylidene) camphor (4-MBC) are commercially used in sunscreens worldwide. Apart from sun protection, they may possess endocrine-disrupting effects in animals and in vitro...... the first application, all three sunscreens were detectable in plasma. The maximum median plasma concentrations were 187 ng/mL BP-3, 16 ng/mL 4-MBC and 7 ng/mL OMC for females and 238 ng/mL BP-3, 18 ng/mL 4-MBC and 16 ng/mL OMC for men. In the females, urine levels of 44 ng/mL BP-3 and 4 ng/mL of 4-MBC...... and 6 ng/mL OMC were found, and in the males, urine levels of 81 ng/mL BP-3, 4 ng/mL of 4-MBC and OMC were found. In plasma, the 96-h median concentrations were higher compared with the 24-h concentrations for 4-MBC and OMC in men and for BP-3 and 4-MBC in females Udgivelsesdato: 2008/4...

  12. Selective fiber used for headspace solid-phase microextraction of abused drugs in human urine

    Directory of Open Access Journals (Sweden)

    Sunanta Wangkarn

    2007-09-01

    Full Text Available A sensitive and selective fiber for simultaneous analysis of three drugs of abuse (amphetamine, methamphetamine and ephedrine in urine samples was explored using headspace solid phase microextraction and gas chromatography with flame ionization detection. Several parameters affecting extraction such as extraction time, extraction temperature, pH of solution and salt concentrations were investigated. Among five commercially available fibers, divinylbenzene/carboxen/ polydimethylsiloxane is the most sensitive and selective fiber at pH 10.0, extraction temperature at 80 C for 20 min and desorption temperature at 220 C for 2 min. Under the optimal conditions, the proposed solid phase microextraction method provided good linearity in the ranges 0.1-10 µg/ml for amphetamine and methamphetamine and 0.5-20 µg/ml for ephedrine. The detection limits for amphetamine, methamphetamine and ephedrine were 9, 3 and 30 ng/ml, respectively. The recoveries of three drugs in urine samples were exceeding 85%.

  13. Urine Odor

    Science.gov (United States)

    ... References Brunzel NA. Physical examination of urine. In: Fundamentals of Urine and Body Fluid Analysis. 3rd ed. St. Louis, Mo.: Saunders Elsevier; 2013:97. McPherson RA, et al., eds. Henry's Clinical Diagnosis and Management by Laboratory Methods. 23rd ed. St. Louis, Mo.: ...

  14. Metabolism studies of the Kratom alkaloid speciociliatine, a diastereomer of the main alkaloid mitragynine, in rat and human urine using liquid chromatography-linear ion trap mass spectrometry.

    Science.gov (United States)

    Philipp, Anika A; Wissenbach, Dirk K; Weber, Armin A; Zapp, Josef; Maurer, Hans H

    2011-03-01

    Mitragyna speciosa (Kratom) is currently used as a drug of abuse. When monitoring its abuse in urine, several alkaloids and their metabolites must be considered. In former studies, mitragynine (MG), its diastereomer speciogynine (SG), and paynantheine and their metabolites could be identified in rat and human urine using LC-MS(n). In Kratom users' urines, besides MG and SG, further isomeric compounds were detected. To elucidate whether the MG and SG diastereomer speciociliatine (SC) and its metabolites represent further compounds, the phase I and II metabolites of SC were identified first in rat urine after the administration of the pure alkaloid. Then, the identified rat metabolites were screened for in the urine of Kratom users using the above-mentioned LC-MS(n) procedure. Considering the mass spectra and retention times, it could be confirmed that SC and its metabolites are so far the unidentified isomers in human urine. In conclusion, SC and its metabolites can be used as further markers for Kratom use, especially by consumption of raw material or products that contain a high amount of fruits of the Malaysian plant M. speciosa.

  15. Direct determination of uranium in human urine by Icp-SFMS; Determinacion directa de uranio en orina humana por ICP-SFMS

    Energy Technology Data Exchange (ETDEWEB)

    Hernandez M, H. [ININ, Departamento de Quimica, Carretera Mexico-Toluca s/n, 52750 Ocoyoacac, Estado de Mexico (Mexico); Yllera de Ll, A., E-mail: hector.hernandez520@gmail.com [Centro de Investigaciones Energeticas, Medioambientales y Tecnologicas, Departamento de Medio Ambiente, Av. Complutense 22, 28040 Madrid (Spain)

    2013-10-15

    The success of the measurement and the evaluation of the internal exposure are highly dependent of the effective capacities for the radiation measurement in biological samples (mainly urine and the feces). Usually, during the samples bioassay of human urine, a pre-concentration and purification of the radionuclides is carried out previously to the quantitative analysis. These stages, as the analysis time are the main source of uncertainty in the measurement process. In the uranium case, this is not necessary when are used mass spectrometry techniques, in particular, Mass Spectrometry of Magnetic Sector with Inductively Coupled Plasma (Icp-SFMS). This work presents the results obtained for the uranium analysis in samples of human urine during the participation in the inter-comparison exercises of the Association pour la Promotion de Controle de Qualite des Analyses de Biologie Medicale en Radiotoxicologie (PROCORAD) in the period 2010 and 2011. The analyses were realized directly in the diluted urine samples (dilution factor 1:20) in 5% of HNO{sub 3}. The obtained results, were normalized to the total urine sample (V = 0.5 L), these values coincide with the waited reference values of uranium in the urine sample. Additionally, were calculated the detection limits of {sup 235}U= 0.049 x 10{sup -3} μg L{sup -1} and {sup 238}U= 7.37 x 10{sup -3} μg L{sup -1}. (author)

  16. A Population Pharmacokinetic Model for Disposition in Plasma, Saliva and Urine of Scopolamine after Intranasal Administration to Healthy Human Subjects

    Science.gov (United States)

    Wu, L.; Tam, V. H.; Chow, D. S. L.; Putcha, L.

    2014-01-01

    An intranasal gel formulation of scopolamine (INSCOP) was developed for the treatment of Space Motion Sickness. The bioavailability and pharmacokinetics (PK) were evaluated under the Food and Drug Administration guidelines for clinical trials with an Investigative New Drug (IND) protocol. The aim of this project was to develop a PK model that can predict the relationship between plasma, saliva and urinary scopolamine concentrations using data collected from the IND clinical trials with INSCOP. Methods: Twelve healthy human subjects were administered three dose levels (0.1, 0.2 and 0.4 mg) of INSCOP. Serial blood, saliva and urine samples were collected between 5 min and 24 h after dosing and scopolamine concentrations were measured by using a validated LC-MS-MS assay. Pharmacokinetic Compartmental models, using actual dosing and sampling times, were built using Phoenix (version 1.2). Model selection was based on the likelihood ratio test on the difference of criteria (-2LL) and comparison of the quality of fit plots. Results: The best structural model for INSCOP (minimal -2LL= 502.8) was established. It consisted of one compartment each for plasma, saliva and urine, respectively, which were connected with linear transport processes except the nonlinear PK process from plasma to saliva compartment. The best-fit estimates of PK parameters from individual PK compartmental analysis and Population PK model analysis were shown in Tables 1 and 2, respectively. Conclusion: A population PK model that could predict population and individual PK of scopolamine in plasma, saliva and urine after dosing was developed and validated. Incorporating a non-linear transfer from plasma to saliva compartments resulted in a significantly improved model fitting. The model could be used to predict scopolamine plasma concentrations from salivary and urinary drug levels, allowing non-invasive therapeutic monitoring of scopolamine in space and other remote environments.

  17. Quantification of clenbuterol at trace level in human urine by ultra-high pressure liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Nicoli, Raul; Petrou, Michael; Badoud, Flavia; Dvorak, Jiri; Saugy, Martial; Baume, Norbert

    2013-05-31

    Clenbuterol is a β2 agonist agent with anabolic properties given by the increase in the muscular mass in parallel to the decrease of the body fat. For this reason, the use of clenbuterol is forbidden by the World Anti-Doping Agency (WADA) in the practice of sport. This compound is of particular interest for anti-doping authorities and WADA-accredited laboratories due to the recent reporting of risk of unintentional doping following the eating of meat contaminated with traces of clenbuterol in some countries. In this work, the development and the validation of an ultra-high pressure liquid chromatography coupled to electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) method for the quantification of clenbuterol in human urine is described. The analyte was extracted from urine samples by liquid-liquid extraction (LLE) in basic conditions using tert butyl-methyl ether (TBME) and analyzed by UHPLC-MS/MS with a linear gradient of acetonitrile in 9min only. The simple and rapid method presented here was validated in compliance with authority guidelines and showed a limit of quantification at 5pg/mL and a linearity range from 5pg/mL to 300pg/mL. Good trueness (85.8-105%), repeatability (5.7-10.6% RSD) and intermediate precision (5.9-14.9% RSD) results were obtained. The method was then applied to real samples from eighteen volunteers collecting urines after single oral doses administration (1, 5 and 10μg) of clenbuterol-enriched yogurts.

  18. Phase I metabolism of the highly potent synthetic cannabinoid MDMB-CHMICA and detection in human urine samples.

    Science.gov (United States)

    Franz, Florian; Angerer, Verena; Moosmann, Bjoern; Auwärter, Volker

    2017-05-01

    Among the recently emerged synthetic cannabinoids, MDMB-CHMICA (methyl N-{[1-(cyclohexylmethyl)-1H-indol-3-yl]carbonyl}-3-methylvalinate) shows an extraordinarily high prevalence in intoxication cases, necessitating analytical methods capable of detecting drug uptake. In this study, the in vivo phase I metabolism of MDMB-CHMICA was investigated using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) and liquid chromatography-electrospray ionization-quadrupole time-of-flight-mass spectrometry (LC-ESI-Q ToF-MS) techniques. The main metabolites are formed by hydrolysis of the methyl ester and oxidation of the cyclohexyl methyl side chain. One monohydroxylated metabolite, the ester hydrolysis product and two further hydroxylated metabolites of the ester hydrolysis product are suggested as suitable targets for a selective and sensitive detection in urine. All detected in vivo metabolites could be verified in vitro using a human liver microsome assay. Two of the postulated main metabolites were successfully included in a comprehensive LC-ESI-MS/MS screening method for synthetic cannabinoid metabolites. The screening of 5717 authentic urine samples resulted in 818 cases of confirmed MDMB-CHMICA consumption (14%). Since the most common route of administration is smoking, smoke condensates were analyzed to identify relevant thermal degradation products. Pyrolytic cleavage of the methyl ester and amide bond led to degradation products which were also formed metabolically. This is particularly important in hair analysis, where detection of metabolites is commonly considered a proof of consumption. In addition, intrinsic activity of MDMB-CHMICA at the CB1 receptor was determined applying a cAMP accumulation assay and showed that the compound is a potent full agonist. Based on the collected data, an enhanced interpretation of analytical findings in urine and hair is facilitated. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John

  19. Classical Molecular Tests Using Urine Samples as a Potential Screening Tool for Human Papillomavirus Detection in Human Immunodeficiency Virus-Infected Women

    OpenAIRE

    Munoz, Marina; Camargo, Milena; Soto-De Leon, Sara C.; Sanchez, Ricardo; Pineda-Peña, Andrea C.; Perez-Prados, Antonio; Patarroyo, Manuel E.; Patarroyo, Manuel A.

    2013-01-01

    Human papillomavirus (HPV) is the main risk factor associated with the development of cervical cancer (CC); however, there are other factors, such as immunosuppression caused by the human immunodeficiency virus (HIV), that favor progression of the illness. This study was thus aimed at evaluating the functionality of classical PCR-based molecular tests for the generic identification of HPV DNA (GP5+/GP6+, MY09/MY11, and pU1M/2R primers, individually or in combination) using cervical and urine ...

  20. Effects of probiotic supplementation in different energy and nutrient density diets on performance, egg quality, excreta microflora, excreta noxious gas emission, and serum cholesterol concentrations in laying hens.

    Science.gov (United States)

    Zhang, Z F; Kim, I H

    2013-10-01

    This 6-wk study was conducted to determine the effects of probiotic (Enterococcus faecium DSM 7134) supplementation of different energy and nutrient density diets on performance, egg quality, excreta microflora, excreta noxious gas emission, and serum cholesterol concentrations in laying hens. A total of 432 Hy-Line brown layers (40 wk old) were allotted into 4 dietary treatments with 2 levels of probiotic supplementation (0 or 0.01%) and 2 levels of energy (2,700 or 2,800 kcal ME/kg) and nutrient density. Weekly feed intake, egg quality, and daily egg production were determined. Eighteen layers per treatment (2 layers/replication) were bled to determine serum cholesterol concentrations at wk 3 and 6. Excreta microbial shedding of Lactobacillus, Escherichia coli, and Salmonella and noxious gas emission were determined at the end of the experiment. Hens fed the high-energy and high-nutrient-density diets had less (P hens fed the diets supplemented with the probiotic had greater (P hens fed the diets without the probiotic. Dietary supplementation of the probiotic increased (P = 0.01) excreta Lactobacillus counts and decreased (P = 0.02) Escherichia coli counts compared with hens fed the diets without the probiotic. The excreta ammonia emission was decreased (P = 0.02) in hens fed the probiotic diets compared with hens fed the diets without the probiotic. Serum total cholesterol concentration was decreased (P hens with the probiotic at wk 3 and 6. Layers fed the probiotic-incorporated diets had greater (P hens fed the nonsupplemented diets at wk 6. Interactive effects (P hens.

  1. Sensitive and cost-effective LC-MS/MS method for quantitation of CVT-6883 in human urine using sodium dodecylbenzenesulfonate additive to eliminate adsorptive losses.

    Science.gov (United States)

    Chen, Chungwen; Bajpai, Lakshmikant; Mollova, Nevena; Leung, Kwan

    2009-04-01

    CVT-6883, a novel selective A(2B) adenosine receptor antagonist currently under clinical development, is highly lipophilic and exhibits high affinity for non-specific binding to container surfaces, resulting in very low recovery in urine assays. Our study showed the use of sodium dodecylbenzenesulfonate (SDBS), a low-cost additive, eliminated non-specific binding problems in the analysis of CVT-6883 in human urine without compromising sensitivity. A new sensitive and selective LC-MS/MS method for quantitation of CVT-6883 in the range of 0.200-80.0ng/mL using SDBS additive was therefore developed and validated for the analysis of human urine samples. The recoveries during sample collection, handling and extraction for the analyte and internal standard (d(5)-CVT-6883) were higher than 87%. CVT-6883 was found stable under the following conditions: in extract - at ambient temperature for 3 days, under refrigeration (5 degrees C) for 6 days; in human urine (containing 4mM SDBS) - after three freeze/thaw cycles, at ambient temperature for 26h, under refrigeration (5 degrees C) for 94h, and in a freezer set to -20 degrees C for at least 2 months. The results demonstrated that the validated method is sufficiently sensitive, specific, and cost-effective for the analysis of CVT-6883 in human urine and will provide a powerful tool to support the clinical programs for CVT-6883.

  2. On-line clean-up and screening of oxacillin and cloxacillin in human urine and plasma with a weak ion exchange monolithic column.

    Science.gov (United States)

    Yang, Gengliang; Feng, Sha; Liu, Haiyan; Yin, Junfa; Zhang, Li; Cai, Liping

    2007-07-01

    A weak ion exchange monolithic column prepared by modifying the GMA-MAA-EDMA (glycidyl methacrylate-methacrylic acid-ethylene glycol dimethacrylate) monoliths with ethylenediamine was applied to remove matrix compounds in biological fluid. Using this monolithic column, on-line clean-up and screening of oxacillin and cloxacillin in human urine and plasma samples had been investigated. Chromatography was performed by reversed-phase HPLC on a C(18) column with ultraviolet detection at 225 nm. Results showed that the ion exchange monolithic column could be used for deproteinization and retaining oxacillin and cloxacillin in human urine and plasma, which provided a simple and fast method for assaying drugs in human urine and plasma.

  3. ASSOCIATION OF DIARRHOEA WITH PRACTICES OF HAND WASHING AND EXCRETA DISPOSAL IN CHILDREN

    Directory of Open Access Journals (Sweden)

    Avinash Kr.

    2015-04-01

    Full Text Available INTRODUCTION: Diarrhoea is a common and preventable disease, but still it accounts for approximately 11% of all mortality in children under five years of age. Diarrhoeal infections are more common when there is a shortage of adequate sanitation, hygiene and safe water f or drinking, cooking and cleaning. To prevent stool pathogens from gaining access to the domestic environment, efforts should concentrate on hand washing after stool contact, especially after defaecation or after cleaning up a child. MATERIALS AND METHODS: Present study was conducted from Dec - 2014 to Feb - 2015, after selecting a total of 500 children. The association of diarrhoea in these children were studied in relation practices of hand washing and excreta disposal among them. OBSERVATION: It was observed in our study that the incidence of diarrhoea was more in children excreting inside the household premises (32%, in comparison to those excreting outside (11.5%. Incidence of diarrhoea in children having the habits of washing hands was lower (13.6%, tha n the children not having the habit (21.8%. CONCLUSION: Based on the above findings we conclude that human stools in the domestic environment are a source of diarrhoeal infection, and safe disposal of stools should be one of the key measures to prevent di arrhoeal diseases.

  4. Multi-residue analysis of eight thioamphetamine designer drugs in human urine by liquid chromatography/tandem mass spectrometry.

    Science.gov (United States)

    Nieddu, Maria; Boatto, Gianpiero; Pirisi, Maria Antonietta; Baralla, Elena

    2009-10-01

    An analytical procedure for the simultaneous determination in human urine of several thioamphetamine designer drugs (2C-T and ALEPH series) is reported. The quantitative analysis was performed by liquid chromatography/tandem mass spectrometry and has been fully validated. The mass spectrometer was operated in positive-ion, selected reaction monitoring (SRM) mode. In order to minimize interferences with matrix components and to preconcentrate target analytes, solid-phase extraction was introduced in the method as a clean-up step. The entire method was validated for selectivity, linearity, precision and accuracy. The method turned out to be specific, sensitive, and reliable for the analysis of amphetamine derivatives in urine samples. The calibration curves were linear over the concentration range of 1 to 100 ng mL(-1) for all drugs with correlation coefficients that exceeded 0.996. The lower limits of detection (LODs) and quantification (LOQs) ranged from 1.2 to 4.9 ng mL(-1) and from 3.2 to 9.6 ng mL(-1), respectively.

  5. In-Depth Characterization and Validation of Human Urine Metabolomes Reveal Novel Metabolic Signatures of Lower Urinary Tract Symptoms

    Science.gov (United States)

    Hao, Ling; Greer, Tyler; Page, David; Shi, Yatao; Vezina, Chad M.; Macoska, Jill A.; Marker, Paul C.; Bjorling, Dale E.; Bushman, Wade; Ricke, William A.; Li, Lingjun

    2016-08-01

    Lower urinary tract symptoms (LUTS) are a range of irritative or obstructive symptoms that commonly afflict aging population. The diagnosis is mostly based on patient-reported symptoms, and current medication often fails to completely eliminate these symptoms. There is a pressing need for objective non-invasive approaches to measure symptoms and understand disease mechanisms. We developed an in-depth workflow combining urine metabolomics analysis and machine learning bioinformatics to characterize metabolic alterations and support objective diagnosis of LUTS. Machine learning feature selection and statistical tests were combined to identify candidate biomarkers, which were statistically validated with leave-one-patient-out cross-validation and absolutely quantified by selected reaction monitoring assay. Receiver operating characteristic analysis showed highly-accurate prediction power of candidate biomarkers to stratify patients into disease or non-diseased categories. The key metabolites and pathways may be possibly correlated with smooth muscle tone changes, increased collagen content, and inflammation, which have been identified as potential contributors to urinary dysfunction in humans and rodents. Periurethral tissue staining revealed a significant increase in collagen content and tissue stiffness in men with LUTS. Together, our study provides the first characterization and validation of LUTS urinary metabolites and pathways to support the future development of a urine-based diagnostic test for LUTS.

  6. Effects of Gentle Human Touch and Field Massage on Urine Cortisol Level in Premature Infants: A Randomized, Controlled Clinical Trial

    Science.gov (United States)

    Asadollahi, Malihe; Jabraeili, Mahnaz; Mahallei, Majid; Asgari Jafarabadi, Mohammad; Ebrahimi, Sakine

    2016-01-01

    Introduction: Hospitalization in neonatal intensive care unit may leads to many stresses for premature infants. Since premature infants cannot properly process stressors, identifying interventions that reduce the stress level for them is seems necessary. The aim of present study was to compare the effects of Field massage and Gentle Human Touch (GHT) techniques on the urine level of cortisol, as an indicator of stress in preterm infants. Methods: This randomized, controlled clinical trial was carried out in Al-Zahra hospital, Tabriz. A total of 84 premature infants were randomly assigned into three groups. First groups were touched by their mothers three times a day (15 minutes in each session) for 5 days by GHT technique. The second group was received 15 minutes Field massage with sunflower oil three times a day by their mothers for 5 days. The third group received routine care. In all groups, 24-hours urine samples were collected in the first and sixth day after the intervention and analyzed for cortisol level. Data were analyzed by SPSS software. Results: There were significant differences between mean of changes in cortisol level between GHT and control groups and Field massage and control groups (0.026). Conclusion: Although the massage with Field technique resulted in a significant reduction in blood cortisol level, but the GHT technique have also a similar effect. So, both methods are recommended for decreasing of stress in preterm infants. PMID:27752484

  7. Urine-sample-derived human induced pluripotent stem cells as a model to study PCSK9-mediated autosomal dominant hypercholesterolemia.

    Science.gov (United States)

    Si-Tayeb, Karim; Idriss, Salam; Champon, Benoite; Caillaud, Amandine; Pichelin, Matthieu; Arnaud, Lucie; Lemarchand, Patricia; Le May, Cédric; Zibara, Kazem; Cariou, Bertrand

    2016-01-01

    Proprotein convertase subtilisin kexin type 9 (PCSK9) is a critical modulator of cholesterol homeostasis. Whereas PCSK9 gain-of-function (GOF) mutations are associated with autosomal dominant hypercholesterolemia (ADH) and premature atherosclerosis, PCSK9 loss-of-function (LOF) mutations have a cardio-protective effect and in some cases can lead to familial hypobetalipoproteinemia (FHBL). However, limitations of the currently available cellular models preclude deciphering the consequences of PCSK9 mutation further. We aimed to validate urine-sample-derived human induced pluripotent stem cells (UhiPSCs) as an appropriate tool to model PCSK9-mediated ADH and FHBL. To achieve our goal, urine-sample-derived somatic cells were reprogrammed into hiPSCs by using episomal vectors. UhiPSC were efficiently differentiated into hepatocyte-like cells (HLCs). Compared to control cells, cells originally derived from an individual with ADH (HLC-S127R) secreted less PCSK9 in the media (-38.5%; P=0.038) and had a 71% decrease (Pcells originally derived from an individual with FHBL (HLC-R104C/V114A) displayed a strong decrease in PCSK9 secretion (-89.7%; Pcells for reprogramming and hepatocyte differentiation, but also a powerful tool to further decipher PCSK9 mutations and function.

  8. Analysis of the anti-Parkinson drug pramipexole in human urine by capillary electrophoresis with laser-induced fluorescence detection.

    Science.gov (United States)

    Musenga, Alessandro; Kenndler, Ernst; Morganti, Emanuele; Rasi, Fabrizio; Raggi, Maria Augusta

    2008-09-19

    A sensitive method based on capillary electrophoresis with laser-induced fluorescence detection has been developed for the analysis of the non-ergoline dopamine agonist pramipexole in human urine. Separation was carried out in uncoated fused silica capillaries (75microm internal diameter, 75.0 and 60.0cm total and effective length, respectively), with a background electrolyte composed of borate buffer (50mM, pH 10.3), tetrabutylammonium bromide (30mM), and acetone (15%, v/v). Applying a 20kV voltage, the electrophoretic run is completed within 12min. A sample pre-treatment procedure based on liquid/liquid extraction with ethyl acetate, followed by derivatisation of pramipexole with fluorescein isothiocyanate at pH 9, allows the complete removal of biological interferences, with extraction yields always higher than 94.5%. Method validation gave good linearity (r(2)=0.9992) in the 25.0-1000ngmL(-1) range; limit of detection and limit of quantitation were 10.0 and 25.0ngmL(-1), respectively; precision was 90.0. The method was applied to the analysis of urine samples from patients undergoing therapy with pramipexole.

  9. New Potential Biomarker for Methasterone Misuse in Human Urine by Liquid Chromatography Quadrupole Time of Flight Mass Spectrometry

    Science.gov (United States)

    Zhang, Jianli; Lu, Jianghai; Wu, Yun; Wang, Xiaobing; Xu, Youxuan; Zhang, Yinong; Wang, Yan

    2016-01-01

    In this study, methasterone urinary metabolic profiles were investigated by liquid chromatography quadrupole time of flight mass spectrometry (LC-QTOF-MS) in full scan and targeted MS/MS modes with accurate mass measurement. A healthy male volunteer was asked to take the drug and liquid–liquid extraction was employed to process urine samples. Chromatographic peaks for potential metabolites were hunted out with the theoretical [M − H]− as a target ion in a full scan experiment and actual deprotonated ions were studied in targeted MS/MS experiment. Fifteen metabolites including two new sulfates (S1 and S2), three glucuronide conjugates (G2, G6 and G7), and three free metabolites (M2, M4 and M6) were detected for methasterone. Three metabolites involving G4, G5 and M5 were obtained for the first time in human urine samples. Owing to the absence of helpful fragments to elucidate the steroid ring structure of methasterone phase II metabolites, gas chromatography mass spectrometry (GC-MS) was employed to obtain structural information of the trimethylsilylated phase I metabolite released after enzymatic hydrolysis and the potential structure was inferred using a combined MS method. Metabolite detection times were also analyzed and G2 (18-nor-17β-hydroxymethyl-2α, 17α-dimethyl-androst-13-en-3α-ol-ξ-O-glucuronide) was thought to be new potential biomarker for methasterone misuse which can be detected up to 10 days. PMID:27669235

  10. Human Urine Proteomics: Analytical Techniques and Clinical Applications in Renal Diseases

    Directory of Open Access Journals (Sweden)

    Shiva Kalantari

    2015-01-01

    Full Text Available Urine has been in the center of attention among scientists of clinical proteomics in the past decade, because it is valuable source of proteins and peptides with a relative stable composition and easy to collect in large and repeated quantities with a noninvasive procedure. In this review, we discuss technical aspects of urinary proteomics in detail, including sample preparation, proteomic technologies, and their advantage and disadvantages. Several recent experiments are presented which applied urinary proteome for biomarker discovery in renal diseases including diabetic nephropathy, immunoglobulin A (IgA nephropathy, focal segmental glomerulosclerosis, lupus nephritis, membranous nephropathy, and acute kidney injury. In addition, several available databases in urinary proteomics are also briefly introduced.

  11. Ion-exchange selectivity of diclofenac, ibuprofen, ketoprofen, and naproxen in ureolyzed human urine.

    Science.gov (United States)

    Landry, Kelly A; Sun, Peizhe; Huang, Ching-Hua; Boyer, Treavor H

    2015-01-01

    This research advances the knowledge of ion-exchange of four non-steroidal anti-inflammatory drugs (NSAIDs) - diclofenac (DCF), ibuprofen (IBP), ketoprofen (KTP), and naproxen (NPX) - and one analgesic drug-paracetamol (PCM) - by strong-base anion exchange resin (AER) in synthetic ureolyzed urine. Freundlich, Langmuir, Dubinin-Astakhov, and Dubinin-Radushkevich isotherm models were fit to experimental equilibrium data using nonlinear least squares method. Favorable ion-exchange was observed for DCF, KTP, and NPX, whereas unfavorable ion-exchange was observed for IBP and PCM. The ion-exchange selectivity of the AER was enhanced by van der Waals interactions between the pharmaceutical and AER as well as the hydrophobicity of the pharmaceutical. For instance, the high selectivity of the AER for DCF was due to the combination of Coulombic interactions between quaternary ammonium functional group of resin and carboxylate functional group of DCF, van der Waals interactions between polystyrene resin matrix and benzene rings of DCF, and possibly hydrogen bonding between dimethylethanol amine functional group side chain and carboxylate and amine functional groups of DCF. Based on analysis of covariance, the presence of multiple pharmaceuticals did not have a significant effect on ion-exchange removal when the NSAIDs were combined in solution. The AER reached saturation of the pharmaceuticals in a continuous-flow column at varying bed volumes following a decreasing order of DCF > NPX ≈ KTP > IBP. Complete regeneration of the column was achieved using a 5% (m/m) NaCl, equal-volume water-methanol solution. Results from multiple treatment and regeneration cycles provide insight into the practical application of pharmaceutical ion-exchange in ureolyzed urine using AER.

  12. Determination of Sparfloxacin in Human Urine by Reversed-Phase High Performance Liquid Chromatography With Nitrous Acid and Hydroiodic Pre-Column Derivatization

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Sparfloxacin can be oxidized by nitrous acid, then react with hydroiodic acid to form a fluorescent derivative. Based on this, a reversed-phase high performance liquid chromatographic pre-column derivatization new method is described for the determination of sparfloxacin in human urine. The linear range is 0.05 mg/L to 4.0 mg/L, the recoveries are 91.5%~95.7% and the RSD is 1.2%~4.2%. The results showed that this method is suitable for the determination of sparfloxacin in human urine.

  13. Storage stability studies for tributyltin determination in human urine samples using headspace solid-phase microextraction and gas chromatography mass spectrometry.

    Science.gov (United States)

    Zachariadis, G A; Tzollas, N M; Nikolaou, M; Rosenberg, E

    2013-03-01

    A headspace solid-phase micro-extraction (HS-SPME) method was employed in order to study the effect of storage conditions of human urine samples spiked with tributyltin (TBT) using gas chromatography and mass spectrometry. To render the analyte more volatile, the derivatization (ethylation) was made in situ by sodium tetraethylborate (NaBEt(4) ), which was added directly to dilute unpreserved urine samples and in buffers of similar acidity. The stability of TBT in human urine matrix was compared with the stability of TBT in buffer solutions of similar pH value. Critical parameters of storage conditions such as temperature and time, which affect the stability of TBT in this kind of matrix, were examined extensively. The tests showed that the stability of TBT remains practically satisfactory for a maximum of 2 days of storage either at +4 or 20°C. Greater variations were observed in the concentration of TBT in human urine samples at +4°C and lower ones at -20°C over a month's storage. The freeze-thaw cycles have negative effect on the stability and should be kept to a minimum. The results from spiked urine samples are also discussed in comparison to those acquired from buffer solutions of equal TBT concentration.

  14. Evaluation of the in vitro growth of urinary tract infection-causing gram-negative and gram-positive bacteria in a proposed synthetic human urine (SHU) medium.

    Science.gov (United States)

    Ipe, Deepak S; Ulett, Glen C

    2016-08-01

    Bacteriuria is a hallmark of urinary tract infection (UTI) and asymptomatic bacteriuria (ABU), which are among the most frequent infections in humans. A variety of gram-negative and gram-positive bacteria are associated with these infections but Escherichia coli contributes up to 80% of cases. Multiple bacterial species including E. coli can grow in human urine as a means to maintain colonization during infections. In vitro bacteriuria studies aimed at modeling microbial growth in urine have utilized various compositions of synthetic human urine (SHU) and a Composite SHU formulation was recently proposed. In this study, we sought to validate the recently proposed Composite SHU as a medium that supports the growth of several bacterial species that are known to grow in normal human urine and/or artificial urine. Comparative growth assays of gram-negative and gram-positive bacteria E. coli, Pseudomonas aeruginosa, Proteus mirabilis, Streptococcus agalactiae, Staphylococcus saprophyticus and Enterococcus faecalis were undertaken using viable bacterial count and optical density measurements over a 48h culture period. Three different SHU formulations were tested in various culture vessels, shaking conditions and volumes and showed that Composite SHU can support the robust growth of gram-negative bacteria but requires supplementation with 0.2% yeast extract to support the growth of gram-positive bacteria. Experiments are also presented that show an unexpected but major influence of P. mirabilis towards the ability to measure bacterial growth in generally accepted multiwell assays using absorbance readings, predicted to have a basis in the release of volatile organic compound(s) from P. mirabilis during growth in Composite SHU medium. This study represents an essential methodological validation of a more chemically defined type of synthetic urine that can be applied to study mechanisms of bacteriuria and we conclude will offer a useful in vitro model to investigate the

  15. Determination of chromatographic dissociation constants of some carbapenem group antibiotics and quantification of these compounds in human urine.

    Science.gov (United States)

    Çubuk Demiralay, Ebru; Koç, Duygu; Daldal, Y Doğan; Alsancak, Güleren; Ozkan, Sibel A

    2014-05-01

    The dissociation constant values (s (s) pKa ) of some carbapenem group drugs (ertapenem, meropenem, doripenem) in different percentages of methanol-water binary mixtures (18, 20 and 22%, v/v) were determined from the mobile phase pH dependence of their retention factor. Evaluation of these data was performed using the NLREG program. From calculated pKa values, the aqueous pKa values of these subtances were calculated by different approaches. Moreover, the correlation established between retention factor and the pH of the water-methanol mobile phase was used to determine the optimum separation conditions. In order to validate the optimized conditions, these drugs were studied in human urine. The chromatographic separation was realized using a Gemini NX C18 column (250 × 4.6 mm i.d., 5 µm particles) and UV detector set at 220 and 295 nm.

  16. Isolation of S-[2-carboxy-1-(1H-imidazol-4-yl)ethyl]cysteine from human urine.

    Science.gov (United States)

    Kinuta, M; Ubuka, T; Yao, K; Futani, S; Fujiwara, M; Kurozumi, Y

    1992-01-01

    S-[2-Carboxy-1-(1H-imidazol-4-yl)ethyl]cysteine (I), a proposed precursor of 3-[(carboxymethyl)thio]-3-(1H-imidazol-4-yl)propanoic acid [Kinuta, Yao, Masuoka, Ohta, Teraoka & Ubuka (1991) Biochem. J. 275, 617-721], was isolated from healthy human urine by using ion-exchange column chromatography. Identification of the isolated compound with compound (I) was performed by physicochemical analyses involving i.r., m.s. and n.m.r. spectrometries as well as high-voltage paper electrophoresis, t.l.c. and paper chromatography. Compound (I) was synthesized in 80% yield by incubation of a reaction mixture containing trans-urocanic acid and 3-fold excess of cysteine at 70-75 degrees C. From these results we suggest that natural thiol compounds such as cysteine and GSH participate in the metabolism of urocanic acid, a key metabolite of L-histidine. PMID:1567378

  17. Cathodic adsorptive stripping voltammetry of drotaverine hydrochloride and its determination in tablets and human urine by differential pulse voltammetry.

    Science.gov (United States)

    Zayed, S I M; Issa, Y M

    2009-04-01

    The stripping voltammetric behaviour of drotaverine hydrochloride (DvCl) was studied using a hanging mercury drop electrode (HMDE). The adsorptive stripping response has been evaluated with respect to pH, accumulation time, accumulation potential, scan rate and other variables. Differential pulse DP mode; over the potential range -400 to -1200 mV, is used in the presence of 0.04 M Britton-Robinson buffer pH 2. Cyclic voltammetric study indicates that the reduction process is irreversible and controlled by adsorption. The response of DP technique is linear over the concentration range 21.70-257.34 ng/ml. Limit of detection and limit of quantification were 3.15 and 10.50 ng/ml, respectively. The proposed method was successfully applied for the determination of the drug in commercial tablets and spiked human urine samples.

  18. An Adaptive Single-Well Stochastic Resonance Algorithm Applied to Trace Analysis of Clenbuterol in Human Urine

    Directory of Open Access Journals (Sweden)

    Shaofei Xie

    2012-02-01

    Full Text Available Based on the theory of stochastic resonance, an adaptive single-well stochastic resonance (ASSR coupled with genetic algorithm was developed to enhance the signal-to-noise ratio of weak chromatographic signals. In conventional stochastic resonance algorithm, there are two or more parameters needed to be optimized and the proper parameters values were obtained by a universal searching within a given range. In the developed ASSR, the optimization of system parameter was simplified and automatic implemented. The ASSR was applied to the trace analysis of clenbuterol in human urine and it helped to significantly improve the limit of detection and limit of quantification of clenbuterol. Good linearity, precision and accuracy of the proposed method ensure that it could be an effective tool for trace analysis and the improvement of detective sensibility of current detectors.

  19. Multiplex analysis inflammatory cytokines in human blood, breath condensate, and urine matrices

    Science.gov (United States)

    Scientific evidence suggests that inflammation is associated with human health effects and health endpoints, yet most studies have focused on human populations that are already considered “unhealthy”.  As such, it is pertinent to measure inflammatory biomarkers in human biologica...

  20. Multiplex analysis inflammatory cytokines in human blood, breath condensate, and urine matrices

    Science.gov (United States)

    Scientific evidence suggests that inflammation is associated with human health effects and health endpoints, yet most studies have focused on human populations that are already considered “unhealthy”.  As such, it is pertinent to measure inflammatory biomarkers in human biologica...

  1. Black Urine

    Directory of Open Access Journals (Sweden)

    Rahim Vakili

    2016-06-01

    Full Text Available A 2-year-old boy was born at term of healthy, non-consanguineous Iranian parents. His mother attended in the clinic with the history of sometimes discoloration of diapers after passing urine. She noticed that first at the age of one month with intensified in recent months. His Physical examination and growth parameters were normal. His mother denied taking any medication (sorbitol, nitrofurantoin, metronidazole, methocarbamol, sena and methyldopa (5. Qualitative urine examination showed dark black discoloration. By this history, alkaptonuria was the most clinical suspicious. A 24-hour-urine sample was collected and sent for quantitative measurements. The urine sample was highly positive for homogentisic acid and negative for porphyrin metabolites.

  2. Spectrophotometric determination of quetiapine fumarate in pharmaceuticals and human urine by two charge-transfer complexation reactions

    Directory of Open Access Journals (Sweden)

    Vinay K.B.

    2012-01-01

    Full Text Available Two simple, rapid and accurate spectrophotometric procedures are proposed for the determination of quetiapine fumarate (QTF in pharmaceuticals and in spiked human urine. The methods are based on charge transfer complexation reactions of free base form of the drug (quetiapine, QTP, as n-electron donor (D, with either p-chloranilic acid (p-CAA (method A or 2,3-dichloro-5,6-dicyanoquinone (DDQ (method B as π-acceptors (A. The coloured charge transfer complexes produced exhibit absorption maxima at 520 and 540 nm, in method A and method B, respectively. The experimental conditions such as reagent concentration, reaction solvent and time have been carefully optimized to achieve the maximum sensitivity. Beer’s law is obeyed over the concentration ranges of 8.0 - 160 and 4.0 - 80.0 μg ml-1, for method A and method B, respectively. The calculated molar absorptivity values are 1.77 × 103 and 4.59 × 103 l mol-1cm-1, respectively, for method A and method B. The Sandell sensitivity values, limits of detection (LOD and quantification (LOQ have also been reported. The stoichiometry of the reaction in both cases was accomplished adopting the limiting logarithmic method and was found to be 1: 2 (D: A. The accuracy and precision of the methods were evaluated on intra-day and inter-day basis. The proposed methods were successfully applied for the determination of QTF in pharmaceutical formulations and spiked human urine.

  3. Analysis of Piroxicam in Pharmaceutical Formulation and Human Urine by Dispersive Liquid–Liquid Microextraction Combined with Spectrophotometry

    Directory of Open Access Journals (Sweden)

    Nakisa Seyyedeh Tutunchi

    2013-02-01

    Full Text Available Purpose: Piroxicam, is non–steroidal anti–inflammatory and analgesic agent, which is widely used in the treatment of patients with rheumatologic disorders. A new analytical approach based on the dispersive liquid–liquid microextraction (DLLME has been developed for the extraction and determination of PX in pharmaceutical preparation and human urine. Methods: From the PX standard solution or solutions prepared from real samples, aliquot volumes were pipetted into centrifuge tubes and mixed with acetate buffer at pH 3.0 and NaCl solution. The contents were subjected to the DLLME, so 700 μL of methanol containing 70 μL of chloroform was injected rapidly into a sample solution. A cloudy solution was rapidly produced and the PX extracted into dispersed fine droplets. The mixture was centrifuged, thus these fine droplets of chloroform were settled. The supernatant aqueous phase was readily decanted, then the remained organic phase was diluted with ethanol and the absorbance measured at 355 ± 3 nm against a reagent blank. Results: The main factors affecting the extraction efficiency such as pH, extraction and disperser solvent types and etc. were studied and optimized systematically. Under optimized conditions, the calibration graphs were linear over the range of 0.2 to 4.8 μg/mL. The limit of detection and relative standard deviation were found to be 0.058 μg/mL and 2.83%, respectively. Relative recoveries in the spiked samples ranged from 97 to 110%. Conclusion: Using the developed method PX can be analyzed in pharmaceutical formulation and human urine sample in a simpler, cheaper and more rapid manner.

  4. Analysis of Piroxicam in Pharmaceutical Formulation and Human Urine by Dispersive Liquid–Liquid Microextraction Combined with Spectrophotometry

    Science.gov (United States)

    Bavili Tabrizi, Ahad; Seyyedeh Tutunchi, Nakisa

    2013-01-01

    Purpose: Piroxicam, is non–steroidal anti–inflammatory and analgesic agent, which is widely used in the treatment of patients with rheumatologic disorders. A new analytical approach based on the dispersive liquid–liquid microextraction (DLLME) has been developed for the extraction and determination of PX in pharmaceutical preparation and human urine. Methods: From the PX standard solution or solutions prepared from real samples, aliquot volumes were pipetted into centrifuge tubes and mixed with acetate buffer at pH 3.0 and NaCl solution. The contents were subjected to the DLLME, so 700 µL of methanol containing 70 µL of chloroform was injected rapidly into a sample solution. A cloudy solution was rapidly produced and the PX extracted into dispersed fine droplets. The mixture was centrifuged, thus these fine droplets of chloroform were settled. The supernatant aqueous phase was readily decanted, then the remained organic phase was diluted with ethanol and the absorbance measured at 355 ± 3 nm against a reagent blank. Results: The main factors affecting the extraction efficiency such as pH, extraction and disperser solvent types and etc. were studied and optimized systematically. Under optimized conditions, the calibration graphs were linear over the range of 0.2 to 4.8 µg/mL. The limit of detection and relative standard deviation were found to be 0.058 µg/mL and 2.83%, respectively. Relative recoveries in the spiked samples ranged from 97 to 110%. Conclusion: Using the developed method PX can be analyzed in pharmaceutical formulation and human urine sample in a simpler, cheaper and more rapid manner. PMID:24312810

  5. Spectrophotometric determination of certain CNS stimulants in dosage forms and spiked human urine via derivatization with 2,4-Dinitrofluorobenzene

    Directory of Open Access Journals (Sweden)

    Saad Samar

    2011-10-01

    Full Text Available Abstract A new spectrophotometric method is developed for the determination of phenylpropanolamine HCl (PPA, ephedrine HCl (EPH and pseudoephedrine HCl (PSE in pharmaceutical preparations and spiked human urine. The method involved heat-catalyzed derivatization of the three drugs with 2,4-dinitrofluorobenzene (DNFB producing a yellow colored product peaking at 370 nm for PPA and 380 nm for EPH and PSE, respectively. The absorbance concentration plots were rectilinear over the range of 2-20 for PPA and 1-14 μg/mL for both of EPH and PSE, respectively. The limit of detection (LOD values were 0.20, 0.13 and 0.20 μg/mL for PPA, EPH and PSE, respectively and limit of quantitation (LOQ values of 0.60 and 0.40 and 0.59 μg/mL for PPA, EPH and PSE, respectively. The analytical performance of the method was fully validated and the results were satisfactory. The proposed method was successfully applied to the determination of the three studied drugs in their commercial dosage forms including tablets, capsules and ampoules with good percentage recoveries. The proposed method was further applied for the determination of PSE in spiked human urine with a mean percentage recovery of 108.17 ± 1.60 for (n = 3. Statistical comparison of the results obtained with those of the comparison methods showed good agreement and proved that there was no significant difference in the accuracy and precision between the two methods. The mechanism of the reaction pathway was postulated.

  6. Flow injection on-line dilution for multi-element determination in human urine with detection by inductively coupled plasma mass spectrometry

    DEFF Research Database (Denmark)

    Wang, J.H.; Hansen, E.H.; Gammelgaard, Bente

    2001-01-01

    A simple flow injection on-line dilution procedure with detection by inductively coupled plasma mass spectrometry (ICP-MS) was developed for the determination of copper, zinc, arsenic, lead, selenium, nickel and molybdenum in human urine. Matrix effects were minimized by employing a dilution fact...... of 16.5 with on-line standard addition, and Rh-103 was used as internal standard to compensate for signal fluctuation. The procedure was validated by the analysis of two standard reference materials SRM 2670 (NIST) and Seronorm (TM) Trace Elements in Urine. Recovery experiments were pet...... human urine samples. No correlations between the concentrations of the elements were observed. (C) 2001 Elsevier Science B.V. All rights reserved...

  7. Cryptococcus neoformans en excretas de palomas del perímetro urbano de Cali.

    Directory of Open Access Journals (Sweden)

    Luz Dary Caicedo B

    2009-10-01

    Full Text Available En el perímetro urbano de Santiago de Cali, Colombia, se realizó un estudio con excretas de palomas, con el fin de establecer la presencia de Cryptococcus neoformans; para aislar el hongo se utilizó agar semilla de girasol. Se analizó si la presencia del hongo se relacionaba con factores como la cantidad de excretas, el tipo de nido, el número de palomas y el pH. De 119 muestras, 59 (49.6% presentaron la levadura. Todos los aislamientos correspondieron a C. neoformans var neoformans. Se estableció que la levadura tiene una amplia distribución en la ciudad y que hay mayor probabilidad de encontrarla en excretas acumuladas (p

  8. Thorium and uranium contents in human urine: influence of age and residential area.

    Science.gov (United States)

    Al-Jundi, J; Werner, E; Roth, P; Höllriegl, V; Wendler, I; Schramel, P

    2004-01-01

    Inductively coupled plasma mass spectrometry (ICP-MS) has been used for the determination of (232)Th and (238)U in urine of unexposed Jordanian subjects living in six cities. The range of (232)Th excretion in all subjects was found to be 1.4-640 microBq d(-1) with an average of 34.8 microBq d(-1) (geometric mean 15.8 microBq d(-1)). Results showed no statistically significant correlation with age and residential area. The average value obtained is in agreement with levels considered normal in some recent publications. The average value of (238)U in all samples was found to be 3955 microBq d(-1) (geometric mean 1107 microBq d(-1)), which is higher than reported figures from Germany and India, but in agreement with those figures given in ICRP publication, number 23. The mean values of the different groups were found to be proportional to age up to 60 years. A noticeable drop is observed for subjects greater than 60 years old.

  9. Diurnal rhythms in the human urine metabolome during sleep and total sleep deprivation.

    Science.gov (United States)

    Giskeødegård, Guro F; Davies, Sarah K; Revell, Victoria L; Keun, Hector; Skene, Debra J

    2015-10-09

    Understanding how metabolite levels change over the 24 hour day is of crucial importance for clinical and epidemiological studies. Additionally, the association between sleep deprivation and metabolic disorders such as diabetes and obesity requires investigation into the links between sleep and metabolism. Here, we characterise time-of-day variation and the effects of sleep deprivation on urinary metabolite profiles. Healthy male participants (n = 15) completed an in-laboratory study comprising one 24 h sleep/wake cycle prior to 24 h of continual wakefulness under highly controlled environmental conditions. Urine samples were collected over set 2-8 h intervals and analysed by (1)H NMR spectroscopy. Significant changes were observed with respect to both time of day and sleep deprivation. Of 32 identified metabolites, 7 (22%) exhibited cosine rhythmicity over at least one 24 h period; 5 exhibiting a cosine rhythm on both days. Eight metabolites significantly increased during sleep deprivation compared with sleep (taurine, formate, citrate, 3-indoxyl sulfate, carnitine, 3-hydroxyisobutyrate, TMAO and acetate) and 8 significantly decreased (dimethylamine, 4-DTA, creatinine, ascorbate, 2-hydroxyisobutyrate, allantoin, 4-DEA, 4-hydroxyphenylacetate). These data indicate that sampling time, the presence or absence of sleep and the response to sleep deprivation are highly relevant when identifying biomarkers in urinary metabolic profiling studies.

  10. Simultaneous GC-ECNICI-MS measurement of nitrite, nitrate and creatinine in human urine and plasma in clinical settings.

    Science.gov (United States)

    Hanff, Erik; Lützow, Moritz; Kayacelebi, Arslan Arinc; Finkel, Armin; Maassen, Mirja; Yanchev, Georgi Radoslavov; Haghikia, Arash; Bavendiek, Udo; Buck, Anna; Lücke, Thomas; Maassen, Norbert; Tsikas, Dimitrios

    2017-03-15

    Creatinine in urine is a useful biochemical parameter to correct the urinary excretion rate of endogenous and exogenous substances. Nitrite (ONO(-)) and nitrate (ONO2(-)) are metabolites of nitric oxide (NO), a signalling molecule with multiple biological functions. Under certain and standardized conditions, the concentration of nitrate in the urine is a suitable measure of whole body NO synthesis. The urinary nitrate-to-nitrite molar ratio (UNOxR) may indicate nitrite-dependent renal carbonic anhydrase (CA) activity. In clinical studies, urine is commonly collected by spontaneous micturition. In those cases the nitrate and nitrite excretion must be corrected for creatinine excretion. Pentafluorobenzyl (PFB) bromide (PFB-Br) is a useful derivatization reagent of numerous inorganic and organic compounds, including urinary nitrite, nitrate and creatinine, for highly sensitive and specific quantitation by GC-MS. Here, we report on the simultaneous PFB-Br derivatization (60min, 50°C) of ONO(-), O(15)NO(-), ONO2(-), O(15)NO2(-), creatinine (do-Crea) and [methylo-(2)H3]creatinine (d3-Crea) in acetonic dilutions of native human urine and plasma samples (4:1, v/v) and their simultaneous quantification by GC-MS as PFBNO2, PFB(15)NO2, PFBONO2, PFBO(15)NO2, do-Crea-PFB and d3-Crea-PFB, respectively. Electron capture negative-ion chemical ionization (ECNICI) of these derivatives generates anions due to [M-PFB](-), i.e., the starting analytes. Quantification is performed by selected-ion monitoring (SIM) of m/z 46 (ONO(-)), m/z 47 (O(15)NO(-)), m/z 62 (ONO2(-)), m/z 63 (O(15)NO2(-)), m/z 112 (do-Crea), and m/z 115 (d3-Crea). Retention times were 2.97min for PFB-ONO2/PFB-O(15)NO2, 3.1min for PFB-NO2/PFB-(15)NO2, and 6.7min for do-Crea-PFB/d3-Crea-PFB. We used this method to investigate the effects of long-term oral NaNO3 or NaCl (serving as placebo) supplementation (each 0.1mmol/kg body weight per day for 3 weeks) on creatinine excretion and UNOxR in 17 healthy young men

  11. Comparative Study of Seven Commercial Kits for Human DNA Extraction from Urine Samples Suitable for DNA Biomarker-Based Public Health Studies

    Science.gov (United States)

    El Bali, Latifa; Diman, Aurélie; Bernard, Alfred; Roosens, Nancy H. C.; De Keersmaecker, Sigrid C. J.

    2014-01-01

    Human genomic DNA extracted from urine could be an interesting tool for large-scale public health studies involving characterization of genetic variations or DNA biomarkers as a result of the simple and noninvasive collection method. These studies, involving many samples, require a rapid, easy, and standardized extraction protocol. Moreover, for practicability, there is a necessity to collect urine at a moment different from the first void and to store it appropriately until analysis. The present study compared seven commercial kits to select the most appropriate urinary human DNA extraction procedure for epidemiological studies. DNA yield has been determined using different quantification methods: two classical, i.e., NanoDrop and PicoGreen, and two species-specific real-time quantitative (q)PCR assays, as DNA extracted from urine contains, besides human, microbial DNA also, which largely contributes to the total DNA yield. In addition, the kits giving a good yield were also tested for the presence of PCR inhibitors. Further comparisons were performed regarding the sampling time and the storage conditions. Finally, as a proof-of-concept, an important gene related to smoking has been genotyped using the developed tools. We could select one well-performing kit for the human DNA extraction from urine suitable for molecular diagnostic real-time qPCR-based assays targeting genetic variations, applicable to large-scale studies. In addition, successful genotyping was possible using DNA extracted from urine stored at −20°C for several months, and an acceptable yield could also be obtained from urine collected at different moments during the day, which is particularly important for public health studies. PMID:25365790

  12. Comparative study of seven commercial kits for human DNA extraction from urine samples suitable for DNA biomarker-based public health studies.

    Science.gov (United States)

    El Bali, Latifa; Diman, Aurélie; Bernard, Alfred; Roosens, Nancy H C; De Keersmaecker, Sigrid C J

    2014-12-01

    Human genomic DNA extracted from urine could be an interesting tool for large-scale public health studies involving characterization of genetic variations or DNA biomarkers as a result of the simple and noninvasive collection method. These studies, involving many samples, require a rapid, easy, and standardized extraction protocol. Moreover, for practicability, there is a necessity to collect urine at a moment different from the first void and to store it appropriately until analysis. The present study compared seven commercial kits to select the most appropriate urinary human DNA extraction procedure for epidemiological studies. DNA yield has been determined using different quantification methods: two classical, i.e., NanoDrop and PicoGreen, and two species-specific real-time quantitative (q)PCR assays, as DNA extracted from urine contains, besides human, microbial DNA also, which largely contributes to the total DNA yield. In addition, the kits giving a good yield were also tested for the presence of PCR inhibitors. Further comparisons were performed regarding the sampling time and the storage conditions. Finally, as a proof-of-concept, an important gene related to smoking has been genotyped using the developed tools. We could select one well-performing kit for the human DNA extraction from urine suitable for molecular diagnostic real-time qPCR-based assays targeting genetic variations, applicable to large-scale studies. In addition, successful genotyping was possible using DNA extracted from urine stored at -20°C for several months, and an acceptable yield could also be obtained from urine collected at different moments during the day, which is particularly important for public health studies.

  13. 2,2-bis(4-Chlorophenyl)acetic acid (DDA), a water-soluble urine biomarker of DDT metabolism in humans.

    Science.gov (United States)

    Chen, Zhenshan; Maartens, Francois; Vega, Helen; Kunene, Simon; Gumede, Jonathan; Krieger, Robert I

    2009-01-01

    DDT metabolism in humans yields DDA as the principal urinary metabolite and potential exposure biomarker. A method for DDA analysis in human urine was developed using pentafluorobenzyl bromide and diisopropylethyl amine. Dried hexane extracts were reacted for 1 hour at room temperature. The stable DDA-pentafluorobenzyl-ester derivative was analyzed by gas chromatography-electron capture detector (GC-ECD) and confirmed by gas chromatography-mass spectrometry (GC-MS) in selective ion monitoring mode. The limit of detection for DDA was 0.1 microg/L urine by GC-ECD and 2 microg/L urine by GC-MS, with a relative standard deviation of 12%. Urine specimens from DDT applicators in Swaziland and South Africa were analyzed to evaluate the method. The mean DDA levels during the spray season and post season were 59 and 11 microg/L, respectively. These results must be interpreted cautiously because different groups of workers provided urine specimens in each case. The DDA urinalysis may be a feasible monitoring strategy for low-level occupational and residential DDT exposure assessment in antimalaria campaigns.

  14. Determination of terbuthylazine and desethylterbuthylazine in human urine and hair samples by eletrospray ionization-liquid chromatography/triple quadrupole mass spectrometry.

    Science.gov (United States)

    Mercadante, Rosa; Polledri, Elisa; Fustinoni, Silvia

    2012-08-01

    Terbuthylazine (TBA) is a widely applied herbicide and an environmental contaminant. Following its use, humans, such as agricultural workers and rural residents, may be exposed. An isotope-dilution liquid chromatography coupled to electrospray-tandem mass spectrometry method for the determination of TBA, and its metabolite desethylterbuthylazine (DET) in human urine and hair was developed and validated. Under the optimised conditions, analytes were extracted from urine using a solid phase cartridge or from hair by sonication in methanol. Analytes were separated using a C18 reversed-phase chromatographic column and quantified, after positive ionization using a heated electrospray source, by a triple quadrupole mass detector in the selected reaction monitoring mode. Validation showed linear dynamic ranges up to 100 μg/L or 5.00 ng/mg hair, inter- and intra-run precisions <7%, and accuracies within 12% of spiked concentrations. Limits of quantification were 0.25 μg/L in urine and 0.01 ng/mg hair for both TBA and DET. Matrix effect evaluation showed that the isotope dilution approach allowed for the control of bias sources. TBA and DET were determined in specimens of agriculture workers exposed to TBA using the validated method. Hair samples contained TBA levels in the low nanogram per milligram range, and urine samples contained DET levels in the low microgram per liter range. Conversely, TBA levels in urine samples and DET levels in hair samples were always below the limit of quantification.

  15. Ketones urine test

    Science.gov (United States)

    Ketone bodies - urine; Urine ketones; Ketoacidosis - urine ketones test; Diabetic ketoacidosis - urine ketones test ... Urine ketones are usually measured as a "spot test." This is available in a test kit that ...

  16. Simultaneous determination of six mercapturic acid metabolites of volatile organic compounds in human urine.

    Science.gov (United States)

    Ding, Yan S; Blount, Benjamin C; Valentin-Blasini, Liza; Applewhite, Heather S; Xia, Yang; Watson, Clifford H; Ashley, David L

    2009-06-01

    The widespread exposure to potentially harmful volatile organic compounds (VOCs) merits the development of practical and accurate exposure assessment methods. Measuring the urinary concentrations of VOC mercapturic acid (MA) metabolites provides noninvasive and selective information about recent exposure to certain VOCs. We developed a liquid chromatography-tandem mass spectrometry method for quantifying urinary levels of six MAs: N-acetyl-S-(2-carboxyethyl)-L-cysteine (CEMA), N-acetyl-S-(3-hydroxypropyl)-L-cysteine (HPMA), N-acetyl-S-(2-hydroxy-3-butenyl)-L-cysteine (MHBMA), N-acetyl-S-(3,4-dihydroxybutyl)-L-cysteine (DHBMA), N-acetyl-S-(2-hydroxyethyl)-L-cysteine (HEMA), and N-acetyl-S-(phenyl)-L-cysteine (PMA). The method provides good accuracy (102% mean accuracy) and high precision (3.5% mean precision). The sensitivity (limits of detection of 0.01-0.20 microg/L) and wide dynamic detection range (0.025-500 microg/L) make this method suitable for assessing VOC exposure of minimally exposed populations and those with significant exposures, such as cigarette smokers. We used this method to quantify MA levels in urine collected from smokers and nonsmokers. Median levels of creatinine-corrected CEMA, HPMA, MHBMA, DHBMA, HEMA, and PMA among nonsmokers (n = 59) were 38.1, 24.3, 21.3, 104.7, 0.9, and 0.5 microg/g creatinine, respectively. Among smokers (n = 61), median levels of CEMA, HPMA, MHBMA, DHBMA, HEMA, and PMA were 214.4, 839.7, 10.2, 509.7, 2.2, and 0.9 microg/g creatinine, respectively. All VOC MAs measured were higher among smokers than among nonsmokers, with the exception of MHBMA.

  17. Isolation and Identification of Exosomes in Human Urine%人尿液中exosomes的分离及鉴定

    Institute of Scientific and Technical Information of China (English)

    李艳艳; 张渊; 代义; 邱峰; 秦霞; 邱宗荫

    2011-01-01

    Objective To isolate exosomes from human urine and identify their ultrastructural morphology and immune characteristics.Methods Exosomes were isolated from the mixed urine of healthy volunteers by ultracentrifugation and identified by electron microscopy with negative staining and immunoelectron microscopy, from which protein component was separated by 1-D SDS-PAGE.Results The exosmoes with high purity was obtained by further centrifugation of the supematant of urine, collected after centrifugation at 4℃, 17 000 × g for 15 min, at 4℃, 200 000 × g for 1 h.Negative staining under electron microscope showed that the exosomes were flat or spherical corpuscula, in a diameter of about 60 nm.Colloidal gold particles were distributed evenly on the surface of exosomes, which were shown as clear black spots due to high electron density under immunoelectron microscope.1-D SDSPAGE showed a majority of exosome bands with high abundance were 90 000 ~ 110 000, several clear bands with relative molecular masses of 30 000 ~ 60 000, and few bands with relative molecular masses of less than 30 000.Conclusion A method for isolation of exosomes from human urine was developed, which laid a foundation of further study on potential biomarkers of diseases.%目的 分离人尿液中的exosomes,并对其超微形态学和免疫特性进行鉴定.方法 以超速离心法分离健康志愿者混合尿液中的exosomes,通过负染电镜和免疫电镜技术对exosomes进行鉴定,运用1-D SDS-PAGE对其蛋白质组分进行分离.结果 4℃,17000×g离心15min后的上清液于4℃,200000×g离心1h可得到纯度较高的exosomes.负染电镜可见exosome,为扁平或球形小体,直径主要在60nm左右;免疫电镜显示,黑色胶体金颗粒均匀分布在exosomes球形体表面,因其具有很高的电子密度而呈现为清晰可辨的黑点;1-D SDS-PAGE分析显示,exosomes高丰度条带主要分布于相对分子质量90000-110000)之间,且出现了数条清

  18. Human Urine Derived Stem Cells in Combination with β-TCP Can Be Applied for Bone Regeneration.

    Directory of Open Access Journals (Sweden)

    Junjie Guan

    Full Text Available Bone tissue engineering requires highly proliferative stem cells that are easy to isolate. Human urine stem cells (USCs are abundant and can be easily harvested without using an invasive procedure. In addition, in our previous studies, USCs have been proved to be able to differentiate into osteoblasts, chondrocytes, and adipocytes. Therefore, USCs may have great potential and advantages to be applied as a cell source for tissue engineering. However, there are no published studies that describe the interactions between USCs and biomaterials and applications of USCs for bone tissue engineering. Therefore, the objective of the present study was to evaluate the interactions between USCs with a typical bone tissue engineering scaffold, beta-Tricalcium Phosphate (β-TCP, and to determine whether the USCs seeded onto β-TCP scaffold can promote bone regeneration in a segmental femoral defect of rats. Primary USCs were isolated from urine and seeded on β-TCP scaffolds. Results showed that USCs remained viable and proliferated within β-TCP. The osteogenic differentiation of USCs within the scaffolds was demonstrated by increased alkaline phosphatase activity and calcium content. Furthermore, β-TCP with adherent USCs (USCs/β-TCP were implanted in a 6-mm critical size femoral defect of rats for 12 weeks. Bone regeneration was determined using X-ray, micro-CT, and histologic analyses. Results further demonstrated that USCs in the scaffolds could enhance new bone formation, which spanned bone defects in 5 out of 11 rats while β-TCP scaffold alone induced modest bone formation. The current study indicated that the USCs can be used as a cell source for bone tissue engineering as they are compatible with bone tissue engineering scaffolds and can stimulate the regeneration of bone in a critical size bone defect.

  19. Human Urine Derived Stem Cells in Combination with β-TCP Can Be Applied for Bone Regeneration.

    Science.gov (United States)

    Guan, Junjie; Zhang, Jieyuan; Li, Haiyan; Zhu, Zhenzhong; Guo, Shangchun; Niu, Xin; Wang, Yang; Zhang, Changqing

    2015-01-01

    Bone tissue engineering requires highly proliferative stem cells that are easy to isolate. Human urine stem cells (USCs) are abundant and can be easily harvested without using an invasive procedure. In addition, in our previous studies, USCs have been proved to be able to differentiate into osteoblasts, chondrocytes, and adipocytes. Therefore, USCs may have great potential and advantages to be applied as a cell source for tissue engineering. However, there are no published studies that describe the interactions between USCs and biomaterials and applications of USCs for bone tissue engineering. Therefore, the objective of the present study was to evaluate the interactions between USCs with a typical bone tissue engineering scaffold, beta-Tricalcium Phosphate (β-TCP), and to determine whether the USCs seeded onto β-TCP scaffold can promote bone regeneration in a segmental femoral defect of rats. Primary USCs were isolated from urine and seeded on β-TCP scaffolds. Results showed that USCs remained viable and proliferated within β-TCP. The osteogenic differentiation of USCs within the scaffolds was demonstrated by increased alkaline phosphatase activity and calcium content. Furthermore, β-TCP with adherent USCs (USCs/β-TCP) were implanted in a 6-mm critical size femoral defect of rats for 12 weeks. Bone regeneration was determined using X-ray, micro-CT, and histologic analyses. Results further demonstrated that USCs in the scaffolds could enhance new bone formation, which spanned bone defects in 5 out of 11 rats while β-TCP scaffold alone induced modest bone formation. The current study indicated that the USCs can be used as a cell source for bone tissue engineering as they are compatible with bone tissue engineering scaffolds and can stimulate the regeneration of bone in a critical size bone defect.

  20. Liquid-Based Urine Cytology as a Tool for Detection of Human Papillomavirus, Mycoplasma spp., and Ureaplasma spp. in Men

    Science.gov (United States)

    Kawaguchi, Shohei; Shigehara, Kazuyoshi; Shimamura, Masayoshi; Nakashima, Takao; Sugimoto, Kazuhiro; Nakashima, Kazufumi; Furubayashi, Keiichi; Namiki, Mikio

    2012-01-01

    Liquid-based urine cytology (LB-URC) was evaluated for cytological diagnosis and detection of human papillomavirus (HPV), Mycoplasma, and Ureaplasma. Midstream urine samples were collected from 141 male patients with urethritis and 154 controls without urethritis, and sediment cells were preserved in liquid-based cytology solution. Urethral swabs from urethritis patients were tested for the presence of Neisseria gonorrhoeae and Chlamydia trachomatis. Papanicolaou tests were performed for cytological evaluation. HPV, Mycoplasma, and Ureaplasma genomes were determined by PCR-based methods, and localization of HPV DNA in urothelial cells was examined by in situ hybridization (ISH). The β-globin gene was positive in 97.9% of LB-URC samples from urethritis patients and in 97.4% of control samples, suggesting that high-quality cellular DNA was obtained from the LB-URC samples. HPV DNA was detected in 29 (21.0%) urethritis cases and in five (3.3%) controls (P Cytological evaluations could be performed for 92.1% of urethritis patients and 64.3% of controls. Morphological changes suggestive of HPV infection were seen in 20.7% of the HPV-positive samples, and ISH demonstrated the presence of HPV DNA in both squamous and urothelial cells in HPV-positive samples. Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma parvum, and Ureaplasma urealyticum were detected in 14.5%, 10.9%, 6.5%, and 12.3% of urethritis patients, respectively. The prevalence rates of these microorganisms (except Ureaplasma parvum) were significantly higher in urethritis cases than controls (P < 0.05). LB-URC is applicable for detection of HPV, Mycoplasma, and Ureaplasma. HPV infection occurs in urothelial cells, especially in gonococcal urethritis. PMID:22135257

  1. Relevance of the detection of Streptococcus pneumoniae antigen in human urine in the diagnosis of lower respiratory tract infections

    National Research Council Canada - National Science Library

    Sorlózano, Antonio; Cedeño, Sindy; Gutiérrez-Fernández, José; Polo, Purificación; Navarro, José María

    2013-01-01

    Techniques membrane antigen immunochromatographic detecting in urine the pneumococcal polysaccharide C, have developed significantly, increasing requests for antigenuria to clinical microbiology laboratories...

  2. Influence of human urine to antimicrobial susceptibility of clinical isolates of Klebsiella pneumoniae and Escherichia coli producing β-lactamase of different types

    Directory of Open Access Journals (Sweden)

    Ž. Žagar

    2007-02-01

    Full Text Available The purpose of the study was to determine the influence of human urine on the antibiotic susceptibilities of Klebsiella pneumoniae and Escherichia coli strains producing different types of extended-spectrum β-lactamases (ESBL. The study was performed on 26 ESBL negative strains of K. pneumoniae, 80 K. pneumoniae strains producing SHV-ESBLs (52-SHV-5, 31- SHV-2 and 7- SHV-12, 94 E. coli strains harbouring TEM- ESBLs and 14 E. coli strains possessing CTX-M group 1 β-lactamases. The minimum inhibitory concentrations of amoxycillin alone and combined with clavulanate (co-amoxilcav, cephalexin, cefuroxime, ceftazidime, cefotaxime, ceftriaxone, cefepime, gentamicin and ciprofloxacin were performed in parallel in Mueller-Hinton broth and urine by broth microdilution method. With ESBL negative strains, urine increased MIC90 of amoxycillin alone and combined with clavulanate, cephalexin, cefuroxime, ceftazidime, cefotaxime, ceftriaxone, cefepime, gentamicin and ciprofloxacin. Against SHV-5 producers, an increase in MIC90 was observed with cefotaxime, cefepime and ciprofloxacin when the test was performed in urine. SHV-2 producers showed elevated MIC90 of ceftazidime, cefotaxime, ceftriaxone and cefepime in the presence of urine, in contrast to SHV-12 producers which displayed elevated MIC90 only for cefotaxime. Urine increased MIC90 of amoxycillin/clavulanate, ceftazidime and cefepime against CTX-M producers, and of amoxycillin/clavulanate, cefotaxime, ceftriaxone, cefepime and ciprofloxacin for TEM producers. According to our results the activity of antibiotics used for the treatment of urinary tract infection could be overestimated by a standard in vitro testing. However, most of antibiotics used for the treatment of urinary tract infection achieve very high concentration in urine and that could abrogate the reduction of antimicrobial activity by biological fluid.

  3. Identification and quantification of flavonoids in human urine samples by column switching liquid chromatography coupled to atmospheric pressure chemical ionization mass spectrometry

    DEFF Research Database (Denmark)

    Nielsen, S. E.; Freese, R.; Cornett, Claus

    2000-01-01

    by column-switching, using the first column (a Zorbax 300SB C-3 column) for sample cleanup and eluting the heart-cut flavonoid fraction onto the second column (a Zorbax SE C-18 column) for separation and detection by ultraviolet and atmospheric pressure chemical ionization MS using single ion monitoring...... of variation for the analysis of the 12 different flavonoids in quality control urine samples were 12.3% on average (range 11.0-13.7%, n = 24, reproducibility) and the repeatability of the assay were 5.0% (mean, range 0.1-14.8%, it = 12). A subset of 10 urine samples from a human dietary intervention study...

  4. Combination of hollow fiber-based liquid-phase microextraction with sweeping techniques in micellar electrokinetic chromatography for the determination of Strychnos alkaloids in human urine

    Institute of Scientific and Technical Information of China (English)

    Xiao Huan Zang; Cai Rui Li; Qiu Hua Wu; Chun Wang; Dan Dan Han; Zhi Wang

    2007-01-01

    A new method for the enrichment of Strychnos alkaloids in biological samples via liquid-phase rnicroextraction (LPME) based on porous polypropylene hollow fibers in combination with on-line sweeping in micellar electrokinetic chromatography was developed. The calibration curve was linear over the range of 20-200 ng mL-1 for both strychnine and brucine in human urine sample. The detection limits (S/N = 3:1) for strychnine and brucine were 1 ng mL-1 and 2 ng mL-1, respectively. The LPME-sweeping method has been successfully applied to the analysis of strychnine and brucine in real urine samples.

  5. Occurrence of artificial sweeteners in human liver and paired blood and urine samples from adults in Tianjin, China and their implications for human exposure.

    Science.gov (United States)

    Zhang, Tao; Gan, Zhiwei; Gao, Chuanzi; Ma, Ling; Li, Yanxi; Li, Xiao; Sun, Hongwen

    2016-09-14

    In this study, acesulfame (ACE), saccharin (SAC) and cyclamate (CYC) were found in all paired urine and blood samples collected from healthy adults, with mean values of 4070, 918 and 628 ng mL(-1), respectively, in urine and 9.03, 20.4 and 0.72 ng mL(-1), respectively, in blood. SAC (mean: 84.4 ng g(-1)) and CYC (4.29 ng g(-1)) were detectable in all liver samples collected from liver cancer patients, while ACE was less frequently detected. Aspartame (ASP) was not found in any analyzed human sample, which can be explained by the fact that this chemical metabolized rapidly in the human body. Among all adults, significantly positive correlations between SAC and CYC levels were observed (p < 0.001), regardless of human matrices. Nevertheless, no significant correlations between concentrations of SAC (or CYC) and ACE were found in any of the human matrices. Our results suggest that human exposure to SAC and CYC is related, whereas ACE originates from a discrete source. Females (or young adults) were exposed to higher levels of SAC and CYC than males (or elderly). The mean renal clearance of SAC was 730 mL per day per kg in adults, which was significantly (p < 0.001) lower than those for CYC (10 800 mL per day per kg) and ACE (10 300 mL per day per kg). The average total daily intake of SAC and ACE was 9.27 and 33.8 μg per kg bw per day, respectively.

  6. Quantification of 3-MCPD and its mercapturic metabolite in human urine: validation of an LC-MS-MS method and its application in the general population.

    Science.gov (United States)

    Andreoli, Roberta; Cirlini, Martina; Mutti, Antonio

    2015-06-01

    A new method for the simultaneous quantitative determination in human urine of 3-monochloropropane-1,2-diol (3-MCPD), a toxic food contaminant, and its metabolite, 2,3-dihydroxypropyl mercapturic acid (DHPMA), was developed and validated. After urine dilution, the analytes were separated on an Atlantis®dC18 column and quantified by liquid chromatography-tandem mass spectrometry using isotopically labelled internal standards. The limits of quantification (S/N = 10) were 1.90 and 2.21 μg/L for 3-MCPD and DHPMA, respectively. Intra- and inter-day precision were lower than 6 % for each compound. Matrix effects were evaluated. Due to the high sensitivity and good accuracy of the method, 3-MCPD and DHPMA were found in 67 and 100 % of urine samples of healthy subjects, respectively.

  7. Water-compatible molecularly imprinted polymer as a sorbent for the selective extraction and purification of adefovir from human serum and urine.

    Science.gov (United States)

    Pourfarzib, Mojgan; Dinarvand, Rasoul; Akbari-Adergani, Behrouz; Mehramizi, Ali; Rastegar, Hossein; Shekarchi, Maryam

    2015-05-01

    A molecularly imprinted polymer has been synthesized to specifically extract adefovir, an antiviral drug, from serum and urine by dispersive solid-phase extraction before high-performance liquid chromatography with UV analysis. The imprinted polymers were prepared by bulk polymerization by a noncovalent imprinting method that involved the use of adefovir (template molecule) and functional monomer (methacrylic acid) complex prior to polymerization, ethylene glycol dimethacrylate as cross-linker, and chloroform as porogen. Molecular recognition properties, binding capacity, and selectivity of the molecularly imprinted polymers were evaluated and the results show that the obtained polymers have high specific retention and enrichment for adefovir in aqueous medium. The new imprinted polymer was utilized as a molecular sorbent for the separation of adefovir from human serum and urine. The serum and urine extraction of adefovir by the molecularly imprinted polymer followed by high-performance liquid chromatography showed a linear calibration curve in the range of 20-100 μg/L with excellent precisions (2.5 and 2.8% for 50 μg/L), respectively. The limit of detection and limit of quantization were determined in serum (7.62 and 15.1 μg/L), and urine (5.45 and 16 μg/L). The recoveries for serum and urine samples were found to be 88.2-93.5 and 84.3-90.2%, respectively.

  8. Monolithic silica spin column extraction and simultaneous derivatization of amphetamines and 3,4-methylenedioxyamphetamines in human urine for gas chromatographic-mass spectrometric detection.

    Science.gov (United States)

    Nakamoto, Akihiro; Nishida, Manami; Saito, Takeshi; Kishiyama, Izumi; Miyazaki, Shota; Murakami, Katsunori; Nagao, Masataka; Namura, Akira

    2010-02-19

    A simple, sensitive, and specific method with gas chromatography-mass spectrometry was developed for simultaneous extraction and derivatization of amphetamines (APs) and 3,4-methylenedioxyamphetamines (MDAs) in human urine by using a monolithic silica spin column. All the procedures, such as sample loading, washing, and elution were performed by centrifugation. APs and MDAs in urine were adsorbed on the monolithic silica and derivatized with propyl chloroformate in the column. Methamphetamine-d(5) was used as an internal standard. The linear ranges were 0.01-5.0 microg mL(-1) for methamphetamine (MA) and 3,4-methylenedioxymethamphetamine (MDMA) and 0.02-5.0 microg mL(-1) for amphetamine (AP) and 3,4-methylenedioxyamphetamine (MDA) (coefficient of correlation > or = 0.995). The recovery of APs and MDAs in urine was 84-94%, and the relative standard deviation of the intra- and interday reproducibility for urine samples containing 0.1, 1.0, and 4.0 microg mL(-1) of APs and MDAs ranged from 1.4% to 13.6%. The lowest detection limit (signal-to-noise ratio > or = 3) in urine was 5 ng mL(-1) for MA and MDMA and 10 ng mL(-1) for AP and MDA. The proposed method can be used to perform simultaneous extraction and derivatization on spin columns that have been loaded with a small quantity of solvent by using centrifugation.

  9. Metabolite profiling of human urine by CE-ESI-MS using separation electrolytes at low pH.

    Science.gov (United States)

    Benavente, Fernando; van der Heijden, Rob; Tjaden, Ubbo R; van der Greef, Jan; Hankemeier, Thomas

    2006-11-01

    We investigated the potential of CE coupled to electrospray MS (CE-ESI-MS) in metabolite profiling of human urine without any sample prefractionation step. A heterogeneous mixture of biologically relevant compounds covering a broad range of physicochemical properties was used to optimize separation conditions in fused-silica capillaries. A running electrolyte containing 50 mM of acetic acid and 50 mM of formic acid at pH 2.5 was used for the CE separations. A sheath-flow electrospray interface was employed for CE-ESI-MS analysis. Sheath liquids containing 80:20 v/v methanol/water with 0.1% v/v of acetic acid or 60:40 v/v isopropanol/water with 0.5% v/v of ammonia were selected for optimum detection in positive and negative ESI modes, respectively. Reproducibility and sensitivity were studied, and strategies for identification of the separated urinary compounds are suggested. We report major advantages and disadvantages of CE-ESI-MS for metabolite profiling of human body fluids. This work may be regarded as a first step in the use of CE-ESI-MS for reliable differential analysis of body fluids from healthy and diseased individuals.

  10. Paralytic shellfish toxins in clinical matrices: Extension of AOAC official method 2005.06 to human urine and serum and application to a 2007 case study in Maine

    Science.gov (United States)

    DeGrasse, Stacey; Rivera, Victor; Roach, John; White, Kevin; Callahan, John; Couture, Darcie; Simone, Karen; Peredy, Tamas; Poli, Mark

    2014-05-01

    Paralytic shellfish poisoning (PSP), a potentially fatal foodborne illness, is often diagnosed anecdotally based on symptoms and dietary history. The neurotoxins responsible for PSP, collectively referred to as the saxitoxins or paralytic shellfish toxins (PSTs), are natural toxins, produced by certain dinoflagellates, that may accumulate in seafood, particularly filter-feeding bivalves. Illnesses are rare because of effective monitoring programs, yet occasional poisonings occur. Rarely are contaminated food and human clinical samples (e.g., urine and serum) available for testing. There are currently few methods, none of which are validated, for determining PSTs in clinical matrices. This study evaluated AOAC (Association of Analytical Communities) Official Method of Analysis (OMA) 2005.06. [AOAC Official Method 2005.06 Paralytic Shellfish Poisoning Toxins in Shellfish: Prechormatographic Oxidation and Liquid Chromatography with Fluorescence Detection. In Official Methods of Analysis of AOAC International aoac.org>], validated only for shellfish extracts, for its extension to human urine and serum samples. Initial assessment of control urine and serum matrices resulted in a sample cleanup modification when working with urine to remove hippuric acid, a natural urinary compound of environmental/dietary origin, which co-eluted with saxitoxin. Commercially available urine and serum matrices were then quantitatively spiked with PSTs that were available as certified reference materials (STX, dcSTX, B1, GTX2/3, C1/2, NEO, and GTX1/4) to assess method performance characteristics. The method was subsequently applied successfully to a PSP case study that occurred in July 2007 in Maine. Not only were PSTs identified in the patient urine and serum samples, the measured time series also led to the first report of human PST-specific urinary elimination rates. The LC-FD data generated from this case study compared remarkably well to results obtained using AOAC OMA 2011.27 [AOAC

  11. Composition and biogas yield of a novel source segregation system for pig excreta

    NARCIS (Netherlands)

    Vu, Phuong T.; Melse, Roland W.; Zeeman, Grietje; Groot Koerkamp, Peter W.G.

    2016-01-01

    The performance of a novel source segregation method for pig excreta (a V-shaped conveyor belt underneath the slatted pen floor) was compared to conventional separation methods for pig slurry (screw press, centrifugation, flocculation with/without centrifugation). For the source segregation

  12. Composition and biogas yield of a novel source segregation system for pig excreta

    NARCIS (Netherlands)

    Vu, Phuong T.; Melse, Roland W.; Zeeman, Grietje; Groot Koerkamp, Peter W.G.

    2016-01-01

    The performance of a novel source segregation method for pig excreta (a V-shaped conveyor belt underneath the slatted pen floor) was compared to conventional separation methods for pig slurry (screw press, centrifugation, flocculation with/without centrifugation). For the source segregation syste

  13. Role of excreta in predator avoidance by the Kanzawa spider mite, Tetranychus kanzawai (Acari: Tetranychidae)

    NARCIS (Netherlands)

    Oku, K.

    2008-01-01

    The Kanzawa spider mite, Tetranychus kanzawai (Acari: Tetranychidae) constructs webs over leaf surfaces and usually lives under these webs. T. kanzawai produces two types of excreta, black and yellow pellets, and uses its webs as a place for excretion. T. kanzawai also uses its webs as a refuge when

  14. Protective Effect of Distillate and Redistillate of Cow's Urine in Human Polymorphonuclear Leukocytes Challenged With Established Genotoxic Chemicals

    Institute of Scientific and Technical Information of China (English)

    K. KRISHNAMURTHI; DIPANWITA DUTTA; S. D. SIVANESAN; T. CHAKRABARTI

    2004-01-01

    From the ancient period cow's urine has been used as a medicine. In Veda, cow's urine was compared to the nectar. In Susrut, several medicinal properties of cow's urine have been mentioned and are known to cause weight loss, reversal of certain cardiac and kidney problems, indigestion, stomach ache, edema, etc. However, the literature and scripture did not mention the antigenotoxic properties of cow's urine. Methods In the present investigation, the antigenotoxic/ antioxidant properties of cow's urine distillate and redistillate were studied in vitro. The antioxidant status and volatile fatty acid levels were determined. Actinomycin-D (0.1 μmol/L) and hydrogen peroxide (150 μmol/L) were used for inducing DNA strand break with 0.1% DMSO as negative control. Dose for the antigenotoxic effect of cow's urine was chosen from the dose response study carried out earlier. Results Both actinomycin-D and H2O2 caused statistically significant DNA unwinding of 80% & 75% respectively (P<0.001) as revealed by fluorimetric analysis of DNA unwinding (FADU), and the damage could be protected with the redistilled cow's urine distillate (1, 50 & 100 μL) in simultaneous treatment with genotoxic chemicals. Conclusion The redistillate of cow's urine was found to possess total antioxidant status of around 2.6 mmol, contributed mainly by volatile fatty acids (1500 mg/L) as revealed by the GC-MS studies. These fatty acids and other antioxidants might cause the observed protective effects.

  15. The asymptomatic bacteriuria Escherichia coli strain 83972 outcompetes uropathogenic E. coli strains in human urine

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Ulett, G.C.; Schembri, M.A.

    2006-01-01

    Escherichia coli is the most common organism associated with asymptomatic bacteriuria (ABU). In contrast to uropathogenic E. coli (UPEC), which causes symptomatic urinary tract infections (UTI), very little is known about the mechanisms by which these strains colonize the human urinary tract...

  16. Flavonoids in human urine as biomarkers for intake of fruits and vegetables

    DEFF Research Database (Denmark)

    Nielsen, Salka E.; Freese, R.; Kleemola, P.

    2002-01-01

    Flavonoids are polyphenolic compounds ubiquitously found in human diets. We have studied the association between urinary excretion of flavonoids and the intake of fruits and vegetables to evaluate the usefulness of flavonoids as a biomarker for fruit and vegetable intake. Levels of 12 dietary rel...

  17. Analysis of inflammatory cytokines in human blood, breath condensate, and urine using a multiplex immunoassay platform

    Science.gov (United States)

    A change in the expression of cytokines in human biological media indicates an inflammatory response to external stressors and reflects an early step along the adverse outcome pathway (AOP) for various health endpoints. To characterize and interpret this inflammatory response, m...

  18. Determination of glucuronide conjugates of hydroxyl triphenyl phosphate (OH-TPHP) metabolites in human urine and its use as a biomarker of TPHP exposure.

    Science.gov (United States)

    Su, Guanyong; Letcher, Robert J; Yu, Hongxia; Gooden, David M; Stapleton, Heather M

    2016-04-01

    In vitro studies using avian hepatocytes or human liver microsomes suggest that hydroxylation is an important pathway in the metabolism of triphenyl phosphate (TPHP), a chemical used as a flame retardant and plasticizer. TPHP metabolism can lead to the formation of para(p)- and meta(m)-hydroxyl-(OH-)TPHP products as well as their glucuronide conjugates. To determine whether the TPHP hydroxylation and depuration pathway also occurs in vivo in humans, the present study developed a sensitive method for quantification of p- and m-OH-TPHP glucuronides in human urine samples. In n = 1 pooled urine sample and n = 12 individual urine samples collected from four human volunteers from Ottawa (ON, Canada), p- and m-OH-TPHP glucuronides were detectable in 13 and 9 of the 13 analyzed samples and at concentrations ranging from humans, and further suggests that p-OH-TPHP glucuronide can be used as a specific biomarker of TPHP exposure in humans.

  19. Urine culture - catheterized specimen

    Science.gov (United States)

    Culture - urine - catheterized specimen; Urine culture - catheterization; Catheterized urine specimen culture ... urinary tract infections may be found in the culture. This is called a contaminant. You may not ...

  20. The asymptomatic bacteriuria Escherichia coli strain 83972 outcompetes uropathogenic E. coli strains in human urine

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Ulett, G.C.; Schembri, M.A.

    2006-01-01

    Escherichia coli is the most common organism associated with asymptomatic bacteriuria (ABU). In contrast to uropathogenic E. coli (UPEC), which causes symptomatic urinary tract infections (UTI), very little is known about the mechanisms by which these strains colonize the human urinary tract....... The prototype ABU E. coli strain 83972 was originally isolated from a girl who had carried it asymptomatically for 3 years. Deliberate colonization of UTI-susceptible individuals with E. coli 83972 has been used successfully as an alternative approach for the treatment of patients who are refractory...... to conventional therapy. Colonization with strain 83972 appears to prevent infection with UPEC strains in such patients despite the fact that this strain is unable to express the primary adhesins involved in UTI, viz. P and type 1 fimbriae. Here we investigated the growth characteristics of E. coli 83972 in human...

  1. Sustitución de alimento comercial por excretas en la dieta de conejos en crecimiento

    Directory of Open Access Journals (Sweden)

    S. P. Castillo-Rodríguez

    2007-01-01

    Full Text Available La presente investigación se llevó a cabo para evaluar el efecto de la sustitución de alimento comercial por una mezcla de excretas [gallinas, cerdos y vacas] sobre el crecimiento de conejos Nueva Zelanda. Se utilizaron 30 animales con un promedio de peso de 600 + 18.2 g, y edad de 35 días, los cuales se colocaron en jaulas individuales. Los tratamientos consistieron en la sustitución parcial del alimento comercial en niveles de 0, 15, 20, 25, 30 y 35%, con una mezcla de excretas (gallinaza, cerdaza y bovinaza. La prueba tuvo una duración de 42 días, el alimento y agua de bebida fueron ofrecidos ad libitum. Todos los días se pesó el alimento ofrecido y el rechazado; el peso de los animales se registró semanalmente. Las medias generales ± desviación estándar para ganancia de peso, consumo de alimento y digestibilidad aparente fueron 866 ± 129 g, 3.02 ± 0.35 kg y 64.0 ± 0.5%, respectivamente. La ganancia de peso, consumo de alimento y digestibilidad aparente fueron afectadas por el nivel de excretas en la ración (P 0.05, pero la digestibilidad aparente fue menor en los niveles de 30 y 35% de excretas. Se puede concluir que las excretas pueden ser usadas hasta en un 25% con resultados satisfactorios y sin efectos adversos sobre la ganancia de peso y la conversión alimenticia.

  2. Comparison study of the sensitivities of some indices of DDT exposure in human blood and urine

    Energy Technology Data Exchange (ETDEWEB)

    Nhachi, C.F.B.; Loewenson, R. (Univ. of Zimbabwe (South Africa))

    1989-10-01

    Although exposure to DDT (2,2-bis(p-chlorophenyl1)1,1,1,-trichloroethane) is not normally associated with fatality or chronic adverse effects to human life, it is a known hazard to the ecosystem. Blood levels of DDT and some of its derivatives have been used to assess extent of exposure or the body load of DDT in humans. In experimental studies, ingestion of DDT has been associated with reduced liver stores of vitamin A, and increased serum levels of vitamin A. The same study also revealed a significant correlation of vitamin A and DDE serum levels. Generally an increase in excreted 17-B-hydroxycortisone has been associated with DDT exposure. Increased excretion of 6-B-hydroxycortisol has been noted in workers who were involved in the formulation of DDT. The objective of this study was to compare the sensitivities of some indices of DDT exposure in humans. The indices which were compared are serum vitamin A and DDE levels and urinary 17-B-hydroxycortisol.

  3. Capillary electrophoresis-time of flight-mass spectrometry using noncovalently bilayer-coated capillaries for the analysis of amino acids in human urine.

    Science.gov (United States)

    Ramautar, Rawi; Mayboroda, Oleg A; Derks, Rico J E; van Nieuwkoop, Cees; van Dissel, Jaap T; Somsen, Govert W; Deelder, André M; de Jong, Gerhardus J

    2008-06-01

    A capillary electrophoresis-time of flight-mass spectrometry (CE-TOF-MS) method for the analysis of amino acids in human urine was developed. Capillaries noncovalently coated with a bilayer of Polybrene (PB) and poly(vinyl sulfonate) (PVS) provided a considerable EOF at low pH, thus facilitating the fast separation of amino acids using a BGE of 1 M formic acid (pH 1.8). The PB-PVS coating proved to be very consistent yielding stable CE-MS patterns of amino acids in urine with favorable migration time repeatability (RSDs capillary preconcentration step based on pH-mediated stacking allowing 100-nL sample injection (i.e. ca. 4% of capillary volume). As a result, LODs for amino acids were down to 20 nM while achieving satisfactory separation efficiencies. Preliminary validation of the method with urine samples showed good linear responses for the amino acids (R(2) >0.99), and RSDs for peak areas were <10%. Special attention was paid to the influence of matrix effects on the quantification of amino acids. The magnitude of ion suppression by the matrix was similar for different urine samples. The CE-TOF-MS method was used for the analysis of urine samples of patients with urinary tract infection (UTI). Concentrations of a subset of amino acids were determined and compared with concentrations in urine of healthy controls. Furthermore, partial least squares-discriminant analysis (PLS-DA) of the CE-TOF-MS dataset in the 50-450 m/z region showed a distinctive grouping of the UTI samples and the control samples. Examination of score and loadings plot revealed a number of compounds, including phenylalanine, to be responsible for grouping of the samples. Thus, the CE-TOF-MS method shows good potential for the screening of body fluids based on the analysis of endogenous low-molecular weight metabolites such as amino acids and related compounds.

  4. Electroanalytical Determination of Gemifloxacin Mesylate in Bulk, Tablets and Human Urine Using Gold Nanoparticles Modified Carbon Paste Electrode

    Directory of Open Access Journals (Sweden)

    Ali Attia

    2014-12-01

    Full Text Available A simple, precise, inexpensive and sensitive voltammetric method has been developed for the determination of gemifloxacin mesylate (GEM in the presence of tween 80 in the bulk, farmaceutical dosage forms and human urine at gold nanoparticles modified carbon paste electrode (GNCPE. The electrochemical behavior of GEM has been investigated by using cyclic voltammetry (CV and differential pulse voltammetry (DPV techniques. The electrochemical oxidation of GEM was an irreversible process which exhibited adsorption-diffusion controlled process behavior in Britton-Robinson (BR buffer over the entire pH range of values from 2 to 9. The adsorptive stripping response was evaluated as a function of some variables such as pH, type of surfactant, scan rate and accumulation time. The anodic peak current varied linearly over the range from 8.0 × 10-7 to 2.8 × 10-5 M. The limits of detection and quantification were 7.32 × 10-8 M and 2.44 × 10-7 M, respectively. The relative standard deviations and the percentage recoveries were found in the following ranges: 0.58-1.35% and 99.37-101.76%, respectively.

  5. A rapid and sensitive resonance Rayleigh scattering spectra method for the determination of quinolones in human urine and pharmaceutical preparation.

    Science.gov (United States)

    Qiao, Man; Wang, Yaqiong; Liu, Shaopu; Liu, Zhongfang; Yang, Jidong; Zhu, Jinghui; Hu, Xiaoli

    2015-03-01

    A new method based on resonance Rayleigh scattering (RRS) was proposed for the determination of quinolones (QNS) at the nanogram level. In pH 3.3-4.4 Britton-Robinson buffer medium, quinolones such as ciprofloxacin, pipemidic acid (PIP), lomefloxacin (LOM), norfloxacin (NOR) and sarafloxacin (SAR) were protonated and reacted with methyl orange (MO) to form an ion-pair complex, which then further formed a six-membered ring chelate with Pd(II). As a result, new RRS spectra appeared and the RRS intensities were enhanced greatly. RRS spectral characteristics of the MO-QNS-Pd(II) systems, the optimum conditions for the reaction, and the influencing factors were investigated. Under optimum conditions, the scattering intensity (∆I) increments were directly proportional to the concentration of QNS with in certain ranges. The method had high sensitivity, and the detection limits (3σ) ranged from 6.8 to 12.6 ng/mL. The proposed method had been successfully applied for the determination of QNS in pharmaceutical formulations and human urine samples. In addition, the mechanism of the reaction system was discussed based on IR, absorption and fluorescence spectral studies. The reasons for the enhancement of scattering spectra were discussed in terms of fluorescence-scattering resonance energy transfer, hydrophobicity and molecular size.

  6. Ionophore-Based Potentiometric Sensors for the Flow-Injection Determination of Promethazine Hydrochloride in Pharmaceutical Formulations and Human Urine

    Directory of Open Access Journals (Sweden)

    Suad Mustafa Al-Araji

    2011-01-01

    Full Text Available Plasticised poly(vinyl chloride-based membranes containing the ionophores (α-, β- and γ-cyclodextrins (CD, dibenzo-18-crown-6 (DB18C6 and dibenzo-30-crown-10 (DB30C10 were evaluated for their potentiometric response towards promethazine (PM in a flow injection analysis (FIA set-up. Good responses were obtained when β- and γ-CDs, and DB30C10 were used. The performance characteristics were further improved when tetrakis(4-chlorophenyl borate (KTPB was added to the membrane. The sensor based on β-CD, bis(2-ethylhexyl adipate (BEHA and KTPB exhibited the best performance among the eighteen sensor compositions that were tested. The response was linear from 1 x 10−5 to 1 x 10−2 M, slope was 61.3 mV decade−1, the pH independent region ranged from 4.5 to 7.0, a limit of detection of 5.3 x 10−6 M was possible and a lifetime of more than a month was observed when used in the FIA system. Other plasticisers such as dioctyl phenylphosphonate and tributyl phosphate do not show significant improvements in the quality of the sensors. The promising sensors were further tested for the effects of foreign ions (Li+, Na+, K+, Mg2+, Ca2+, Co2+, Cu2+, Cr3+, Fe3+, glucose, fructose. FIA conditions (e.g., effects of flow rate, injection volume, pH of the carrier stream were also studied when the best sensor was used (based on β-CD. The sensor was applied to the determination of PM in four pharmaceutical preparations and human urine that were spiked with different levels of PM. Good agreement between the sensor and the manufacturer’s claimed values (for pharmaceutical preparations was obtained, while mean recoveries of 98.6% were obtained for spiked urine samples. The molecular recognition features of the sensors as revealed by molecular modelling were rationalised by the nature of the interactions and complexation energies between the host and guest molecules.

  7. Detection and genotyping of human papillomavirus in urine samples from unvaccinated male and female adolescents in Italy.

    Science.gov (United States)

    Bianchi, Silvia; Frati, Elena Rosanna; Panatto, Donatella; Martinelli, Marianna; Amicizia, Daniela; Zotti, Carla Maria; Martinese, Morena; Bonanni, Paolo; Boccalini, Sara; Coppola, Rosa Cristina; Masia, Giuseppina; Meloni, Angelo; Castiglia, Paolo; Piana, Andrea; Gasparini, Roberto; Tanzi, Elisabetta

    2013-01-01

    The introduction of vaccination against Human Papillomavirus (HPV) in adolescent girls in 2006 has focused virological surveillance on this age group. As few studies have evaluated HPV infections in young populations, further data are needed in order to improve and extend prophylactic policy and to monitor epidemiological changes. The present study aimed at evaluating overall and type-specific HPV prevalence in both female and male adolescents in Italy. HPV DNA detection and genotyping was performed on urine samples collected from 870 unvaccinated adolescents (369 females, 501 males, 11-18 years of age) in five cities in Italy. Following DNA extraction by means of a commercial kit (NucliSENS(®)-miniMAG(®), bioMérieux), the L1 gene fragment was PCR amplified and genotyped by restriction fragment length polymorphism analysis. HPV DNA was detected in 1.5% of all samples, and in 3% and 0.4% of samples from females and males, respectively. In approximately 70% of HPV DNA positive adolescents, the infection was due to a single genotype, with 88.9% of genotypes belonging to the HR-clade. The only two HPV-positive boys (14 and 18 years old) had HPV-70 genotype. Only one of the 11 HPV-infected girls was in the 11-14 age-group. HPV prevalence was 4.2% in girls aged 15-18 years and 60% of infections were due to vaccine types HPV-16 or HPV-6/-11. This is one of the few studies, the first conducted in Italy, on HPV infection in adolescents. Urine testing is the easier way of detecting HPV infection in younger populations. Our data revealed a very low HPV prevalence, and no infections were observed in the 12-year-old vaccine target population. The majority of infections were seen in females aged 15-18 years. Overall, more than 50% and 30% of the potentially persistent HPV infections detected in this group could have been prevented by the quadrivalent and the bivalent vaccines, respectively.

  8. Detection and genotyping of human papillomavirus in urine samples from unvaccinated male and female adolescents in Italy.

    Directory of Open Access Journals (Sweden)

    Silvia Bianchi

    Full Text Available The introduction of vaccination against Human Papillomavirus (HPV in adolescent girls in 2006 has focused virological surveillance on this age group. As few studies have evaluated HPV infections in young populations, further data are needed in order to improve and extend prophylactic policy and to monitor epidemiological changes. The present study aimed at evaluating overall and type-specific HPV prevalence in both female and male adolescents in Italy. HPV DNA detection and genotyping was performed on urine samples collected from 870 unvaccinated adolescents (369 females, 501 males, 11-18 years of age in five cities in Italy. Following DNA extraction by means of a commercial kit (NucliSENS(®-miniMAG(®, bioMérieux, the L1 gene fragment was PCR amplified and genotyped by restriction fragment length polymorphism analysis. HPV DNA was detected in 1.5% of all samples, and in 3% and 0.4% of samples from females and males, respectively. In approximately 70% of HPV DNA positive adolescents, the infection was due to a single genotype, with 88.9% of genotypes belonging to the HR-clade. The only two HPV-positive boys (14 and 18 years old had HPV-70 genotype. Only one of the 11 HPV-infected girls was in the 11-14 age-group. HPV prevalence was 4.2% in girls aged 15-18 years and 60% of infections were due to vaccine types HPV-16 or HPV-6/-11. This is one of the few studies, the first conducted in Italy, on HPV infection in adolescents. Urine testing is the easier way of detecting HPV infection in younger populations. Our data revealed a very low HPV prevalence, and no infections were observed in the 12-year-old vaccine target population. The majority of infections were seen in females aged 15-18 years. Overall, more than 50% and 30% of the potentially persistent HPV infections detected in this group could have been prevented by the quadrivalent and the bivalent vaccines, respectively.

  9. Cellular characterization of human dermal fibroblasts, focus on mitochondria and maple syrup urine disease

    DEFF Research Database (Denmark)

    Fernandez-Guerra, Paula

    and functions are expressed in HDFs’ culture environment. Studies of molecular disease mechanisms often point to the involvement of mitochondria. Mitochondria are involved in the regulation of cell cycle and programmed cell death as well as cellular stress responses because they are the main producers......Cell phenotyping of human dermal fibroblasts (HDFs) from patients with inherited metabolic diseases (IMDs) provide invaluable information for diagnosis, disease aetiology, predicting prognosis, and monitoring of treatments. HDFs possess the genetic composition of patients and many pathways...... of reactive oxygen species (ROS). Advances in technology help the study of complex situations with large amount of data, like cellular phenotyping in cell culture. Image cytometry is an emerging technique that combines morphological information and fluorescent intensity data from single cells. We defined...

  10. Urine Tests (For Parents)

    Science.gov (United States)

    ... TOPIC Vesicoureteral Reflux (VUR) Blood in the Urine (Hematuria) Urine Test: Creatinine Urine Test: Microalbumin-to-Creatinine ... Video) Urinary Tract Infections Blood in the Urine (Hematuria) Kidneys and Urinary Tract Contact Us Print Resources ...

  11. Clean catch urine sample

    Science.gov (United States)

    ... opening of the vagina. To collect the urine sample: Keeping your labia spread open, urinate a small amount into the toilet bowl, then stop the flow of urine. Hold the urine cup a few inches (or a ...

  12. Urination - difficulty with flow

    Science.gov (United States)

    ... common cause is infection of the prostate or urinary tract. Symptoms of a possible infection include: Burning or pain with urination Frequent urination Cloudy urine Sense of urgency (strong, sudden urge to urinate) The problem can ...

  13. Angiotensin converting enzymes from human urine of mild hypertensive untreated patients resemble the N-terminal fragment of human angiotensin I-converting enzyme.

    Science.gov (United States)

    Casarini, D E; Plavinik, F L; Zanella, M T; Marson, O; Krieger, J E; Hirata, I Y; Stella, R C

    2001-01-01

    Angiotensin I-converting enzyme (ACE) activity was analyzed in human urine collected from mild hypertensive untreated patients. DEAE-cellulose chromatography using linear gradient elution revealed two forms of angiotensin I-converting enzyme, eluted in the conductivity of 0.75 and 1.25 mS. The fractions of each conductivity were pooled and submitted to direct gel filtration in an AcA-34 column, and the apparent molecular weights of urinary ACEs were estimated as 90 kDa (for ACE eluted in 0.75 mS) and 65 kDa (for ACE eluted in 1.25 mS). Both enzymes have a K(i) of the order of 10(-7) M for the specific inhibitors studied, and are able to hydrolyze luteinizing hormone-releasing hormone and N-acetyl-Ser-Asp-Lys-Pro as described for N-domain ACE. By Western blot analysis, both peaks were recognized by ACE-specific antibody Y4, confirming the molecular weight already described. A plate precipitation assay using monoclonal antibodies to the N-domain of ACE showed that both forms of ACE binds with all monoclonal antibodies to the active N-domain ACE, suggesting that these forms of human urine ACEs resemble the N-fragment of ACE. The HP2 ACE (65 kDa) is similar to low molecular weight (LMW) ACE from normal subjects, and the HP2 ACE (90 kDa) is different from high molecular weight (190 kDa) and LMW (65 kDa) normal ACEs. The 90 kDa ACE could have an important role in development of hypertension. It will be fundamental to elucidate the molecular mechanism responsible for the genesis of this isoform.

  14. Simultaneous determination of neonicotinoid insecticides in human serum and urine using diatomaceous earth-assisted extraction and liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Yamamuro, Tadashi; Ohta, Hikoto; Aoyama, Mika; Watanabe, Daisuke

    2014-10-15

    A rapid and sensitive analytical method was developed for simultaneous determination of eight neonicotinoid insecticides (acetamiprid, clothianidin, dinotefuran, flonicamid, imidacloprid, nitenpyram, thiacloprid and thiamethoxam) and three specific metabolites of acetamiprid (N-desmethylacetamiprid, 5-(N-acetyl-N-methylaminomethyl)-2-chloropyridine and 5-(N-acetylaminomethyl)-2-chloropyridine) in human serum and urine. A diatomaceous earth-assisted extraction using Extrelut NT3 column with chloroform/2-propanol (3:1, v/v) as eluent was selected for the single step cleanup procedure for all the target compounds. Qualitative and quantitative analyses were conducted by liquid chromatography-tandem mass spectrometry with multiple reaction monitoring mode. The limits of detection and the limits of quantification of eleven compounds were in the ranges of 0.1-0.2ng/mL and 0.5-10ng/mL for serum, 0.1-1ng/mL and 1-10ng/mL for urine, respectively. The extraction recoveries were between 80.9% and 101.8% for serum samples, 91.9% and 106% for urine samples. The intra-day RSDs and the inter-day RSDs were less than 11.5% and 13.2% for serum, less than 8.3% and 8.8% for urine. The proposed procedure will be suitable for forensic investigations of human poisoning cases with neonicotinoid insecticides. This is the first report of simultaneous determination of eight neonicotinoids in serum and urine samples. Copyright © 2014. Published by Elsevier B.V.

  15. Determination of parabens in human urine by optimal ultrasound-assisted emulsification microextraction and on-line acetylation gas chromatography-mass spectrometry.

    Science.gov (United States)

    Hui-Ting, Zhou; Ding, Erica M C; Ding, Wang-Hsien

    2017-07-15

    An effective and solvent-less method for the rapid determination of four commonly detected parabens (methyl-, ethyl-, propyl- and butyl-) in human urine samples is described. This method employed ultrasound-assisted emulsification microextraction (USAEME) before identification and quantitation of the parabens via on-line acetylation gas chromatography-mass spectrometry (GC-MS). Urine samples were enzymatically de-conjugated with β-glucuronidase and then extracted by an optimal USAEME procedure for the measurement of total concentrations of target analytes. The optimal USAEME parameters for one mL of urine sample (containing 0.1-g of sodium chloride), according to the Box-Behnken design method, are thus described: extractant of 200-μL of ethyl acetate, and ultrasonication for 1.0min and centrifugation at 7000rpm (3min). The supernatant was collected and evaporated until dry. Then the residue was re-dissolved in methanol (100-μL), and the extract was subjected to on-line acetylation GC-MS analysis. The limits of quantitation (LOQs) were less than 0.06ng/mL. Precisions for both intra- and inter-day analysis were calculated, and were less than 8%. Mean extraction recovery (known as trueness) was between 83 and 101% on three concentration levels. In human urine, the total concentrations of the four selected parabens, according to preliminary results, range from 0.3 to 124.5ng/mL for male, and from 27.2 to 246.3ng/mL for female. Female urine samples showed higher concentrations for the target parabens, which may indicate higher exposure due to lifestyle. This method permits accurate and high-throughput analysis of parabens for epidemiological studies. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Liquid chromatography-tandem mass spectrometry (LC/APCI-MS/MS) methods for the quantification of captan and folpet phthalimide metabolites in human plasma and urine.

    Science.gov (United States)

    Berthet, Aurélie; Bouchard, Michèle; Schüpfer, Patrick; Vernez, David; Danuser, Brigitta; Huynh, Cong Khanh

    2011-02-01

    Captan and folpet are fungicides largely used in agriculture. They have similar chemical structures, except that folpet has an aromatic ring unlike captan. Their half-lives in blood are very short, given that they are readily broken down to tetrahydrophthalimide (THPI) and phthalimide (PI), respectively. Few authors measured these biomarkers in plasma or urine, and analysis was conducted either by gas chromatography coupled to mass spectrometry or liquid chromatography with UV detection. The objective of this study was thus to develop simple, sensitive and specific liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry (LC/APCI-MS/MS) methods to quantify both THPI and PI in human plasma and urine. Briefly, deuterated THPI was added as an internal standard and purification was performed by solid-phase extraction followed by LC/APCI-MS/MS analysis in negative ion mode for both compounds. Validation of the methods was conducted using spiked blank plasma and urine samples at concentrations ranging from 1 to 250 μg/L and 1 to 50 μg/L, respectively, along with samples of volunteers and workers exposed to captan or folpet. The methods showed a good linearity (R (2) > 0.99), recovery (on average 90% for THPI and 75% for PI), intra- and inter-day precision (RSD, <15%) and accuracy (<20%), and stability. The limit of detection was 0.58 μg/L in urine and 1.47 μg/L in plasma for THPI and 1.14 and 2.17 μg/L, respectively, for PI. The described methods proved to be accurate and suitable to determine the toxicokinetics of both metabolites in human plasma and urine.

  17. Treating urine by Spirulina platensis

    Science.gov (United States)

    Yang, Chenliang; Liu, Hong; Li, Ming; Yu, Chengying; Yu, Gurevich

    In this paper Spirulina platensis with relatively high nutrition was cultivated to treat human urine. Batch culture showed that the consumption of N in human urine could reach to 99%, and the consumption of P was more than 99.9%, and 1.05 g biomass was obtained by treating 12.5 ml synthetic human urine; continuous culture showed that S. platensis could consume N, Cl, K and S in human urine effectively, and the consumption could reach to 99.9%, 75.0%, 83.7% and 96.0%, respectively, and the consumption of P was over 99.9%, which is very important to increase the closure and safety of the bioregenerative life support system (BLSS).

  18. Separation and identification of Se-methylselenogalactosamine - a new metabolite in basal human urine - by HPLC-ICP-MS and CE-nano-ESI-(MS)(2)

    DEFF Research Database (Denmark)

    Bendahl, L.; Gammelgaard, Bente

    2004-01-01

    Three minor metabolites were isolated from human urine. Two of these were identified by nano electrospray ionisation mass spectrometry (nESI-MS) as Se-methylseleno-N-acetylglucosamine and Se-methylselenogalactosamine, respectively. A human urine pool was lyophilised and reconstituted in methanol....... It was not possible to identify this selenium compound on basis of the available data. The selenium compound in the second fraction showed co-elution with a Se-methylseleno-N-acetylglucosamine standard. The identity of this compound was verified by nESI-MS after further purification by size exclusion chromatography....... The third fraction was further purified by ion-pair and anion exchange chromatography, reconstituted and subjected to CE-nESI-MS. The m/z of the compound was 258 and ( MS) 2 resulted in a fragment at m/z 162, corresponding to loss of methylselenium. This indicated that the structure of the compound was Se...

  19. Accurate quantification of the mercapturic acids of acrylonitrile and its genotoxic metabolite cyanoethylene-epoxide in human urine by isotope-dilution LC-ESI/MS/MS.

    Science.gov (United States)

    Schettgen, T; Bertram, J; Kraus, T

    2012-08-30

    Acrylonitrile is a highly important industrial chemical with a high production volume worldwide, especially in the plastics industry. It is classified as a possible human carcinogen by the International Agency for Research on Cancer (IARC group 2B). During metabolism of acrylonitrile, the genotoxic metabolite cyanoethylene-epoxide is formed. The urinary mercapturic acids of acrylonitrile, namely N-acetyl-S-(2-cyanoethyl)-L-cysteine (CEMA) and cyanoethylene-epoxide, namely N-acetyl-S-(1-cyano-2-hydroxyethyl)-L-cysteine (CHEMA) are specific biomarkers for the determination of individual internal exposure to acrylonitrile and its highly reactive metabolite. We have developed and validated a sensitive method for the accurate determination of CEMA and CHEMA in human urine with a multidimensional LC/MS/MS-method using deuterium-labelled analogues for both analytes as internal standards. Analytes were stripped from urinary matrix by online extraction on a restricted access material, transferred to the analytical column and determined by tandem mass spectrometry. The limit of quantification (LOQ) for CEMA and CHEMA was 1 μg/L urine and allowed to quantify the background exposure of the (smoking) general population. Precision within and between series for CHEMA ranged from 2.6 to 8.0% at four concentrations ranging from 8.3 to 86 μg/L urine, mean accuracy was between 94 and 100%. For CEMA, precision within and between series ranged from 2.4 to 14.5% at four concentrations ranging from 15.1 to 196 μg/L urine, mean accuracy was between 91 and 104%. We applied the method to spot urine samples of 83 subjects of the general population with no known occupational exposure to acrylonitrile. Median levels (range) for CEMA and CHEMA in urine samples of non-smokers (n=47) were 1.9 μg/L (<1-16.4 μg/L) and<1 μg/L (<1-3 μg/L), while in urine samples of smokers (n=36), median levels were 184 μg/L (2-907 μg/L) and 29.3 μg/L (<1-147 μg/L), respectively. Smokers showed a

  20. Validation of a method to quantify chromium, cadmium, manganese, nickel and lead in human whole blood, urine, saliva and hair samples by electrothermal atomic absorption spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Olmedo, P.; Pla, A.; Hernandez, A.F.; Lopez-Guarnido, O.; Rodrigo, L. [Department of Legal Medicine and Toxicology, University of Granada, School of Medicine (Spain); Gil, F., E-mail: fgil@ugr.es [Department of Legal Medicine and Toxicology, University of Granada, School of Medicine (Spain)

    2010-02-05

    For biological monitoring of heavy metal exposure in occupational toxicology, usually whole blood and urine samples are the most widely used and accepted matrix to assess internal xenobiotic exposure. Hair samples and saliva are also of interest in occupational and environmental health surveys but procedures for the determination of metals in saliva and hair are very scarce and to our knowledge there is no validation of a method to quantify Cr, Cd, Mn, Ni and Pb in four different human biological materials (whole blood, urine, saliva and axilary hair) by electrothermal atomization atomic absorption spectrometry (ETAAS). In the present study, quantification methods for the determination of Cr, Cd, Mn, Ni and Pb in whole blood, urine, saliva and axilary hair were validated according to the EU common standards. Pyrolisis and atomization temperatures have been determined. The main parameters evaluated were: detection and quantification limits, linearity range, repeatability, reproducibility, recovery and uncertainty. Accuracy of the methods was tested with the whole blood, urine and hair certified reference materials and recoveries of the spiked samples were acceptable ranged from 96.3 to 107.8%.

  1. Validation of a method to quantify chromium, cadmium, manganese, nickel and lead in human whole blood, urine, saliva and hair samples by electrothermal atomic absorption spectrometry.

    Science.gov (United States)

    Olmedo, P; Pla, A; Hernández, A F; López-Guarnido, O; Rodrigo, L; Gil, F

    2010-02-05

    For biological monitoring of heavy metal exposure in occupational toxicology, usually whole blood and urine samples are the most widely used and accepted matrix to assess internal xenobiotic exposure. Hair samples and saliva are also of interest in occupational and environmental health surveys but procedures for the determination of metals in saliva and hair are very scarce and to our knowledge there is no validation of a method to quantify Cr, Cd, Mn, Ni and Pb in four different human biological materials (whole blood, urine, saliva and axilary hair) by electrothermal atomization atomic absorption spectrometry (ETAAS). In the present study, quantification methods for the determination of Cr, Cd, Mn, Ni and Pb in whole blood, urine, saliva and axilary hair were validated according to the EU common standards. Pyrolisis and atomization temperatures have been determined. The main parameters evaluated were: detection and quantification limits, linearity range, repeatability, reproducibility, recovery and uncertainty. Accuracy of the methods was tested with the whole blood, urine and hair certified reference materials and recoveries of the spiked samples were acceptable ranged from 96.3 to 107.8%.

  2. Monoclonal antibody-based dipstick assay: a reliable field applicable technique for diagnosis of Schistosoma mansoni infection using human serum and urine samples.

    Science.gov (United States)

    Demerdash, Zeinab; Mohamed, Salwa; Hendawy, Mohamed; Rabia, Ibrahim; Attia, Mohy; Shaker, Zeinab; Diab, Tarek M

    2013-02-01

    A field applicable diagnostic technique, the dipstick assay, was evaluated for its sensitivity and specificity in diagnosing human Schistosoma mansoni infection. A monoclonal antibody (mAb) against S. mansoni adult worm tegumental antigen (AWTA) was employed in dipstick and sandwich ELISA for detection of circulating schistosome antigen (CSA) in both serum and urine samples. Based on clinical and parasitological examinations, 60 S. mansoni-infected patients, 30 patients infected with parasites other than schistosomiasis, and 30 uninfected healthy individuals were selected. The sensitivity and specificity of dipstick assay in urine samples were 86.7% and 90.0%, respectively, compared to 90.0% sensitivity and 91.7% specificity of sandwich ELISA. In serum samples, the sensitivity and specificity were 88.3% and 91.7% for dipstick assay vs. 91.7% and 95.0% for sandwich ELISA, respectively. The diagnostic efficacy of dipstick assay in urine and serum samples was 88.3% and 90.0%, while it was 90.8% and 93.3% for sandwich ELISA, respectively. The diagnostic indices of dipstick assay and ELISA either in serum or in urine were statistically comparable (P>0.05). In conclusion, the dipstick assay offers an alternative simple, rapid, non-invasive technique in detecting CSA or complement to stool examinations especially in field studies.

  3. Tandem focused ultrasound (TFU) combined with fast furnace analysis as an improved methodology for total mercury determination in human urine by electrothermal-atomic absorption spectrometry.

    Science.gov (United States)

    Capelo, J L; Dos Reis, C D; Maduro, C; Mota, A

    2004-09-01

    A new sample preparation procedure based on tandem (that is, different diameter probe sonicators used in the same sample treatment) focused ultrasound (TFU) for mercury separation, preconcentration and back-extraction in aqueous solution from human urine has been developed. The urine is first oxidized with KMnO(4)/HCl/focused ultrasound (6mm probe). Secondly, the mercury is extracted and preconcentrated with dithizone and cyclohexane. Finally, the mercury is back-extracted and preconcentrated again with the aid of focused ultrasound (3mm probe). The procedure allows determining mercury by electrothermal atomic absorption spectrometry with fast furnace analysis and calibration against aqueous standards. Matrix modification is provided by the chemicals used in the sample treatment. The procedure is accomplished with low sample volume (8.5ml). Low volume and low concentration reagents are used. The sample treatment is rapid (less than 3min per sample) and avoids the use of organic phase in the graphite furnace. The preconcentration factor used in this work was 14. The limit of detection and the limit of quantification in urine were, respectively, 0.27 and 0.9mugl(-1). The relative standard deviation of aqueous standards (n=10) was 4% for a concentration of 100mugl(-1) and 5% for a concentration of 400mugl(-1). Recoveries from spiked urine with inorganic mercury, methyl-mercury, phenyl-mercury and diphenyl-mercury ranged from 86 to 98%.

  4. Sequential extraction of inorganic arsenic compounds and methyl arsenate in human urine using mixed-mode monolithic silica spin column coupled with gas chromatography-mass spectrometry.

    Science.gov (United States)

    Namera, Akira; Takeuchi, Akito; Saito, Takeshi; Miyazaki, Shota; Oikawa, Hiroshi; Saruwatari, Tatsuro; Nagao, Masataka

    2012-09-01

    A sequential analytical method was developed for the detection of arsenite, arsenate, and methylarsenate in human urine by gas chromatography-mass spectrometry (GC-MS). The combination of a derivatization of trivalent arsenic compounds by 2,3-dithio-1-propanol (British antilewisite; BAL) and a reduction of pentavalent arsenic compounds (arsenate and methylarsenate) were accomplished to carry out the analysis of arsenic compounds in urine. The arsenic derivatives obtained using BAL were extracted in a stepwise manner using a monolithic spin column and analyzed by GC-MS. A linear curve was observed for concentrations of arsenic compounds of 2.0 to 200 ng/mL, where the correlation coefficients of calibration curves were greater than 0.996 (for a signal-to-noise (S/N) ratio >10). The detection limits were 1 ng/mL (S/N > 3). Recoveries of the targets in urine were in the range 91.9-106.5%, and RSDs of the intra- and interday assay for urine samples containing 5, 50, and 150 ng/mL of arsenic compounds varied between 2.95 and 13.4%. The results from real samples obtained from a patient suspected of having ingested As containing medications using this proposed method were in good agreement with those obtained using high-performance liquid chromatography with inductively coupled plasma mass spectrometry.

  5. [An LC-MS/MS method for the simultaneous determination of amygdalin and paeoniflorin in human urine and application to urinary excretion study].

    Science.gov (United States)

    Li, Xiao-bing; Shi, Fu-guo; Jian, Ling-yan; Ding, Li

    2015-10-01

    The study aims to develop an LC-MS/MS method for the simultaneous determination of amygdalin and paeoniflorin in urine samples, and to investigate their urinary excretion characteristics in healthy volunteers after intravenous infusion administration of Huoxue-Tongluo lyophilized powder for injection (HTLPI). The urine samples were extracted by methanol, and then separated on a Hedera ODS-2 column with a mobile phase of acetonitrile and 5 mmol · L(-1) ammonium acetate buffer solution containing 0.05% formic acid (20:80). Electrospray ionization source was applied and operated in the positive ion mode using MRM. The method exhibited good linearity over the concentration range of 0.03 -40 µg · mL(-1). The values on both the occasions (intra- and inter-day) were all within 15% at three concentration levels. No matrix effect and carry-over effect were observed. Amygdalin and paeoniflorin were stable in human urine under different storage conditions. Approximately 79.6% of the administered amount of amygdalin was excreted unchanged in urine within 24 h and which was 48.4% for paeoniflorin. The developed LC-MS/MS method can be applied to evaluate the urinary excretion of amygdalin and paeoniflorin.

  6. Three-dimensional surface-enhanced Raman scattering hotspots in spherical colloidal superstructure for identification and detection of drugs in human urine.

    Science.gov (United States)

    Han, Zhenzhen; Liu, Honglin; Wang, Bin; Weng, Shizhuang; Yang, Liangbao; Liu, Jinhuai

    2015-01-01

    Rapid component separation and robust surface-enhanced Raman scattering (SERS) identification of drugs in real human urine remain an attractive challenge because of the sample complexity, low molecular affinity for metal surface, and inefficient use of hotspots in one- or two-dimensional (2D) geometries. Here, we developed a 5 min strategy of cyclohexane (CYH) extraction for separating amphetamines from human urine. Simultaneously, an oil-in-water emulsion method is used to assemble monodisperse Ag nanoparticles in the CYH phase into spherical colloidal superstructures in the aqueous phase. These superstructures create three-dimensional (3D) SERS hotspots which exist between every two adjacent particles in 3D space, break the traditional 2D limitation, and extend the hotspots into the third dimension along the z-axis. In this platform, a conservative estimate of Raman enhancement factor is larger than 10(7), and the same CYH extraction processing results in a high acceptability and enrichment of drug molecules in 3D hotspots which demonstrates excellent stability and reproducibility and is suitable for the quantitative examination of amphetamines in both aqueous and organic phases. Parallel ultraperformance liquid chromatography (UPLC) examinations corroborate an excellent performance of our SERS platform for the quantitative analysis of methamphetamine (MA) in both aqueous solution and real human urine, of which the detection limits reach 1 and 10 ppb, respectively, with tolerable signal-to-noise ratios. Moreover, SERS examinations on different proportions of MA and 3,4-methylenedioxymethamphetamine (MDMA) in human urine demonstrate an excellent capability of multiplex quantification of ultratrace analytes. By virtue of a spectral classification algorithm, we realize the rapid and accurate recognition of weak Raman signals of amphetamines at trace levels and also clearly distinguish various proportions of multiplex components. Our platform for detecting drugs

  7. The simultaneous identification of metoprolol and its major acidic and basic metabolites in human urine by gas chromatography-mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Li, Feng; Cooper, S.F. [Universite du Quebec, Pointe-Claire (Canada)

    1996-12-31

    A novel gas chromatography-mass spectrometric (GC-MS) method was developed to confirm and identify metoprolol and its metabolites by double derivatization with S-(-)menthyl chloroformate [(-)-MCF] and N-methyl(trimethylsilyl-trifluoroacetamide) (MSTFA). This is the first report, which describes the simultaneous identification of metoprolol, its one major acidc and other basic metabolites in human urine based on solid-phase extraction with C{sub 18} reversed-phase cartridges. 12 refs., 4 figs.

  8. A Simple and Sensitive LC-MS/MS Method for the Determination of Free 8-Hydroxy-2'-Deoxyguanosine in Human Urine

    Science.gov (United States)

    Wang, Zuwei; Smith, Scott M.

    2016-01-01

    Urinary free 8-hydroxy-2'-deoxyguanosine (8OHdG), an oxidized product of DNA, and is frequently chosen as a biomarker of oxidative stress in humans, including studies of oxidative DNA damage during space flight. It is challenging to accurately and efficiently quantify urinary free 8OHdG in large scale human studies. LC-MS/MS is emerging as a preferable analytical technique owing its high sensitivity, selectivity and efficiency, compared to some traditional methods such as ELISA and HPLC. A simple and sensitive LC-MS/MS method has been developed for the determination of free 8OHdG in human urine. Sample preparation was done by solid phase extraction with a Waters Oasis HLB 96 well plate. A Waters Alliance 2795 HT Separation Module combined with a Quattro Micro tandem mass spectrometer was used as the LC-MS/MS system. The runtime of one injection can be less than 5 minutes using a reversed phase C18 column and an isocratic flow of methanol/water. ESI positive ions were quantified in the multiple reaction modes (MRM) using m/z 284 yields 168 for 8OHdG and m/z 289 yields173 for stable isotope labeled internal standard [(15)N5] 8OHdG. With this method for 8OHdG, a lower limit of quantitation of 1.0 nM (0.28 ng/mL) has been achieved using 100 microliter urine sample. The analytical range is between 1.0 and 100 nM with a correlation coefficient greater than or equal to 0.99. Good reproducibility can be obtained with intra-assay and inter-assay CVs less than or equal to 10% for 8OHdG spiked urine QC samples. This method can be used in high-throughput routine analysis of free 8OHdG in human urine.

  9. Sensitive determination of parabens in human urine and serum using methacrylate monoliths and reversed-phase capillary liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Carrasco-Correa, Enrique Javier; Vela-Soria, Fernando; Ballesteros, Oscar; Ramis-Ramos, Guillermo; Herrero-Martínez, José Manuel

    2015-01-30

    A method for the determination of parabens in human urine and serum by capillary liquid chromatography (cLC) with UV-Vis and mass spectrometry (MS) detection using methacrylate ester-based monolithic columns has been developed. The influence of composition of polymerization mixture was studied. The optimum monolith was obtained with butyl methacrylate monomer at 60/40% (wt/wt) butyl methacrylate/ethylene dimethacrylate ratio and 50wt% porogens (composed of 36wt% of 1,4-butanediol, 54wt% 1-propanol and 10wt% water). Baseline resolution of analytes was achieved through a mobile phase of acetonitrile/water in gradient elution mode. Additionally, dispersive liquid-liquid microextraction (DLLME) was combined with both cLC-UV-Vis and cLC-MS to achieve the determination of parabens in human urine and serum samples with very low limits of detection. Satisfactory intra- and inter-day repeatabilities were obtained in UV-Vis and MS detection, although the latter provided lower detection limits (up to 300-fold) than the UV-Vis detection. Recoveries for the target analytes from spiked biological samples ranged from 95.2% to 106.7%. The proposed methodology for the ultra-low determination of parabens in human urine and serum samples is simple and fast, the consumption of reagents is very low, and very small samples can be analyzed. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. On-line anion exchange solid-phase extraction coupled to liquid chromatography with fluorescence detection to determine quinolones in water and human urine.

    Science.gov (United States)

    Lara, Francisco J; Del Olmo-Iruela, Monsalud; García-Campaña, Ana M

    2013-10-04

    An analytical method based on on-line solid-phase extraction coupled to liquid chromatography with fluorescence detection has been developed to determine quinolones in tap water and human urine. A home-made setup was used to percolate 10 mL of sample through a solid-phase extraction column. Analytes were retained onto the sorbent by an anion exchange mechanism which ensures an optimum compatibility with the subsequent chromatographic separation. A C-18 column containing core-shell particles (2.6 μm) was used to achieve peak efficiencies up to 200,000 plates/m, at a flow rate of 1.2 mL/min and without the need for special pumps. The method allowed the determination of 11 quinolones directly in tap water samples in less than 20 min and with limits of detection ranging between 7 and 110 ng/L. The sensitivity achieved made possible the direct determination of 9 quinolones in human urine without any sample treatment, just dilution with water. Relative recoveries between 94 and 109% were obtained meaning that the matrix effect in human urine is negligible after dilution. Satisfactory results were also obtained in terms of precision since relative standard deviations were always below 13%.

  11. Spectrophotometric and chemometric methods for determination of imipenem, ciprofloxacin hydrochloride, dexamethasone sodium phosphate, paracetamol and cilastatin sodium in human urine

    Science.gov (United States)

    El-Kosasy, A. M.; Abdel-Aziz, Omar; Magdy, N.; El Zahar, N. M.

    2016-03-01

    New accurate, sensitive and selective spectrophotometric and chemometric methods were developed and subsequently validated for determination of Imipenem (IMP), ciprofloxacin hydrochloride (CIPRO), dexamethasone sodium phosphate (DEX), paracetamol (PAR) and cilastatin sodium (CIL) in human urine. These methods include a new derivative ratio method, namely extended derivative ratio (EDR), principal component regression (PCR) and partial least-squares (PLS) methods. A novel EDR method was developed for the determination of these drugs, where each component in the mixture was determined by using a mixture of the other four components as divisor. Peak amplitudes were recorded at 293.0 nm, 284.0 nm, 276.0 nm, 257.0 nm and 221.0 nm within linear concentration ranges 3.00-45.00, 1.00-15.00, 4.00-40.00, 1.50-25.00 and 4.00-50.00 μg mL- 1 for IMP, CIPRO, DEX, PAR and CIL, respectively. PCR and PLS-2 models were established for simultaneous determination of the studied drugs in the range of 3.00-15.00, 1.00-13.00, 4.00-12.00, 1.50-9.50, and 4.00-12.00 μg mL- 1 for IMP, CIPRO, DEX, PAR and CIL, respectively, by using eighteen mixtures as calibration set and seven mixtures as validation set. The suggested methods were validated according to the International Conference of Harmonization (ICH) guidelines and the results revealed that they were accurate, precise and reproducible. The obtained results were statistically compared with those of the published methods and there was no significant difference.

  12. Distribution of rare earth elements in agricultural soil and human body (scalp hair and urine) near smelting and mining areas of Hezhang, China

    Institute of Scientific and Technical Information of China (English)

    玛瑞亚; 季宏兵; 高阳; 丁淮剑; 李彩

    2016-01-01

    Rare earth elements (REEs) in recent decade are widely used and lead to the accumulation of REE in the environment and human body. The aim of this study was to evaluate the concentrations of REEs in soil and human body (scalp hair and urine) of peo-ple living in agricultural soil near smelting and mining areas in Hezhang County, China. The results showed that mean concentrations of determined REEs in agricultural soil from smelting areas were higher than background. However, concentration was slightly higher in soil in mining area. In addition, REEs concentrations of hair and urine in smelting areas were higher than those in mining areas.ΣREEs for soil in mining and smelting areas were 177.79 and 277.06 mg/kg, respectively.ΣREEs for hair in mining and smelting were 1.13 and 1.55 mg/kg, respectively, andΣREEs for urine in mining and smelting were 0.58 and 0.59 µg/L, respectively. Results showed that La, Ce and Nd were enriched in soil, hair and urine. Eu in smelting area showed a positive anomaly. In smelting and mining areas, females were more likely than male to expose to REEs. The relationship between REEs concentration and age group showed that hair’s high concentrations of REE existed in 18–40 years age for people from smelting areas and females from mining areas. While high concentrations distributed in the age of 41–65 for males from mining area. However, urine did not present similar distribution for different age group. Compared with hair and urine, soil showed the same distribution of REEs. And according to the Ce/Ce* value vs. LaN/YbN ratio showed that hair and soil tended to increase, with the stability of Ce/Ce* value. Thus the distri-bution of REEs in soil was closely related with the accumulation in human body. This is a preliminary study which may be suggested to the other research, and this study data may be useful for adding up the data pool on REEs levels in China.

  13. Maduramicin. alpha. : Characterization of sup 14 C-derived residues in turkey excreta

    Energy Technology Data Exchange (ETDEWEB)

    Stout, S.J.; daCunha, A.R.; Lee, A. (American Cyanamid Co., Princeton, NJ (United States)); Jinn Wu; King, K.G. (XenoBiotic Labs., Inc., Princeton, NJ (United States))

    1991-02-01

    Maduramicin {alpha}, a highly potent polyether ionophore antibiotic for preventing coccidiosis in poultry, is passed predominantly in turkey excreta following oral feeding. Following isolation and purification, the turkey excreta metabolites were characterized primarily by liquid chromatography/mass spectrometry and liquid chromatography/tandem mass spectrometry. Maduramicin {alpha} and its metabolites generate a characteristic pair of ions corresponding to (M + NH{sub 4}){sup +} and (M + Na){sup +} which assist in differentiating the metabolites from matrix coextractives. These two ions also fragment differently in tandem mass spectrometry, thus providing structural information for characterizing the nature of unknown metabolites. The primary metabolic pathway of maduramicin {alpha} in the turkey is O-demethylation at one or more of the methoxy groups. Hydroxylation and glucuronide conjugation are minor secondary metabolic processes.

  14. Strong-LAMP: A LAMP Assay for Strongyloides spp. Detection in Stool and Urine Samples. Towards the Diagnosis of Human Strongyloidiasis Starting from a Rodent Model

    Science.gov (United States)

    Gandasegui, Javier; Bajo Santos, Cristina; López-Abán, Julio; Saugar, José María; Rodríguez, Esperanza; Vicente, Belén; Muro, Antonio

    2016-01-01

    Background Strongyloides stercoralis, the chief causative agent of human strongyloidiasis, is a nematode globally distributed but mainly endemic in tropical and subtropical regions. Chronic infection is often clinically asymptomatic but it can result in severe hyperinfection syndrome or disseminated strongyloidiasis in immunocompromised patients. There is a great diversity of techniques used in diagnosing the disease, but definitive diagnosis is accomplished by parasitological examination of stool samples for morphological identification of parasite. Until now, no molecular method has been tested in urine samples as an alternative to stool samples for diagnosing strongyloidiasis. This study aimed to evaluate the use of a new molecular LAMP assay in a well-established Wistar rat experimental infection model using both stool and, for the first time, urine samples. The LAMP assay was also clinically evaluated in patients´ stool samples. Methodology/Principal Findings Stool and urine samples were obtained daily during a 28-day period from rats infected subcutaneously with different infective third-stage larvae doses of S. venezuelensis. The dynamics of parasite infection was determined by daily counting the number of eggs per gram of feces from day 1 to 28 post-infection. A set of primers for LAMP assay based on a DNA partial sequence in the 18S rRNA gene from S. venezuelensis was designed. The set up LAMP assay (namely, Strong-LAMP) allowed the sensitive detection of S. venezuelensis DNA in both stool and urine samples obtained from each infection group of rats and was also effective in S. stercoralis DNA amplification in patients´ stool samples with previously confirmed strongyloidiasis by parasitological and real-time PCR tests. Conclusions/Significance Our Strong-LAMP assay is an useful molecular tool in research of a strongyloidiasis experimental infection model in both stool and urine samples. After further validation, the Strong-LAMP could also be potentially

  15. Monolithic silica spin column extraction and simultaneous derivatization of amphetamines and 3,4-methylenedioxyamphetamines in human urine for gas chromatographic-mass spectrometric detection

    Energy Technology Data Exchange (ETDEWEB)

    Nakamoto, Akihiro [Scientific Investigation Laboratory, Hiroshima Prefectural Police Headquarters, Kohnan 2-26-3, Naka-ku, Hiroshima 730-0825 (Japan); Nishida, Manami [Hiroshima University Technical Center, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8551 (Japan); Saito, Takeshi [Department of Emergency and Critical Care Medicine, Tokai University School of Medicine, Shimokasuya 143, Isehara, Kanagawa 259-1143 (Japan); Kishiyama, Izumi; Miyazaki, Shota [GL Sciences Inc., Sayamagahara 237-2, Iruma, Saitama 358-0032 (Japan); Murakami, Katsunori [Scientific Investigation Laboratory, Hiroshima Prefectural Police Headquarters, Kohnan 2-26-3, Naka-ku, Hiroshima 730-0825 (Japan); Nagao, Masataka [Department of Forensic Medicine, Graduate School of Biomedical Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8551 (Japan); Namura, Akira, E-mail: namera@hiroshima-u.ac.jp [Department of Forensic Medicine, Graduate School of Biomedical Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima 734-8551 (Japan)

    2010-02-19

    A simple, sensitive, and specific method with gas chromatography-mass spectrometry was developed for simultaneous extraction and derivatization of amphetamines (APs) and 3,4-methylenedioxyamphetamines (MDAs) in human urine by using a monolithic silica spin column. All the procedures, such as sample loading, washing, and elution were performed by centrifugation. APs and MDAs in urine were adsorbed on the monolithic silica and derivatized with propyl chloroformate in the column. Methamphetamine-d{sub 5} was used as an internal standard. The linear ranges were 0.01-5.0 {mu}g mL{sup -1} for methamphetamine (MA) and 3,4-methylenedioxymethamphetamine (MDMA) and 0.02-5.0 {mu}g mL{sup -1} for amphetamine (AP) and 3,4-methylenedioxyamphetamine (MDA) (coefficient of correlation {>=}0.995). The recovery of APs and MDAs in urine was 84-94%, and the relative standard deviation of the intra- and interday reproducibility for urine samples containing 0.1, 1.0, and 4.0 {mu}g mL{sup -1} of APs and MDAs ranged from 1.4% to 13.6%. The lowest detection limit (signal-to-noise ratio {>=} 3) in urine was 5 ng mL{sup -1} for MA and MDMA and 10 ng mL{sup -1} for AP and MDA. The proposed method can be used to perform simultaneous extraction and derivatization on spin columns that have been loaded with a small quantity of solvent by using centrifugation.

  16. Simultaneous determination of amitraz, chlordimeform, formetanate and their main metabolites in human urine by high performance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Gao, Xue; Tan, Yanglan; Guo, Hao

    2017-03-11

    A rapid, simple and reliable high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed for simultaneous determination of amitraz, chlordimeform, formetanate and their main metabolites, N-(2,4-dimethylphenyl)-N-methyl-formamidine (DMPF), 2,4-dimethylformamidine (DMF), 2,4-dimethylaniline (DMA), 4-chloro-2-methylaniline and 3-hydroxyacetanilide in human urine. The urine samples were mixed with buffer solutions (pH 8) and subsequently cleaned up by solid supported liquid/liquid extraction (SLE). The target analytes were efficiently separated with a Waters Atlantis T3 column (150mm×4.6mm, 5μm), ionized with electrospray ion source in positive mode, and quantitatively determined by tandem mass spectrometry in the multiple reaction monitoring (MRM) mode. In order to minimize matrix effects, the matrix-matched calibration curves of eight analytes were adopted with correlation coefficients (R(2)) above 0.99. The method were further validated by determining the limits of detection (LODs, 0.3-0.6ng/mL), the limits of quantitation (LOQs, 1.0-2.0ng/mL) and recoveries (89.1%-108.4%) with intra-day and inter-day relative standard deviation (RSD, <11%). The established method was applied and demonstrated in a real case by assaying a urine sample from a female poisoned by formetanate. The achieved results proved this method to be rapid, sensitive and accurate for simultaneous quantitation of eight analytes in human urine for intended forensic cases of human poisoning.

  17. Quantitative analysis of unconjugated and total bisphenol A in human urine using solid-phase extraction and UPLC-MS/MS: method implementation, method qualification and troubleshooting.

    Science.gov (United States)

    Buscher, Brigitte; van de Lagemaat, Dick; Gries, Wolfgang; Beyer, Dieter; Markham, Dan A; Budinsky, Robert A; Dimond, Stephen S; Nath, Rajesh V; Snyder, Stephanie A; Hentges, Steven G

    2015-11-15

    The aim of the presented investigation was to document challenges encountered during implementation and qualification of a method for bisphenol A (BPA) analysis and to develop and discuss precautions taken to avoid and to monitor contamination with BPA during sample handling and analysis. Previously developed and published HPLC-MS/MS methods for the determination of unconjugated BPA (Markham et al. Journal of Analytical Toxicology, 34 (2010) 293-303) [17] and total BPA (Markham et al. Journal of Analytical Toxicology, 38 (2014) 194-203) [20] in human urine were combined and transferred into another laboratory. The initial method for unconjugated BPA was developed and evaluated in two independent laboratories simultaneously. The second method for total BPA was developed and evaluated in one of these laboratories to conserve resources. Accurate analysis of BPA at sub-ppb levels is a challenging task as BPA is a widely used material and is ubiquitous in the environment at trace concentrations. Propensity for contamination of biological samples with BPA is reported in the literature during sample collection, storage, and/or analysis. Contamination by trace levels of BPA is so pervasive that even with extraordinary care, it is difficult to completely exclude the introduction of BPA into biological samples and, consequently, contamination might have an impact on BPA biomonitoring data. The applied UPLC-MS/MS method was calibrated from 0.05 to 25ng/ml. The limit of quantification was 0.1ng/ml for unconjugated BPA and 0.2ng/ml for total BPA, respectively, in human urine. Finally, the method was applied to urine samples derived from 20 volunteers. Overall, BPA can be analyzed in human urine with acceptable recovery and repeatability if sufficient measures are taken to avoid contamination throughout the procedure from sample collection until UPLC-MS/MS analysis.

  18. Metabolism of novel opioid agonists U-47700 and U-49900 using human liver microsomes with confirmation in authentic urine specimens from drug users.

    Science.gov (United States)

    Krotulski, Alex J; Mohr, Amanda L A; Papsun, Donna M; Logan, Barry K

    2017-06-13

    Recently, the number of adverse events, including death, involving novel opioids has continued to increase, providing additional and sustained challenges for forensic and medical communities. Identification of emerging novel opioids can be challenging, compounded by detection windows and unknown metabolic profiles. In this study, human liver microsomes were used for the generation of in vitro metabolic profiles of U-47700 and U-49900. Generated metabolites were analyzed via a SCIEX TripleTOF® 5600+ quadrupole time-of-flight mass spectrometer and resulting data files were processing using MetabolitePilot™. Characterized metabolites were verified in vivo by analysis of authentic human urine specimens collected after analytically confirmed cases of overdose involving U-47700 or U-49900. In total, four metabolites were identified and present in urine specimens for U-47700, and five metabolites for U-49900. N-Desmethyl-U-47700 was determined to be the primary metabolite of U-47700. Parent U-47700 was identified in all urine specimens. N-Desmethyl-U-47700 and N,N-didesmethyl-U-47700 were structurally confirmed for the first time during this study following acquisition of standard reference material. N-Desethyl-U-49900 was determined to be the primary metabolite of U-49900 following microsomal incubations, while N,N-didesethyl-N-desmethyl-U-49900 was the most abundant in a urine specimen. Similarities in metabolic transformation were identified between U-47700 and U-49900, resulting in a common metabolite and isomeric species. These phenomena should be considered in cases involving U-47700 or U-49900. This study is the first to map the metabolic profiles of U-47700 and U-49900 using human liver microsomes, as well as the first to report any literature involving U-49900 and analysis of case specimens. Copyright © 2017 John Wiley & Sons, Ltd.

  19. Cross validation of gas chromatography-flame photometric detection and gas chromatography-mass spectrometry methods for measuring dialkylphosphate metabolites of organophosphate pesticides in human urine.

    Science.gov (United States)

    Prapamontol, Tippawan; Sutan, Kunrunya; Laoyang, Sompong; Hongsibsong, Surat; Lee, Grace; Yano, Yukiko; Hunter, Ronald Elton; Ryan, P Barry; Barr, Dana Boyd; Panuwet, Parinya

    2014-01-01

    We report two analytical methods for the measurement of dialkylphosphate (DAP) metabolites of organophosphate pesticides in human urine. These methods were independently developed/modified and implemented in two separate laboratories and cross validated. The aim was to develop simple, cost effective, and reliable methods that could use available resources and sample matrices in Thailand and the United States. While several methods already exist, we found that direct application of these methods required modification of sample preparation and chromatographic conditions to render accurate, reliable data. The problems encountered with existing methods were attributable to urinary matrix interferences, and differences in the pH of urine samples and reagents used during the extraction and derivatization processes. Thus, we provide information on key parameters that require attention during method modification and execution that affect the ruggedness of the methods. The methods presented here employ gas chromatography (GC) coupled with either flame photometric detection (FPD) or electron impact ionization-mass spectrometry (EI-MS) with isotopic dilution quantification. The limits of detection were reported from 0.10ng/mL urine to 2.5ng/mL urine (for GC-FPD), while the limits of quantification were reported from 0.25ng/mL urine to 2.5ng/mL urine (for GC-MS), for all six common DAP metabolites (i.e., dimethylphosphate, dimethylthiophosphate, dimethyldithiophosphate, diethylphosphate, diethylthiophosphate, and diethyldithiophosphate). Each method showed a relative recovery range of 94-119% (for GC-FPD) and 92-103% (for GC-MS), and relative standard deviations (RSD) of less than 20%. Cross-validation was performed on the same set of urine samples (n=46) collected from pregnant women residing in the agricultural areas of northern Thailand. The results from split sample analysis from both laboratories agreed well for each metabolite, suggesting that each method can produce

  20. Studies on the utilization of ensiled poultry excreta as a ration for replacement stock of cross-bred dairy cattle

    Energy Technology Data Exchange (ETDEWEB)

    Jayal, M.M.; Jain, V.K.; Pathak, N.N.

    1981-04-01

    Four silages, of green maize (silage A) and poultry excreta + green maize (silage B) in experiment (1), poultry excreta + Johnson grass (silage C) in experiment (2), and poultry excreta + green maize (silage D) in experiment (3), were studied. In silages (C) and (D) 8% molasses was added before ensiling. The pH ranged between 4.3 and 4.6 in the different silages. In experiment (1a) green maize silage was fed alone: the poultry excreta + green maize silage was fed along in experiment (1b) and with 1 kg molasses in experiment (1c); the silages were fed with 0.5 kg wheat bran in experiment (2) and with 0.5 kg crushed maize in experiment (3). The voluntary dry matter intake was lowest in experiment (2) and highest in experiment (3). The digestibility of proximate principles, except that of crude protein, improved on supplementation with molasses (1c). Supplementation with maize resulted in increased digestibility of proximate principles except crude fibre. The digestible crude protein content increased with the incorporation of poultry excreta and was proportional to the poultry excreta content of the silages. The balances of nitrogen, calcium and phosphorus were positive. The mean daily gain was 121, 91, 151, 194 and 218 g in experiments (1a), (1b), (1c), (2) and (3) respectively. Inadequate energy intake adversely affected the body weight gain. Such silages, being deficient in energy content, warrant supplementation with higher quantities of energy-rich feeds for supporting production functions.

  1. Metabolic profile of Fructus Gardeniae in human plasma and urine using ultra high-performance liquid chromatography coupled with high-resolution LTQ-orbitrap mass spectrometry.

    Science.gov (United States)

    Wang, Gao-Wa; Bao, Burenbatu; Han, Zhi-Qiang; Han, Qing-Yu; Yang, Xiu-Lan

    2016-10-01

    1. In China, Fructus Gardeniae was used as a traditional Chinese medicine (TCM) with a wide array of biological activities. The bioactive components identified in Fructus Gardeniae mainly included iridoids, flavonids, pigments, and so on. Among them, iridoids were regarded as important compounds in Fructus Gardeniae. Though analyses of the constituents in biological samples after oral administration of Fructus Gardeniae effective fraction or its active compounds have been reported, few efforts have been made to investigate the metabolic profile of Fructus Gardeniae in humans. In this study, the constituents and metabolites of Fructus Gardeniae in human blood and urine after oral administration of Fructus Gardeniae were investigated using ultra high-performance liquid chromatography (UHPLC) coupled with high-resolution LTQ-Orbitrap mass spectrometery. 2. Totally, 14 constituents (two parent compounds and 12 metabolites) of Fructus Gardeniae were identified in human plasma and urine either by comparing the retention time and mass spectrometry data with that of reference compounds or by the accurate high-resolution MS/MS data of the chemicals. The compounds identified were mainly iridoid glycosides such as geniposide and the derivatives of genipin-O-glucuronide. Among them, 11 metabolites were detected in human plasma and urine while the other three metabolites including geniposidic acid (M1), demethylation derivative of genipin-O-glucuronide (M2), and dehydration product of mono-hydroxylated genipin-O-glucuronide (M9) were only discovered in human urine. Further, the possible metabolic pathways of Fructus Gardeniae in vivo were proposed and the peak area-time curve of the most abundant metabolite genipin-O-glucuronide (M13) in human plasma after oral administration of Fructus Gardeniae was depicted. The results suggested that a metabolic difference existed between rats and humans. 3. The results obtained in the present research would provide basic information to

  2. Evaluation of coupling reversed phase, aqueous normal phase, and hydrophilic interaction liquid chromatography with Orbitrap mass spectrometry for metabolomic studies of human urine.

    Science.gov (United States)

    Zhang, Tong; Creek, Darren J; Barrett, Michael P; Blackburn, Gavin; Watson, David G

    2012-02-21

    In this study, we assessed three liquid chromatographic platforms: reversed phase (RP), aqueous normal phase (ANP), and hydrophilic interaction (HILIC) for the analysis of polar metabolite standard mixtures and for their coverage of urinary metabolites. The two zwitterionic HILIC columns showed high-quality chromatographic performance for metabolite standards, improved separation for isomers, and the greatest coverage of polar metabolites in urine. In contrast, on the reversed phase column, most metabolites eluted very rapidly with little or no separation. Using an Exactive Orbitrap mass spectrometer with a HILIC liquid chromatographic platform, approximately 970 metabolite signals with repeatable peak areas (relative standard deviation (RSD) ≤ 25%) could be putatively identified in human urine, by elemental composition assignment within a 3 ppm mass error. The ability of the methodology for the verification of nonmolecular ions, which arise from adduct formation, and the possibility of distinguishing isomers could also be demonstrated. Careful examination of the raw data and the use of masses for predicted metabolites produced an extension of the metabolite list for human urine.

  3. Novel method for simultaneous aqueous in situ derivatization of THC and THC-COOH in human urine samples: validation and application to real samples.

    Science.gov (United States)

    Chericoni, S; Battistini, I; Dugheri, S; Pacenti, M; Giusiani, M

    2011-05-01

    The present work describes the validation of a novel aqueous in situ derivatization procedure with trimethyloxonium tetrafluoroborate (TMO) as methylating agent for the simultaneous, quantitative analysis of Δ(9)-tetrahydrocannabinol (THC) and 11-nor-Δ(9)-tetrahydrocannabinol carboxylic acid (THC-COOH) in human urine. The derivatizing agent is directly added to the urine sample and the methyl-derivatives are then recovered by liquid-liquid extraction procedure. Gas chromatography-mass spectrometry was used to detect the derivatives in selected ion monitoring mode. The limits of detection were 0.7 ng/mL for THC and 0.5 ng/mL for THC-COOH, whereas the limits of quantification were 1.9 and 0.9 ng/mL, respectively. The method has been applied to 60 real samples both positive and negative to immunochemical screening test resulting to be very useful and reliable in routine analysis of THC-COOH in human urine for toxicological and forensic purposes.

  4. Determination of Uric Acid in Human Urine by Ion-exclusion Chromatography with UV Detection Using Pure Water as Mobile Phase

    Institute of Scientific and Technical Information of China (English)

    侯升杰; 杨成对; 王辉; 田中一彦; 丁明玉

    2012-01-01

    A simple, rapid and accurate ion-exclusion chromatographic method coupled with a UV detector for the determination of uric acid in human urine samples has been developed. The separation was carried out on an ion-exclusion column using only pure water as mobile phase. The detection wavelength was 254 nm and urine sample was injected directly without any pretreatment. Furthermore, the retention behavior of uric acid on the ion-exclusion column was researched when pure water and 1 mmol·L-1 HCI were used as mobile phase, respectively. The stability of uric acid was also further investigated within 28 days, In this method, the linear range of the calibration curve for uric acid was 0.25--100 mg·L-1, and the detection limit calculated at S/N=3 was 0.02mg·L-1 The proposed ion-exclusion chromatographic method has been used for the determination of uric acid in human urine.

  5. Molecularly imprinted polymer microspheres prepared by Pickering emulsion polymerization for selective solid-phase extraction of eight bisphenols from human urine samples.

    Science.gov (United States)

    Yang, Jiajia; Li, Yun; Wang, Jincheng; Sun, Xiaoli; Cao, Rong; Sun, Hao; Huang, Chaonan; Chen, Jiping

    2015-05-01

    The bisphenol A (BPA) imprinted polymer microspheres were prepared by simple Pickering emulsion polymerization. Compared to traditional bulk polymerization, both high yields of polymer and good control of particle sizes were achieved. The characterization results of scanning electron microscopy and nitrogen adsorption-desorption measurements showed that the obtained molecularly imprinted polymer microsphere (MIPMS) particles possessed regular spherical shape, narrow diameter distribution (30-60 μm), a specific surface area (S(BET)) of 281.26 m(2) g(-1) and a total pore volume (V(t)) of 0.459 cm(3) g(-1). Good specific adsorption capacity for BPA was obtained in the sorption experiment and good class selectivity for BPA and its seven structural analogs (bisphenol F, bisphenol B, bisphenol E, bisphenol AF, bisphenol S, bisphenol AP and bisphenol Z) was demonstrated by the chromatographic evaluation experiment. The MIPMS as solid-phase extraction (SPE) packing material was then evaluated for extraction and clean-up of these bisphenols (BPs) from human urine samples. An accurate and sensitive analytical method based on the MIPMS-SPE coupled with HPLC-DAD has been successfully established for simultaneous determination of eight BPs from human urine samples with detection limits of 1.2-2.2 ng mL(-1). The recoveries of BPs for urine samples at two spiking levels (100 and 500 ng mL(-1) for each BP) were in the range of 81.3-106.7% with RSD values below 8.3%.

  6. Determination of nerve agent metabolites in human urine by isotope-dilution gas chromatography-tandem mass spectrometry after solid phase supported derivatization.

    Science.gov (United States)

    Lin, Ying; Chen, Jia; Yan, Long; Guo, Lei; Wu, Bidong; Li, Chunzheng; Feng, Jianlin; Liu, Qin; Xie, Jianwei

    2014-08-01

    A simple and sensitive method has been developed and validated for determining ethyl methylphosphonic acid (EMPA), isopropyl methylphosphonic acid (IMPA), isobutyl methylphosphonic acid (iBuMPA), and pinacolyl methylphosphonic acid (PMPA) in human urine using gas chromatography-tandem mass spectrometry (GC-MS/MS) coupled with solid phase derivatization (SPD). These four alkyl methylphosphonic acids (AMPAs) are specific hydrolysis products and biomarkers of exposure to classic organophosphorus (OP) nerve agents VX, sarin, RVX, and soman. The AMPAs in urine samples were directly derivatized with pentafluorobenzyl bromide on a solid support and then extracted by liquid-liquid extraction. The analytes were quantified with isotope-dilution by negative chemical ionization (NCI) GC-MS/MS in a selected reaction monitoring (SRM) mode. This method is highly sensitive, with the limits of detection of 0.02 ng/mL for each compound in a 0.2 mL sample of human urine, and an excellent linearity from 0.1 to 50 ng/mL. It is proven to be very suitable for the qualitative and quantitative analyses of degradation markers of OP nerve agents in biomedical samples.

  7. Relationship Not Found Between Blood and Urine Concentrations and Body Mass Index in Humans With Apparently Adequate Boron Status.

    Science.gov (United States)

    Koc, Fulya; Aysan, Erhan; Hasbahceci, Mustafa; Arpaci, Beyza; Gecer, Salih; Demirci, Selami; Sahin, Fikrettin

    2016-06-01

    The impact of boron on the development of obesity remains controversial in the analysis of experimental and clinical data. The objective of this study was to investigate the relationship between blood and urine boron concentrations and obesity in normal, overweight, obese, and morbidly obese subjects in different age groups. A total of 105 subjects were categorized into 12 groups based on body mass index and three different age levels: as young adult (18 to 34 years old), adult (35 to 54 years old), and older adult (greater than 55 years old). Age, gender, body mass index, and blood and urine boron concentrations were recorded for each subject. There were 50 women and 55 men, with a mean age of 44.63 ± 17.9 years. Blood and urine boron concentrations were similar among the groups (p = 0.510 and p = 0.228, respectively). However, a positive correlation between age and blood boron concentration (p = 0.001) was detected in contrast to the presence of a negative correlation between age and urine boron concentration (p = 0.027). Multiple linear regression analysis showed that there was no significant relationship between gender, age, and quantitative values of body mass index for each subject, and blood and urine boron concentrations. Although the relationship between boron and obesity has not been confirmed, changes of blood and urine boron concentrations with age may have some physiologic sequences to cause obesity.

  8. Proteins of human urine. II. Identification by two-dimensional electrophoresis of a new candidate marker for prostatic cancer

    Energy Technology Data Exchange (ETDEWEB)

    Edwards, J.J. (Argonne National Lab., IL); Anderson, N.G.; Tollaksen, S.L.; von Eschenbach, A.C.; Guevara, J. Jr.

    1982-01-01

    A protein series common to the urine and prostatic tissue of 16 of 17 patients with prostatic adenocarcinoma has been identified by high-resolution two-dimensional gel electrophoresis. These proteins, designated PCA-1, have a relative molecular mass in sodium dodecyl sulfate of about 40,000. Analyses of urines from eight age-matched controls, seven patients with other types of urogenital malignancies, two patients with benign prostatic hyperplasia, and five patients with malignancies not associated with the urogenital system failed to show PCA-1 in the patterns. These preliminary findings suggest that this protein should be systematically investigated as a candidate marker for prostatic adenocarcinoma in man.

  9. Sensitive non-isotopic DNA hybridisation assay or immediate-early antigen detection for rapid identification of human cytomegalovirus in urine.

    Science.gov (United States)

    Kimpton, C P; Morris, D J; Corbitt, G

    1991-04-01

    A sensitive non-radioactive DNA hybridisation assay employing digoxigenin-labelled probes was compared with immediate-early antigen detection and conventional virus isolation for the identification of human cytomegalovirus (HCMV) in 249 urine samples. Of 44 specimens yielding HCMV by virus isolation, more were positive by DNA hybridisation (32; 73%) than by immediate-early antigen detection (25; 52%) (P = 0.05). The specificity of the hybridisation assay in 45 apparently falsely positive specimens was supported by detection of HCMV DNA in 40 of these specimens using the polymerase chain reaction. Many urine specimens may thus contain large amounts of non-viable virus or free viral DNA. Evaluation of various protocols for the extraction and denaturation of virus DNA prior to hybridisation showed that proteinase K digestion with phenol/chloroform extraction was the most sensitive and reliable procedure. We conclude that the non-radioactive DNA hybridisation assay described is a potentially valuable routine diagnostic test.

  10. Environmental survey 1998. Vol. 3: Human biomonitoring. Concentrations of substances in blood and urine of the German population; Umwelt-Survey 1998. Bd. 3: Human-Biomonitoring. Stoffgehalte in Blut und Urin der Bevoelkerung in Deutschland

    Energy Technology Data Exchange (ETDEWEB)

    Becker, K.; Kaus, S.; Krause, C.; Lepom, P.; Schulz, C.; Seiwert, M.; Seifert, B.

    2002-03-01

    The occurrence of precious metals in the environment is increasing, due primarily to their use in catalytic converters for motor vehicles, and they were therefore integrated into the study programme in 1998. Urine samples of 1080 subjects were analysed for gold, platinum and iridium, resulting in mean concentrations of 46 ng/l, 2.2 ng/l and 0.2 ng/l. The number of dental inlays, crowns and bridge elements has a clear influence on the mean levels of gold and platinum in urine. Road traffic was not found to be a signficant factor. Nicotine and cotinine are common markers for exposure to tobacco smoke. Non-smokers exhibit a mean nicotine level in urine of less than 2 {mu}g/l and a cotinine level of less than 4 {mu}gl. Nicotine and cotinine levels in urine increase as a function of the number of cigarettes smoked per day, reaching values of 1080 {mu}g/l and 2060 {mu}g/l, respectively, when consumption exceeds 20 cigarettes per day. Exposure to environmental tobacco smoke is also reflected by nicotine and cotinine levels in urine. The present report provides representative data for environmentally related health monitoring and reporting at national level. The data serve as the basis for the formulation of reference values characterising the population's internal exposure to environmental contaminants. The report shows trends over time and regional differences in pollution, and identifies exposure pathways. The results of the Environmental Survey serve inter alia to achieve nationally harmonised methodologies for assessment, health-related environmental reporting, and the development and monitoring of prevention, intervention and abatement strategies within the scope of health and environmental policy actions. (orig.)

  11. Ultrasound-assisted hydrolysis of conjugated parabens in human urine and their determination by UPLC-MS/MS and UPLC-HRMS.

    Science.gov (United States)

    Schlittenbauer, Linda; Seiwert, Bettina; Reemtsma, Thorsten

    2016-02-01

    Parabens are preservatives widely used in personal care products, pharmaceutical formulations as well as in food, and they are considered endocrine disruptors. For application in biomonitoring studies we developed a method for the determination of eight parabens from human urine. Sample preparation was enhanced and simplified by the combination of ultrasound-assisted enzymatic hydrolysis of conjugates (glucuronide and sulfate) followed by an extraction-free cleanup step. Quantification, using deuterated parabens as internal standards, was performed by ultrahigh-performance liquid chromatography coupled to either triple-quadrupole (UPLC-QqQ) or time-of-flight (UPLC-QqTOF) mass spectrometry. Full chromatographic separation of three butyl paraben isomers was achieved. Limits of quantification for both mass analyzers ranged from 0.1 to 0.5 μg/L for methyl, ethyl, n-/isopropyl, n-/isobutyl, and benzyl paraben in 200 μL of urine sample. The method was tested for applicability and showed high precision (intra- and interday 0.9-14.5%) as well as high accuracy (relative recovery 95-132%). A total of 39 urine samples were analyzed by both mass analyzers. The results agreed well, with a trend to higher deviation at low concentrations (less than 10 μg/L). Methyl, ethyl, and n-propyl paraben were detected most frequently (in more than 87% of the samples) with median concentrations ranging from 0.8 to 16.6 μg/L. Female urine showed higher median concentrations for all parabens, which may indicate higher exposure due to lifestyle. This method permits accurate and high-throughput analysis of parabens for epidemiological studies. Further, the UPLC-QqTOF approach provides additional information on human exposure to other compounds by post-acquisition analysis.

  12. Simultaneous determination of ivabradine and N-desmethylivabradine in human plasma and urine using a LC-MS/MS method: application to a pharmacokinetic study

    Directory of Open Access Journals (Sweden)

    Chengtao Lu

    2012-04-01

    Full Text Available A sensitive and specific liquid-chromatography tandem mass spectrometry (LC-MS/MS assay has been developed and validated for the simultaneous quantification of ivabradine and its active metabolite N-desmethylivabradine in human plasma and urine. The assay employed a single liquid–liquid extraction of the analytes from plasma and urine samples, and diazepam was used as internal standard (IS. The chromatographic separation was achieved on a Diamonsil C18 column (150 mm×4.6 mm, 5 μm, Dikma using a mixture of methanol and aqueous 5 mM ammonium acetate buffer containing 0.2% formic acid (80:20, v/v as mobile phase. The assay for ivabradine and N-desmethylivabradine in plasma showed good linearity (r≥0.99 over the ranges 0.1013–101.3 ng/mL and 0.085–25.5 ng/mL, respectively. The assay for ivabradine and N-desmethylivabradine in urine showed good linearity (r≥0.99 over the ranges 10.13–6078 ng/mL and 8.5–850 ng/mL, respectively. The intra- and inter-day accuracy and precision values were found to be within the assay variability limits (RSD<15% in accordance with FDA guidelines. The methods were successfully used for evaluating the pharmacokinetic properties of ivabradine and N-desmethylivabradine in human plasma and urine in Chinese healthy volunteers.

  13. Diagnostic accuracy and applicability of a PCR system for the detection of Schistosoma mansoni DNA in human urine samples from an endemic area.

    Directory of Open Access Journals (Sweden)

    Martin Johannes Enk

    Full Text Available Schistosomiasis caused by Schistosoma mansoni, one of the most neglected human parasitoses in Latin America and Africa, is routinely confirmed by microscopic visualization of eggs in stool. The main limitation of this diagnostic approach is its lack of sensitivity in detecting individual low worm burdens and consequently data on infection rates in low transmission settings are little reliable. According to the scientific literature, PCR assays are characterized by high sensitivity and specificity in detecting parasite DNA in biological samples. A simple and cost effective extraction method for DNA of Schistosoma mansoni from urine samples in combination with a conventional PCR assay was developed and applied in an endemic area. This urine based PCR system was tested for diagnostic accuracy among a population of a small village in an endemic area, comparing it to a reference test composed of three different parasitological techniques. The diagnostic parameters revealed a sensitivity of 100%, a specificity of 91.20%, positive and negative predictive values of 86.25% and 100%, respectively, and a test accuracy of 94.33%. Further statistical analysis showed a k index of 0.8806, indicating an excellent agreement between the reference test and the PCR system. Data obtained from the mouse model indicate the infection can be detected one week after cercariae penetration, opening a new perspective for early detection and patient management during this stage of the disease. The data indicate that this innovative PCR system provides a simple to handle and robust diagnostic tool for the detection of S. mansoni DNA from urine samples and a promising approach to overcome the diagnostic obstacles in low transmission settings. Furthermore the principals of this molecular technique, based on the examination of human urine samples may be useful for the diagnosis of other neglected tropical diseases that can be detected by trans-renal DNA.

  14. Metabolism studies of the Kratom alkaloids mitraciliatine and isopaynantheine, diastereomers of the main alkaloids mitragynine and paynantheine, in rat and human urine using liquid chromatography-linear ion trap-mass spectrometry.

    Science.gov (United States)

    Philipp, Anika A; Wissenbach, Dirk K; Weber, Armin A; Zapp, Josef; Maurer, Hans H

    2011-05-01

    Mitragyna speciosa (Kratom in Thai), native in Southeast Asia, is increasingly misused as a herbal drug of abuse. During metabolism studies on the Kratom alkaloids mitragynine, its diastereomers speciogynine and speciociliatine as well as paynantheine in rats and humans, further isomeric compounds were detected in Kratom users' urine. The question arose whether these compounds were formed from the low abundant, isomeric alkaloids mitraciliatine (MC) and isopaynantheine (ISO-PAY). Therefore, the aim of the presented study was to identify using liquid chromatography-linear ion trap-mass spectrometry their phase I and II metabolites in rat urine after administration of pure MC or ISO-PAY, to confirm their formation in humans, and finally to confirm whether the above-mentioned isomeric compounds in human urine represent MC and ISO-PAY and/or their metabolites. The metabolic pathways of both alkaloids in rats were found to be comparable to those of their corresponding diastereomers. In the human urines tested, not all metabolites found in rats could be detected because of the much lower amounts of MC and ISO-PAY in Kratom. However, all the above-mentioned so far unknown isomeric compounds could be identified in the human urine samples as MC, ISO-PAY and/or their metabolites. The used LC separation was also suitable for the differentiation of all other Kratom alkaloids and their metabolites in human urine.

  15. Reuse--the ultimate sink? Urine-diverting toilets to protect groundwater quality and fertilise urban agriculture.

    Science.gov (United States)

    Drangert, J O

    2000-01-01

    People are concerned about water and food scarcity and the threats that faecal pollution and malnutrition pose to their health. Ecological sanitation systems open up for new, constructive options in sanitation, not least in poor periurban areas. The purpose of developing a no-mix excreta disposal system is to save water, to reduce wastewater treatment problems, and to protect groundwater quality as well as to recirculate nutrients from urine. In this paper all these aspects will be dealt with comprehensively.

  16. FIRST REPORT ON Cryptococcus neoformans IN PIGEON EXCRETA FROM PUBLIC AND RESIDENTIAL LOCATIONS IN THE METROPOLITAN AREA OF CUIABÁ, STATE OF MATO GROSSO, BRAZIL

    Directory of Open Access Journals (Sweden)

    Doracilde Terumi Takahara

    2013-12-01

    Full Text Available SUMMARY Cryptococcosis is a severe systemic mycosis caused by two species of Cryptococcus that affect humans and animals: C. neoformans and C. gattii. Cosmopolitan and emergent, the mycosis results from the interaction between a susceptible host and the environment. The occurrence of C. neoformans was evaluated in 122 samples of dried pigeon excreta collected in 49 locations in the City of Cuiabá, State of Mato Grosso, Brazil, including public squares (n = 5, churches (n = 4, educational institutions (n = 3, health units (n = 8, open areas covered with asbestos (n = 4, residences (n = 23, factory (n = 1 and a prison (n = 1. Samples collected from July to December of 2010 were seeded on Niger seed agar (NSA. Dark brown colonies were identified by urease test, carbon source assimilation tests and canavanine-glycine-bromothymol blue medium. Polymerase chain reaction primer pairs specific for C. neoformans were also used for identification. Cryptococcus neoformans associated to pigeon excreta was isolated from eight (6.6% samples corresponding to six (12.2% locations. Cryptococcus neoformans was isolated from urban areas, predominantly in residences, constituting a risk of acquiring the disease by immunocompromised and immunocompetent individuals.

  17. Quasi-targeted analysis of hydroxylation-related metabolites of polycyclic aromatic hydrocarbons in human urine by liquid chromatography-mass spectrometry.

    Science.gov (United States)

    Tang, Caiming; Tan, Jianhua; Fan, Ruifang; Zhao, Bo; Tang, Caixing; Ou, Weihui; Jin, Jiabin; Peng, Xianzhi

    2016-08-26

    Metabolite identification is crucial for revealing metabolic pathways and comprehensive potential toxicities of polycyclic aromatic hydrocarbons (PAHs) in human body. In this work, a quasi-targeted analysis strategy was proposed for metabolite identification of monohydroxylated polycyclic aromatic hydrocarbons (OH-PAHs) in human urine using liquid chromatography triple quadruple mass spectrometry (LC-QqQ-MS/MS) combined with liquid chromatography high resolution mass spectrometry (LC-HRMS). Potential metabolites of OH-PAHs were preliminarily screened out by LC-QqQ-MS/MS in association with filtering in a self-constructed information list of possible metabolites, followed by further identification and confirmation with LC-HRMS. The developed method can provide more reliable and systematic results compared with traditional untargeted analysis using LC-HRMS. In addition, data processing for LC-HRMS analysis were greatly simplified. This quasi-targeted analysis method was successfully applied to identifying phase I and phase II metabolites of OH-PAHs in human urine. Five metabolites of hydroxynaphthalene, seven of hydroxyfluorene, four of hydroxyphenanthrene, and three of hydroxypyrene were tentatively identified. Metabolic pathways of PAHs in human body were putatively revealed based on the identified metabolites. The experimental results will be valuable for investigating the metabolic processes of PAHs in human body, and the quasi-targeted analysis strategy can be expanded to the metabolite identification and profiling of other compounds in vivo.

  18. Routine analysis of amphetamine class drugs as their naphthaquinone derivatives in human urine by high-performance liquid chromatography.

    Science.gov (United States)

    Talwar, D; Watson, I D; Stewart, M J

    1999-12-10

    We describe a simple HPLC method which is suitable for the routine confirmation of immunoassay positive amphetamine urine samples. The precolumn derivisation method employing sodium naphthaquinone-4-sulphonate was found to have adequate sensitivity, selectivity and precision for the measurement of amphetamine, methamphetamine, 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), and 3,4-methylenedioxyethylamphetamine (MDEA) at 500 microg/l cutoff level for confirmatory analysis of amphetamines in urine. The specificity of the method is enhanced by detecting the peaks at two different wavelengths. The ratios of the peak heights measured at the two wavelengths were different for each of the 5 amphetamines analysed. There was no interference from other phenylethylamine analogues that are commonly found in "over the counter" preparations. The HPLC method is compared to a commercial TLC system for detecting amphetamines in urine of drug abusers attending drug rehabilitation programmes. The HPLC confirmatory method described is a viable alternative to GC or to the more complex and costly GC-MS techniques for confirming amphetamine, methamphetamine, MDMA, MDA and MDEA in urine of drug abusers especially when used in a clinical care setting.

  19. A Laboratory Experiment in Pharmaceutical Analysis: Determination of Drugs of Abuse in Human Urine by Thin-Layer Chromatography.

    Science.gov (United States)

    Bailey, Leonard C.

    1979-01-01

    An experiment is described that was developed for a course in Inorganic and Analytical Pharmaceutical Chemistry at Rutgers University to provide pharmacy students with practical experience in the thin-layer chromatography used for the analysis of urine to monitor patient compliance with drug abuse treatment programs. (JMD)

  20. Dynamic versus static ultrasonic sample treatment for the solid-liquid pre-concentration of mercury from human urine.

    Science.gov (United States)

    Patricio, A; Fernandez, C; Mota, A M; Capelo, J L

    2006-05-15

    Dynamic and static ultrasonic procedures involving ultrasonic bath and tandem focused ultrasound (i.e. two probes were used in the same sample treatment) have been assessed in order to implement a reliable solid-liquid back extraction of mercury from commercial resins (dowex and chelex-100), previously used to concentrate Hg(II) from treated urine. The urine had been previously treated with an advanced oxidation process provided by the conjunction of potassium permanganate, hydrochloric acid and high intensity focused ultrasound, which allowed that organic matter degradation was achieved in less than 3min. 95+/-10% of mercury in the certified urine and 97+/-6% of the spiked methyl-mercury was recovered with the dowex resin plus the static ultrasonic procedure, whilst 96+/-11% of the spiked mercury was recovered with the dowex resin plus the dynamic procedure, for which ultrasonication was not necessary. The Hg pre-concentration factor used in this work was 8 (20mL of urine to 2.5mL of acid), but different volume ratios can be used in order to increase this factor.

  1. Determination of selenite and selenate in human urine by ion chromatography and inductively coupled plasma mass spectrometry

    DEFF Research Database (Denmark)

    Gammelgaard, Bente; Jons, O.

    2000-01-01

    monitored in the inductively coupled plasma mass spectrometry (ICP-MS) detector. When the chromatographic system was applied to analysis of urine samples diluted 1 + 1, the selenomethionine signal appeared in the front together with other unresolved selenium species, while the selenite and selenate signals...

  2. Abuse of nutmeg (Myristica fragrans Houtt.): studies on the metabolism and the toxicologic detection of its ingredients elemicin, myristicin, and safrole in rat and human urine using gas chromatography/mass spectrometry.

    Science.gov (United States)

    Beyer, Jochen; Ehlers, Dorothea; Maurer, Hans H

    2006-08-01

    Seeds of nutmeg are used as spice, but they are also abused because of psychotropic effects described after ingestion of large doses. It was postulated that these effects could be attributable to metabolic formation of amphetamine derivatives from the main nutmeg ingredients elemicin (EL), myristicin (MY), and safrole (SA). In a case of a suspected nutmeg abuse, neither such amphetamine derivatives nor the main nutmeg ingredients could be detected in urine. The metabolites of EL, MY, and SA were identified using gas chromatography-mass spectrometry in rat urine and their presence in human urine of the nutmeg abuser was confirmed. The identified metabolites indicated that EL, MY, and SA were once and twice hydroxylated at the side chain. In addition, EL was O-demethylated at 2 positions followed by side chain hydroxylation. MY and SA were demethylenated and subsequently methylated. In the human urine sample, the following metabolites could be identified: O-demethyl elemicin, O-demethyl dihydroxy elemicin, demethylenyl myristicin, dihydroxy myristicin, and demethylenyl safrole. As in the human urine sample, neither amphetamine derivatives nor the main nutmeg ingredients could be detected in the rat urine samples. Finally, toxicologic detection of nutmeg abuse was possible by identification of the described metabolites of the EL, MY, and SA in urine applying the authors' systematic toxicologic analysis procedure using full-scan gas chromatography-mass spectrometry after acid hydrolysis, liquid-liquid extraction of analytes, and microwave-assisted acetylation of extracted analytes.

  3. Determination of mepitiostane metabolites in human urine by liquid chromatography/tandem mass spectrometry for sports drug testing.

    Science.gov (United States)

    Okano, Masato; Sato, Mitsuhiko; Kojima, Asami; Kageyama, Shinji

    2015-11-10

    Mepitiostane (2α,3α-epithio-17β-(1-methoxycyclopentyloxy)-5α-androstane), which is a prodrug of epitiostanol (2α,3α-epitio-5α-androstane-17β-ol), is an epitiosteroid having anti-estrogenic and weak androgenic anabolic activities. The World Anti-Doping Agency prohibits the misuse of mepitiostane by athletes. Detection of the urinary metabolites epitiostanol sulfoxide and epitiostanol was studied using liquid chromatography/mass spectrometry (LC-MS) for doping control purposes. The use of LC-MS provided advantages over gas chromatography/mass spectrometry for detecting heat labile steroids because epitiostanol and epitiostanol sulfoxide were primarily pyrolized to 5α-androst-2-en-17β-ol. The method consists of enzymatic hydrolysis using β-glucuronidase (Escherichia coli), liquid-liquid extraction, and subsequent ultra-performance liquid chromatography/electrospray-tandem mass spectrometry. Epitiostanol sulfoxide was determined at urinary concentrations of 0.5-50ng/mL, recovery was 76.2-96.9%, and assay precision was calculated as 0.9-1.7% (intra-day) and 2.0-6.6% (inter-day). Epitiostanol was determined at urinary concentrations of 0.5-50ng/mL, recovery was 26.1-35.6% and assay precision was calculated as 4.1-4.6% (intra-day) and 3.3-8.5% (inter-day). The limits of detection for epitiostanol sulfoxide and epitiostanol were 0.05ng/mL and 0.10ng/mL, respectively. Epitiostanol sulfoxide and epitiostanol, as their gluco-conjugates, were identified in human urine after oral administration of 10mg mepitiostane. Epitiostanol sulfoxide and epitiostanol could be detected up to 48h and 24h after administration, respectively. The results showed that the detection window of epitiostanol is much shorter than that of epitiostanol sulfoxide. The LC-MS detection of urinary epitiostanol sulfoxide, a specific metabolite with a sulphur atom in its molecular structure, is likely to be able to identify the abuse of mepitiostane.

  4. Normalization to specific gravity prior to analysis improves information recovery from high resolution mass spectrometry metabolomic profiles of human urine.

    Science.gov (United States)

    Edmands, William M B; Ferrari, Pietro; Scalbert, Augustin

    2014-11-04

    Extraction of meaningful biological information from urinary metabolomic profiles obtained by liquid-chromatography coupled to mass spectrometry (MS) necessitates the control of unwanted sources of variability associated with large differences in urine sample concentrations. Different methods of normalization either before analysis (preacquisition normalization) through dilution of urine samples to the lowest specific gravity measured by refractometry, or after analysis (postacquisition normalization) to urine volume, specific gravity and median fold change are compared for their capacity to recover lead metabolites for a potential future use as dietary biomarkers. Twenty-four urine samples of 19 subjects from the European Prospective Investigation into Cancer and nutrition (EPIC) cohort were selected based on their high and low/nonconsumption of six polyphenol-rich foods as assessed with a 24 h dietary recall. MS features selected on the basis of minimum discriminant selection criteria were related to each dietary item by means of orthogonal partial least-squares discriminant analysis models. Normalization methods ranked in the following decreasing order when comparing the number of total discriminant MS features recovered to that obtained in the absence of normalization: preacquisition normalization to specific gravity (4.2-fold), postacquisition normalization to specific gravity (2.3-fold), postacquisition median fold change normalization (1.8-fold increase), postacquisition normalization to urinary volume (0.79-fold). A preventative preacquisition normalization based on urine specific gravity was found to be superior to all curative postacquisition normalization methods tested for discovery of MS features discriminant of dietary intake in these urinary metabolomic datasets.

  5. Detection of Delta9-tetrahydrocannabinolic acid A in human urine and blood serum by LC-MS/MS.

    Science.gov (United States)

    Jung, Julia; Kempf, Juergen; Mahler, Hellmut; Weinmann, Wolfgang

    2007-03-01

    Delta9-tetrahydrocannabinolic acid A (Delta9-THCA-A) is the precursor of Delta9-tetrahydrocannabinol (Delta9-THC) in hemp plants. During smoking, the non-psychoactive Delta9-THCA-A is converted to Delta9-THC, the main psychoactive component of marihuana and hashish. Although the decarboxylation of Delta9-THCA-A to Delta9-THC was assumed to be complete--which means that no Delta9-THCA-A should be detectable in urine and blood serum of cannabis consumers--we found Delta9-THCA-A in the urine and blood serum samples collected from police controls of drivers suspected for driving under the influence of drugs (DUID). For LC-MS/MS analysis, urine and blood serum samples were prepared by solid-phase extraction. Analysis was performed with a phenylhexyl column using gradient elution with acetonitrile. For detection of Delta9-THCA-A, the mass spectrometer (MS) (SCIEX API 365 triple-quadrupole MS with TurboIonSpray source) was operated in the multiple reaction monitoring (MRM) mode using the following transitions: m/z357 --> 313, m/z357 --> 245 and m/z357 --> 191. Delta9-THCA-A could be detected in the urine and blood serum samples of several cannabis consumers in concentrations of up to 10.8 ng/ml in urine and 14.8 ng/ml in serum. The concentration of Delta9-THCA-A was below the Delta9-THC concentration in most serum samples, resulting in molar ratios of Delta9-THCA-A/Delta9-THC of approximately 5.0-18.6%. Only in one case, where a short elapsed time between the last intake and blood sampling is assumed, the molar ratio was 18.6% in the serum. This indicates differences in elimination kinetics, which need to be investigated in detail.

  6. Application of liquid chromatography-accelerator mass spectrometry (LC-AMS) to evaluate the metabolic profiles of a drug candidate in human urine and plasma.

    Science.gov (United States)

    Prakash, Chandra; Shaffer, Christopher L; Tremaine, Larry M; Liberman, Rosa G; Skipper, Paul L; Flarakos, Jimmy; Tannenbaum, Steven R

    2007-08-01

    Metabolite profiling of 100- and 1,000-fold diluted urine and plasma samples from a conventional radiolabeled human ADME study is described using a highly sensitive LC-AMS technique. The concentration of radioactivity and the metabolic profiles in urine and plasma determined using this technique were similar to those employing standard off-line (i.e. LSC) or in-line (i.e. beta-RAM or LC-ARC dynamic-flow) radioactivity monitoring techniques. The results indicate that at a simulated ca. 100 nCi clinical dose, plasma and urine concentrations of (14)C, as well as their metabolic profiles, may be determined routinely by LC-AMS. This approach opens the possibility of using LC-AMS for both the high-throughput quantitation of biological samples and the generation of high-resolution chromatographic profiles of complex mixtures at a lower cost than current AMS analyses that require the conversion of sample carbon to graphite, a laborious and time consuming process.

  7. Occurrence and Profile Characteristics of the Pesticide Imidacloprid, Preservative Parabens, and Their Metabolites in Human Urine from Rural and Urban China.

    Science.gov (United States)

    Wang, Lei; Liu, Tianzhen; Liu, Fang; Zhang, Junjie; Wu, Yinghong; Sun, Hongwen

    2015-12-15

    Knowledge of human exposure to imidacloprid, the most extensively used insecticide, and para-hydroxybenzoic acid esters (parabens), the most extensively used preservative, is insufficient. In this study, 295 urine samples collected from subjects in rural and urban areas in China were analyzed for imidacloprid and four parabens (namely, methyl paraben, ethyl paraben, propyl paraben, and butyl paraben) as well as their major metabolites (namely, 6-chloronicotinic acid (6-ClNA) and para-hydroxybenzoic acid (p-HB)). Imidacloprid was detected in 100% of the urine samples from rural Chinese subjects and 95% of the urine samples from urban Chinese subjects. Concentrations of urinary imidacloprid detected in rural Chinese subjects (geometric mean (GM) = 0.18 ng/mL) were slightly higher than those detected in urban Chinese subjects (GM = 0.15 ng/mL) when the effect of pesticide spraying was excluded. However, concentrations of urinary imidacloprid detected in rural adults increased significantly in the subsequent days of pesticide spraying (GM = 0.62 ng/mL), which could return to the normal levels within 3 days. In contrast, concentrations of urinary parabens detected in rural Chinese subjects (GM = 6.90 ng/mL) were lower than that in urban Chinese subjects (GM = 30.5 ng/mL). In addition, the metabolism characteristics of imidacloprid to 6-ClNA and parabens to p-HB were discussed preliminarily.

  8. Excretion of 3-Mercaptolactate-Cysteine Disulfide, Sulfate and Taurine in human Urine before and after Oral Administration of Sulfur-containing Amino Acids.

    Directory of Open Access Journals (Sweden)

    Yuasa,Shigeki

    1990-06-01

    Full Text Available The excretion of 3-mercaptolactate-cysteine mixed disulfide [S-(2-hydroxy-2-carboxyethylthio-L-cysteine, HCETC], sulfate and taurine in the urine of normal adults was investigated before and after oral administration of L-cysteine and related sulfur-containing amino acids. Before the loading of amino acids, the excretion (mean +/- SD per kg of body weight per day of HCETC, free sulfate and taurine was 0.096 +/- 0.042, 305.7 +/- 66.1 and 31.9 +/- 8.7 mumols, respectively. After the loading of L-cysteine (800 mumols/kg of body weight, the average excretion in the 24-h urine of HCETC increased 2-fold and that of taurine increased 1.6-fold. The average excretion of free sulfate after the L-cysteine loading was 989.4 +/- 145.1 and 388.8 +/- 51.6 mumols/kg per day in the first and second 24-h urine, respectively, indicating that the sulfur corresponding to 85% of the L-cysteine loaded was excreted as free sulfate in 24 h. Administration of L-cystine (400 mumols/kg resulted in similar results. The increase in HCETC after L-cysteine or L-cystine administration indicates that L-cysteine is metabolized in part through the transamination pathway (3-mercaptopyruvate pathway and that an equilibrium exists between the intake and excretion of sulfur in humans.

  9. CM Affi-Gel Blue chromatography of human urine: a simple one-step procedure for obtaining erythropoietin suitable for in vitro erythropoietic progenitor assays.

    Science.gov (United States)

    Krystal, G; Eaves, C J; Eaves, A C

    1984-11-01

    A method for both concentrating and purifying human urinary erythropoietin (Ep) using CM Affi-Gel Blue is described. We have found that up to 40 litres of urine can be processed on a 1 litre gel bed of this material. This gives a 25-50-fold purification of Ep with an apparent Ep recovery in excess of 100%. The high recovery of Ep is probably due, in part, to the removal of inhibitors present in the initial urine. By selecting urine that contains high levels of Ep (greater than 0.5 units/ml), it is possible with this method routinely to obtain preparations with specific activities of 100-300 units of Ep per mg protein. Such preparations are noninhibitory when assayed in either short-term suspension cultures or in longer-term methylcellulose cultures at concentrations up to 5-10 units/ml. Similar tests with these same bioassay systems have shown that other non-Ep stimulating factors (i.e. erythroblast enhancing factor (EEF), granulocyte/macrophage colony stimulating factor (GM-CSF) and burst promoting activity (BPA) ) are also not present at detectable levels. In this study we also show that the loss of biological activity which often occurs when partially purified Ep preparations are stored in solution is markedly reduced in the presence of either 1% bovine serum albumin or 0.1% sodium dodecyl sulphate.

  10. LC-MS/MS quantification of sulforaphane and indole-3-carbinol metabolites in human plasma and urine after dietary intake of selenium-fortified broccoli.

    Science.gov (United States)

    Hauder, Johanna; Winkler, Stefanie; Bub, Achim; Rüfer, Corinna E; Pignitter, Marc; Somoza, Veronika

    2011-08-10

    This study aimed at developing a sensitive LC-MS/MS method for the quantification of sulforaphane (SFN) and indole-3-carbinol metabolites in plasma and urine after dietary intake of regular and selenium-fertilized broccoli using stable isotope dilution analysis. In a three-armed, placebo-controlled, randomized human intervention study with 76 healthy volunteers, 200 g of regular (485 μg of total glucosinolates and <0.01 μg of selenium per gram fresh weight) or selenium-fertilized broccoli (589 μg of total glucosinolates and 0.25 μg of selenium per gram fresh weight) was administered daily for 4 weeks. Glucoraphanin and glucobrassicin metabolites quantified in plasma and urine were SFN-glutathione, SFN-cysteine, SFN-cysteinylglycine, SFN-acetylcysteine, and indole-3-carboxaldehyde, indole-3-carboxylic acid, and ascorbigen, respectively. Dietary intake of selenium-fertilized broccoli increased serum selenium concentration analyzed by means of atomic absorption spectroscopy by up to 25% (p < 0.001), but affected neither glucosinolate concentrations in broccoli nor their metabolite concentrations in plasma and urine compared to regular broccoli.

  11. Development of a cloud point extraction and spectrophotometry-based microplate method for the determination of nitrite in human urine and blood

    Science.gov (United States)

    Zhao, Jiao; Lu, Yunhui; Fan, Chongyang; Wang, Jun; Yang, Yaling

    2015-02-01

    A novel and simple method for the sensitive determination of trace amounts of nitrite in human urine and blood has been developed by combination of cloud point extraction (CPE) and microplate assay. The method is based on the Griess reaction and the reaction product is extracted into nonionic surfactant Triton-X114 using CPE technique. In this study, decolorization treatment of urine and blood was applied to overcome the interference of matrix and enhance the sensitivity of nitrite detection. Multi-sample can be simultaneously detected thanks to a 96-well microplate technique. The effects of different operating parameters such as type of decolorizing agent, concentration of surfactant (Triton X-114), addition of (NH4)2SO4, extraction temperature and time, interfering elements were studied and optimum conditions were obtained. Under the optimum conditions, a linear calibration graph was obtained in the range of 10-400 ng mL-1 of nitrite with limit of detection (LOD) of 2.5 ng mL-1. The relative standard deviation (RSD) for determination of 100 ng mL-1 of nitrite was 2.80%. The proposed method was successfully applied for the determination of nitrite in the urine and blood samples with recoveries of 92.6-101.2%.

  12. Determination of metoprolol and its two metabolites in human plasma and urine by high performance liquid chromatography with fluorescence detection and its application in pharmacokinetics.

    Science.gov (United States)

    Xu, Tao; Bao, Shihui; Geng, Peiwu; Luo, Jun; Yu, Lei; Pan, Peipei; Chen, Yi; Hu, Guoxin

    2013-10-15

    A simple, specific and sensitive HPLC method has been developed for the determination of metoprolol and its two metabolites in human plasma and urine. Separation of metoprolol, α-hydroxymetoprolol, O-desmethylmetoprolol and esmolol (internal standard) was achieved on an Agilent XDB-C18 column (150mm×4.6mm, 5μm) using fluorescence detection with Ex 216nm and Em 312nm. The mobile phase consisted of ACN-H2O-0.1%TFA. The analysis was performed in less than 16min with a flow rate of 0.8mL/min. The assay was linear over the concentration range of 5-600ng/mL and 2.5-300ng/mL for metoprolol and its metabolites, respectively. The LOQ were 5.0 and 2.5ng/mL for plasma and urine, respectively. Good precision and accuracy for metoprolol and its two metabolites were obtained. The extraction recoveries were found to be more than 86.91% both in plasma and urine. At the same time, the method was successfully applied to nine healthy volunteers who had been given an oral tablet of 100mg metoprolol.

  13. Enantioselective determination of metoprolol and its metabolites in human urine high-performance liquid chromatography with fluorescence detection (HPLC-FLD) and tandem mass spectrometry (MS/MS).

    Science.gov (United States)

    Baranowska, Irena; Adolf, Weronika; Magiera, Sylwia

    2015-11-01

    A sensitive, stereoselective assay using solid phase extraction and high-performance liquid chromatography (HPLC) with fluorescence detection (FLD) was developed and validated for the analysis of enantiomers of metoprolol and its metabolites (α-hydroxymetoprolol, O-desmethylmetoprolol). Chiral separation was achieved using a CHIRALCEL OD-RH column, packed with cellulose tris-(3,5-dimethylphenyl-carbamate) stationary phase, employing a mobile phase composed by a mixture of 0.2% diethylamine in water and acetonitrile in gradient elution mode. Linear calibration curves were obtained over the range of 0.025-2.0μg/mL (R(2)>0.994) in urine for both enantiomers of metoprolol and its metabolites with quantitation limit of 0.025μg/mL. Intra and inter-day precision and accuracy were below 15% for both metoprolol and metabolites enantiomers. The recovery of enantiomer of metoprolol and its metabolite was greater than 68.0%, utilizing a SPE procedure. The method was tested with urine quality control samples and human urine fractions after administration of 50mg rac-metoprolol.

  14. Development of a high-throughput LC/APCI-MS method for the determination of thirteen phytoestrogens including gut microbial metabolites in human urine and serum.

    Science.gov (United States)

    Wyns, Ciska; Bolca, Selin; De Keukeleire, Denis; Heyerick, Arne

    2010-04-15

    The investigation into the potential usefulness of phytoestrogens in the treatment of menopausal symptoms requires large-scale clinical trials that involve rapid, validated assays for the characterization and quantification of the phytoestrogenic precursors and their metabolites in biological matrices, as large interindividual differences in metabolism and bioavailability have been reported. Consequently, a new sensitive high-performance liquid chromatography-mass spectrometry method (HPLC-MS) for the quantitative determination of thirteen phytoestrogens including their most important gut microbial metabolites (genistein, daidzein, equol, dihydrodaidzein, O-desmethylangolensin, coumestrol, secoisolariciresinol, matairesinol, enterodiol, enterolactone, isoxanthohumol, xanthohumol and 8-prenylnaringenin) in human urine and serum within one single analytical run was developed. The method uses a simple sample preparation procedure consisting of enzymatic deconjugation followed by liquid-liquid extraction (LLE) or solid-phase extraction (SPE) for urine or serum, respectively. The phytoestrogens and their metabolites are detected with a single quadrupole mass spectrometer using atmospheric pressure chemical ionization (APCI), operating both in the positive and the negative mode. This bioanalytical method has been fully validated and proved to allow an accurate and precise quantification of the targeted phytoestrogens and their metabolites covering the lower parts-per-billion range for the measurement of relevant urine and serum levels following ingestion of phytoestrogen-rich dietary supplements.

  15. Development of a cloud point extraction and spectrophotometry-based microplate method for the determination of nitrite in human urine and blood.

    Science.gov (United States)

    Zhao, Jiao; Lu, Yunhui; Fan, Chongyang; Wang, Jun; Yang, Yaling

    2015-02-05

    A novel and simple method for the sensitive determination of trace amounts of nitrite in human urine and blood has been developed by combination of cloud point extraction (CPE) and microplate assay. The method is based on the Griess reaction and the reaction product is extracted into nonionic surfactant Triton-X114 using CPE technique. In this study, decolorization treatment of urine and blood was applied to overcome the interference of matrix and enhance the sensitivity of nitrite detection. Multi-sample can be simultaneously detected thanks to a 96-well microplate technique. The effects of different operating parameters such as type of decolorizing agent, concentration of surfactant (Triton X-114), addition of (NH4)2SO4, extraction temperature and time, interfering elements were studied and optimum conditions were obtained. Under the optimum conditions, a linear calibration graph was obtained in the range of 10-400 ng mL(-1) of nitrite with limit of detection (LOD) of 2.5 ng mL(-1). The relative standard deviation (RSD) for determination of 100 ng mL(-1) of nitrite was 2.80%. The proposed method was successfully applied for the determination of nitrite in the urine and blood samples with recoveries of 92.6-101.2%.

  16. Determination of palladium in human urine by high-performance liquid chromatography and ultraviolet detection after ultraviolet photolysis and selective solid-phase extraction.

    Science.gov (United States)

    Philippeit, G; Angerer, J

    2001-09-05

    The high-performance liquid chromatographic method with UV detection described below permits the selective determination of traces of palladium in human urine. After UV photolysis, during which the complete organic matrix was destroyed, the palladium was selectively enriched by solid-phase extraction (SPE). The reversed-phase C18 SPE column material was loaded with the ligand N,N-diethyl-N'-benzoylthiourea (DEBT) which shows an excellent complexing capacity for palladium in acidic solutions and at room temperature. The Pd(DEBT)2 complex was eluted with ethanol. After isocratic separation on the analytical column (MeOH/H2O 98:2 (v/v)), the complex was detected at 274 nm. The detection limit was 10 ng Pd/l. The relative standard deviations (RSD) of the within-series imprecision were in the range between 11% (75 ng Pd/l) and 7% (180 ng Pd/l). The between-day imprecision was 11% (75 ng Pd/l) and 5% (180 ng Pd/l). The recovery rates ranged between 94 and 96%. Using this method, urine samples of 44 persons from the general population were analysed. Only in one urine sample could palladium be detected. For comparison, 10 persons with occupational palladium exposure were examined. The urinary concentrations ranged from <10 to 2,538 ng/l.

  17. A sensitive enzyme immunoassay for human epidermal growth factor. Determination of hEGF in human serum and urine and pharmacokinetics in mouse.

    Science.gov (United States)

    Hayashi, T; Hashimoto, K; Sakamoto, S

    1989-07-01

    A sensitive enzyme immunoassay for human epidermal growth factor (hEGF) is described. The anti-hEGF antibody was prepared by immunizing rabbits with hEGF, which was synthesized by Escherichia coli using the genetic engineering technique. The present assay system was based on the sandwiching of an antigen between anti-hEGF F(ab')2 precoated on a 96-well polystyrene plate and beta-D-galactosidase-labeled anti-hEGF Fab'. The range of measurable hEGF by this assay was 0.1-100 pg/well. Recoveries of hEGF added to serum and urine ranged between 94 and 108%. The intra- and inter-assay coefficients of variation were less than 6 and 8%, respectively. The results obtained by this assay method correlated well with those obtained by the radioimmunoassay method. By using this assay, the time course of serum hEGF levels in mice after the various administrations were also examined.

  18. Urine - abnormal color

    Science.gov (United States)

    ... medlineplus.gov/ency/article/003139.htm Urine - abnormal color To use the sharing features on this page, please enable JavaScript. The usual color of urine is straw-yellow. Abnormally colored urine ...

  19. [Preparation of molecularly imprinted polymers using dummy template and the applications in selective solid-phase extraction of seven bisphenols from human urine, bovine serum and beer samples].

    Science.gov (United States)

    Yang, Jiajia; Li, Yun; Wang Jinchengt; Sun, Xiaoli; Chen, Jiping

    2015-05-01

    A molecularly imprinted polymer (MIP) for selective recognition of seven bisphenols (BPs) was prepared using dummy template phenolphthalein (PP) by bulk polymerization. The characterization results of scanning electron microscopy and nitrogen adsorption-desorption measurements showed that the prepared PP-MIP possessed narrow particle diameter distribution (40-60 µm), a specific surface area (S(BET)) of 359.77 m2/g and a total pore volume (Vt) of 0.730 cm3/g. The adsorption capacity for bisphenol A (BPA) of PP-MIP was evaluated by static adsorption experiment. And the Scatchard analysis revealed that the maximum specific adsorption capacity of PP-MIP was 4.661 µmol/g. Good class selectivity for BPA and its six structural analogues of bisphenol B (BPB) , bisphenol E (BPE), bisphenol AF (BPAF), bisphenol S (BPS), bisphenol AP (BPAP) and bisphenol Z (BPZ) was demonstrated by the chromatographic evaluation experiment. The prepared PP-MIP was successfully used as solid-phase extraction (SPE) sorbent for the separation and purification of the seven BPs in human urine, bovine serum and beer samples. Meanwhile, an accurate and sensitive MIP-SPE-HPLC method was established for the determination of the seven BPs in human urine, bovine serum and beer samples. The limits of detection (LODs) for the three samples were in the range of 1.2-2.0 µg/L. The results showed that good linearities were obtained in the range of 0.02-2 mg/L for the seven BPs and the correlation coefficients (r) were higher than 0.999 8. The recoveries of the BPs spiked in blank samples at two spiked levels (100 and 500 µg/L for each BP) were in the range of 90.1%-107.1% with the RSDs ≤ 8.1%. The proposed method is simple and reliable for the rapid detection of the seven BPs in human urine, bovine serum and beer samples.

  20. Simultaneous determination of blonanserin and its metabolite in human plasma and urine by liquid chromatography-tandem mass spectrometry: application to a pharmacokinetic study.

    Science.gov (United States)

    Wen, Yu-Guan; Ni, Xiao-Jia; Zhang, Ming; Liu, Xia; Shang, De-Wei

    2012-08-15

    Blonanserin is a novel atypical antipsychotic with highly selective receptor antagonist activity to dopamine D₂ and 5-HT(2A). N-desethyl blonanserin (blonanserin C) is its major active metabolite in human plasma. Herein we report a new highly sensitive, selective, and rapid liquid chromatography-tandem mass spectrometry method to determine blonanserin and blonanserin C simultaneously in human plasma and urine, with N-desethyl-chlor-blonanserin (blonanserin D) as internal standard (IS). Blonanserin and blonanserin C were extracted from aliquots of plasma and urine with ethyl acetate and dichloromethane (4:1) as the solvent and chromatographic separation was performed using an Agilent Eclipse Plus C₁₈ column. The mobile phase was composed of: acetonitrile and ammonium formate-formic acid buffer containing 5mM ammonium formate and 0.1% formic acid (87:13, v/v). To quantify blonanserin, blonanserin C, and blonanserin D, respectively, multiple reaction monitoring (MRM) transition of m/z 368.2→297.2, m/z 340.2→297.1, and m/z 356.2→313.3 was performed in positive mode. The analysis time was about 5.5 min. The calibration curve was linear in the concentration range of 0.01-2 ng/ml. The lower limit of quantification reached 0.01 ng/ml. The intra and inter-day precision and relative errors were less than 8.0% and 6.6% for three QC levels in plasma and urine. The current LC-MS/MS method was validated as simple, sensitive, and accurate and has been successfully applied to investigate the pharmacokinetics of blonanserin and blonanserin C in humans. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. The electroanalysis of mannitol, xylose and lactulose at copper electrodes: voltammetric studies and bioanalysis in human urine by means of HPLC with electrochemical detection.

    Science.gov (United States)

    Wring, S A; Terry, A; Causon, R; Jenner, W N

    1998-03-01

    The electrochemical behavior of mannitol, xylose and lactulose has been investigated at a copper working electrode. A sensitive, accurate and precise method employing HPLC with electrochemical detection in the d.c. amperometric mode, has been developed and validated for the determination of mannitol and lactulose in human urine. The ratio of these probe carbohydrates is altered in conditions that cause damage to the intestinal mucosal barrier. Systematic studies employing cyclic voltammetry indicate that the electrode reaction involves an electrocatalytic oxidation of each carbohydrate in a process yielding a single irreversible anodic wave that is dependent on the ionic strength of the sodium hydroxide supporting electrolyte solution. High performance liquid chromatography with electrochemical detection was performed using a thin-layer cell housing a custom manufactured copper working electrode. The optimized HPLC method can detect 72, 57 and 419 pg of mannitol, xylose and lactulose injected on column, respectively. The corresponding linear calibration ranges are 359 pg-2.24 microgram, 57.4 pg-896 ng and 419 pg-262 ng, respectively. Solid-phase extraction of human urine on polar sorbents, and direct injection after simple 1 + 99 dilution in 0.025 M NaOH were compared for bioanalysis. Direct injection was selected for further method developed as the technique proved robust and simple. The optimized method was validated for the determination of mannitol and lactulose in human urine over the concentration ranges predicted when assessing intestinal permeability (0.25-2.5 mg ml-1 mannitol and 0.05-1.0 mg ml-1 lactulose). Over these ranges intra- and inter-assay bias is < +/- 6.5%, and imprecision (coefficient of variation) is < 9% for each carbohydrate. The validated method provides a useful alternative to HPLC with pulsed-amperometric detection at gold electrodes.

  2. Analytical sample preparation strategies for the determination of antimalarial drugs in human whole blood, plasma and urine

    DEFF Research Database (Denmark)

    Casas, Monica Escolà; Hansen, Martin; Krogh, Kristine A;

    2014-01-01

    Antimalarial drugs commonly referred to as antimalarials, include a variety of compounds with different physicochemical properties. There is a lack of information on antimalarial distribution in the body over time after administration, e.g. the drug concentrations in whole blood, plasma, and urin...... summarized. Finally, the main problems that the researchers have dealt with are highlighted. This information will aid analytical chemists in the development of novel methods for determining existing antimalarials and upcoming new drugs.......Antimalarial drugs commonly referred to as antimalarials, include a variety of compounds with different physicochemical properties. There is a lack of information on antimalarial distribution in the body over time after administration, e.g. the drug concentrations in whole blood, plasma, and urine...

  3. Proteomic analysis of proteins selectively associated with hydroxyapatite, brushite, and uric acid crystals precipitated from human urine.

    Science.gov (United States)

    Thurgood, Lauren A; Ryall, Rosemary L

    2010-10-01

    The aim of this study was to compare the intracrystalline protein profiles of hydroxyapatite (HA), brushite (BR), and uric acid (UA) crystals precipitated from the same urine samples. HA, BR, and UA crystals were precipitated on two different occasions from the same pooled healthy urine. Crystals were washed to remove surface-bound proteins, and their composition was confirmed using Fourier transform infrared spectroscopy (FTIR) and field emission scanning electron microscopy (FESEM) coupled with energy dispersive X-ray analysis (EDAX). SDS-PAGE was used for visual comparison of the protein content of the demineralised crystal extracts, which were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). HA comprised nanosized particles interspersed with organic material, which was absent from the BR and UA crystals. The number and type of individual proteins differed between the 3 minerals: 45 proteins were detected in the HA crystal extracts and 77 in the BR crystals, including a number of keratins, which were regarded as methodological contaminants. After excluding the keratins, 21 proteins were common to both HA and BR crystals. Seven nonkeratin proteins were identified in the UA extracts. Several proteins consistently detected in the HA and BR crystal extracts have been previously implicated in kidney stone disease, including osteopontin, prothrombin, protein S100A9 (calgranulin B), inter-α-inhibitor, α1-microglobulin bikunin (AMBP), heparan sulfate proteoglycan, and Tamm-Horsfall glycoprotein, all of which are strong calcium binders. We concluded that the association of proteins with HA, BR, and UA crystals formed in healthy urine is selective and that only a few of the numerous proteins present in healthy urine are likely to play any significant role in preventing stone pathogenesis.

  4. Fluoride in drinking water, brick tea infusion and human urine in two counties in Inner Mongolia, China.

    Science.gov (United States)

    Li, Hai-rong; Liu, Qing-bin; Wang, Wu-yi; Yang, Lin-sheng; Li, Yong-hua; Feng, Fu-jian; Zhao, Xiao-yu; Hou, Kun; Wang, Ge

    2009-08-15

    The objective of this study was to detect the fluoride level in the drinking water and the urine of habitants aged 16-55 years living in Inner Mongolia China. Furthermore, fluoride concentration of the brick tea infusion samples which were drunk by Mongolia herdsmen in everyday life living in SumuErga village of Ejin Horo Banner, Inner Mongolia China was also determined. A total of 117 participants (61 female and 56 male) were recruited from two counties for a cross-sectional study on health effects of chronic fluoride exposure from drinking water and drinking brick tea infusion. The fluoride concentration in drinking water, urine and brick tea infusion samples were determined using fluoride ion selective electrode method obtained from the Ministry of Health of the People's Republic of China. The average fluoride concentration in drinking water samples was 0.32+/-0.01 mg/L at AretengXire town of Ejin Horo Banner, 0.70+/-0.19 mg/L at SumuErga village of Ejin Horo Banner, and 2.68+/-1.15 mg/L at ZhalaiNuoer district of Manzhouli city. The average fluoride concentration in brick tea infusion samples which collected from Mongolia herdsmen at SumuErga village of Ejin Horo Banner was 1.81+/-1.09 mg/L. The average urinary fluoride concentration at AretengXire town of Ejin Horo Banner was 0.59+/-0.48 mg/L, at SumuErga village of Ejin Horo Banner was 1.45+/-0.93 mg/L and at ZhalaiNuoer district of Manzhouli city was 3.06+/-1.53 mg/L. The higher fluoride levels in the urine of participants may be associated to higher fluoride in drinking water at ZhalaiNuoer of Manzhouli city. However, drinking brick tea infusions with higher fluoride may be the cause of the higher fluoride contents in the Mongolia herdsmen's urine.

  5. Carbon isotopes profiles of human whole blood, plasma, red blood cells, urine and feces for biological/biomedical 14C-accelerator mass spectrometry applications.

    Science.gov (United States)

    Kim, Seung-Hyun; Chuang, Jennifer C; Kelly, Peter B; Clifford, Andrew J

    2011-05-01

    Radiocarbon ((14)C) is an ideal tracer for in vivo human ADME (absorption, distribution, metabolism, elimination) and PBPK (physiological-based pharmacokinetic) studies. Living plants peferentially incorporate atmospheric (14)CO(2) versus (13)CO(2) versus (12)CO(2), which result in unique signature. Furthermore, plants and the food chains they support also have unique carbon isotope signatures. Humans, at the top of the food chain, consequently acquire isotopic concentrations in the tissues and body fluids depending on their dietary habits. In preparation of ADME and PBPK studies, 12 healthy subjects were recruited. The human baseline (specific to each individual and their diet) total carbon (TC) and carbon isotope (13)C (δ(13)C) and (14)C (F(m)) were quantified in whole blood (WB), plasma, washed red blood cell (RBC), urine, and feces. TC (mg of C/100 μL) in WB, plasma, RBC, urine, and feces were 11.0, 4.37, 7.57, 0.53, and 1.90, respectively. TC in WB, RBC, and feces was higher in men over women, P < 0.05. Mean δ(13)C were ranked low to high as follows: feces < WB = plasma = RBC = urine, P < 0.0001. δ(13)C was not affected by gender. Our analytic method shifted δ(13)C by only ±1.0 ‰ ensuring our F(m) measurements were accurate and precise. Mean F(m) were ranked low to high as follows: plasma = urine < WB = RBC = feces, P < 0.05. F(m) in feces was higher for men over women, P < 0.05. Only in WB, (14)C levels (F(m)) and TC were correlated with one another (r = 0.746, P < 0.01). Considering the lag time to incorporate atmospheric (14)C into plant foods (vegetarian) and or then into animal foods (nonvegetarian), the measured F(m) of WB in our population (recruited April 2009) was 1.0468 ± 0.0022 (mean ± SD), and the F(m) of WB matched the (extrapolated) atmospheric F(m) of 1.0477 in 2008. This study is important in presenting a procedure to determine a baseline for a study group for human ADME and PBPK studies using (14)C as a tracer.

  6. Hydrophilic interaction chromatography combined with dispersive liquid-liquid microextraction as a preconcentration tool for the simultaneous determination of the panel of underivatized neurotransmitters in human urine samples.

    Science.gov (United States)

    Konieczna, Lucyna; Roszkowska, Anna; Niedźwiecki, Maciej; Bączek, Tomasz

    2016-01-29

    A simple and sensitive method using dispersive liquid-liquid microextraction (DLLME) followed by liquid chromatography coupled to mass spectrometry (LC-MS) with a hydrophilic interaction chromatography (HILIC) column was developed for the simultaneous determination of 13 compounds of different polarities, comprising monoamine neurotransmitters (dopamine, norepinephrine, epinephrine and serotonin) along with their respective precursors and metabolites, in human urine samples. The microextraction procedure was based on the fast injection of a mixture of ethanol (disperser solvent) and dichloromethane (extraction solvent) into a human urine sample, forming a cloudy solution in the Eppendorf tube. After centrifugation, the sedimented phase was collected and subsequently analyzed by LC-HILIC-MS in about 12min without a derivatization step. The separation was performed on an XBridge Amide™ BEH column 3.0×100mm, 3.5mm and the mobile phase consisted of phase A: 10mM ammonium formate buffer in water pH 3.0 and phase B: 10 mM ammonium formate buffer in acetonitrile, under gradient program elution. Tyrosine, tryptophan, 5-hydroxytryptophan, dopamine, epinephrine, norepinephrine, serotonin, 3-methoxytyramine, 5-hydroxyindole-3-acetic acid, 3,4-dihydroxy-l-phenylalanine and norvaline (internal standard) were detected in the positive ionization mode. While vanillylmandelic acid, homovanillic acid, 3,4-dihydroxyphenylacetic acid and 3,4-dihydroxybenzylamine (internal standard) were detected in the negative ionization mode. Parameters influencing DLLME and LC-HILIC-MS were investigated. Under the optimum conditions, the proposed method exhibited a low detection limit (5-10ngmL(-1)), and good linearity with R between 0.9991 and 0.9998. The recoveries in human urine samples were 99.0%±3.6%. for the 13 studied biogenic amines with intra- and inter-day RSDs of 0.24-9.55% and 0.31-10.0%, respectively. The developed DLLME-LC-MS method could be successfully applied for the

  7. Indacaterol determination in human urine: validation of a liquid-liquid extraction and liquid chromatography-tandem mass spectrometry analytical method.

    Science.gov (United States)

    Ammari, Wesam G; Al-Qadhi, Zainab; Khalil, Mohammad; Tayyem, Rabab; Qammaz, Samir; Oriquat, Ghaleb; Basheti, Iman A; Chrystyn, Henry

    2015-06-01

    Indacaterol is a novel once-a-day inhaled ultra-long-acting β2-agonist. Quantitative bioanalysis supports pharmacokinetic and clinical research. The aim of the current work was to validate an in-house developed high performance liquid chromatography (HPLC)-tandem mass spectrometry (MS/MS) analytical method for indacaterol determination in human urine samples. A liquid-liquid extraction method has been developed to extract indacaterol from human urine samples using ethyl acetate. Indacaterol dry extract was reconstituted with 200 μL of the mobile phase (acidified water:methanol (30:70, v/v)) of which 5 μL was needed for the HPLC-MS/MS analysis. Indacaterol was eluted on a reversed C18 stationary phase with an isocratic mobile phase at a flow of 1 mL/min. Formoterol was the internal standard (IS). The MS/MS detection was employed with a turbo-ion spray ionization in the positive ion mode. A consensus of the international Guidelines for Bioanalytical Method Validation was followed. Indacaterol was detected at a mass to charge ratio (m/z) of 393.3 and its MS/MS daughter at 173.2. The retention times of indacaterol and IS were 1.60 and 1.20 min, respectively. Validated calibration curves were linear over a range of 0.075-100 ng/mL with correlation coefficients (r)≥0.990. The curves' regression weighting factor was 1/x. Method specificity was established in six different human urine batches. No matrix interference was observed. The intra- and inter-batch precision and accuracy within±20% (at lower limit) and±15% (other quality control (QC) levels) were confirmed. The indacaterol mean recovery (precision) percentages at Low, Mid, and High QC levels were 93.5 (3.84), 89.8 (2.15), and 92.2 (2.17), respectively. Short-term, long-term, freeze-thaw, and auto-sampler stability results were accepted. A specific, accurate and precise HPLC-MS/MS method has been validated for indacaterol quantification in human urine. This simple method is reproducible and robust

  8. Identification and quantification of flavonoids in human urine samples by column switching liquid chromatography coupled to atmospheric pressure chemical ionization mass spectrometry

    DEFF Research Database (Denmark)

    Nielsen, Salka E.; Freese, R.; Cornett, C.

    2000-01-01

    A rapid and sensitive high-performance liquid chromatographic mass spectrometric (HPLC-MS) method is described for the determination and quantification of 12 dietary flavonoid glycosides and aglycons in human urine samples. Chromatographic separation of the analytes of interest was achieved...... by column-switching, using the first column (a Zorbax 300SB C-3 column) for sample cleanup and eluting the heart-cut flavonoid fraction onto the second column (a Zorbax SE C-18 column) for separation and detection by ultraviolet and atmospheric pressure chemical ionization MS using single ion monitoring...... with high and low flavonoid content was analyzed, and the results are reported....

  9. Variation in Gas and Volatile Compound Emissions from Human Urine as It Ages, Measured by an Electronic Nose

    Directory of Open Access Journals (Sweden)

    Siavash Esfahani

    2016-01-01

    Full Text Available The medical profession is becoming ever more interested in the use of gas-phase biomarkers for disease identification and monitoring. This is due in part to its rapid analysis time and low test cost, which makes it attractive for many different clinical arenas. One technology that is showing promise for analyzing these gas-phase biomarkers is the electronic nose—an instrument designed to replicate the biological olfactory system. Of the possible biological media available to “sniff”, urine is becoming ever more important as it is easy to collect and to store for batch testing. However, this raises the question of sample storage shelf-life, even at −80 °C. Here we investigated the effect of storage time (years on stability and reproducibility of total gas/vapour emissions from urine samples. Urine samples from 87 patients with Type 2 Diabetes Mellitus were collected over a four-year period and stored at −80 °C. These samples were then analyzed using FAIMS (field-asymmetric ion mobility spectrometry—a type of electronic nose. It was discovered that gas emissions (concentration and diversity reduced over time. However, there was less variation in the initial nine months of storage with greater uniformity and stability of concentrations together with tighter clustering of the total number of chemicals released. This suggests that nine months could be considered a general guide to a sample shelf-life.

  10. Micellar Enhanced Spectrofluorimetric Method for the Determination of Ponatinib in Human Plasma and Urine via Cremophor RH 40 as Sensing Agent

    Science.gov (United States)

    Darwish, Hany W.; Bakheit, Ahmed H.; Abdelhameed, Ali Saber; AlKhairallah, Amer S.

    2015-01-01

    An impressively simple and precise spectrofluorimetric procedure was established and validated for ponatinib (PTB) quantitation in biological fluids such as human plasma and human urine. This method depends on examining the fluorescence characteristics of PTB in a micellar system of Cremophor RH 40 (Cr RH 40). Cr RH 40 enhanced the intrinsic fluorescence of PTB distinctly in aqueous water. The fluorescence spectra of PTB was recorded at 457 nm following its excitation at 305 nm. Maximum fluorescence intensity was attained by addition of 0.7 mL of Cr RH 40 and one mL of phosphate buffer to PTB aliquots and then dilution with distilled water. There is a linear relationship between the fluorescence intensity of PTB and its concentration over the range 5–120 ngmL−1, with limit of detection and limit of quantification equal to 0.905 ngmL−1 and 2.742 ngmL−1, respectively. The accuracy and the precisions of the proposed method were checked and gave adequate results. The adopted method was applied with a great success for PTB quantitation in different biological matrices (spiked human plasma and urine) giving high recovery values. PMID:26880920

  11. Utilización de excretas de las aves en alimentación de rumiantes

    OpenAIRE

    2013-01-01

    Utilization of poultry excreta in rumiant feeding. A study wasconducted to determine chemical composition of poultry escreta in Costa Rica, artd a/so is discurs the use and limitation of these products in ruminant f eeding. Fesu/fs sho wed that the limitant f actor of these by.products is the catcium content; which was 3,1a and 6.13% for broiler titter and tayer litter respectively. Because of this, is not recommend the use of layer manure in ruminant feedinE' The nutrients content studied ar...

  12. Willingness to pay for excreta pellet fertilizer: Empirical evidence from Ghana

    Directory of Open Access Journals (Sweden)

    JOHN K. M. Kuwornu

    2017-09-01

    Full Text Available This study examined farmers’ willingness to pay for excreta pellet fertilizer in Ghana. Primary data was obtained from 461 farmers in 10 districts in the Western and Greater Accra regions of Ghana through randomized questionnaire administration. The contingent valuation method was used in eliciting the farmers’ willingness to pay decisions (WTP and maximum amount they are willing to pay. The Tobit regression model results revealed that being a household head, unit cost of current fertilizer used, and farm size positively influenced the willingness to pay amount whereas previous use of organic fertilizer influenced the willingness to pay amount negatively.

  13. Urine Protein and Urine Protein to Creatinine Ratio

    Science.gov (United States)

    ... products and services. Advertising & Sponsorship: Policy | Opportunities Urine Protein and Urine Protein to Creatinine Ratio Share this page: Was this page helpful? Also known as: 24-Hour Urine Protein; Urine Total Protein; Urine Protein to Creatinine Ratio; ...

  14. Development and validation of an ultra-high performance liquid chromatography-tandem mass spectrometry method to measure creatinine in human urine.

    Science.gov (United States)

    Fraselle, S; De Cremer, K; Coucke, W; Glorieux, G; Vanmassenhove, J; Schepers, E; Neirynck, N; Van Overmeire, I; Van Loco, J; Van Biesen, W; Vanholder, R

    2015-04-15

    Despite decades of creatinine measurement in biological fluids using a large variety of analytical methods, an accurate determination of this compound remains challenging. Especially with the novel trend to assess biomarkers on large sample sets preserved in biobanks, a simple and fast method that could cope with both a high sample throughput and a low volume of sample is still of interest. In answer to these challenges, a fast and accurate ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed to measure creatinine in small volumes of human urine. In this method, urine samples are simply diluted with a basic mobile phase and injected directly under positive electrospray ionization (ESI) conditions, without further purification steps. The combination of an important diluting factor (10(4) times) due to the use of a very sensitive triple quadrupole mass spectrometer (XEVO TQ) and the addition of creatinine-d3 as internal standard completely eliminates matrix effects coming from the urine. The method was validated in-house in 2012 according to the EMA guideline on bioanalytical method validation using Certified Reference samples from the German External Quality Assessment Scheme (G-Equas) proficiency test. All obtained results for accuracy and recovery are within the authorized tolerance ranges defined by G-Equas. The method is linear between 0 and 5 g/L, with LOD and LOQ of 5 × 10(-3) g/L and 10(-2) g/L, respectively. The repeatability (CV(r) = 1.03-2.07%) and intra-laboratory reproducibility (CV(RW) = 1.97-2.40%) satisfy the EMA 2012 guideline. The validated method was firstly applied to perform the German G-Equas proficiency test rounds 51 and 53, in 2013 and 2014, respectively. The obtained results were again all within the accepted tolerance ranges and very close to the reference values defined by the organizers of the proficiency test scheme, demonstrating an excellent accuracy of the developed method. The

  15. Quantitative determination of BAF312, a S1P-R modulator, in human urine by LC-MS/MS: prevention and recovery of lost analyte due to container surface adsorption.

    Science.gov (United States)

    Li, Wenkui; Luo, Suyi; Smith, Harold T; Tse, Francis L S

    2010-02-15

    Analyte loss due to non-specific binding, especially container surface adsorption, is not uncommon in the quantitative analysis of urine samples. In developing a sensitive LC-MS/MS method for the determination of a drug candidate, BAF312, in human urine, a simple procedure was outlined for identification, confirmation and prevention of analyte non-specific binding to a container surface and to recover the 'non-specific loss' of an analyte, if no transfer has occurred to the original urine samples. Non-specific binding or container surface adsorption can be quickly identified by using freshly spiked urine calibration standards and pre-pooled QC samples during a LC-MS/MS feasibility run. The resulting low recovery of an analyte in urine samples can be prevented through the use of additives, such as the non-ionic surfactant Tween-80, CHAPS and others, to the container prior to urine sample collection. If the urine samples have not been transferred from the bulk container, the 'non-specific binding' of an analyte to the container surface can be reversed by the addition of a specified amount of CHAPS, Tween-80 or bovine serum albumin, followed by appropriate mixing. Among the above agents, Tween-80 is the most cost-effective. beta-cyclodextrin may be suitable in stabilizing the analyte of interest in urine via pre-treating the matrix with the agent. However, post-addition of beta-cyclodextrin to untreated urine samples does not recover the 'lost' analyte due to non-specific binding or container surface adsorption. In the case of BAF312, a dynamic range of 0.0200-20.0 ng/ml in human urine was validated with an overall accuracy and precision for QC sample results ranging from -3.2 to 5.1% (bias) and 3.9 to 10.2% (CV), respectively. Pre- and post-addition of 0.5% (v/v) Tween-80 to the container provided excellent overall analyte recovery and minimal MS signal suppression when a liquid-liquid extraction in combination with an isocratic LC separation was employed. The

  16. Dispersive liquid-liquid microextraction based on solidification of floating organic drop and high-performance liquid chromatography to the analysis of cocaine's major adulterants in human urine.

    Science.gov (United States)

    Sena, Laís Cristina Santana; Matos, Humberto Reis; Dórea, Haroldo Silveira; Pimentel, Maria Fernanda; de Santana, Danielle Cristine Almeida Silva; de Santana, Fernando José Malagueño

    2017-02-01

    A simple method has been proposed for the determination of cocaine's major adulterants (caffeine, levamisole, lidocaine, phenacetin, diltiazem, and hydroxyzine) in human urine by dispersive liquid-liquid microextraction based on solidification of floating organic drop (DLLME-SFO) in combination with high-performance liquid chromatography - photodiode array detector (HPLC-PDA). The reversed-phase chromatographic separation was obtained with a column C18 extended (250×4.6mm; 5μm; 80Å) in gradient elution mode using acetonitrile-trifluoroacetic acid 0.026% (v,v) (pH=2.5) at 1mLmin(-1) as mobile phase, at 25°C, and detection at 235nm. The analysis time was 25min. This condition had the best resolution factors (>1.15), retention factors (>0.68), number of plates (>2094.9), and separation factors (>1.05) for all targets, indicating a good separation. The kind of extraction and dispersive solvent were investigated for unifactorial design. The buffer pH, the volume of extraction and disperser solvent, and the amount of salt were optimized for full factorial design. Under optimum conditions, human urine samples were alkalized with 0.5M sodium phosphate buffer (pH 10) and added to sodium chloride (20%m/v). Acetonitrile (150μL) and 1-dodecanol (30μL) were used as dispersive and extraction solvent, respectively. The method presented linear range of 312.5-3125ngmL(-1) to caffeine and levamisole and 187.5-1875ngmL(-1) to lidocaine, phenacetin, diltiazem, and hydroxyzine. The limit of quantification was 187.5ngmL(-1) to lidocaine, phenacetin, diltiazem, and hydroxyzine and 312.5ngmL(-1) for caffeine and levamisole. The recovery mean values were between 6.0 and 42.6%. The method showed good precision and accuracy, with within- and between-run relative standard deviation and relative error less than 15%. The samples were stable after freeze-thaw cycle and short-term room temperature stability tests. Besides, this method was satisfactorily applied in urine of cocaine users. It

  17. Molecularly imprinted polymer microspheres prepared by Pickering emulsion polymerization for selective solid-phase extraction of eight bisphenols from human urine samples

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Jiajia [Key Laboratory of Separation Sciences for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Li, Yun; Wang, Jincheng [Key Laboratory of Separation Sciences for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); Sun, Xiaoli; Cao, Rong [Key Laboratory of Separation Sciences for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Sun, Hao [Key Laboratory of Separation Sciences for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); Department of Chemistry, Liaoning University, Shenyang 110000 (China); Huang, Chaonan [Key Laboratory of Separation Sciences for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Chen, Jiping, E-mail: chenjp@dicp.ac.cn [Key Laboratory of Separation Sciences for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian 116023 (China)

    2015-05-04

    Highlights: • BPA imprinted polymer microspheres were prepared by Pickering emulsion polymerization. • Regular spherical shape and narrow diameter distribution. • Good specific adsorption capacity for BPA. • Good class-selectivity and clean-up efficiency for bisphenols in human urine under SPE mode. • Good recoveries and sensitivity for bisphenols using the MIPMS-SPE coupled with HPLC-DAD method. - Abstract: The bisphenol A (BPA) imprinted polymer microspheres were prepared by simple Pickering emulsion polymerization. Compared to traditional bulk polymerization, both high yields of polymer and good control of particle sizes were achieved. The characterization results of scanning electron microscopy and nitrogen adsorption–desorption measurements showed that the obtained molecularly imprinted polymer microsphere (MIPMS) particles possessed regular spherical shape, narrow diameter distribution (30–60 μm), a specific surface area (S{sub BET}) of 281.26 m{sup 2} g{sup −1} and a total pore volume (V{sub t}) of 0.459 cm{sup 3} g{sup −1}. Good specific adsorption capacity for BPA was obtained in the sorption experiment and good class selectivity for BPA and its seven structural analogs (bisphenol F, bisphenol B, bisphenol E, bisphenol AF, bisphenol S, bisphenol AP and bisphenol Z) was demonstrated by the chromatographic evaluation experiment. The MIPMS as solid-phase extraction (SPE) packing material was then evaluated for extraction and clean-up of these bisphenols (BPs) from human urine samples. An accurate and sensitive analytical method based on the MIPMS-SPE coupled with HPLC-DAD has been successfully established for simultaneous determination of eight BPs from human urine samples with detection limits of 1.2–2.2 ng mL{sup −1}. The recoveries of BPs for urine samples at two spiking levels (100 and 500 ng mL{sup −1} for each BP) were in the range of 81.3–106.7% with RSD values below 8.3%.

  18. Determination of 2,3-dihydroxypropionamide, an oxidative metabolite of acrylamide, in human urine by gas chromatography coupled with mass spectrometry.

    Science.gov (United States)

    Latzin, Julia M; Schindler, Birgit K; Weiss, Tobias; Angerer, Jürgen; Koch, Holger M

    2012-03-01

    The general population is exposed to acrylamide (AA) mainly through food and tobacco smoke. AA is classified as probably carcinogenic to humans. Glycidamide (GA), as the primary oxidative metabolite, was identified to be the ultimate genotoxic agent. This warrants full investigation of the oxidative pathway in AA metabolism and the share of the oxidative compared to the reductive pathway. 2,3-Dihydroxy-propionamide (OH-PA) as the direct hydrolysis product of GA has been shown to be a major urinary oxidative metabolite in human AA metabolism. We developed an analytical method to reliably quantify OH-PA in urine by GC-MS after a multistep procedure including "stripping" on a solid phase material, lyophilization, silylation and re-extraction. With a detection limit of 1 μg/L, our method is sensitive enough to quantify OH-PA in all urine samples of the general population. Within and between series precisions were between 1.9% and 8.2% and mean recoveries between 97% and 101%. We applied this method to 30 urine samples from the general population. In all the samples, OH-PA was present in concentrations between 6.8 and 109.4 μg/L (median, 49.7 μg/L) with no difference between smokers and non-smokers. OH-PA concentrations were approximately ten times higher than expected from the metabolism of AA via GA. Currently, we cannot confirm OH-PA to be a specific biomarker of the oxidative pathway of AA metabolism. Other sources than AA respectively GA might need to be considered for the formation of OH-PA.

  19. Transcriptional analysis of the Escherichia coli ColV-Ia plasmid pS88 during growth in human serum and urine

    Directory of Open Access Journals (Sweden)

    Lemaître Chloé

    2012-06-01

    Full Text Available Abstract Background The sequenced O45:K1:H7 Escherichia coli meningitis strain S88 harbors a large virulence plasmid. To identify possible genetic determinants of pS88 virulence, we examined the transcriptomes of 88 plasmidic ORFs corresponding to known and putative virulence genes, and 35 ORFs of unknown function. Results Quantification of plasmidic transcripts was obtained by quantitative real-time reverse transcription of extracted RNA, normalized on three housekeeping genes. The transcriptome of E. coli strain S88 grown in human serum and urine ex vivo were compared to that obtained during growth in Luria Bertani broth, with and without iron depletion. We also analyzed the transcriptome of a pS88-like plasmid recovered from a neonate with urinary tract infection. The transcriptome obtained after ex vivo growth in serum and urine was very similar to those obtained in iron-depleted LB broth. Genes encoding iron acquisition systems were strongly upregulated. ShiF and ORF 123, two ORFs encoding protein with hypothetical function and physically linked to aerobactin and salmochelin loci, respectively, were also highly expressed in iron-depleted conditions and may correspond to ancillary iron acquisition genes. Four ORFs were induced ex vivo, independently of the iron concentration. Other putative virulence genes such as iss, etsC, ompTp and hlyF were not upregulated in any of the conditions studied. Transcriptome analysis of the pS88-like plasmid recovered in vivo showed a similar pattern of induction but at much higher levels. Conclusion We identify new pS88 genes potentially involved in the growth of E. coli meningitis strain S88 in human serum and urine.

  20. [Simultaneous determination of five active components of compound α-ketoacid tablet in human urine by ion-pair reversed-phase high performance liquid chromatography].

    Science.gov (United States)

    Huang, Xiaoya; Zhong, Yuan; Huang, Zhongping; Jin, Chen; Wang, Lili; Pan, Zaifa

    2015-02-01

    A simple and sensitive method for the simultaneous determination of five active components, D, L-α-hydroxymethionine calcium (HMACa), α-ketovaline calcium (KVCa), D, L-α-ketoisoleucine calcium (KILCa), α-ketoleucine calcium (KLCa) and α-ketophenylalanine calcium (KPACa) of compound α-ketoacid tablet in human urine by ion-pair reversed-phase high performance liquid chromatography (RP-HPLC) was developed and validated. The separation conditions, such as the concentration of ion-pair reagent, the pH value of the mobile phase and the concentration of the buffer were optimized. All the five analytes were separated well on a C18 column (250 mm x 4.6 mm, 5 µm) with diode array detection at 210 nm and the column temperature of 35 °C. The mobile phases were acetonitrile and 20 mmol/L phosphate buffer (containing 15 mmol/L tetrabutylammonium hydroxide; pH 7) at the flow rate of 1. 0 mL/min with gradient elution. The calibration curves for the five components were linear in the range from 20 to 200 mg/L (r≥ 0. 9990). The limits of detection (LODs, S/N= 3) were 3.0, 5. 0, 3. 6, 5.7 and 2. 5 mg/L, and the limits of quantification (LOQs, S/N= 10) were 9. 6, 16.7, 12.0, 19.0 and 8.3 mg/L for HMACa, KVCa, KILCa, KLCa and KPACa, respectively. The intra-day and inter-day precisions were less than 7%, and the average recoveries were between 86.79% and 112. 00% in the human urine with RSDs lower than 9% (n= 5). The method proved precise, specific and reproducible, and can be used for the determination of the five components in urine.

  1. Monitoring of urine by extraction chromatography with tri-n-octylphosphine oxide supported on a polypropylene column

    Energy Technology Data Exchange (ETDEWEB)

    Bonino, N.; Diodati, J.; Cena, M.R. [Gerencia de Seguridad Radiologica y Nuclear, Comision Nacional de Energia Atomica, Buenos Aires (Argentina)

    1992-07-01

    Monitoring of personnel working with 20% enriched uranium implies development of techniques for excreta analysis, mainly for urine, with very low detection limits. The method described allows the determination of 20% enriched uranium after extraction in tri-n-octylphosphine oxide (TOPO), 0.5 in toluene, supported on polypropylene capillary columns. Alpha activity is later measured in a low background liquid scintillation equipment and the fluorescence in a fluorimeter, with detection limits, for 800 mL of urine, of 15.0 {+-} 4.0 mBq L{sup -1} and 5x10{sup -2} {+-} 10{sup -2} {mu}g L{sup -1}. (author)

  2. Photoluminescence of urine salts

    Science.gov (United States)

    Bordun, O.; Drobchak, O.

    2008-02-01

    Photoexcitation and luminescence spectra of dried urine sample under laser excitation were studied. Luminescence spectra of urine are determined by luminescence of urea which is the main component of urine. The presence of pathological salts in urine leads to the long-wave shifting of maxima of luminescence and to the decreasing of luminescence intensity.

  3. Metabolism and elimination of methyl, iso- and n-butyl paraben in human urine after single oral dosage.

    Science.gov (United States)

    Moos, Rebecca K; Angerer, Jürgen; Dierkes, Georg; Brüning, Thomas; Koch, Holger M

    2016-11-01

    Parabens are used as preservatives in personal care and consumer products, food and pharmaceuticals. Their use is controversial because of possible endocrine disrupting properties. In this study, we investigated metabolism and urinary excretion of methyl paraben (MeP), iso-butyl paraben (iso-BuP) and n-butyl paraben (n-BuP) after oral dosage of deuterium-labeled analogs (10 mg). Each volunteer received one dosage per investigated paraben separately and at least 2 weeks apart. Consecutive urine samples were collected over 48 h. In addition to the parent parabens (free and conjugated) which are already used as biomarkers of internal exposure and the known but non-specific metabolites, p-hydroxybenzoic acid (PHBA) and p-hydroxyhippuric acid (PHHA), we identified new, oxidized metabolites with hydroxy groups on the alkyl side chain (3OH-n-BuP and 2OH-iso-BuP) and species with oxidative modifications on the aromatic ring. MeP represented 17.4 % of the dose excreted in urine, while iso-BuP represented only 6.8 % and n-BuP 5.6 %. Additionally, for iso-BuP, about 16 % was excreted as 2OH-iso-BuP and for n-BuP about 6 % as 3OH-n-BuP. Less than 1 % was excreted as ring-hydroxylated metabolites. In all cases, PHHA was identified as the major but non-specific metabolite (57.2-63.8 %). PHBA represented 3.0-7.2 %. For all parabens, the majority of the oral dose captured by the above metabolites was excreted in the first 24 h (80.5-85.3 %). Complementary to the parent parabens excreted in urine, alkyl-chain-oxidized metabolites of the butyl parabens are introduced as valuable and contamination-free biomarkers of exposure.

  4. Simultaneous determination of opiates, methadone, buprenorphine and metabolites in human urine by superficially porous liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Lin, Huei-Ru; Chen, Chin-Lun; Huang, Chieh-Liang; Chen, Shao-Tsu; Lua, Ahai-Chuang

    2013-04-15

    For monitoring compliance of methadone or buprenorphine maintenance patient, a method for the simultaneous determination of methadone, 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP), buprenorphine, norbuprenorphine, opiates (morphine, codeine, 6-monoacetylmorphine) in urine by superficially porous liquid chromatography tandem mass spectrometry was developed and validated. After enzyme digestion and liquid-liquid extraction, reverse-phase separation was achieved in 5.2 min and quantification was performed by multiple reaction monitoring. Chromatographic separation was performed at 40 °C on a reversed phase Poroshell column with gradient elution. The mobile phase consisted of water and methanol, each containing 0.1% formic acid, at a flow rate of 0.32 mL/min. Intra-day and inter-day precision were less than 12.1% and accuracy was between -9.8% and 13.7%. Extraction efficiencies were more than 68%. Although ion suppression was detected, deuterated internal standards compensated for these effects. Carryover was minimal, less than 0.20%. All analytes were stable at room temperature for 16 h, 4 °C for 72 h, and after three freeze-thaw cycles. The assay also fulfilled compound identification criteria in accordance with the European Commission Decision 2002/657/EC. We analyzed 62 urine samples from patients received maintenance therapy and found that 54.8% of the patient samples tested were detected for morphine, codeine, or 6-monoacetylmorphine. This method provides a reliable and simultaneous quantification of opiates, maintenance drugs, and their metabolites in urine samples. It facilitates the routine monitoring in individuals prescribed the drug to ensure compliance and help therapeutic process.

  5. Reassessment of 239Pu on planchets from human urine samples at ultra-trace levels using Aridus-ICPSFMS and AMS.

    Science.gov (United States)

    Hernández-Mendoza, Héctor; Chamizo, Elena; Delgado, Antonio; García-León, Manuel; Yllera, Abel

    2012-12-01

    New analytical methods developed at the facilities here, based on two ultra-sensitive mass spectrometry (MS) techniques, inductively coupled plasma sector field mass spectrometer with a desolvator system (Aridus-ICP-SFMS) and accelerator MS (AMS), have been applied in this work for the reassessment of (239)Pu in alpha spectrometry (AS) planchets corresponding to spiked human urine samples. The obtained (239)Pu minimum detectable activities (MDAs) values by Aridus-ICP-SFMS and AMS were 3 fg (∼6.92 µBq) and 0.4 fg (∼0.92 µBq), respectively, per sample, which are much better than those attainable by AS [50 fg (∼115.3 µBq) of (239)Pu per sample, approximately]. Therefore, it is demonstrated that the MS techniques employed in this work are very powerful tools for internal dosimetry studies in human urine samples, giving excellent results when the reassessment of AS planchets is needed (samples with a Pu concentration below or at the MDA levels measurable by AS). This work is the continuation of an article published in J. Anal. At. Spectrom. 25 (1410-1415) 2010.

  6. Visual and colorimetric detection of p-aminophenol in environmental water and human urine samples based on anisotropic growth of Ag nanoshells on Au nanorods.

    Science.gov (United States)

    Lin, Tianran; Li, Zhihong; Song, Zhiping; Chen, Huan; Guo, Liangqia; Fu, Fengfu; Wu, Zujian

    2016-01-01

    A simple, sensitive, selective and high-resolution colorimetric method has been developed for the detection of p-aminophenol in environmental water and human urine samples. In the presence of p-aminophenol, silver ions are reduced to silver atoms and subsequently Ag nanoshells anisotropically grow on the surface of Au nanorods to generate orange slice-like Au@Ag core-shell nanocrystals, thereby resulting in the blue-shift of longitudinal surface plasmon resonance band of Au nanorods accompanying a sharp-contrast multicolor change. Using Au@Ag core-shell nanocrystals as the transducer, sub-micromolar p-aminophenol can be detected by the colorimetric method and 10 μmol L(-1) p-aminophenol can be visual readout by the naked eyes. Furthermore, a simple, cheap, portable test kit is constructed for the visual assay of urinary p-aminophenol without complicated sample pretreatment and sophisticated instruments. The proposed colorimetric method has the potential for the rapid and on-site analyses of p-aminophenol in environmental water and human urine samples.

  7. Ionic liquid-based electromembrane extraction and its comparison with traditional organic solvent based electromembrane extraction for the determination of strychnine and brucine in human urine.

    Science.gov (United States)

    Sun, Jian-Nan; Chen, Juan; Shi, Yan-Ping

    2014-07-25

    An ionic liquid-based electromembrane extraction (IL-EME) method was presented, and its performance was compared with 2-ethylnitrobenzene (ENB) based EME for the determination of strychnine and brucine in human urine. For the two methods, the fundamental extraction parameters such as supported liquid membrane, voltage, extraction time, pH values of sample solution and acceptor solution, temperature and salting-out effect were separately optimized. IL-EME provided 96- and 122-fold enrichment factors for strychnine and brucine, respectively, which were better than those obtained in EME (83- and 86-fold, respectively). The calibration curves were linear over the ranges of 20-720 μg L(-1) for strychnine and 20-640 μg L(-1) for brucine with the correlation coefficients higher than 0.9950. The repeatability of EME and IL-EME were evaluated by five parallel experiments giving the relative standard deviations of 5.12-6.98%. As the results indicated, compared with ENB based EME, the proposed IL-EME is more reliable and could provide better extraction performance for the determination of strychnine and brucine in human urine. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Solidified floating organic drop microextraction combined with high performance liquid chromatography for the determination of carbamazepine in human plasma and urine samples

    Institute of Scientific and Technical Information of China (English)

    Mohammad ASADI; Ali Mohammad HAJI SHABANI; Shayessteh DADFARNIA; Bijan ABBASI

    2015-01-01

    Solidified floating organic drop microextraction( SFODME)in combination with high performance liquid chromatography was used for separation/preconcentration and determination of carbamazepine( CBZ)in human plasma and urine samples. Parameters that affect the extraction efficiency such as the type and volume of extraction solvent,ionic strength,sodium hydroxide concentration,stirring rate,sample volume and extrac-tion time,were investigated and optimized. Under the optimum conditions( extraction solvent,40 μL of 1-un-decanol;sodium hydroxide concentration,1 mol/L;temperature,50 ℃;stirring speed,400 r/min;sample vol-ume,8 mL;sodium chloride concentration,3%( w/v)and extraction time,60 min)the calibration curve was found to be linear in the mass concentration range of 0. 4-700. 0 μg/L. The limit of detection( LOD)was 0. 1μg/L and the relative standard deviation( RSD)for six replicate extraction and determination of carbamazepine at 100 μg/L level was found to be 4. 1%. The method was successfully applied to the determination of CBZ in human plasma and urine samples.

  9. Trace analysis of doxylamine succinate in animal feed, human urine, and wastewater by GC using a rubidium-sensitized nitrogen detector

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, H.C. Jr.; Holder, C.L.; Bowman, M.C.

    1982-08-01

    Doxylamine succinate, a drug used as a sleep-inducing agent, an antihistamine, and in a therapeutic formulation taken by pregnant women as an antinauseant, was scheduled for toxicological evaluation as part of a structure activity relationship study, with rats and mice, because a deficiency of such data exists with regard to many antihistamines. Analytical chemical procedures that ensure proper concentration, homogeneity, and stability of the drug in dosed feed, as well as the safety of personnel and the environment, were prerequisites for the toxicological tests. GC methods using a rubidium-sensitized nitrogen detector were developed for analysis of doxylamine succinate in animal feed, human urine, and wastewater at levels as low as 1 ppm, 100 ppb, and 100 ppb, respectively. Sample extracts were cleaned up by liquid-liquid partitioning, followed by additional cleanup on a column of silica gel. Data are presented concerning the stability of the drug in animal feed, extraction efficiencies, and the use of the silica gel cleanup column to separate the caffeine interference from doxylamine in extracts of human urine. Partition values and ancillary data concerning analysis of the drug in feed, by HPLC at levels as low as 10 ppm, are also reported.

  10. UHPLC-MS/MS method for the determination of bisphenol A and its chlorinated derivatives, bisphenol S, parabens, and benzophenones in human urine samples.

    Science.gov (United States)

    Vela-Soria, F; Ballesteros, O; Zafra-Gómez, A; Ballesteros, L; Navalón, A

    2014-06-01

    In the present work, a new method based on a sample treatment by dispersive liquid-liquid microextraction (DLLME) for the extraction of six bisphenols (bisphenol A, bisphenol S, and monochloro-, dichloro-, trichloro-, and tetrachlorobisphenol A), four parabens (methyl-, ethyl-, propyl-, and butylparaben), and six benzophenones (benzophenone-1, benzophenone-2, benzophenone-3, benzophenone-6, benzophenone-8, and 4-hydroxybenzophenone) in human urine samples, followed by ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis, is validated. An enzymatic treatment allows determining the total content of the target EDCs. The extraction parameters were accurately optimized using multivariate optimization strategies. Ethylparaben ring-(13)C6, benzophenone-d10, and bisphenol A-d16 were used as surrogates. Limits of quantification ranging from 0.1 to 0.6 ng mL(-1) and interday variabilities (evaluated as relative standard deviations) from 2.0 to 13.8% were obtained. The method was validated using matrix-matched standard calibration followed by a recovery assay with spiked samples. Recovery rates ranged from 94 to 106%. A good linearity, for concentrations up to 300 ng mL(-1) for parabens and 40 ng mL(-1) for benzophenones and bisphenols, was also obtained. The method was satisfactorily applied for the determination of target compounds in human urine samples from 20 randomly selected individuals.

  11. Electrochemical detection of toxic ractopamine and salbutamol in pig meat and human urine samples by using poly taurine/zirconia nanoparticles modified electrodes.

    Science.gov (United States)

    Rajkumar, Muniyandi; Li, Ying-Sheng; Chen, Shen-Ming

    2013-10-01

    Detection of ractopamine and salbutamol has been developed by employing the facile synthesis of poly taurine/zirconia nanoparticles (ZrO2) modified film glassy carbon electrode. The poly taurine/ZrO2 nanoparticles were directly utilized for the detection of ractopamine and salbutamol using linear sweep voltammetry (LSV). The modified electrode successfully shows the oxidation peak for ractopamine adsorption at 0.65V and salbutamol at 0.71V, which is purely based on the detection of adsorption signals of ractopamine and salbutamol, at the electrode surface. Furthermore, the electrochemical measurements and surface morphology were studied using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM) analysis. The modified electrode successfully detects the oxidation signals of ractopamine in the linear range of 1-28μM and salbutamol in the linear range of 5-220μM in laboratory samples. The proposed film also successfully detects the ractopamine signal (1-26μM) in pig meat samples and salbutamol signal (1-114μM) in human urine samples. It also exhibits two well-separated anodic oxidation peaks for uric acid and salbutamol in salbutamol-spiked human urine samples.

  12. Quantitation of organophosphorus nerve agent metabolites in human urine using isotope dilution gas chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Driskell, W Jack; Shih, Ming; Needham, Larry L; Barr, Dana B

    2002-01-01

    An isotope dilution gas chromatography-tandem mass spectrometric (GC-MS-MS) method was developed for quantitating the urinary metabolites of the organophosphorus nerve agents sarin, soman, tabun (GA), VX, and GF. Urine samples were concentrated by codistillation with acetonitrile, derivatized by methylation with diazomethane, and analyzed by GC-MS-MS. The limits of detection were less than 4 microg/L for all the analytes except for the GA metabolite, which had a limit of detection of less than 20 microg/L.

  13. The urine marker test

    DEFF Research Database (Denmark)

    Elbe, Anne-Marie; Jensen, Stine Nylandsted; Elsborg, Peter

    2016-01-01

    BACKGROUND: Urine sample collection for doping control tests is a key component of the World Anti-Doping Agency's fight against doping in sport. However, a substantial number of athletes experience difficulty when having to urinate under supervision. Furthermore, it cannot always be ensured...... that athletes are actually delivering their own urine. A method that can be used to alleviate the negative impact of a supervised urination procedure and which can also identify urine as coming from a specific athlete is the urine marker test. Monodisperse low molecular weight polyethylene glycols (PEGs......) are given orally prior to urination. Urine samples can be traced to the donor by analysis of the PEGs previously given. OBJECTIVE: The objective of this study was to investigate the use of the urine marker during urine doping control testing. METHODS: Two studies investigated athletes' acceptance...

  14. A competitive immunoassay for ultrasensitive detection of Hg{sup 2+} in water, human serum and urine samples using immunochromatographic test based on surface-enhanced Raman scattering

    Energy Technology Data Exchange (ETDEWEB)

    She, Pei; Chu, Yanxin [The Key Lab of Health Chemistry & Molecular Diagnosis of Suzhou, College of Chemistry, Chemical Engineering & Materials Science, Soochow University, Renai Road 199, Suzhou 215123 (China); Liu, Chunwei; Guo, Xun [OptoTrace (Suzhou) Technologies, Inc., STE 316, Building 4, No. 218, Xinghu Street, bioBAY, Suzhou Industrial Park, Suzhou 215123 (China); Zhao, Kang [The Key Lab of Health Chemistry & Molecular Diagnosis of Suzhou, College of Chemistry, Chemical Engineering & Materials Science, Soochow University, Renai Road 199, Suzhou 215123 (China); Li, Jianguo, E-mail: lijgsd@suda.edu.cn [The Key Lab of Health Chemistry & Molecular Diagnosis of Suzhou, College of Chemistry, Chemical Engineering & Materials Science, Soochow University, Renai Road 199, Suzhou 215123 (China); Du, Haijing; Zhang, Xiang [The Key Lab of Health Chemistry & Molecular Diagnosis of Suzhou, College of Chemistry, Chemical Engineering & Materials Science, Soochow University, Renai Road 199, Suzhou 215123 (China); Wang, Hong [OptoTrace (Suzhou) Technologies, Inc., STE 316, Building 4, No. 218, Xinghu Street, bioBAY, Suzhou Industrial Park, Suzhou 215123 (China); Deng, Anping, E-mail: denganping@suda.edu.cn [The Key Lab of Health Chemistry & Molecular Diagnosis of Suzhou, College of Chemistry, Chemical Engineering & Materials Science, Soochow University, Renai Road 199, Suzhou 215123 (China)

    2016-02-04

    An immunochromatographic test (ICT) strip was developed for ultrasensitive competitive immunoassay of Hg{sup 2+}. This strategy was achieved by combining the easy-operation and rapidity of ICT with the high sensitivity of surface-enhanced Raman scattering (SERS). Monoclonal antibody (mAb) against Hg{sup 2+} and Raman active substance 4-mercaptobenzoic acid (MBA) dual labelled gold nanoparticles (GNPs) were prepared as an immunoprobe. The Raman scattering intensity of MBA on the test line of the ICT strip was measured for quantitative determination of Hg{sup 2+}. The ICT was able to directly detect Hg{sup 2+} without complexing due to the specific recognition of the mAb with Hg{sup 2+}. The IC{sub 50} and limit of detection (LOD) of the assay for Hg{sup 2+} detection were 0.12 ng mL{sup −1} and 0.45 pg mL{sup −1}, respectively. There was no cross-reactivity (CR) of the assay with other nineteen ions and the ICT strips could be kept for 5 weeks without loss of activity. The recoveries of the assay for water, human serum and urine samples spiked with Hg{sup 2+} were in range of 88.3–107.3% with the relative standard deviations (RSD) of 1.5–9.5% (n = 3). The proposed ICT was used for the detection of Hg{sup 2+} in urine samples collected from Occupational Disease Hospital and the results were confirmed by cold-vapor atomic fluorescence spectroscopy (CV-AFS). The assay exhibited high sensitivity, selectivity, stability, precision and accuracy, demonstrating a promising method for the detection of trace amount of Hg{sup 2+} in environmental water samples and biological serum and urine samples. - Highlights: • The proposed ICT was able to directly detect Hg{sup 2+} without formation of Hg{sup 2+}-ligand complex. • The proposed ICT exhibited high sensitivity, specificity, stability, precision and accuracy for Hg{sup 2+} detection. • The proposed ICT was applicable for the detection of trace amount of Hg{sup 2+} in water, human serum and urine samples.

  15. Assessing Nitrogen and Phosphorus in Excreta from Grower-finisher Pigs Fed Prevalent Rations in Vietnam

    DEFF Research Database (Denmark)

    Van, Vu Thi Khanh; Sommer, Sven G.; Vu, C. C.

    2010-01-01

    Livestock production in Vietnam is, as in most Asian countries, increasing rapidly and changing into specialized highly intensified operations. The volume of animal excreta generated exceeds the capacity of the operation land base and cannot be utilized efficiently. As a consequence, there is a l......Livestock production in Vietnam is, as in most Asian countries, increasing rapidly and changing into specialized highly intensified operations. The volume of animal excreta generated exceeds the capacity of the operation land base and cannot be utilized efficiently. As a consequence......, there is a loss of plant nutrients from livestock farms that causes environmental pollution. This study carried out a feed and excretion experiment measuring fecal characteristic, daily fecal production, daily nitrogen and phosphorous excretion from grower-finisher pigs fed prevalent rations in Vietnam....... Furthermore, equations for assessing the excretion were tested, which can be used in farm models for optimal recycling of manure while focusing on reducing pollution. The results indicated that fecal production and nutrient excretion were affected by the different rations tested. This study showed that five...

  16. Co-digestion of swine excreta associated with increasing levels of crude glycerin

    Directory of Open Access Journals (Sweden)

    Marco Antonio Previdelli Orrico Junior

    2016-03-01

    Full Text Available ABSTRACT The objective of this research was to evaluate the performance of anaerobic co-digestion of swine excreta associated with increasing doses of crude glycerin and different hydraulic retention times (HRT. A completely randomized design was adopted in a 3 × 4 factorial arrangement composed of three HRT (10, 17, and 24 days and four crude glycerin doses (0, 5, 10, and 15 g/100 g of total solids [TS], with four replications per treatment. The assessed parameters were: biogas production potential, reductions of volatile solids (VS, chemical oxygen demand (COD, and most probable number of total and thermotolerant coliforms. The biogas production per added VS presented quadratic effect at 17 and 24 days of HRT, with ideal doses of 5.5 and 5.9 g of crude glycerin/100 g TS, respectively. There was no difference among glycerin doses at 10-day HRT for VS reductions; however, at HRT of 17 and 24 days, there were differences, with greater reduction of 61.1% for 5 g of crude glycerin/100 g TS at 24-day HRT. The COD reduction values showed an effect among retention times, in which the 24-day HRT provided the best results. Reductions in coliforms were greater than 99%, with no difference among treatments. Addition of 5 to 6 g of crude glycerin/100 g TS with a 24-day HRT is more effective in biogas production and reduction of VS, COD, and coliforms from co-digestion of swine excreta.

  17. Nest site selection and nutritional provision through excreta: a form of parental care in a tropical endogeic earthworm

    Directory of Open Access Journals (Sweden)

    Angel I. Ortiz-Ceballos

    2016-05-01

    Full Text Available Nest construction is a common form of parental care in soil organisms. However, it is unknown whether the tropical earthworm Pontoscolex corethrurus produces nests in soils with low nutritional quality habitats. Here we studied the reproductive behaviour and nest site selection of P. corethrurus, and tested the hypothesis whether P. corethrurus produces more cocoons in habitats with low nutritional quality. In bidimensional terrariums we evaluated the combined effect of the nutritional quality of habitat: (Poor Quality Habitat = PQH, Medium Quality Habitat = MQH, High Quality Habitat = HQH and soil depth (Shallow, Intermediate, Deep in a factorial 32 design. The number and biomass of cocoons, progeny and the production of internal and external excreta were evaluated. The quality habitat and depth of soil and their interaction had a significant effect on nest site construction and the deposition of internal excreta. Pontoscolex corethrurus built a higher amount of nests in the PQH-Intermediate and MQH-Intermediate treatments while more internal excreta were found in the HQH-Intermediate treatment. Offspring biomass was positively associated with internal excreta in the PQH (soil only and MQH (soil + grass treatments, suggesting that this could be a form of parental care. Since P. corethrurus produces more cocoons in low and medium quality habitats, while produces more internal excreta at high quality habitats, there does not seem to be an association between number of offspring and parental care. We suggest P. corethrurus could have two reproductive strategies that act as diversified bet-hedging (do not put all cocoons in one basket behavior in unpredictable environment, and thus build a higher amount of nests in low and medium quality habitats; and another where they produce more internal excreta as a form of parental care in high quality habitats. Parental care in the form of internal excreta may be particularly important in poor and medium

  18. Nest site selection and nutritional provision through excreta: a form of parental care in a tropical endogeic earthworm.

    Science.gov (United States)

    Ortiz-Ceballos, Angel I; Pérez-Staples, Diana; Pérez-Rodríguez, Paulino

    2016-01-01

    Nest construction is a common form of parental care in soil organisms. However, it is unknown whether the tropical earthworm Pontoscolex corethrurus produces nests in soils with low nutritional quality habitats. Here we studied the reproductive behaviour and nest site selection of P. corethrurus, and tested the hypothesis whether P. corethrurus produces more cocoons in habitats with low nutritional quality. In bidimensional terrariums we evaluated the combined effect of the nutritional quality of habitat: (Poor Quality Habitat = PQH, Medium Quality Habitat = MQH, High Quality Habitat = HQH) and soil depth (Shallow, Intermediate, Deep) in a factorial 3(2) design. The number and biomass of cocoons, progeny and the production of internal and external excreta were evaluated. The quality habitat and depth of soil and their interaction had a significant effect on nest site construction and the deposition of internal excreta. Pontoscolex corethrurus built a higher amount of nests in the PQH-Intermediate and MQH-Intermediate treatments while more internal excreta were found in the HQH-Intermediate treatment. Offspring biomass was positively associated with internal excreta in the PQH (soil only) and MQH (soil + grass) treatments, suggesting that this could be a form of parental care. Since P. corethrurus produces more cocoons in low and medium quality habitats, while produces more internal excreta at high quality habitats, there does not seem to be an association between number of offspring and parental care. We suggest P. corethrurus could have two reproductive strategies that act as diversified bet-hedging (do not put all cocoons in one basket) behavior in unpredictable environment, and thus build a higher amount of nests in low and medium quality habitats; and another where they produce more internal excreta as a form of parental care in high quality habitats. Parental care in the form of internal excreta may be particularly important in poor and medium quality habitats

  19. Evaluation of free water and water activity measurements as functional alternatives to total moisture content in broiler excreta and litter samples

    NARCIS (Netherlands)

    Hoeven-Hangoor, van der E.; Rademaker, C.; Paton, N.D.; Verstegen, M.W.A.; Hendriks, W.H.

    2014-01-01

    Litter moisture contents vary greatly between and within practical poultry barns. The current experiment was designed to measure the effects of 8 different dietary characteristics on litter and excreta moisture content. Additionally, free water content and water activity of the excreta and litter

  20. The ferric yersiniabactin uptake receptor FyuA is required for efficient biofilm formation by urinary tract infectious Escherichia coli in human urine

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Ferrieres, Lionel; Klemm, Per

    2008-01-01

    Urinary tract infection (UTI) is the most common infection in patients with indwelling urinary catheters, and bacterial biofilm formation is a major problem in this type of infection. Escherichia coli is responsible for the large majority of UTIs. Free iron is strictly limited in the human urinary...... tract and there is fierce competition between the host and infectious bacteria for this essential metal. Urinary tract infectious E coli have highly efficient mechanisms of iron acquisition, one of which is the yersiniabactin system. The fyuA gene, encoding the yersiniabactin receptor, is one...... of the most upregulated genes in biofilm; it was upregulated 63-fold in the E coli UTI strain VR50. FyuA was found to be highly important for biofilm formation in iron-poor environments such as human urine. Mutants in fyuA show aberrant biofilm formation and the cells become filamentous; a VR50fyuA mutant...

  1. A competitive immunoassay for ultrasensitive detection of Hg(2+) in water, human serum and urine samples using immunochromatographic test based on surface-enhanced Raman scattering.

    Science.gov (United States)

    She, Pei; Chu, Yanxin; Liu, Chunwei; Guo, Xun; Zhao, Kang; Li, Jianguo; Du, Haijing; Zhang, Xiang; Wang, Hong; Deng, Anping

    2016-02-01

    An immunochromatographic test (ICT) strip was developed for ultrasensitive competitive immunoassay of Hg(2+). This strategy was achieved by combining the easy-operation and rapidity of ICT with the high sensitivity of surface-enhanced Raman scattering (SERS). Monoclonal antibody (mAb) against Hg(2+) and Raman active substance 4-mercaptobenzoic acid (MBA) dual labelled gold nanoparticles (GNPs) were prepared as an immunoprobe. The Raman scattering intensity of MBA on the test line of the ICT strip was measured for quantitative determination of Hg(2+). The ICT was able to directly detect Hg(2+) without complexing due to the specific recognition of the mAb with Hg(2+). The IC50 and limit of detection (LOD) of the assay for Hg(2+) detection were 0.12 ng mL(-1) and 0.45 pg mL(-1), respectively. There was no cross-reactivity (CR) of the assay with other nineteen ions and the ICT strips could be kept for 5 weeks without loss of activity. The recoveries of the assay for water, human serum and urine samples spiked with Hg(2+) were in range of 88.3-107.3% with the relative standard deviations (RSD) of 1.5-9.5% (n = 3). The proposed ICT was used for the detection of Hg(2+) in urine samples collected from Occupational Disease Hospital and the results were confirmed by cold-vapor atomic fluorescence spectroscopy (CV-AFS). The assay exhibited high sensitivity, selectivity, stability, precision and accuracy, demonstrating a promising method for the detection of trace amount of Hg(2+) in environmental water samples and biological serum and urine samples.

  2. Kinetics and modeling of sulfonamide antibiotic degradation in wastewater and human urine by UV/H2O2 and UV/PDS.

    Science.gov (United States)

    Zhang, Ruochun; Yang, Yongkui; Huang, Ching-Hua; Zhao, Lin; Sun, Peizhe

    2016-10-15

    Sulfonamide antibiotics have been frequently detected in the aquatic environment and are of emerging concern due to their adverse bio-effect and potential of inducing antibiotic resistance. This study investigated the degradation kinetics of sulfonamide antibiotics in synthetic wastewater and hydrolyzed human urine by low pressure (LP) UV, UV/H2O2 and UV/peroxydisulfate (PDS). Direct photolysis rates of sulfonamide antibiotics varied and depended on the structures. Sulfonamides with a five-membered heterocyclic group underwent faster direct photolysis. For indirect photolysis processes, second-order rate constants of sulfonamide antibiotics with hydroxyl radical, sulfate radical and carbonate radical were determined, which were (6.21-9.26) × 10(9), (0.77-16.1) × 10(10) and (1.25-8.71) × 10(8) M(-1) s(-1), respectively. A dynamic model was applied and successfully predicted the degradation kinetics of sulfonamides in different water matrices. In synthetic wastewater, carbonate radical contributed to approximately 10% of the overall removal, whereas in synthetic hydrolyzed urine, carbonate radical was the dominant reactive species to degrade sulfonamides. Sulfonamide antibiotics were eliminated more efficiently in synthetic hydrolyzed urine than in synthetic wastewater and UV/PDS was more efficient than UV/H2O2 to degrade most sulfonamides. Energy evaluation showed that UV/PDS costs less energy than LPUV and UV/H2O2 under the experimental conditions applied in this study, particularly for sulfonamides whose indirect photolysis overweighed direct photolysis. By varying UV dose and oxidant dose, the UV/H2O2 process can be optimized to achieve higher efficiency than the UV/PDS process in synthetic wastewater.

  3. Determination of risedronate in human urine by column-switching ion-pair high-performance liquid chromatography with ultraviolet detection.

    Science.gov (United States)

    Vallano, P T; Shugarts, S B; Kline, W F; Woolf, E J; Matuszewski, B K

    2003-08-25

    An HPLC assay for the determination of risedronate in human urine was developed and validated. Risedronate and the internal standard were isolated from 5-ml urine samples in a two-part procedure. First, the analytes were precipitated from urine along with endogenous phosphates as calcium salts by the addition of CaCl(2) at alkaline pH. The precipitate was then dissolved in 0.05 M ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid and subjected to ion-pair solid-phase extraction using a Waters HLB cartridge (1 ml, 30 mg) with 1-octyltriethylammonium phosphate as the ion-pair reagent. Following extraction, the analytes were initially separated from the majority of co-extracted endogenous components on a Waters X-Terra RP18 (4.6 x 50 mm, 3.5 microm) column. The effluent from the X-Terra was "heart-cut" onto a Phenomenex Synergi Polar RP (4.6 x 150 mm, 4 microm) column for final separation. UV detection (lambda=262 nm) was used to quantitate risedronate in the concentration range of 7.5-250 ng/ml. Mean recovery was 83.3% for risedronate and 86.5% for the internal standard. The intra-day precision of the assay, as assessed by replicate (n=5) standard curves, was better than 6% RSD for all points on the standard curve. Within-day accuracy for the standards ranged from 96.3 to 106.1% of nominal. Inter-day precision for quality controls assayed over a 3-week period was better than 5%, while inter-day accuracy was within 90% of nominal. The assay was employed to analyze samples collected during a clinical pharmacokinetics study.

  4. Vanadium in foods and in human body fluids and tissues.

    Science.gov (United States)

    Byrne, A R; Kosta, L

    1978-07-01

    Using neutron activation analysis, vanadium was analysed in a range of foods, human body fluids and tissues. On the basis of these results and those of other workers, it was concluded that daily dietary intake amounts to some tens of micrograms. Analysis of body fluids (including milk, blood and excreta) and organs and tissues provided an estimate for the total body pool of vanadium in man of about 100 microgram. Vanadium was not detectable in blood and urine at the level of 0.3 ng/g, while low levels were found in muscle, fat, bone, teeth and other tissues. The relationship between dietary intake to pulmonary absorption is discussed in relation to the occurrence of vanadium in man-made air particulates. The very low levels found in milks and eggs suggest minimal vanadium requirements in growth. The findings are discussed in the light of previous results and also in relation to the possible essentiality of vanadium.

  5. Simultaneous Separation of Eight Benzodiazepines in Human Urine Using Field-Amplified Sample Stacking Micellar Electrokinetic Chromatography.

    Science.gov (United States)

    Oledzka, Ilona; Kulińska, Zofia; Prahl, Adam; Baczek, Tomasz

    2015-01-01

    A novel approach for the simultaneous quantification of eight benzodiazepines (BZDs) using dispersive liquid-liquid microextraction (DLLME) and field-amplified sample stacking (FASS) combined with micellar electrokinetic chromatography (MEKC) was investigated and evaluated in the context of precision, accuracy, sensitivity, linearity, detection and limits of quantification (LOQ). The absolute recovery rates of BZDs were above 90.65%. The limits of detection (LOD) were 20 ng/mL for chlordiazepoxide, estazolam, temazepam and midazolam, and 30 ng/mL for clonazepam, lorazepam, lormetazepam and medazepam, while the LOQ was set at 50 ng/mL for chlordiazepoxide, estazolam, temazepam and midazolam, and 100 ng/mL for clonazepam, lorazepam, lormetazepam and medazepam. Linearity was confirmed in the range of 50-2,000 ng/mL for chlordiazepoxide, estazolam, temazepam and midazolam, and 100-2,000 ng/mL for clonazepam, lorazepam, lormetazepam and medazepam, with a correlation coefficient greater than 0.9987 for all analytes. The elaborated procedure meets all the requirements of analytical methods. During the extraction procedure, a mixture of 1 mL of ethanol and 500 µL of dichloromethane, used as the disperser and extraction solvent, respectively, was rapidly injected into 3 mL of a urine sample. A significant improvement in sensitivity was achieved when DLLME was used to extract BZDs from the urine sample and FASS as an on-line preconcentration technique was developed. For the best separation of analytes, the running buffer was composed of 30 mM SDS, 10 mM sodium tetraborate and 15% methanol (pH 8.8), whereas a sample buffer was composed of 10 mM SDS and 2 mM sodium tetraborate. Moreover, a fused-silica capillary [inner diameter (i.d.) of 75 µm and length of 50 cm], photodiode array detection, pneumatic injection for 15 s and a voltage of 23 kV were applied. The applicability of the method has been confirmed for the analysis of BZD in urine samples collected from patients who

  6. Development of an isotope labeling ultra-high performance liquid chromatography mass spectrometric method for quantification of acylglycines in human urine

    Energy Technology Data Exchange (ETDEWEB)

    Stanislaus, Avalyn; Guo, Kevin [Department of Chemistry, University of Alberta, Edmonton, Alberta (Canada); Li Liang, E-mail: Liang.Li@ualberta.ca [Department of Chemistry, University of Alberta, Edmonton, Alberta (Canada)

    2012-10-31

    Graphical abstract: - Abstract: Acylglycines play a crucial regulatory and detoxification role in the accumulation of the corresponding acyl CoA esters and are an important class of metabolites in the diagnoses of inborn errors of metabolism. Sensitive quantification of a large number of acylglycines not only improves diagnosis but also enables the discovery of potential new biomarkers of diseases. We report an ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS) method for quantifying acylglycines in human urine with high sensitivity. This method is based on the use of a newly developed isotope labeling reagent, p-dimethylaminophenacyl (DmPA) bromide, to label acylglycines to improve detection sensitivity. Eighteen acylglycines, namely acetylglycine, propionylglycine, isobutyrylglycine, butyrylglycine, 4-hydroxyphenylacetylglycine, 2-furoylglycine, tiglylglycine, 2-methybutyrylglycine, 3-methylcrotonylglycine, isovalerylglycine, valerylglycine, hexanoylglycine, phenylacetylglycine, phenylpropionylglycine, glutarylglycine, heptanoylglycine, octanoylglycine and suberylglycine, were measured. This method uses calibration standards prepared in surrogate matrix (un-derivatized urine) and stable-isotope labeled analytes as the internal standards. The analysis was carried out in the positive ion detection mode using multiple reaction monitoring (MRM) survey scans. The calibration curves were validated over the range of 1.0-500 nM. The method achieved a lower limit of quantitation (LLOQ) of 1-5 nM for all analytes, as measured by the standard derivations associated with calibration curves and confirmed in surrogate matrix; the signal-to-noise ratio at LLOQ ranged from 12.50 to 156.70. Both accuracy (% RE or relative error) and precision (% CV) were <15%. Matrix effects were minimized using the surrogate matrix. All eighteen analytes were stable in urine for at least 5 h at room temperature, autosampler (4 Degree-Sign C) for 24 h, 7 weeks at -20

  7. A pitfall in diagnosis of human prion diseases using detection of protease-resistant prion protein in urine. Contamination with bacterial outer membrane proteins.

    Science.gov (United States)

    Furukawa, Hisako; Doh-ura, Katsumi; Okuwaki, Ryo; Shirabe, Susumu; Yamamoto, Kazuo; Udono, Heiichiro; Ito, Takashi; Katamine, Shigeru; Niwa, Masami

    2004-05-28

    Because a definite diagnosis of prion diseases relies on the detection of the abnormal isoform of prion protein (PrPSc), it has been urgently necessary to establish a non-invasive diagnostic test to detect PrPSc in human prion diseases. To evaluate diagnostic usefulness and reliability of the detection of protease-resistant prion protein in urine, we extensively analyzed proteinase K (PK)-resistant proteins in patients affected with prion diseases and control subjects by Western blot, a coupled liquid chromatography and mass spectrometry analysis, and N-terminal sequence analysis. The PK-resistant signal migrating around 32 kDa previously reported by Shaked et al. (Shaked, G. M., Shaked, Y., Kariv-Inbal, Z., Halimi, M., Avraham, I., and Gabizon, R. (2001) J. Biol. Chem. 276, 31479-31482) was not observed in this study. Instead, discrete protein bands with an apparent molecular mass of approximately 37 kDa were detected in the urine of many patients affected with prion diseases and two diseased controls. Although these proteins also gave strong signals in the Western blot using a variety of anti-PrP antibodies as a primary antibody, we found that the signals were still detectable by incubation of secondary antibodies alone, i.e. in the absence of the primary anti-PrP antibodies. Mass spectrometry and N-terminal protein sequencing analysis revealed that the majority of the PK-resistant 37-kDa proteins in the urine of patients were outer membrane proteins (OMPs) of the Enterobacterial species. OMPs isolated from these bacteria were resistant to PK and the PK-resistant OMPs from the Enterobacterial species migrated around 37 kDa on SDS-PAGE. Furthermore, nonspecific binding of OMPs to antibodies could be mistaken for PrPSc. These findings caution that bacterial contamination can affect the immunological detection of prion protein. Therefore, the presence of Enterobacterial species should be excluded in the immunological tests for PrPSc in clinical samples, in

  8. Online micro-solid-phase extraction based on boronate affinity monolithic column coupled with high-performance liquid chromatography for the determination of monoamine neurotransmitters in human urine.

    Science.gov (United States)

    Yang, Xiaoting; Hu, Yufei; Li, Gongke

    2014-05-16

    Quantification of monoamine neurotransmitters is very important in diagnosing and monitoring of patients with neurological disorders. We developed an online analytical method to selectively determine urinary monoamine neurotransmitters, which coupled the boronate affinity monolithic column micro-solid-phase extraction with high-performance liquid chromatography (HPLC). The boronate affinity monolithic column was prepared by in situ polymerization of vinylphenylboronic acid (VPBA) and N,N'-methylenebisacrylamide (MBAA) in a stainless capillary column. The prepared monolithic column showed good permeability, high extraction selectivity and capacity. The column-to-column reproducibility was satisfactory and the enrichment factors were 17-243 for four monoamine neurotransmitters. Parameters that influence the online extraction efficiency, including pH of sample solution, flow rate of extraction and desorption, extraction volume and desorption volume were investigated. Under the optimized conditions, the developed method exhibited low limit of detection (0.06-0.80μg/L), good linearity (with R(2) between 0.9979 and 0.9993). The recoveries in urine samples were 81.0-105.5% for four monoamine neurotransmitters with intra- and inter-day RSDs of 2.1-8.2% and 3.7-10.6%, respectively. The online analytical method was sensitive, accurate, selective, reliable and applicable to analysis of trace monoamine neurotransmitters in human urine sample.

  9. The Determination of Nitrate and Nitrite in Human Urine and Blood by High-Performance Liquid Chromatography and Cloud-Point Extraction.

    Science.gov (United States)

    Zhao, Jiao; Wang, Jun; Yang, Yaling; Lu, Yunhui

    2015-08-01

    A simple efficient and practical separation/preconcentration coupled with HPLC method for the determination nitrate and low concentrations of nitrite in human urine and blood was investigated. The method is based on precolumn derivatization using the Griess reaction and cloud-point extraction (CPE) of nitrite anion and direct determination of nitrate using its UV absorbance by ion-pair HPLC. The chromatographic process with detection at two wavelengths (510 and 220 nm) allows the determination of nitrite and nitrate. Decolorization and protein precipitation of urine and blood was applied to overcome the interference of matrix and enhance the sensitivity. The method was validated for linearity, accuracy and precision. Under the optimum conditions, the linear range of nitrite from 10 to 1,000 ng/mL and nitrate from 0.1 to 10 µg/mL. Product recoveries ranged from 92.4 to 99.9%. The limits of detection were 1 ng/mL and 0.1 µg/mL for nitrite and nitrate, respectively. Therefore, the technique was simple and reliable, with potential application in biological sample analysis of nitrate and nitrite.

  10. Determination of free and total bisphenol A in human urine to assess daily uptake as a basis for a valid risk assessment.

    Science.gov (United States)

    Völkel, Wolfgang; Kiranoglu, Mandy; Fromme, Hermann

    2008-07-10

    Bisphenol A (BPA) is widely distributed and exhibits weak estrogenic activity. In contrast to BPA, the corresponding glucuronide metabolite is not estrogenic. Therefore, free and total BPA were determined in human urine samples to assess the significance of free BPA for risk assessment. In only 10% of 474 samples from 287 subjects was free BPA detected in a range from LOD (0.3 microg/l) to 2.5 microg/l. Due to sample contamination with low amounts ( approximately 1 microg/l) of BPA, house dust samples were independently collected in homes but not from persons who provide urine samples and analysed for BPA to check for potential sources of contamination. BPA was found in the range from 117 to 1 486 microg/kg (median: 553 microg/kg) dust. Additionally, BPA and d(16)-BPA were administered to a volunteer to demonstrate the problem of contamination. In comparison to low levels of free BPA (LOD and 9.3 microg/l.

  11. Ultrasound-assisted emulsification microextraction for the determination of ephedrines in human urine by capillary electrophoresis with direct injection. Comparison with dispersive liquid-liquid microextraction.

    Science.gov (United States)

    Alshana, Usama; Göğer, Nilgün G; Ertaş, Nusret

    2012-08-01

    Ultrasound-assisted emulsification microextraction and dispersive liquid-liquid microextraction were compared for extraction of ephedrine, norephedrine, and pseudoephedrine from human urine samples prior to their determination by capillary electrophoresis. Formation of a microemulsion of the organic extract with an aqueous solution (at pH 3.2) containing 10% methanol facilitated the direct injection of the final extract into the capillary. Influential parameters affecting extraction efficiency were systematically studied and optimized. In order to enhance the sensitivity further, field-amplified sample injection was applied. Under optimum extraction and stacking conditions, enrichment factors of up to 140 and 1750 as compared to conventional capillary zone electrophoresis were obtained resulting in limits of detection of 12-33 μg/L and 1.0-2.8 μg/L with dispersive liquid-liquid microextraction and ultrasound-assisted emulsification microextraction when combined with field-amplified sample injection. Calibration graphs showed good linearity for urine samples by both methods with coefficients of determination higher than 0.9973 and percent relative standard deviations of the analyses in the range of 3.4-8.2% for (n = 5). The results showed that the use of ultrasound to assist microextraction provided higher extraction efficiencies than disperser solvents, regarding the hydrophilic nature of the investigated analytes.

  12. Effect of magnetic field strength on NMR-based metabonomic human urine data. Comparative study of 250, 400, 500, and 800 MHz.

    Science.gov (United States)

    Bertram, Hanne Christine; Malmendal, Anders; Petersen, Bent O; Madsen, Jens Chr; Pedersen, Henrik; Nielsen, Niels Chr; Hoppe, Camilla; Mølgaard, Christian; Michaelsen, Kim F; Duus, Jens Ø

    2007-09-15

    Metabonomic analysis of urine utilizing high-resolution NMR spectroscopy and chemometric techniques has proven valuable in characterizing the biochemical response to an intervention. To assess the effect of magnetic field strength on information contained in NMR-based metabonomic data sets, 1H NMR spectra were acquired on 250-, 400-, 500-, and 800-MHz instruments, respectively, on the same set of human urine samples collected before and after dietary interventions with milk and with meat proteins. Partial least-squares regression discriminant analyses (PLS-DA) were performed in order to elucidate the ability of the 1H spectra acquired at various field strengths to identify possible spectral differences and discriminate between pre- and postintervention samples. The loadings from PLS-DA contained the same spectral regions, implying that the same metabolites were involved in the discrimination independent of magnetic field strength. The investigation revealed a strong increase in prediction performance and thereby spectral information content when increasing the magnetic field strength from 250 to 500 MHz, while from 500 to 800 MHz the increase was less pronounced.

  13. Development of a novel polystyrene/metal-organic framework-199 electrospun nanofiber adsorbent for thin film microextraction of aldehydes in human urine.

    Science.gov (United States)

    Liu, Feilong; Xu, Hui

    2017-01-01

    In this work, electrospun polystyrene/metal-organic frameworks-199 (PS/MOF-199) nanofiber film was synthesized and investigated as a novel adsorbent for thin film microextraction (TFME) of aldehydes in human urine. Some properties of the prepared PS/MOF-199 nanofiber film, including morphology, structure, wettability, solvent stability and extraction performance were studied systematically. Porous fibrous structure, large surface area, good stability, strong hydrophobicity and excellent extraction efficiency were obtained for the film. Based on the PS/MOF-199 film, a thin film microextraction-high performance liquid chromatography (TFME-HPLC) method was developed, and the experimental parameters that affected the extraction and desorption were optimized. Under the optimal conditions, the limits of detection (LODs) were in the range of 4.2-17.3nmolL(-1) for the analysis of six aldehydes. Good linearity was achieved with correlation coefficients (R(2)) being lager than 0.9943. Satisfactory recovery (82-112%) and acceptable reproducibility (relative standard deviation: 2.1-13.3%) were also obtained for the method. The developed TFME-HPLC method has been successfully applied to the analysis of aldehyde metabolites in the urine samples of lung cancer patients and healthy people. The method possesses the advantages of simplicity, rapidity, cost-effective, sensitivity and non-invasion, it provides an alternative tool for the determination of aldehydes in complex sample matrices.

  14. Affinity-purified antibodies of defined specificity for use in a solid-phase microplate radioimmunoassay of human Tamm-Horsfall glycoprotein in urine.

    Science.gov (United States)

    Hunt, J S; McGiven, A R; Groufsky, A; Lynn, K L; Taylor, M C

    1985-05-01

    Rabbit antibodies to human Tamm-Horsfall glycoprotein (prepared by salt precipitation from normal urine) were purified by affinity chromatography using columns containing Tamm-Horsfall glycoprotein linked to CNBr-activated Sepharose 4B. The specificity of these antibodies was determined by analysis of their binding characteristics on Western blots of Tamm-Horsfall protein from sodium dodecyl sulphate/polyacrylamide gradient gels and comparison with the reactivity of monoclonal antibodies to this glycoprotein. Optimal conditions of adsorption to poly(vinyl chloride) microtitre plates were established such that these purified antibodies could be used in a solid-phase radioimmunoassay for the determination of urinary Tamm-Horsfall-glycoprotein concentration. The specificity of the immunoassay was confirmed by competitive inhibition of the urinary Tamm-Horsfall glycoprotein by purified freeze-dried material in solution. A standard curve obtained with this material showed the radioimmunoassay to have a sensitivity of at least 5 ng/ml, with linearity between 30 and 600 ng/ml. The mean coefficient of variation over the linear section of the curve was 11.3 +/- 2.2% (n = 13). The effects of dialysis and freezing of urine samples before determination of Tamm-Horsfall-glycoprotein concentrations were investigated and the mean 24 h urinary excretion rate in 60 normal donors was shown to be 84.9 +/- 44.1 mg.

  15. Simultaneous extraction and quantification of lamotrigine, phenobarbital, and phenytoin in human plasma and urine samples using solidified floating organic drop microextraction and high-performance liquid chromatography.

    Science.gov (United States)

    Asadi, Mohammad; Dadfarnia, Shayessteh; Haji Shabani, Ali Mohammad; Abbasi, Bijan

    2015-07-01

    A novel and simple method based on solidified floating organic drop microextraction followed by high-performance liquid chromatography with ultraviolet detection has been developed for simultaneous preconcentration and determination of phenobarbital, lamotrigine, and phenytoin in human plasma and urine samples. Factors affecting microextraction efficiency such as the type and volume of the extraction solvent, sample pH, extraction time, stirring rate, extraction temperature, ionic strength, and sample volume were optimized. Under the optimum conditions (i.e. extraction solvent, 1-undecanol (40 μL); sample pH, 8.0; temperature, 25°C; stirring rate, 500 rpm; sample volume, 7 mL; potassium chloride concentration, 5% and extraction time, 50 min), the limits of detection for phenobarbital, lamotrigine, and phenytoin were 1.0, 0.1, and 0.3 μg/L, respectively. Also, the calibration curves for phenobarbital, lamotrigine, and phenytoin were linear in the concentration range of 2.0-300.0, 0.3-200.0, and 1.0-200.0 μg/L, respectively. The relative standard deviations for six replicate extractions and determinations of phenobarbital, lamotrigine, and phenytoin at 50 μg/L level were less than 4.6%. The method was successfully applied to determine phenobarbital, lamotrigine, and phenytoin in plasma and urine samples. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Simultaneous determination of six toxic alkaloids in human plasma and urine using capillary zone electrophoresis coupled to time-of-flight mass spectrometry.

    Science.gov (United States)

    Yu, Zhuhong; Wu, Zhongping; Gong, Feijun; Wong, Rong; Liang, Chen; Zhang, Yurong; Yu, Yunqiu

    2012-10-01

    A novel capillary zone electrophoresis separation coupled to electro spray ionization time-of-flight mass spectrometry method was developed for the simultaneous analysis of six toxic alkaloids: brucine, strychnine, atropine sulfate, anisodamine hydrobromide, scopolamine hydrobromide and anisodine hydrobromide in human plasma and urine. To obtain optimal sensitivity, a solid-phase extraction method using Oasis MCX cartridges (1 mL, 30 mg; Waters, USA) for the pretreatment of samples was used. All compounds were separated by capillary zone electrophoresis at 25 kV within 12 min in an uncoated fused-silica capillary of 75 μm id × 100 cm and were detected by time-of-flight mass spectrometry. This method was validated with regard to precision, accuracy, sensitivity, linear range, limit of detection (LOD), and limit of quantification (LOQ). In the plasma and urine samples, the linear calibration curves were obtained over the range of 0.50-100 ng/mL. The LOD and LOQ were 0.2-0.5 ng/mL and 0.5-1.0 ng/mL, respectively. The intra- and interday precision was better than 12% and 13%, respectively. Electrophoretic peaks could be identified by mass analysis.

  17. Semi-automated solid phase extraction method for the mass spectrometric quantification of 12 specific metabolites of organophosphorus pesticides, synthetic pyrethroids, and select herbicides in human urine.

    Science.gov (United States)

    Davis, Mark D; Wade, Erin L; Restrepo, Paula R; Roman-Esteva, William; Bravo, Roberto; Kuklenyik, Peter; Calafat, Antonia M

    2013-06-15

    Organophosphate and pyrethroid insecticides and phenoxyacetic acid herbicides represent important classes of pesticides applied in commercial and residential settings. Interest in assessing the extent of human exposure to these pesticides exists because of their widespread use and their potential adverse health effects. An analytical method for measuring 12 biomarkers of several of these pesticides in urine has been developed. The target analytes were extracted from one milliliter of urine by a semi-automated solid phase extraction technique, separated from each other and from other urinary biomolecules by reversed-phase high performance liquid chromatography, and detected using tandem mass spectrometry with isotope dilution quantitation. This method can be used to measure all the target analytes in one injection with similar repeatability and detection limits of previous methods which required more than one injection. Each step of the procedure was optimized to produce a robust, reproducible, accurate, precise and efficient method. The required selectivity and sensitivity for trace-level analysis (e.g., limits of detection below 0.5ng/mL) was achieved using a narrow diameter analytical column, higher than unit mass resolution for certain analytes, and stable isotope labeled internal standards. The method was applied to the analysis of 55 samples collected from adult anonymous donors with no known exposure to the target pesticides. This efficient and cost-effective method is adequate to handle the large number of samples required for national biomonitoring surveys. Published by Elsevier B.V.

  18. Association between aflatoxin M1 excreted in human urine samples with the consumption of milk and dairy products.

    Science.gov (United States)

    Mohd Redzwan, Sabran; Rosita, Jamaluddin; Mohd Sokhini, Abdul Mutalib; Nurul Aqilah, Abdul Rahman

    2012-12-01

    This study aimed to find the association between urinary aflatoxin M(1) level and milk and dairy products consumption. Of 160 morning urine samples collected, aflatoxin M(1) was detected in 61.3 % samples (n = 98) [mean ± SD = 0.0234 ± 0.0177 ng/mL; range = 0-0.0747 ng/mL]. Of these positive samples, 67.3 % (n = 66) had levels above the limit of detection. Respondents with intake of milk and dairy products above median (67.79 g/day) had significantly high level of AFM(1) compared to those with low intake. A significant and positive association (φ = 0.286) was found between milk and dairy products consumption and urinary aflatoxin M(1) level.

  19. Performance characteristics of DRI, CEDIA, and REMEDi systems for preliminary tests of amphetamines and opiates in human urine.

    Science.gov (United States)

    Huang, Min-Kun; Dai, Yu-Shan; Lee, Choung-Huei; Liu, Chiareiy; Tsay, Wen-Ing; Li, Jih-Heng

    2006-01-01

    Arrestee urine specimens (930) were tested with DRI, CEDIA, and REMEDi; those that tested positive for amphetamines and opiates (616 and 414, respectively) were then confirmed by gas chromatography-mass spectrometry. The performance characteristics of these three preliminary systems were evaluated using the following commonly used parameters: true positive, true negative, false positive, and false negative. The sensitivity, specificity, and efficiency of these methods were also calculated. Data derived from this study indicated DRI and CEDIA adapted by this study generated acceptable preliminary test results for amphetamine/methamphetamine and morphine/codeine, but not for MDA/MDMA and REMEDi has lower sensitivity than DRI and CEDIA, but with better specificity and efficiency, supporting its use under emergency room settings where drug concentrations in overdose cases are expectedly at high levels.

  20. Ocorrência de Cryptococcus neoformans em excretas de pombos na cidade de Pelotas, Estado do Rio Grande do Sul Occurrence of Cryptococcus neoformans in pigeon excrement in the city of Pelotas, State of Rio Grande do Sul

    Directory of Open Access Journals (Sweden)

    Renata Osório de Faria

    2010-04-01

    Full Text Available INTRODUÇÃO: O Cryptococcus neoformans é uma levedura capsulada, agente etiológico da criptococose em humanos e animais, encontrado em fontes ambientais, incluindo excretas de pombos, é uma importante causa de mortalidade em indivíduos imunodeprimidos em todo o mundo. MÉTODOS: Com o objetivo de verificar a ocorrência do Cryptococcus neoformans, em excretas de pombos, na Cidade de Pelotas, pesquisou-se 70 ambientes, incluindo prédios, praças e locais ao ar livre, da Cidade de Pelotas, RS. Após a coleta, os excrementos foram adicionados de salina com cloranfenicol, homogeneizados, semeados em ágar Sabouraud com cloranfenicol e ágar Níger e incubados a 32ºC. Para identificação, realizou-se exame direto, prova da fenoloxidase, urease, assimilação de carboidratos e cultura em meio CGB. RESULTADOS: Dos locais estudados (nº =70, em 26 (37,1% havia excretas de pombos. Estes lugares foram representados por prédios históricos (nº =8, torre de igreja (nº =1, engenhos e armazéns de arroz (nº =7, praça (nº =1 e locais ao ar livre (nº =9, o isolamento de Cryptococcus neoformans ocorreu em 26,9% (nº =7/26, destes locais. CONCLUSÕES: O estudo chama a atenção, para o isolamento do fungo em áreas urbanas, que apresentavam grande acúmulo de excretas, indicando um risco para a saúde pública, especialmente para indivíduos imunocomprometidos.INTRODUCTION: Cryptococcus neoformans is an encapsulated yeast and is the etiological agent for human and animal cryptococcosis. It is found in sources within the environment, including pigeon excrement, and is an important cause of mortality among immunocompromised individuals worldwide. METHODS: Seventy different environments in the city of Pelotas, Rio Grande do Sul, were surveyed for the purpose of investigating Cryptococcus neoformans occurrences in pigeon excreta. The environments included buildings, public squares and outdoor locations in the city. After collection, chloramphenicol

  1. Determination of trimethylamine, trimethylamine N-oxide, and taurine in human plasma and urine by UHPLC-MS/MS technique.

    Science.gov (United States)

    Awwad, Hussain Mohamad; Geisel, Juergen; Obeid, Rima

    2016-12-01

    Trimethylamine-N-oxide (TMAO) is produced in the liver from trimethylamine (TMA) and is an important cellular osmolyte and potential atherogenic factor. Taurine is involved in cholesterol metabolism and also serves as a cellular osmolyte. Given their significant biological functions, the development of reliable measurement techniques is crucial to further study their role in health and disease METHODS: A new ultrahigh performance liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous determination of TMA, TMAO, and taurine in plasma and urine. The method consisted of a deproteinization step using methanol/acetonitrile (15:85) that contained 0.2% formic acid and isotope-labeled internal standards. Samples were separated by centrifugation and injected into the UHPLC system. Quantification was conducted using a triple-quadrupole mass spectrometer detector with electrospray ionization interface in positive mode. The limits of detection ranged from 0.08 to 0.12μmol/L. The calibration curves were linear (r≥0.999) over the range examined (0.15-400μmol/L) for all compounds. The inter- and intra-day coefficients of variations were≤14.5% for TMA and ≤8% for TMAO and taurine. TMAO and taurine were found to be stable in EDTA plasma for at least 14 months at -70°C. Mean recoveries ranged from 95% to 109% and the relative matrix effects were≤4.0%. The method was applied to study physiological and pre-analytical factors in plasma and urine samples. The new UHPLC-MS/MS method has good accuracy, precision, and recovery. The assay combines simple sample processing with a short run time, making it well suited for high-throughput routine clinical or research purposes. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Rapid, automated online SPE-LC-QTRAP-MS/MS method for the simultaneous analysis of 14 phthalate metabolites and 5 bisphenol analogues in human urine.

    Science.gov (United States)

    Heffernan, A L; Thompson, K; Eaglesham, G; Vijayasarathy, S; Mueller, J F; Sly, P D; Gomez, M J

    2016-05-01

    Phthalates and bisphenol A (BPA) have received special attention in recent years due to their frequent use in consumer products and potential for adverse effects on human health. BPA is being replaced with a number of alternatives, including bisphenol S, bisphenol B, bisphenol F and bisphenol AF. These bisphenol analogues have similar potential for adverse health effects, but studies on human exposure are limited. Accurate measurement of multiple contaminants is important for estimating exposure. This paper describes a sensitive and automated method for the simultaneous determination of 14 phthalate metabolites, BPA and four bisphenol analogues in urine using online solid phase extraction coupled with high-performance liquid chromatography/tandem mass spectrometry using a hybrid triple-quadrupole linear ion trap mass spectrometer (LC-QTRAP-MS/MS), requiring very little sample volume (50µL). Quantification was performed under selected reaction monitoring (SRM) mode with negative electrospray ionization. The use of SRM combined with an enhanced product ion scan within the same analysis was examined. Unequivocal identification was provided by the acquisition of three SRM transitions per compound and isotope dilution. The analytical performance of the method was evaluated in synthetic and human urine. Linearity of response over three orders of magnitude was demonstrated for all of the compounds (R(2)>0.99), with method detection limits of 0.01-0.5ng/mL and limits of reporting of 0.07-3.1ng/mL. Accuracy ranged from 93% to 113% and inter- and intra-day precision were bisphenols, with median concentrations ranging from 0.3ng/mL (bisphenol S) to 18.5ng/mL (monoethyl phthalate).

  3. Evaluation of uv spectrophotometry for estimation of nitrite and nitrate in nitrified urine

    OpenAIRE

    Santos, Ana Teresa Lourenço

    2014-01-01

    Water is a limited resource for which demand is growing. Contaminated water from inadequate wastewater treatment provides one of the greatest health challenges as it restricts development and increases poverty in emerging and developing countries. Therefore, the connection between wastewater and human health is linked to access to sanitation and to human waste disposal. Adequate sanitation is expected to create a barrier between disposed human excreta and sources of drinking water. Different ...

  4. Urine specific gravity test

    Science.gov (United States)

    ... medlineplus.gov/ency/article/003587.htm Urine specific gravity test To use the sharing features on this page, please enable JavaScript. Urine specific gravity is a laboratory test that shows the concentration ...

  5. Protein urine test

    Science.gov (United States)

    ... Urine albumin; Proteinuria; Albuminuria Images White nail syndrome Protein urine test References Gerber GS, Brendler CB. Evaluation of the urologic patient: history, physical examination, and urinalysis. In: Wein AJ, Kavoussi ...

  6. Urine drug screen

    Science.gov (United States)

    Drug screen -- urine ... detect the presence of illegal and some prescription drugs in your urine. Their presence indicates that you recently used these drugs. Some drugs may remain in your system for ...

  7. Urine concentration test

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/article/003608.htm Urine concentration test To use the sharing features on this page, please enable JavaScript. A urine concentration test measures the ability of the kidneys to ...

  8. A multiclass method for the analysis of endocrine disrupting chemicals in human urine samples. Sample treatment by dispersive liquid-liquid microextraction.

    Science.gov (United States)

    Vela-Soria, F; Ballesteros, O; Zafra-Gómez, A; Ballesteros, L; Navalón, A

    2014-11-01

    The population is continuously exposed to endocrine disrupting chemicals (EDCs). This has influenced an increase in diseases and syndromes that are more frequent nowadays. Therefore, it is necessary to develop new analytical procedures to evaluate the exposure with the ultimate objective of establishing, in an accurate way, relationships between EDCs and harmful health effects. In the present work, a new method based on a sample treatment by dispersive liquid-liquid microextraction (DLLME) for the extraction of six parabens (methyl-, ethyl-, isopropyl-, propyl-, isobutyl and butylparaben), six benzophenones (benzophenone-1, benzophenone-2, benzophenone-3, benzophenone-6, benzophenone-8 and 4-hydroxybenzophenone) and two bisphenols (bisphenol A and bisphenol S) in human urine samples, followed by gas chromatography-tandem mass spectrometry (GC-MS/MS) analysis is proposed. An enzymatic treatment allows determining the total content of the target EDCs. The extraction parameters were accurately optimized using multivariate optimization strategies. Ethylparaben ring-(13)C6 and bisphenol A-d16 were used as surrogates. Found limits of quantification ranging from 0.2 to 0.5 ng mL(-1) and inter-day variability (evaluated as relative standard deviation) ranging from 2.0% to 14.9%. The method was validated using matrix-matched standard calibration followed by a recovery assay with spiked samples. Recovery rates ranged from 94% to 105%. A good linearity, for concentrations up to 300 ng mL(-1) for parabens and 40 ng mL(-1) for benzophenones and bisphenols, respectively, was obtained. The method was satisfactorily applied for the determination of target compounds in human urine samples from 20 randomly selected individuals.

  9. Cold column trapping-cloud point extraction coupled to high performance liquid chromatography for preconcentration and determination of curcumin in human urine.

    Science.gov (United States)

    Rahimi, Marzieh; Hashemi, Payman; Nazari, Fariba

    2014-05-15

    A cold column trapping-cloud point extraction (CCT-CPE) method coupled to high performance liquid chromatography (HPLC) was developed for preconcentration and determination of curcumin in human urine. A nonionic surfactant, Triton X-100, was used as the extraction medium. In the proposed method, a low surfactant concentration of 0.4% v/v and a short heating time of only 2min at 70°C were sufficient for quantitative extraction of the analyte. For the separation of the extraction phase, the resulted cloudy solution was passed through a packed trapping column that was cooled to 0 °C. The temperature of the CCT column was then increased to 25°C and the surfactant rich phase was desorbed with 400μL ethanol to be directly injected into HPLC for the analysis. The effects of different variables such as pH, surfactant concentration, cloud point temperature and time were investigated and optimum conditions were established by a central composite design (response surface) method. A limit of detection of 0.066mgL(-1) curcumin and a linear range of 0.22-100mgL(-1) with a determination coefficient of 0.9998 were obtained for the method. The average recovery and relative standard deviation for six replicated analysis were 101.0% and 2.77%, respectively. The CCT-CPE technique was faster than a conventional CPE method requiring a lower concentration of the surfactant and lower temperatures with no need for the centrifugation. The proposed method was successfully applied to the analysis of curcumin in human urine samples.

  10. Long-term progression and therapeutic response of visceral metastatic disease non-invasively monitored in mouse urine using beta-human choriogonadotropin secreting tumor cell lines.

    Science.gov (United States)

    Francia, Giulio; Emmenegger, Urban; Lee, Christina R; Shaked, Yuval; Folkins, Christopher; Mossoba, Miriam; Medin, Jeffrey A; Man, Shan; Zhu, Zhenping; Witte, Larry; Kerbel, Robert S

    2008-10-01

    Historically, the use of mouse models of metastatic disease to evaluate anticancer therapies has been hampered because of difficulties in detection and quantification of such lesions without sacrificing the mice, which in turn may also be dictated by institutional or ethical guidelines. Advancements in imaging technologies have begun to change this situation. A new method to non-invasively measure tumor burden, as yet untested to monitor spontaneous metastases, is the use of transplanted tumors expressing secretable human beta-chorionic gonadotropin (beta-hCG) that can be measured in urine. We describe examples of beta-hCG-transfected tumor cell lines for evaluating the effect of different therapies on metastatic disease, which in some cases involved monitoring tumor growth for >100 days. We used beta-hCG-tagged mouse B16 melanoma and erbB-2/Her-2-expressing human breast cancer MDA-MB-231 models, and drug treatments included metronomic low-dose cyclophosphamide chemotherapy with or without a vascular endothelial growth factor receptor 2-targeting antibody (DC101) or trastuzumab, the erbB-2/Her-2-targeting antibody. Both experimental and spontaneous metastasis models were studied; in the latter case, an increase in urine beta-hCG always foreshadowed the development of lung, liver, brain, and kidney metastases. Metastatic disease was unresponsive to DC101 or trastuzumab monotherapy treatment, as assessed by beta-hCG levels. Our results also suggest that beta-hCG levels may be set as an end point for metastasis studies, circumventing guidelines, which have often hampered the use of advanced disease models. Collectively, our data indicates that beta-hCG is an effective noninvasive preclinical marker for the long term monitoring of untreated or treated metastatic disease.

  11. Arsenic and other trace elements in groundwater and human urine in Ha Nam province, the Northern Vietnam: contamination characteristics and risk assessment.

    Science.gov (United States)

    Pham, Long Hai; Nguyen, Hue Thi; Van Tran, Cuong; Nguyen, Ha Manh; Nguyen, Tung Hoang; Tu, Minh Binh

    2017-06-01

    The contamination characteristics of arsenic and other trace elements in groundwater and the potential risks of arsenic from the groundwater were investigated. Elevated contamination of arsenic, barium and manganese was observed in tube-well water of two villages (Chuyen Ngoai and Chau Giang) in Ha Nam province in the Northern Vietnam. Concentrations of As in the groundwater ranged from 12.8 to 884 µg/L with mean values in Chuyen Ngoai and Chau Giang were 614.7 and 160.1 µg/L, respectively. About 83 % of these samples contained As concentrations exceeding WHO drinking water guideline of 10 μg/L. The mean values of Mn and Ba in groundwater from Chuyen Ngoai and Chau Giang were 300 and 657 μg/L and 650 and 468 μg/L, respectively. The mean value of Ba concentration in groundwater in both Chuyen Ngoai and Chau Giang was about 22 % of the samples exceeded the WHO guideline (700 µg/L). Arsenic concentrations in human urine of residents from Chuyen Ngoai and Chau Giang were the range from 8.6 to 458 µg/L. The mean values of Mn and Ba in human urine of local people from Chuyen Ngoai were 46.9 and 62.8 μg/L, respectively, while those in people from Chau Giang were 25.9 and 45.9 μg/L, respectively. The average daily dose from ingesting arsenic for consuming both untreated and treated groundwater is from 0.02 to 11.5 and 0.003 to 1.6 μg/kg day, respectively. Approximately, 57 % of the families using treated groundwater and 64 % of the families using untreated groundwater could be affected by elevated arsenic exposure.

  12. Determination of eight illegal drugs in human urine by combination of magnetic solid-phase extraction with capillary zone electrophoresis.

    Science.gov (United States)

    Chen, Ming-Luan; Suo, Li-Li; Gao, Qiang; Feng, Yu-Qi

    2011-08-01

    Using magnetite/silica/poly(methacrylic acid-co-ethylene glycol dimethacrylate) (Fe(3)O(4)/SiO(2)/poly(MAA-co-EDMA)) magnetic microspheres, a rapid and high-throughput magnetic solid-phase extraction coupled with capillary zone electrophoresis (MSPE-CZE) method was developed for the determination of illegal drugs (ketamine, amphetamines, opiates, and metabolites). The MSPE of target analytes could be completed within 2 min, and the eight target analytes could be baseline separated within 15 min by CZE with 30 mM phosphate buffer solution (PBS, pH 2.0) containing 15% v/v ACN as background electrolyte. Furthermore, hydrodynamic injection with field-amplified sample stacking (FASS) was employed to enhance the sensitivity of this MSPE-CZE method. Under such optimal conditions, the limits of detection for the eight target analytes ranged from 0.015 to 0.105 μg/mL. The application feasibility of MSPE-CZE in illegal drugs monitoring was demonstrated by analyzing urine samples, and the recoveries of target drugs for the spiked sample ranging from 85.4 to 110.1%. The method reproducibility was tested by evaluating the intra- and interday precisions, and relative standard deviations of <10.3 and 12.4%, respectively, were obtained. To increase throughput of the analysis, a home-made MSPE array that has potential application to the treatment of 96 samples simultaneously was used.

  13. A rapid, cost-effective method for analyzing organophosphorus pesticide metabolites in human urine for counter-terrorism response.

    Science.gov (United States)

    Weerasekera, Gayanga; Smith, Kimberly D; Needham, Larry L; Barr, Dana B

    2008-01-01

    Organophosphorus (OP) pesticides are used as insecticides in agriculture and pest control and are often called "junior strength" nerve agents because they share the same mechanism of toxicity. OP pesticides are metabolized to dialkylphosphates and other metabolites, which are excreted in urine. In case of a terrorism incident involving widely available OP pesticides, an occurrence that may be likely given their widespread availability, a rapid, accurate, and cost-effective method for detecting exposure is required. We have evaluated several analytical methods to determine the most reliable and cost-effective methods for incident response. Our comparisons have included different internal standards (isotopically labeled standards versus chemically similar surrogate standards), different isolation techniques (some of which are automatable), and different analysis platforms. We found that isotopically labeled standards were a necessity to provide accurate quantification; the chemically similar surrogate was not suitable as an internal standard. The most sensitive and precise method uses isotopically labeled standards with gas chromatography-tandem mass spectrometry analysis. However, the most cost-effective method employed isotopically labeled standards with gas chromatography-single quadrupole-mass spectrometry using a less expensive mass selective detector. Because this method is lower in cost, it may be a more viable option for equipping multiple laboratories with chemical-terrorism response capabilities.

  14. New potential biomarkers for mesterolone misuse in human urine by liquid chromatography quadrupole time-of-flight mass spectrometry.

    Science.gov (United States)

    Lu, Jianghai; Fernández-Álvarez, María; Yang, Sheng; He, Genye; Xu, Youxuan; Aguilera, Rodigo

    2015-01-01

    In this paper, mesterolone metabolic profiles were investigated carefully. Mesterolone was administered to one healthy male volunteer. Urinary extracts were analyzed by liquid chromatography quadruple time-of-flight mass spectrometry (LC-QTOFMS) for the first time. Liquid-liquid extraction was applied to processing urine samples, and dilute-shoot analyses of intact metabolites were also presented. In LC-QTOFMS analysis, chromatographic peaks for potential metabolites were hunt down by using the theoretical [M-H](-) as target ions in full scan experiment, and their actual deprotonated ions were analyzed in targeted MS/MS mode. Ten metabolites including seven new sulfate and three glucuronide conjugates were found for mesterolone. Because of no useful fragment ion for structural elucidation, gas chromatography-mass spectrometry instrumentation was employed to obtain structural details of the trimethylsilylated phase I metabolite released after solvolysis. Thus, their potential structures were proposed particularly by a combined MS approach. All the metabolites were also evaluated in terms of how long they could be detected, and S1 (1α-methyl-5α-androst-3-one-17β-sulfate) together with S2 (1α-methyl-5α-androst-17-one-3β-sulfate) was detected up to 9 days after oral administration, which could be the new potential biomarkers for mesterolone misuse.

  15. Determination of Atropine Sulfate in Human Urines by Capillary Electrophoresis Using Chemical Modified Electrode as Electrochemiluminescence Sensor

    Directory of Open Access Journals (Sweden)

    Min Zhou

    2011-01-01

    Full Text Available A Ru(bpy3 2+-based electrochemiluminescence (ECL detection coupled with capillary electrophoresis (CE was developed for the determination of atropine sulfate on the basis of an Eu-PB modified platinum electrode as the working electrode. The analyte was injected to separation capillary of 50 cm length (25 μm i.d., 360 μm o.d. by electrokinetic injection for 10 s at 10 kV. Parameters related to the separation and detection were discussed and optimized. It was proved that 10 mM phosphate buffer at pH 8.0 could achieve the most favorable resolution, and the high sensitivity of detection was obtained by using the detection potential at 1.15 V and 5 mM Ru(bpy3 2+ in 80 mM phosphate buffer at pH 8.0 in the detection reservoir. Under the optimized conditions, the ECL peak area was in proportion to atropine sulfate concentration in the range from 0.08 to 20 μg⋅mL−1 with a detection limit of 50 ng⋅mL−1 (3σ. The relative standard derivations of migration time and peak area were 0.81 and 3.19%, respectively. The developed method was successfully applied to determine the levels of atropine sulfate in urine samples of patients with recoveries between 90.9 and 98.6%.

  16. Phase I and II metabolites of speciogynine, a diastereomer of the main Kratom alkaloid mitragynine, identified in rat and human urine by liquid chromatography coupled to low- and high-resolution linear ion trap mass spectrometry.

    Science.gov (United States)

    Philipp, Anika A; Wissenbach, Dirk K; Weber, Armin A; Zapp, Josef; Maurer, Hans H

    2010-11-01

    Mitragyna speciosa (Kratom in Thai), a Thai medical plant, is misused as herbal drug of abuse. Besides the most abundant alkaloids mitragynine (MG) and paynantheine (PAY), several other alkaloids were isolated from Kratom leaves, among them the third abundant alkaloid is speciogynine (SG), a diastereomer of MG. The aim of this present study was to identify the phase I and II metabolites of SG in rat urine after the administration of a rather high dose of the pure alkaloid and then to confirm these findings using human urine samples after Kratom use. The applied liquid chromatography coupled to low- and high-resolution mass spectrometry (LC-HRMS-MS) provided detailed information on the structure in the MS(n) mode particularly with high resolution. For the analysis of the human samples, the LC separation had to be improved markedly allowing the separation of SG and its metabolites from its diastereomer MG and its metabolites. In analogy to MG, besides SG, nine phase I and eight phase II metabolites could be identified in rat urine, but only three phase I and five phase II metabolites in human urine. These differences may be caused by the lower SG dose applied by the user of Kratom preparations. SG and its metabolites could be differentiated in the human samples from the diastereomeric MG and its metabolites comparing the different retention times determined after application of the single alkaloids to rats. In addition, some differences in MS(2) and/or MS(3) spectra of the corresponding diastereomers were observed.

  17. Comparative estimation of use potentialities of salt-accumulating and salt-eliminating halophytes for inclusion of NaCl contained in human mineralized urine in BLSS's mass exchange

    Science.gov (United States)

    Tikhomirova, Natalia; Ushakova, Sofya; Kudenko, Yurii; Griboskaya, Illiada; Shklavtsova, Ekaterina; Balnokin, Yurii; Popova, Larissa; Myasoedov, Nikolay; Gros, Jean-Bernard; Lasseur, Christophe

    Comparative potentialities of different halophytes' cultivation on a human mineralized urine containing NaCl with the aim of this salt inclusion into the intrasystem BLSS mass exchange were investigated. Two halophyte species were studied namely, salt-accumulating (Salicornia europaea) and salt-eliminating (Limonium gmelinii). During the first two vegetation weeks the plants had been grown on the Knop solution; then a daily norm of the human mineralized urine was gradually added in the experiment solutions. During vegetation the model solutions simulating the urine mineral composition were gradually added in the control solutions. The NaCl concentration in the experiment and control solutions of the first treatment was 9 g/l and that of the second treatment was 20 g/l. The mineralized human urine exposed some inhibitory action on Salicornia europaea and Limonium gmelinii plants. The experiment plants' productivity was lower in comparison with the control. As far as Limonium gmelinii appears to be a perennial plant the growth rate and productivity of this halophyte species was signifi- cantly lower in comparison with Salicornia europaea. Na content in Salicornia europaea plants was higher in comparison with sodium amount emitted by Limonium gmelinii. Consequently Salicornia europaea appears to be a more perspective halophyte for its further use in BLSS aiming at involvement of sodium chloride contained in human liquid wastes in intrasystem mass exchange.

  18. A simple LC-MS/MS method for the determination of cortisol, cortisone and tetrahydro-metabolites in human urine: assay development, validation and application in depression patients.

    Science.gov (United States)

    Zhai, Xuejia; Chen, Fen; Zhu, Chaoran; Lu, Yongning

    2015-03-25

    Chronic stress as well as major depressive disorders is associated with cortisol metabolism. Two enzymes modulate cortisol (F) and cortisone (E) interconversion: 11β-hydroxysteroid dehydrogenase type 1 and type 2 (11β-HSD1 and 11β-HSD2). Furthermore, F and E were inactivated by 5α and 5β reductases to their tetrahydro-metabolites: tetrahydrocortisol (THF), allo-tetrahydrocortisol (5α-THF) and tetrahydrocortisone (THE). To better understand depression a LC-MS/MS method for simultaneous determination of F, E THF, 5α-THF and THE in human urine has been developed and validated. The quantification range was 0.1-160 ng mL(-1) for F and E, and 0.2-160 ng mL(-1) for the tetrahydro-metabolites, with >86.1% recovery for all analytes. The nocturnal urine concentrations of F, E and tetrahydro-metabolites in 12 apparently healthy male adult volunteers and 12 drug-free male patients (age range, 20-50 years) with a diagnosis of depression were analyzed. A series of significant changes in glucocorticoid metabolism can be detected: F/E ratios and (THF+5α-THF)/THE ratios as well as F and THF concentrations were significantly higher in depression patients than in healthy subjects (p<0.05); 5α-THF/F ratios, 5α-THF/THF ratios as well as 5α-THF concentrations were significantly lower in depression patients (p<0.05). The results pointed to the decreased 11β-HSD2 activity and a dysfunction in the 5α-reductase pathway in depressed patients. This method allows the assessment of 11β-HSD1/2 and 5α/β-reductase activities in a single analytical run providing an innovative tool to explain the potential etiology of depression. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Inter- and intraindividual correlations of background abundances of (2)H, (18)O and (17)O in human urine and implications for DLW measurements.

    Science.gov (United States)

    Berman, E S F; Melanson, E L; Swibas, T; Snaith, S P; Speakman, J R

    2015-10-01

    The method of choice for measuring total energy expenditure in free-living individuals is the doubly labeled water (DLW) method. This experiment examined the behavior of natural background isotope abundance fluctuations within and between individuals over time to assess possible methods of accounting for variations in the background isotope abundances to potentially improve the precision of the DLW measurement. In this work, we measured natural background variations in (2)H, (18)O and (17)O in water from urine samples collected from 40 human subjects who resided in the same geographical area. Each subject provided a urine sample for 30 consecutive days. Isotopic abundances in the samples were measured using Off-Axis Integrated Cavity Output Spectroscopy. Autocorrelation analyses demonstrated that the background isotopes in a given individual were not temporally correlated over the time scales of typical DLW studies. Using samples obtained from different individuals on the same calendar day, cross-correlation analyses demonstrated that the background variations of different individuals were not correlated in time. However, the measured ratios of the three isotopes (2)H, (18)O and (17)O were highly correlated (R(2)=0.89-0.96). Although neither specific timing of DLW water studies nor intraindividual comparisons were found to be avenues for reducing the impact of background isotope abundance fluctuations on DLW studies, strong inter-isotope correlations within an individual confirm that use of a dosing ratio of 8‰:1‰ (0.6 p.p.m.: p.p.m.) optimizes DLW precision. Theoretical implications for the possible use of (17)O measurements within a DLW study require further study.

  20. Study on the determination and chiral inversion of R-salbutamol in human plasma and urine by liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Zhou, Ting; Zeng, Jing; Liu, Shan; Zhao, Ting; Wu, Jie; Lai, Wenshi; He, Mingzhi; Xu, Beining; Qu, Shanshan; Xu, Ling; Tan, Wen

    2015-10-01

    The chiral inversion has been a concerned issue during the research and development of a chiral drug. In this study, a sensitive chiral liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for determination of salbutamol enantiomers in human plasma and urine. The chiral inversion mechanism of R-salbutamol was fully investigated for the first time by studying the effects of physicochemical factors, including pH, temperature and time. A fitted model to predict the chiral inversion ratio of R-salbutamol was proposed using a Box-Behnken design. All the samples were separated on an Astec Chirobiotic T column and detected by a tandem mass spectrometer in multiple reaction monitoring mode. Lower limit of quantification of 0.100ng/mL was achieved under the optimized conditions. The method was fully validated and successfully applied to the clinical pharmacokinetic study of R-salbutamol in healthy volunteers. Chiral inversion of R-salbutamol to S-salbutamol has been detected in urine samples. The results indicated that pH and temperature were two dominant factors that caused the chiral inversion of R-salbutamol, which should be taken into consideration during the analysis of chiral drugs. The chiral inversion of R-salbutamol determined in this study was confirmed resulted from the gastric acid in stomach rather than caused by the analysis conditions. Moreover, the calculated results of the fitted model matched very well with the enantioselective pharmacokinetic study of R-salbutamol, and the individual difference of the chiral inversion ratio of R-salbutamol was related to the individual gastric environment. On the basis of the results, this study provides important and concrete information not only for the chiral analysis but also for the metabolism research of chiral drugs.

  1. HPLC Determination of α-keto Acids in Human Serum and Urine after Derivatization with 4-Nitro-1,2-phenylenediamine

    Directory of Open Access Journals (Sweden)

    Shamroz Bano Sahito

    2013-06-01

    Full Text Available The determination of α-keto acids has clinical importance, because these are intermediates in a number of biochemical processes. This work reports the development of an HPLC procedure for the analysis α-keto acids in blood and urine samples after derivatization with 4-nitro-1,2-phenylenediamine (NPD. Nine α-keto acids: glyoxylic acid (GA, pyruvic acid (PYR, 2-oxobutyric acid (KB, 3-methyl-2-oxobutyric acid (MKBA, 3-methyl-2-oxovaleric acid (K3MVA, 2-oxoglutaric acid (KG, 4-methyl-2-oxovaleric acid (K4MVA, 2-oxohexanoic acid (KHA and phenylpyruvic acid (PPY were derivatized with (NPD at pH 3 and separated on a Zorbax 300 SB-C18 HPLC column (4.6x150mm id and photodiode array detection at 255 nm. The isocratic elution was performed with methanol: water: acetonitrile (42: 56:2, v/ v/ v with a flow rate 0.9 mL/min. The keto acids separated within 14 min. The method was repeatable with a relative standard deviation (RSD of 0.1-2.9% for each of the α-keto acids. The limits of detection and quantitation were obtained within the range 0.05-0.26 µg/ mL and 0.15-0.8 µg/ mL respectively. The method was applied for determination of α-keto acids from a pharmaceutical preparation, human serum and urine samples of healthy volunteers and diabetic patients. The results were further confirmed by standard addition technique. The method is rapid and simple and is suitable for the separation and determination of α-keto acids from clinical samples.

  2. Second-order multivariate models for the processing of standard-addition synchronous fluorescence-pH data. Application to the analysis of salicylic acid and its major metabolite in human urine.

    Science.gov (United States)

    Pagani, Ariana P; Ibañez, Gabriela A

    2014-05-01

    In the present work, we describe the determination of salicylic acid and its major metabolite, salicyluric acid, in spiked human urine samples, using synchronous fluorescence spectra measured in a flow-injection system with a double pH gradient. Because the fluorescent urine background constitutes a potentially interfering signal, it becomes necessary to achieve the second-order advantage. Moreover, due to significant changes in the signal of the analytes in the presence of the urine matrix, mainly for salicyluric acid, standard addition was required in order to obtain appropriate quantifications. Several second-order multivariate calibration models were evaluated for this purpose: PARAFAC and MCR-ALS in two different modes, and PLS/RBL.

  3. Simple and sensitive high-performance liquid chromatographic method for the determination of 1,5-benzodiazepine clobazam and its active metabolite N-desmethylclobazam in human serum and urine with application to 1,4-benzodiazepines analysis.

    Science.gov (United States)

    Kunicki, P K

    2001-01-05

    A HPLC-UV determination of clobazam and N-desmethylclobazam in human serum and urine is presented. After simple liquid-liquid extraction with dichloromethane the compounds and an internal standard diazepam were separated on a Supelcosil LC-8-DB column at ambient temperature under isocratic conditions using the mobile phase: CH3CN-water-0.5 M KH2PO4-H3PO4 (440:540:20:0.4, v/v and 360:580:60:0.4, v/v for serum and urine, respectively). The detection was performed at 228 nm with limits of quantification of 2 ng/ml for serum and 1 ng/ml for urine. Relative standard deviations for intra- and inter-assay precision were found below 8% for both compounds for all the tested concentrations. The described procedure may be easily adapted for several 1,4-benzodiazepines.

  4. Vortex-assisted liquid-liquid extraction combined with field-amplified sample injection and sweeping micellar electrokinetic chromatography for improved determination of β-blockers in human urine.

    Science.gov (United States)

    Jouyban, Abolghasem; Sorouraddin, Mohammad Hossein; Farajzadeh, Mir Ali; Somi, Mohammad Hossein; Fazeli-Bakhtiyari, Rana

    2016-01-01

    A new micellar electrokinetic chromatography (MEKC) method was developed and validated for the analysis of carvedilol and propranolol in human urine samples. In this study, vortex-assisted liquid-liquid extraction (VALLE) coupled with field-amplified sample injection and sweeping was employed for biological sample clean-up and sensitivity enhancement in MEKC. After VALLE, the urine samples were analyzed by MEKC. Tris-phosphate buffer (60mmolL(-1), pH 2.0) containing 40% (v/v) methanol was first filled into an uncoated fused-silica capillary (56cm, 50µm i.d.). The pretreated urine sample was loaded by electrokinetic injection (10kV, 250s). The stacking and separation were performed using Tris-phosphate buffer (30mmolL(-1), pH 3.0) containing 30% (v/v) methanol and 50mmolL(-1) sodium dodecyl sulfate (SDS) at -25kV. Detection was carried out at 195 and 214nm for carvedilol and propranolol, respectively. The suggested method is linear (r(2)≥0.997) over a dynamic range of 0.005-1µgmL(-1) in urine. The intra- and inter-day relative standard deviation and relative error values of the method were below 20%, which shows good precision and accuracy. Finally, this method was successfully applied to the analysis of real urine samples.

  5. HPLC法测定人尿液中奥拉西坦的浓度%Determination of Oxiracetam in Human Urine by HPLC

    Institute of Scientific and Technical Information of China (English)

    刘秀菊; 张志清; 孙倩; 亢泽坤

    2013-01-01

    OBJECTIVE: To establish the method for the determination of oxiracetam in human urine, and to provide reference for the study of metabolic pathways of oxiracetam. METHODS: After precipitated with methanol, HPLC method was adopted for the content determination. With acyclovir as an internal standard, a Symmetry shied? RP18 column was used with mobile phase consisted of acetonitrile-water (5:95) at the flow rate of 0.5 ml/min. The column temperature was 40 ℃ and the detection wavelength was set at 210 nm. The injection volume was 20 μl. RESULTS: The linear ranges of oxiracetam were 20-2 400 mg/L. Absolute recoveries of urine at low, middle and high concentrations were (90.38 ± 4.93)% , (93.71 ± 3.54)% and (95.04 ± 2.31)% , and the method recoveries were (98.32 ± 1.96)% , (104.99 ± 1.74)% and (103.4 ± 2.43)%. The intra-day RSDs of urine at three concentrations were 3.40% , 4.76% and 1.04% , and inter-day RSDs were 2.82% , 3.64% and 1.90% , respectively. Trial data showed that 32.89% original drug discharged in 24 h after oral administration of Oxiracetam capsules (1600 mg). CONCLUSION: The method is simple, accurate, sensitive, and can be used for the pharmacokinetic study of oxiracetam in urine.%目的:建立测定人尿液中奥拉西坦浓度的方法,为奥拉西坦代谢途径的研究提供参考.方法:尿液以甲醇沉淀蛋白后,以高效液相色谱法进样测定,其中色谱柱为Symmetry shiedTMRP18柱,流动相为乙腈-水(5∶95),流速为0.5 ml/min,检测波长为210nm,柱温为40℃,进样量为20μl,内标为阿昔洛韦.结果:奥拉西坦尿药浓度在20~2400mg/L范围内线性关系良好;低、中、高3种浓度尿液样品的绝对回收率分别为(90.38±4.93)%、(93.71±3.54)%、(95.04±2.31)%,方法回收率分别为(98.32±1.96)%、(104.99±1.74)%、(103.4±2.43)%;日内RSD分别为3.40%、4.76%、1.04%,日间RSD分别为2.82%、3.64%、1.90%;试验数据显示人口服奥拉西坦胶囊(1600mg

  6. 气相色谱法快速分析人血、尿中地芬尼多%Determination of difenidol in human blood and urine by GC

    Institute of Scientific and Technical Information of China (English)

    刘缙; 黄飞; 高丽伟; 蔡向阳; 金鸣

    2011-01-01

    目的 建立人血、尿中地芬尼多的气相色谱快速分析方法.方法用氯仿提取血、尿中的地芬尼多,采用气相色谱法进行定性、定量分析.以正常人血浆、尿液为空白样本,分别添加标准地芬尼多及SKF525(内标物)对方法进行考察和优化,并对1例大剂量误服地芬尼多中毒者体液浓度进行快速测定和检测.结果所建方法分析血、尿中地芬尼多的线性范围均为5.0~200.0μg/mL;最低检测限均为1.0μg/mL(S/N≥3); 日内、日间精密度RSD≤5.6%(n=5);回收率:血≤(106.23±2.05)%;尿≤(104.19±5.51)%.结论 该分析方法操作便捷、实用,适用于地芬尼多的临床血药浓度快速监测和法医毒物鉴定.%Objective The thesis aims at developing the method for the quick analysis of gas chromatography(GC) of difenidol in human blood and urine.Methods Difenidol was extracted from blood or urine samples with chloroform and analyzed by the GC method qualitatively and quantitatively.The standard difenidol and SKF525 ( internal standard) were added to blank samples of blood and urine with the aim of testing and optimizing the method.The GC method was successfully applied to quickly detect and test the body fluid concentration of the victim who had taken difenidol hydrochloride by mistake.Results The linear range were 5.0 ~ 200.0μg/mL with R2 = 0.996 5 in blood and R2 = 0.997 2 in urine.The limit of detection was 1.0μg/mL(S/N ≥3).The intra- and inter- day precision of assay for difenidol were less than 5.6%.The recoveries of difenidol were less than ( 106.23 ± 2.05 ) % in blood and ( 104.19 ± 5.51 ) %.Conclusion The GC method was proved to be easy and helpful in the aspects of the quick detection of difenidol concentration in blood and the forensic toxicological analysis.

  7. Rapid and sensitive analysis of phthalate metabolites, bisphenol A, and endogenous steroid hormones in human urine by mixed-mode solid-phase extraction, dansylation, and ultra-performance liquid chromatography coupled with triple quadrupole mass spectrometry.

    Science.gov (United States)

    Wang, He-xing; Wang, Bin; Zhou, Ying; Jiang, Qing-wu

    2013-05-01

    Steroid hormone levels in human urine are convenient and sensitive indicators for the impact of phthalates and/or bisphenol A (BPA) exposure on the human steroid hormone endocrine system. In this study, a rapid and sensitive method for determination of 14 phthalate metabolites, BPA, and ten endogenous steroid hormones in urine was developed and validated on the basis of ultra-performance liquid chromatography coupled with electrospray ionization triple quadrupole mass spectrometry. The optimized mixed-mode solid phase-extraction separated the weakly acidic or neutral BPA and steroid hormones from acidic phthalate metabolites in urine: the former were determined in positive ion mode with a methanol/water mobile phase containing 10 mM ammonium formate; the latter were determined in negative ion mode with a acetonitrile/water mobile phase containing 0.1 % acetic acid, which significantly alleviated matrix effects for the analysis of BPA and steroid hormones. Dansylation of estrogens and BPA realized simultaneous and sensitive analysis of the endogenous steroid hormones and BPA in a single chromatographic run. The limits of detection were less than 0.84 ng/mL for phthalate metabolites and less than 0.22 ng/mL for endogenous steroid hormones and BPA. This proposed method had satisfactory precision and accuracy, and was successfully applied to the analyses of human urine samples. This method could be valuable when investigating the associations among endocrine-disrupting chemicals, endogenous steroid hormones, and relevant adverse outcomes in epidemiological studies.

  8. Studies on the metabolism of mitragynine, the main alkaloid of the herbal drug Kratom, in rat and human urine using liquid chromatography-linear ion trap mass spectrometry.

    Science.gov (United States)

    Philipp, Anika A; Wissenbach, Dirk K; Zoerntlein, Siegfried W; Klein, Oliver N; Kanogsunthornrat, Jidapha; Maurer, Hans H

    2009-08-01

    Mitragynine (MG) is an indole alkaloid of the Thai medicinal plant Mitragyna speciosa (Kratom in Thai) and reported to have opioid agonistic properties. Because of its stimulant and euphoric effects, Kratom is used as a herbal drug of abuse. The aim of the presented study is to identify the phase I and II metabolites of MG in rat and human urine after solid-phase extraction (SPE) using liquid chromatography-linear ion trap mass spectrometry providing detailed structure information in the MSn mode particularly with high resolution. The seven identified phase I metabolites indicated that MG was metabolized by hydrolysis of the methylester in position 16, O-demethylation of the 9-methoxy group and of the 17-methoxy group, followed, via the intermediate aldehydes, by oxidation to carboxylic acids or reduction to alcohols and combinations of some steps. In rats, four metabolites were additionally conjugated to glucuronides and one to sulfate, but in humans, three metabolites to glucuronides and three to sulfates.

  9. Flow injection on-line dilution for multi-element determination in human urine with detection by inductively coupled plasma mass spectrometry

    DEFF Research Database (Denmark)

    Wang, Jianhua; Hansen, Elo Harald; Gammelgaard, Bente

    2001-01-01

    of 16.5 with on-line standard addition, and sup/ 103/Rh was used as internal standard to compensate for signal fluctuation. The procedure was validated by the analysis of two standard reference materials SRM 2670 (NIST) and Seronorm(TM) Trace Elements in Urine. Recovery experiments were performed...... urine samples. No correlations between the concentrations of the elements were observed....

  10. Prevalence and antifungal susceptibility of Cryptococcus neoformans isolated from pigeon excreta in Chon Buri Province, Eastern Thailand.

    Science.gov (United States)

    Tangwattanachuleeporn, Marut; Somparn, Poorichaya; Poolpol, Kulwara; Gross, Uwe; Weig, Michael; Bader, Oliver

    2013-01-01

     The prevalence of cerebral meningitis caused by Cryptococcus neoformans in HIV-infected patients in Eastern Thailand is high. However, little is known about the occurrence of this pathogenic yeast in the environment of this region.  The aim of our study was to characterize the prevalence of C. neoformans, its serotypes and antifungal drug susceptibilities in environmental isolates from Chon Buri, Eastern Thailand.  C. neoformans was isolated from 10% of fifty pigeon excreta examined from this province. All C. neoformans isolates were of serotype A and although the isolates displayed slightly decreased susceptibility towards fluconazole, all tested sensitive to amphotericin B, fluconazole and itraconazole. This study is the first report of the occurrence of C. neoformans in pigeon excreta in eastern Thailand.

  11. Prevalence of human papillomavirus infection in the oropharynx and urine among sexually active men: a comparative study of infection by papillomavirus and other organisms, including Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma spp., and Ureaplasma spp.

    Science.gov (United States)

    Nakashima, Kazufumi; Shigehara, Kazuyoshi; Kawaguchi, Shohei; Wakatsuki, Akira; Kobori, Yoshitomo; Nakashima, Kazuyoshi; Ishii, Yasunori; Shimamura, Masayoshi; Sasagawa, Toshiyuki; Kitagawa, Yasuhide; Mizokami, Atsushi; Namiki, Mikio

    2014-01-27

    Oropharyngeal squamous cell carcinoma (OSCC) has shown a gradual increase in male predominance due to the increasing incidence of human papillomavirus (HPV)-associated OSCC. However, the mode of HPV transmission to the oral cavity is poorly understood, and little is known about the epidemiology of oral HPV infection in men. The prevalence rates of HPV, Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma spp., and Ureaplasma spp. were compared in the oropharynx (oral cavity) and urine of male Japanese patients attending a sexually transmitted disease clinic. The study population consisted of 213 men aged 16 - 70 years old (mean: 34.4 years old). Oropharyngeal gargles and urine were collected, and sedimented cells were preserved in liquid-based cytology solution. After DNA extraction, β-globin and infectious organisms were analyzed by a PCR-based method. The HPV genotype was determined by HPV GenoArray test. β-Globin was positive in 100% and 97.7% of oral and urine samples, respectively. HPV detection rates were 18.8% and 22.1% in oral and urine samples, respectively, suggesting that the prevalence of HPV infection in the oral cavity was similar to that in the urinary tract. N. gonorrhoeae was more prevalent in oral (15.6%) than urine samples (9.1%), whereas C. trachomatis was detected more frequently in urine (15.9%) than oral samples (4.2%). The detection rates of M. genitalium, M. hominis, and Ureaplasma spp. were 5.2%, 10.3%, and 16.0% in oral samples, and 7.7%, 6.3%, and 19.2% in urine, respectively. There were no significant differences in the detection rates of Mycoplasma spp. and Ureaplasma spp. between anatomical locations. The distribution of HPV types were similar in oral and urine samples, and HPV16 was the most common type. The majority of men with HPV infection in both the oral cavity and urine had concordant oral and urinary HPV infection. The presence of urinary HPV infection was an independent risk factor of oral HPV infection, with an odds

  12. Biogenic and toxic elements in feathers, eggs, and excreta of Gentoo penguin (Pygoscelis papua ellsworthii) in the Antarctic.

    Science.gov (United States)

    Metcheva, Roumiana; Yurukova, Lilyana; Teodorova, Svetla E

    2011-11-01

    Feathers, eggs, and excreta of Gentoo penguin (Pygoscelis papua ellsworthii), adults, from Livingston Island (South Shetlands), chosen as bioindicators, were used to test the quality of the Antarctic environment. Sex was not examined. The bioaccumulations of toxic trace elements (Cd, Pb, Al, and As), essential trace elements (Fe, Cu, Zn, Mn, Cr, V, Ni, and Sr), and major essential elements (Na, K, Mg, Ca, P, and S) were established. For the first time data about the element contents in Gentoo eggs is provided. Two hypotheses were tested: (1) there are differences in the metal levels among eggs and feathers; and (2) the element concentrations are highest in the excreta. The hypotheses were confirmed at 0.01-0.05 confidence levels. The concentrations of almost all trace elements were significantly higher in the feathers compared to those in the eggs. The following values of the concentrations ratio Fe/Zn were obtained: in the embryo, Fe/Zn = 1.5, and in the feathers, Fe/Zn = 0.5. The concentration of Pb in the embryo and excreta was below 0.4 μg/g, and Cd and As in eggs were below 0.05 and 0.3 μg/g, respectively. This indicates that there is no toxic risk for penguin offspring. Arsenic could be considered as a potential pollutant for Antarctic soil due to its relative high concentration in excreta, 5.13 μg/g. The present data (year 2007) were compared to the data for years 2002 and 2003. No trend of toxic element contamination was established. The concentrations of Pb, Cd, and As in representatives from the top of the food chain in the Antarctic (the present study) and Arctic (literature data) were compared. The data supports the hypothesis that there is an abnormality in cadmium levels in polar marine areas. Regarding Pb, the South Shetlands displayed 3-fold lower level compared to the Aleutians.

  13. Effects of dietary supplementation with Gynura procumbens (Merr.) on egg yolk cholesterol, excreta microflora and laying hen performance.

    Science.gov (United States)

    Lokhande, A; Ingale, S L; Lee, S H; Sen, S; Khong, C; Chae, B J; Kwon, I K

    2014-01-01

    Abstract 1. The present study investigated the effects of dietary supplementation with Gynura procumbens on egg yolk and serum cholesterol and triglycerides, excreta microflora, laying performance and egg quality. 2. A total of 160 Hy-Line Brown layers (45 weeks old) were randomly assigned into 4 treatments on the basis of laying performance. Each treatment had 4 replicates with 10 birds each. 3. Dietary treatments were basal diet supplemented with 0 (control), 2.5, 5.0 and 7.5 g/kg diet G. procumbens during 56-d feeding period. 4. Serum (d 21, 42 and 56) and egg yolk (d 28, 42 and 56) cholesterol and triglycerides concentrations were linearly reduced with increasing dietary concentrations of G. procumbens. 5. Increasing dietary concentrations of G. procumbens linearly reduced the excreta total anaerobic bacteria (d 28), Clostridium sp. and Escherichia coli (d 28 and 56) populations. 6. Overall egg production and egg mass were linearly increased, and overall feed efficiency was linearly improved with increase in dietary G. procumbens. 7. Dietary increasing concentrations of G. procumbens linearly improved egg yolk colour (d 28 and 56) and breaking strength of eggs (d 56). 8. The results obtained in the present experiment indicate that dietary supplementation with G. procumbens could reduce the egg yolk cholesterol, suppresses harmful excreta microflora and improves layers performance.

  14. First metabolic profile of PV8, a novel synthetic cathinone, in human hepatocytes and urine by high-resolution mass spectrometry.

    Science.gov (United States)

    Swortwood, Madeleine J; Ellefsen, Kayla N; Wohlfarth, Ariane; Diao, Xingxing; Concheiro-Guisan, Marta; Kronstrand, Robert; Huestis, Marilyn A

    2016-07-01

    Novel psychoactive substances (NPS) are ever changing on the drug market, making it difficult for toxicology laboratory methods to stay current with so many new drugs. Recently, PV8, a synthetic pyrrolidinophenone, was detected in seized products in Japan (2013), The Netherlands (2014), and Germany (2014). There are no controlled PV8 administration studies, and no pharmacodynamic and pharmacokinetic data. The objective was to determine PV8's metabolic stability in human liver microsome (HLM) incubation and its metabolism following human hepatocyte incubation and high-resolution mass spectrometry (HRMS) with a Thermo Scientific Q-Exactive. Data were acquired with a full-scan data-dependent mass spectrometry method. Scans were thoroughly data mined with different data processing algorithms and analyzed in WebMetaBase. PV8 exhibited a relatively short 28.8 min half-life, with an intrinsic 24.2 μL/min/mg microsomal clearance. This compound is predicted to be an intermediate clearance drug with an estimated human 22.7 mL/min/kg hepatic clearance. Metabolic pathways identified in vitro included: hydroxylation, ketone reduction, carboxylation, N-dealkylation, iminium formation, dehydrogenation, N-oxidation, and carbonylation. The top three in vitro metabolic pathways were di-hydroxylation > ketone reduction > γ-lactam formation. Authentic urine specimen analyses revealed the top three metabolic pathways were aliphatic hydroxylation > ketone reduction + aliphatic hydroxylation > aliphatic carboxylation, although the most prominent peak was parent PV8. These data provide useful urinary metabolite targets (aliphatic hydroxylation, aliphatic hydroxylation + ketone reduction, aliphatic carboxylation, and di-hydroxylation) for forensic and clinical testing, and focus reference standard companies' synthetic efforts to provide commercially available standards needed for PV8 biological specimen testing. Graphical Abstract Top four PV8 metabolites identified in vitro

  15. Simultaneous determination of parabens, alkylphenols, phenylphenols, bisphenol A and triclosan in human urine, blood and breast milk by continuous solid-phase extraction and gas chromatography-mass spectrometry.

    Science.gov (United States)

    Azzouz, Abdelmonaim; Rascón, Andrés J; Ballesteros, Evaristo

    2016-02-05

    A highly sensitive gas chromatography-mass spectrometry (GC-MS) method for the determination of endocrine disrupting chemicals (EDCs) including parabens, alkylphenols, phenylphenols, bisphenol A and triclosan in human breast milk, blood and urine samples is proposed. Blood and milk require a pretreatment to remove proteins and other substances potentially interfering with the continuous solid-phase extraction (SPE) system used; on the other hand, urine samples can be directly introduced into the system after filtering. Analytes are retained on a LiChrolut EN column and derivatized by silylation following elution with acetonitrile. The resulting trimethylsilyl derivatives are determined by GC-MS. The proposed method exhibited good linearity (r(2)>0.995) for all target EDCs over the concentration range 0.7-10,000ng/l in urine, and 3.3-50,000ng/l in blood and milk. Also, it provided low limits of detection (0.2-1.8ng/l in urine, and 1.0-9.0ng/l in blood and milk), good precision (relative standard deviations less than 7%) and recoveries from 86 to 104%. A total of 24 human fluid samples were analyzed and most found to contain some target EDC at concentrations from 0.10 to 14μg/l.

  16. FUNGI ISOLATED FROM THE EXCRETA OF WILD BIRDS IN SCREENING CENTERS IN PELOTAS, RS, BRAZIL

    Directory of Open Access Journals (Sweden)

    Josiara Furtado Mendes

    2014-12-01

    Full Text Available The identification of the fungal species belonging to the healthy microflora in animals is a precondition for the recognition of pathological processes causing them. The aim of this study was to investigate the presence of potentially pathogenic fungi in the feces of wild birds collected in Screening Centers. Samples were collected from the feces of 50 cages with different species of birds. The samples were processed according to the modified method STAIB and the plates incubated at 32 °C for up to ten days with daily observation for detection of fungal growth. The isolation of the following species was observed: Malassezia pachydermatis, Candida albicans, C. famata, C. guilliermondii, C. sphaerica, C. globosa, C. catenulata, C. ciferri, C. intermedia, Cryptococcus laurentii, Trichosporon asahii, Geotrichum klebahnii, Aspergillus spp., A. niger and Penicillium spp. Knowing the character of some opportunistic fungi is important in identifying them, facilitating the adoption of preventive measures, such as proper cleaning of cages, since the accumulation of excreta may indicate a risk for both health professionals and centers for screening public health.

  17. Container-based sanitation: assessing costs and effectiveness of excreta management in Cap Haitien, Haiti.

    Science.gov (United States)

    Tilmans, Sebastien; Russel, Kory; Sklar, Rachel; Page, Leah; Kramer, Sasha; Davis, Jennifer

    2015-04-01

    Container-based sanitation (CBS) - in which wastes are captured in sealable containers that are then transported to treatment facilities - is an alternative sanitation option in urban areas where on-site sanitation and sewerage are infeasible. This paper presents the results of a pilot household CBS service in Cap Haitien, Haiti. We quantify the excreta generated weekly in a dense urban slum,((1)) the proportion safely removed via container-based public and household toilets, and the costs associated with these systems. The CBS service yielded an approximately 3.5-fold decrease in the unmanaged share of faeces produced, and nearly eliminated the reported use of open defecation and "flying toilets" among service recipients. The costs of this pilot small-scale service were higher than those of large-scale waterborne sewerage, but economies of scale have the potential to reduce CBS costs over time. The paper concludes with a discussion of planning and policy implications of incorporating CBS into the menu of sanitation options for rapidly growing cities.

  18. Survival of Ascaris eggs and hygienic quality of human excreta in Vietnamese composting latrines

    DEFF Research Database (Denmark)

    Jensen, Peter K. M.; Phuc, Pham D.; Konradsen, Flemming;

    2009-01-01

    this is the length of time that farmers have available to produce fertilizer between two cropping seasons. This study aimed to investigate whether hygienically safe fertilizer could be produced in the latrines within this period of time. Methods: By inoculating eggs of the helminth parasite indicator Ascaris suum......, could therefore potentially provide a hygienic acceptable fertilizer....

  19. Practice of using human excreta as fertilizer and implications for health in Nghean Province, Vietnam

    DEFF Research Database (Denmark)

    Phuc, P. D.; Konradsen, Flemming; Phuong, P. T.

    2006-01-01

    using latrine wastes as fertilizers in a community in central Vietnam. Information was collected through structured questionnaire interviews administered to 75 farming households, focus group discussions, and key informant interviews. The majority (64%) of households had a single vault latrine......, a possession that was associated with low income (chi2= 12.45; p lime likely to increase pH and pathogen die-off. About 28...

  20. A gas chromatography-mass spectrometry method for the quantitation of N-nitrosoproline and N-acetyl-S-allylcysteine in human urine: application to a study of the effects of garlic consumption on nitrosation.

    Science.gov (United States)

    Cope, Keary; Seifried, Harold; Seifried, Rebecca; Milner, John; Kris-Etherton, Penny; Harrison, Earl H

    2009-11-15

    Biomarkers in urine can provide useful information about the bioactivation of chemical carcinogens and can be used to investigate the chemoprotective properties of dietary nutrients. N-Nitrosoproline (NPRO) excretion has been used as an index for endogenous nitrosation. In vitro and animal studies have reported that compounds in garlic may suppress nitrosation and inhibit carcinogenesis. We present a new method for extraction and sensitive detection of both NPRO and N-acetyl-S-allylcysteine from urine. The latter is a metabolite of S-allylcysteine, which is found in garlic. Urine was acidified and the organic acids were extracted by reversed-phase extraction (RP-SPE) and use of a polymeric weak anion exchange (WAX-SPE) resin. NPRO was quantified by isotope dilution gas chromatography-mass spectrometry (GC-MS) using [13C5]NPRO and N-nitrosopipecolinic acid (NPIC) as internal standards. This method was used to analyze urine samples from a study that was designed to test whether garlic supplementation inhibits NPRO synthesis. Using this method, 2.4 to 46.0 ng NPRO/ml urine was detected. The method is straightforward and reliable, and it can be performed with readily available GC-MS instruments. N-Acetyl-S-allylcysteine was quantified in the same fraction and detectable at levels of 4.1 to 176.4 ng/ml urine. The results suggest that 3 to 5 g of garlic supplements inhibited NPRO synthesis to an extent similar to a 0.5-g dose of ascorbic acid or a commercial supplement of aged garlic extract. Urinary NPRO concentration was inversely associated with the N-acetyl-S-allylcysteine concentration. It is possible that allyl sulfur compounds found in garlic may inhibit nitrosation in humans.