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Sample records for high-performance thin-layer chromatography

  1. High performance thin layer chromatography profile of Cassytha filiformis

    Institute of Scientific and Technical Information of China (English)

    Mythili Sathiavelu; Sathiavelu Arunachalam

    2012-01-01

    Objective: To study the phenols, flavonoids, saponin profile of the medicinal plant Cassytha filiformis (C. filiformis) using high performance thin layer chromatography (HPTLC). Methods:The extracts were tested to determine the presence of various phytochmeicals like alkaloids, phenolic compounds, flavonoids, carbohydrates, glycosides, saponins, terpenoids, tannins, fixed oils, fats and protein and aminoacids (Harborne and Harborne, 1998). HPTLC studies were carried out by Harborne and Wagner et al method. Different compositions of the mobile phase for HPTLC analysis were tested in order to obtain high resolution and reproducible peaks. Results: The results of the preliminary phytochemical studies confirm the presence of phenols, alkaloids, carbohydrates, saponins, flavanoids, terpenoids and tannins in the methanolic extracts of C. filiformis. The methanolic extracts of C. filiformis displayed the presence of 13 types of phenolic substances with 13 different Rf values ranging from 0.01 to 0.96. The results illustrated the presence of 9 different types of flavonoides with 9 different Rf values ranging from 0.01 to 0.97. The results of HPTLC analysis of saponins demonstrated the presence of 11 different types of saponins with 11 different Rf values ranging from 0.04 to 0.92. Conclusions: In the present study we observed the phenols, flavonoids, saponin profile of the medicinal plant C. filiformis using high performance thin layer chromatography (HPTLC). Hence it was concluded that the phenolic compounds present in the methonolic extract could be responsible for antioxidant activities. Plant derived antioxidants, especially phenols and flavonoids, have been described to have various properties like anticancer, antiaging and prevention of cardiovascular diseases. Furthur, separation and characterization of the bioactive compound from the plant is to be evaluated and reported in near future.

  2. Phytochemical analysis of ethanolic extract of Dichrostachys Cinerea W and Arn leaves by a thin layer chromatography, high performance thin layer chromatography and column chromatography

    OpenAIRE

    M Vijayalakshmi; K Periyanayagam; K Kavitha; K Akilandeshwari

    2013-01-01

    Background: The leaves of Dichrostachys cinerea are used as laxative, diuretic, painkiller. It is also used in the treatment of gonorrhoea, boils, oedema, gout, veneral diseases and nasopharyngeal affections, etc. Materials and Methods: The Phytochemical investigation of ethanolic extract of D. cinerea leaves were performed by standard chemical tests, thin layer chromatography (TLC) by using various solvent systems, and by high performance liquid chromatography (HPTLC). Two compounds were...

  3. Pressurized planar electrochromatography, high-performance thin-layer chromatography and high-performance liquid chromatography--comparison of performance.

    Science.gov (United States)

    Płocharz, Paweł; Klimek-Turek, Anna; Dzido, Tadeusz H

    2010-07-16

    Kinetic performance, measured by plate height, of High-Performance Thin-Layer Chromatography (HPTLC), High-Performance Liquid Chromatography (HPLC) and Pressurized Planar Electrochromatography (PPEC) was compared for the systems with adsorbent of the HPTLC RP18W plate from Merck as the stationary phase and the mobile phase composed of acetonitrile and buffer solution. The HPLC column was packed with the adsorbent, which was scrapped from the chromatographic plate mentioned. An additional HPLC column was also packed with adsorbent of 5 microm particle diameter, C18 type silica based (LiChrosorb RP-18 from Merck). The dependence of plate height of both HPLC and PPEC separating systems on flow velocity of the mobile phase and on migration distance of the mobile phase in TLC system was presented applying test solute (prednisolone succinate). The highest performance, amongst systems investigated, was obtained for the PPEC system. The separation efficiency of the systems investigated in the paper was additionally confirmed by the separation of test component mixture composed of six hormones. 2010 Elsevier B.V. All rights reserved.

  4. A validated high performance thin layer chromatography method for determination of yohimbine hydrochloride in pharmaceutical preparations.

    Science.gov (United States)

    Badr, Jihan M

    2013-01-01

    Yohimbine is an indole alkaloid used as a promising therapy for erectile dysfunction. A number of methods were reported for the analysis of yohimbine in the bark or in pharmaceutical preparations. In the present work, a simple and sensitive high performance thin layer chromatographic method is developed for determination of yohimbine (occurring as yohimbine hydrochloride) in pharmaceutical preparations and validated according to International Conference of Harmonization (ICH) guidelines. The method employed thin layer chromatography aluminum sheets precoated with silica gel as the stationary phase and the mobile phase consisted of chloroform:methanol:ammonia (97:3:0.2), which gave compact bands of yohimbine hydrochloride. Linear regression data for the calibration curves of standard yohimbine hydrochloride showed a good linear relationship over a concentration range of 80-1000 ng/spot with respect to the area and correlation coefficient (R(2)) was 0.9965. The method was evaluated regarding accuracy, precision, selectivity, and robustness. Limits of detection and quantitation were recorded as 5 and 40 ng/spot, respectively. The proposed method efficiently separated yohimbine hydrochloride from other components even in complex mixture containing powdered plants. The amount of yohimbine hydrochloride ranged from 2.3 to 5.2 mg/tablet or capsule in preparations containing the pure alkaloid, while it varied from zero (0) to 1.5-1.8 mg/capsule in dietary supplements containing powdered yohimbe bark. We concluded that this method employing high performance thin layer chromatography (HPTLC) in quantitative determination of yohimbine hydrochloride in pharmaceutical preparations is efficient, simple, accurate, and validated.

  5. High performance thin layer chromatography fingerprint analysis of guava (Psidium guajava) leaves

    Science.gov (United States)

    Astuti, M.; Darusman, L. K.; Rafi, M.

    2017-05-01

    High-performance thin layer chromatography (HPTLC) fingerprint analysis is commonly used for quality control of medicinal plants in term of identification and authentication. In this study, we have been developed HPTLC fingerprint analysis for identification of guava (Psidium guajava) leaves raw material. A mixture of chloroform, acetone, and formic acid in the ratio 10:2:1 was used as the optimum mobile phase in HPTLC silica plate and with 13 bands were detected. As reference marker we chose gallic acid (Rf = 0.21) and catechin (Rf = 0.11). The two compound were detected as pale black bands at 366 nm after derivatization with sulfuric acid 10% v/v (in methanol) reagent. Validation of the method was met within validation criteria, so the developed method could be used for quality control of guava leaves.

  6. In situ identification of high-performance thin-layer chromatography spots by fourier transform surface-enhanced Raman scattering

    Science.gov (United States)

    Koglin, Eckhardt; Kramer, Hella; Sawatski, Juergen; Lehner, Carolin; Hellman, Janice L.

    1994-01-01

    FT-SERS has been used to identify samples supported on high-performance thin-layer chromatography plates. The TLC plates were sprayed with colloidal silver solutions which resulted in enhancement of the FT-Raman scattering of these biologically and environmentally important compounds.

  7. HPTLC-aptastaining - Innovative protein detection system for high-performance thin-layer chromatography

    Science.gov (United States)

    Morschheuser, Lena; Wessels, Hauke; Pille, Christina; Fischer, Judith; Hünniger, Tim; Fischer, Markus; Paschke-Kratzin, Angelika; Rohn, Sascha

    2016-05-01

    Protein analysis using high-performance thin-layer chromatography (HPTLC) is not commonly used but can complement traditional electrophoretic and mass spectrometric approaches in a unique way. Due to various detection protocols and possibilities for hyphenation, HPTLC protein analysis is a promising alternative for e.g., investigating posttranslational modifications. This study exemplarily focused on the investigation of lysozyme, an enzyme which is occurring in eggs and technologically added to foods and beverages such as wine. The detection of lysozyme is mandatory, as it might trigger allergenic reactions in sensitive individuals. To underline the advantages of HPTLC in protein analysis, the development of innovative, highly specific staining protocols leads to improved sensitivity for protein detection on HPTLC plates in comparison to universal protein derivatization reagents. This study aimed at developing a detection methodology for HPTLC separated proteins using aptamers. Due to their affinity and specificity towards a wide range of targets, an aptamer based staining procedure on HPTLC (HPTLC-aptastaining) will enable manifold analytical possibilities. Besides the proof of its applicability for the very first time, (i) aptamer-based staining of proteins is applicable on different stationary phase materials and (ii) furthermore, it can be used as an approach for a semi-quantitative estimation of protein concentrations.

  8. A validated high performance thin layer chromatography method for determination of yohimbine hydrochloride in pharmaceutical preparations

    OpenAIRE

    Jihan M Badr

    2013-01-01

    Background: Yohimbine is an indole alkaloid used as a promising therapy for erectile dysfunction. A number of methods were reported for the analysis of yohimbine in the bark or in pharmaceutical preparations. Materials and Method: In the present work, a simple and sensitive high performance thin layer chromatographic method is developed for determination of yohimbine (occurring as yohimbine hydrochloride) in pharmaceutical preparations and validated according to International Conference of Ha...

  9. Quantification of Quercetin and Rutin from Benincasa hispida Seeds and Carissa Congesta Roots by High-performance Thin Layer Chromatography and High-performance Liquid Chromatography.

    Science.gov (United States)

    Doshi, Gaurav Mahesh; Une, Hemant Devidas

    2016-01-01

    In Indian Ayurvedic system, Benincasa hispida (BH) and Carissa congesta (CC) are well-known plants used for major and minor ailments. BH has been regarded as Kushmanda, whereas CC has been used in immune-related disorders of the human system. Quercetin and rutin identified from the vast plethora of plant extracts have proved to possess ethnopharmacological relevance. In present studies, we have determined quercetin and rutin in terms of percentage in BH seeds and CC roots by high-performance thin layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC). After extraction and phytochemical screening, the extracts were subjected to quantification for the presence of quercetin and rutin by HPTLC and HPLC. HPTLC showed quercetin as 44.60, 27.13% and rutin as 32.00, 36.31% w/w, whereas HPLC revealed quercetin as 34.00, 35.00% and rutin as 21.99, 45.03% w/v in BH and CC extracts, respectively. The BH and CC extracts have elucidated peaks that were corresponding with standard peaks on undertaking chromatographic studies. Quercetin and rutin are isolated from BH seeds and CC roots by High Performance. Thin Layer Chromatography and High Performance Liquid Chromatography. HPTLC revealed presence of quercetin as 44.60, 27.13 % and rutin as 32.00, 36.31 % w/w. HPLC revealed presence of quercetin as 34.00, 35.00 % and rutin as 21.99, 45.03 % w/v. Abbreviation Used: HPTLC: High Performance Thin Layer Chromatography; HPLC: High Pressure Liquid Chromatography, UV: Ultraviolet, CC: Carissa congesta, BH: Benincasa hispida.

  10. Quantification of Quercetin and Rutin from Benincasa hispida Seeds and Carissa Congesta Roots by High-performance Thin Layer Chromatography and High-performance Liquid Chromatography

    Science.gov (United States)

    Doshi, Gaurav Mahesh; Une, Hemant Devidas

    2016-01-01

    Objective: In Indian Ayurvedic system, Benincasa hispida (BH) and Carissa congesta (CC) are well-known plants used for major and minor ailments. BH has been regarded as Kushmanda, whereas CC has been used in immune-related disorders of the human system. Quercetin and rutin identified from the vast plethora of plant extracts have proved to possess ethnopharmacological relevance. Materials and Methods: In present studies, we have determined quercetin and rutin in terms of percentage in BH seeds and CC roots by high-performance thin layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC). After extraction and phytochemical screening, the extracts were subjected to quantification for the presence of quercetin and rutin by HPTLC and HPLC. Results: HPTLC showed quercetin as 44.60, 27.13% and rutin as 32.00, 36.31% w/w, whereas HPLC revealed quercetin as 34.00, 35.00% and rutin as 21.99, 45.03% w/v in BH and CC extracts, respectively. Conclusion: The BH and CC extracts have elucidated peaks that were corresponding with standard peaks on undertaking chromatographic studies. SUMMARY Quercetin and rutin are isolated from BH seeds and CC roots by High Performance. Thin Layer Chromatography and High Performance Liquid Chromatography. HPTLC revealed presence of quercetin as 44.60, 27.13 % and rutin as 32.00, 36.31 % w/w. HPLC revealed presence of quercetin as 34.00, 35.00 % and rutin as 21.99, 45.03 % w/v. Abbreviation Used: HPTLC: High Performance Thin Layer Chromatography; HPLC: High Pressure Liquid Chromatography, UV: Ultraviolet, CC: Carissa congesta, BH: Benincasa hispida PMID:26941534

  11. High Performance Thin Layer Chromatography method for analysis of 3,4-methylenedioxymethamphetamine in seized tablets

    Directory of Open Access Journals (Sweden)

    Boris E. Duffau

    2015-12-01

    Full Text Available Context: Consumption of synthetic drugs had increased in recent years, used as a recreational drug by young people who presume that consumption of this drug is harmless for health; however clinical studies have shown that this stimulant and its metabolites are toxic. Due to these reasons, chemical analysis of this illicit drug is crucial from the points of view of occupational medicine, toxicology, and law enforcement with the aim of pursuit the traffic of illegal drug. Aims: Implement and fully validate a rapid and simple method for detection and quantitation of MDMA by High-Performance Thin Layer Chromatography in seized samples. Methods: With the implemented method was analyzed 12 positive samples seized by Chilean police, to found the concentration of MDMA in ecstasy tablets. Results: The method was fully validated, the linearity of the method was evaluated by the calibration curve between 51.0 – 510.0 µg/band (R2 0.9977; limit of detection was 12.1 µg per band, and limit of quantitation was 36.8 µg per band. The precision of the method (RSD was lower than 5.0%. Accuracy was evaluated by determination of the percentage of MDMA recovered by the assay (99.13%, and relative Uncertainty was 6.66%. With this method, it was analyzed real seized samples of MDMA, results showed that all samples contained MDMA and concentration was between 18.15 – 59.84 % w/w. Conclusions: The method is selective, sensitive, and specific, with possible application in forensic analysis. To the best of our knowledge, this is the first report about concentration of MDMA in ecstasy pills in Chile.

  12. Separation of pigment formulations by high-performance thin-layer chromatography with automated multiple development.

    Science.gov (United States)

    Stiefel, Constanze; Dietzel, Sylvia; Endress, Marc; Morlock, Gertrud E

    2016-09-02

    Food packaging is designed to provide sufficient protection for the respective filling, legally binding information for the consumers like nutritional facts or filling information, and an attractive appearance to promote the sale. For quality and safety of the package, a regular quality control of the used printing materials is necessary to get consistently good print results, to avoid migration of undesired ink components into the food and to identify potentially faulty ink batches. Analytical approaches, however, have hardly been considered for quality assurance so far due to the lack of robust, suitable methods for the analysis of rarely soluble pigment formulations. Thus, a simple and generic high-performance thin-layer chromatography (HPTLC) method for the separation of different colored pigment formulations was developed on HPTLC plates silica gel 60 by automated multiple development. The gradient system provided a sharp resolution for differently soluble pigment constituents like additives and coating materials. The results of multi-detection allowed a first assignment of the differently detectable bands to particular chemical substance classes (e.g., lipophilic components), enabled the comparison of different commercially available pigment batches and revealed substantial variations in the composition of the batches. Hyphenation of HPTLC with high resolution mass spectrometry and infrared spectroscopy allowed the characterization of single unknown pigment constituents, which may partly be responsible for known quality problems during printing. The newly developed, precise and selective HPTLC method can be used as part of routine quality control for both, incoming pigment batches and monitoring of internal pigment production processes, to secure a consistent pigment composition resulting in consistent ink quality, a faultless print image and safe products. Hyphenation of HPTLC with the A. fischeri bioassay gave first information on the bioactivity or rather

  13. Combining surface enhanced Raman scattering (SERS) and high-performance thin-layer chromatography (HPTLC)

    Science.gov (United States)

    Koglin, E.

    A new method for preparing SERS active surfaces using silver colloidal spheres deposited on HPTLC plates, used for thin-layer chromatography, is discussed in detail. The sensitivity of these activated HPTLC plates is so high that in-situ vibrational investigations of chromatogram spots are possible at the nanogram level. The HPTLC/SERS spectra of purine, benzoic acid and 1-nitro-pyrene adsorbed on silver colloidal activated silica gel plates are measured in the nanogram region. In addition we also report in this paper on the results of a feasibility study performed to evaluate the analytical potential of micro-Raman spectroscopy (triple monochromator, multichannel detection system) in SERS/HPTLC spot characterization. It permits the acquisition of Raman spectra from HPTLC spots down to 1 μm in size or other forms of microsamples approaching the picogram level in mass.

  14. Effect-directed analysis via hyphenated high-performance thin-layer chromatography for bioanalytical profiling of sunflower leaves.

    Science.gov (United States)

    Móricz, Ágnes M; Ott, Péter G; Yüce, Imanuel; Darcsi, András; Béni, Szabolcs; Morlock, Gertrud E

    2018-01-19

    High-performance thin-layer chromatography (HPTLC) coupled with effect-directed analysis was used for non-targeted screening of sunflower leaf extract for components exhibiting antioxidant, antibacterial and/or cholinesterase enzyme inhibitory effects. The active compounds were characterized by HPTLC-electrospray ionization-high resolution mass spectrometry (ESI-HRMS) and HPTLC-Direct Analysis in Real Time (DART)-MS/MS. The latter ambient ionization technique (less soft than ESI) resulted in oxidation and fragmentation products and characteristic fragment ions. NMR spectroscopy after targeted isolation via preparative normal phase flash chromatography and semi-preparative reversed phase high-performance liquid chromatography supported the identification of two diterpenes to be (-)-kaur-16-en-19-oic acid and 15-α-angeloyloxy-ent-kaur-16-en-19-oic acid. Both compounds found to be multi-potent as they inhibited acetylcholinesterase and butyrylcholinesterase and showed antibacterial effects against Gram-positive Bacillus subtilis and Gram-negative Aliivibrio fischeri bacteria. Kaurenoic acid was also active against the Gram-negative pepper pathogenic Xanthomonas euvesicatoria bacteria. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Development and validation of high-performance liquid chromatography and high-performance thin-layer chromatography methods for the quantification of khellin in Ammi visnaga seed

    Science.gov (United States)

    Kamal, Abid; Khan, Washim; Ahmad, Sayeed; Ahmad, F. J.; Saleem, Kishwar

    2015-01-01

    Objective: The present study was used to design simple, accurate and sensitive reversed phase-high-performance liquid chromatography RP-HPLC and high-performance thin-layer chromatography (HPTLC) methods for the development of quantification of khellin present in the seeds of Ammi visnaga. Materials and Methods: RP-HPLC analysis was performed on a C18 column with methanol: Water (75: 25, v/v) as a mobile phase. The HPTLC method involved densitometric evaluation of khellin after resolving it on silica gel plate using ethyl acetate: Toluene: Formic acid (5.5:4.0:0.5, v/v/v) as a mobile phase. Results: The developed HPLC and HPTLC methods were validated for precision (interday, intraday and intersystem), robustness and accuracy, limit of detection and limit of quantification. The relationship between the concentration of standard solutions and the peak response was linear in both HPLC and HPTLC methods with the concentration range of 10–80 μg/mL in HPLC and 25–1,000 ng/spot in HPTLC for khellin. The % relative standard deviation values for method precision was found to be 0.63–1.97%, 0.62–2.05% in HPLC and HPTLC for khellin respectively. Accuracy of the method was checked by recovery studies conducted at three different concentration levels and the average percentage recovery was found to be 100.53% in HPLC and 100.08% in HPTLC for khellin. Conclusions: The developed HPLC and HPTLC methods for the quantification of khellin were found simple, precise, specific, sensitive and accurate which can be used for routine analysis and quality control of A. visnaga and several formulations containing it as an ingredient. PMID:26681890

  16. Method development for Lawsone estimation in Trichup herbal hair powder by high-performance thin layer chromatography

    Directory of Open Access Journals (Sweden)

    Maunang M Patel

    2013-01-01

    Full Text Available A simple, specific, accurate, precise and robust high-performance thin-layer chromatographic method has been developed and validated for estimation of Lawsone in Trichup herbal hair powder (coded as a THHP, polyherbal formulation. The chromatographic development was carried out on aluminum plates pre-coated with silica gel 60F 254 and good resolution was achieved with Toluene: Ethyl acetate: Glacial acetic acid (8:1:1 v/v/v as mobile phase. Lawsone detection was carried out densitometrically at 277 nm and obtained retardation factor value was 0.46 ± 0.02. The method was validated with respect to specificity, linearity, accuracy, precision and robustness. The calibration curve was achieved to be linear over a range of 5-60 μg/ml and regression coefficient was obtained 0.998. Accuracy of chromatographic method was evaluated by standard addition method; recovery was obtained 99.25 ± 0.61% . The peak purity of Lawsone was achieved 0.999 r. Relative standard deviation for intraday and inter-day precision was 0.37-0.56% and 0.42-0.55%, respectively. The limit of detection and limit of quantification of the Lawsone were found to be 1.08 μg/m land 3.28 μg/ml, respectively. This result shows that the method was well validated. In the present study, the Lawsone content was found 0.322 ± 0.014% in THHP. This study reveals that the proposed high performance thin layer chromatography method is accurate, fast and cost- effective for routine estimation of Lawsone in polyherbal formulation.

  17. Assessment of aflatoxin B1 in livestock feed and feed ingredients by high-performance thin layer chromatography

    Directory of Open Access Journals (Sweden)

    Korrapati Kotinagu

    2015-12-01

    Full Text Available Aim: Detection of aflatoxin B1 in Livestock compound Feed and feed ingredients by high-performance thin layer chromatography (HPTLC. Materials and Methods: Chromatography was performed on HPTLC silica gel 60 F 254, aluminum sheets by CAMAG automatic TLC sampler 4, with mobile phase condition chloroform:acetone:water (28:4:0.06. Extraction of aflatoxin B1 from samples was done as per AOAC method and screening and quantification done by HPTLC Scanner 4 under wavelength 366 nm. Results: A total of 97 livestock feed (48 and feed ingredients (49 samples received from different livestock farms and farmers were analyzed for aflatoxin B1of which 29 samples were contaminated, constituting 30%. Out of 48 livestock compound feed samples, aflatoxin B1 could be detected in 16 samples representing 33%, whereas in livestock feed ingredients out of 49 samples, 13 found positive for aflatoxin B1 representing 24.5%. Conclusion: HPTLC assures good recovery, precision, and linearity in the quantitative determination of aflatoxin B1 extracted from Livestock compound feed and feed ingredients. As more number of feed and feed ingredients are contaminated with aflatoxin B1 which causes deleterious effects in both animal and human beings, so there is a need for identifying the source of contamination, executing control measures, enabling better risk assessment techniques, and providing economic benefits.

  18. Characterization of E 471 food emulsifiers by high-performance thin-layer chromatography-fluorescence detection.

    Science.gov (United States)

    Oellig, Claudia; Brändle, Klara; Schwack, Wolfgang

    2018-07-13

    Mono- and diacylglycerol (MAG and DAG) emulsifiers, also known as food additive E 471, are widely used to adjust techno-functional properties in various foods. Besides MAGs and DAGs, E 471 emulsifiers additionally comprise different amounts of triacylglycerols (TAGs) and free fatty acids (FFAs). MAGs, DAGs, TAGs and FFAs are generally determined by high-performance liquid chromatography (HPLC) or gas chromatography (GC) coupled to mass selective detection, analyzing the individual representatives of the lipid classes. In this work we present a rapid and sensitive method for the determination of MAGs, DAGs, TAGs and FFAs in E 471 emulsifiers by high-performance thin-layer chromatography with fluorescence detection (HPTLC-FLD), including a response factor system for quantitation. Samples were simply dissolved and diluted with t-butyl methyl ether before a two-fold development was performed on primuline pre-impregnated LiChrospher silica gel plates with diethyl ether and n-pentane/n-hexane/diethyl ether (52:20:28, v/v/v) as the mobile phases to 18 and 75 mm, respectively. For quantitation, the plate was scanned in the fluorescence mode at UV 366/>400 nm, when the cumulative signal for each lipid class was used. Calibration was done with 1,2-distearin and amounts of lipid classes were calculated with response factors and expressed as monostearin, distearin, tristearin and stearic acid. Limits of detection and quantitation were 1 and 4 ng/zone, respectively, for 1,2-distearin. Thus, the HPTLC-FLD approach represents a simple, rapid and convenient screening alternative to HPLC and GC analysis of the individual compounds. Visual detection additionally enables an easy characterization and the direct comparison of emulsifiers through the lipid class pattern, when utilized as a fingerprint. Copyright © 2018 Elsevier B.V. All rights reserved.

  19. Forensic Analysis of Blue Ball point Pen Inks on Questioned Documents by High Performance Thin Layer Chromatography Technique (HPTLC)

    International Nuclear Information System (INIS)

    Lee, L.C.; Siti Mariam Nunurung; Abdul Aziz Ishak

    2014-01-01

    Nowadays, crimes related to forged documents are increasing. Any erasure, addition or modification in the document content always involves the use of writing instrument such as ball point pens. Hence, there is an evident need to develop a fast and accurate ink analysis protocol to solve this problem. This study is aimed to determine the discrimination power of high performance thin layer chromatography (HPTLC) technique for analyzing a set of blue ball point pen inks. Ink samples deposited on paper were extracted using methanol and separated via a solvent mixture of ethyl acetate, methanol and distilled water (70: 35: 30, v/ v/ v). In this method, the discrimination power of 89.40 % was achieved, which confirm that the proposed method was able to differentiate a significant number of pen-pair samples. In addition, composition of blue pen inks was found to be homogeneous (RSD < 2.5 %) and the proposed method showed good repeatability and reproducibility (RSD < 3. 0%). As a conclusion, HPTLC is an effective tool to separate blue ball point pen inks. (author)

  20. A novel high-performance thin layer chromatography method for quantification of long chain aliphatic hydrocarbons from Cissus quadrangularis

    Directory of Open Access Journals (Sweden)

    Vandana Jain

    2016-08-01

    Full Text Available Context: A high-performance thin layer chromatography (HPTLC is an analytical technique, which can be used for the determination of constituents or marker components in various parts of the plants. Earlier studies have estimated phytoconstituents from the stem and other aerial plant parts of Cissus quadrangularis Linn. Estimation of hydrocarbons can also be successfully done using HPTLC technique using suitable derivatization. Aims: To develop and validate a simple and rapid method for the estimation of long chain aliphatic hydrocarbons from the leaves of C. quadrangularis using HPTLC technique. Methods: Precoated silica gel 60 F254 plates were used as stationary phase. The mobile phase used was hexane (100 %. The detection of spots was carried out using berberine sulphate as detecting reagent. Results: The method was validated in terms of linearity, sensitivity, accuracy, and precision. Linearity range was found to be 2-10 µg/mL, limit of detection 0.127 µg/mL, and limit of quantification 0.384 µg/mL. Conclusions: A novel, simple, accurate, precise and sensitive HPTLC method has been developed and validated for the estimation of long chain aliphatic hydrocarbons obtained from the leaves of C. quadrangularis Linn.

  1. A High-Performance Thin Layer Chromatography (HPTLC) Method for Simultaneous Determination of Diphenhydramine Hydrochloride and Naproxen Sodium in Tablets.

    Science.gov (United States)

    Bhole, R P; Shinde, S S; Chitlange, S S; Wankhede, S B

    2015-01-01

    A rapid and simple high-performance thin layer chromatography (HPTLC) method with densitometry at 230 nm was developed and validated for simultaneous determination of diphenhydramine hydrochloride (DPH) and naproxen sodium (NPS) from pharmaceutical preparation. The separation was carried out on aluminum plates precoated with silica gel 60 F254 using mobile phase toluene:methanol:glacial acetic acid (7.5:1:0.2, v/v/v). The linearity range lies between 200 and 1200 ng/band for DPH and 1760 and 10,560 ng/band for NPS with correlation coefficients of 0.994 and 0.995, respectively. The R f value for DPH is 0.20 ± 0.05 and for NPS is 0.61 ± 0.06. % Recoveries of DPH and NPS was in the range of 99.70%-99.95% and 99.63%-99.95%, respectively. Limit of detection value for DPH was 13.21 ng/band and for NPS was 8.03 ng/band. Limit of quantitation value for DPH was 40.06 ng/band and for NPS was 24.34 ng/band. The developed method was validated as per ICH guidelines. In stability testing, DPH was found unstable to acid and alkaline hydrolysis, and DPH and NPS were found unstable to oxidation, whereas both the drugs were stable to neutral and photodegradation. The proposed method was successfully applied for the routine quantitative analysis of dosage form containing DPH and NPS.

  2. Qualitative and quantitative two-dimensional thin-layer chromatography/high performance liquid chromatography/diode-array/electrospray-ionization-time-of-flight mass spectrometry of cholinesterase inhibitors.

    Science.gov (United States)

    Mroczek, Tomasz

    2016-09-10

    Recently launched thin-layer chromatography-mass spectrometry (TLC-MS) interface enabling extraction of compounds directly from TLC plates into MS ion source was unusually extended into two-dimensional thin-layer chromatography/high performance liquid chromatography (2D, TLC/HPLC) system by its a direct connection to a rapid resolution 50×2.1mm, I.D. C18 column compartment followed by detection by diode array (DAD) and electrospray ionisation time-of-flight mass spectrometry (ESI-TOF-MS). In this way, even not separated bands of complicated mixtures of natural compounds could be analysed structurally, only within 1-2min after development of TLC plates. In comparison to typically applied TLC-MS interface, no ion suppression for acidic mobile phases was observed. Also, substantial increase in ESI-TOF-MS sensitivities and quality of spectra, were noticed. It has been utilised in combination with TLC- based bioautographic approaches of acetylcholinesterase (AChE) inhibitors, However, it can be also applied in any other procedures related to bioactivity (e.g. 2,2-Diphenyl-1-picryl-hydrazyl-DPPH screen test for radicals). This system has been also used for determination of half maximal inhibitory concentration (IC50 values) of the active inhibitor-galanthamine, as an example. Moreover, AChE inhibitory potencies of some of purified plant extracts, never studied before, have been quantitatively measured. This is first report of usage such the 2D TLC/HPLC/MS system both for qualitative and quantitative evaluation of cholinesterase inhibitors in biological matrices. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Marker based standardization of polyherbal formulation (SJT-DI-02 by high performance thin layer chromatography method

    Directory of Open Access Journals (Sweden)

    Bhakti J Ladva

    2014-01-01

    purity of the bands due to marker compounds in the sample extracts were confirmed by overlaying the absorption spectra recorded at start, middle and end position of the band in the sample tracks. After conforming all these things fingerprints were developed for all three formulations which will be act as authentification and quality control tool. Results: % w/w of asarones is 3.61, % w/w of marmelosin is 4.60, % w/w of gallic acid is 10.80 and % w/w of lupeol is 4.13.The method was validated in terms of linearity, precision, repeatability, limit of detection, limit of quantification and accuracy. In well-developed mobile phase system linearity was found to be in the range of 0.983-0.995, % recovery was found to be in the range of 97.48-99.63, % RSD for intraday and interday was found to be 0.13- 0.70 and 0.32 -1.41 and LOD and LOQ was found to be in the range of 0.15- 0.61 and 0.45 -1.83 microgram per ml. Conclusion: Thus High performance thin layer chromatography (HPTLC methods were developed and validated in terms of linearity, precision, repeatability, limit of detection, limit of quantification and accuracy. The methods were rapid, sensitive, reproducible and economical. It does not suffer any positive or negative interference due to common other component present in the formulation and would also serve as a tool for authentication of herbal products containing marmelosin, gallic acid, lupeol and asarones. Thus this work provides standardized and therapeutically active polyherbal formulations for the different ailments.

  4. Quantitative Determination of L-DOPA in Seeds of Mucuna Pruriens Germplasm by High Performance Thin Layer Chromatography.

    Science.gov (United States)

    Raina, Archana P; Khatri, Renu

    2011-07-01

    Mucuna pruriens Linn. is an important medicinal plant used for treatment of Parkinson's disease and many others in ancient Indian medical system. L-DOPA extracted from seeds of Mucuna is a constituent of more than 200 indigenous drug formulations and is more effective as drug than the synthetic counterpart. A densitometric high performance thin-layer chromatographic (HPTLC) method was developed for quantification of L-DOPA content present in the seeds extract. The method involves separation of L-DOPA on precoated silica gel 60 GF(254) HPTLC plates using a solvent system of n-butanol-acetic-acid-water (4:1:1, v/v) as the mobile phase. Quantification was done at 280 nm using absorbance reflectance mode. Linearity was found in the concentration range of 100 to 1000 ng/spot with the correlation coefficient value of 0.9980. The method was validated for accuracy, precision and repeatability. Mean recovery was 100.89%. The LOD and LOQ for L-DOPA determination were found to be 3.41 ng/spot and 10.35 ng/spot respectively. The proposed HPTLC method was found to be precise, specific and accurate for quantitative determination of L-DOPA. It can be used for rapid screening of large germplasm collections of Mucuna pruriens for L-DOPA content. The method was used to study variation in fifteen accessions of Mucuna germplasm collected from different geographical regions.

  5. Thin-layer chromatography coupled with high performance liquid chromatography for determining tetrabromobisphenol A/S and their derivatives in soils.

    Science.gov (United States)

    Liu, Aifeng; Shen, Zhaoshuang; Tian, Yong; Shi, Rongguang; Liu, Yi; Zhao, Zongshan; Xian, Mo

    2017-12-01

    As brominated flame retardants (BFRs), tetrabromobisphenol A/S (TBBPA/S) and their derivatives have raised wide concerns owing to their widely usage, distributions and adverse effects on human health, thus monitoring these BFRs was urgently needed. In this study, a rapid and cost-effective method based on thin-layer chromatography (TLC) sample pre-treatment coupled with high performance liquid chromatography-diode array detector (HPLC-DAD) (UV=214nm) was developed for determining TBBPA/S and their derivatives in soils, including TBBPA, TBBPA bis(allyl ether) (TBBPA-BAE), TBBPA bis(2,3-dibromopropyl ether) (TBBPA-BDBPE), TBBPS bis(allyl ether) (TBBPS-BAE) and TBBPS bis(2,3-dibromopropyl ether) (TBBPS-BDBPE). The method detection limits (MDLs) and the method quantification limits (MQLs) for these BFRs ranged from 0.023 to 0.087μgg -1 dw and 0.076-0.29μgg -1 dw, respectively. The recoveries were 41-108% and both RSD of repeatability and intermediate precision were less than 11%. The developed method presented good performance for analyzing natural soil samples collected from BFRs industrial park, suggesting its great application potential for monitoring environmental TBBPA/S and their derivatives. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Therapeutic drug monitoring of nevirapine in saliva in Uganda using high performance liquid chromatography and a low cost thin-layer chromatography technique

    NARCIS (Netherlands)

    Lamorde, M.; Fillekes, Q.; Sigaloff, K.; Kityo, C.; Buzibye, A.; Kayiwa, J.; Merry, C.; Nakatudde-Katumba, L.; Burger, D.M.; Wit, T.F. de

    2014-01-01

    BACKGROUND: In resource limited settings access to laboratory monitoring of HIV treatment is limited and therapeutic drug monitoring is generally unavailable. This study aimed to evaluate nevirapine concentrations in saliva using low-cost thin-layer chromatography (TLC) and nevirapine concentrations

  7. Therapeutic drug monitoring of nevirapine in saliva in Uganda using high performance liquid chromatography and a low cost thin-layer chromatography technique

    NARCIS (Netherlands)

    Lamorde, Mohammed; Fillekes, Quirine; Sigaloff, Kim; Kityo, Cissy; Buzibye, Allan; Kayiwa, Joshua; Merry, Concepta; Nakatudde-Katumba, Lillian; Burger, David; Rinke de Wit, Tobias F.

    2014-01-01

    In resource limited settings access to laboratory monitoring of HIV treatment is limited and therapeutic drug monitoring is generally unavailable. This study aimed to evaluate nevirapine concentrations in saliva using low-cost thin-layer chromatography (TLC) and nevirapine concentrations in plasma

  8. Two-dimensional high-performance thin-layer chromatography of tryptic bovine albumin digest using normal- and reverse-phase systems with silanized silica stationary phase.

    Science.gov (United States)

    Gwarda, Radosław Łukasz; Dzido, Tadeusz Henryk

    2013-10-18

    Among many advantages of planar techniques, two-dimensional (2D) separation seems to be the most important for analysis of complex samples. Here we present quick, simple and efficient two-dimensional high-performance thin-layer chromatography (2D HPTLC) of bovine albumin digest using commercial HPTLC RP-18W plates (silica based stationary phase with chemically bonded octadecyl ligands of coverage density 0.5μmol/m(2) from Merck, Darmstadt). We show, that at low or high concentration of water in the mobile phase comprised methanol and some additives the chromatographic systems with the plates mentioned demonstrate normal- or reversed-phase liquid chromatography properties, respectively, for separation of peptides obtained. These two systems show quite different separation selectivity and their combination into 2D HPTLC process provides excellent separation of peptides of the bovine albumin digest. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. Quantitative determination of seven chemical constituents and chemo-type differentiation of chamomiles using high-performance thin-layer chromatography.

    Science.gov (United States)

    Sagi, Satyanarayanaraju; Avula, Bharathi; Wang, Yan-Hong; Zhao, Jianping; Khan, Ikhlas A

    2014-10-01

    A simple and rapid high-performance thin-layer chromatographic method was developed for the separation and determination of six flavonoids (rutin, luteolin-7-O-β-glucoside, chamaemeloside, apigenin-7-O-β-glucoside, luteolin, apigenin) and one coumarin, umbelliferone from chamomile plant samples and dietary supplements. The separation was achieved on amino silica stationary phase using dichloromethane/acetonitrile/ethyl formate/glacial acetic acid/formic acid (11:2.5:3:1.25:1.25 v/v/v/v/v) as the mobile phase. The quantitation of each compound was carried out using densitometric reflection/absorption mode at their respective absorbance maxima after postchromatographic derivatization using natural products reagent (1% w/v methanolic solution of diphenylboric acid-β-ethylamino ester). The method was validated for specificity, limits of detection and quantification, precision (intra- and interday) and accuracy. The limits of detection and quantification were found to be in the range from 6-18 and 16-55 ng/band for six flavonoids and one coumarin, respectively. The intra- and interday precision was found to be 90%. The data acquired from high-performance thin-layer chromatography was processed by principal component analysis using XLSTAT statistical software. Application of principal component analysis and agglomerative hierarchial clustering was successfully able to differentiate two chamomiles (German and Roman) and Chrysanthemum. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Determination of Four Major Saponins in Skin and Endosperm of Seeds of Horse Chestnut (Aesculus Hippocastanum L.) Using High Performance Liquid Chromatography with Positive Confirmation by Thin Layer Chromatography

    OpenAIRE

    Abudayeh, Zead Helmi Mahmoud; Al Azzam, Khaldun Mohammad; Naddaf, Ahmad; Karpiuk, Uliana Vladimirovna; Kislichenko, Viktoria Sergeevna

    2015-01-01

    urpose: To separate and quantify four major saponins in the extracts of the skin and the endosperm of seeds of horse chestnut (Aesculus hippocastanum L.) using ultrasonic solvent extraction followed by a high performance liquid chromatography-diode array detector (HPLC-DAD) with positive confirmation by thin layer chromatography (TLC). Methods: The saponins: escin Ia, escin Ib, isoescin Ia and isoescin Ib were extracted using ultrasonic extraction method. The optimized ex...

  11. Development and validation of High-performance Thin-layer Chromatography Method for Simultaneous Determination of Polyphenolic Compounds in Medicinal Plants.

    Science.gov (United States)

    Jayachandran Nair, C V; Ahamad, Sayeed; Khan, Washim; Anjum, Varisha; Mathur, Rajani

    2017-12-01

    Quantitative standardization of plant-based products is challenging albeit essential to maintain their quality. This study aims to develop and validate high-performance thin-layer chromatography (HPTLC) method for the simultaneous determination of rutin (Ru), quercetin (Qu), and gallic acid (Ga) from Psidium guajava Linn. (PG) and Aegle marmelos (L.) Correa. (AM) and correlate with antioxidant activity. The stock solution (1 mg/mL) of standard Ru, Qu, and Ga in methanol: Water (1:1) was serially diluted and spotted (5 μL) on slica gel 60 F 254 thin-layer chromatography plates. Toluene: Ethyl acetate: Formic acid: Methanol (3:4:0.8:0.7, v/v/v) was selected as mobile phase for analysis at 254 nm. Hydroalcoholic (1:1) extracts of leaves of PG and AM were fractionated and similarly analyzed. Antioxidant activity was also determined using 2, 2-diphenyl-1-picrylhydrazyl assay. The developed method was robust and resolved Ru, Qu, and Ga at R f 0.08 ± 0.02, 0.76 ± 0.01, and 0.63 ± 0.02, respectively. The intra-day, interday precision, and interanalyst were limit of detection and limit of quantification for Ru, Qu, and Ga were 4.51, 4.2, 5.27, and 13.67, 12.73, 15.98 ng/spot, respectively. Antioxidant activity (Log 50% inhibition) of PG and AM was 4.947 ± 0.322 and 6.498 ± 0.295, respectively. The developed HPTLC method was rapid, accurate, precise, reproducible, and specific for the simultaneous estimation of Ru, Qu, and Ga. HPTLC method for simultaneous determination and quantification of Rutin, Quercetin and Gallic acid, is reported for quality control of herbal drugs. Abbreviations Used: A: Aqueous fraction; AM: Aegle marmelos L. Correa; B: Butanol fraction; C: Chloroform fraction; EA: Ethyl acetate fraction; Ga: Gallic acid; H: Hexane fraction; HA: Hydroalcoholic extract; HPTLC: High-performance thin-layer chromatography; PG: Psidium guajava ; Qu: Quercetin; Ru: Rutin.

  12. A simple and rapid method for the determination of taxol produced by fungal endophytes from medicinal plants using high performance thin layer chromatography.

    Science.gov (United States)

    Gangadevi, V; Muthumary, J

    2008-01-01

    Taxol is an important anticancer drug used widely in the clinical field. In this study, some endophytic fungi were isolated from selected medicinal plants, and were screened for their potential in the production of taxol, using a rapid separation technique of high performance thin layer chromatography (HPTLC). Of the 20 screened fungi, only 13 fungal species produced taxol in the artificial culture medium. The results of HPTLC showed that the 13 fungal species had identical ultraviolet (UV) characteristics, positive reactivity with a spray reagent, yielding a blue spot, which turned to dark gray after 24 hours, and had Rf values identical to that of the authentic taxol. The amount of taxol was also quantified by comparing the peak area and the peak height of the fungal samples with those of authentic taxol.

  13. Separation, identification and quantification of carotenoids and chlorophylls in dietary supplements containing Chlorella vulgaris and Spirulina platensis using High Performance Thin Layer Chromatography.

    Science.gov (United States)

    Hynstova, Veronika; Sterbova, Dagmar; Klejdus, Borivoj; Hedbavny, Josef; Huska, Dalibor; Adam, Vojtech

    2018-01-30

    In this study, 14 commercial products (dietary supplements) containing alga Chlorella vulgaris and cyanobacteria Spirulina platensis, originated from China and Japan, were analysed. UV-vis spectrophotometric method was applied for rapid determination of chlorophylls, carotenoids and pheophytins; as degradation products of chlorophylls. High Performance Thin-Layer Chromatography (HPTLC) was used for effective separation of these compounds, and also Atomic Absorption Spectrometry for determination of heavy metals as indicator of environmental pollution. Based on the results obtained from UV-vis spectrophotometric determination of photosynthetic pigments (chlorophylls and carotenoids), it was confirmed that Chlorella vulgaris contains more of all these pigments compared to the cyanobacteria Spirulina platensis. The fastest mobility compound identified in Chlorella vulgaris and Spirulina platensis using HPTLC method was β-carotene. Spectral analysis and standard calibration curve method were used for identification and quantification of separated substances on Thin-Layer Chromatographic plate. Quantification of copper (Cu 2+ , at 324.7 nm) and zinc (Zn 2+ , at 213.9nm) was performed using Flame Atomic Absorption Spectrometry with air-acetylene flame atomization. Quantification of cadmium (Cd 2+ , at 228.8 nm), nickel (Ni 2+ , at 232.0nm) and lead (Pb 2+ , at 283.3nm) by Electrothermal Graphite Furnace Atomic Absorption Spectrometry; and quantification of mercury (Hg 2+ , at 254nm) by Cold Vapour Atomic Absorption Spectrometry. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Therapeutic drug monitoring of nevirapine in saliva in Uganda using high performance liquid chromatography and a low cost thin-layer chromatography technique.

    Science.gov (United States)

    Lamorde, Mohammed; Fillekes, Quirine; Sigaloff, Kim; Kityo, Cissy; Buzibye, Allan; Kayiwa, Joshua; Merry, Concepta; Nakatudde-Katumba, Lillian; Burger, David; de Wit, Tobias F Rinke

    2014-09-01

    In resource limited settings access to laboratory monitoring of HIV treatment is limited and therapeutic drug monitoring is generally unavailable. This study aimed to evaluate nevirapine concentrations in saliva using low-cost thin-layer chromatography (TLC) and nevirapine concentrations in plasma and saliva using high performance liquid chromatography (HPLC) methods; and to correlate nevirapine plasma concentrations to HIV treatment outcomes in Ugandan patients. Paired plasma and stimulated saliva samples were obtained from Ugandan, HIV-infected adults on nevirapine-based ART. Nevirapine concentrations were measured using a validated HPLC method and a novel TLC method. Plasma nevirapine concentrations HPLC were considered subtherapeutic. Negative/positive predictive values of different thresholds for subtherapeutic nevirapine concentrations in saliva were determined. Virologic testing and, if applicable, HIV drug resistance testing was performed. Median (interquartile range, IQR) age of 297 patients was 39.1 (32.8-45.2) years. Three hundred saliva and 287 plasma samples were available for analysis. Attempts failed to determine nevirapine saliva concentrations by TLC. Using HPLC, median (IQR) nevirapine concentrations in saliva and plasma were 3.40 (2.59-4.47) mg/L and 6.17 (4.79-7.96) mg/L, respectively. The mean (coefficient of variation,%) nevirapine saliva/plasma ratio was 0.58 (62%). A cut-off value of 1.60 mg/L nevirapine in saliva was associated with a negative/positive predictive value of 0.99/0.72 and a sensitivity/specificity of 87%/98% for predicting subtherapeutic nevirapine plasma concentrations, respectively. Only 5% (15/287) of patients had subtherapeutic nevirapine plasma concentrations, of which 3 patients had viral load results > 400 copies/mL. Patients with nevirapine concentrations in plasma 400 copies/mL). The low-cost TLC technique for monitoring nevirapine in saliva was unsuccessful but monitoring nevirapine saliva and plasma

  15. Development of Validated High-performance Thin-layer Chromatography Method for Simultaneous Determination of Quercetin and Kaempferol in Thespesia populnea.

    Science.gov (United States)

    Panchal, Hiteksha; Amin, Aeshna; Shah, Mamta

    2017-01-01

    Thespesia populnea L. (Family: Malvaceae) is a well-known medicinal plant distributed in tropical regions of the world and cultivated in South Gujarat and indicated to be useful in cutaneous affections, psoriasis, ringworm, and eczema. Bark and fruits are indicated in the diseases of skin, urethritis, and gonorrhea. The juice of fruits is employed in treating certain hepatic diseases. The plant is reported to contain flavonoids, quercetin, kaempferol, gossypetin, Kaempferol-3-monoglucoside, β-sitosterol, kaempferol-7-glucoside, and gossypol. T. populnea is a common component of many herbal and Ayurvedic formulation such as Kamilari and Liv-52. The present study aimed at developing validated and reliable high-performance thin layer chromatography (HPTLC) method for the analysis of quercetin and kaempferol simultaneously in T. populnea . The method employed thin-layer chromatography aluminum sheets precoated with silica gel as the stationary phase and toluene: ethyl acetate: formic acid (6:4:0.3 v/v/v) as the mobile phase, which gave compact bands of quercetin and kaempferol. Linear regression data for the calibration curves of standard quercetin and kaempferol showed a good linear relationship over a concentration range of 100-600 ng/spot and 500-3000 ng/spot with respect to the area and correlation coefficient (R2) was 0.9955 and 0.9967. The method was evaluated regarding accuracy, precision, selectivity, and robustness. Limits of detection and quantitation were recorded as 32.06 and 85.33 ng/spot and 74.055 and 243.72 ng/spot for quercetin and kaempferol, respectively. We concluded that this method employing HPTLC in the quantitative determination of quercetin and kaempferol is efficient, simple, accurate, and validated.

  16. Analysis of Bioactive Components of Oilseed Cakes by High-Performance Thin-Layer Chromatography-(Bioassay Combined with Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Sue-Siang Teh

    2015-03-01

    Full Text Available Hemp, flax and canola seed cakes are byproducts of the plant oil extraction industry that have not received much attention in terms of their potential use for human food instead of animal feed. Thus, the bioactivity profiling of these oilseed cakes is of interest. For their effect-directed analysis, planar chromatography was combined with several (bioassays, namely 2,2-diphenyl-1-picrylhydrazyl scavenging, acetylcholine esterase inhibition, planar yeast estrogen screen, antimicrobial Bacillus subtilis and Aliivibrio fischeri assays. The streamlined high-performance thin-layer chromatography (HPTLC-bioassay method allowed the discovery of previously unknown bioactive compounds present in these oilseed cake extracts. In contrast to target analysis, the direct link to the effective compounds allowed comprehensive information with regard to selected effects. HPTLC-electrospray ionization-mass spectrometry via the elution-head based TLC-MS Interface was used for a first characterization of the unknown effective compounds. The demonstrated bioactivity profiling on the feed/food intake side may guide the isolation of active compounds for production of functional food or for justified motivation of functional feed/food supplements.

  17. Lysergic acid amide as chemical marker for the total ergot alkaloids in rye flour - Determination by high-performance thin-layer chromatography-fluorescence detection.

    Science.gov (United States)

    Oellig, Claudia

    2017-07-21

    Ergot alkaloids are generally determined by high-performance liquid chromatography (HPLC) coupled to fluorescence detection (FLD) or mass selective detection, analyzing the individual compounds. However, fast and easy screening methods for the determination of the total ergot alkaloid content are more suitable, since for monitoring only the sum of the alkaloids is relevant. The herein presented screening uses lysergic acid amide (LSA) as chemical marker, formed from ergopeptine alkaloids, and ergometrine for the determination of the total ergot alkaloids in rye with high-performance thin-layer chromatography-fluorescence detection (HPTLC-FLD). An ammonium acetate buffered extraction step was followed by liquid-liquid partition for clean-up before the ergopeptine alkaloids were selectively transformed to LSA and analyzed by HPTLC-FLD on silica gel with isopropyl acetate/methanol/water/25% ammonium hydroxide solution (80:10:3.8:1.1, v/v/v/v) as the mobile phase. The enhanced native fluorescence of LSA and unaffected ergometrine was used for quantitation without any interfering matrix. Limits of detection and quantitation were 8 and 26μg LSA/kg rye, which enables the determination of the total ergot alkaloids far below the applied quality criterion limit for rye. Close to 100% recoveries for different rye flours at relevant spiking levels were obtained. Thus, reliable results were guaranteed, and the fast and efficient screening for the total ergot alkaloids in rye offers a rapid alternative to the HPLC analysis of the individual compounds. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Thin-layer and paper chromatography

    International Nuclear Information System (INIS)

    Sherma, J.

    1986-01-01

    This selective review covers the literature of thin-layer chromatography (TLC) and paper chromatography (PC) cited in Chemical Abstracts from December 5, 1983, through November 25, 1985, and Analytical Abstracts from November 1983 to November 1985. Also researched directly were the following important journals publishing papers on TLC and PC: the Journal of Chromatography (including its bibliography issues), Journal of High Resolution Chromatography and Chromatography Communications, Journal of Chromatographic Science, Chromatographia, Analytical Chemistry, JAOAC, and the special TLC issues of the Journal of Liquid Chromatography. Many of the inherent advantages of TLC that are obvious to workers familiar with high performance, quantitative theory and practice still are not appreciated adequately by the majority of people using chromatography. These include unrestricted access to the separation process; introducing magnetic, thermal, electrical, and other physical forces to improve resolution; high sample throughput; truly multidimensional separations; and the use of controlled multiple gradients. Many advantages of TLC relative to column chromatography were discussed in the Introductions to our 1982 and 1984 reviews of TLC in this Journal. No complete commercial robotics system specifically for TLC has been developed, but all necessary modules are available for such a system. The combination of robotics, with the continued development of theory, practice, and instrumentation will lead eventually to TLC systems that are unrivaled for speed, versatility, accuracy, precision, and sensitivity. 573 references

  19. Determination of caffeine, theobromine and theophylline in Mate beer and Mate soft drinks by high-performance thin-layer chromatography.

    Science.gov (United States)

    Oellig, Claudia; Schunck, Jacob; Schwack, Wolfgang

    2018-01-19

    Mate beer and Mate soft drinks are beverages produced from the dried leaves of Ilex paraguariensis (Yerba Mate). In Yerba Mate, the xanthine derivatives caffeine, theobromine and theophylline, also known as methylxanthines, are important active components. The presented method for the determination of caffeine, theobromine and theophylline in Mate beer and Mate soft drinks by high-performance thin-layer chromatography with ultraviolet detection (HPTLC-UV) offers a fully automated and sensitive determination of the three methylxanthines. Filtration of the samples was followed by degassing, dilution with acetonitrile in the case of Mate beers for protein precipitation, and centrifugation before the extracts were analyzed by HPTLC-UV on LiChrospher silica gel plates with fluorescence indicator and acetone/toluene/chloroform (4:3:3, v/v/v) as the mobile phase. For quantitation, the absorbance was scanned at 274nm. Limits of detection and quantitation were 1 and 4ng/zone, respectively, for caffeine, theobromine and theophylline. With recoveries close to 100% and low standard deviations reliable results were guaranteed. Experimental Mate beers as well as Mate beers and Mate soft drinks from the market were analyzed for their concentrations of methylxanthines. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Comparison and characterization of soybean and sunflower lecithins used for chocolate production by high-performance thin-layer chromatography with fluorescence detection and electrospray mass spectrometry.

    Science.gov (United States)

    Krüger, Stephanie; Bürmann, Laura; Morlock, Gertrud E

    2015-03-25

    The scarce availability of nongenetically modified soybeans on the world market represents a growing problem for food manufacturers. Hence, in this study the effects of substituting soybean with sunflower lecithin were investigated with regard to chocolate production. The glycerophospholipid pattern of the different lecithin samples was investigated by high-performance thin-layer chromatography fluorescence detection (HPTLC-FLD) and by HPTLC-positive ion electrospray ionization mass spectrometry (ESI(+)-MS) via the TLC-MS Interface and by scanning HPTLC-matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOFMS). Especially, the contents of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were of interest due to the influencing effects of these two glycerophospholipids on the rheological parameters of chocolate production. The lecithin substitution led to only slight differences in the rheological parameters of milk and dark chocolate. Limits of detection (LODs) and limits of quantification (LOQs) of seven glycerophospholipids were studied for three detection modes. Mean LODs ranged from 8 to 40 mg/kg for HPTLC-FLD and, using a single-quadrupole MS, from 10 to 280 mg/kg for HPTLC-ESI(+)-MS as well as from 15 to 310 mg/kg for HPTLC-FLD-ESI(+)-MS recorded after derivatization with the primuline reagent.

  1. Analysis of biodiesel and biodiesel-petrodiesel blends by high performance thin layer chromatography combined with easy ambient sonic-spray ionization mass spectrometry.

    Science.gov (United States)

    Eberlin, Livia S; Abdelnur, Patricia V; Passero, Alan; de Sa, Gilberto F; Daroda, Romeu J; de Souza, Vanderlea; Eberlin, Marcos N

    2009-08-01

    High performance thin layer chromatography (HPTLC) combined with on-spot detection and characterization via easy ambient sonic-spray ionization mass spectrometry (EASI-MS) is applied to the analysis of biodiesel (B100) and biodiesel-petrodiesel blends (BX). HPTLC provides chromatographic resolution of major components whereas EASI-MS allows on-spot characterization performed directly on the HPTLC surface at ambient conditions. Constituents (M) are detected by EASI-MS in a one component-one ion fashion as either [M + Na](+) or [M + H](+). For both B100 and BX samples, typical profiles of fatty acid methyl esters (FAME) detected as [FAME + Na](+) ions allow biodiesel typification. The spectrum of the petrodiesel spot displays a homologous series of protonated alkyl pyridines which are characteristic for petrofuels (natural markers). The spectrum for residual or admixture oil spots is characterized by sodiated triglycerides [TAG + Na](+). The application of HPTLC to analyze B100 and BX samples and its combination with EASI-MS for on-spot characterization and quality control is demonstrated.

  2. Algerian mint species:high performance thin layer chromatography quantitative determination of rosmarinic acid and in vitro inhibitory effects on linoleic acid peroxidation

    Institute of Scientific and Technical Information of China (English)

    Brahmi Fatiha; Madani Khodir; Stvigny Caroline; Chibane Mohamed; Duez Pierre

    2014-01-01

    Objective: To determine the quantitative paremeters of rosmarinic acid in Algerian mints,Mentha spicata L. (M. spicata), Mentha pulegium L. and Mentha rotundifolia (L.) Huds by high performance thin layer chromatography (HPTLC)-densitometric method and screen the effects of these plant extracts on linoleic acid peroxidation.Methods:The analyses were performed on HPTLC silica gel 60 F254 plates with chloroform:acetone: formic acid (75:16.5:8.5, v/v) as the mobile phase. Rosmarinic acid was determined in UV at 365 nm and fluorescence at λexc 325 nm with a 550 nm filter, respectively. The effects of plants extracts on linoleic acid peroxidation were measured by an indirect in vitro colorimetric method.Results:Chromatographic resolution permitted reliable quantification in both measurement modes and calibration curves were linear in a range of 150-1000 ng/spot. M. spicata was found to contain significantly higher concentrations of rosmarinic acid. The densitometric quantification allowed the analysis of many samples in a short time with reasonable precision (total precision for Mentha spp extracts, 5.1% and 5.8% for UV and fluorescence detection, respectively). The HPTLC data, allied to assays of linoleic acid peroxidation prevention, suggested the potential of M. spicata (52% Trolox®equivalents) as a natural source for inhibitors of lipid peroxidation.Conclusions:Densitometry can be used for routine determination and quality control of rosmarinic acid in herbal and formulations containing Mentha species.

  3. Detection and Estimation of alpha-Amyrin, beta-Sitosterol, Lupeol, and n-Triacontane in Two Medicinal Plants by High Performance Thin Layer Chromatography

    Directory of Open Access Journals (Sweden)

    Saikat S. Mallick

    2014-01-01

    Full Text Available A normal phase high performance thin layer chromatography (HPTLC method has been developed and validated for simultaneous estimation of four components, namely, alpha-amyrin, beta-sitosterol, lupeol, and n-triacontane from two medicinally important plants, Leptadenia reticulata Wight & Arn. and Pluchea lanceolata (DC. CB. Clarke. In Ayurveda, both plants have been reported to possess immunomodulatory activity. Chromatographic separation of the four components from the methanolic extracts of whole plant powders of Leptadenia reticulata Wight & Arn. and Pluchea lanceolata (DC. CB. Clarke. was performed on TLC aluminium plates precoated with silica gel 60F254 using a suitable mobile phase. The densitometric scanning was done after derivatization at λ = 580 nm for α-amyrin, β-sitosterol, and lupeol, and at 366 nm for n-triacontane. The developed HPTLC method has been validated and used for simultaneous quantitation of the four components from the methanolic extracts of whole plant powders of Leptadenia reticulata Wight & Arn. and Pluchea lanceolata (DC. CB. Clarke. The developed HPTLC method is simple, rapid, and precise and can be used for routine quality control.

  4. Algerian mint species: high performance thin layer chromatography quantitative determination of rosmarinic acid and in vitro inhibitory effects on linoleic acid peroxidation

    Directory of Open Access Journals (Sweden)

    Brahmi Fatiha

    2014-12-01

    Full Text Available Objective: To determine the quantitative paremeters of rosmarinic acid in Algerian mints, Mentha spicata L. (M. spicata, Mentha pulegium L. and Mentha rotundifolia (L. Huds by high performance thin layer chromatography (HPTLC-densitometric method and screen the effects of these plant extracts on linoleic acid peroxidation. Methods: The analyses were performed on HPTLC silica gel 60 F254 plates with chloroform: acetone: formic acid (75:16.5:8.5, v/v as the mobile phase. Rosmarinic acid was determined in UV at 365 nm and fluorescence at λexc 325 nm with a 550 nm filter, respectively. The effects of plants extracts on linoleic acid peroxidation were measured by an indirect in vitro colorimetric method. Results: Chromatographic resolution permitted reliable quantification in both measurement modes and calibration curves were linear in a range of 150-1 000 ng/spot. M. spicata was found to contain significantly higher concentrations of rosmarinic acid. The densitometric quantification allowed the analysis of many samples in a short time with reasonable precision (total precision for Mentha spp extracts, 5.1% and 5.8% for UV and fluorescence detection, respectively. The HPTLC data, allied to assays of linoleic acid peroxidation prevention, suggested the potential of M. spicata (52% Trolox® equivalents as a natural source for inhibitors of lipid peroxidation. Conclusions: Densitometry can be used for routine determination and quality control of rosmarinic acid in herbal and formulations containing Mentha species.

  5. Identification of Ginkgo biloba supplements adulteration using high performance thin layer chromatography and ultra high performance liquid chromatography-diode array detector-quadrupole time of flight-mass spectrometry.

    Science.gov (United States)

    Avula, Bharathi; Sagi, Satyanarayanaraju; Gafner, Stefan; Upton, Roy; Wang, Yan-Hong; Wang, Mei; Khan, Ikhlas A

    2015-10-01

    Ginkgo biloba is one of the most widely sold herbal supplements and medicines in the world. Its popularity stems from having a positive effect on memory and the circulatory system in clinical studies. As ginkgo popularity increased, non-proprietary extracts were introduced claiming to have a similar phytochemical profile as the clinically tested extracts. The standardized commercial extracts of G. biloba leaf used in ginkgo supplements contain not less than 6% sesquiterpene lactones and 24% flavonol glycosides. While sesquiterpene lactones are unique constituents of ginkgo leaf, the flavonol glycosides are found in many other botanical extracts. Being a high value botanical, low quality ginkgo extracts may be subjected to adulteration with flavonoids to meet the requirement of 24% flavonol glycosides. Chemical analysis by ultra high performance liquid chromatography-mass spectrometry revealed that adulteration of ginkgo leaf extracts in many of these products is common, the naturally flavonol glycoside-rich extract being spiked with pure flavonoids or extracts made from another flavonoid-rich material, such as the fruit/flower of Japanese sophora (Styphnolobium japonicum), which also contains the isoflavone genistein. Recently, genistein has been proposed as an analytical marker for the detection of adulteration of ginkgo extracts with S. japonicum. This study confirms that botanically authenticated G. biloba leaf and extracts made therefrom do not contain genistein, and the presence of which even in trace amounts is suggestive of adulteration. In addition to the mass spectrometric approach, a high performance thin layer chromatography method was developed as a fast and economic method for chemical fingerprint analysis of ginkgo samples.

  6. Effect-directed fingerprints of 77 botanical extracts via a generic high-performance thin-layer chromatography method combined with assays and mass spectrometry.

    Science.gov (United States)

    Krüger, S; Hüsken, L; Fornasari, R; Scainelli, I; Morlock, G E

    2017-12-22

    Quantitative effect-directed profiles of 77 industrially and freshly extracted botanicals like herbs, spices, vegetables and fruits, widely used as food ingredients, dietary supplements or traditional medicine, gave relevant information on their quality. It allows the assessment of food, dietary supplements and phytomedicines with regard to potential health-promoting activities. In contrary to sum parameter assays and targeted analysis, chromatography combined with effect-directed analysis allows fast assignment of single active compounds and evaluation of their contribution to the overall activity, originating from a food or botanical sample. High-performance thin-layer chromatography was hyphenated with UV/Vis/FLD detection and effect-directed analysis, using the 2,2-diphenyl-1-picrylhydrazyl radical, Gram-negative Aliivibrio fischeri, Gram-positive Bacillus subtilis, acetylcholinesterase and tyrosinase assays. Bioactive compounds of interest were eluted using an elution head-based interface and further characterized by electrospray ionization (high-resolution) mass spectrometry. This highly streamlined workflow resulted in a hyphenated HPTLC-UV/Vis/FLD-EDA-ESI + /ESI - -(HR)MS method. The excellent quantification power of the method was shown on three compounds. For rosmarinic acid, contents ranged from 4.5mg/g (rooibos) to 32.6mg/g (rosemary), for kaempferol-3-glucoside from 0.6mg/g (caraway) to 4.4mg/g (wine leaves), and for quercetin-3-glucoside from 1.1mg/g (hawthorn leaves) to 17.7mg/g (thyme). Three mean repeatabilities (%RSD) over 18 quantifications for the three compounds were ≤2.2% and the mean intermediate precision over three different days (%RSD, n=3) was 5.2%. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Anti-inflammatory activity and qualitative analysis of different extracts of Maytenus obscura (A. Rich.) Cuf. by high performance thin layer chromatography method

    Institute of Scientific and Technical Information of China (English)

    Mohamed F. Alajmi; Perwez Alam

    2014-01-01

    Objective: To perform aqueous ethanol soluble fraction (AESF) and dichloromethane extract of aerial parts of Maytenus obscura (A. Rich.) Cuf. using high performance thin layer chromatography (HPTLC) and to test anti-inflammatory activity of these extracts.Methods:HPTLC studies were carried out using CAMAG HPTLC system equipped with Linomat IV applicator, TLC scanner 3, Reprostar 3, CAMAG ADC 2 and WIN CATS-4 software were used. The anti-inflammatory activity was tested by injecting different groups of rats (6 each) with formalin in hind paw and measuring the edema volume before and 1 h later formalin injection. Control group received saline i.p. The extracts treatment was injected i.p. in doses of 100 and 200 mg/kg 1 h before formalin administration. Indomethacin (30 mg/kg) was used as standard.Results:The results of preliminary phytochemical studies confirmed the presence of protein, lipid, carbohydrate, phenol, flavonoid, saponin, triterpenoid, alkaloid and anthraquinone in both extracts. Chromatography was performed on glass-backed silica gel 60 F254 HPTLC plates with the green solvents toluene: ethyacetate: glacial acetic acid (5:3:0.2, v/v/v) as mobile phase. HPTLC finger printing of AESF revealed major eight peaks with Rf values in the range of 0.28 to 0.80 and the dichloromethane revealed major 11 peaks with Rf values in the range of 0.12 to 0.76. The purity of sample was confirmed by comparing the absorption spectra at start, middle and end position of the band. Treatment of rats (i.p.) with AESF and dichloromethane in doses of 100 and 200 mg/kg inhibited singnificantly (P<0.05, n=6) formalin-induced inflammation by 50%, 55.9%, 45.5%, and 51.4%, respectively.Conclusions:HPTLC finger printing of AESF and dichloromethane of Maytenus obscura revealed eight major spots for alcoholic extracts and nine major spots for dichloromethane extracts. These HPTLC profiles may be of great usefulness in the quality control of herbal products containing these extracts. The

  8. Chemical variability along the value chains of turmeric (Curcuma longa): a comparison of nuclear magnetic resonance spectroscopy and high performance thin layer chromatography.

    Science.gov (United States)

    Booker, Anthony; Frommenwiler, Debora; Johnston, Deborah; Umealajekwu, Chinenye; Reich, Eike; Heinrich, Michael

    2014-03-14

    Herbal medicine value chains have generally been overlooked compared with food commodities. Not surprisingly, revenue generation tends to be weighted towards the end of the chain and consequently the farmers and producers are the lowest paid beneficiaries. Value chains have an impact both on the livelihood of producers and on the composition and quality of products commonly sold locally and globally and consequently on the consumers. In order to understand the impact of value chains on the composition of products, we studied the production conditions for turmeric (Curcuma longa) and the metabolomic composition of products derived from it. We aimed at integrating these two components in order to gain a better understanding of the effect of different value chains on the livelihoods of some producers. This interdisciplinary project uses a mixed methods approach. Case studies were undertaken on two separate sites in India. Data was initially gathered on herbal medicine value chains by means of semi-structured interviews and non-participant observations. Samples were collected from locations in India, Europe and the USA and analysed using (1)H NMR spectroscopy coupled with multivariate analysis software and with high performance thin layer chromatography (HPTLC). We investigate medicinal plant value chains and interpret the impact different value chains have on some aspects of the livelihoods of producers in India and, for the first time, analytically assess the chemical variability and quality implications that different value chains may have on the products available to end users in Europe. There are benefits to farmers that belonged to an integrated chain and the resulting products were subject to a higher standard of processing and storage. By using analytical methods, including HPTLC and (1)H NMR spectroscopy, it has been possible to correlate some variations in product composition for selected producers and identify strengths and weaknesses of some types of value

  9. The offline combination of thin-layer chromatography and high-performance liquid chromatography with diode array detection and micrOTOF-Q mass spectrometry for the separation and identification of spinochromes from sea urchin (Strongylocentrotus droebachiensis) shells.

    Science.gov (United States)

    Shikov, Alexander N; Ossipov, Vladimir I; Martiskainen, Olli; Pozharitskaya, Olga N; Ivanova, Svetlana A; Makarov, Valery G

    2011-12-16

    Thin-layer chromatography (TLC) with off-line high-performance liquid chromatography coupled to diode array detection and micrOTOF-Q mass spectrometry (HPLC-DAD-MS) resulted in the successful fractionation, separation and identification of spinochrome pigments from sea urchin (Strongylocentrotus droebachiensis) shells. Two fractions of pigments were separated by TLC and eluted with methanol using a TLC-MS interface. HPLC-DAD-MS analysis of the fractions indicated the presence of six sea urchin pigments: spinochrome monomers B and D, three spinochrome dimers (anhydroethylidene-6,6'-bis(2,3,7-trihydroxynaphthazarin) and its isomer and ethylidene-6,6'-bis(2,3,7-trihydroxynaphthazarin)), and one pigment that was preliminary identified as a spinochrome dimer with the structural formula C(22)H(16)O(16). Copyright © 2011 Elsevier B.V. All rights reserved.

  10. Determination of Four Major Saponins in Skin and Endosperm of Seeds of Horse Chestnut (Aesculus Hippocastanum L.) Using High Performance Liquid Chromatography with Positive Confirmation by Thin Layer Chromatography.

    Science.gov (United States)

    Abudayeh, Zead Helmi Mahmoud; Al Azzam, Khaldun Mohammad; Naddaf, Ahmad; Karpiuk, Uliana Vladimirovna; Kislichenko, Viktoria Sergeevna

    2015-11-01

    To separate and quantify four major saponins in the extracts of the skin and the endosperm of seeds of horse chestnut (Aesculus hippocastanum L.) using ultrasonic solvent extraction followed by a high performance liquid chromatography-diode array detector (HPLC-DAD) with positive confirmation by thin layer chromatography (TLC). The saponins: escin Ia, escin Ib, isoescin Ia and isoescin Ib were extracted using ultrasonic extraction method. The optimized extraction conditions were: 70% methanol as extraction solvent, 80 °C as extraction temperature, and the extraction time was achieved in 4 hours. The HPLC conditions used: Zorbax SB-ODS-(150 mm × 2.1 mm, 3 μm) column, acetonitrile and 0.10% phosphoric acid solution (39:61 v/v) as mobile phase, flow rate was 0.5 mL min(-1) at 210 nm and 230 nm detection. The injection volume was 10 μL, and the separation was carried out isothermally at 30 °C in a heated chamber. The results indicated that the developed HPLC method is simple, sensitive and reliable. Moreover, the content of escins in seeds decreased by more than 30% in endosperm and by more than 40% in skin upon storage for two years. This assay can be readily utilized as a quality control method for horse chestnut and other related medicinal plants.

  11. Correlation between dried blood spot thin layer chromatography and plasma high performance liquid chromatography of leucine/isoleucine levels among Filipino patients with maple syrup urine disease (MSUD) seen at the Institute of Human Genetics, National Institutes of Health

    International Nuclear Information System (INIS)

    Yaplito-Lee, Joy; Chiong, Mary Anne D.; Rana, Michelle D.; Rama, Kahlil Izza D.; David-Padilla, Carmencita; Cavan, Barbra Charina; Cordero, Cynthia P.

    2008-01-01

    Management of patients with maple syrup urine disease (MSUD) includes a low protein diet, supplemented with special formulas and constant monitoring of branched chain amino acids (BCAA). The gold standard for monitoring BCAA is plasma amino acid analysis using high performance liquid chromatography (HPLC). In a developing country like the Philippines, however, the cost of this test is prohibitive to the majority of the patients. In our center, dried blood spot leucine/isoleucine (leu/ile) levels analysed by thin layer chromatography (TLC) is often used to diagnose and monitor these patients. This study was done to determine the correlation of leu/ile levels using the two methods (TLC and HPLC). A total of 46 MSUD patients were referred to the Biochemical Genetics Laboratory of the Institute of Human Genetics (IHG) from July 2001 to January 2004. Thirty five samples were obtained from 18 of these patients (some patients were seen at IHG more than once), and paired determinations of plasma amino acid using TLC and HPLC were made. The remaining samples were either hemolyzed or were not analyzed. The correlation coefficient [rho denoted as ρ] was estimated at a 95% confidence level using the Fisher's Z transformation. Of the 18 patients, 12 were males. The youngest was 1 day old and the oldest was 5 years old. The majority had the classical type of MSUD and dietary protein was restricted to between 0.6 gram/kg/day to 1 gram/kg/day of natural protein. Using the first pairs of observation for these 18 patients, the correlation coefficient was 0.76 (95% C1:0.462 to 0.907). This suggest a strong correlation between the two methods. It is recommended that further studies be done to determine the potential of the dried blood spot leu/ile level by TLC as an alternative method that can be used in the diagnosis and monitoring of MSUD patients especially in a developing country. (Author)

  12. Effects of Different Levels of Echinostoma caproni Miracidial Dose on Glucose and Maltose Composition of Biomphalaria glabrata Snails as Determined by High Performance Thin-Layer Chromatography-Densitometry

    Directory of Open Access Journals (Sweden)

    Dolcie DeGrandchamp

    2015-04-01

    Full Text Available The effects of 5, 25, and 40 Echinostoma caproni miracidia on the sugar content of young adult and mature adult Biomphalaria glabrata were studied using high performance thin layer chromatography (HPTLC-densitometry. Analysis was done on the snail’s digestive gland gonad complex (DGG at two and four weeks postmiracidial exposure. The sugars were extracted from the DGG using 70% ethanol and analyzed on silica gel HPTLC plates with a preadsorbent zone using 1-butanol-glacial acetic acid-diethyl ether-deionized water (27:18:5:3 mobile phase. The separated bands were then detected using alpha-naphthol-sulfuric reagent and quantified by densitometry at 515 nm. Significant differences were found in the maltose content between two and four weeks post exposure for both age groups. Additionally, significantly lower maltose and glucose levels were observed in the high exposure groups of both ages.

  13. Loading Effects on Resolution in Thin Layer Chromatography and ...

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 10; Issue 11. Loading Effects on Resolution in Thin Layer Chromatography and Paper Chromatography. K Girigowda V H Mulimani. Classroom Volume 10 Issue 11 November 2005 pp 79-84 ...

  14. High-performance thin-layer chromatography analysis of steviol glycosides in Stevia formulations and sugar-free food products, and benchmarking with (ultra) high-performance liquid chromatography.

    Science.gov (United States)

    Morlock, Gertrud E; Meyer, Stephanie; Zimmermann, Benno F; Roussel, Jean-Marc

    2014-07-11

    A high-performance TLC (HPTLC) method was newly developed and validated for analysis of 7 steviol glycosides in 6 different types of food and Stevia formulations. After a minimized one-step sample preparation, 21 samples were developed in parallel, allowing an effective food screening. Depending on the sample application volume, the method was suited to analyze food sample concentrations in the mg/kg range. LOQs of stevioside in natural yoghurt matrix spiked at 0.02, 0.13 and 0.2% were determined by the calibration curve method to be 12ng/band (peak height). ANOVA was successfully passed to prove data homogeneity in the working range (30-600ng/band). The accuracy (recovery tolerance limit, 92-120%), repeatability (3.1-5.4%) and intermediate precision (4.0-8.4%) were determined for stevioside in milk-based matrix including sample preparation and recovery rates at 3 different concentration levels. For the first time, the recording of HPTLC-ESI-MS spectra via the TLC-MS Interface was demonstrated for rebaudioside A. HPTLC contents for rebaudioside A were compared with results of two (U)HPLC methods. The running costs and analysis time of the three different methods were discussed in detail with regard to screening of food products. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Thin-Layer and Paper Chromatography.

    Science.gov (United States)

    Sherma, Joseph; Fried, Bernard

    1984-01-01

    Reviews literature on chromatography examining: books, reviews, student experiments; chromatographic systems, techniques, apparatus; detecting and identification of separated zones; preparative chromatography and radiochromatography; and applications related to specific materials (such as acids, alcohols, amino acids, antibiotics, enzymes, dyes,…

  16. Pre-staining thin layer chromatography method for amino acid ...

    African Journals Online (AJOL)

    Jane

    2010-12-13

    Dec 13, 2010 ... inexpensive and the results obtained were clean and reproducible. However, it is suitable for the high throughput screening of amino acid-producing strains. Key words: Thin layer chromatography, pre-staining, amino acid detection. INTRODUCTION. Several analytical techniques have been often used for.

  17. Thin-layer chromatography - liquid chromatography, an ideal supplement demonstrated by the separation of lanthanoids

    International Nuclear Information System (INIS)

    Specker, H.; Hufnagel, A.

    1984-01-01

    All lanthanoids have been separated by thin-layer chromatography (TLC) with short retention times by using a mixture of ether/tetrahydrofurane (THF)/bis-(2-ethylhexyl)-phosphate (HDEHP)/nitric acid. The eluent was empirically tested by synergistic effects. The results have been transferred to high-performance liquid chromatography /HPLC). It was possible to use the same eluent in TLC and HPLC both for the analytical separation of all lanthanoids and for the separation of fission products. The experimental experience gained in HPLC could be applied to the pre-concentration of isotopes in TLC. Both methods excellently supplemented each other in the separation of lanthanoids. (orig.) [de

  18. Application of thin-layer chromatography in radiochemistry

    International Nuclear Information System (INIS)

    Maki, Yasuyuki; Murakami, Yukio.

    1976-01-01

    In relation to the experimental procedures of thin-layer chromatography (TLC) in radiochemistry, the authors explained the preparation and development of radioactive test solutions, the methods of detection by autoradiography of isolated spots and by the calculation of measuring apparatus, and the identification of isolated spots. Next they outlined the carrier-free isolation and purification of nuclides, the quantification in combination with γ-ray spectrum, confirmation of the purity of RI-labeled medical supplies, their application to RI generator, thin-layer electrophoresis, in which electrophoresis and TLC were combined, and the application of this electrophoresis to isolation in recoil chemistry and to analysis and identification in carrier-free chemistry. (Kanao, K.)

  19. Development and validation of a simple high performance thin layer chromatography method combined with direct 1,1-diphenyl-2-picrylhydrazyl assay to quantify free radical scavenging activity in wine.

    Science.gov (United States)

    Agatonovic-Kustrin, Snezana; Morton, David W; Yusof, Ahmad P

    2016-04-15

    The aim of this study was to: (a) develop a simple, high performance thin layer chromatographic (HPTLC) method combined with direct 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay to rapidly assess and compare free radical scavenging activity or anti-oxidant activity for major classes of polyphenolics present in wines; and (b) to investigate relationship between free radical scavenging activity to the total polyphenolic content (TPC) and total antioxidant capacity (TAC) in the wine samples. The most potent free radical scavengers that we tested for in the wine samples were found to be resveratrol (polyphenolic non-flavonoid) and rutin (flavonoid), while polyphenolic acids (caffeic acid and gallic acid) although present in all wine samples were found to be less potent free radical scavengers. Therefore, the total antioxidant capacity was mostly affected by the presence of resveratrol and rutin, while total polyphenolic content was mostly influenced by the presence of the less potent free radical scavengers gallic and caffeic acids. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Estimation of boswellic acids from market formulations of Boswellia serrata extract and 11-keto beta-boswellic acid in human plasma by high-performance thin-layer chromatography.

    Science.gov (United States)

    Shah, Shailesh A; Rathod, Ishwarsinh S; Suhagia, Bhanubhai N; Patel, Dharmesh A; Parmar, Vijay K; Shah, Bharat K; Vaishnavi, Vikas M

    2007-04-01

    A rapid and sensitive high-performance thin-layer chromatographic (HPTLC) method was developed and validated for the quantitative estimation of boswellic acids in formulation containing Boswellia serrata extract (BSE) and 11-keto beta-boswellic acid in human plasma. Simple extraction method was used for isolation of boswellic acid from formulation sample and acidified plasma sample. The isolated samples were chromatographed on silica gel 60F(254)-TLC plates, developed using ternary-solvent system (hexane-chloroform-methanol, 5:5:0.5, v/v) and scanned at 260 nm. The linearity range for 11-KBA spiked in 1 ml of plasma was 29.15-145.75 ng with average recovery of 91.66%. The limit of detection and limit of quantification for 11-KBA in human plasma were found to be 8.75 ng/ml and 29.15 ng/ml. The developed method was successfully applied for the assay of market formulations containing BSE and to determine plasma level of 11-keto beta-boswellic acid in a clinical pilot study.

  1. A Simple Thin Layer Chromatography Method for Separation of Selected Natural Steroid Hormones

    International Nuclear Information System (INIS)

    Nowakowska, J.; Rudnicka-Litka, K.; Ciura, K.; Pikul, P.; Piotrowicz, J.

    2015-01-01

    Chromatographic properties of seven steroids: estrogens (β-estradiol and estrone), androgens (testosterone, methyltestosterone, trans-androsterone), progesterone and cholesterol have been studied by planar chromatography with usage of High Performance Thin Layer Chromatography (HPTLC) and Thin Layer Chromatography (TLC) plates. Normal, reversed and cyano-bonded silica stationary phases were tested with five binary mobile phases (acetonitrile-water, acetonitrile-DMSO, acetonitrile-methanol, acetone-petroleum ether, acetone-water) in which the concentration of organic modifier varied from 0 to 100 % (v/v). This study reports the optimization of steroid hormones separation. Principal Component Analysis (PCA) based on calculated molecular descriptors quantitatively differentiating solutes was performed in order to investigate the similarity and dissimilarity between tested compounds. The separation abilities of mobile and stationary phases were compared based on separation factor α. Chromatographic retention data and possible retention mechanisms also were discussed. (author)

  2. Robotic thin layer chromatography instrument for synthetic chemistry

    International Nuclear Information System (INIS)

    Corkan, L.A.; Haynes, E.; Kline, S.; Lindsey, J.S.

    1991-01-01

    We have constructed a second generation instrument for performing automated thin layer chromatography (TLC), The TLC instrument Consists of four dedicated stations for (1) plate dispensing, (2) sample application, (3) plate development, and (4) densitometry. A robot is used to move TLC plates among stations. The TLC instrument functions either as a stand-alone unit or as one analytical module in a robotic workstation for synthetic chemistry. An integrated hardware and software architecture enables automatic TLC analysis of samples produced concurrently from synthetic reactions in progress on the workstation. The combination of fixed automation and robotics gives a throughput of 12 TLC samples per hour. From these results a blueprint has emerged for an advanced automated TLC instrument with far greater throughput and analytical capabilities

  3. A simple and rapid method for the determination of taxol produced by fungal endophytes from medicinal plants using high performance thin layer chromatography%高效薄层色谱法快速测定由药用植物内生真菌产生的紫杉醇

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Taxol is an important anticancer drug used widely in the clinical field. In this study, some endophytic fungi were isolated from selected medicinal plants, and were screened for their potential in the production of taxol, using a rapid separation technique of high performance thin layer chromatography ( HPTLC ). Of the 20 screened fungi, only 13 fungal species produced taxol in the artificial culture medium.The results of HPTLC showed that the 13 fungal species had identical ultraviolet (UV) characteristics, positive reactivity with a spray reagent, yielding a blue spot, which turned to dark gray after 24 hours, and had Rf values identical to that of the authentic taxol. The amount of taxol was also quantified by comparing the peak area and the peak height of the fungal samples with those of authentic taxol.

  4. High-performance thin-layer chromatography linked with (bio)assays and mass spectrometry - a suited method for discovery and quantification of bioactive components? Exemplarily shown for turmeric and milk thistle extracts.

    Science.gov (United States)

    Taha, Mahmoud N; Krawinkel, Michael B; Morlock, Gertrud E

    2015-05-15

    Extraction parameters, chemical fingerprint, and the single compounds' activity levels were considered for the selection of active botanicals. For an initial survey, the total bioactivity (i.e., total reducing capacity, total flavonoids contents and free radical scavenging capacity) of 21 aqueous and 21 ethanolic plant extracts was investigated. Ethanolic extracts showed a higher yield and were further analyzed by HPTLC in detail to obtain fingerprints of single flavonoids and further bioactive components. Exemplarily shown for turmeric (Curcuma longa) and milk thistle (Silybum marianum), effect-directed analysis (EDA) was performed using three selected (bio)assays, the Aliivibrio fischeri bioassay, the Bacillus subtilis bioassay and the 2,2-diphenyl-1-picrylhydrazyl (DPPH*) assay. As a proof of principle, the bioactive components found in the extracts were confirmed by HPTLC-MS. Bioassays in combination with planar chromatography directly linked to the known, single effective compounds like curcumin and silibinin. However, also some unknown bioactive components were discovered and exemplarily characterized, which demonstrated the strength of this kind of EDA. HPTLC-UV/Vis/FLD-EDA-MS could become a useful tool for selection of active botanicals and for the activity profiling of the active ingredients therein. The flexibility in effect-directed detections allows a comprehensive survey of effective ingredients in samples. This streamlined methodology comprised a non-targeted, effect-directed screening first, followed by a highly targeted characterization of the discovered bioactive compounds. HPTLC-EDA-MS can also be recommended for bioactivity profiling of food on the food intake side, as not only effective phytochemicals, but also unknown bioactive degradation products during food processing or contamination products or residues or metabolites can be detected. Thus, an efficient survey on potential food intake effects on wellness could be obtained. Having performed

  5. Normal and Reversed-Phase Thin Layer Chromatography of Green Leaf Extracts

    Science.gov (United States)

    Sjursnes, Birte Johanne; Kvittingen, Lise; Schmid, Rudolf

    2015-01-01

    Introductory experiments of chromatography are often conducted by separating colored samples, such as inks, dyes, and plant extracts, using filter paper, chalk, or thin layer chromatography (TLC) plates with various solvent systems. Many simple experiments have been reported. The relationship between normal chromatography and reversed-phase…

  6. Thin-layer chromatography of radioactively labelled cholesterol and precursors from biological material

    International Nuclear Information System (INIS)

    Pill, J.; Aufenanger, J.; Stegmeier, K.; Schmidt, F.H.; Mueller, D.; Boehringer Mannheim G.m.b.H.

    1987-01-01

    The investigation methods of the action of xenobiotics on sterol biosynthesis from 14 C-acetate in rat hepatocyte cultures can be developed, with regard to extraction using Extrelut and the separation of the sterol pattern by thin-layer chromatography, in such a way that they are suitable for wider application, e.g., screening. Good visualisation and recognition of changes in the sterol pattern are possible using autoradiography of the thin-layer chromatogram. (orig.)

  7. Development of High Performance Thin Layer Chromatography for ...

    African Journals Online (AJOL)

    are likely to be found on the market, especially in developing countries. ... was a deuterium lamp emitting a continuous UV spectrum in the range 200– ... disoproxil fumarate. Table 1. Summary of response function of lamivudine and tenofovir ...

  8. High-performance thin-layer chromatography (HPTLC)

    National Research Council Canada - National Science Library

    Srivastava, ManMohan

    2011-01-01

    ... on HPTLC fundamentals: principle, theory, understanding; instrumentation: implementation, optimization, validation, automation and qualitative and quantitative analysis; applications: phytochemical analysis, biomedical analysis, herbal drug quantification, analytical analysis, finger print analysis and potential for hyphenation: HPTLC future to combinat...

  9. Thin-layer chromatography can resolve phosphotyrosine, phosphoserine, and phosphothreonine in a protein hydrolyzate

    International Nuclear Information System (INIS)

    Neufeld, E.; Goren, H.J.; Boland, D.

    1989-01-01

    A solution of propionic acid, 1 M ammonium hydroxide, and isopropyl alcohol (45/17.5/17.5, v/v) was the ascending solvent in the separation of phosphotyrosine, phosphothreonine, and phosphoserine by thin-layer chromatography. The immobile phase was cellulose. The relative migrations were 0.44, 0.38, and 0.2, respectively. A previously described thin-layer system consisting of isobutyric acid and 0.5 M ammonium hydroxide (50/30, v/v) gave very similar relative migrations. To determine the usefulness of thin-layer chromatography in phosphoamino acid analysis, the propionic acid/ammonium hydroxide/isopropyl alcohol solution was used to characterize phosphorylated residues in a plasma membrane protein which is a substrate for the insulin receptor kinase, in insulin receptor phosphorylated histone H2B, and in an in vivo phosphorylated 90000-Da protein from IM9 cells. 32 P-labeled proteins were separated by dodecyl sulfate-gel electrophoresis, digested with trypsin, and then hydrolyzed with 6 N HCl, 2 h, 110 degrees C. Following thin-layer chromatography of the hydrolyzates and autoradiography, phosphotyrosine was detected in insulin receptor substrates, and phosphoserine and phosphothreonine were found in the in vivo-phosphorylated protein. This study supports previous reports about the practicality of thin-layer chromatography in phosphoamino acid analysis and it demonstrates that a propionic acid, ammonium hydroxide, isoprophyl alcohol solution may be a useful ascending solvent mixture for this purpose

  10. A simplified method for rapid quantification of intracellular nucleoside triphosphates by one-dimensional thin-layer chromatography

    DEFF Research Database (Denmark)

    Jendresen, Christian Bille; Kilstrup, Mogens; Martinussen, Jan

    2011-01-01

    -pyrophosphate (PRPP), and inorganic pyrophosphate (PPi) in cell extracts. The method uses one-dimensional thin-layer chromatography (TLC) and radiolabeled biological samples. Nucleotides are resolved at the level of ionic charge in an optimized acidic ammonium formate and chloride solvent, permitting...... quantification of NTPs. The method is significantly simpler and faster than both current two-dimensional methods and high-performance liquid chromatography (HPLC)-based procedures, allowing a higher throughput while common sources of inaccuracies and technical problems are avoided. For determination of PPi...

  11. High-performance thin-layer chromatographic fingerprints of isoflavonoids for distinguishing between Radix Puerariae Lobate and Radix Puerariae Thomsonii.

    Science.gov (United States)

    Chen, Si-Bao; Liu, He-Ping; Tian, Run-Tao; Yang, Da-Jian; Chen, Shi-Lin; Xu, Hong-Xi; Chan, Albert S C; Xie, Pei-Shan

    2006-07-14

    The roots of Pueraria lobata (Wild.) Ohwi and Pueraria thomsonii Benth have been officially recorded in all editions of Chinese Pharmacopoeia under the same monograph 'Gegen' (Radix Puerariae, RP). However, in its 2005 edition, the two species were separated into both individual monographs, namely 'Gegen' (Radix Puerariae Lobatae, RPL) and 'Fenge' (Radix Puerariae Thomsonii, RPT), respectively, due to their obvious content discrepancy of puerarin, the major active constituent. In present paper, the fingerprint of high-performance thin-layer chromatography (HPTLC) combining digital scanning profiling was developed to identify and distinguish the both species in detail. The unique properties of the HPTLC fingerprints were validated by analyzing ten batches of Pueraria lobata and P. thomsonii samples, respectively. The common pattern of the HPTLC images of the roots of Pueraria spp. and the respective different ratios of the chemical distribution can directly discern the two species. The corresponding digital scanning profiles provided an easy way for quantifiable comparison among the samples. Obvious difference in ingredient content and HPTLC patterns of the two species questioned their bio-equivalence and explained that recording both species separately in the current edition of Chinese Pharmacopoeia (2005 edition) is reasonable due to not only the content of major constituent, puerarin, but also the peak-to-peak distribution in the fingerprint and integration value of the total components. Furthermore, the HPTLC fingerprint is also suitable for rapid and simple authentication and comparison of the subtle difference among samples with identical plant resource but different geographic locations.

  12. Indirect fluorometric detection techniques on thin layer chromatography and effect of ultrasound on gel electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Yinfa, Ma.

    1990-12-10

    Thin-layer chromatography (TLC) is a broadly applicable separation technique. It offers many advantages over high performance liquid chromatography (HPLC), such as easily adapted for two-dimensional separation, for whole-column'' detection and for handling multiple samples, etc. However, due to its draggy development of detection techniques comparing with HPLC, TLC has not received the attention it deserves. Therefore, exploring new detection techniques is very important to the development of TLC. It is the principal of this dissertation to present a new detection method for TLC -- indirect fluorometric detection method. This detection technique is universal sensitive, nondestructive, and simple. This will be described in detail from Sections 1 through Section 5. Section 1 and 3 describe the indirect fluorometric detection of anions and nonelectrolytes in TLC. In Section 2, a detection method for cations based on fluorescence quenching of ethidium bromide is presented. In Section 4, a simple and interesting TLC experiment is designed, three different fluorescence detection principles are used for the determination of caffeine, saccharin and sodium benzoate in beverages. A laser-based indirect fluorometric detection technique in TLC is developed in Section 5. Section 6 is totally different from Sections 1 through 5. An ultrasonic effect on the separation of DNA fragments in agarose gel electrophoresis is investigated. 262 refs.

  13. Interaction between cholesterol and non-ionic surfactants studied by thin-layer chromatography

    Czech Academy of Sciences Publication Activity Database

    Forgács, E.; Cserháti, T.; Farkas, O.; Eckhardt, Adam; Mikšík, Ivan; Deyl, Zdeněk

    2004-01-01

    Roč. 27, č. 13 (2004), s. 1981-1992 ISSN 1082-6076 Grant - others:CZ-HU(CZ) Cooperation program Institutional research plan: CEZ:AV0Z5011922 Keywords : cholesterol * non-ionic surfactant * thin - layer chromatography Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 0.836, year: 2004

  14. Sensitive Thin-Layer Chromatography Detection of Boronic Acids Using Alizarin

    NARCIS (Netherlands)

    Duval, F.L.; Beek, van T.A.; Zuilhof, H.

    2012-01-01

    A new method for the selective and sensitive detection of boronic acids on thin-layer chromatography plates is described. The plate is briefly dipped in an alizarin solution, allowed to dry in ambient air, and observed under 366 nm light. Alizarin emits a bright yellow fluorescence only in the

  15. Thin-Layer Chromatography/Desorption Atmospheric Pressure Photoionization Orbitrap Mass Spectrometry of Lipids

    Czech Academy of Sciences Publication Activity Database

    Rejšek, Jan; Vrkoslav, Vladimír; Vaikkinen, A.; Haapala, M.; Kauppila, T. J.; Kostiainen, R.; Cvačka, Josef

    2016-01-01

    Roč. 88, č. 24 (2016), s. 12279-12286 ISSN 0003-2700 R&D Projects: GA ČR GAP206/12/0750 Institutional support: RVO:61388963 Keywords : desorption atmospheric pressure photoionization * thin-layer chromatography * lipids Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 6.320, year: 2016

  16. Phospholipids, Dietary Supplements, and Chicken Eggs: An Inquiry-Based Exercise Using Thin-Layer Chromatography

    Science.gov (United States)

    Potteiger, Sara E.; Belanger, Julie M.

    2015-01-01

    This inquiry-based experiment is designed for organic or biochemistry undergraduate students to deduce the identity of phospholipids extracted from chicken eggs and dietary supplements. This is achieved using thin-layer chromatography (TLC) data, a series of guided questions of increasing complexity, and provided relative retention factor (Rf)…

  17. Possible artefacts in thin layer chromatography of tritium-labelled hydrocortisone

    International Nuclear Information System (INIS)

    Sofronie, E.

    1982-12-01

    Artefacts appearing in thin layer chromatography of tritium labelled hydrocortisone are reported. Evidences are presented that these artefacts cause misleading results concerning radiocheemical purity determiniation. Finally, it is reported a rapid and efficient chromatographic technique allowing the elimination of these artefacts and obtaining of an accurate value for radiochemical purity. (author)

  18. Bibliography of paper and thin-layer chromatography 1970-1973 and survey of applications

    International Nuclear Information System (INIS)

    Macek, K.; Hais, I.M.; Kopecky, J.; Schwarz, V.; Gasparic, J.; Churacek, J.

    1976-01-01

    The present volume covers developments in both paper and thin-layer chromatography from 1970 to the middle of 1973 and is a continuation of the previous four volumes, published in 1960, 1962, 1968 and 1972. An author index is given

  19. Peak broadening in paper chromatography and related techniques : III. Peak broadening in thin-layer chromatography on cellulose powder

    NARCIS (Netherlands)

    Ligny, C.L. de; Remijnse, A.G.

    1968-01-01

    The mechanism of peak broadening in thin-layer chromatography on cellulose powder was investigated by comparing the peak widths obtained in chromatography with those caused only by diffusion in the cellulose powder, for a set of amino acids of widely differing RF values and six kinds of cellulose

  20. Use of low volatility mobile phases in electroosmotic thin-layer chromatography.

    Science.gov (United States)

    Berezkin, V G; Balushkin, A O; Tyaglov, B V; Litvin, E F

    2005-08-19

    A variant of electroosmotic thin-layer chromatography is suggested with the use of low volatility compounds as mobile phases aimed at drastically decreasing the evaporation of the mobile phase and improving the reproducibility of the method. The linear movement velocity of zones of separated compounds is experimentally shown to increase 2-12-fold in electroosmotic chromatography (compared to similar values in traditional TLC). The separation efficiency is also considerably increased.

  1. The weight of flash chromatography: A tool to predict its mass intensity from thin-layer chromatography

    Directory of Open Access Journals (Sweden)

    Freddy Pessel

    2016-11-01

    Full Text Available Purification by flash chromatography strongly impacts the greenness of a process. Unfortunately, due to the lack of the relevant literature data, very often this impact cannot be assessed thus preventing the comparison of the environmental factors affecting the syntheses. We developed a simple mathematical approach to evaluate the minimum mass intensity of flash chromatography from the retention factor values determined by thin-layer chromatography.

  2. Fractionation of fecal neutral steroids by high performance liquid chromatography

    International Nuclear Information System (INIS)

    Jackson, E.M.; Kloss, C.A.; Weintraub, S.T.; Mott, G.E.

    1985-01-01

    Fecal neutral steroids were fractionated by high performance liquid chromatography (HPLC) into three major fractions: 5 beta-H, 3-keto steroids; 5 beta-H, 3 beta-hydroxy steroids; and 5 alpha-H and delta 5-3 beta-hydroxy steroids. This separation was achieved in about 10 minutes, with greater than 97% recovery of standards in each fraction. Gas-liquid chromatographic quantitation of fecal steroids fractionated by either HPLC or thin-layer chromatography gave nearly identical results. A method using both C18 reverse phase and silica HPLC to purify radiolabeled sterols is also described

  3. Thin-layer chromatography and colorimetric analysis of multi-component explosive mixtures

    Science.gov (United States)

    Pagoria, Philip F.; Mitchell, Alexander R.; Whipple, Richard E.; Carman, M. Leslie

    2014-08-26

    A thin-layer chromatography method for detection and identification of common military and peroxide explosives in samples includes the steps of provide a reverse-phase thin-layer chromatography plate; prepare the plate by marking spots on which to deposit the samples by touching the plate with a marker; spot one micro liter of a first standard onto one of the spots, spot one micro liter of a second standard onto another of the spots, and spot samples onto other of spots producing a spotted plate; add eluent to a developing chamber; add the spotted plate to the developing chamber; remove the spotted plate from the developing chamber producing a developed plate; place the developed plate in an ultraviolet light box; add a visualization agent to a dip tank; dip the developed plate in the dip tank and remove the developed plate quickly; and detect explosives by viewing said developed plate.

  4. A reversed-phase compatible thin-layer chromatography autography for the detection of acetylcholinesterase inhibitors.

    Science.gov (United States)

    Ramallo, I Ayelen; García, Paula; Furlan, Ricardo L E

    2015-11-01

    A dual readout autographic assay to detect acetylcholinesterase inhibitors present in complex matrices adsorbed on reversed-phase or normal-phase thin-layer chromatography plates is described. Enzyme gel entrapment with an amphiphilic copolymer was used for assay development. The effects of substrate and enzyme concentrations, pH, incubation time, and incubation temperature on the sensitivity and the detection limit of the assay were evaluated. Experimental design and response surface methodology were used to optimize conditions with a minimum number of experiments. The assay allowed the detection of 0.01% w/w of physostigmine in both a spiked Sonchus oleraceus L. extract chromatographed on normal phase and a spiked Pimenta racemosa (Mill.) J.W. Moore leaf essential oil chromatographed on reversed phase. Finally, the reversed-phase thin-layer chromatography assay was applied to reveal the presence of an inhibitor in the Cymbopogon citratus (DC.) Stapf essential oil. The developed assay is able to detect acetylcholinesterase inhibitors present in complex matrixes that were chromatographed in normal phase or reversed-phase thin-layer chromatography. The detection limit for physostigmine on both normal and reversed phase was of 1×10(-4) μg. The results can be read by a change in color and/or a change in fluorescence. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. High-Performance Thin-Layer Chromatographic Quantification of Rosmarinic Acid and Rutin in Abnormal Savda Munziq

    Directory of Open Access Journals (Sweden)

    S. G. Tian

    2013-01-01

    Full Text Available A high-performance thin-layer chromatographic (HPTLC method has been established for simultaneous analysis of rosmarinic acid and rutin in Abnormal Savda Munziq (ASMq. A methanol extract of ASMq was used for quantification. The compounds were separated on silica gel H thin layer plate with ethyl acetate-formic acid-acetic acid-water 15 : 1 : 1 : 1.5 (v/v as a developer, trichloroethanol as the color reagent. The plates were scanned at 365 nm. The linear calibration data of rosmarinic acid and rutin were in the range of 0.0508 to 0.2540 μg (r=0.9964, 0.2707 to 1.35354 μg (r=0.9981, respectively. The recovery rate of rosmarinic acid was 99.17% (RSD = 2.92% and rutin was 95.24% (RSD = 2.38%. The method enables rapid screening, precise, selective, and sensitive quantification for pharmaceutical analysis.

  6. Reversed-phase thin-layer chromatography of the rare earth elements

    International Nuclear Information System (INIS)

    Kuroda, R.; Adachi, M.; Oguma, K.

    1988-01-01

    Partition chromatographic behaviour of the rare earth elements on C 18 bonded silica reversed-phase material has been investigated by thin-layer chromatography in methanol - lactate media. The rare earth lactato complexes are distributed and fractionated on bonded silica layers without ion-interaction reagents. The concentration and pH of lactate solution, methanol concentration and temperature have effects on the migration and resolution of the rare earth elements. The partition system is particularly suited to separate adjacent rare earths of middle atomic weight groups, allowing the separation of gadolinium, terbium, dysprosium, holmium, erbium and thulium to be achieved by development to 18 cm distance. (orig.)

  7. Extraction of oxytetracycline starting from the meats of chickens and identifications by thin layer chromatography

    International Nuclear Information System (INIS)

    Elghozzi, Amira

    2007-01-01

    Use of antibiotics in the poultry also suspected as one of the cause of the emergence of resistance to antibiotics of some bacteria what causes risks on human health continuation of persistence of the residues in the foodstuffs. We were interested in detected the presence qualitatively of oxytetracyclin in samples of muscle and liver of chicken by the use of the techniques of thin layer chromatography. Although, These results are satisfactory, it is always necessary to prevent the dangers which cause the risks of the residues of oxytetracycline on health. (Author). 20 refs

  8. HIGH PERFORMANCE THIN LAYER CHROMATOGRAPHIC DETERMINATION OF CAFFEIC ACID AND ROSMARINIC ACID FROM THE LEAVES OF Orthosiphon stamineus

    Directory of Open Access Journals (Sweden)

    M. Amzad Hossain

    2010-06-01

    Full Text Available This paper presents the studies performed on extraction of Orthosiphon stamineus, Benth by using different solvent for the identification and quantification of the caffeic acid derivatives such as caffeic acid  and rosmarinic acid which confers to the leaves of this plant with remarkable pharmaceutical properties. High performance thin-layer chromatographic (HPTLC allows the identification and the quantification of more than 20 samples in the same chromatographic run. The analysis of the samples requires 15-30 min compared with more than 2 h using a typical HPLC method. Using the techniques of the HPTLC and the UV-VIS spectra we have found that the extraction of this herb plant contain, the caffeic acid and rosmarinic acid ranging between 0.029% up to 0.506% and up to 0.24% to 2.24% respectively.     Keywords: Caffice acid derivatives, quantification, Malaysian Orthosiphon stamineus, HPTLC

  9. [Analysis of pigments from Rhodotorula glutinis by Raman spectroscopy and thin layer chromatography].

    Science.gov (United States)

    Yuan, Yu-feng; Tao, Zhan-hua; Wang, Xue; Li, Yong-qing; Liu, Jun-xian

    2012-03-01

    The pigments from Rhodotorula glutinis were separated by using thin layer chromatography, and the result showed that Rhodotorula glutinis cells could synthesize at least three kinds of pigments, which were beta-carotene, torulene, and torularhodin. The Raman spectra based on the three pigments were acquired, and original spectra were preprocessed by background elimination, baseline correction, and three-point-smoothing, then the averaged spectra from different pigments were investigated, and the result indicated that Raman shift which represents C-C bond was different, and the wave number of beta-carotene demonstrated the largest deviation, finally torulene and torularhodin in Rhodotorula glutinis had more content than beta-carotene. Quantitative analysis of Raman peak height ratio revealed that peak height ratio of pigments showed little difference, which could be used as parameters for further research on living cells, providing reference content of pigments. The above results suggest that Raman spectroscopy combined with thin layer chromatography can be applied to analyze pigments from Rhodotorula glutinis, provides abundant information about pigments, and serves as an effective method to study pigments.

  10. Stability indicating high performance thin-layer chromatographic method for simultaneous estimation of pantoprazole sodium and itopride hydrochloride in combined dosage form

    Directory of Open Access Journals (Sweden)

    Deepak Bageshwar

    2011-11-01

    Full Text Available A specific, precise and stability indicating high-performance thin-layer chromatographic method for simultaneous estimation of pantoprazole sodium and itopride hydrochloride in pharmaceutical formulations was developed and validated. The method employed TLC aluminium plates precoated with silica gel 60F254 as the stationary phase. The solvent system consisted of methanol:water:ammonium acetate; 4.0:1.0:0.5 (v/v/v. This system was found to give compact and dense spots for both itopride hydrochloride (Rf value of 0.55±0.02 and pantoprazole sodium (Rf value of 0.85±0.04. Densitometric analysis of both drugs was carried out in the reflectance–absorbance mode at 289 nm. The linear regression analysis data for the calibration plots showed a good linear relationship with R2=0.9988±0.0012 in the concentration range of 100–400 ng for pantoprazole sodium. Also, the linear regression analysis data for the calibration plots showed a good linear relationship with R2=0.9990±0.0008 in the concentration range of 200–1200 ng for itopride hydrochloride. The method was validated for specificity, precision, robustness and recovery. Statistical analysis proves that the method is repeatable and selective for the estimation of both the said drugs. As the method could effectively separate the drug from its degradation products, it can be employed as a stability indicating method. Keywords: Gastroesophageal reflux disease (GERD, Itopride hydrochloride, Pantoprazole sodium, High performance thin layer chromatography (HPTLC, Stability indicating, Forced degradation

  11. Determination of radiochemical yields of 186Re-labelled complexes using thin layer chromatography

    International Nuclear Information System (INIS)

    Konirova, R.; Kohlickova, M.; Jedinakova-Krizova, V.

    1999-01-01

    The reaction conditions for synthesis of three rhenium complexes 186 Re-methylendiphosphonate (MDP), 186 Re-hydroxyethylidendiphosphonate (HEDP) and 186 Re-citrate have been investigated. Radiochemical yield of complexation has been determined by thin layer chromatography and paper chromatography. The rhenium complexation with corresponding ligand is dependent on pH values of reaction mixture, concentration of studied ligand (MDP, HEDP and sodium citrate) and concentration of reducing agent. Stannous chloride with ascorbic acid (as antioxidant) was used for reduction of perrhenate. The labeling yield of 186 Re-MDP was about 90 %, of 186 Re-HEDP more than 80 % and more than 75 % for 186 Re-citrate under optimum conditions. Besides, the possibility of application of porphyrins as organic ligands for complexation with rhenium isotopes is examined. (authors)

  12. Reversed-phase thin-layer chromatography behavior of aldopentose derivatives

    Directory of Open Access Journals (Sweden)

    Malbaša Radomir V.

    2012-01-01

    Full Text Available Quantitative structure-retention relationships (QSRR have been used to study the chromatographic behavior of some aldopentose. The behavior of aldopentose derivatives was investigated by means of the reversed-phase thin-layer chromatography (RP TLC on the silica gel impregnated with paraffin oil stationary phases. Binary mixtures of methanol-water, acetone-water and dioxane-water were used as mobile phases. Retention factors, RM0, corresponding to zero percent organic modifier in the aqueous mobile phase was determined. Lipophilicity C0 was calculated as the ratio of the intercept and slope values. There was satisfactory correlation between them and log P values calculated using different theoretical procedures. Some of these correlations offer very good predicting models, which are important for a better understanding of the relationships between chemical structure and retention. The study showed that the hydrophobic parameters RM0 and C0 can be used as a measures of lipophilicity of investigated compounds.

  13. DIFFERENTIATION OF Curcuma longa, Curcuma xanthorrhiza and Zingiber cassumunar BY THIN LAYER CHROMATOGRAPHY FINGERPRINT ANALYSIS

    Directory of Open Access Journals (Sweden)

    Mohamad Rafi

    2011-07-01

    Full Text Available Turmeric (Curcuma longa, java turmeric (Curcuma xanthorrhiza and cassumunar ginger (Zingiber cassumunar are widely used in traditional Indonesian medicine. These three herbs have relatively similar rhizomes colour so it is difficult to be differentiated especially if they are in powder form. A rapid and reliable method, thin layer chromatography (TLC fingerprint, has been developed in order to identify, authenticate and differentiate these three herbs through fingerprint profile of chemical compounds. TLC fingerprints of the three herbs were obtained by visualization of separate zones with visible and UV (254 and 366 nm light. The TLC fingerprint pattern is different each other and showed a specific marker zones respectively. Therefore, TLC fingerprint can be utilized for identification, authentication and differentiation method in quality control of the three herbs tested.

  14. Digitally enhanced thin layer chromatography: further development and some applications in isotopic chemistry.

    Science.gov (United States)

    Manthorpe, Daniel P; Lockley, William J S

    2013-09-01

    Improvements to thin layer chromatography (TLC) analysis can be made easily and cheaply by the application of digital colour photography and image analysis. The combined technique, digitally enhanced TLC (DE-TLC), is applicable to the accurate quantification of analytes in mixtures, to reaction monitoring and to other typical uses of TLC. Examples are given of the application of digitally enhanced TLC to: the deuteromethylations of theophylline to [methyl-(2)H3]caffeine and of umbelliferone to [(2)H3]7-methoxycoumarin; the selection of tertiary amine bases in deuterodechlorination reactions; stoichiometry optimisation in the borodeuteride reduction of quinizarin (1,4-dihydroxyanthraquinone) and to the assessment of xanthophyll yields in Lepidium sativum seedlings grown in deuterated media. Copyright © 2013 John Wiley & Sons, Ltd.

  15. Stability indicating high performance thin layer chromatographic method for quantitation of venlafaxine in bulk and pharmaceutical dosage form

    Directory of Open Access Journals (Sweden)

    Sunil K Dubey

    2015-01-01

    Full Text Available Background: Venlafaxine (VEN is a phenethylamine bicyclic compound, chemically, 1-(2-[dimethyl amino]-1-[4-methoxy phenyl] ethyl cyclo-hexan-1ol hydrochloride. It is a antidepressant. It inhibits the reuptake of serotonin, nor adrenaline and dopamine to a lesser extent at the presynaptic membrane. Aim: A simple, rapid, precise, accurate, and economical high performance thin layer chromatographic (HPTLC method has been developed and validated for the determination of VEN both as a bulk drug and in formulation. Materials and Methods: The method uses aluminum plates precoated with silica gel 60 F254 as the stationary phase and dichloromethane:acetonitrile:N-hexane:triethylamine: 0.5:0.5:4:0.7 (v/v/v/v as mobile phase. Results: This system gave compact spots for VEN (R f = 0.46 ± 0.05. Forced degradation studies were done by subjecting VEN to acid and alkali hydrolysis, oxidation, and reduction. The peak of the degradation product was well resolved from that of the pure drug and had significant different R f values. Analysis of VEN was performed in the absorbance mode at 225 nm. The limit of detection and quantification were 12.48 and 37.81 ng/spot respectively. Conclusions: The developed method was validated and found to be simple, specific, accurate and precise and can be used for routine quality control analysis of VEN in bulk and pharmaceutical formulation.

  16. DETECTION, ISOLATION, AND IDENTIFICATION OF TRUXILLINES IN ILLICIT COCAINE BY MEANS OF THIN-LAYER CHROMATOGRAPHY AND MASS-SPECTROMETRY

    NARCIS (Netherlands)

    ENSING, JG; DEZEEUW, RA

    By means of thin-layer chromatography, an unidentified alkaloidal fraction was observed in illicit cocaine. Because of its persisting presence, efforts were undertaken to isolate and identify this fraction. Various analytical techniques showed complex results, finally pointing to the possibility of

  17. Extraction of Nutraceuticals from Spirulina (Blue-Green Alga): A Bioorganic Chemistry Practice Using Thin-layer Chromatography

    Science.gov (United States)

    Herrera Bravo de Laguna, Irma; Toledo Marante, Francisco J.; Luna-Freire, Kristerson R.; Mioso, Roberto

    2015-01-01

    Spirulina is a blue-green alga (cyanobacteria) with high nutritive value. This work provides an innovative and original approach to the consideration of a bioorganic chemistry practice, using Spirulina for the separation of phytochemicals with nutraceutical characteristics via thin-layer chromatography (TLC) plates. The aim is to bring together…

  18. Thin layer chromatography: a simple and reliable technique for the determination of pesticides residue in environmental samples

    International Nuclear Information System (INIS)

    Asi, M.R.; Hussain, A.; Iqbal, Z.; Chaudhary, J.A.

    2000-01-01

    A method for the detection of pesticides by thin-layer chromatography is described. These pesticides on chlorination and treatment with 0-toluidine, yield an intensively colored blue product. Some of the organophosphates and carbonates can be determined at low level (mu g/ g) by this method. The limit of minimum detectable quantity is 10-100 ng. (author)

  19. Gradient High Performance Liquid Chromatography Method ...

    African Journals Online (AJOL)

    Purpose: To develop a gradient high performance liquid chromatography (HPLC) method for the simultaneous determination of phenylephrine (PHE) and ibuprofen (IBU) in solid ..... nimesulide, phenylephrine. Hydrochloride, chlorpheniramine maleate and caffeine anhydrous in pharmaceutical dosage form. Acta Pol.

  20. The laser desorption/laser ionization mass spectra of some methylated xanthines and the laser desorption of caffeine and theophylline from thin layer chromatography plates

    Science.gov (United States)

    Rogers, Kevin; Milnes, John; Gormally, John

    1993-02-01

    Laser desorption/laser ionization time-of-flight mass spectra of caffeine, theophylline, theobromine and xanthine are reported. These mass spectra are compared with published spectra obtained using electron impact ionization. Mass spectra of caffeine and theophylline obtained by IR laser desorption from thin layer chromatography plates are also described. The laser desorption of materials from thin layer chromatography plates is discussed.

  1. Bacterial Cell Wall Precursor Phosphatase Assays Using Thin-layer Chromatography (TLC) and High Pressure Liquid Chromatography (HPLC).

    Science.gov (United States)

    Pazos, Manuel; Otten, Christian; Vollmer, Waldemar

    2018-03-20

    Peptidoglycan encases the bacterial cytoplasmic membrane to protect the cell from lysis due to the turgor. The final steps of peptidoglycan synthesis require a membrane-anchored substrate called lipid II, in which the peptidoglycan subunit is linked to the carrier lipid undecaprenol via a pyrophosphate moiety. Lipid II is the target of glycopeptide antibiotics and several antimicrobial peptides, and is degraded by 'attacking' enzymes involved in bacterial competition to induce lysis. Here we describe two protocols using thin-layer chromatography (TLC) and high pressure liquid chromatography (HPLC), respectively, to assay the digestion of lipid II by phosphatases such as Colicin M or the LXG toxin protein TelC from Streptococcus intermedius . The TLC method can also monitor the digestion of undecaprenyl (pyro)phosphate, whereas the HPLC method allows to separate the di-, mono- or unphosphorylated disaccharide pentapeptide products of lipid II.

  2. Thin-layer chromatography of polyphosphoinositides from platelet extracts: interference by an unknown phospholipid

    International Nuclear Information System (INIS)

    Tysnes, O.B.; Aarbakke, G.M.; Verhoeven, A.J.; Holmsen, H.

    1985-01-01

    Different ratios of radioactive polyphosphoinositides in platelets pulse-labelled with 32p-orthophosphate have been reported by various laboratories. We studied whether these differences originate from differences in methodology. Extracts of 32p-Pi labelled human platelets were prepared at various times after gel-filtration and phosphatidylinositol (PI)-, mono (PIP)- and bisphosphate (PIP2) were separated by thin-layer chromatography using four different solvent systems. The 32p-levels in PIP and PIP2 remained constant during one hour after gel-filtration, whereas 32p-PI increased continuously and more than doubled within the first h. In two of the systems PIP co-chromatographed with a radioactive compound which separated well from PIP in the two other systems. This unknown compound was also labelled with 3H-glycerol, 3H-inositol and 3H-arachidonic acid, but it was metabolically and functionally different from the polyphosphoinositides. Both the co-chromatography of this unknown phospholipid and the increase in 32p-PI in gel-filtered platelets can explain the difference in 32p-labelling in phosphoinositides reported in the literature

  3. The reciprocal iso-inhibition volume concept: A procedure for the evaluation in effect-directed analysis with thin-layer chromatography - using the thin-layer chromatography-luminescent bacteria assay as an example.

    Science.gov (United States)

    Schulz, Wolfgang; Weiss, Stefan C; Weber, Walter H; Winzenbacher, Rudi

    2017-10-13

    In effect-directed analysis (EDA) with high-performance thin-layer chromatography (HPTLC), the effect is often detected using images. Thus, an approach to create inhibition chromatograms from these images was developed using the example of the HPTLC- bioluminescence inhibition test. A comparison between the cuvette test and the HPTLC test shows that the test on the plate is significantly more sensitive. To describe the strength of the effect, the EC 50 value is determined from the dose-response relationship. However, the inhibiting compounds are generally unknown and thus their concentrations are also unknown. Therefore, instead of the concentration, the known application volumes are used. This enables the calculation of the application volume necessary to achieve 50% inhibition. Since the volume is inversely proportional to the concentration, the reciprocal value of the calculated volume is indicated and is referred to as the reciprocal iso-inhibition volume (RIV). Using this RIV-concept, it is now possible to compare inhibition bands within and between plates. The entire evaluation is described by the means of two samples from a contaminated site using the bioluminescence inhibition. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Analysis of low active-pharmaceutical-ingredient signal drugs based on thin layer chromatography and surface-enhanced Raman spectroscopy.

    Science.gov (United States)

    Li, Xiao; Chen, Hui; Zhu, Qingxia; Liu, Yan; Lu, Feng

    2016-11-30

    Active pharmaceutical ingredients (API) embedded in the excipients of the formula can usually be unravelled by normal Raman spectroscopy (NRS). However, more and more drugs with low API content and/or low Raman scattering coefficient were insensitive to NRS analysis, which was for the first time defined as Low API-Signal Drugs (LASIDs) in this paper. The NRS spectra of these LASIDs were similar to their dominant excipients' profiles, such as lactose, starch, microcrystalline cellulose (MCC), etc., and were classified into three types as such. 21 out of 100 kinds of drugs were screened as LASIDs and characterized further by Raman microscopic mapping. Accordingly, we proposed a tailored solution to the qualitation and quantitation problem of these LASIDs, using surface-enhanced Raman spectroscopic (SERS) detection on the thin layer chromatographic (TLC) plate both in situ and after-separation. Experimental conditions and parameters including TLC support matrix, SERS substrate, detection mode, similarity threshold, internal standard, etc., were optimized. All LASIDs were satisfactorily identified and the quantitation results agreed well with those of high performance liquid chromatography (HPLC). For some structural analogues of LASIDs, although they presented highly similar SERS spectra and were tough to distinguish even with Raman microscopic mapping, they could be successfully discriminated from each other by coupling SERS (with portable Raman spectrometer) with TLC. These results demonstrated that the proposed solution could be employed to detect the LASIDs with high accuracy and cost-effectiveness. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Utilization of thin-layer chromatography for confirmational sampling during remedial excavation

    International Nuclear Information System (INIS)

    Burton, P.J.

    1996-01-01

    Thin-layer chromatography (TLC) is a simple, efficient, inexpensive, and accurate method of chemical analysis. This method is applicable to field testing for confirming the limits of excavation during excavation of petroleum, oil, and/or lubricant (POL) contamination. Traditionally, excavation at contaminated sites suffers from a time delay due to the need to obtain laboratory confirmation of the limits of excavation. All suspected contaminated material is removed for stockpiling or remediation. The remaining soils need to be sampled to confirm that no contaminated material remains in the excavation. The site scientist collects samples and sends them out for analysis at a certified laboratory. The laboratory requires, even for a rush order, several days to analyze the samples. This time delay interferes with the efficiency of the operation. The excavations either have to be left open (which is a safety hazard) or reopened if additional material must be excavated. TLC samples can be collected and analyzed in several hours allowing for a short turnaround time for analytical results. The TLC method can be easily performed by a technician. Results of a case study from a subarctic Alaskan site will be reported. Simple operational TLC procedures will be shared. The equipment required for TLC analysis will be outlined. Correlation data between TLC and laboratory analysis will be presented

  6. Development and validation of a thin-layer chromatography method for stability studies of naproxen

    International Nuclear Information System (INIS)

    Rodriguez Hernandez, Yaslenis; Suarez Perez, Yania; Garcia Pulpeiro, Oscar; Rodriguez Borges, Tania

    2011-01-01

    The validation of an analytical method was carried out to be applied to the stability studies of the future formulations of naproxen suppositories for infant and adult use. The factors which mostly influenced in the naproxen stability were determined, the major degradation occurred in oxidizing acid medium and by action of light. The possible formation of esters between the free carboxyl group present in naproxen and the glyceryl monoestereate present in the base was identified as one of the degradation paths in the new formulation. The results were satisfactory. A thin-layer chromatography-based method was developed as well as the best chromatographic conditions were selected. GF 254 silica gel plates and ultraviolet developer at 254 nm were employed. Three solvent systems were evaluated of which A made up of glacial acetic: tetrahydrofurane:toluene (3:9:90 v/v/v)allowed adequate resolution between the analyte and the possible degradation products, with detection limit of 1 μg. The use of the suggested method was restricted to the identification of possible degradation products just for qualitative purposes and not as final test. The method proved to be sensitive and selective enough to be applied for the stated objective, according to the validation results

  7. Determination of plasma aldosterone in children by thin layer chromatography and radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Parth, K; Zimprich, H; Brunel, R [The Ludwig Boltzmann Institute of Paediatric Endocrinology; The Karolinen Kinderspital, Vienna)

    1976-01-01

    An accurate and relatively simple radioimmunoassay for the determination of aldosterone concentration in peripheral plasma has been developed. 0.5-2.0 ml plasma with added ..cap alpha..1.2-/sup 3/H..omega..aldosterone is extracted with dichloromethane. Purification of the extract is achieved by thin layer chromatography in the system benzene-acetone 1:1. Recovery of ..cap alpha..1,2-/sup 3/H..omega..aldosterone is 58+-6 (sd) %. Bound and free fractions are separated by dextran-coated charcol. The intra-assay reproducibility is 8.8 % and the inter-assay reproducibility varies from 11.4-16.1%. The sensitivity of the assay for a 5 ml plasma sample can be put at 0.2 ng/100 ml. Normal values determined in 52 healthy children of different age groups are presented. Furthermore the aldosterone stimulating effect of low sodium diet (17 children), severe and prolonged vomiting (19 children) and synthetic ACTH (10 children) has been studied by our modified method.

  8. Molecular Ionization-Desorption Analysis Source (MIDAS) for Mass Spectrometry: Thin-Layer Chromatography

    Science.gov (United States)

    Winter, Gregory T.; Wilhide, Joshua A.; LaCourse, William R.

    2016-02-01

    Molecular ionization-desorption analysis source (MIDAS), which is a desorption atmospheric pressure chemical ionization (DAPCI) type source, for mass spectrometry has been developed as a multi-functional platform for the direct sampling of surfaces. In this article, its utility for the analysis of thin-layer chromatography (TLC) plates is highlighted. Amino acids, which are difficult to visualize without staining reagents or charring, were detected and identified directly from a TLC plate. To demonstrate the full potential of MIDAS, all active ingredients from an analgesic tablet, separated on a TLC plate, were successfully detected using both positive and negative ion modes. The identity of each of the compounds was confirmed from their mass spectra and compared against standards. Post separation, the chemical signal (blue permanent marker) as reference marks placed at the origin and solvent front were used to calculate retention factor (Rf) values from the resulting ion chromatogram. The quantitative capabilities of the device were exhibited by scanning caffeine spots on a TLC plate of increasing sample amount. A linear curve based on peak are, R2 = 0.994, was generated for seven spots ranging from 50 to 1000 ng of caffeine per spot.

  9. Surface enhanced Raman scattering imaging of developed thin-layer chromatography plates.

    Science.gov (United States)

    Freye, Chris E; Crane, Nichole A; Kirchner, Teresa B; Sepaniak, Michael J

    2013-04-16

    A method for hyphenating surface enhanced Raman scattering (SERS) and thin-layer chromatography (TLC) is presented that employs silver-polymer nanocomposites as an interface. Through the process of conformal blotting, analytes are transferred from TLC plates to nanocomposite films before being imaged via SERS. A procedure leading to maximum blotting efficiency was established by investigating various parameters such as time, pressure, and type and amount of blotting solvent. Additionally, limits of detection were established for test analytes malachite green isothiocyanate, 4-aminothiophenol, and Rhodamine 6G (Rh6G) ranging from 10(-7) to 10(-6) M. Band broadening due to blotting was minimal (∼10%) as examined by comparing the spatial extent of TLC-spotted Rh6G via fluorescence and then the SERS-based spot size on the nanocomposite after the blotting process. Finally, a separation of the test analytes was carried out on a TLC plate followed by blotting and the acquisition of distance × wavenumber × intensity three-dimensional TLC-SERS plots.

  10. On-chip ultra-thin layer chromatography and surface enhanced Raman spectroscopy.

    Science.gov (United States)

    Chen, Jing; Abell, Justin; Huang, Yao-wen; Zhao, Yiping

    2012-09-07

    We demonstrate that silver nanorod (AgNR) array substrates can be used for on-chip separation and detection of chemical mixtures by combining ultra-thin layer chromatography (UTLC) and surface enhanced Raman spectroscopy (SERS). The UTLC-SERS plate consists of an AgNR array fabricated by oblique angle deposition. The capability of the AgNR substrates to separate the different compounds in a mixture was explored using a mixture of four dyes and a mixture of melamine and Rhodamine 6G at varied concentrations with different mobile phase solvents. After UTLC separation, spatially-resolved SERS spectra were collected along the mobile phase development direction and the intensities of specific SERS peaks from each component were used to generate chromatograms. The AgNR substrates demonstrate the potential for separating the test dyes with plate heights as low as 9.6 μm. The limits of detection are between 10(-5)-10(-6) M. Furthermore, we show that the coupling of UTLC with SERS improves the SERS detection specificity, as small amounts of target analytes can be separated from the interfering background components.

  11. [Analysis of phthalates in plastic food-packaging bags by thin layer chromatography].

    Science.gov (United States)

    Chen, Hui; Wang, Yuan; Zhu, Ruohua

    2006-01-01

    The method for simultaneous determination of four phthalates, namely dimethyl phthalate (DMP), diethyl phthalate (DEP), di-n-butyl phthalate (DBP) and di (2-ethylhexyl) phthalate (DEHP) in plastic food-packaging bags by thin layer chromatography (TLC) was developed. The plastic food-packaging bags were extracted with ethanol by ultrasonication, then the mixture was filtrated through membrane (0.45 microm). The mixture of ethyl acetate-anhydrous ether-isooctane (1 : 4 : 15, v/v) was used as developing agent on the TLC silica gel plate for development. The filtered liquid was spotted on the TLC plate dealt by acetone, and detected with scanning wavelength of 275 nm and reference wavelength of 340 nm. The qualitative analysis of the phthalates was performed using the R(f) values of the chromatogram. The quantitative analysis was performed with external standard method. Good linearities were obtained for DMP, DEP, DBP and DEHP. The detection limits were 2.1 ng for DMP, 2.4 ng for DEP, 3.4 ng for DBP and 4.0 ng for DEHP. The relative standard deviations (RSDs) of the four phthalates were 2.8% - 3.5%. The recoveries of the four phthalate standards in real sample were 78.58% - 111.04%. The method presented has the advantages of high precision, high sensitivity, small sample size, and simple pretreatment . The method was used to detect the four phthalates in the food-packaging bags. The contents in real samples were close to the results by gas chromatography.

  12. Immunoaffinity column clean-up and thin layer chromatography for determination of ochratoxin A in green coffee.

    Science.gov (United States)

    Santos, E A; Vargas, E A

    2002-05-01

    An immunoaffinity clean-up-based method for determining ochratoxin A (OTA) in green coffee aiming at one-dimensional thin layer chromatography (TLC) analysis was established. OTA was extracted with a mixture of methanol and aqueous sodium hydrogen carbonate solution, purified through an immunoaffinity column, separated on normal or reversed-phase (RP) TLC plates and detected and quantified by visual and densitometric analysis. The linear equation of the standard calibration curve by densitometric analysis gave R(2) > 0.999 (0.04-84 ng). The mean recovery (R) of OTA from spiked samples (1.8-109 microg kg(-1)) by densitometric and visual analyses were 98.4 and 103.8%, respectively. The relative standard deviations (RSD) for densitometric and visual analysis varied from 1.1 to 24.9% and from 0.0 to 18.8%, respectively. The RSD for naturally contaminated samples by densitometry (three levels of contamination, n = 3) varied from 11.1 to 18.1%. The correlation (R(2)) between high-performance liquid chromatography (HPLC) and densitometry, and between visual and densitometric analysis for spiked samples were > 0.99. The limit of detection (LOD) of the method was 0.5 microg kg(-1) for normal TLC. Toluene-ethyl acetate-88% formic acid (6:3:1 v/v/v) and acetonitrile-methanol-water-glacial acetic acid (35:35:29:10 v/v/v/v) were regarded as the suitable TLC solvents for eluting both standards and samples on normal and RP TLC plates, respectively. Toluene-acetic acid (99:1 v/v) was chosen as the spotting solvent among several others for giving the best sensitivity and resolution of OTA on TLC plates as well as the best recovery of OTA from standard and sample extract residues. Preliminary studies were carried out to investigate the reuse of the immunoaffinity column and the interference of caffeine in the OTA recovery.

  13. Validated High Performance Liquid Chromatography Method for ...

    African Journals Online (AJOL)

    Purpose: To develop a simple, rapid and sensitive high performance liquid chromatography (HPLC) method for the determination of cefadroxil monohydrate in human plasma. Methods: Schimadzu HPLC with LC solution software was used with Waters Spherisorb, C18 (5 μm, 150mm × 4.5mm) column. The mobile phase ...

  14. The phytochemical screening and thin layer chromatography results of Jatropha gossypiifolia seeds

    Directory of Open Access Journals (Sweden)

    Anis Nurwidayati

    2013-05-01

    , the intermediary for schistosomiasis is widespread in this region. Eradication has been done by spraying chemical molluscicides. This study aimed to identify the class of chemical compounds in the methanol extract of red castor seed.Methods: The study was conducted in May 2009. Red castor seeds were collected from Palu, Central Sulawesi. Red castor seeds extraction was done by percolation method using methanol solvent. Phytochemical screening test was performed with a tube to detect the compound in red castor bean extract. Screening was followed by thin layer chromatography testing to ensure the screening results of the test tube.Results:Extracts that was produced from 500 grams of red castor dry seed powder with 2500 ml of methanol solvent was 250 ml thick reddish brown fluid. Phytochemical screening with a test tube showed positive results of alkaloid by the formation of deposits in Meyer test, Wagner test, and Dragendorff test. Screening the methanol extracts of red castor seed also showed positive results on saponins by foam test and LiebermanBurchard test. Positive results on Killiani Keller tests and Kedde test suggests that red castor bean extract contains cardenoline and bufadienol. Thin-layer chromatography analysis showed that the red castor bean extract is positive for terpenes with the formation of spots on the silica gel plate when terpenes was sprayed (cerium sulfate reagent. Conclusion: Chemical components contained in the methanol extract of red castor beans consisted of alkaloids, saponins, cardenolin, bufadienol, and terpenes. (Health Science Indones 2012;2:xx-xxKeywords: schistosomiasis, Jatropha gossypifolia, chemical compound

  15. Investigation of Symphytum cordatum alkaloids by liquid-liquid partitioning, thin-layer chromatography and liquid chromatography-ion-trap mass spectrometry

    International Nuclear Information System (INIS)

    Mroczek, Tomasz; Ndjoko-Ioset, Karine; Glowniak, Kazimierz; Mietkiewicz-Capala, Agnieszka; Hostettmann, Kurt

    2006-01-01

    From the alkalised crude extract of Symphytum cordatum (L.) W.K. roots, pyrrolizidine alkaloids (PAs) were extracted as free tertiary bases and polar N-oxides in a merely one-step liquid-liquid partitioning (LLP) in separation funnel and subsequently pre-fractionated by preparative multiple-development (MD) thin-layer chromatography (TLC) on silica gel plates. In this way three alkaloid fractions of different polarities and retention on silica gel plates were obtained as: the most polar N-oxides of the highest retention, the tertiary bases of medium retention, and diesterified N-oxides of the lowest retention. The former fraction was reduced into free bases by sodium hydrosulfite and purified by LLP on Extrelut-NT3 cartridge. It was further analysed together with the two other fractions by high-performance liquid chromatography (HPLC)-ion-trap mass spectrometry with atmospheric pressure chemical ionization (APCI) interface on XTerra C 18 column using a gradient elution. Based on MS n spectra, 18 various alkaloids have been tentatively determined for the first time in this plant as the following types of structure: echimidine-N-oxide (three diasteroisomers), 7-sarracinyl-9-viridiflorylretronecine (two diasteroisomers), echimidine (two diasteroisomers), lycopsamine (two diasteroisomers), dihydroechinatine-N-oxide, dihydroheliospathuline-N-oxide, lycopsamine-N-oxide (three diasteroisomers), 7-acetyllycopsamine-N-oxide, symphytine-N-oxide (two diasteroisomers) and 2'',3''-epoxyechiumine-N-oxide

  16. Investigation of Symphytum cordatum alkaloids by liquid-liquid partitioning, thin-layer chromatography and liquid chromatography-ion-trap mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Mroczek, Tomasz [Department of Pharmacognosy with Medicinal Plants Laboratory, Medical University, 1 Chodzki St., 20-093 Lublin (Poland)]. E-mail: tmroczek@pharmacognosy.org; Ndjoko-Ioset, Karine [Laboratoire de Pharmacognosie et Phytochimie, Ecole de Pharmacie Geneve-Lausanne, Universite de Geneve, Quai Ernest-Ansermet 30, CH-1211 Geneva 4 (Switzerland); Glowniak, Kazimierz [Department of Pharmacognosy with Medicinal Plants Laboratory, Medical University, 1 Chodzki St., 20-093 Lublin (Poland); Mietkiewicz-Capala, Agnieszka [Department of Pharmacognosy with Medicinal Plants Laboratory, Medical University, 1 Chodzki St., 20-093 Lublin (Poland); Hostettmann, Kurt [Laboratoire de Pharmacognosie et Phytochimie, Ecole de Pharmacie Geneve-Lausanne, Universite de Geneve, Quai Ernest-Ansermet 30, CH-1211 Geneva 4 (Switzerland)

    2006-05-04

    From the alkalised crude extract of Symphytum cordatum (L.) W.K. roots, pyrrolizidine alkaloids (PAs) were extracted as free tertiary bases and polar N-oxides in a merely one-step liquid-liquid partitioning (LLP) in separation funnel and subsequently pre-fractionated by preparative multiple-development (MD) thin-layer chromatography (TLC) on silica gel plates. In this way three alkaloid fractions of different polarities and retention on silica gel plates were obtained as: the most polar N-oxides of the highest retention, the tertiary bases of medium retention, and diesterified N-oxides of the lowest retention. The former fraction was reduced into free bases by sodium hydrosulfite and purified by LLP on Extrelut-NT3 cartridge. It was further analysed together with the two other fractions by high-performance liquid chromatography (HPLC)-ion-trap mass spectrometry with atmospheric pressure chemical ionization (APCI) interface on XTerra C{sub 18} column using a gradient elution. Based on MS {sup n} spectra, 18 various alkaloids have been tentatively determined for the first time in this plant as the following types of structure: echimidine-N-oxide (three diasteroisomers), 7-sarracinyl-9-viridiflorylretronecine (two diasteroisomers), echimidine (two diasteroisomers), lycopsamine (two diasteroisomers), dihydroechinatine-N-oxide, dihydroheliospathuline-N-oxide, lycopsamine-N-oxide (three diasteroisomers), 7-acetyllycopsamine-N-oxide, symphytine-N-oxide (two diasteroisomers) and 2'',3''-epoxyechiumine-N-oxide.

  17. Purification of 3H-dihydroalprenolol by two dimensional thin layer chromatography

    International Nuclear Information System (INIS)

    Smisterova, J.; Soltes, L.; Kallay, Z.

    1989-01-01

    A two dimensional thin-layer chromatographic method was developed for the purification and analysis of (-)-[ 3 H]dihydroalprenolol by using an acidic mobile phase (butanol/water/acetic acid 25:10:4, v/v) in one direction and a basic eluent (chloroform/acetone/triethylamine 50:40:10, v/v) in another direction. (author)

  18. Application of RF correction in thin-layer chromatography by means of two reference RF values

    NARCIS (Netherlands)

    Dhont, J.H.; Vinkenborg, C.; Compaan, H.; Ritter, F.J.; Labadie, R.P.; Verweij, A.; Zeeuw, R.A. de

    1972-01-01

    Results of the inter-laboratory experiment described in this paper show that the GALANOS AND KAPOULAS equation can be applied satisfactorily to correct RF values obtained on thin-layer chromatograms in a polar multi-component solvent. Addition of Kieselguhr to the silica gel gives RFc values

  19. Validated high-performance thin-layer chromatographic (HPTLC method for simultaneous determination of nadifloxacin, mometasone furoate, and miconazole nitrate cream using fractional factorial design

    Directory of Open Access Journals (Sweden)

    Kalpana G. Patel

    2016-07-01

    Full Text Available A high-performance thin-layer chromatographic method for simultaneous determination of nadifloxacin, mometasone furoate, and miconazole nitrate was developed and validated as per International Conference on Harmonization guidelines. High-performance thin-layer chromatographic separation was performed on aluminum plates precoated with silica gel 60F254 and methanol:ethyl acetate:toluene: acetonitrile:3M ammonium formate in water (1:2.5:6.0:0.3:0.2, % v/v as optimized mobile phase at detection wavelength of 224 nm. The retardation factor (Rf values for nadifloxacin, mometasone furoate, and miconazole nitrate were 0.23, 0.70, and 0.59, respectively. Percent recoveries in terms of accuracy for the marketed formulation were found to be 98.35–99.76%, 99.36–99.65%, and 99.16–100.25% for nadifloxacin, mometasone furoate, and miconazole nitrate, respectively. The pooled percent relative standard deviation for repeatability and intermediate precision studies was found to be < 2% for three target analytes. The effect of four independent variables, methanol content in total mobile phase, wavelength, chamber saturation time, and solvent front, was evaluated by fractional factorial design for robustness testing. Amongst all four factors, volume of methanol in mobile phase appeared to have a possibly significant effect on retention factor of miconazole nitrate compared with the other two drugs nadifloxacin and mometasone furoate, and therefore it was important to be carefully controlled. In summary, a novel, simple, accurate, reproducible, and robust high-performance thin-layer chromatographic method was developed, which would be of use in quality control of these cream formulations.

  20. Thin layer chromatography of glucose and sorbitol on Cu(II)-impregnated silica gel plates

    Energy Technology Data Exchange (ETDEWEB)

    Hadzija, O. (Ruder Boskovic Inst., Zagreb (Croatia)); Spoljar, B. (Ruder Boskovic Inst., Zagreb (Croatia)); Sesartic, L. (Inst. of Immunology, Zagreb (Croatia))

    1994-04-01

    A thin-layer chromatographic (TLC) separation of glucose and sorbitol on CU(II)-impregnated silica gel plates with n-propanol: Water (4:1) v/v as developer and potassium permanganate as detecting reagent has been worked out. The new impregnant is completely insoluble in water and thus enables the use of an aqueous developer. The R[sub f]-values are 55 and 10 for glucose and sorbitol, respectively. (orig.)

  1. In situ Silver Spot Preparation and on-Plate Surface-Enhanced Raman Scattering Detection in Thin Layer Chromatography Separation

    Science.gov (United States)

    Herman, K.; Mircescu, N. E.; Szabo, L.; Leopold, L. F.; Chiş, V.; Leopold, N.

    2013-05-01

    An improved approach for surface-enhanced Raman scattering (SERS) detection of mixture constituents after thin layer chromatography (TLC) separation is presented. A SERS active silver substrate was prepared under open air conditions, directly on the thin silica film by photo-reduction of silver nitrate, allowing the detection of binary mixtures of cresyl violet, bixine, crystal violet, and Cu(II) complex of 4-(2-pyridylazo)resorcinol. The recorded SERS spectrum provides a unique spectral fingerprint for each molecule; therefore the use of analyte standards is avoided, thus rendering the presented procedure advantageous compared to the conventional detection methodology in TLC.

  2. High performance liquid chromatography in pharmaceutical analyses

    Directory of Open Access Journals (Sweden)

    Branko Nikolin

    2004-05-01

    Full Text Available In testing the pre-sale procedure the marketing of drugs and their control in the last ten years, high performance liquid chromatographyreplaced numerous spectroscopic methods and gas chromatography in the quantitative and qualitative analysis. In the first period of HPLC application it was thought that it would become a complementary method of gas chromatography, however, today it has nearly completely replaced gas chromatography in pharmaceutical analysis. The application of the liquid mobile phase with the possibility of transformation of mobilized polarity during chromatography and all other modifications of mobile phase depending upon the characteristics of substance which are being tested, is a great advantage in the process of separation in comparison to other methods. The greater choice of stationary phase is the next factor which enables realization of good separation. The separation line is connected to specific and sensitive detector systems, spectrafluorimeter, diode detector, electrochemical detector as other hyphernated systems HPLC-MS and HPLC-NMR, are the basic elements on which is based such wide and effective application of the HPLC method. The purpose high performance liquid chromatography(HPLC analysis of any drugs is to confirm the identity of a drug and provide quantitative results and also to monitor the progress of the therapy of a disease.1 Measuring presented on the Fig. 1. is chromatogram obtained for the plasma of depressed patients 12 h before oral administration of dexamethasone. It may also be used to further our understanding of the normal and disease process in the human body trough biomedical and therapeutically research during investigation before of the drugs registration. The analyses of drugs and metabolites in biological fluids, particularly plasma, serum or urine is one of the most demanding but one of the most common uses of high performance of liquid chromatography. Blood, plasma or

  3. Determination of Polycyclic Aromatic Hydrocarbons in Automobile Exhaust by Means of High-Performance Liquid Chromatography with Fluorescence Detection

    DEFF Research Database (Denmark)

    Nielsen, Tom

    1979-01-01

    A chromatographic method has been developed and applied to the determination of polycyclic aromatic hydrocarbons (PAHs) in particulate matter in automobile exhaust, in petrols, and in crankcase oils. The PAHs were purified from other organic compounds by thin-layer chromatography, separated by high......-performance liquid chromatography, and measured by means of on-line fluorescence detection. The identities of the PAHs were verified by comparing the emission spectra obtained by a stop-flow technique with those of standard PAHs...

  4. Simple Identification of the Neutral Chlorinated Auxin in Pea by Thin Layer Chromatography

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen

    1980-01-01

    to small volumes and chromatographed in CHCl3 or CCl4 solvent systems separating the chlorinated auxin from indoleacetonitrile and the methyl or ethyl esters of indoleacetic acid. Colour reaction was carried out with some of the Salkowski FeCl3 sprays of which Ehmann's FeCl3/dimethylaminobenzaldehyde......One of the neutral chlorinated auxins of immature pea seeds was readily identified by thin layer procedures simple enough to serve in student's laboratory courses. 4-Chloroindole-3-acetic acid methyl ester was extracted from 50 g of commercial, frozen peas by either water or acetone, concentrated...

  5. Variables affecting resolution of lung phospholipids in one-dimensional thin-layer chromatography.

    Science.gov (United States)

    Krahn, J

    1987-01-01

    Resolution of the confusion in the literature about the separation of lung phospholipids in thin-layer chromatographic systems has awaited a systematic study of the variables that potentially affect this separation. In this study I show that: incorporation of ammonium sulfate into silica gel "GHL" has a dramatic effect on separation of lung phospholipids; this effect is equally dramatic but different in activated and nonactivated gels; when it picks up moisture, ammonium sulfate-activated gel very rapidly loses its ability to resolve lecithin from phosphatidylinositol; in gel containing ammonium sulfate, small amounts of phosphatidylethanolamine are hydrolyzed to lyso-phosphatidylethanolamine.

  6. Development of a high performance liquid chromatography method ...

    African Journals Online (AJOL)

    Development of a high performance liquid chromatography method for simultaneous ... Purpose: To develop and validate a new low-cost high performance liquid chromatography (HPLC) method for ..... Several papers have reported the use of ...

  7. Separation of Berberine Hydrochloride and Tetrahydropalmatine and Their Quantitative Analysis with Thin Layer Chromatography Involved with Ionic Liquids

    Directory of Open Access Journals (Sweden)

    Jing Lu

    2015-01-01

    Full Text Available [BMIM]OH was used in mobile and stationary phase of thin layer chromatography (TLC to analyze berberine hydrochloride and tetrahydropalmatine for the first time. Supported imidazole ionic liquid with hydroxide ion on silica gel (SiO2·Im+·OH− was synthesized through simple procedure and characterized by Fourier transform infrared spectroscopy (FT-IR, elemental analysis, and scanning electron microscope (SEM. Moreover, on the plates prepared by SiO2·Im+·OH−, the contents of the above alkaloids in the Chinese patent medicine (CPM of “Stomacheasy” capsule were successfully determined by TLC scanner. The key conditions and chromatographic behaviors were also investigated in detail. According to similar ways, ionic liquids (ILs also can be used in other planar chromatographies in two modes. This study is expected to be helpful in expanding the application of IL and its bonded silica gel in TLC separation field.

  8. Identification and determination of oxytetracycline, tiamulin, lincomycin, and spectinomycin in veterinary preparations by thin-layer chromatography/densitometry.

    Science.gov (United States)

    Krzek, J; Kwiecień, A; Starek, M; Kierszniewska, A; Rzeszutko, W

    2000-01-01

    A thin-layer chromatographic/densitometric method was developed for the identification and quantitation of oxytetracycline, tiamulin, lincomycin, and spectinomycin in veterinary preparations. Silica gel-coated thin layer chromatography plates and 2 mobile phases were used to separate these constituents. The appropriate compositions of the suitable mobile phases were established: 10% citric acid solution-n-hexane-ethanol (80 + 1 + 1, v/v) and n-butanol-ethanol-chloroform-25% ammonia (4 + 5 + 2 + 5, v/v). Along with Rf values and spot colors, direct UV and visual densitometric measurements were used for identification. Similar measuring ranges were used for quantitative analysis to obtain repeatable and reliable results for the preparations examined. The results of the quantitative analysis are characterized by a small confidence interval and are close to the declared contents of active constituents: oxytetracycline 30.01 +/- 0.38 g at lambda = 350 nm and 30.24 +/- 0.86 g at lambda = 430 nm; tiamulin, 10.19 +/- 0.86 g at lambda = 450 nm; lincomycin, 2.27 +/- 0.08 g at lambda = 278 nm; and spectinomycin, 2.18 +/- 0.07 g at lambda = 421 nm. The recoveries for all antibiotics ranged from 100.01 to 102.54%.

  9. Role of thin-layer chromatography in ascertaining Kashaya Rasa (astringent taste) in medicinal plants on the concept of Samana and Vichitra Pratyayarabdha principles of Ayurveda.

    Science.gov (United States)

    Kolhe, Rasika H; Acharya, Rabinarayan; Shukla, Vinay J

    2014-04-01

    Pharmacodynamics, in Ayurveda has been described in terms of Rasadipanchaka. Rasa, on one side indicates the Bhautika composition of the drug and on the other side predicts the action. Different analytical techniques, pharmaceutical processes are being used in Ayurveda for the purpose of standardization of raw drugs. In this study an attempt has been made to apply chromatographic technique in determination of Kashaya (astringent) Rasa (taste). Two important Kashaya dominant drugs Kulattha (Dolichos biflorus Linn.) and Kanchanara (Bauhinia variegata Linn.), falling under Vichitra and Samana Pratyayarabdha category respectively, were subjected to physicochemical parameters and qualitative tests followed by High-Performance Thin-Layer Chromatography (HPTLC). In light of chromatographic fingerprinting; sample preparation protocol is modified to incorporate taste threshold in correlation. Column chromatography is used for first-level discrimination technique followed by HPTLC. Kashaya Rasa Dominant Zone (KsRDZ) was separated and subjected to TLC fingerprinting. The KsRDZ fraction was designated as Botanical Reference Material (BRM) in further analysis. Ash value, Alcohol and water soluble extract value were more in B variegata as compared to D biflorus. Presence of tannin in both the samples was confirmed through qualitative test. The KsRDZ fraction separated at Rf 0.46 and 0.48 for Kulattha and Kanchanara respectively. The results showed that the planner chromatography technique seems very useful when BRM hypothesis was adjunct to method that explains the categorization according to traditional Rasa domain classification method.

  10. Solvent Front Position Extraction procedure with thin-layer chromatography as a mode of multicomponent sample preparation for quantitative analysis by instrumental technique.

    Science.gov (United States)

    Klimek-Turek, A; Sikora, E; Dzido, T H

    2017-12-29

    A concept of using thin-layer chromatography to multicomponent sample preparation for quantitative determination of solutes followed by instrumental technique is presented. Thin-layer chromatography (TLC) is used to separate chosen substances and their internal standard from other components (matrix) and to form a single spot/zone containing them at the solvent front position. The location of the analytes and internal standard in the solvent front zone allows their easy extraction followed by quantitation by HPLC. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Monitoring of monooctanoyl phosphatidylcholine synthesis by enzymatic acidolysis between soybean phosphatidylcholine and caprylic acid by thin-layer chromatography with a flame ionization detector

    DEFF Research Database (Denmark)

    Vikbjerg, Anders Falk; Mu, Huiling; Xu, Xuebing

    2005-01-01

    Thin-layer chromatography with flame ionization detector (TLC-FID) method was used for monitoring the production of structured phospholipids (ML-type: L-long chain fatty acids; M-medium chain fatty acids) by enzyme-catalyzed acidolysis between soybean phosphatidylcholine (PC) and caprylic acid....... It was found that the structured PC fractionated into 2-3 distinct bands on both plate thin layer chromatography (TLC) and Chromarod TLC. These 3 bands represented PC of LL-type, ML-type and MM-type, respectively. The TLC-FID method was applied in the present study to examine the influence of enzyme dosage...

  12. Use of two-phase aqueous systems based on water-soluble polymers in thin-layer and extraction chromatography for recovery and separtion of actinides

    International Nuclear Information System (INIS)

    Molochnikova, N.P.; Shkinev, V.M.; Myasoedov, B.F.

    1995-01-01

    The feasibility has been demonstrated of using two-phase aqueous systems based on water-soluble polymers, polyethylene glycol and dextran sulfate, in thin-layer and extraction chromatography for recovery and separation of actinides. A convenient method has been proposed for continuous recovery of 239 Np from 243 Am, originating from differences in sorption of tri- and pentavalent actinides from sulfate solutions containing potassium phosphotungstate by silica gel impregnated with polyethylene glycol. New plates for thin-layer chromatography using water-soluble polymers have been developed. These plates were used to study behavior of americium in various oxidation states in thin sorbent layers

  13. Determination of aflatoxins B1 and M1 in animal feeds and liquid milk using thin layer chromatography

    International Nuclear Information System (INIS)

    Njue, W.; Gitu, L.; Kaberia, F.

    1996-01-01

    Animal feed samples were collected from feeding troughs and analysed for levels of aflatoxins B 1 , a toxic and carcinogenic mycotoxin. When aflatoxin B 1 is consumed by dairy cattle some of it is hydroxylated to form aflatoxin M 1 , which can appear in milk. Since aflatoxin M 1 , is also toxic and carcinogenic, it was determined in liquid milk. The determinations were carried out using thin-layer chromatography. Some of the feed samples were found to contain concentrations of aflatoxin B 1 that were above maximum tolerated values in foods and feeds in various countries. Brewers grain and used poultry feed contained 133.4 ppb, while the barley husks had a maximum value of 27.4 ppb. The details of the experimental results and analytical methods used are presented.(author)

  14. Larvicidal Effect of Vinca Fruit Extract (Vinca rosea Against Aedes aegypti Larvae and Secondary Metabolites Profile by Thin Layer Chromatography

    Directory of Open Access Journals (Sweden)

    Rahmawati Ekaputri

    2014-04-01

    Full Text Available Background: Vinca rosea is known contain alkaloids, it was usually used to treat various diseases. Alkaloids from Vinca leaves are also already known have larvicidal activity. Based on this toxicological activity, the fruit of Vinca rosea was selected to investigation its larvicidal activity against the 3rd instar larvae of the mosquito vector of dengue haemorrhagic fever (DHF Aedes aegypti.  Five concentrations of Vinca fruit extract were tested against the 3rd instar Aedes aegypti larvae. The different larval mortality percentages were recorded after 24 hours. Lethal concentration (LC50 anf LC90 of Vinca fruit extract were calculated using Probit analysis. Phytochemical compounds  of ethanolic extract also investigated using Thin layer Chromatography (TLC. LC50 and LC90 values of fruit extract were 2.987 mg/ml and 32.861 mg/ml. Alkaloids were detected in extract.

  15. Profiling the triacylglyceride contents in bat integumentary lipids by preparative thin layer chromatography and MALDI-TOF mass spectrometry.

    Science.gov (United States)

    Pannkuk, Evan L; Risch, Thomas S; Savary, Brett J

    2013-09-05

    The mammalian integument includes sebaceous glands that secrete an oily material onto the skin surface. Sebum production is part of the innate immune system that is protective against pathogenic microbes. Abnormal sebum production and chemical composition are also a clinical symptom of specific skin diseases. Sebum contains a complex mixture of lipids, including triacylglycerides, which is species-specific. The broad chemical properties exhibited by diverse lipid classes hinder the specific determination of sebum composition. Analytical techniques for lipids typically require chemical derivatizations that are labor-intensive and increase sample preparation costs. This paper describes how to extract lipids from mammalian integument, separate broad lipid classes by thin-layer chromatography, and profile the triacylglyceride contents using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. This robust method enables a direct determination of the triacylglyceride profiles among species and individuals, and it can be readily applied to any taxonomic group of mammals.

  16. The characterization of petroleum and creosote-contaminated soils: Class component analysis by thin layer chromatography with flame ionization detection

    International Nuclear Information System (INIS)

    Pollard, S.J.T.; Hrudey, S.E.; Fuhr, B.J.; Alex, R.F.; Holloway, L.R.

    1992-01-01

    The assessment and reclamation of coal tar, creosote and petroleum-contaminated sites is emerging as a major challenge to industry and the federal and provincial Canadian governments. Contaminants frequently include polynuclear aromatic compounds (PAH), several high molecular weight analogues of which are documented carcinogens. Adaptation of thin layer chromatography with flame ionization detection for the analysis of hydrocarbon-contaminated soils is described. The method is directly applicable to the analysis of oily waste extracts from petroleum and creosote wood preservative site soils and is capable of distinguishing between the saturate, aromatic and polar components of waste residues. The method provides a rapid, low cost class component analysis of heavy hydrocarbon waste extracts and is particularly useful for estimating the extent of weathering experienced by chronically exposed hydrocarbon wastes in the soil environment. As such, it is useful as a screening tool for a preliminary assessment of the biotreatability or inherent recalcitrance of hydrocarbon waste mixtures. 11 refs., 5 figs

  17. Separation of metals in the form of ion associates by the method of thin-layer chromatography

    International Nuclear Information System (INIS)

    Shapovalova, E.N.; Timerbaev, A.R.; Bol'shova, T.A.; Mel'nik, S.V.; Kordejro, E.

    1990-01-01

    Behaviour of pyridylazo resorcinates of certain metals (Ga, In, Fe, Co) in the form of ionic pairs with tri-n-octylamine (TOA) under conditions of thin-layer chromatography (TLC) has been studied. For all eluents investigated Ga and In complexes possess the highest mobility. Selectivity of ionic associate separation decreases with an increase in mobile phase polarity. Mixtures with 10-15 % content of isopropanol in eluating solution are the optimal ones. Separation of Ga and In from Fe 3+ and Co takes place with separation criterion 3.1 and 4.1 respectively. An attempt to separate ionic associates of In and Ga failed owing to similar stability of their pyridylazoresorcinates. Solution of the problem of In and Ga determination in the presence of iron can contribute to concrete application of the method

  18. Ultra-thin layer chromatography and surface enhanced Raman spectroscopy on silver nanorod array substrates prepared by oblique angle deposition

    Science.gov (United States)

    Chen, Jing; Abell, Justin; Huang, Yao-wen; Zhao, Yiping

    2012-06-01

    We demonstrate the potential use of silver nanorod (AgNR) array substrates for on-chip separation and detection of chemical mixtures by ultra-thin layer chromatography (UTLC) and surface enhanced Raman spectroscopy (SERS). The capability of the AgNR substrates to separate different compounds in a mixture was explored using a mixture of the food colorant Brilliant Blue FCF and lactic acid, and the mixtures of Methylene Violet and BSA at various concentrations. After the UTLC process, spatially-resolved SERS spectra were collected along the mobile phase development direction and the intensities of specific SERS peaks from each component were used to generate chromatograms. The AgNR substrates demonstrate the capability of separating Brilliant Blue from lactic acid, as well as revealing the SERS signal of Methylene Violet from the massive BSA background after a simple UTLC step. This technique may have significant practical implications in actual detection of small molecules from complex food or clinical backgrounds.

  19. Using of scanner on the evaluation of pesticides mobility by thin-layer chromatography

    International Nuclear Information System (INIS)

    Tornisielo, V.L.; Costa, M.A.; Furlan, G.R.

    1995-01-01

    Knowledge of pesticide leaching potential is an essential information to preview environmental fate. The experiment confirms the possibility of using radiochromatogram scanning as a substitute for X-ray autoradiography, when Thin Layer Chromatografy (TLC) methodogy is used to determine mobility of a pesticide. Three types of soil from Sao Paulo state and five herbicides (metolachlor, asulan, simazing, 2,4-D and trifluralin), labeled with 14 C, were used. The radiochromatogram scanners permits a quick detection of the position of the radioactive spots to determine the Rf for each pesticide, while X-ray film has to be placed on the plate on the dark room for several days or weeks and then developed to detect spots, subsequently measure and calculate Rf. The results showed that the evaluation by scanner and X-ray were similar. Hence we conclude that the use of the scanner should be considered since this methodology is faster and as accurate as the X-ray methodology. (author). 4 refs, 1 fig, 2 tabs

  20. Peak broadening in paper chromatography and related techniques VI. The efficiency of various kinds of chromatography paper and thin-layer cellulose powder for the separation of amino acids

    NARCIS (Netherlands)

    Ligny, C.L. de; Kok, E.C.M.

    1968-01-01

    The efficiency of several chromatography papers and thin-layer cellulose powders for the separation of amino acids is investigated, using the minimum elution time for a given resolution as the criterion.

  1. Phosphorolytic activity of Escherichia coli glycyl-tRNA synthetase towards its cognate aminoacyl adenylate detected by 31P-NMR spectroscopy and thin-layer chromatography

    DEFF Research Database (Denmark)

    Led, Jens Jørgen; Switon, Werner K.; Jensen, Kaj Frank

    1983-01-01

    The catalytic activity of highly purified Escherichia coli glycyl-tRNA synthetase has been studied by 31P-NMR spectroscopy and thin-layer chromatography on poly(ethyleneimine)-cellulose. It was found that this synthetase, besides the activation of its cognate amino acid and the syntheses...

  2. Validation of an analytical method for the determination of spiramycin, virginiamycin and tylosin in feeding-stuffs bij thin-layer chromatography and bio-autography

    NARCIS (Netherlands)

    Vincent, U.; Gizzi, G.; Holst, von C.; Jong, de J.; Michard, J.

    2007-01-01

    An inter-laboratory validation was carried out to determine the performance characteristics of an analytical method based on thin-layer chromatography (TLC) coupled to microbiological detection (bio-autography) for screening feed samples for the presence of spiramycin, tylosin and virginiamycin.

  3. Stability indicating high performance thin-layer chromatographic method for simultaneous estimation of pantoprazole sodium and itopride hydrochloride in combined dosage form.

    Science.gov (United States)

    Bageshwar, Deepak; Khanvilkar, Vineeta; Kadam, Vilasrao

    2011-11-01

    A specific, precise and stability indicating high-performance thin-layer chromatographic method for simultaneous estimation of pantoprazole sodium and itopride hydrochloride in pharmaceutical formulations was developed and validated. The method employed TLC aluminium plates precoated with silica gel 60F 254 as the stationary phase. The solvent system consisted of methanol:water:ammonium acetate; 4.0:1.0:0.5 (v/v/v). This system was found to give compact and dense spots for both itopride hydrochloride ( R f value of 0.55±0.02) and pantoprazole sodium ( R f value of 0.85±0.04). Densitometric analysis of both drugs was carried out in the reflectance-absorbance mode at 289 nm. The linear regression analysis data for the calibration plots showed a good linear relationship with R 2 =0.9988±0.0012 in the concentration range of 100-400 ng for pantoprazole sodium. Also, the linear regression analysis data for the calibration plots showed a good linear relationship with R 2 =0.9990±0.0008 in the concentration range of 200-1200 ng for itopride hydrochloride. The method was validated for specificity, precision, robustness and recovery. Statistical analysis proves that the method is repeatable and selective for the estimation of both the said drugs. As the method could effectively separate the drug from its degradation products, it can be employed as a stability indicating method.

  4. Combined thin layer chromatography and gas chromatography with mass spectrometric analysis of lipid classes and fatty acids in malnourished polar bears (Ursus maritimus) which swam to Iceland.

    Science.gov (United States)

    Eibler, Dorothee; Krüger, Sabine; Skírnisson, Karl; Vetter, Walter

    2017-03-01

    Between 2008 and 2011, four polar bears (Ursus maritimus) from the Greenland population swam and/or drifted on ice to Iceland where they arrived in very poor body condition. Body fat resources in these animals were only between 0% and 10% of the body weight (usually 25%). Here we studied the lipid composition in different tissues (adipose tissue if available, liver, kidney and muscle). Lipid classes were determined by thin layer chromatography (TLC) and on-column gas chromatography with mass spectrometry (GC/MS). The fatty acid pattern of total lipids and free fatty acids was analyzed by GC/MS in selected ion monitoring (SIM) mode. Additionally, cholesteryl esters and native fatty acid methyl esters, initially detected as zones in thin layer chromatograms, were enriched by solid phase extraction and quantified by GC/MS. The ratio of free fatty acids to native fatty acid methyl esters could be correlated with the remained body lipids in the polar bears and thus may also serve as a marker for other starving animals or even for humans. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Radioactive preparations. Determination of radiochemical purity by thin-layer chromatography

    International Nuclear Information System (INIS)

    1986-01-01

    The standard sets the data which must be attached to every sample, and the equipment, chemicals and auxiliary substances used in the determination of radiochemical purity of substances by chromatography. Described are preparation of the sample, the procedure of sample deposition, the development, drying and detection of the radioactive preparation. The qualitative and quantitative assessment of the radiochromatogram is described as are the calculation of radiochemical purity and the determination of the reproducibility of measurement of radiochemical purity of radioactive preparations. (E.S.)

  6. Stable, microfabricated thin layer chromatography plates without volume distortion on patterned, carbon and Al₂O₃-primed carbon nanotube forests.

    Science.gov (United States)

    Jensen, David S; Kanyal, Supriya S; Gupta, Vipul; Vail, Michael A; Dadson, Andrew E; Engelhard, Mark; Vanfleet, Richard; Davis, Robert C; Linford, Matthew R

    2012-09-28

    Some of us recently described the fabrication of thin layer chromatography (TLC) plates from patterned carbon nanotube (CNT) forests via direct infiltration/coating of the CNTs by low pressure chemical vapor deposition (LPCVD) of silicon from SiH₄, followed by high temperature oxidation of the CNTs and Si. Herein we present an improved microfabrication process for the preparation of these TLC plates. First, a few nanometers of carbon and/or a thin film of Al₂O₃ is deposited on the CNTs. This method of priming the CNTs for subsequent depositions appears to be new. X-ray photoelectron spectroscopy confirms the presence of additional oxygen after carbon deposition. After priming, the plates are coated by rapid, conformal deposition of an inorganic material that does not require subsequent oxidation, i.e., by a fast pseudo atomic layer deposition (ψ-ALD) of SiO₂ from trimethylaluminum and tris(tert-butoxy)silanol. Unlike devices described previously, faithful reproduction of the features in the masks is still observed after oxidation. A bonded, amino phase on the resulting plates shows fast, highly efficient separations of fluorescent dyes (plate heights in the range of 1.6-7.7 μm). Extensive characterization of the new materials by TEM, SEM, EDAX, DRIFT, and XPS is reported. A substantially lower process temperature for the removal of the CNT scaffold is possible as a result of the already oxidized materials used. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. Complete Analysis of a Biologically Active Tetrapeptide: A Project Utilizing Thin-Layer Chromatography and Tandem Quadrupole Mass Spectrometry

    Science.gov (United States)

    Lefevre, Joseph W.; Dodsworth, David W.

    2000-04-01

    The biologically active tetrapeptide d-Ala-Gly-l-Phe-d-Leu ([des-Tyr1-d-Ala2-d-Leu5]enkephalin) was analyzed for its amino acid content and stereochemistry by normal and reversed-phase thin-layer chromatography (TLC), and its sequence was determined by tandem quadrupole mass spectrometry. The project involved sequential N-dansylation of a portion of the tetrapeptide, hydrolysis, isolation, and identification of the N-terminal amino acid as dansyl-alanine by comparison with standards using normal-phase TLC. A second portion of the tetrapeptide was hydrolyzed and the resulting four free amino acids were converted to their corresponding dansyl derivatives and purified by preparative normal-phase TLC. The three dansyl amino acids not identified previously were identified by TLC. The stereochemistry of each was determined by comparison with dansyl-dl-amino acid standards using reversed-phase TLC in the presence of ß-cyclodextrin, a chiral mobile phase additive. Finally, the correct amino acid sequence was determined by tandem quadrupole mass spectrometry. This project gives students valuable experience in microscale synthesis, both normal and reversed-phase TLC, stereochemical analysis, and mass spectrometry.

  8. Tuneable surface enhanced Raman spectroscopy hyphenated to chemically derivatized thin-layer chromatography plates for screening histamine in fish.

    Science.gov (United States)

    Xie, Zhengjun; Wang, Yang; Chen, Yisheng; Xu, Xueming; Jin, Zhengyu; Ding, Yunlian; Yang, Na; Wu, Fengfeng

    2017-09-01

    Reliable screening of histamine in fish was of urgent importance for food safety. This work presented a highly selective surface enhanced Raman spectroscopy (SERS) method mediated by thin-layer chromatography (TLC), which was tailored for identification and quantitation of histamine. Following separation and derivatization with fluram, plates were assayed with SERS, jointly using silver nanoparticle and NaCl. The latter dramatically suppressed the masking effect caused by excessive fluram throughout the plate, thus offering clear baseline and intensive Raman fingerprints specific to the analyte. Under optimized conditions, the usability of this method was validated by identifying the structural fingerprints of both targeted and unknown compounds in fish samples. Meanwhile, the quantitative results of this method agreed with those by an HPLC method officially suggested by EU for histamine determination. Showing remarkable cost-efficiency and user-friendliness, this facile TLC-SERS method was indeed screening-oriented and may be more attractive to controlling laboratories of limited resource. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Detection of adulterated copaiba (Copaifera multijuga Hayne oil-resins by refractive index and thin layer chromatography

    Directory of Open Access Journals (Sweden)

    Karol de S. Barbosa

    Full Text Available The refractive indices (RI of the eight samples of copaiba oils, collected for this study at RDS Tupé ranged from 1.50284 to 1.50786. The thin layer chromatography (TLC plates of these oils revealed with anisaldehyde-sulphuric acid reagent showed dark lilac stains with several small stains at low Rf and a large rounded stain at high Rf. On the other hand, the twelve copaiba oils purchased at local markets presented RI between 1.48176 and 1.50886, and the TLC plates, showed as general profile blue stains, with smaller superimposed stains at low Rf, bigger superimposed stains like elongated stain at high Rf and a colorless rounded stain at middle Rf. Among 12 purchased oils at local markets, a three oil-resins presented similar RI and TLC profile to those observed for collected copaiba oils; b six oils showed same RI and TLC profiles to those observed for soybean oil; c three samples presented RI near to those showed by copaiba oil-resin, however the TLC profile was near to profile observed for a prepared mixture soybean oil: copaiba oil, two samples with 3:1 proportion and one sample with 1:3 proportion. Therefore, the RI determination and the TLC profiles could be considered rapid and efficient procedures for detection of vegetal oil in the copaiba oil-resins.

  10. Re-evaluation of thin layer chromatography as an alternative method for the quantification of prostaglandins from rat Kupffer cells.

    Science.gov (United States)

    Pestel, Sabine; Jungermann, Kurt; Schieferdecker, Henrike L

    2005-01-01

    In contrast to conventionally used immunoassays, thin layer chromatography (TLC)--by prelabeling of cells with radioactive arachidonic acid (AA)--allows to differentiate between cellularly built and added prostanoids and thus to investigate feedback effects of prostanoids on their own release. PGD2, TXB2 and PGE2 released from zymosan-stimulated Kupffer cells were separated with distinct RF-values, corresponding to those of the pure substances. Quantification of PGD2 and PGE2 gave comparable results with TLC and immunoassays, but measurement in the presence of added prostanoids was only possible with TLC. Moreover TLC was superior to immunoassays in having a longer linear range while being comparably sensitive. Cellularly built TXB2 in its radioactively labeled form was not detectable by TLC. Inhibition of TXB2 release by externally added AA or technical artifacts were excluded, suggesting that the cellular AA-pools used for prostaglandin and thromboxane synthesis differ in their accessibility for added AA. Thus, TLC is a simple, sensitive and precise method for the quantification of cellularly built prostaglandins but not of thromboxane even in the presence of added prostanoids.

  11. Comparison of results for morphine urinalyses by radioimmunoassay and thin-layer chromatography in a narcotic clinic setting.

    Science.gov (United States)

    Kokoski, R J; Jain, M

    1975-03-01

    Radioimmunoassay (RIA) and thin-layer chromatography (TLC) were compared for morphine detection in an actual narcotic clinic setting. A choice of urines from all those screened by TLC allowed a critical comparison as to actual use or non-use of narcotic drugs, rather than a sampling at random in which the question of possible false positives or negatives cannot be conclusively answered. Although RIA is more sensitive than TLC, its advantage is apparent only in those cases where urine specimens are difficult to obtain frequently regularly or where the use of morphine is suspected by the positive identification of quinine in urine that was morphine-negative by TLC. In a selected group of negative and positive specimens chosen without conscious bias, the two methods gave consistently similar results, indicating that the modified TLC method provided a few or no false positives or negatives if the negatives were from those cases that were not positive anytime up to 3-4 days before urine collection. We conclude that RIA can be of significant value as a supplement to a TLC screening program, without sacrificing the many advantages that TLC has to offer.

  12. Validated High Performance Liquid Chromatography Method for ...

    African Journals Online (AJOL)

    Purpose: To develop a simple, rapid and sensitive high performance liquid ... response, tailing factor and resolution of six replicate injections was < 3 %. ... Cefadroxil monohydrate, Human plasma, Pharmacokinetics Bioequivalence ... Drug-free plasma was obtained from the local .... Influence of probenicid on the renal.

  13. Reversed-phase thin-layer chromatography of homologs of Antimycin-A and related derivatives

    Science.gov (United States)

    Abidi, Sharon L.

    1989-01-01

    Using a reversed-phase high-performance liquid chromatographic (HPLC) technique, a mixture of antimycins A was separated into eight hitherto unreported subcomponents, Ala, Alb, A2a, A2b, A3a, A3b, A4a, and A4b. Although a base-line resolution of the known four major antimycins Al, A2, A3, and A4 was readily achieved with mobile phases containing acetate buffers, the separation of the new antibiotic subcomponents was highly sensitive to variation in mobile phase conditions. The type and composition of organic modifiers, the nature of buffer salts, and the concentration of added electrolytes had profound effects on capacity factors, separation factors, and peak resolution values. Of the numerous chromatographic systems examined, a mobile phase consisting of methanol-water (70:30) and 0.005 M tetrabutylammonium phosphate at pH 3.0 yielded the most satisfactory results for the separation of the subcomponents. Reversed-phase gradient HPLC separation of the dansylated or methylated antibiotic compounds produced superior chromatographic characteristics and the presence of added electrolytes was not a critical factor for achieving separation. Differences in the chromatographic outcome between homologous and structural isomers were interpretated based on a differential solvophobic interaction rationale. Preparative reversed-phase HPLC under optimal conditions enabled isolation of pure samples of the methylated antimycin subcomponents for use in structural studies.

  14. Application of thin-layer chromatography of fat and oil, fatty acid and mineral oil for science education. Yushi, shibosan, koyu no hakuso chromatography to sono kyozaiteki katsuyo

    Energy Technology Data Exchange (ETDEWEB)

    Kawabuchi, K [Ehime Univ., Ehime (Japan); Takechi, Y; Morimoto, M [Ehime (Japan)

    1990-08-20

    In this article, with regard to application of thin-layer chromatography of fat and oil, fatty acid and mineral oil for science education, a study was made mainly on quality finding of n-hexane extracts provided in the law of prevention of water pollution, and separation of hydrolysis products of fat and oil. As a result, the existance of fat and oil, fatty acid and mineral oil in the enviromental water and the hydrolysis process of fat and oil could experimentally be confirmed. This is considered to be useful for studying teaching material on fat and oil. This report deals with the reagents and device, and specifies standard solution, thin-layer plate, spread solvent, spread tank and thermostat, and states, for its operation, the method using enzyme for hydrolysis of fat and oil and the method using potassium hydroxide. It also contemplates on R {sub f} values of fat and oil, fatty acid and mineral oil, etc., as well as spread of fat and oil, oils contained in n-hexane extracts and hydrolysis products of fat and oil. 6 refs., 4 figs., 1 tab.

  15. Feasibility of correlating separation of ternary mixtures of neutral analytes via thin layer chromatography with supercritical fluid chromatography in support of green flash separations.

    Science.gov (United States)

    Ashraf-Khorassani, M; Yan, Q; Akin, A; Riley, F; Aurigemma, C; Taylor, L T

    2015-10-30

    Method development for normal phase flash liquid chromatography traditionally employs preliminary screening using thin layer chromatography (TLC) with conventional solvents on bare silica. Extension to green flash chromatography via correlation of TLC migration results, with conventional polar/nonpolar liquid mixtures, and packed column supercritical fluid chromatography (SFC) retention times, via gradient elution on bare silica with a suite of carbon dioxide mobile phase modifiers, is reported. Feasibility of TLC/SFC correlation is individually described for eight ternary mixtures for a total of 24 neutral analytes. The experimental criteria for TLC/SFC correlation was assumed to be as follows: SFC/UV/MS retention (tR) increases among each of the three resolved mixture components; while, TLC migration (Rf) decreases among the same resolved mixture components. Successful correlation of TLC to SFC was observed for most of the polar organic solvents tested, with the best results observed via SFC on bare silica with methanol as the CO2 modifier and TLC on bare silica with a methanol/dichloromethane mixture. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Instant thin layer chromatography (ITLC) - the most true and convenient method for the determination of radiochemical purity of Tc-99m Labeled radiopharmaceuticals

    International Nuclear Information System (INIS)

    Anwar-Ul-Azim, M.; Ansari, M.I.H.

    2007-01-01

    Full text: Determination of radiochemical purity usually involves paper chromatography method or some column chromatography such as ion exchange or gel filtration technique. In this study, a modified chromatography method with thin-layer support called instant thin Layer chromatography (ITLC) was used for the measurement of radiochemical purity. The aim of the study was to present instant thin layer chromatography-Silica-Gel (ITLC-SG) technique as a fairly rapid , convenient and inexpensive system for the screening of radiochemical impurities of 99mTc-labeled radiopharmaceuticals resulting from physical , chemical and/or radiation decomposition. Materials and Methods: The experiment was carried out at the Institute of Nuclear Medicine and Ultrasound, Dhaka for a period of 10 months from August '05 to May '06. Radiochemical purity of seven types of 99mTc-labeled radiopharmaceuticals, namely 99mTc-DTPA,99mTc- MI I, 99m Tc - HDP, 99m Tc - DMSA, 99m Tc - Tetofosmin, 99mTc-Fyton and 99mTc-ECD were measured by the use of two solvent dual-instant thin layer chromatography systems. Commercially available instant thin layer chromatography plate ( 20cm x20cm) impregnated with silica gel (ITLC-SG: Merck, Germany) was used as a stationary phase for the measurement of radiochemical impurities.Acetone, Ethyl methyl ketone, chloroform and methanol were used to measure the amount of free pertechnetate in the labelled radiopharmaceuticals and 0.9% saline was used in all cases as solvent (Mobile phase) to measure the amount of the hydrolyzed-reduced technetium colloid. Results: The results showed that the measured radiochemical purity of 99mTc-DTPA, 99mTc-MIBI, 99mTc-HDP, 99mTc-DMSA, 99mTc-Tetofosmin, 99mTc-Fyton and 99mTc-ECD were acceptable for all the studied samples except two samples of 99mTc-MIBI, one sample of 99mTc-Fyton and two samples of 99mTc-ECD, where the measured radiochemical purity were not within the acceptable RCP limits of commonly used radiopharmaceuticals (RCP

  17. Quantitative determination of 3,4-methylenedioxymethamphetamine by thin-layer chromatography in ecstasy illicit pills in Tehran.

    Science.gov (United States)

    Shetab Boushehri, Seyed Vahid; Tamimi, Maryam; Kebriaeezadeh, Abbas

    2009-11-01

    3,4-Methylenedioxymethamphetamine (MDMA) is the major ingredient of ecstasy illicit pills. It is a hallucinogen, central nervous system stimulant, and serotonergic neurotoxin that strongly releases serotonin from serotonergic nerves terminals. Moreover, it releases norepinephrine and dopamine from nerves terminal, but to a lesser extent than serotonin. Poisoning and even death from abusing MDMA-containing ecstasy illicit pills among abusers is usual. Thus, quantitative determination of MDMA content of ecstasy illicit pills in illicit drug bazaar must be done regularly to find the most high dose ecstasy illicit pills and removing them from illicit drug bazaar. In the present study, MDMA contents of 13 most abundant ecstasy illicit pills were determined by quantitative thin-layer chromatography (TLC). Two procedures for quantitative determination of MDMA contents of ecstasy illicit pills by TLC were used: densitometric and so-called 'scraping off' methods. The former was done in a reflection mode at 285 nm and the latter was done by absorbance measurement of eluted scraped off spots. Limit of detection (LOD), considering signal-to-noise ratio (S/N) of 2, and limit of quantification (LOQ), regarding S/N of 10, of densitometric and scraping off methods were 0.40 microg, 1.20 microg, and 6.87 mug, 20.63 microg, respectively. Repeatabilities (within-laboratory error) of densitometric and scraping off methods were 0.5% and 3.6%, respectively. The results showed that the ecstasy illicit pills contained 24-124.5 mg and 23.9-122.2 mg MDMA by densitometric and scraping off methods, respectively.

  18. Thin layer chromatography fingerprint, antioxidant, and antibacterial activities of rhizomes, stems, and leaves of Curcuma aeruginosa Roxb.

    Science.gov (United States)

    Safitri, A.; Batubara, I.; Khumaida, N.

    2017-05-01

    Fingerprints of 5 temu hitam (Curcuma aeruginosa Roxb.) accessions (Malang, Cirebon, Kuningan 1, Bogor, and Liwa) were determined by thin-layer chromatography (TLC) and compared to fingerprints of turmeric (Curcuma longa L), temu putih (Curcuma zedoaria (Christm.) Roscoe), and temu lawak (Curcuma zanthorriza Roxb.). Maceration method with ethanol as the solvent was used for extraction. The eluent used for fingerprint by TLC was chloroform:dichloromethane (9:1v/v). Five accessions of temu hitam show similar fingerprint patterns, but different in band thickness. Temu hitam rhizomes have bands of curcuminoid (Rf 0.22, 0.10, 0.03), and characteristic bands of Rf 0.42, 0.27, and 0.77, which can be distinguished from turmeric and temu lawak and Rf 0.13, which is different from temu putih. Leaves and stems of temu hitam can be distinguished from temu putih, turmeric, and temu lawak at Rf 0.60. Rhizomes of all plants reveal strong antibacterial activity against Staphylococcus aureus and antioxidant activity on DPPH radicals than its corresponding stems and leaves. Antibacterial and antioxidant activities were determined by microdilution and TLC-bioautography. Antibacterial activity of rhizomes of Cirebon and Kuningan 1 accessions are higher than that of other accessions (MIC = 250 μg/mL MBC = 500 μg/mL, but lower as compared to that of temu lawak (MIC = 62.5 μg/mL, MBC = 250 μg/mL) and tetracycline (MIC = MBC = 15.63 μg/mL). Rhizome of Liwa accession exhibits the highest antioxidant activity (IC50 = 124.88 μg/mL) amongst all accessions, but lower than that of temu lawak (IC50 = 18.45 μg/mL), turmeric (IC50 = 18.82 μg/mL), and temu putih (IC50 = 94.35 μg/mL).

  19. Chromatographical analysis of phenolic acids in some species of Polygonum L. genus. Part 1 Qualitative analysis by two-dimensional thin layer chromatography (TLC

    Directory of Open Access Journals (Sweden)

    Helena D. Smolarz

    2014-01-01

    Full Text Available The Two-Dimensional Thin Layer Chromatography method has been used for the separation and identification of phenolic acids from six taxons of Pohygonum L. genus. The following acids were found: caffeic, p-coumaric, ferulic, p-hydroxybenzoic, m-hydroxybenzoic, vanillic, syringic, p-hydroxyphenylacetic, o-hydroxyphenylacetic, synapic, melillotic, salicylic, gentisic, elagic, gallic, chlorogenic, protocatechuic and homoprotocatechuic. Gallic, ferulic, vanillic, p-coumaric and p-hydroxybenzoic acids were isolated from herb Polygonum convolvulus L. using column chromatography.

  20. [In situ thin layer chromatography-fourier transform-surface-enhanced Raman spectrum study on ingredients of berberine].

    Science.gov (United States)

    Wang, Yuan; Guo, Zhan-sheng; Wang, Ying-feng; Wang, Song-ying; Ren, Gui-fen; Zhang, Xiang-lan; Han, Xiu-lan

    2002-10-01

    Surface Enhanced Raman Scattering (SERS) combined with Thin Layer Chromatography (TLC) has been used for studying characteristic spectrum of molecules in situ in micrograms samples. There are very few report for applying the SERS-TCL method in the study of the effective ingredients of Chinese traditional herbs. Coptis Chinensis France is an often-used clinic Chinese traditional medicine. Its main effective components include berberine and so on, which have antibiotic very wide and also have curative effect on improving the functions of heart vascular cycles. Therefore the concentrations of berberine are very important for the quality control of the medicine. In this work, the ethanol extract of Coptis Chinensis France was first separated by TLC, the SERS was then measure directly after dropping silver gel on the separated spots. The method can be used for the finger print analysis of the berberine. 3 microL of alcohol extract of Coptis Chinensis France with total alkaloids concentration of 1.0 mg.mL was placed on silicon GF254 TLC plate. The sample was separated by developing solvent of n bulanol-Acitic acid-H2O (7:2:1 V/V). The positions of berberine in the sample were confirmed by the standard alkaloid solutions. The Rf values for berberine are 0.29. The silver gel was used as surface enhanced substrate and placed on the separated berberine spots. FT-SERS was measured directly by a Nicolet FT-Raman 910 spectrometer. Berberine belong to isoquinoline alkaloids. His structure can be found in reference. The date of spectrum of berberine can be seen that the band at 1,396 cm-1 due to Ar-OCH3 deformation vibrations was greatly enhanced, indicating that the molecule was absorbed on silver gel strongly through lone-pair electron in Ar-OCH3. The ring stretching mode occurring around 1,548 cm-1 represents isoquinoline ring in the molecule. The band at 727 cm-1 due to CH (ring) deformation vibrations was also enhanced.

  1. The detection of some halogenated phenols and nitrophenols in thin-layer chromatography by means of bromine

    NARCIS (Netherlands)

    Tadema, G.; Batelaan, P.H.

    1968-01-01

    A method is described for the detection of halogeno- and nitro-phenols in sub-microgram quantities. Theses compounds are made visible by exposure of the developed thin layer plates to bromine vapour and subsequent spraying with an aqueous solution of potassium iodide or an ethanolic solution of

  2. Ultra-high Performance Liquid Chromatography in Steroid Analysis

    OpenAIRE

    Salonen, Fanny

    2017-01-01

    The latest version of liquid chromatography is ultra-high performance (or pressure) chromatography (UHPLC). In the technique, short and narrow-bore columns with particle sizes below 3 µm are used. The extremely high pressure used results in very short analysis times, excellent separation, and good resolution. This makes UHPLC a good choice for steroidal analysis. Steroids are a highly interesting area of study; they can be recognized as biomarkers for several diseases and are a relevant topic...

  3. Quantitative Thin-Layer Chromatography/Mass Spectrometry Analysis of Caffeine Using a Surface Sampling Probe Electrospray Ionization Tandem Mass Spectrometry System

    Energy Technology Data Exchange (ETDEWEB)

    Ford, Michael J [ORNL; Deibel, Michael A. [Earlham College; Tomkins, Bruce A [ORNL; Van Berkel, Gary J [ORNL

    2005-01-01

    Quantitative determination of caffeine on reversed-phase C8 thin-layer chromatography plates using a surface sampling electrospray ionization system with tandem mass spectrometry detection is reported. The thin-layer chromatography/electrospray tandem mass spectrometry method employed a deuterium-labeled caffeine internal standard and selected reaction monitoring detection. Up to nine parallel caffeine bands on a single plate were sampled in a single surface scanning experiment requiring 35 min at a surface scan rate of 44 {mu}m/s. A reversed-phase HPLC/UV caffeine assay was developed in parallel to assess the mass spectrometry method performance. Limits of detection for the HPLC/UV and thin-layer chromatography/electrospray tandem mass spectrometry methods determined from the calibration curve statistics were 0.20 ng injected (0.50 {mu}L) and 1.0 ng spotted on the plate, respectively. Spike recoveries with standards and real samples ranged between 97 and 106% for both methods. The caffeine content of three diet soft drinks (Diet Coke, Diet Cherry Coke, Diet Pepsi) and three diet sport drinks (Diet Turbo Tea, Speed Stack Grape, Speed Stack Fruit Punch) was measured. The HPLC/UV and mass spectrometry determinations were in general agreement, and these values were consistent with the quoted values for two of the three diet colas. In the case of Diet Cherry Coke and the diet sports drinks, the determined caffeine amounts using both methods were consistently higher (by 8% or more) than the literature values.

  4. Simplified and rapid method for extraction of ergosterol from natural samples and detection with quantitative and semi-quantitative methods using thin-layer chromatography

    OpenAIRE

    Larsen, Cand.scient Thomas; Ravn, Senior scientist Helle; Axelsen, Senior Scientist Jørgen

    2004-01-01

    A new and simplified method for extraction of ergosterol (ergoste-5,7,22-trien-3-beta-ol) from fungi in soil and litter was developed using pre-soaking extraction and paraffin oil for recovery. Recoveries of ergosterol were in the range of 94 - 100% depending on the solvent to oil ratio. Extraction efficiencies equal to heat-assisted extraction treatments were obtained with pre-soaked extraction. Ergosterol was detected with thin-layer chromatography (TLC) using fluorodensitometry with a quan...

  5. Óleo essencial de limão no ensino da cromatografia em camada delgada Lime essential oil in the teaching of thin layer chromatography

    Directory of Open Access Journals (Sweden)

    Rosaly S. Silva

    2009-01-01

    Full Text Available This paper describes a simple experiment employing the essential oil of limes which can be applied in undergraduate organic chemistry laboratory classes for the teaching of thin layer chromatography (TLC. The experiment consists in submit lime peel oil to TLC separation employing hexane and dichloromethane as the eluents and five different systems for visualization of the chromatogram. In one experiment it is possible to teach the different variables of the TLC technique. This experiment may also be performed following vapor distillation and liquid-liquid extraction technique in experimental classes.

  6. Determination of Caffeine in Beverages by High Performance Liquid Chromatography.

    Science.gov (United States)

    DiNunzio, James E.

    1985-01-01

    Describes the equipment, procedures, and results for the determination of caffeine in beverages by high performance liquid chromatography. The method is simple, fast, accurate, and, because sample preparation is minimal, it is well suited for use in a teaching laboratory. (JN)

  7. Mallow carotenoids determined by high-performance liquid chromatography

    Science.gov (United States)

    Mallow (corchorus olitorius) is a green vegetable, which is widely consumed either fresh or dry by Middle East population. This study was carried out to determine the contents of major carotenoids quantitatively in mallow, by using a High Performance Liquid Chromatography (HPLC) equipped with a Bis...

  8. High-Performance Liquid Chromatography-Mass Spectrometry.

    Science.gov (United States)

    Vestal, Marvin L.

    1984-01-01

    Reviews techniques for online coupling of high-performance liquid chromatography with mass spectrometry, emphasizing those suitable for application to nonvolatile samples. Also summarizes the present status, strengths, and weaknesses of various techniques and discusses potential applications of recently developed techniques for combined liquid…

  9. Monitoring aged reversed-phase high performance liquid chromatography columns

    NARCIS (Netherlands)

    Bolck, A; Smilde, AK; Bruins, CHP

    1999-01-01

    In this paper, a new approach for the quality assessment of routinely used reversed-phase high performance liquid chromatography columns is presented. A used column is not directly considered deteriorated when changes in retention occur. If attention is paid to the type and magnitude of the changes,

  10. Ultra high performance liquid chromatography of seized drugs

    NARCIS (Netherlands)

    Lurie, I.S.

    2010-01-01

    The primary goal of this thesis is to investigate the use of ultra high performance liquid chromatography (UHPLC) for the analysis of seized drugs. This goal was largely achieved and significant progress was made in achieving improved separation and detection of drugs of forensic interest.

  11. Quantification of Tea Flavonoids by High Performance Liquid Chromatography

    Science.gov (United States)

    Freeman, Jessica D.; Niemeyer, Emily D.

    2008-01-01

    We have developed a laboratory experiment that uses high performance liquid chromatography (HPLC) to quantify flavonoid levels in a variety of commercial teas. Specifically, this experiment analyzes a group of flavonoids known as catechins, plant-derived polyphenolic compounds commonly found in many foods and beverages, including green and black…

  12. Radioactivity monitor for high-performance liquid chromatography

    International Nuclear Information System (INIS)

    Reeve, D.R.; Crozier, A.

    1977-01-01

    The coupling of a homogeneous radioactivity monitor to a liquid chromatograph involves compromises between the sensitivity of the monitor and the resolution and speed of analysis of the chromatograph. The theoretical relationships between these parameters are considered and expressions derived which make it possible to calculate suitable monitor operating conditions for most types of high-performance liquid chromatography

  13. Thin Layer Chromatography-Bioautography and Gas Chromatography-Mass Spectrometry of Antimicrobial Leaf Extracts from Philippine Piper betle L. against Multidrug-Resistant Bacteria

    Directory of Open Access Journals (Sweden)

    Demetrio L. Valle

    2016-01-01

    Full Text Available This study isolated and identified the antimicrobial compounds of Philippine Piper betle L. leaf ethanol extracts by thin layer chromatography- (TLC- bioautography and gas chromatography-mass spectrometry (GC-MS. Initially, TLC separation of the leaf ethanol extracts provided a maximum of eight compounds with Rf values of 0.92, 0.86, 0.76, 0.53, 0.40, 0.25, 0.13, and 0.013, best visualized when inspected under UV 366 nm. Agar-overlay bioautography of the isolated compounds demonstrated two spots with Rf values of 0.86 and 0.13 showing inhibitory activities against two Gram-positive multidrug-resistant (MDR bacteria, namely, methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus. The compound with an Rf value of 0.86 also possessed inhibitory activity against Gram-negative MDR bacteria, namely, carbapenem-resistant Enterobacteriaceae-Klebsiella pneumoniae and metallo-β-lactamase-producing Acinetobacter baumannii. GC-MS was performed to identify the semivolatile and volatile compounds present in the leaf ethanol extracts. Six compounds were identified, four of which are new compounds that have not been mentioned in the medical literature. The chemical compounds isolated include ethyl diazoacetate, tris(trifluoromethylphosphine, heptafluorobutyrate, 3-fluoro-2-propynenitrite, 4-(2-propenylphenol, and eugenol. The results of this study could lead to the development of novel therapeutic agents capable of dealing with specific diseases that either have weakened reaction or are currently not responsive to existing drugs.

  14. Thin Layer Chromatography-Bioautography and Gas Chromatography-Mass Spectrometry of Antimicrobial Leaf Extracts from Philippine Piper betle L. against Multidrug-Resistant Bacteria.

    Science.gov (United States)

    Valle, Demetrio L; Puzon, Juliana Janet M; Cabrera, Esperanza C; Rivera, Windell L

    2016-01-01

    This study isolated and identified the antimicrobial compounds of Philippine Piper betle L. leaf ethanol extracts by thin layer chromatography- (TLC-) bioautography and gas chromatography-mass spectrometry (GC-MS). Initially, TLC separation of the leaf ethanol extracts provided a maximum of eight compounds with R f values of 0.92, 0.86, 0.76, 0.53, 0.40, 0.25, 0.13, and 0.013, best visualized when inspected under UV 366 nm. Agar-overlay bioautography of the isolated compounds demonstrated two spots with R f values of 0.86 and 0.13 showing inhibitory activities against two Gram-positive multidrug-resistant (MDR) bacteria, namely, methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus. The compound with an R f value of 0.86 also possessed inhibitory activity against Gram-negative MDR bacteria, namely, carbapenem-resistant Enterobacteriaceae-Klebsiella pneumoniae and metallo-β-lactamase-producing Acinetobacter baumannii. GC-MS was performed to identify the semivolatile and volatile compounds present in the leaf ethanol extracts. Six compounds were identified, four of which are new compounds that have not been mentioned in the medical literature. The chemical compounds isolated include ethyl diazoacetate, tris(trifluoromethyl)phosphine, heptafluorobutyrate, 3-fluoro-2-propynenitrite, 4-(2-propenyl)phenol, and eugenol. The results of this study could lead to the development of novel therapeutic agents capable of dealing with specific diseases that either have weakened reaction or are currently not responsive to existing drugs.

  15. Comparison of ultra high performance supercritical fluid chromatography, ultra high performance liquid chromatography, and gas chromatography for the separation of synthetic cathinones.

    Science.gov (United States)

    Carnes, Stephanie; O'Brien, Stacey; Szewczak, Angelica; Tremeau-Cayel, Lauriane; Rowe, Walter F; McCord, Bruce; Lurie, Ira S

    2017-09-01

    A comparison of ultra high performance supercritical fluid chromatography, ultra high performance liquid chromatography, and gas chromatography for the separation of synthetic cathinones has been conducted. Nine different mixtures of bath salts were analyzed in this study. The three different chromatographic techniques were examined using a general set of controlled synthetic cathinones as well as a variety of other synthetic cathinones that exist as positional isomers. Overall 35 different synthetic cathinones were analyzed. A variety of column types and chromatographic modes were examined for developing each separation. For the ultra high performance supercritical fluid chromatography separations, analyses were performed using a series of Torus and Trefoil columns with either ammonium formate or ammonium hydroxide as additives, and methanol, ethanol or isopropanol organic solvents as modifiers. Ultra high performance liquid chromatographic separations were performed in both reversed phase and hydrophilic interaction chromatographic modes using SPP C18 and SPP HILIC columns. Gas chromatography separations were performed using an Elite-5MS capillary column. The orthogonality of ultra high performance supercritical fluid chromatography, ultra high performance liquid chromatography, and gas chromatography was examined using principal component analysis. For the best overall separation of synthetic cathinones, the use of ultra high performance supercritical fluid chromatography in combination with gas chromatography is recommended. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. [A thin-layer chromatography method for determining the styrene metabolites mandelic and phenylglyoxylic acid in urine].

    Science.gov (United States)

    Gartzke, J; Burck, D

    1989-06-01

    A thin-layer chromatographic method is described for the determination of mandelic and phenyglyoxillic acid on silicagel (Silufol UV 254) after extraction from urine of styrene exposed workers. The quantitative determination was performed after eluting the spots. Phenylglyoxilic acid was measured at 255 nm and mandelic acid by derivative spectroscopically estimation of the .CH(OH).COOH -chromophore at 217 nm or by a three-wavelength mode, respectively. The recovery in urine was 80-104% for phenylglyoxilic acid and 99-105% for mandelic acid.

  17. Simultaneous determination of atorvastatin calcium and ramipril in capsule dosage forms by high-performance liquid chromatography and high-performance thin layer chromatography.

    Science.gov (United States)

    Panchal, Hiral J; Suhagia, Bhanubhai N

    2010-01-01

    Two simple and accurate methods to determine atorvastatin calcium and ramipril in capsule dosage forms were developed and validated using HPLC and HPTLC. The HPLC separation was achieved on a Phenomenex Luna C18 column (250 x 4.6 mm id, 5 microm) in the isocratic mode using 0.1% phosphoric acid-acetonitrile (38 + 62, v/v), pH 3.5 +/- 0.05, mobile phase at a flow rate of 1 ml/min. The retention times were 6.42 and 2.86 min for atorvastatin calcium and ramipril, respectively. Quantification was achieved with a photodiode array detector set at 210 nm over the concentration range of 0.5-5 microg/mL for each, with mean recoveries (at three concentration levels) of 100.06 +/- 0.49% and 99.95 +/- 0.63% RSD for atorvastatin calcium and ramipril, respectively. The HPTLC separation was achieved on silica gel 60 F254 HPTLC plates using methanol-benzene-glacial acetic acid (19.6 + 80.0 + 0.4, v/v/v) as the mobile phase. The Rf values were 0.40 and 0.20 for atorvastatin calcium and ramipril, respectively. Quantification was achieved with UV densitometry at 210 nm over the concentration range of 50-500 ng/spot for each, with mean recoveries (at three concentration levels) of 99.98 +/- 0.75% and 99.87 +/- 0.83% RSD for atorvastatin calcium and ramipril, respectively. Both methods were validated according to International Conference on Harmonization guidelines and found to be simple, specific, accurate, precise, and robust. The mean assay percentages for atorvastatin calcium and ramipril were 99.90 and 99.55% for HPLC and 99.91 and 99.47% for HPTLC, respectively. The methods were successfully applied for the determination of atorvastatin calcium and ramipril in capsule dosage forms without any interference from common excipients.

  18. Rapid, simple, and highly sensitive analysis of drugs in biological samples using thin-layer chromatography coupled with matrix-assisted laser desorption/ionization mass spectrometry.

    Science.gov (United States)

    Kuwayama, Kenji; Tsujikawa, Kenji; Miyaguchi, Hajime; Kanamori, Tatsuyuki; Iwata, Yuko T; Inoue, Hiroyuki

    2012-01-01

    Rapid and precise identification of toxic substances is necessary for urgent diagnosis and treatment of poisoning cases and for establishing the cause of death in postmortem examinations. However, identification of compounds in biological samples using gas chromatography and liquid chromatography coupled with mass spectrometry entails time-consuming and labor-intensive sample preparations. In this study, we examined a simple preparation and highly sensitive analysis of drugs in biological samples such as urine, plasma, and organs using thin-layer chromatography coupled with matrix-assisted laser desorption/ionization mass spectrometry (TLC/MALDI/MS). When the urine containing 3,4-methylenedioxymethamphetamine (MDMA) without sample dilution was spotted on a thin-layer chromatography (TLC) plate and was analyzed by TLC/MALDI/MS, the detection limit of the MDMA spot was 0.05 ng/spot. The value was the same as that in aqueous solution spotted on a stainless steel plate. All the 11 psychotropic compounds tested (MDMA, 4-hydroxy-3-methoxymethamphetamine, 3,4-methylenedioxyamphetamine, methamphetamine, p-hydroxymethamphetamine, amphetamine, ketamine, caffeine, chlorpromazine, triazolam, and morphine) on a TLC plate were detected at levels of 0.05-5 ng, and the type (layer thickness and fluorescence) of TLC plate did not affect detection sensitivity. In addition, when rat liver homogenate obtained after MDMA administration (10 mg/kg) was spotted on a TLC plate, MDMA and its main metabolites were identified using TLC/MALDI/MS, and the spots on a TLC plate were visualized by MALDI/imaging MS. The total analytical time from spotting of intact biological samples to the output of analytical results was within 30 min. TLC/MALDI/MS enabled rapid, simple, and highly sensitive analysis of drugs from intact biological samples and crude extracts. Accordingly, this method could be applied to rapid drug screening and precise identification of toxic substances in poisoning cases and

  19. Combined high-performance liquid chromatography-radioimmunoassay for cytokinins

    International Nuclear Information System (INIS)

    MacDonald, E.M.S.; Akiyoshi, D.E.; Morris, R.O.

    1981-01-01

    The cytokinins isopentenyladenosine and ribosylzeatin were conjugated to bovine serum albumin and the conjugates used to raise antisera in rabbits. The resulting antisera had high specificity towards the cytokinin haptens and low cross-reactivity towards other purines. They were used as the basis for a radioimmunoassay for cytokinins, which, when applied in conjunction with high-performance liquid chromatography, allowed rapid and sensitive (to the picogram range) estimation and identification of multiple cytokinins from natural plant and bacterial sources. (orig.)

  20. High performance liquid chromatography in studies of radiolabeled antibodies

    International Nuclear Information System (INIS)

    Hnatowich, D.J.

    1986-01-01

    High performance liquid chromatography (HPLC) as applied to the separation of antibodies displays the same advantages as in its other applications, namely good resolution accompanied by fast analysis. It is therefore not surprising that many HPLC columns designed for use with antibodies and other proteins are now available commercially. The properties of proteins which provide the separation are size, hydrophobicity, charge and affinity. The features of each are discussed. (author)

  1. Preconcentration of rare earth elements from rocks by thin-layer chromatography and their neutron-activation determination

    International Nuclear Information System (INIS)

    Ryabukhin, V.A.; Volynets, M.P.; Myasoedov, B.F.

    1990-01-01

    Conditions were studied for separation of rare earths and accompanying elements in rocks on Fixion 50x8 thin-layer plates using solutions of oxalic acid and ammonium chloride in ammonia medium. A simple technique was developed for TLC proconcentration of rare earths followed by gamma spectrometric analysis of the irradiated fractions that enabled to determine 8-10 elements in samples with a mass of up to 30 mg. The limits of detection (μg/g) were 0.05 (Eu), 0.1 (Sm), 0.2 (Tb), 0.3 (Yb), 0.4 (La), 1.0 (Tu), 2.0 (Ce), 10 (Nd). The relative standard deviation was 0.05-0.20 at element levels 5-10 times as high as the detection limits

  2. Chromatography.

    Science.gov (United States)

    Brantley, L. Reed, Sr.; Demanche, Edna L.; Klemm, E. Barbara; Kyselka, Will; Phillips, Edwin A.; Pottenger, Francis M.; Yamamoto, Karen N.; Young, Donald B.

    This booklet presents some activities on chromatography. Directions for preparing leaf pigment extracts using alcohol are given, and paper chromatography and thin-layer chromatography are described as modifications of the basic principles of chromatography. (KHR)

  3. Thin-layer chromatography of 49 metal ions on stannic antimonate in aqueous and mixed solvent systems containing dimethylsulfoxide: quantitative separation of uranium from numerous metal ions

    International Nuclear Information System (INIS)

    Qureshi, M.; Varshney, K.G.; Rajput, R.P.S.

    1976-01-01

    Thin-layer chromatography of 40 metal ions in 31 aqueous and mixed solvent systems has been performed on stannic antimonate ion-exchange material. Dimethylsulfoxide has been utilized to resolve such binary mixtures as La 3+ and Ce 3+ from Pr 3+ , Nd 3+ , and Sm 3+ ; VO 2+ from Ti 4+ , Nb 5+ , and Ta 5+ ; Ga 3+ from In 3+ , Tl + , and Y 3+ ; Fe 3+ from VO 2+ ; and Mg 2+ from Al 3+ . Quantitative separation of 200 to 800 μg U from its binary mixtures and from the synthetic mixtures containing Mg 2+ , Bi 3+ , Fe 3+ , Th 4+ , Ce 4+ , Cr 3+ , Zr 4+ , Hf 4+ , Ti 4+ Mn 2+ , Cu 2+ , Ce 3+ , In 3+ , Y 3+ , Ca 2+ , Co 2+ , Tl + , Nb 5+ , and Ag + has been obtained

  4. Composite glycerol/graphite/aromatic acid matrices for thin-layer chromatography/matrix-assisted laser desorption/ionization mass spectrometry of heterocyclic compounds.

    Science.gov (United States)

    Esparza, Cesar; Borisov, R S; Varlamov, A V; Zaikin, V G

    2016-10-28

    New composite matrices have been suggested for the analysis of mixtures of different synthetic organic compounds (N-containing heterocycles and erectile dysfunction drugs) by thin layer chromatography/matrix-assisted laser desorption ionization time-of-flight mass spectrometry (TLC/MALDI-TOF). Different mixtures of classical MALDI matrices and graphite particles dispersed in glycerol were used for the registration of MALDI mass spectra directly from TLC plates after analytes separation. In most of cases, the mass spectra possessed [M+H] + ions; however, for some analytes only [M+Na] + and [M+K] + ions were observed. These ions have been used to generate visualized TLC chromatograms. The described approach increases the desorption/ionization efficiencies of analytes separated by TLC, prevent spot blurring, simplifies and decrease time for sample preparation. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Forensic examination of ink by high-performance thin layer chromatography--the United States Secret Service Digital Ink Library.

    Science.gov (United States)

    Neumann, Cedric; Ramotowski, Robert; Genessay, Thibault

    2011-05-13

    Forensic examinations of ink have been performed since the beginning of the 20th century. Since the 1960s, the International Ink Library, maintained by the United States Secret Service, has supported those analyses. Until 2009, the search and identification of inks were essentially performed manually. This paper describes the results of a project designed to improve ink samples' analytical and search processes. The project focused on the development of improved standardization procedures to ensure the best possible reproducibility between analyses run on different HPTLC plates. The successful implementation of this new calibration method enabled the development of mathematical algorithms and of a software package to complement the existing ink library. Copyright © 2010 Elsevier B.V. All rights reserved.

  6. Densitometric Validation and Optimisation of Polyphenols in Ocimum sanctum Linn by High Performance Thin-layer Chromatography.

    Science.gov (United States)

    U K, Ilyas; Katare, Deepshikha P; Aeri, Vidhu

    2015-01-01

    Ocimum sanctum Linn (Sanskrit: Tulasi; family: Libiaceae), popularly known as holy basil or Ocimum teinufolium, is found throughout the semitropical and tropical parts of India. In Ayurveda, Tulasi has been well known for its therapeutic potentials. To optimise and develop a standard method to quantify seven polyphenols simultaneously by HPTLC. A three-level factor Box-Behnken statistical design was used for optimisation, where extraction time (min), temperature (°C) and methanol:water ratio (% v/v) are the independent variables with polyphenols as the dependent variable. The separation was archived on a silica-gel 60 F254 HPTLC plate using toluene:ethyl acetate:formic acid:methanol (3:3:0.8:0.2 v/v) as the mobile phase. Densitometric analysis of polyphenols was carried out in the absorbance mode at 366 nm. The quantification of polyphenols was carried out based on peak area with a linear calibration curve at concentration ranges of 60-240, 20-200, 100-1600, 40-200, 200-1400, 10-160, 200-1400, 100-5000 ng/band for caffeic acid, ellagic acid, rutin, kaempferol, catechin, quercetin, eupalitin and epicatechin respectively. The method was validated for peak purity, precision, accuracy, limit of detection (LOD) and quantification (LOQ). Method specificity was confirmed using the retention factor value and visible spectra correlation of marker compounds. A validated HPTLC method was newly developed for simultaneous quantification of seven polyphenols in an Ayurvedic preparation of O. sanctum. The proposed method is simple, precise, specific, accurate, cost-effective, less time consuming and has the ability to separate the polyphenols from other constituents. Copyright © 2015 John Wiley & Sons, Ltd.

  7. Phospholipid composition of cell-derived microparticles determined by one-dimensional high-performance thin-layer chromatography

    NARCIS (Netherlands)

    Weerheim, A. M.; Kolb, A. M.; Sturk, A.; Nieuwland, R.

    2002-01-01

    Microparticles in the circulation activate the coagulation system and may activate the complement system via C-reactive protein upon conversion of membrane phospholipids by phospholipases. We developed a sensitive and reproducible method to determine the phospholipid composition of microparticles.

  8. Hydrazine Determination in Sludge Samples by High Performance Liquid Chromatography

    Energy Technology Data Exchange (ETDEWEB)

    G. Elias; G. A. Park

    2006-02-01

    A high-performance liquid chromatographic method using ultraviolet (UV) detection was developed to detect and quantify hydrazine in a variety of environmental matrices. The method was developed primarily for sludge samples, but it is also applicable to soil and water samples. The hydrazine in the matrices was derivatized to their hydrazones with benzaldehyde. The derivatized hydrazones were separated using high performance liquid chromatography (HPLC) with a reversed-phase C-18 column in an isocratic mode with methanol-water (95:5, v/v), and detected with UV detection at 313 nm. The detection limit (25 ml) for the new analytical method is 0.0067 mg ml-1of hydrazine. Hydrazine showed low recovery in soil samples because components in soil oxidized hydrazine. Sludge samples that contained relatively high soil content also showed lower recovery. The technique is relatively simple and cost-effective, and is applicable for hydrazine analysis in different environmental matrices.

  9. Development and Validation of High Performance Thin-Layer Chromatographic Method for Determination of α-Mangostin in Fruit Pericarp of Mangosteen Plant (Garcinia mangostana L. using Ultraviolet – Visible Detection

    Directory of Open Access Journals (Sweden)

    Himanshu Misra

    2009-10-01

    Full Text Available A simple, fast and precise quantitative high performance thin-layer chromatographic method has been developed for quantitative estimation of α-mangostin in fruit pericarp of Garcinia mangostana L. (Hypericaceae. Best solvent for extraction of a-mangostin optimized after screening with five solvents under same conditions using hot solid-liquid extraction through soxhlet apparatus. Methanol and chloroform gave highest and second highest recovery of a-mangostin, respectively. Plates were developed in chloroform-methanol in the ratio of 27-3 (v/v. Post-chromatographic derivatization performed using anisaldehyde-sulphuric acid reagent and scanned at 382 nm in ultraviolet-visible mode. The developed method was found to be linear in the range 1.0 to 5.0 mg spot-1, limits of detection and quantitation were 150 and 450 ng spot-1. The developed method was validated in terms of system suitability, specificity and robustness.

  10. Combined computational-experimental approach to predict blood-brain barrier (BBB) permeation based on "green" salting-out thin layer chromatography supported by simple molecular descriptors.

    Science.gov (United States)

    Ciura, Krzesimir; Belka, Mariusz; Kawczak, Piotr; Bączek, Tomasz; Markuszewski, Michał J; Nowakowska, Joanna

    2017-09-05

    The objective of this paper is to build QSRR/QSAR model for predicting the blood-brain barrier (BBB) permeability. The obtained models are based on salting-out thin layer chromatography (SOTLC) constants and calculated molecular descriptors. Among chromatographic methods SOTLC was chosen, since the mobile phases are free of organic solvent. As consequences, there are less toxic, and have lower environmental impact compared to classical reserved phases liquid chromatography (RPLC). During the study three stationary phase silica gel, cellulose plates and neutral aluminum oxide were examined. The model set of solutes presents a wide range of log BB values, containing compounds which cross the BBB readily and molecules poorly distributed to the brain including drugs acting on the nervous system as well as peripheral acting drugs. Additionally, the comparison of three regression models: multiple linear regression (MLR), partial least-squares (PLS) and orthogonal partial least squares (OPLS) were performed. The designed QSRR/QSAR models could be useful to predict BBB of systematically synthesized newly compounds in the drug development pipeline and are attractive alternatives of time-consuming and demanding directed methods for log BB measurement. The study also shown that among several regression techniques, significant differences can be obtained in models performance, measured by R 2 and Q 2 , hence it is strongly suggested to evaluate all available options as MLR, PLS and OPLS. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Highly sensitive on-site detection of drugs adulterated in botanical dietary supplements using thin layer chromatography combined with dynamic surface enhanced Raman spectroscopy.

    Science.gov (United States)

    Fang, Fang; Qi, Yunpeng; Lu, Feng; Yang, Liangbao

    2016-01-01

    The phenomenon of botanical dietary supplements (BDS) doped with illegal adulterants has become a serious problem all over the world, which could cause great threat to human's health. Therefore, it is of great value to identify BDS. Herein, we put forward a highly sensitive method for on-site detection of antitussive and antiasthmatic drugs adulterated in BDS using thin layer chromatography (TLC) combined with dynamic surface enhanced Raman spectroscopy (DSERS). Adulterants in BDS were separated on a TLC plate and located under UV illumination. Then DSERS detection was performed using a portable Raman spectrometer with 50% glycerol silver colloid serving as DSERS active substrate. Here, the effects of different solvents on detection efficacy were evaluated using phenformin hydrochloride (PHE) as a probe. It was shown that 50% glycerol resulted in higher SERS enhancement and relatively higher stability. Moreover, practical application of this novel TLC-DSERS method was demonstrated with rapid analysis of real BDS samples and one sample adulterated with benproperine phosphate (BEN) was found. Furthermore, the obtained result was verified by ultra performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-QTOF/MS). The sensitivity of the TLC-DSERS technique is 1-2 orders of magnitude higher than that of TLC-SERS technique. The results turned out that this combined method would have good prospects for on-site and sensitive detection of adulterated BDS. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Characterization of Mycobacterium paratuberculosis by gas-liquid and thin-layer chromatography and rapid demonstration of mycobactin dependence using radiometric methods

    International Nuclear Information System (INIS)

    Damato, J.J.; Knisley, C.; Collins, M.T.

    1987-01-01

    Thirty-six Mycobacterium paratuberculosis isolates of bovine, caprine, and ovine origins were evaluated by using gas-liquid chromatography (GLC), thin-layer chromatography (TLC), and BACTEC 7H12 Middlebrook TB medium in an effort to more rapidly differentiate this group of organisms from other mycobacteria. Bacterial suspensions (0.1 ml) were inoculated by syringe into 7H12 broth containing 2 micrograms of mycobactin P per ml and control broth without mycobactin P. Cultures were incubated at 37 0 C and read daily with a BACTEC Model 301. After 8 days of incubation, the growth index readings for the test broths containing mycobactin P were twice those of the control broths without mycobactin P. Sixty-five isolates of mycobacteria other than M. paratuberculosis were also examined. No difference was noted between the growth index readings of control and mycobactin-containing broths. Except for Mycobacterium avium-Mycobacterium intracellulare, TLC studies differentiated M. paratuberculosis from the other mycobacterial species tested. The GLC data reveal that all M. paratuberculosis isolates had a distinctive peak (14A) which was not found among M. avium-M. intracellulare complex organisms. These data indicate that 7H12 radiometric broth was able to rapidly demonstrate the mycobactin dependence of M. paratuberculosis and GLC and TLC procedures were capable of rapidly differentiating this organism from the other mycobacteria studied

  13. Extraction of oxytetracycline starting from the meats of chickens and identifications by thin layer chromatography; Extraction de l'oxytetracycline dans les viandes des poulet et identification par chromatographie sur couche mince

    Energy Technology Data Exchange (ETDEWEB)

    Elghozzi, Amira [Institut Superieur de Biotechnologie de Sidi Thabet (Tunisia)

    2007-07-01

    Use of antibiotics in the poultry also suspected as one of the cause of the emergence of resistance to antibiotics of some bacteria what causes risks on human health continuation of persistence of the residues in the foodstuffs. We were interested in detected the presence qualitatively of oxytetracyclin in samples of muscle and liver of chicken by the use of the techniques of thin layer chromatography. Although, These results are satisfactory, it is always necessary to prevent the dangers which cause the risks of the residues of oxytetracycline on health. (Author). 20 refs.

  14. Fractionation of Cardiolite for clinical use: comparative evaluation of radiochemical purity by thin-layer and paper chromatography of 99mTc-hexakis-2-methoxy-isobutyl-isonitrile complex

    International Nuclear Information System (INIS)

    Albernaz, M.S.; Pantoja, M.R.; Martinho, M.J.R.; Fonseca, M.L.B.; Bernardo-Filho, Mario

    1996-01-01

    Twenty-five vials of the previous divided Cardiolite (technetium-99m-hexakis-2-metoxi-isobutyl-isonitrile) were evaluated immediately after labeling with technetium-99m and in 15 them the specific activity assurance was checked at hourly intervals up to 3 hours. Quality control assessment comprised instant thin-layer chromatography, as suggested by the manufacturer, compared to paper chromatography which is widely available and less expensive than the former. Among the 25 bottles, 21 had a radiochemical purity higher than 89%. The redistribution of the Cardiolite in small doses vacuum containers was a practical and reliable technique. The technetium-99m-isonitrile was a stable radiochemical compound thought the three hours period of observation and proved adequate for clinical use. When thin-layer and paper chromatography were compared there was no significant difference regarding radiochemical purity. (author)

  15. Building blocks for the development of an interface for high-throughput thin layer chromatography/ambient mass spectrometric analysis: a green methodology.

    Science.gov (United States)

    Cheng, Sy-Chyi; Huang, Min-Zong; Wu, Li-Chieh; Chou, Chih-Chiang; Cheng, Chu-Nian; Jhang, Siou-Sian; Shiea, Jentaie

    2012-07-17

    Interfacing thin layer chromatography (TLC) with ambient mass spectrometry (AMS) has been an important area of analytical chemistry because of its capability to rapidly separate and characterize the chemical compounds. In this study, we have developed a high-throughput TLC-AMS system using building blocks to deal, deliver, and collect the TLC plate through an electrospray-assisted laser desorption ionization (ELDI) source. This is the first demonstration of the use of building blocks to construct and test the TLC-MS interfacing system. With the advantages of being readily available, cheap, reusable, and extremely easy to modify without consuming any material or reagent, the use of building blocks to develop the TLC-AMS interface is undoubtedly a green methodology. The TLC plate delivery system consists of a storage box, plate dealing component, conveyer, light sensor, and plate collecting box. During a TLC-AMS analysis, the TLC plate was sent to the conveyer from a stack of TLC plates placed in the storage box. As the TLC plate passed through the ELDI source, the chemical compounds separated on the plate would be desorbed by laser desorption and subsequently postionized by electrospray ionization. The samples, including a mixture of synthetic dyes and extracts of pharmaceutical drugs, were analyzed to demonstrate the capability of this TLC-ELDI/MS system for high-throughput analysis.

  16. Application of 2-Trichloromethylbenzimidazole in Analytical Chemistry: A Highly Selective Chromogenic Reagent for Thin-Layer Chromatography and Some Other Analytical Uses

    Directory of Open Access Journals (Sweden)

    Leszek Konopski

    2012-01-01

    Full Text Available 2-Trichloromethylbenzimidazole (TCMB was used as a chromogenic reagent in organic or inorganic analysis, mainly in thin-layer chromatography (TLC. In reactions of TCMB with some heteroaromatic nitrogen containing compounds, such as azines, azoles and benzazoles, a formation of high colored products occurred. For azines, the chromogenic reaction was highly regioselective, since the both adjacent α-positions versus the nitrogen atom(s must not be substituted. A TLC method of detection was developed. Thirty azines, azoles, and benzazoles were detected at the detection limit 10 ng to 1 μg. This method was also applied for detection of heteroaromatic pesticides, and the attempts to construct active and passive dosimeters for nicotine were made. In a prechromatographic reaction of aromatic o-diamines with methyl trichloroacetimidate, TCMB or its derivatives were formed in situ. Followed by TLC and visualization in pyridine vapors, this procedure was applied for detection of o-phenylenediamine derivatives. The reaction product of TCMB and pyridine (LI Complex was identified and fully characterized. Two different reaction mechanisms: with electron deficient basic heteroaromatic compounds, like pyridine, and with more acidic compounds, for example, pyrrole, were discussed. In aqueous solutions, the LI Complex may be also used as a new indicator for complexometric, adsorption and acid-base titration of inorganic compounds.

  17. A separable surface-enhanced Raman scattering substrate modified with MIL-101 for detection of overlapping and invisible compounds after thin-layer chromatography development.

    Science.gov (United States)

    Zhang, Bin Bin; Shi, Yi; Chen, Hui; Zhu, Qing Xia; Lu, Feng; Li, Ying Wei

    2018-01-02

    By coupling surface-enhanced Raman spectroscopy (SERS) with thin-layer chromatography (TLC), a powerful method for detecting complex samples was successfully developed. However, in the TLC-SERS method, metal nanoparticles serving as the SERS-active substrate are likely to disturb the detection of target compounds, particularly in overlapping compounds after TLC development. In addition, the SERS detection of compounds that are invisible under both visible light and UV 254/365 after TLC development is still a significant challenge. In this study, we demonstrated a facile strategy to fabricate a TLC plate with metal-organic framework-modified gold nanoparticles as a separable SERS substrate, on which all separated components, including overlapping and invisible compounds, could be detected by a point-by-point SERS scan along the developing direction. Rhodamine 6G (R6G) was used as a probe to evaluate the performance of the substrate. The results indicated that the substrate provided good sensitivity and reproducibility, and optimal SERS signals could be collected in 5 s. Furthermore, this new substrate exhibited a long shelf life. Thus, our method has great potential for the sensitive and rapid detection of overlapping and invisible compounds in complex samples after TLC development. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.

  18. Detection of tobacco-related biomarkers in urine samples by surface-enhanced Raman spectroscopy coupled with thin-layer chromatography.

    Science.gov (United States)

    Huang, Rongfu; Han, Sungyub; Li, Xiao Sheryl

    2013-08-01

    The nicotine metabolites, cotinine and trans-3'-hydroxycotinine (3HC) are considered as superior biomarkers for identifying tobacco exposure. More importantly, the ratio of 3HC to cotinine is a good indicator to phenotype individuals for cytochrome P450 2A6 activity and to individualize pharmacotherapy for tobacco addiction. In this paper, a simple, robust and novel method based on surface-enhanced Raman spectroscopy coupled with thin-layer chromatography (TLC) was developed to directly quantify the biomarkers in human urine samples. This is the first time surface-enhanced Raman spectroscopy (SERS) was used to detect cotinine and 3HC in urine samples. The linear dynamic range for the detection of cotinine is from 40 nM to 8 μM while that of 3HC is from 1 μM to 15 μM. The detection limits are 10 nM and 0.2 μM for cotinine and 3HC, respectively. The proposed method was further validated by quantifying the concentration of both cotinine and 3HC in smokers' urine samples. This TLC-SERS method allows the direct detection of cotinine in the urine samples of both active and passive smokers and the detection of 3HC in smokers.

  19. Rapid Biochemical Mixture Screening by Three-Dimensional Patterned Multifunctional Substrate with Ultra-Thin Layer Chromatography (UTLC) and Surface Enhanced Raman Scattering (SERS).

    Science.gov (United States)

    Lee, Bi-Shen; Lin, Pi-Chen; Lin, Ding-Zheng; Yen, Ta-Jen

    2018-01-11

    We present a three-dimensional patterned (3DP) multifunctional substrate with the functions of ultra-thin layer chromatography (UTLC) and surface enhanced Raman scattering (SERS), which simultaneously enables mixture separation, target localization and label-free detection. This multifunctional substrate is comprised of a 3DP silicon nanowires array (3DP-SiNWA), decorated with silver nano-dendrites (AgNDs) atop. The 3DP-SiNWA is fabricated by a facile photolithographic process and low-cost metal assisted chemical etching (MaCE) process. Then, the AgNDs are decorated onto 3DP-SiNWA by a wet chemical reduction process, obtaining 3DP-AgNDs@SiNWA multifunctional substrates. With various patterns designed on the substrates, the signal intensity could be maximized by the excellent confinement and concentrated effects of patterns. By using this 3DP-AgNDs@SiNWA substrate to scrutinize the mixture of two visible dyes, the individual target could be recognized and further boosted the Raman signal of target 15.42 times comparing to the un-patterned AgNDs@SiNWA substrate. Therefore, such a three-dimensional patterned multifunctional substrate empowers rapid mixture screening, and can be readily employed in practical applications for biochemical assays, food safety and other fields.

  20. Detection of structurally similar adulterants in botanical dietary supplements by thin-layer chromatography and surface enhanced Raman spectroscopy combined with two-dimensional correlation spectroscopy.

    Science.gov (United States)

    Li, Hao; Zhu, Qing xia; Chwee, Tsz sian; Wu, Lin; Chai, Yi feng; Lu, Feng; Yuan, Yong fang

    2015-07-09

    Thin-layer chromatography (TLC) coupled with surface enhanced Raman spectroscopy (SERS) has been widely used for the study of various complex systems, especially for the detection of adulterants in botanical dietary supplements (BDS). However, this method is not sufficient to distinguish structurally similar adulterants in BDS since the analogs have highly similar chromatographic and/or spectroscopic behaviors. Taking into account the fact that higher cost and more time will be required for comprehensive chromatographic separation, more efforts with respect to spectroscopy are now focused on analyzing the overlapped SERS peaks. In this paper, the combination of a TLC-SERS method with two-dimensional correlation spectroscopy (2DCOS), with duration of exposure to laser as the perturbation, is applied to solve this problem. Besides the usual advantages of the TLC-SERS method, such as its simplicity, rapidness, and sensitivity, more advantages are presented here, such as enhanced selectivity and good reproducibility, which are obtained by 2DCOS. Two chemicals with similar structures are successfully differentiated from the complex BDS matrices. The study provides a more accurate qualitative screening method for detection of BDS with adulterants, and offers a new universal approach for the analysis of highly overlapped SERS peaks. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Construction of a supercritical fluid extraction (SFE equipment: validation using annatto and fennel and extract analysis by thin layer chromatography coupled to image

    Directory of Open Access Journals (Sweden)

    Júlio Cezar Flores JOHNER

    2016-01-01

    Full Text Available Abstract The present work describes setting up a laboratory unit for supercritical fluid extraction. In addition to its construction, a survey of cost was done to compare the cost of the homemade unit with that of commercial units. The equipment was validated using an extraction of annatto seeds’ oil, and the extraction and fractionation of fennel oil were used to validate the two separators; for both systems, the solvent was carbon dioxide. The chemical profiles of annatto and fennel extracts were assessed using thin layer chromatography; the images of the chromatographic plates were processed using the free ImageJ software. The cost survey showed that the homemade equipment has a very low cost (~US$ 16,000 compared to commercial equipment. The extraction curves of annatto were similar to those obtained in the literature (yield of 3.8% oil. The separators were validated, producing both a 2.5% fraction of fennel seed extract rich in essential oils and another extract fraction composed mainly of oleoresins. The ImageJ software proved to be a low-cost tool for obtaining an initial evaluation of the chemical profile of the extracts.

  2. High-performance Thin-layer Chromatographic-densitometric Quantification and Recovery of Bioactive Compounds for Identification of Elite Chemotypes of Gloriosa superba L. Collected from Sikkim Himalayas (India).

    Science.gov (United States)

    Misra, Ankita; Shukla, Pushpendra Kumar; Kumar, Bhanu; Chand, Jai; Kushwaha, Poonam; Khalid, Md; Singh Rawat, Ajay Kumar; Srivastava, Sharad

    2017-10-01

    Gloriosa superba L. (Colchicaceae) is used as adjuvant therapy in gout for its potential antimitotic activity due to high colchicine(s) alkaloids. This study aimed to develop an easy, cheap, precise, and accurate high-performance thin-layer chromatographic (HPTLC) validated method for simultaneous quantification of bioactive alkaloids (colchicine and gloriosine) in G. superba L. and to identify its elite chemotype(s) from Sikkim Himalayas (India). The HPTLC chromatographic method was developed using mobile phase of chloroform: acetone: diethyl amine (5:4:1) at λ max of 350 nm. Five germplasms were collected from targeted region, and on morpho-anatomical inspection, no significant variation was observed among them. Quantification data reveal that content of colchicine ( R f : 0.72) and gloriosine ( R f : 0.61) varies from 0.035%-0.150% to 0.006%-0.032% (dry wt. basis). Linearity of method was obtained in the concentration range of 100-400 ng/spot of marker(s), exhibiting regression coefficient of 0.9987 (colchicine) and 0.9983 (gloriosine) with optimum recovery of 97.79 ± 3.86 and 100.023% ± 0.01%, respectively. Limit of detection and limit of quantification were analyzed, respectively, as 6.245, 18.926 and 8.024, 24.316 (ng). Two germplasms, namely NBG-27 and NBG-26, were found to be elite chemotype of both the markers. The developed method is validated in terms of accuracy, recovery, and precision studies as per the ICH guidelines (2005) and can be adopted for the simultaneous quantification of colchicine and gloriosine in phytopharmaceuticals. In addition, this study is relevant to explore the chemotypic variability in metabolite content for commercial and medicinal purposes. An easy, cheap, precise, and accurate high performance thin layer chromatographic (HPTLC) validated method for simultaneous quantification of bioactive alkaloids (colchicine and gloriosine) in G. superba L.Five germplasms were collected from targeted region, and on morpho anatomical

  3. Slantingly cross loading sample system enables simultaneous performance of separation and mixture to detect molecular interactions on thin-layer chromatography.

    Science.gov (United States)

    Shimizu-Yumoto, Hiroko; Hayashi, Nobuyuki; Ichimura, Kazuo; Nakayama, Masayoshi

    2012-07-06

    Anthocyanins are major flower pigments that can be affected by copigments, colorless compounds that can modify anthocyanin coloration to more intense and bluer. Thin-layer chromatography (TLC) is an available technique to separate and analyze anthocyanins and copigments. To easily and comprehensively detect copigments, we added function of mixture of compounds to TLC; by slantingly cross loading samples on TLC, compounds are symmetrically developed at various angle lines from the upper origin to individual R(f) values and cross each other in an orderly fashion, where mixture is simultaneously performed with separation. Occurrence of copigments can be detected as a coloration change on the developed line of anthocyanin. Pink sweet pea (Lathyrus odoratus L.) petals were analyzed by the cross-TLC and a more intense spot and a paler spot on the anthocyanin line were detected. As each spot overlapped with an ultraviolet absorbance line, each of these ultraviolet absorption compounds was purified and identified as kaempferol 3-rhamnoside and 2-cyanoethyl-isoxazolin-5-one, respectively. Whereas kaempferol 3-rhamnoside is a flavonoid and had a general copigment effect of more intense and bluer coloration change, 2-cyanoethyl-isoxazolin-5-one is a compound whose structure is outside of conventional categories of copigments and had a novel effect to change anthocyanin coloration paler while maintaining color tone. We determined that the search for copigments should be carried out without pre-existing prediction of structures and effects. We have shown that slantingly cross loading samples system on plate-type chromatography is an effective technique for such comprehensive analysis of molecular interaction. Crown Copyright © 2012. Published by Elsevier B.V. All rights reserved.

  4. High Performance Liquid Chromatography Experiments to Undergraduate Laboratories

    Science.gov (United States)

    Kissinger, Peter T.; And Others

    1977-01-01

    Reviews the principles of liquid chromatography with electrochemical detection (LCEC), an analytical technique that incorporates the advantages of both liquids chromatography and electrochemistry. Also suggests laboratory experiments using this technique. (MLH)

  5. Separation of anionic oligosaccharides by high-performance liquid chromatography

    International Nuclear Information System (INIS)

    Green, E.D.; Baenziger, J.U.

    1986-01-01

    The authors have developed methods for rapid fractionation of anionic oligosaccharides containing sulfate and/or sialic acid moieties by high-performance liquid chromatography (HPLC). Ion-exchange HPLC on amine-bearing columns (Micropak AX-10 and AX-5) at pH 4.0 is utilized to separate anionic oligosaccharides bearing zero, one, two, three, or four charges, independent of the identity of the anionic moieties (sulfate and/or sialic acid). Ion-exchange HPLC at pH 1.7 allows separation of neutral, mono-, di-, and tetrasialylated, monosulfated, and disulfated oligosaccharides. Oligosaccharides containing three sialic acid residues and those bearing one each of sulfate and sialic acid, however, coelute at pH 1.7. Since the latter two oligosaccharide species separate at pH 4.0, analysis at pH 4.0 followed by analysis at pH 1.7 can be utilized to completely fractionate complex mixtures of sulfated and sialylated oligosaccharides. Ion-suppression amine adsorption HPLC has previously been shown to separate anionic oligosaccharides on the basis of net carbohydrate content (size). In this study they demonstrate the utility of ion-suppression amine adsorption HPLC for resolving sialylated oligosaccharide isomers which differ only in the linkages of sialic acid residues (α2,3 vs α2,6) and/or location of α2,3- and α2,6-linked sialic acid moieties on the peripheral branches of oligosaccharides. These two methods can be used in tandem to separate oligosaccharides, both analytically and preparatively, based on their number, types, and linkages of anionic moieties

  6. High-performance thin-layer chromatographic methods in the evaluation of the antioxidant and anti-hyperglycemic activity of Myrmecodia platytyrea as a promising opportunity in diabetes treatment.

    Science.gov (United States)

    Agatonovic-Kustrin, S; Morton, D W; Adam, A; Mizaton, H H; Zakaria, H

    2017-12-29

    The steady increase of diabetes is becoming a major burden on health care systems. As diabetic complications arise from oxidative stress, an antioxidant therapy along with anti-diabetic drugs is recommended. Myrmecodia or ant plant is highly valued as a traditional medicine in West Papua. It is used as an alternative treatment for diabetes, as the substances produced by ants can reduce blood sugar levels. The aim of this study was to develop and establish high-performance thin-layer chromatographic (HPTLC)-bioautographic methods to measure the antioxidant and hypoglycemic effects in different extracts from Myrmecodia platytyrea and to compare them with sterol content. Antioxidant activity in methanol, ethanol, dichloromethane (DCM) and ethyl acetate (EA) extracts were measured with a direct HPTLC-2,2-diphenyl-1-picrylhydrazyl free radical (DPPH) assay, while hypoglycemic effects were assessed using a newly developed α-amylase inhibitory activity assay. Stigmasterol is observed, after derivatization with anisaldehyde, as purple colored zones under visible light at hRF values of 0.66. The highest antioxidant activity was observed in the ethanol extract which is rich in polyphenols and flavonoids, while the DCM extract did not show antioxidant activity, but had significant α-amylase inhibitory activity. The highest α-amylase inhibitory activity was observed in the EA and DCM extracts and was related to their stigmasterol content. Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.

  7. [High-performance liquid-liquid chromatography in beverage analysis].

    Science.gov (United States)

    Bricout, J; Koziet, Y; de Carpentrie, B

    1978-01-01

    Liquid liquid chromatography was performed with columns packed with stationary phases chemically bonded to silica microparticules. These columns show a high efficiency and are used very easily. Flavouring compounds like aromatic aldehydes which have a low volatility were analyzed in brandy using a polar phase alkylnitrile. Sapid substances like amarogentin in Gentiana lutea or glyryrrhizin in Glycyrrhiza glabra were determined by reversed phase chromatography. Finally ionizable substances like synthetic dyes can be analyzed by paired ion chromatography witha non polar stationary phase.

  8. A thin-layer chromatography method for the identification of three different olibanum resins (Boswellia serrata, Boswellia papyrifera and Boswellia carterii, respectively, Boswellia sacra).

    Science.gov (United States)

    Paul, Michael; Brüning, Gerit; Bergmann, Jochen; Jauch, Johann

    2012-01-01

    Resins of the genus Boswellia are currently an interesting topic for pharmaceutical research since several pharmacological activities (e.g. anti-inflammatory, anti-microbial, anti-tumour) are reported for extracts and compounds isolated from them. Unambiguous identification of these resins, by simple and convenient analytical methods, has so far not clearly been verified. For differentiation and identification of three important Boswellia species (Boswellia serrata Roxb., Boswellia papyrifera Hochst. and Boswellia carterii Birdw., respectively Boswellia sacra Flueck.), possible even for minimally equipped laboratories, a thin-layer chromatography (TLC) method was developed, allowing unambiguous identification of the three species. Crude resin samples (commercial samples and a voucher specimen) were extracted with methanol or diethyl ether and subjected to TLC analysis (normal phase). A pentane and diethyl ether (2:1) with 1% acetic acid eluent was used. Chromatograms were analysed by UV detection (254 nm) and dyeing with anisaldehyde dyeing reagent. Significant spots were isolated and structures were assigned (mass spectrometry; nuclear magnetic resonance spectroscopy). Incensole and incensole acetate are specific biomarkers for Boswellia papyrifera. Boswellia carterii/Boswellia sacra reveal ß-caryophyllene oxide as a significant marker compound. Boswellia serrata shows neither incensole acetate nor ß-caryophyllene oxide spots, but can be identified by a strong serratol and a sharp 3-oxo-8,24-dien-tirucallic acid spot. The TLC method developed allows unambiguous identification of three different olibanum samples (Boswellia papyrifera, Boswellia serrata, Boswellia carterii/Boswellia sacra). Evidence on the specific biosynthesis routes of these Boswellia species is reported. Copyright © 2011 John Wiley & Sons, Ltd.

  9. Thin layer chromatography coupled with surface-enhanced Raman scattering as a facile method for on-site quantitative monitoring of chemical reactions.

    Science.gov (United States)

    Zhang, Zong-Mian; Liu, Jing-Fu; Liu, Rui; Sun, Jie-Fang; Wei, Guo-Hua

    2014-08-05

    By coupling surface-enhanced Raman spectroscopy (SERS) with thin layer chromatography (TLC), a facile and powerful method was developed for on-site monitoring the process of chemical reactions. Samples were preseparated on a TLC plate following a common TLC procedure, and then determined by SERS after fabricating a large-area, uniform SERS substrate on the TLC plate by spraying gold nanoparticles (AuNPs). Reproducible and strong SERS signals were obtained with substrates prepared by spraying 42-nm AuNPs at a density of 5.54 × 10(10) N/cm(2) on the TLC plate. The capacity of this TLC-SERS method was evaluated by monitoring a typical Suzuki coupling reaction of phenylboronic acid and 2-bromopyridine as a model. Results showed that this proposed method is able to identify reaction product that is invisible to the naked eye, and distinguish the reactant 2-bromopyridine and product 2-phenylpyridine, which showed almost the same retention factors (R(f)). Under the optimized conditions, the peak area of the characteristic Raman band (755 cm(-1)) of the product 2-phenylpyridine showed a good linear correlation with concentration in the range of 2-200 mg/L (R(2) = 0.9741), the estimated detection limit (1 mg/L 2-phenylpyridine) is much lower than the concentration of the chemicals in the common organic synthesis reaction system, and the product yield determined by the proposed TLC-SERS method agreed very well with that by UPLC-MS/MS. In addition, a new byproduct in the reaction system was found and identified through continuous Raman detection from the point of sample to the solvent front. This facile TLC-SERS method is quick, easy to handle, low-cost, sensitive, and can be exploited in on-site monitoring the processes of chemical reactions, as well as environmental and biological processes.

  10. Determination of Sudan I in paprika powder by molecularly imprinted polymers-thin layer chromatography-surface enhanced Raman spectroscopic biosensor.

    Science.gov (United States)

    Gao, Fang; Hu, Yaxi; Chen, Da; Li-Chan, Eunice C Y; Grant, Edward; Lu, Xiaonan

    2015-10-01

    Sudan I is a carcinogenic and mutagenic azo-compound that has been utilized as a common adulterant in spice and spice blends to impart a desirable red color to foods. A novel biosensor combining molecularly imprinted polymers (MIPs), thin layer chromatography (TLC) and surface enhanced Raman spectroscopy (SERS) could determine Sudan I levels in paprika powder to 1 ppm (or 2 ng/spot). Sudan I spiked paprika extracts (spiking levels: 0, 1, 5, 10, 40, 70 and 100 ppm) were prepared. Sudan I imprinted polymers were synthesized by employing the interaction between Sudan I (template) and methacrylic acid (functional monomer), followed by washing to remove Sudan I leaving the Sudan I-binding sites exposed. MIPs were used as a stationary phase for TLC and could selectively retain Sudan I at the original spot with little interference. A gold colloid SERS substrate could enhance Raman intensity for Sudan I in this MIP-TLC system. Principal component analysis plot and partial least squares regression (R(2)=0.978) models were constructed and a linear regression model (R(2)=0.983) correlated spiking levels (5, 10, 40, 70 and 100 ppm) with the peak intensities (721 cm(-1)) of Sudan I SERS spectra. Both separation (30-40s) and detection (1s or 0.1s) were extremely fast by using both commercial bench-top and custom made portable Raman spectrometers. This biosensor can be applied as a rapid, low-cost and reliable tool for screening Sudan I adulteration in foods. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Normal Raman and surface enhanced Raman spectroscopic experiments with thin layer chromatography spots of essential amino acids using different laser excitation sources

    Science.gov (United States)

    István, Krisztina; Keresztury, Gábor; Szép, Andrea

    2003-06-01

    A comparative study of the feasibility and efficiency of Raman spectroscopic detection of thin layer chromatography (TLC) spots of some weak Raman scatterers (essential amino acids, namely, glycine and L-forms of alanine, serine, valine, proline, hydroxyproline, and phenylalanine) was carried out using four different visible and near-infrared (NIR) laser radiations with wavelengths of 532, 633, 785, and 1064 nm. Three types of commercial TLC plates were tested and the possibility of inducing surface enhanced Raman scattering (SERS) by means of Ag-sol was also investigated. The spectra obtained from spotted analytes adsorbed on TLC plates were of very different quality strongly depending on the excitation wavelength, the wetness of the samples, and the compounds examined. The best results were obtained with the simple silica TLC plate, and it has been established that the longest wavelength (lowest energy) NIR excitation of a Nd:YAG laser is definitely more suitable for generating normal Raman scattering of analyte spots than any of the visible radiations. Concerning SERS with application of Ag-sol to the TLC spots, 1-3 orders of magnitude enhancement was observed with wet samples, the greatest with the 532 nm radiation and gradually smaller with the longer wavelength excitations. It is shown, however, that due to severe adsorption-induced spectral distortions and increased sensitivity to microscopic inhomogeneity of the sample, none of the SERS spectra obtained with the dispersive Raman microscope operating in the visible region were superior to the best NIR normal FT-Raman spectra, as far as sample identification is concerned.

  12. Facile on-site detection of substituted aromatic pollutants in water using thin layer chromatography combined with surface-enhanced Raman spectroscopy.

    Science.gov (United States)

    Li, Dawei; Qu, Lulu; Zhai, Wenlei; Xue, Jinqun; Fossey, John S; Long, Yitao

    2011-05-01

    A novel facile method for on-site detection of substituted aromatic pollutants in water using thin layer chromatography (TLC) combined with surface-enhanced Raman spectroscopy (SERS) was explored. Various substituted aromatics in polluted water were separated by a convenient TLC protocol and then detected using a portable Raman spectrometer with the prepared silver colloids serving as SERS-active substrates. The effects of operating conditions on detection efficacy were evaluated, and the application of TLC-SERS to on-site detection of artificial and real-life samples of aromatics/polluted water was systematically investigated. It was shown that commercially available Si 60-F(254) TLC plates were suitable for separation and displayed low SERS background and good separation efficiency, 2 mM silver colloids, 20 mM NaCl (working as aggregating agent), 40 mW laser power, and 50 s intergration time were appropriate for the detection regime. Furthermore, qualitative and quantitative detection of most of substituted aromatic pollutants was found to be readily accomplished using the developed TLC-SERS technique, which compared well with GC-MS in terms of identification ability and detection accuracy, and a limit of detection (LOD) less than 0.2 ppm (even at ppb level for some analytes) could be achieved under optimal conditions. The results reveal that the presented convenient method could be used for the effective separation and detection of the substituted aromatic pollutants of water on site, thus reducing possible influences of sample transportation and contamination while shortening the overall analysis time for emergency and routine monitoring of the substituted aromatics/polluted water.

  13. Toward rapid analysis, forecast and discovery of bioactive compounds from herbs by jointly using thin layer chromatography and ratiometric surface-enhanced Raman spectroscopy technique.

    Science.gov (United States)

    Gu, Xiaoling; Jin, Yang; Dong, Fang; Cai, Yueqing; You, Zhengyi; You, Junhui; Zhang, Liying; Du, Shuhu

    2018-05-10

    Conventional isolation and identification of active compounds from herbs have been extensively reported by using various chromatographic and spectroscopic techniques. However, how to quickly discover new bioactive ingredients from natural sources still remains a challenging task due to the interference of their similar structures or matrices. Here, we present a grand approach for rapid analysis, forecast and discovery of bioactive compounds from herbs based on a hyphenated strategy of thin layer chromatography and ratiometric surface-enhanced Raman spectroscopy. The performance of the hyphenated strategy is first evaluated by analyzing four protoberberine alkaloids, berberine (BER), coptisine (COP), palmatine (PAT) and jatrorrhizine (JAT), from a typical herb Coptidis Rhizoma as an example. It has been demonstrated that this coupling method can identify the four compounds by characteristic peaks at 728, 708, 736 and 732 cm -1 , and especially discriminate BER and COP (with similar migration distances) by ratiometric Raman intensity (I 708 /I 728 ). The corresponding limits of detection are 0.1, 0.05, 0.1 and 0.5 μM, respectively, which are about 1-2 orders of magnitude lower than those of direct observation method under 254 nm UV lamp. Based on these findings, the proposed method further guides forecast and discovery of unknown compounds from traditional Chinese herb Typhonii Rhizoma. Results infer that two trace alkaloids (BER and COP) from the n-butanol extract of Typhonii Rhizoma are found for the first time. Moreover, in vitro experiments manifest that BER can effectively decrease the viability of human glioma U87 cells by inducing cell cycle arrest in a concentration-dependent manner. Copyright © 2018 Elsevier B.V. All rights reserved.

  14. Determination of trimethyllead reference material using high performance liquid chromatography-inductively coupled plasma mass spectrometry

    International Nuclear Information System (INIS)

    Lu Hai; Wei Chao; Wang Jun; Chao Jingbo; Zhou Tao; Chen Dazhou

    2005-01-01

    A high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICPMS) was combined, and the chromatography conditions were optimized. The stability and homogeneity of a trimethyllead reference material were determined using this method. (authors)

  15. Microchip-based monolithic column for high performance liquid chromatography

    Data.gov (United States)

    National Aeronautics and Space Administration — We have developed microchip based monolithic columns that can be used for liquid chromatography of small organic molecules, as well as, macromolecules such as...

  16. High Performance Liquid Chromatography of Vitamin A: A Quantitative Determination.

    Science.gov (United States)

    Bohman, Ove; And Others

    1982-01-01

    Experimental procedures are provided for the quantitative determination of Vitamin A (retinol) in food products by analytical liquid chromatography. Standard addition and calibration curve extraction methods are outlined. (SK)

  17. Stable, Microfabricated Thin Layer Chromatography Plates without Volume Distortion on Patterned, Carbon and Al2O3-Primed Carbon Nanotube Forests

    Energy Technology Data Exchange (ETDEWEB)

    Jensen, David S.; Kanyal, Supriya S.; Gupta, Vipul; Vail, Michael A.; Dadson, Andrew; Engelhard, Mark H.; Vanfleet, Richard; Davis, Robert C.; Linford, Matthew R.

    2012-09-28

    In a recent report (Song, J.; et al., Advanced Functional Materials 2011, 21, 1132-1139) some of us described the fabrication of thin layer chromatography (TLC) plates from patterned carbon nanotube (CNT) forests, which were directly infiltrated/coated with silicon by low pressure chemical vapor deposition (LPCVD) of silicon using SiH4. Following infiltration, the nanotubes were removed from the assemblies and the silicon simultaneously converted to SiO2 in a high temperature oxidation step. However, while straightforward, this process had some shortcomings, not the least of which was some distortion of the lithographically patterned features during the volume expansion that accompanied oxidation. Herein we overcome theis issue and also take substantial steps forward in the microfabrication of TLC plates by showing: (i) A new method for creating an adhesion promotion layer on CNT forests by depositing a few nanometers of carbon followed by atomic layer deposition (ALD) of Al2O3. This method for appears to be new, and X-ray photoelectron spectroscopy confirms the expected presence of oxygen after carbon deposition. ALD of Al2O3 alone and in combination with the carbon on patterned CNT forests was also explored as an adhesion promotion layer for CNT forest infiltration. (ii) Rapid, conformal deposition of an inorganic material that does not require subsequent oxidation: fast pseudo-ALD growth of SiO2 via alumina catalyzed deposition of tris(tert-butoxy)silanol onto the carbon/Al2O3-primed CNT forests. (iii) Faithful reproduction of the features in the masks used to microfabricate the TLC plates (M-TLC) this advance springs from the previous two points. (iv) A bonded (amino) phase on a CNT-templated microfabricated TLC plate. (v) Fast, highly efficient (125,000 - 225,000 N/m) separations of fluorescent dyes on M-TLC plates. (vi) Extensive characterization of our new materials by TEM, SEM, EDAX, DRIFT, and XPS. (vii) A substantially lower process temperature for the

  18. The importance of thin layer chromatography and UV microspectrophotometry in the analysis of reactive dyes released from wool and cotton fibers.

    Science.gov (United States)

    Wiggins, Kenneth G; Holness, Julie-Ann; March, Bridget M

    2005-03-01

    Samples of reactively-dyed wool and cotton were obtained from a range of dye manufacturers, dye distributors and the Forensic Science Service (FSS) Fibre Data Collection. The wool fibers were red in color and had previously been compared using comparison microscopy (CM), visible range microspectrophotometry (VS) and thin layer chromatography (TLC). The cotton fibers were blue and black in color and had not been previously compared. Red, blue and black fibers were chosen because they are often encountered in casework. The usage of reactive dyes to color fibers has increased over the last 10-15 years and these are often seen in casework. Before techniques were available that allowed reactively-dyed fibers to be compared using TLC only CM and microspectrophotometry were routinely carried out. Many laboratories, who had a microspectrophotometer, only had a visible range instrument. It was therefore important to see which techniques provide additional information, that gives greater individuality to fibers, to that obtained from CM. The color was released from the wool and cotton fibres using alkaline hydrolysis and a cellulase enzyme respectively. Many of the red wool samples were differentiated from each other using CM. More differentiation was found using VS and even more when ultraviolet range microspectrophotometry (UV) or TLC was used. Two samples could only be differentiated using TLC because CM, VS and UV failed to separate them. The black cotton samples were predominately differentiated using CM but VS allowed for further differentiation. With the samples used in this project UV and TLC failed to separate the samples further. The blue cotton samples benefited from the use of CM, VS and either UV or TLC to reduce the number of matching pairs. All techniques aided differentiation although with this set TLC and UV proved to be complementary techniques. Results demonstrate that TLC and UV both yield important information over and above that obtained from CM and VS

  19. Separation of paralytic shellfish poisoning toxins on Chromarods-SIII by thin-layer chromatography with the Iatroscan (mark 5) and flame thermionic detection.

    Science.gov (United States)

    Indrasena, W M; Ackman, R G; Gill, T A

    1999-09-10

    Thin-layer chromatography (TLC) on Chromarods-SIII with the Iatroscan (Mark-5) and a flame thermionic detector (FTID) was used to develop a rapid method for the detection of paralytic shellfish poisoning (PSP) toxins. The effect of variation in hydrogen (H2) flow, air flow, scan time and detector current on the FTID peak response for both phosphatidylcholine (PC) and PSP were studied in order to define optimum detection conditions. A combination of hydrogen and air flow-rates of 50 ml/min and 1.5-2.0 l/min respectively, along with a scan time of 40 s/rod and detector current of 3.0 A (ampere) or above were found to yield the best results for the detection of PSP compounds. Increasing the detector current level to as high as 3.3 A gave about 130 times more FTID response than did flame ionization detection (FID), for PSP components. Quantities of standards as small as 1 ng neosaxitoxin (NEO), 5 ng saxitoxin (STX), 5 ng B1-toxins (B1), 2 ng gonyautoxin (GTX) 2/3, 6 ng GTX 1/4 and 6 ng C-toxins (C1/C2) could be detected with the FTID. The method detection limits for toxic shellfish tissues using the FTID were 0.4, 2.1, 0.8 and 2.5 micrograms per g tissue for GTX 2/3, STX, NEO and C toxins, respectively. The FTID response increased with increasing detector current and with increasing the scan time. Increasing hydrogen and air flow-rates resulted in decreasing sensitivity within defined limits. Numerous solvent systems were tested, and, solvent consisting of chloroform: methanol-water-acetic acid (30:50:8:2) could separate C toxins from GTX, which eluted ahead of NEO and STX. Accordingly, TLC/FTID with the Iatroscan (Mark-5) seems to be a promising, relatively inexpensive and rapid method of screening plant and animal tissues for PSP toxins.

  20. Analysis of short-chain acids from anaerobic bacteria by high-performance liquid chromatography.

    OpenAIRE

    Guerrant, G O; Lambert, M A; Moss, C W

    1982-01-01

    A standard mixture of 25 short-chain fatty acids was resolved by high-performance liquid chromatography, using an Aminex HPX-87 column. The acids produced in culture media by anaerobic bacteria were analyzed by high-performance liquid chromatography after extraction with ether and reextraction into a small volume of 0.1 N NaOH. The presence of fumaric acid in culture extracts of Peptostreptococcus anaerobius was confirmed by gas chromatography-mass spectrometry analysis of the trapped eluent ...

  1. Resolution of RNA using high-performance liquid chromatography

    NARCIS (Netherlands)

    Mclaughlin, L.W.; Bischoff, Rainer

    1987-01-01

    High-performance liquid chromatographic techniques can be very effective for the resolution and isolation of nucleic acids. The characteristic ionic (phosphodiesters) and hydrophobic (nucleobases) properties of RNAs can be exploited for their separation. In this respect anion-exchange and

  2. Buffer-Free High Performance Liquid Chromatography Method for ...

    African Journals Online (AJOL)

    Purpose: To develop and validate a simple, economical and reproducible high performance liquid chromatographic (HPLC) method for the determination of theophylline in pharmaceutical dosage forms. Method: Caffeine was used as the internal standard and reversed phase C-18 column was used to elute the drug and ...

  3. High-performance liquid chromatography of human glycoprotein hormones.

    Science.gov (United States)

    Chlenov, M A; Kandyba, E I; Nagornaya, L V; Orlova, I L; Volgin, Y V

    1993-02-12

    The chromatographic behavior of the glycoprotein hormones from human pituitary glands and of placental origin [thyroid-stimulating hormone, luteinizing hormone and chorionic gonadotropin (CG)] was studied. It was shown that hydrophobic interaction chromatography on a microparticulate packing and anion-exchange HPLC can be applied for the purification of these hormones. Reversed-phase HPLC on wide-pore C4-bonded silica at neutral pH can be applied for the determination of the above hormones and for the isolation of pure CG and its subunits.

  4. Microcystin Detection Characteristics of Fluorescence Immunochromatography and High Performance Liquid Chromatography

    International Nuclear Information System (INIS)

    Pyo, Dong Jin; Park, Geun Young; Choi, Jong Chon; Oh, Chang Suk

    2005-01-01

    Different detection characteristics of fluorescence immunochromatography method and high performance liquid chromatography (HPLC) method for the analysis of cyanobacterial toxins were studied. In particular, low and high limits of detection, detection time and reproducibility and detectable microcystin species were compared when fluorescence immunochromatography method and high performance liquid chromatography method were applied for the detection of microcystin (MC), a cyclic peptide toxin of the freshwater cyanobacterium Microcystis aeruginosa. A Fluorescence immunochromatography assay system has the unique advantages of short detection time and low detection limit, and high performance liquid chromatography detection method has the strong advantage of individual quantifications of several species of microcystins

  5. Determination of pyrazinamide in human by high performance liquid chromatography.

    Directory of Open Access Journals (Sweden)

    Revankar S

    1994-01-01

    Full Text Available A facile and sensitive high performance liquid chromatographic (HPLC technique has been developed for the determination pyrazinamide (PZA in human plasma. Nicotinamide(NIA is used as internal standard(IS. Plasma is deproteinized with 0.7 M perchloric acid; clear supernatant is neutralized with 1M NaOH and injected onto HPLC. The separation of pyrazinamide and the internal standard is carried out on a Supelco LC-18 (DB column with a basic mobile phase. Pyrazinoic acid, the major metabolite, other anti-tuberculous drugs and endogenous components do not interfere with measurement of pyrazinamide. The limit of detection of pyrazinamide with this method is 0.2 mg/0.2 ml plasma (CV 8.2%.

  6. Separation and estimation of lanthanides using high performance liquid chromatography

    International Nuclear Information System (INIS)

    Datta, Arpita; Sivaraman, N.; Vasudeva Rao, P.R.

    2012-01-01

    The separation efficiency of individual lanthanides depends on the stability constant of the metal-ligand complex. Therefore, stability constant data of lanthanide complexes is important in the development of high performance separation procedures. The dynamic ion exchange HPLC technique was employed at our laboratory to estimate the stability constant of lanthanides with various complexing agents. In these studies, the retention times as well as capacity factors of lanthanides and some actinides were measured as a function of CSA, complexing agent concentrations and mobile phase pH. From these studies, a correlation has been established between capacity factor of a metal ion, concentrations of ion-pairing reagent and complexing agent with the stability constant of lanthanide complex

  7. Thin layer activation

    International Nuclear Information System (INIS)

    Schweickert, H.; Fehsenfeld, P.

    1995-01-01

    The reliability of industrial equip ment is substantially influenced by wear and corrosion; monitoring can prevent accidents and avoid down-time. One powerful tool is thin layer activation analysis (TLA) using accelerator systems. The information is used to improve mechanical design and material usage; the technology is used by many large companies, particularly in the automotive industry, e.g. Daimler Benz. A critical area of a machine component receives a thin layer of radioactivity by irradiation with charged particles from an accelerator - usually a cyclotron. The radioactivity can be made homogeneous by suitable selection of particle, beam energy and angle of incidence. Layer thickness can be varied from 20 microns to around 1 mm with different depth distributions; the position and size of the wear zone can be set to within 0.1 mm. The machine is then reassembled and operated so that wear can be measured. An example is a combustion engine comprising piston ring, cylinder wall, cooling water jacket and housing wall, where wear measurements on the cylinder wall are required in a critical zone around the dead-point of the piston ring. Proton beam bombardment creates a radioactive layer whose thickness is known accurately, and characteristic gamma radiation from this radioactive zone penetrates through the engine and is detected externally. Measurements can be made either of the activity removed from the surface, or of the (reduced) residual activity; wear measurement of the order of 10 -9 metres is possible

  8. Determination of Finasteride in Tablets by High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    K. Basavaiah

    2007-01-01

    Full Text Available A rapid, highly sensitive high performance liquid chromatographic method has been developed for the determination of finasteride(FNS in bulk drug and in tablets. FNS was eluted from a ODS C18 reversed phase column at laboratory temperature (30 ± 2°C with a mobile phase consisting of methanol and water (80+20 at a flow rate of 1 mL min-1 with UV detection at 225 nm. The retention time was ∼ 6.1 min and each analysis took not more than 10 min. Quantitation was achieved by measurement of peak area without using any internal standard. Calibration graph was linear from 2.0 to 30 μg mL-1 with limits of detection (LOD and quantification (LOQ being 0.2 and 0.6 μg mL-1, respectively. The method was validated according to the current ICH guidelines. Within-day co efficients of variation (CV ranged from 0.31 to 0.69% and between-day CV were in the range 1.2-3.2%. Recovery of FNS from the pharmaceutical dosage forms ranged from 97.89 – 102.9 with CV of 1.41-4.13%. The developed method was compared with the official method for FNS determination in its tablet forms.

  9. Experimental hydrophobicity parameters of perfluorinated alkylated substances from reversed-phase high performance liquid chromatography

    NARCIS (Netherlands)

    de Voogt, P.; Zurano, L.; Serné, P.; Haftka, J.J.H.

    2012-01-01

    Capacity factors of perfluorinated alkylated substances were obtained from isocratic reversed-phase high-performance liquid chromatography-mass spectrometry experiments at different organic modifier strengths of the mobile phase. The resulting capacity factor v. modifier strengths plots were

  10. Assay for dihydroorotase using high-performance liquid chromatography with radioactivity detection

    International Nuclear Information System (INIS)

    Mehdi, S.; Wiseman, J.S.

    1989-01-01

    An assay for measuring dihydroorotase activity was devised. Radiolabeled substrate and product were separated by high-performance liquid chromatography using a reverse-phase column with ion-pairing, and the radioactivity was quantitated by flow detection

  11. Development of a high-performance liquid chromatography method for the determination of florfenicol in animal feedstuffs.

    Science.gov (United States)

    Yang, JinJing; Sun, GuiZhi; Qian, MingRong; Huang, LingLi; Ke, XianBing; Yang, Bo

    2017-11-15

    An effective thin layer chromatography (TLC) purification procedure coupled to high-performance liquid chromatography (HPLC) method was developed for the determination of florfenicol (FF) in pig, chicken and fish feedstuffs. The feedstuff samples were extracted with ethyl acetate, defatted with n-hexane saturated with acetonitrile, and further purified by TLC. The chromatographic separation was performed on a Waters Symmetry C 18 column using an isocratic procedure with acetonitrile-water (35:65, v/v) at 0.6mL/min. The ultraviolet (UV) detector was set at a wavelength of 225nm. The FF concentrations in feedstuff samples were quantified using a standard curve. Good linear correlations (y=159075x-15054, r>0.9999) were achieved within the concentration range of 0.05-200μg/mL. The recoveries of FF spiked at levels of 1, 100 and 1000μg/g ranged from 80.6% to 105.3% with the intra-day and inter-day relative standard deviation (RSD) less than 9.3%. The limit of detection (LOD) and limit of quantitation (LOQ) were 0.02 and 0.06mg/kg for pig feedstuffs, 0.02 and 0.07mg/kg for chicken feedstuffs, and 0.02 and 0.05mg/kg for fish feedstuffs, respectively. This reliable, simple and cost-effective method could be applied to the routine monitoring of FF in animal feedstuffs. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Separation and quantitation of colour pigments of chili powder (Capsicum frutescens) by high-performance liquid chromatography-diode array detection.

    Science.gov (United States)

    Cserháti, T; Forgács, E; Morais, M H; Mota, T; Ramos, A

    2000-10-27

    The performance of reversed-phase thin-layer (RP-TLC) and reversed-phase high-performance liquid chromatography (RP-HPLC) was compared for the separation and determination of the colour pigments of chili (Capsicum frutescens) powder using a wide variety of eluent systems. No separation of pigments was achieved in RP-TLC, however, it was established that tetrahydrofuran shows an unusually high solvent strength. RP-HPLC using water-methanol-acetonitrile gradient elution separated the chili pigments in many fractions. Diode array detection (DAD) indicated that yellow pigments are eluted earlier than the red ones and chili powder contains more yellow pigments than common paprika powders. It was established that the very different absorption spectra of pigments make the use of DAD necessary.

  13. [Analysis of microalbuminuria with immunonephelometry and high performance liquid chromatography. Evaluation of new criteria].

    Science.gov (United States)

    Markó, Lajos; Molnár, Gergo Attila; Wagner, Zoltán; Koszegi, Tamás; Matus, Zoltán; Mohás, Márton; Kuzma, Mónika; Szijártó, István András; Wittmann, István

    2008-01-13

    Hypertension as well as type 2 diabetes mellitus is a major factor in population mortality. Both diseases damage the endothelium, the early sign of which is microalbuminuria, which can be screened by dipstick and can be diagnosed by using immuno-based and high performance liquid chromatography methods. Using high performance liquid chromatography, the non-immunoreactive albumin can be detected as well. The authors aimed at the examination of albuminuria in the case of immunonephelometrically negative patients with high performance liquid chromatography, in diabetic and hypertensive and non-diabetic hypertensive populations. The authors also wanted to compare the present (albumin-creatinine ratio: male: > or =2.5 mg/mmol, female: > or =3.5 mg/mmol) and a new criteria of the Heart Outcomes Prevention Evaluation study (patients without diabetes: immunological method, > or =0.7 mg/mmol; high performance liquid chromatography, > or =3.1 mg/mmol; individuals with diabetes: immunological method, > or =1.4 mg/mmol; high performance liquid chromatography, > or =5.2 mg/mmol) of microalbuminuria. Examination of fresh urines of 469 microalbuminuria negative patients by dipstick were performed by immunonephelometry. Patients, who were microalbuminuria negative by immunonephelometry as well, were further analyzed by high performance liquid chromatography using the Accumintrade mark Kit, based on size-exclusion chromatography. Three times higher albuminuria were found with high performance liquid chromatography than with immunonephelometry. The intraindividual coefficient of variation did not differ in the two methods (37 +/- 31% vs. 40 +/- 31%, p = 0.869; immunonephelometry vs. high performance liquid chromatography; mean +/- standard deviation). Using the present criteria for microalbuminuria, 43% of immunonephelometrically negative patients proved to be microalbuminuric by high performance liquid chromatography. Using the new criteria of the Heart Outcomes Prevention

  14. Quality evaluation of moluodan concentrated pill using high-performance liquid chromatography fingerprinting coupled with chemometrics.

    Science.gov (United States)

    Tao, Lingyan; Zhang, Qing; Wu, Yongjiang; Liu, Xuesong

    2016-12-01

    In this study, a fast and effective high-performance liquid chromatography method was developed to obtain a fingerprint chromatogram and quantitative analysis simultaneously of four indexes including gallic acid, chlorogenic acid, albiflorin and paeoniflorin of the traditional Chinese medicine Moluodan Concentrated Pill. The method was performed by using a Waters X-bridge C 18 reversed phase column on an Agilent 1200S high-performance liquid chromatography system coupled with diode array detection. The mobile phase of the high-performance liquid chromatography method was composed of 20 mmol/L phosphate solution and acetonitrile with a 1 mL/min eluent velocity, under a detection temperature of 30°C and a UV detection wavelength of 254 nm. After the methodology validation, 16 batches of Moluodan Concentrated Pill were analyzed by this high-performance liquid chromatography method and both qualitative and quantitative evaluation results were achieved by similarity analysis, principal component analysis and hierarchical cluster analysis. The results of these three chemometrics were in good agreement and all indicated that batch 10 and batch 16 showed significant differences with the other 14 batches. This suggested that the developed high-performance liquid chromatography method could be applied in the quality evaluation of Moluodan Concentrated Pill. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Thin-layer chromatography of ternary complexes of group-IIIA metals with 2-thenoyltrifluoroacetone and 2,2'-bipyridyl on cellulose layer

    Energy Technology Data Exchange (ETDEWEB)

    Chao, H E; Saitoh, K; Suzuki, N [Tohoku Univ., Sendai (Japan). Faculty of Science

    1980-11-11

    Normal phase thin-layer chromatographic behaviour of several ternary complexes of group-IIIA metals with 2-thenoyltrifluoroacetone (TTA) and 2,2'bipyridyl (bpy) has been investigated on cellulose layer. The ternary complexes of lanthanide metals show higher mutual separability than the complexes with TTA alone. Mutual separation of TTA complexes with La(III), Ce(III), Eu(III) or Y(III), Sc(III), Th(IV), and U(VI) has been successfully achieved by two-dimensional TLC, primarily with carbon tetrachloride-benzene (75:25) containing 0.02M TTA, and secondary with carbon tetrachloride-hexane (35:65) containing both 0.02M TTA and 0.02M bpy.

  16. Phytochemical Profile of Erythrina variegata by Using High-Performance Liquid Chromatography and Gas Chromatography-Mass Spectroscopy Analyses

    OpenAIRE

    Suriyavathana Muthukrishnan; Subha Palanisamy; Senthilkumar Subramanian; Sumathi Selvaraj; Kavitha Rani Mari; Ramalingam Kuppulingam

    2016-01-01

    Natural products derived from plant sources have been utilized to treat patients with numerous diseases. The phytochemical constituents present in ethanolic leaf extract of Erythrina variegata (ELEV) were identified by using high-performance liquid chromatography (HPLC) and gas chromatography-mass spectroscopy (GC-MS) analyses. Shade dried leaves were powdered and extracted with ethanol for analyses through HPLC to identify selected flavonoids and through GC-MS to identify other molecules. Th...

  17. Characterization of natural organic colorants in historical and art objects by high-performance liquid chromatography.

    Science.gov (United States)

    Pauk, Volodymyr; Barták, Petr; Lemr, Karel

    2014-12-01

    High-performance liquid chromatography plays an important role in analysis of historical organic colorants. A number of papers have been published in this field over the last 30 years. Classification of the most commonly used natural dyes and an overview of high-performance liquid chromatography methods with main focus on recent works (2008 to the beginning of 2014) are provided. The review deals with an entire analytical protocol covering sample preparation, chromatographic separation, and suitable detection (UV/visible and fluorescent spectroscopy and mass spectrometric techniques). High-performance liquid chromatography has been successfully used in the complete characterization of some organic dyestuffs present in historical and art objects. The possibilities and difficulties for identification of natural sources of historical colorants are also discussed. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. High-performance liquid chromatography of oligoguanylates at high pH

    Science.gov (United States)

    Stribling, R.; Deamer, D. (Principal Investigator)

    1991-01-01

    Because of the stable self-structures formed by oligomers of guanosine, standard high-performance liquid chromatography techniques for oligonucleotide fractionation are not applicable. Previously, oligoguanylate separations have been carried out at pH 12 using RPC-5 as the packing material. While RPC-5 provides excellent separations, there are several limitations, including the lack of a commercially available source. This report describes a new anion-exchange high-performance liquid chromatography method using HEMA-IEC BIO Q, which successfully separates different forms of the guanosine monomer as well as longer oligoguanylates. The reproducibility and stability at high pH suggests a versatile role for this material.

  19. 薄层色谱法检测海州常山桐树叶中肾上腺素的含量%Identification and quantification of adrenaline from the leaves of Clerodendrum phlomidis using thin-layer chromatography

    Institute of Scientific and Technical Information of China (English)

    Muthu Kumaradoss Mohan Maruga Raja; Ruchi Sureshbhai Patel; Shri Hari Mishra

    2011-01-01

    @@ In the Ayurvedic, Siddha and other medical systems of India, products derived from herbs,used either as active ingredients or as adjuvants,hold paramount importance as alternative medi-cines.However, their standardization poses a great challenge.Correct identification of these drugs is often problematic in both their complete form as well as in powder form, since these medicinal plants are known by a variety of vernacular names and frequently many medicinal plants share the same name.Comparative thin-layer chromatography (co-TLC) with chemical or biological marker compounds can be used to standardize raw materials.Moreover, due to its simplicity, accuracy, cost effectiveness and rapidity, TLC is often used as an alternative to other chromatographic tech-niques for quantifying plant products.

  20. Ad-hoc surface-enhanced Raman spectroscopy methodologies for the detection of artist dyestuffs: thin layer chromatography-surface enhanced Raman spectroscopy and in situ on the fiber analysis.

    Science.gov (United States)

    Brosseau, Christa L; Gambardella, Alessa; Casadio, Francesca; Grzywacz, Cecily M; Wouters, Jan; Van Duyne, Richard P

    2009-04-15

    Tailored ad-hoc methods must be developed for successful identification of minute amounts of natural dyes on works of art using Surface-Enhanced Raman Spectroscopy (SERS). This article details two of these successful approaches using silver film over nanosphere (AgFON) substrates and silica gel coupled with citrate-reduced Ag colloids. The latter substrate functions as the test system for the coupling of thin-layer chromatography and SERS (TLC-SERS), which has been used in the current research to separate and characterize a mixture of several artists' dyes. The poor limit of detection of TLC is overcome by coupling with SERS, and dyes which co-elute to nearly the same spot can be distinguished from each other. In addition, in situ extractionless non-hydrolysis SERS was used to analyze dyed reference fibers, as well as historical textile fibers. Colorants such as alizarin, purpurin, carminic acid, lac dye, crocin, and Cape jasmine were thus successfully identified.

  1. Determination of the recognition site for adenine-specific methylase of Shigella sonnei 47 by hydazinolysis of DNA, followed by separation of the purine oligonucleotides by thin-layer chromatography on DEAE-cellulose

    International Nuclear Information System (INIS)

    Lopatina, N.G.; Kirnos, M.D.; Suchkov, S.V.; Vanyushin, B.F.; Nikol'skaya, I.I.; Debov, S.S.

    1985-01-01

    A method has been developed for the separation of oligopurine units according to length and composition by two-dimensional thin-layer chromatography on plates with DEAE-cellulose, permitting a comparative analysis of the content of various purine isopliths in DNA of different origin. In the case of the analysis of methylated DNA, the method permits a comparison of the substrate specificity of various enzymes of methylation of the adenine residues in DNA. In conjunction with enzymatic treatment of labeled methylated isopliths, the method permits determination of the methylatable sequence and in a number of cases an ascertainment of the recognition site for adenine-specific methylase as a whole. The proposed method was used to establish the fact that the methylase Ssol recognizes the sequence 5'...G-A-A-T-T-C...3' and methylates the adenine residue closest to its 5'-end

  2. Determination of low molecular weight thiols using monobromobimane fluorescent labeling and high-performance liquid chromatography

    Science.gov (United States)

    Fahey, Robert C.; Newton, Gerald L.

    1988-01-01

    Methods are described for the preparation and high-performance liquid chromatography (HPLC) analysis of monobromobimane derivatives of low molecular weight thiols in extracts of biological samples. Typical problems encountered in the development and application of these methods are discussed. Analysis of mung bean extract is used as an example.

  3. Sugar Determination in Foods with a Radially Compressed High Performance Liquid Chromatography Column.

    Science.gov (United States)

    Ondrus, Martin G.; And Others

    1983-01-01

    Advocates use of Waters Associates Radial Compression Separation System for high performance liquid chromatography. Discusses instrumentation and reagents, outlining procedure for analyzing various foods and discussing typical student data. Points out potential problems due to impurities and pump seal life. Suggests use of ribose as internal…

  4. High-Performance Liquid Chromatography in the Undergraduate Chemical Engineering Laboratory

    Science.gov (United States)

    Frey, Douglas D.; Guo, Hui; Karnik, Nikhila

    2013-01-01

    This article describes the assembly of a simple, low-cost, high-performance liquid chromatography (HPLC) system and its use in the undergraduate chemical engineering laboratory course to perform simple experiments. By interpreting the results from these experiments students are able to gain significant experience in the general method of…

  5. Serum Protein Profile Study of Clinical Samples Using High Performance Liquid Chromatography-Laser Induced Fluorescence

    DEFF Research Database (Denmark)

    Karemore, Gopal Raghunath; Ukendt, Sujatha; Rai, Lavanya

    2009-01-01

    The serum protein profiles of normal subjects, patients diagnosed with cervical cancer, and oral cancer were recorded using High Performance Liquid Chromatography combined with Laser Induced Fluorescence detection (HPLC-LIF). Serum protein profiles of the above three classes were tested for estab...

  6. Nitrate and nitrite content in bottled beverages by ion-pair high-performance liquid chromatography.

    Science.gov (United States)

    Song, Yang; Deng, Gui-Fang; Xu, Xiang-Rong; Chen, Yong-Hong; Chen, Feng; Li, Hua-Bin

    2013-01-01

    Nitrate and nitrite levels in six types of beverages--total of 292 individual samples from 73 brands (four bottles each)--from Guangzhou city in China were evaluated by ion-pair high-performance liquid chromatography. All samples contained nitrate. Nitrate and nitrite ranges were 0.43-46.08 and safety of Chinese bottled beverages.

  7. DETERMINATION OF CHLOROPHEONIS, NITROPHENOIS AND METHYLPHENOIS IN GROUND-WATER SAMPLES USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

    Science.gov (United States)

    A high performance liquid chromatography (HPLC) method was developed to quantitatively determine phenolic compounds and their isomers in aqueous samples. The HPLC method can analyze a mixture of 15 contaminants in the same analytical run with an analysis time of 25 minutes. The...

  8. DETERMINATION OF CHLOROPHENOLS, NITROPHENOLS, AND METHYLPHENOLS IN GROUND-WATER SAMPLES USING HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

    Science.gov (United States)

    A high performance liquid chromatography (HPLC) method was developed to quantitatively determine phenolic compounds and their isomers in aqueous samples. The HPLC method can analyze a mixture of 15 contaminants in the same analytical run with an analysis time of 25 minutes. The...

  9. Coupling nanoliter high-performance liquid chromatography to inductively coupled plasma mass spectrometry for arsenic speciation.

    Science.gov (United States)

    Cheng, Heyong; Shen, Lihuan; Liu, Jinhua; Xu, Zigang; Wang, Yuanchao

    2018-04-01

    Nanoliter high-performance liquid chromatography shows low consumption of solvents and samples, offering one of the best choices for arsenic speciation in precious samples in combination with inuctively coupled plasma mass spectrometry. A systematic investigation on coupling nanoliter high-performance liquid chromatography to inductively coupled plasma mass spectrometry from instrument design to injected sample volume and mobile phase was performed in this study. Nanoflow mobile phase was delivered by flow splitting using a conventional high-pressure pump with reuse of mobile phase waste. Dead volume was minimized to 60 nL for the sheathless interface based on the previously developed nanonebulizer. Capillary columns for nanoliter high-performance liquid chromatography were found to be sensitive to sample loading volume. An apparent difference was also found between the mobile phases for nanoliter and conventional high-performance liquid chromatography. Baseline separation of arsenite, arsenate, monomethylarsenic, and dimethylarsenic was achieved within 11 min on a 15 cm C 18 capillary column and within 12 min on a 25 cm strong anion exchange column. Detection limits of 0.9-1.8 μg/L were obtained with precisions variable in the range of 1.6-4.2%. A good agreement between determined and certified values of a certified reference material of human urine (GBW 09115) validated its accuracy along with good recoveries (87-102%). © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. High-performance liquid chromatography determination of dapsone, monoacetyldapsone, and pyrimethamine in filter paper blood spots

    DEFF Research Database (Denmark)

    Rønn, A M; Lemnge, M M; Angelo, H R

    1995-01-01

    A high-performance liquid chromatography method for the simultaneous analysis of dapsone (DDS), the major metabolite of DDS, monoacetyldapsone (MADDS), and pyrimethamine (PYR) was modified for capillary blood samples obtained by finger prick and dried on filter paper. Limit of quantitation using...

  11. Simultaneous analysis of small organic acids and humic acids using high performance size exclusion chromatography

    NARCIS (Netherlands)

    Qin, X.P.; Liu, F.; Wang, G.C.; Weng, L.P.

    2012-01-01

    An accurate and fast method for simultaneous determination of small organic acids and much larger humic acids was developed using high performance size exclusion chromatography. Two small organic acids, i.e. salicylic acid and 2,3-dihydroxybenzoic acid, and one purified humic acid material were used

  12. Extraction and Purification of Glucoraphanin by Preparative High-Performance Liquid Chromatography (HPLC)

    Science.gov (United States)

    Lee, Iris; Boyce, Mary C.

    2011-01-01

    A student activity that focuses on the isolation of glucoraphanin from broccoli using preparative high-performance liquid chromatography (HPLC) is presented here. Glucoraphanin is a glucosinolate, whose byproducts are known to possess anticancer properties. It is present naturally at high levels in broccoli and other "Brassica" vegetables. This…

  13. An Advanced, Interactive, High-Performance Liquid Chromatography Simulator and Instructor Resources

    Science.gov (United States)

    Boswell, Paul G.; Stoll, Dwight R.; Carr, Peter W.; Nagel, Megan L.; Vitha, Mark F.; Mabbott, Gary A.

    2013-01-01

    High-performance liquid chromatography (HPLC) simulation software has long been recognized as an effective educational tool, yet many of the existing HPLC simulators are either too expensive, outdated, or lack many important features necessary to make them widely useful for educational purposes. Here, a free, open-source HPLC simulator is…

  14. An on-line high performance liquid chromatography-crocin bleaching assay for detection of antioxidants

    NARCIS (Netherlands)

    Bountagkidou, O.; Klift, van der E.J.C.; Tsimidou, M.Z.; Ordoudi, S.A.; Beek, van T.A.

    2012-01-01

    An on-line HPLC (high performance liquid chromatography) method for the rapid screening of individual antioxidants in mixtures was developed using crocin as a substrate (i.e. oxidation probe) and 2,2'-azobis(2-amidinopropane dihydrochloride (AAPH)) in phosphate buffer (pH 7.5) as a radical

  15. Physicochemical, thin layer and gas-liquid chromatographic ...

    African Journals Online (AJOL)

    DELL

    2012-05-22

    May 22, 2012 ... PEG (3 m × 3 mm I. D.) was used for gas chromatography. Physicochemical analysis ... subjected to thin layer chromatography on plates (20 × 20 cm) having 0.25 mm thick silica gel ..... Headspace solid- phase microextraction ...

  16. Chemical fingerprint of Ganmaoling granule by double-wavelength ultra high performance liquid chromatography and ultra high performance liquid chromatography with quadrupole time-of-flight mass spectrometry.

    Science.gov (United States)

    Lou, Qiong; Ye, Xiaolan; Zhou, Yingyi; Li, Hua; Song, Fenyun

    2015-06-01

    A method incorporating double-wavelength ultra high performance liquid chromatography with quadrupole time-of-flight mass spectrometry was developed for the investigation of the chemical fingerprint of Ganmaoling granule. The chromatographic separations were performed on an ACQUITY UPLC HSS C18 column (2.1 × 50 mm, 1.8 μm) at 30°C using gradient elution with water/formic acid (1%) and acetonitrile at a flow rate of 0.4 mL/min. A total of 11 chemical constituents of Ganmaoling granule were identified from their molecular weight, UV spectra, tandem mass spectrometry data, and retention behavior by comparing the results with those of the reference standards or literature. And 25 peaks were selected as the common peaks for fingerprint analysis to evaluate the similarities among 25 batches of Ganmaoling granule. The results of principal component analysis and orthogonal projection to latent structures discriminant analysis showed that the important chemical markers that could distinguish the different batches were revealed as 4,5-di-O-caffeoylquinic acid, 3,5-di-O-caffeoylquinic acid, and 4-O-caffeoylquinic acid. This is the first report of the ultra high performance liquid chromatography chemical fingerprint and component identification of Ganmaoling granule, which could lay a foundation for further studies of Ganmaoling granule. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Determination of chloroacetanilide herbicide metabolites in water using high-performance liquid chromatography-diode array detection and high-performance liquid chromatography/mass spectrometry

    Science.gov (United States)

    Hostetler, K.A.; Thurman, E.M.

    2000-01-01

    Analytical methods using high-performance liquid chromatography-diode array detection (HPLC-DAD) and high-performance liquid chromatography/mass spectrometry (HPLC/MS) were developed for the analysis of the following chloroacetanilide herbicide metabolites in water: alachlor ethanesulfonic acid (ESA); alachlor oxanilic acid; acetochlor ESA; acetochlor oxanilic acid; metolachlor ESA; and metolachlor oxanilic acid. Good precision and accuracy were demonstrated for both the HPLC-DAD and HPLC/MS methods in reagent water, surface water, and ground water. The average HPLC-DAD recoveries of the chloroacetanilide herbicide metabolites from water samples spiked at 0.25, 0.5 and 2.0 ??g/l ranged from 84 to 112%, with relative standard deviations of 18% or less. The average HPLC/MS recoveries of the metabolites from water samples spiked at 0.05, 0.2 and 2.0 ??g/l ranged from 81 to 118%, with relative standard deviations of 20% or less. The limit of quantitation (LOQ) for all metabolites using the HPLC-DAD method was 0.20 ??g/l, whereas the LOQ using the HPLC/MS method was at 0.05 ??g/l. These metabolite-determination methods are valuable for acquiring information about water quality and the fate and transport of the parent chloroacetanilide herbicides in water. Copyright (C) 2000 Elsevier Science B.V.

  18. Rapid screening method to study the reactivity of UV filter substances towards skin proteins by high-performance thin-layer chromatography.

    Science.gov (United States)

    Stiefel, C; Schwack, W

    2013-12-01

    Most UV filters used in sunscreens and other cosmetic products contain carbonyl groups, which generally are able to react with peptides or free amino acids of the human skin. To estimate their reactivity, we studied different prominent UV filter substances, octocrylene, ethylhexyl salicylate, 4-t-butyl-4'-methoxydibenzoylmethane, ethylhexyl methoxycinnamate, benzophenone-3, hydroxymethylbenzoyl sulphonic acid, octyldimethyl p-aminobenzoic acid, 3-benzylidene camphor, 4-methylbenzylidene camphor, diethylhexyl butamido triazone and ethylhexyl triazone. A simple screening method using an amino HPTLC plate as protein model was established. The influence of different reaction conditions like heating and irradiation was determined. The ketones BP-3, HMBS and BM-DBM revealed the highest binding rates after both irradiation and heating. After 1 h of irradiation, 82%, 28% and 96%, respectively, were bonded to the amino phase, while heating resulted in values of 52%, 36% and 16%. For BP-3 and HMBS, even storage in the dark at room temperature resulted in a low binding. Contrarily, for the two camphor derivatives 3-BC and 4-MBC, only irradiation led to a slightly turnover. UV filters with ester groups also showed a different behaviour depending on their main skeleton. While OCR especially reacted under heating with the amino phase, resulting in 36% of bound species after one hour, UV irradiation particularly encouraged a reaction of the other esters. After 1 h irradiation, 15% of EHMC, 38% of EHS and 48% of OD-PABA were bonded to the amino groups of the HPTLC plate, whereas the reactivity of the two triazones, EHT and DEBT, was comparatively low. Especially the UV filters BP-3, BM-DBM, HMBS, EHMC or OCR, which are commonly known to cause contact dermatitis, showed a high tendency to form adducts with the amino layer. Thus, the amino plate seems to be a proper tool to screen for skin sensitizers. © 2013 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

  19. Validation of interventional fiber optic spectroscopy with MR Spectroscopy, MAS-NMR spectroscopy, high-performance thin-layer chromatography, and histopathology for accurate hepatic fat quantification

    NARCIS (Netherlands)

    Nachabé, R.; Hoorn, J.W.A. van der; Molengraaf, R. van de; Lamerichs, R.; Pikkemaat, J.; Sio, C.F.; Hendriks, B.H.W.; Sterenborg, H.J.C.M.

    2012-01-01

    Objectives: To validate near-infrared (NIR)-based optical spectroscopy measurements of hepatic fat content using a minimally invasive needle-like probe with integrated optical fibers, enabling real-time feedback during percutaneous interventions. The results were compared with magnetic resonance

  20. Simultaneous Detection of Three Phosphodiesterase Type 5 Inhibitors and Eight of Their Analogs in Lifestyle Products and Screening for Adulterants by High-Performance Thin-Layer Chromatography.

    Science.gov (United States)

    Do, Tiên T K; Theocharis, Grigorios; Reich, Eike

    2015-01-01

    An HPTLC method is proposed to permit effective screening for the presence of three phosphodiesterase type 5 inhibitors (PDE5-Is; sildenafil, vardenafil, and tadalafil) and eight of their analogs (hydroxyacetildenafil, homosildenafil, thiohomosildenafil, acetildenafil, acetaminotadalafil, propoxyphenyl hydroxyhomosildenafil, hydroxyhomosildenafil, and hydroxythiohomosildenafil) in finished products, including tablets, capsules, chocolate, instant coffee, syrup, and chewing gum. For all the finished products, the same simple sample preparation may be applied: ultrasound-assisted extraction in 10 mL methanol for 30 min followed by centrifugation. The Rf values of individual HPTLC bands afford preliminary identification of potential PDE5-Is. Scanning densitometry capabilities enable comparison of the unknown UV spectra with those of known standard compounds and allow further structural insight. Mass spectrometric analysis of the material derived from individual zones supplies an additional degree of confidence. Significantly, the proposed screening technique allows focus on the already known PDE5 Is and provides a platform for isolation and chemical categorization of the newly-synthesized analogs. Furthermore, the scope could be expanded to other therapeutic categories (e.g., analgesics, antidiabetics, and anorexiants) that are occasionally coadulterated along with the PDE5-Is. The method was successfully applied to screening of 45 commercial lifestyle products. Of those, 31 products tested positive for at least one illegal component (sildenafil, tadalafil, propoxyphenyl hydroxyhomosildenafil, or dimethylsildenafil).

  1. Stability studies of therapeutic 131I-meta-iodobenzylguanidine (131I-mIBG) using high performance liquid chromatography

    International Nuclear Information System (INIS)

    Murhekar, V.V.; Mathur, Anupam; Pilkhwal, Neelam S.; Prabhakar, G.; Padmanabhan, D.; Sachdev, S.S.

    2014-01-01

    BRIT is a manufacturer and supplier of therapeutic doses (100 mCi) of the radiopharmaceutical 131 I-mIBG to various nuclear medicine centers in India. The therapeutic formulation is of high radioactive concentration (>10 mCi/ml) and is thus prone to radiolytic damage during transport, storage until administration. Earlier stability studies at this laboratory were done using conventional methods like Thin Layer Chromatography (TLC) which has an inherent limitation in terms of resolution. In view of this, a suitable HPLC method has been developed and the stability of therapeutic 131 I-mIBG was monitored at various conditions

  2. Simultaneous determination of atorvastatin calcium and olmesartan medoxomil in a pharmaceutical formulation by reversed phase high-performance liquid chromatography, high-performance thin-layer chromatography, and UV spectrophotometric methods.

    Science.gov (United States)

    Soni, Hiral; Kothari, Charmy; Khatri, Deepak; Mehta, Priti

    2014-01-01

    Validated RP-HPLC, HPTLC, and UV spectrophotometric methods have been developed for the simultaneous determination of atorvastatin calcium (ATV) and olmesartan medoxomil (OLM) in a pharmaceutical formulation. The RP-HPLC separation was achieved on a Kromasil C18 column (250 x 4.6 mm, 5 microm particle size) using 0.01 M potassium dihydrogen o-phosphate (pH 4 adjusted with o-phosphoric acid)-acetonitrile (50 + 50, v/v) as the mobile phase at a flow rate of 1.5 mL/min. Quantification was achieved by UV detection at 276 nm. The HPTLC separation was achieved on precoated silica gel 60F254 plates using chloroform-methanol-acetonitrile (4 + 2+ 4, v/v/v) mobile phase. Quantification was achieved with UV detection at 276 nm. The UV-Vis spectrophotometric method was based on the simultaneous equation method that involves measurement of absorbance at two wavelengths, i.e., 255 nm (lambda max of OLM) and 246.2 nm (lambda max of ATV) in methanol. All three methods were validated as per International Conference on Harmonization guidelines. The proposed methods were simple, precise, accurate, and applicable for the simultaneous determination of ATV and OLM in a marketed formulation. The results obtained by applying the proposed methods were statistically analyzed and were found satisfactory.

  3. Identification of Ginkgo biloba supplements adulteration using high performance thin layer chromatography and ultra high performance liquid chromatography-diode array detector-quadrupole time of flight-mass spectometry

    Science.gov (United States)

    Ginkgo biloba is one of the most widely sold herbal supplements and medicines in the world. Its popularity stems to have a positive effect on memory and the circulatory system in clinical studies. As ginkgo popularity increased, non-proprietary extracts were introduced claiming to have similar phyto...

  4. Combined high-performance liquid chromatography/32P-postlabeling assay of N7-methyldeoxyguanosine

    International Nuclear Information System (INIS)

    Shields, P.G.; Povey, A.C.; Wilson, V.L.; Weston, A.; Harris, C.C.

    1990-01-01

    A highly sensitive and specific assay for the detection of N7-methyl-2'-deoxyguanosine (N7methyldG) has been developed by combining high-performance liquid chromatography, 32 P-postlabeling, and nucleotide chromatography. Separation of normal nucleotides and adducts by high-performance liquid chromatography and then combining a portion of 2'-deoxyguanosine to the N7methyldG allows for quantitation using an internal standard. The directly determined molar ratio is not subject to errors in digestion, variable ATP-specific activity, or assumptions in relative adduct-labeling efficiency. The detection limit was one N7methyldG adduct in 10(7) unmodified 2'-deoxyguanosine bases. N7methyldG adducts have been detected in 5 human lung samples in which O6-methyl-2'-deoxyguanosine adducts had been previously determined. The mean ratio of N7methyldG to O6-methyl-2'-deoxyguanosine was determined to be approximately 10. The current assay complements the high-performance liquid chromatography/ 32 P-postlabeling assay for O6-methyl-2'-deoxyguanosine and increases the detection sensitivity of DNA methylated by exogenous alkylating agents

  5. Characterization of Extracellular Vesicles by Size-Exclusion High-Performance Liquid Chromatography (HPLC).

    Science.gov (United States)

    Huang, Tao; He, Jiang

    2017-01-01

    Extracellular vesicles (EVs) have recently attracted substantial attention due to the potential diagnostic and therapeutic relevance. Although a variety of techniques have been used to isolate and analyze EVs, it is still far away from satisfaction. Size-exclusion chromatography (SEC), which separates subjects by size, has been widely applied in protein purification and analysis. The purpose of this chapter is to show the applications of size-exclusion high-performance liquid chromatography (HPLC) as methods for EV characterization of impurities or contaminants of small size, and thus for quality assay for the purity of the samples of EVs.

  6. Comparison of ultra-high performance supercritical fluid chromatography and ultra-high performance liquid chromatography for the separation of spirostanol saponins.

    Science.gov (United States)

    Zhu, Ling-Ling; Zhao, Yang; Xu, Yong-Wei; Sun, Qing-Long; Sun, Xin-Guang; Kang, Li-Ping; Yan, Ren-Yi; Zhang, Jie; Liu, Chao; Ma, Bai-Ping

    2016-02-20

    Spirostanol saponins are important active components of some herb medicines, and their isolation and purification are crucial for the research and development of traditional Chinese medicines. We aimed to compare the separation of spirostanol saponins by ultra-high performance supercritical fluid chromatography (UHPSFC) and ultra-high performance liquid chromatography (UHPLC). Four groups of spirostanol saponins were separated respectively by UHPSFC and UHPLC. After optimization, UHPSFC was performed with a HSS C18 SB column or a Diol column and with methanol as the co-solvent. A BEH C18 column and mobile phase containing water (with 0.1% formic acid) and acetonitrile were used in UHPLC. We found that UHPSFC could be performed automatically and quickly. It is effective in separating the spirostanol saponins which share the same aglycone and vary in sugar chains, and is very sensitive to the number and the position of hydroxyl groups in aglycones. However, the resolution of spirostanol saponins with different aglycones and the same sugar moiety by UHPSFC was not ideal and could be resolved by UHPLC instead. UHPLC is good at differentiating the variation in aglycones, and is influenced by double bonds in aglycones. Therefore, UHPLC and UHPSFC are complementary in separating spirostanol saponins. Considering the naturally produced spirostanol saponins in herb medicines are different both in aglycones and in sugar chains, a better separation can be achieved by combination of UHPLC and UHPSFC. UHPSFC is a powerful technique for improving the resolution when UHPLC cannot resolve a mixture of spirostanol saponins and vice versa. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Identification and Quantitation of Asparagine and Citrulline Using High-Performance Liquid Chromatography (HPLC)

    OpenAIRE

    Bai, Cheng; Reilly, Charles C.; Wood, Bruce W.

    2007-01-01

    High-performance liquid chromatography (HPLC) analysis was used for identification of two problematic ureides, asparagine and citrulline. We report here a technique that takes advantage of the predictable delay in retention time of the co-asparagine/citrulline peak to enable both qualitative and quantitative analysis of asparagine and citrulline using the Platinum EPS reverse-phase C18 column (Alltech Associates). Asparagine alone is eluted earlier than citrulline alone, but when both of them...

  8. Simultaneous Determination of Four Preservatives in Foodstuffs by High Performance Liquid Chromatography

    OpenAIRE

    Mohammad Faraji; Farzaneh Rahbarzare

    2016-01-01

    Background and objectives:  High concentration of preservatives in food may result in gastrointestinal disturbances whereby some patients suffering from asthma, rhinitis, or urticaria. The aim of this study is the introduction and optimization a new method for simultaneous determination of four preservatives (SB, PS, MP, PP) in foodstuff by high performance liquid chromatography. Materials and methods: Important factors in extraction, separation and determination process were optimiz...

  9. Core-Shell Columns in High-Performance Liquid Chromatography: Food Analysis Applications

    OpenAIRE

    Preti, Raffaella

    2016-01-01

    The increased separation efficiency provided by the new technology of column packed with core-shell particles in high-performance liquid chromatography (HPLC) has resulted in their widespread diffusion in several analytical fields: from pharmaceutical, biological, environmental, and toxicological. The present paper presents their most recent applications in food analysis. Their use has proved to be particularly advantageous for the determination of compounds at trace levels or when a large am...

  10. Enantioselective analysis of drugs: contributions of high-performance liquid chromatography and capillary electrophoresis

    OpenAIRE

    Bonato, Pierina Sueli; Jabor, Valquíria Aparecida Polisel; Gaitani, Cristiane Masetto de

    2005-01-01

    The demand for analytical methods suitable for accurate and reproducible determination of drug enantiomers has increased significantly in the last years. High-performance liquid chromatography (HPLC) using chiral stationary phases and capillary electrophoresis (CE) are the most important techniques used for this purpose. In this paper, the fundamental aspects of chiral separations using both techniques are presented. Some important aspects for the development of enantioselective methods, part...

  11. RECENT ADVANCES IN ULTRA-HIGH PERFORMANCE LIQUID CHROMATOGRAPHY FOR THE ANALYSIS OF TRADITIONAL CHINESE MEDICINE

    Science.gov (United States)

    Huang, Huilian; Liu, Min; Chen, Pei

    2014-01-01

    Traditional Chinese medicine has been widely used for the prevention and treatment of various diseases for thousands of years in China. Ultra-high performance liquid chromatography (UHPLC) is a relatively new technique offering new possibilities. This paper reviews recent developments in UHPLC in the separation and identification, fingerprinting, quantification, and metabolism of traditional Chinese medicine. Recently, the combination of UHPLC with MS has improved the efficiency of the analysis of these materials. PMID:25045170

  12. Toxic Compounds Analysis With High Performance Liquid Chromatography Detected By Electro Chemical Detector (Ecd)

    OpenAIRE

    Hideharu Shintaniq

    2014-01-01

    The principal area of application of high performance liquid chromatography-electrochemical detector (HPLC-ECD) has been in the analysis of naturally-occurring analytes, such as catecholamines, and pharmaceuticals in biological samples, HPLC-ECD has also applied to the analysis of pesticides and other analytes of interest to the toxicologist. In this paper, toxic area is described. In these, ammatoxins, aromatic amine, nitro-compounds, algal toxins, fungal toxins, pesticides, veterinary drug ...

  13. Resolution of the stereoisomers of baclofen by high performance liquid chromatography

    International Nuclear Information System (INIS)

    Weatherby, R.P.; Allan, R.D.; Johnston, G.A.R.

    1984-01-01

    The GABA analogue baclofen [3-(p-chlorophenyl)-4-aminobutanoic acid] has stereospecific actions on the peripheral and central nervous systems. This paper describes the resolution of tritium-labelled baclofen by high performance liquid chromatography on a reverse-phase C18 column using a chiral mobile phase. The method, which may have general application to certain other GABA analogues, affords optically pure (+)- and (-)-baclofen labelled with tritium to high specific activity suitable for ligand binding and other neurochemical studies. (Auth.)

  14. Efficient methods for isolating five phytochemicals from Gentiana macrophylla using high-performance countercurrent chromatography.

    Science.gov (United States)

    Rho, Taewoong; Jung, Mila; Lee, Min Won; Chin, Young-Won; Yoon, Kee Dong

    2016-12-01

    Efficient high-performance countercurrent chromatography methods were developed to isolate five typical compounds from the extracts of Gentiana macrophylla. n-Butanol-soluble extract of G. macrophylla contained three hydrophilic iridoids, loganic acid (1), swertiamarin (2) and gentiopicroside (3), and a chromene derivative, macrophylloside D (4) which were successfully isolated by flow rate gradient (1.5 mL/min in 0-60 min, 5.0 mL/min in 60-120 min), and consecutive flow rate gradient HPCCC using n-butanol/0.1% aqueous trifluoroacetic acid (1:1, v/v, normal phase mode) system. The yields of 1-4 were 22, 16, 122, and 6 mg, respectively, with purities over 97% in a flow rate gradient high-performance countercurrent chromatography, and consecutive flow rate gradient high-performance countercurrent chromatography gave 1, 2, 3 (54, 41, 348 mg, respectively, purities over 97%) and 4 (13 mg, purity at 95%) from 750 mg of sample. The main compound in methylene chloride soluble extract, 2-methoxyanofinic acid, was successfully separated by n-hexane/ethyl acetate/methanol/water (4:6:4:6, v/v/v/v, flow-rate: 4 mL/min, reversed phase mode) condition. The structures of five isolates were elucidated by 1 H, 13 C NMR and ESI-Q-TOF-MS spectroscopic data which were compared with previously reported values. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Separation of enantiomers of new psychoactive substances by high-performance liquid chromatography.

    Science.gov (United States)

    Kadkhodaei, Kian; Forcher, Lisa; Schmid, Martin G

    2018-03-01

    New psychoactive substances are defined as compounds with consciousness-changing effects and have been developed simultaneously with classical drugs. They arise through structural modifications of illegal substances and are mainly produced to circumvent laws. Availability is simple, since new psychoactive substances can be purchased from the Internet. Among them many chemical drug compound classes are chiral and thus the two resulting enantiomers can differ in their effects. The aim of this study is to develop a suitable chiral high-performance liquid chromatography separation method for a broad spectrum of new psychoactive substances using cellulose tris(3,5-dichlorophenylcarbamate) as a chiral selector. Experiments were performed by high-performance liquid chromatography in normal-phase mode under isocratic conditions using ultraviolet detection. Direct separation was carried out on a high-performance liquid chromatography column (Lux® i-Cellulose-5, 3.5 μm, Phenomenex®), available since 2016. Excellent separation results were obtained for cathinones. After further optimization, even 47 instead of 39 out of 52 cathinones showed baseline separation. For amphetamine derivatives, satisfactory results were not achieved. Further, new psychoactive substances from other compound classes such as benzofuranes, thiophenes, phenidines, phenidates, morpholines, and ketamines were partially resolved, depending on the polarity and degree of substitution. All analytes, which were mainly purchased from the Internet, were proven to be traded as racemates. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Separation of hemagglutination-inhibiting immunoglobulin M antibody to rubella virus in human serum by high-performance liquid chromatography.

    OpenAIRE

    Kobayashi, N; Suzuki, M; Nakagawa, T; Matumoto, M

    1986-01-01

    High-performance liquid chromatography was successfully used to separate hemagglutination-inhibiting immunoglobulin M (IgM) rubella virus antibody from IgG rubella virus antibody in human serum. The fractionation by high-performance liquid chromatography was as effective as sucrose density gradient centrifugation in separating IgM antibody from IgG antibody.

  17. Cromatografia em camada delgada para o diagnóstico da intoxicação por aldicarb ("chumbinho" em cães e gatos Thin-layer chromatography for aldicarb poisoning diagnosis in dogs and cats

    Directory of Open Access Journals (Sweden)

    F.G. Xavier

    2007-10-01

    Full Text Available Avaliou-se a cromatografia em camada delgada (CCD como método de diagnóstico toxicológico para os casos de intoxicação por aldicarb em cães e gatos, utilizando-se 50 amostras de conteúdo gástrico obtidas durante a necropsia e 50 amostras de alimentos utilizados como iscas para intoxicar criminalmente os animais. Todas as amostras resultaram positivas para o aldicarb, mostrando ser a CCD uma técnica qualitativa eficiente, rápida e de baixo custo, com uso potencial na toxicologia veterinária forense.The present study concerns about the identification of aldicarb residues using thin-layer chromatography (TLC in 50 samples of gastric content obtained from the necropsy of dogs and cats and 50 samples of foods suspected of being used as baits. All samples resulted positive for aldicarb showing that the TLC is an efficient, fast and not expensive qualitative method for the detection of aldicarb, being useful for this purpose in the forensic veterinary toxicology.

  18. Development and evaluation of thin-layer chromatography-digital image-based analysis for the quantitation of the botanical pesticide azadirachtin in agricultural matrixes and commercial formulations: comparison with ELISA.

    Science.gov (United States)

    Tanuja, Penmatsa; Venugopal, Namburi; Sashidhar, Rao Beedu

    2007-01-01

    A simple thin-layer chromatography-digital image-based analytical method has been developed for the quantitation of the botanical pesticide, azadirachtin. The method was validated by analyzing azadirachtin in the spiked food matrixes and processed commercial pesticide formulations, using acidified vanillin reagent as a postchromatographic derivatizing agent. The separated azadirachtin was clearly identified as a green spot. The Rf value was found to be 0.55, which was similar to that of a reference standard. A standard calibration plot was established using a reference standard, based on the linear regression analysis [r2 = 0.996; y = 371.43 + (634.82)x]. The sensitivity of the method was found to be 0.875 microg azadirachtin. Spiking studies conducted at the 1 ppm (microg/g) level in various agricultural matrixes, such as brinjal, tomato, coffee, and cotton seeds, revealed the recoveries of azadirachtin in the range of 67-92%. Azadirachtin content of commercial neem formulations analyzed by the method was in the range of 190-1825 ppm (microg/mL). Further, the present method was compared with an immunoanalytical method enzyme-linked immonosorbent assay developed earlier in our laboratory. Statistical comparison of the 2 methods, using Fischer's F-test, indicated no significant difference in variance, suggesting that both methods are comparable.

  19. Assessing the detectability of antioxidants in two-dimensional high-performance liquid chromatography.

    Science.gov (United States)

    Bassanese, Danielle N; Conlan, Xavier A; Barnett, Neil W; Stevenson, Paul G

    2015-05-01

    This paper explores the analytical figures of merit of two-dimensional high-performance liquid chromatography for the separation of antioxidant standards. The cumulative two-dimensional high-performance liquid chromatography peak area was calculated for 11 antioxidants by two different methods--the areas reported by the control software and by fitting the data with a Gaussian model; these methods were evaluated for precision and sensitivity. Both methods demonstrated excellent precision in regards to retention time in the second dimension (%RSD below 1.16%) and cumulative second dimension peak area (%RSD below 3.73% from the instrument software and 5.87% for the Gaussian method). Combining areas reported by the high-performance liquid chromatographic control software displayed superior limits of detection, in the order of 1 × 10(-6) M, almost an order of magnitude lower than the Gaussian method for some analytes. The introduction of the countergradient eliminated the strong solvent mismatch between dimensions, leading to a much improved peak shape and better detection limits for quantification. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. Determination of ferulic acid and related compounds by thin layer ...

    African Journals Online (AJOL)

    The analysis of certain phenolic compounds from plants, and their chemical transformation with microorganisms or isolated enzymes, has application in the food and pharmaceutical industry. The rapid quantitative estimation of ferulic acid by thin layer chromatography is described by measurement of the area of the ...

  1. Evaluation of a thin-layer chromatographic technique for ...

    African Journals Online (AJOL)

    Methanol extracts of both fistula and bush samples were prepared and analysed by thin-layer chromatography. Chromatoplates, when visualised under ultraviolet light, revealed a number of fluorescent compounds, some of which were common in both the fistula and bush sample extracts. By comparing the presence of ...

  2. Comparison of two detection methods in thin layer chromatographic ...

    African Journals Online (AJOL)

    o-tolidine plus potassium iodide and photosynthesis inhibition detection methods were investigated for the analysis of three triazine herbicides (atrazine, ametryne, simazine) and two urea herbicides (diuron, metobromuron) in a coastal savanna soil using thin layer chromatography to compare the suitability of the two ...

  3. Physicochemical properties of silica gel coated with a thin layer of polyaniline (PANI) and its application in non-suppressed ion chromatography.

    Science.gov (United States)

    Sowa, Ireneusz; Kocjan, Ryszard; Wójciak-Kosior, Magdalena; Swieboda, Ryszard; Zajdel, Dominika; Hajnos, Mieczysław

    2013-10-15

    Physicochemical properties of a new sorbent and its potential application in non-suppressed ion chromatography (IC) have been investigated. The sorbent was obtained in a process of covering silica gel particles with a film of polyaniline (PANI). The properties of silica modified with polyaniline such as particle size, porosity, average quantity of polyaniline covering carrier and density of sorbent were determined. In our study the following methods were used: microscopic analysis, laser diffraction technique, combustion analysis, mercury porosimetry and helium pycnometry. Column with the newly obtained packing was used for the separation of inorganic anions. Optimized chromatographic system was successfully employed for analysis of iodide and bromide in selected pharmaceutical products (Bochnia salt and Iwonicz salt) applied in chronic respiratory disease. Analysis was carried out using 0.1M solution of HCl in mixture of methanol/water (50:50v/v) as a mobile phase; the flow rate was 0.3 mL min(-1), temperature was 24°C and λ=210 nm. Validation parameters such as correlation coefficient, RSD values, recovery, detection and quantification limits were found to be satisfactory. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Simultaneous Determination of Eight Bioactive Compounds in Dianthus superbus by High-performance Liquid Chromatography.

    Science.gov (United States)

    Yun, Bo-Ra; Yang, Hye Jin; Weon, Jin Bae; Lee, Jiwoo; Eom, Min Rye; Ma, Choong Je

    2016-05-01

    Dianthus superbus, one of traditional herbal medicine, is widely used to treat urethritis, carbuncles and carcinoma. A simultaneous determination method was established for controlling the quality of D. superbus using the eight compounds, (E)-methyl-4-hydroxy-4-(8a-methyl-3-oxodecahydronaphthalen-4a-yl) (1), diosmetin-7-O(2'',6''-di-O-α-L-rhamnopyranosyl)-β-D-glucopyranoside (2), vanillic acid (3), 4-hydroxyphenyl acetic acid (4), 4-methoxyphenyl acetic acid (5), (E)-4-methoxycinnamic acid (6), 3-methoxy-4-hydroxyphenylethanol (7), and methyl hydroferulate (8) isolated from D. superbus. This analysis method was developed using high performance liquid chromatography coupled with diode array detector with a Shishedo C18 column at a column temperature of 3°C. The mobile phase was composed of 0.1% trifluoroacetic acid in water and acetonitrile. The flow rate was 1 ml/min and detection wavelength was set at 205 nm and 280 nm. Validation was performed in order to demonstrate selectivity, accuracy and precision of the method. The calibration curves showed good linearity (R (2) > 0.99). The limits of detection and limits of quantification were within the ranges 0.0159-0.6205 μg/ml and 0.3210-1.8802 μg/ml, respectively. Moreover, the relative standard deviations of intra- and inter-day precision were both Dianthus superbus was established by high performance liquid chromatography-diode array detectorDeveloped analysis method is validated with linearity, precious and accuracyThe newly established method was successfully evaluated contents of eight compounds in 12 D. superbus samples (D.1.D.12) from various regions and compared. Abbreviations used: HPLC: High performance liquid chromatography, LOD: Limits of detection, LOQ: Limits of quantification, RSD: Relative standard deviation.

  5. Determination of alpha-Tocopherol (vitamin E) in irradiated garlic by high performance liquid chromatography (HPLC)

    International Nuclear Information System (INIS)

    Rios, Magda Dias Goncalves; Penteado, Marilene de Vuono Camargo

    2003-01-01

    The effects of 60 Co ionizing radiations in doses of 0, 75, 100, 150, 200 and 250Gy on garlic, upon the α-tocopherol concentration were studied. The α-tocopherol contents were established by high performance liquid chromatography (HPLC), after direct hexane extraction from the garlic samples. The α-tocopherol was determined through normal phase column, and mobile phase was composed by hexane: iso-propyl alcohol (99:01 v/v), with 2mL/min flow rate and fluorescence detector. It is statistically shown that an irradiation dose of up to 150 Gy does not affect the garlic α-tocopherol content. (author)

  6. 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....... The developed method is presently used for measurement of 1-hydroxypyrene in urine samples from workers exposed to a low airborne level of polycyclic aromatic hydrocarbons, generally less than 25 micrograms/m3. The urine samples of exposed workers (n = 122) showed a range of 1-hydroxypyrene from the limit...

  7. Application of Ionic Liquids in High Performance Reversed-Phase Chromatography

    Directory of Open Access Journals (Sweden)

    Wentao Bi

    2009-06-01

    Full Text Available Ionic liquids, considered “green” chemicals, are widely used in many areas of analytical chemistry due to their unique properties. Recently, ionic liquids have been used as a kind of novel additive in separation and combined with silica to synthesize new stationary phase as separation media. This review will focus on the properties and mechanisms of ionic liquids and their potential applications as mobile phase modifier and surface-bonded stationary phase in reversed-phase high performance liquid chromatography (RP-HPLC. Ionic liquids demonstrate advantages and potential in chromatographic field.

  8. Denaturing high-performance liquid chromatography mutation analysis in patients with reduced Protein S levels

    DEFF Research Database (Denmark)

    Bathum, Lise; Münster, Anna-Marie; Nybo, Mads

    2008-01-01

    diagnosis and risk estimation. The aim was to design a high-throughput genetic analysis based on denaturing high-performance liquid chromatography to identify sequence variations in the gene coding for Protein S. PATIENTS: In total, 55 patients referred to the Section of Thrombosis and Haemostasis, Odense......BACKGROUND: Patients with congenital Protein S deficiency have increased risk of venous thromboembolism. However, Protein S levels show large intra-individual variation and the biochemical assays have low accuracy and a high interlaboratory variability. Genetic analysis might aid in a more precise......, giving a precise diagnosis and subsequently a better risk estimation....

  9. High Performance Affinity Chromatography of Antithrombin III Based on Monodisperse Poly (glycidyl methacrylate) Beads

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    A new approach for the separation of antithrombin III with high performance affinity chromatography (HPAC) was described. A novel monodisperse,non-porous,cross-linked poly (glycidyl methacrylate) beads (PGMA) were used as the affinity support. With the water-soluble carbodiimide,heparin was linked covalently to amino-PGMA-beads,which was prepared by amination of PGMA. The adsorbent obtained exhibits high binding activity to antithrombin III (ATIII),good resolution and excellent mechanical properties and can be used under high flow rate.

  10. Hb A1c Separation by High Performance Liquid Chromatography in Hemoglobinopathies

    OpenAIRE

    Chandrashekar, Vani

    2016-01-01

    Hb A1c measurement is subject to interference by hemoglobin traits and this is dependent on the method used for determination. In this paper we studied the difference between Hb A1c measured by HPLC in hemoglobin traits and normal chromatograms. We also studied the correlation of Hb A1c with age. Hemoglobin analysis was carried out by high performance liquid chromatography. Spearman's rank correlation was used to study correlation between A1c levels and age. Mann-Whitney U test was used to st...

  11. High-performance ion-exchange chromatography of alkali metals with conductivity detection

    International Nuclear Information System (INIS)

    Ahmad, M.; Khan, A.R.

    1981-01-01

    High-performance ion-exchange chromatography of alkali metal and ammonium ions was studied using a conductivity meter as detector. Elution with 0.003 N mitric acid gave excellent resolution. Sensitivity levels, for a 200 micro litre injection, vary from 5 ppm for potassium to 0.1 ppm for lithium. A method to decrease retention times by reducing the exchange capacity of the cation exchange column used by loading it with calciumions, without affecting the resolation, has been described. Application of the method to water, soil and uranium dioxide samples has been demonstrated. (author)

  12. High-performance liquid chromatography of rat and mouse islet polypeptides

    DEFF Research Database (Denmark)

    Linde, S; Hansen, B; Welinder, B S

    1990-01-01

    After preparative high-performance liquid chromatography of mouse islet culture medium, concentrated on disposable C18 cartridges (Sep-Pak), an unexpected insulin immunoreactive peak eluting earlier than mouse insulin I and II was detected. Molecular mass determination by mass spectrometry...... on the buffer, the organic modifier and the procedure. In particular the use of methanol-trifluoroacetic acid resulted in extensive oxidation. The oxidation could be minimized by adding 2 mM dithiothreitol to the buffer and by degassing and/or nitrogen-bubbling of the buffer. Minimal formation of Met...

  13. Quantification of glibenclamide in cleaning samples of pharmaceutical equipment through high performance liquid chromatography

    International Nuclear Information System (INIS)

    Baeza Fonte, Alen Nils; Diaz Aguila, Elsa Eneida; Martinez Alfonso; Nancy

    2012-01-01

    to submit a selective analytical method for quantization of glibenclamide in cleaning samples of pharmaceutical equipment using high performance liquid chromatography. The mobile phase consisted of an equal mixing of acetonitrile/phosphate buffer KH 2 PO 4 ; with 0.037 mol/L concentration pH 5.25 and flow of 1.5 mL/min, in a Nucleosil 100 C8 column. Glibenclamide was injected with progesterone as internal standard and using an UV detector= 230 nm

  14. Determination of thebaine in water samples, biological fluids, poppy capsule, and narcotic drugs, using electromembrane extraction followed by high-performance liquid chromatography analysis.

    Science.gov (United States)

    Seidi, Shahram; Yamini, Yadollah; Heydari, Akbar; Moradi, Morteza; Esrafili, Ali; Rezazadeh, Maryam

    2011-09-09

    Opium determination is of great importance from toxicological and pharmaceutical standpoints. In present work, electromembrane extraction (EME) coupled with high-performance liquid chromatography (HPLC) and ultraviolet (UV) detection was developed for determination of thebaine as a natural alkaloid, in different matrices containing water, urine, poppy capsule, street heroine, and codeine tablet. Thebaine migrated from 3 mL of sample solutions, through a thin layer of 2-nitrophenyl octyl ether (NPOE) immobilized in the pores of a porous hollow fiber, and into a 15 μL acidic aqueous acceptor solution present inside the lumen of the fiber. The variables of interest, such as chemical composition of the organic liquid membrane, stirring speed, extraction time and voltage, pH of donor and acceptor phases and salt effect in the EME process were optimized. Under optimal conditions, thebaine was effectively extracted from different matrices with recoveries in the range of 45-55%, which corresponded to preconcentration factors in the range of 90-110. Good linearity was achieved for calibration curves with a coefficient of estimation higher than 0.997. Detection limits and intra-day precision (n=3) were less than 15 μg L(-1) and 8.9%, respectively. Copyright © 2011 Elsevier B.V. All rights reserved.

  15. Use of thin layer chromatography for the determination of radiochemical purity of radiopharmaceuticals in nuclear medicine services of Paraiba and Rio Grande do Norte, Brazil; Utilizacao da cromatografia em camada delgada para determinacao da pureza radioquimica de radiofarmacos em servicos de medicina nuclear da Paraiba e Rio Grande do Norte, Brasil

    Energy Technology Data Exchange (ETDEWEB)

    Andrade, W.G.; Santos, P.A.L.; Lima, F.R.A. [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Programa de Pos-Graduacao em Tecnologia Energetica; Lima, F.F., E-mail: wellington.gandrade@gmail.com [Centro Regional de Ciencias Nucleares do Nordeste (CRCN-NE/CNEN-PE), Recife, PE (Brazil)

    2012-07-01

    The paper chromatography and the thin layer chromatography are separation techniques in which the radioactive components migrate because of their affinity with the eluent (mobile phase) or stationary phase, respectively. In radiopharmaceuticals labeled with {sup 99m}Tc, besides its own radiopharmaceutical, {sup 99m}TcO{sup 4-} free and TcO{sub 2} can be identified and quantified. The evaluation of radiochemical purity of radiopharmaceuticals is essential to produce images free of artifacts as well as avoid unnecessary absorbed dose to the patient. Once they are managed in humans it is important and necessary that they undergo to strict quality control. Because of this, ANVISA in its 'Resolucao da Diretoria Colegiada (RDC) 38 of June 4th, 2008 states the obligation of performing a minimum of tests in nuclear medicine services routine prior to human administration. This work evaluated, by the method of thin layer chromatography (TLC), radiochemical purity, determined the pH of the radiopharmaceutical DEXTRAN- 500, DMSA, DTPA, PHYTATE, MDP, MIBI and Sn-Col used in nuclear medicine services in the states of Paraiba and Rio Grande do Norte - Brazil. The results show that the use of thin layer chromatography (TLC) as a standard method in routine of nuclear medicine services is possible, because it provides important data for the evaluation of radiochemical purity, allowing the exclusion of a radiopharmaceutical poorly marked. (author)

  16. Development and validation of ultra-high performance supercritical fluid chromatography method for determination of illegal dyes and comparison to ultra-high performance liquid chromatography method.

    Science.gov (United States)

    Khalikova, Maria A; Šatínský, Dalibor; Solich, Petr; Nováková, Lucie

    2015-05-18

    A novel simple, fast and efficient ultra-high performance supercritical fluid chromatography (UHPSFC) method was developed and validated for the separation and quantitative determination of eleven illegal dyes in chili-containing spices. The method involved a simple ultrasound-assisted liquid extraction of illegal compounds with tetrahydrofuran. The separation was performed using a supercritical fluid chromatography system and CSH Fluoro-Phenyl stationary phase at 70°C. The mobile phase was carbon dioxide and the mixture of methanol:acetonitrile (1:1, v/v) with 2.5% formic acid as an additive at the flow rate 2.0 mL min(-1). The UV-vis detection was accomplished at 500 nm for seven compounds and at 420 nm for Sudan Orange G, Butter Yellow, Fast Garnet GBC and Methyl Red due to their maximum of absorbance. All eleven compounds were separated in less than 5 min. The method was successfully validated and applied using three commercial samples of chili-containing spices - Chili sauce (Indonesia), Feferony sauce (Slovakia) and Mojo sauce (Spain). The linearity range of proposed method was 0.50-9.09 mg kg(-1) (r ≥ 0.995). The detection limits were determined as signal to noise ratio of 3 and were ranged from 0.15 mg kg(-1) to 0.60 mg kg(-1) (1.80 mg kg(-1) for Fast Garnet) for standard solution and from 0.25 mg kg(-1) to 1.00 mg kg(-1) (2.50 mg kg(-1) for Fast Garnet, 1.50 mg kg(-1) for Sudan Red 7B) for chili-containing samples. The recovery values were in the range of 73.5-107.2% and relative standard deviation ranging from 0.1% to 8.2% for within-day precision and from 0.5% to 8.8% for between-day precision. The method showed potential for being used to monitor forbidden dyes in food constituents. The developed UHPSFC method was compared to the UHPLC-UV method. The orthogonality of Sudan dyes separation by these two methods was demonstrated. Benefits and drawbacks were discussed showing the reliability of both methods for monitoring of studied illegal dyes in real

  17. Determination of Urine Albumin by New Simple High-Performance Liquid Chromatography Method.

    Science.gov (United States)

    Klapkova, Eva; Fortova, Magdalena; Prusa, Richard; Moravcova, Libuse; Kotaska, Karel

    2016-11-01

    A simple high-performance liquid chromatography (HPLC) method was developed for the determination of albumin in patients' urine samples without coeluting proteins and was compared with the immunoturbidimetric determination of albumin. Urine albumin is important biomarker in diabetic patients, but part of it is immuno-nonreactive. Albumin was determined by high-performance liquid chromatography (HPLC), UV detection at 280 nm, Zorbax 300SB-C3 column. Immunoturbidimetric analysis was performed using commercial kit on automatic biochemistry analyzer COBAS INTEGRA ® 400, Roche Diagnostics GmbH, Manheim, Germany. The HLPC method was fully validated. No significant interference with other proteins (transferrin, α-1-acid glycoprotein, α-1-antichymotrypsin, antitrypsin, hemopexin) was found. The results from 301 urine samples were compared with immunochemical determination. We found a statistically significant difference between these methods (P = 0.0001, Mann-Whitney test). New simple HPLC method was developed for the determination of urine albumin without coeluting proteins. Our data indicate that the HPLC method is highly specific and more sensitive than immunoturbidimetry. © 2016 Wiley Periodicals, Inc.

  18. Separation of lanthanum from nuclear fuel solutions by high performance liquid chromatography

    International Nuclear Information System (INIS)

    Lazar, G. C.; Petre, M.; Androne, G.; Benga, A.

    2016-01-01

    This paper presents the separation of uranium, praseodymium and lanthanum from nuclear fuel solutions by high performance liquid chromatography (HPLC). The aim of this study is to establish a minimum concentration of lanthanum which can be analyzed by high performance liquid chromatography, and also to study the effect of uranium concentration on the separation of praseodymium and lanthanum. Optimum gradient mode was established for mixture standard stoc solutions with uranium in a concentration of 1 mg/ml, praseodymium and lanthanum in a concentration range of 1-5 μg/ml from each element. These conditions were applied for the separation of lanthanum from a nuclear fuel solution in which praseodymium and lanthanum were added in a concentration of 3 μg/ml from each element. The elution behavior of lanthanum as a function of the pH and the concentration of the mobile phase, using a mixture of 1-octanesulfonic acid sodium salt with a-hidroxyisobutiric acid is presented. (authors)

  19. Quantitative analysis of benzodiazepines in vitreous humor by high-performance liquid chromatography

    Science.gov (United States)

    Bazmi, Elham; Behnoush, Behnam; Akhgari, Maryam; Bahmanabadi, Leila

    2016-01-01

    Objective: Benzodiazepines are frequently screened drugs in emergency toxicology, drugs of abuse testing, and in forensic cases. As the variations of benzodiazepines concentrations in biological samples during bleeding, postmortem changes, and redistribution could be biasing forensic medicine examinations, hence selecting a suitable sample and a validated accurate method is essential for the quantitative analysis of these main drug categories. The aim of this study was to develop a valid method for the determination of four benzodiazepines (flurazepam, lorazepam, alprazolam, and diazepam) in vitreous humor using liquid–liquid extraction and high-performance liquid chromatography. Methods: Sample preparation was carried out using liquid–liquid extraction with n-hexane: ethyl acetate and subsequent detection by high-performance liquid chromatography method coupled to diode array detector. This method was applied to quantify benzodiazepines in 21 authentic vitreous humor samples. Linear curve for each drug was obtained within the range of 30–3000 ng/mL with coefficient of correlation higher than 0.99. Results: The limit of detection and quantitation were 30 and 100 ng/mL respectively for four drugs. The method showed an appropriate intra- and inter-day precision (coefficient of variation forensic toxicology laboratory. PMID:27635251

  20. Hb A1c Separation by High Performance Liquid Chromatography in Hemoglobinopathies

    Directory of Open Access Journals (Sweden)

    Vani Chandrashekar

    2016-01-01

    Full Text Available Hb A1c measurement is subject to interference by hemoglobin traits and this is dependent on the method used for determination. In this paper we studied the difference between Hb A1c measured by HPLC in hemoglobin traits and normal chromatograms. We also studied the correlation of Hb A1c with age. Hemoglobin analysis was carried out by high performance liquid chromatography. Spearman’s rank correlation was used to study correlation between A1c levels and age. Mann-Whitney U test was used to study the difference in Hb A1c between patients with normal hemoglobin and hemoglobin traits. A total of 431 patients were studied. There was positive correlation with age in patients with normal chromatograms only. No correlation was seen in Hb E trait or beta thalassemia trait. No significant difference in Hb A1c of patients with normal chromatograms and patients with hemoglobin traits was seen. There is no interference by abnormal hemoglobin in the detection of A1c by high performance liquid chromatography. This method cannot be used for detection of A1c in compound heterozygous and homozygous disorders.

  1. EXTRACTION AND QUANTITATIVE ANALYSIS OF ELEMENTAL SULFUR FROM SULFIDE MINERAL SURFACES BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY. (R826189)

    Science.gov (United States)

    A simple method for the quantitative determination of elemental sulfur on oxidized sulfide minerals is described. Extraction of elemental sulfur in perchloroethylene and subsequent analysis with high-performance liquid chromatography were used to ascertain the total elemental ...

  2. Ultra-sensitive high performance liquid chromatography-laser-induced fluorescence based proteomics for clinical applications.

    Science.gov (United States)

    Patil, Ajeetkumar; Bhat, Sujatha; Pai, Keerthilatha M; Rai, Lavanya; Kartha, V B; Chidangil, Santhosh

    2015-09-08

    An ultra-sensitive high performance liquid chromatography-laser induced fluorescence (HPLC-LIF) based technique has been developed by our group at Manipal, for screening, early detection, and staging for various cancers, using protein profiling of clinical samples like, body fluids, cellular specimens, and biopsy-tissue. More than 300 protein profiles of different clinical samples (serum, saliva, cellular samples and tissue homogenates) from volunteers (normal, and different pre-malignant/malignant conditions) were recorded using this set-up. The protein profiles were analyzed using principal component analysis (PCA) to achieve objective detection and classification of malignant, premalignant and healthy conditions with high sensitivity and specificity. The HPLC-LIF protein profiling combined with PCA, as a routine method for screening, diagnosis, and staging of cervical cancer and oral cancer, is discussed in this paper. In recent years, proteomics techniques have advanced tremendously in life sciences and medical sciences for the detection and identification of proteins in body fluids, tissue homogenates and cellular samples to understand biochemical mechanisms leading to different diseases. Some of the methods include techniques like high performance liquid chromatography, 2D-gel electrophoresis, MALDI-TOF-MS, SELDI-TOF-MS, CE-MS and LC-MS techniques. We have developed an ultra-sensitive high performance liquid chromatography-laser induced fluorescence (HPLC-LIF) based technique, for screening, early detection, and staging for various cancers, using protein profiling of clinical samples like, body fluids, cellular specimens, and biopsy-tissue. More than 300 protein profiles of different clinical samples (serum, saliva, cellular samples and tissue homogenates) from healthy and volunteers with different malignant conditions were recorded by using this set-up. The protein profile data were analyzed using principal component analysis (PCA) for objective

  3. Preparation and Characterization of a Polymeric Monolithic Column for Use in High-Performance Liquid Chromatography (HPLC)

    Science.gov (United States)

    Bindis, Michael P.; Bretz, Stacey Lowery; Danielson, Neil D.

    2011-01-01

    The high-performance liquid chromatography (HPLC) experiment, most often done in the undergraduate analytical instrumentation laboratory course, generally illustrates reversed-phase chromatography using a commercial C[subscript]18 silica column. To avoid the expense of periodic column replacement and introduce a choice of columns with different…

  4. tRNA separation by high-performance liquid chromatography using an aggregate of ODS-Hypersil and trioctylmethylammonium chloride

    NARCIS (Netherlands)

    Bischoff, Rainer; Graeser, E.; Mclaughlin, L.W.

    1983-01-01

    High-performance liquid chromatography on a reversed-phase support treated with a tetraalkylammonium salt was used to separate tRNAs from baker's yeast. While resolution by this column appears to result from both anion-exchange and reversed-phase chromatography, it is the hydrophobic interactions

  5. Gas chromatography-mass spectrometry and high-performance liquid chromatographic analyses of thermal degradation products of common plastics

    OpenAIRE

    Pacakova, V.; Leclercq, P.A.

    1991-01-01

    The thermo-oxidation of five commonly used materials, namely low-density polyethylene, retarded polyethylene, paper with a polyethylene foil, a milk package and filled polypropylene, was studied. Capillary gas chromatography and gas chromatography-mass spectrometry were used to analyze the volatile degradation products, while high-performance liquid chromatography was employed to measure polycyclic aromatic hydrocarbons. The results are discussed from the point of view of toxicity of the prod...

  6. Phytochemical Profile of Erythrina variegata by Using High-Performance Liquid Chromatography and Gas Chromatography-Mass Spectroscopy Analyses.

    Science.gov (United States)

    Muthukrishnan, Suriyavathana; Palanisamy, Subha; Subramanian, Senthilkumar; Selvaraj, Sumathi; Mari, Kavitha Rani; Kuppulingam, Ramalingam

    2016-08-01

    Natural products derived from plant sources have been utilized to treat patients with numerous diseases. The phytochemical constituents present in ethanolic leaf extract of Erythrina variegata (ELEV) were identified by using high-performance liquid chromatography (HPLC) and gas chromatography-mass spectroscopy (GC-MS) analyses. Shade dried leaves were powdered and extracted with ethanol for analyses through HPLC to identify selected flavonoids and through GC-MS to identify other molecules. The HPLC analysis of ELEV showed the presence of gallic and caffeic acids as the major components at concentrations of 2.0 ppm and 0.1 ppm, respectively, as well as other components. GC-MS analysis revealed the presence of 3-eicosyne; 3,7,11,15-tetramethyl-2-hexadecen-1-ol; butanoic acid, 3-methyl-3,7-dimethyl-6-octenyl ester; phytol; 1,2-benzenedicarboxylic acid, diundecyl ester; 1-octanol, 2-butyl-; squalene; and 2H-pyran, 2-(7-heptadecynyloxy) tetrahydro-derivative. Because pharmacopuncture is a new evolving natural mode that uses herbal extracts for treating patients with various ailments with minimum pain and maximum effect, the results of this study are particularly important and show that ELEV possesses a wide range of phytochemical constituents, as indicated above, as effective active principle molecules that can be used individually or in combination to treat patients with various diseases. Copyright © 2016. Published by Elsevier B.V.

  7. Evaluation between ultrahigh pressure liquid chromatography and high-performance liquid chromatography analytical methods for characterizing natural dyestuffs.

    Science.gov (United States)

    Serrano, Ana; van Bommel, Maarten; Hallett, Jessica

    2013-11-29

    An evaluation was undertaken of ultrahigh pressure liquid chromatography (UHPLC) in comparison to high-performance liquid chromatography (HPLC) for characterizing natural dyes in cultural heritage objects. A new UHPLC method was optimized by testing several analytical parameters adapted from prior UHPLC studies developed in diverse fields of research. Different gradient elution programs were tested on seven UHPLC columns with different dimensions and stationary phase compositions by applying several mobile phases, flow rates, temperatures, and runtimes. The UHPLC method successfully provided more improved data than that achieved by the HPLC method. Indeed, even though carminic acid has shown circa 146% higher resolution with HPLC, UHPLC resulted in an increase of 41-61% resolution and a decrease of 91-422% limit of detection, depending on the dye compound. The optimized method was subsequently assigned to analyse 59 natural reference materials, in which 85 different components were ascribed with different physicochemical properties, in order to create a spectral database for future characterization of dyes in cultural heritage objects. The majority of these reference samples could be successfully distinguished with one single method through the examination of these compounds' retention times and their spectra acquired with a photodiode array detector. These results demonstrate that UHPLC analyses are extremely valuable for the acquisition of more precise chromatographic information concerning natural dyes with complex mixtures of different and/or closely related physicochemical properties, essential for distinguishing similar species of plants and animals used to colour cultural heritage objects. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. High-performance liquid chromatography of metal complexes of pheophytins a and b

    International Nuclear Information System (INIS)

    Brykina, G.D.; Lazareva, E.E.; Uvarova, M.I.; Shpigun, O.A.

    1997-01-01

    Cu(2), Zn(2), Pb(2), Hg(2), and Ce(4) complexes of phenophytins a and b were synthesized. The chromatographic retention parameters of pheophytins a and b, chlorophylls a and b, and the above complexes were determined under conditions of normal-phase and reversed-phase high-performance liquid chromatography (HPLC). The adsorption of metal pheophytinates in the hexane-n-butanol (96:4)-Silasorb 600 and acetonitrile-ethanol-acetic acid (40:40:16)-Nucleosil C 18 systems was studied by HPLC. Factors that affect the chromatographic and adsorption characteristics of compounds (structural differences between pheophytinates of the a and b series, the nature of the central metal atom, and the nature of the mobile and stationary phases) are discussed. It is demonstrated that pheophytins a and b their metal complexes can be identified and quantiatively determined by HPLC in the concentration range (0.6-44.0)[10 -6 M

  9. ANALISIS RESIDU KLORPIRIFOS DALAM SAYUR-SAYURAN DENGAN TEKNIK HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC

    Directory of Open Access Journals (Sweden)

    Aman Sentosa Panggabean

    2016-06-01

    Full Text Available The research about analysis of chlorpyrifos residue in vegetables by using High Performance Liquid Chromatography (HPLC technique has been done. To obtain the optimal measurement results, the measurement performed several important parameters in the chromatographic system was composition of mobile phase, volume injection sample, flow rate and pH eluent. Optimum measurement conditions obtained was mobile phase composition (water : methanol with 70 : 30, volume injection sample are 5 mL, flow rate are 0.5mL/menit and pH eluent are 7. The analytical performance that obtained is good showed with the reproducibility value as percentage coefficient variance (% CV was 0.0664%, limit of detection (LOD was 0.44 ppm, with a recovery percentage of > 95%. The results obtained showed the HPLC technique can be used for the routine analysis in the determination of chlorpyrifos for the vegetable samples. Keywords: Chlorpyrifos, Vegetables, HPLC.

  10. Quantitative determination of acetaminophen, phenylephrine and carbinoxamine in tablets by high-performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Carina de A. Bastos

    2009-01-01

    Full Text Available An alternative methodology for analysis of acetaminophen (Ace, phenylephrine (Phe and carbinoxamine (Car in tablets by ion-pair reversed phase high performance liquid chromatography was validated. The pharmaceutical preparations were analyzed by using a C18 column (5 μm, 300 mm, 3.9 mm and mobile phase consisting of 60% methanol and 40% potassium monobasic phosphate aqueous solution (62.46 mmol L-1 added with 1 mL phosphoric acid, 0.50 mL triethylamine and 0.25 g sodium lauryl sulfate. Isocratic analysis was performed under direct UV detection at 220 nm for Phe and Car and at 300 nm for Ace within 5 min.

  11. Development of High Performance Liquid Chromatography and Mass Spectrometry: a Key Engine of TCM Modernization

    Directory of Open Access Journals (Sweden)

    Zheng-Xiang Zhang

    2015-04-01

    Full Text Available Traditional Chinese Medicine (TCM has been popular for thousand years in prevention and treatment of chronic diseases synergistically with Western medicine while producing mild healing effects and lower side effects. Although many TCMs have been proven effective by modern pharmacological studies and clinical trials, their bioactive constituents and the remedial mechanisms are still not well understood. Researchers have made great efforts to explore the real theory of TCM for many years with different strategies. Development of high performance liquid chromatography (HPLC and mass spectrometry within recent decade can provide scientists with robust technologies for disclosing the mysterious mask of TCM. In this paper, important innovations of HPLC and mass spectrometry are reviewed in the application of TCM analysis from single compound identification to metabolomic strategy.

  12. A high performance liquid chromatography method for determination of furfural in crude palm oil.

    Science.gov (United States)

    Loi, Chia Chun; Boo, Huey Chern; Mohammed, Abdulkarim Sabo; Ariffin, Abdul Azis

    2011-09-01

    A modified steam distillation method was developed to extract furfural from crude palm oil (CPO). The collected distillates were analysed using high performance liquid chromatography (HPLC) coupled with an ultraviolet diode detector at 284nm. The HPLC method allowed identification and quantification of furfural in CPO. The unique thermal extraction of CPO whereby the fresh fruit bunches (FFB) are first subjected to steam treatment, distinguishes itself from other solvent-extracted or cold-pressed vegetable oils. The presence of furfural was also determined in the fresh palm oil from FFB (without undergoing the normal extraction process), palm olein, palm stearin, olive oil, coconut oil, sunflower oil, soya oil and corn oil. The chromatograms of the extracts were compared to that of standard furfural. Furfural was only detected in CPO. The CPO consignments obtained from four mills were shown to contain 7.54 to 20.60mg/kg furfural. Copyright © 2011 Elsevier Ltd. All rights reserved.

  13. Determination of adenosine phosphates in rat gastrocnemius at various postmortem intervals using high performance liquid chromatography.

    Science.gov (United States)

    Huang, Hong; Yan, Youyi; Zuo, Zhong; Yang, Lin; Li, Bin; Song, Yu; Liao, Linchuan

    2010-09-01

    Although the change in adenosine phosphate levels in muscles may contribute to the development of rigor mortis, the relationship between their levels and the onset and development of rigor mortis has not been well elucidated. In the current study, levels of the adenosine phosphates including adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) in gastrocnemius at various postmortem intervals of 180 rats from different death modes were detected by high performance liquid chromatography. The results showed that the levels of ATP and ADP significantly decreased along with the postmortem period of rats from different death mode whereas the AMP level remained the same. In addition, it was found that changes in the ATP levels in muscles after death correlated well with the development of rigor mortis. Therefore, the ATP level could serve as a reference parameter for the deduction of rigor mortis in forensic science.

  14. Determination of oxymatrine in Sophora Radix by high performance liquid chromatography

    International Nuclear Information System (INIS)

    Yang, Seung Kwon; Yun, Young Ja; Namgung, Mi Ok

    2004-01-01

    A high performance liquid chromatographic method was designed for the quantitative analysis of oxymatrine in Sophora Radix. The separation of oxymatrine was performed by reversed-phase chromatography with a C 18 column and a buffered aqueous solution containing acetonitrile, and monitored by UV absorption at 215 nm. Extraction of oxymatrine in Sophora Radix was carried out using various solvents and extraction methods. The optimum extraction efficiency for the crushed Sophora Radix was achieved by reflux at 80 .deg. C in 50% ethanol for five hours. Most extraction methods used to complicate pretreatments. In this study, sublimation was employed for a extraction method without going through complicate pretreatments. Sublimation was carried out under high vacuum (1x10 -3 torr) and at high temperature (200 .deg. C). Extraction efficiency using Sublimation was found to be inferior to other extraction methods

  15. Cation exchange separation of 16 rare earth metals by microscale high-performance liquid chromatography

    International Nuclear Information System (INIS)

    Ishii, D.; Hirose, A.; Iwasaki, Y.

    1978-01-01

    The separation of rare earth metals has been studied with a microcolumn of 0.5 mm i.d. and 75 mm length, packed with TSK LS-212 high-performance cation exchange resin. A micro-feeder (Model MF-2, from Azumadenki Kogyo) was used to drive carrier and sample solutions through the ion exchange column and detection cell. By combining a 250 μl syringe and a 0.5 mm i.d. sampling tube the micro-feeder, 0.1-1.0 μl rare earth metals were separated within 38 min, using only 304 μl of 0.4M α-hydroxy-isobutyric acid solution adjusted to pH 3.1-6.0 with ammonia solution as gradient carrier solution. The gradient elution was successfully performed by applying a new technique developed for microscale liquid chromatography. (author)

  16. Quantification of sulphur amino acids by ultra-high performance liquid chromatography in aquatic invertebrates.

    Science.gov (United States)

    Thera, Jennifer C; Kidd, Karen A; Dodge-Lynch, M Elaine; Bertolo, Robert F

    2017-12-15

    We examined the performance of an ultra-high performance liquid chromatography method to quantify protein-bound sulphur amino acids in zooplankton. Both cysteic acid and methionine sulfone were linear from 5 to 250 pmol (r 2  = 0.99), with a method detection limit of 13 pmol and 9 pmol, respectively. Although there was no matrix effect on linearity, adjacent peaks and co-eluting noise from the invertebrate proteins increased the detection limits when compared to common standards. Overall, performance characteristics were reproducible and accurate, and provide a means for quantifying sulphur amino acids in aquatic invertebrates, an understudied group. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Core-Shell Columns in High-Performance Liquid Chromatography: Food Analysis Applications

    Science.gov (United States)

    Preti, Raffaella

    2016-01-01

    The increased separation efficiency provided by the new technology of column packed with core-shell particles in high-performance liquid chromatography (HPLC) has resulted in their widespread diffusion in several analytical fields: from pharmaceutical, biological, environmental, and toxicological. The present paper presents their most recent applications in food analysis. Their use has proved to be particularly advantageous for the determination of compounds at trace levels or when a large amount of samples must be analyzed fast using reliable and solvent-saving apparatus. The literature hereby described shows how the outstanding performances provided by core-shell particles column on a traditional HPLC instruments are comparable to those obtained with a costly UHPLC instrumentation, making this novel column a promising key tool in food analysis. PMID:27143972

  18. Identification of glycosaminoglycans using high-performance liquid chromatography on a hydroxyapatite column.

    Science.gov (United States)

    Narita, H; Takeda, Y; Takagaki, K; Nakamura, T; Harata, S; Endo, M

    1995-11-20

    Glycosaminoglycans (heparin, heparan sulfate, dermatan sulfate, chondroitin sulfate, and hyaluronic acid) were labeled with a fluorescent reagent, 2-aminopyridine. The fluoro-labeled glycosaminoglycans were subjected to high-performance liquid chromatography on a hydroxyapatite column. The binding property of each glycosaminoglycan to hydroxyapatite was different. The structural properties of glycosaminoglycans bound to hydroxyapatite were then investigated using chemical desulfated or enzymic depolymerized glycosaminoglycans. This revealed that the sulfate content and molecular weight of the glycosaminoglycans correlated with their binding properties to hydroxyapatite. Desulfated dermatan sulfate but not desulfated chondroitin 6-sulfate bound to the hydroxyapatite. These data indicate that iduronic acid residues of glycosaminoglycans are important for the binding property. The method described which uses hydroxyapatite columns facilitates rapid separation and microanalysis of the glycosaminoglycans, especially dermatan sulfate and chondroitin sulfate.

  19. Burn-up measurements on nuclear reactor fuels using high performance liquid chromatography

    International Nuclear Information System (INIS)

    Sivaraman, N.; Subramaniam, S.; Srinivasan, T.G.; Vasudeva Rao, P.R.

    2002-01-01

    Burn-up measurements on thermal as well as fast reactor fuels were carried out using high performance liquid chromatography (HPLC). A column chromatographic technique using di-(2-ethylhexyl) phosphoric acid (HDEHP) coated column was employed for the isolation of lanthanides from uranium, plutonium and other fission products. Ion-pair HPLC was used for the separation of individual lanthanides. The atom percent fissions were calculated from the concentrations of the lanthanide (neodymium in the case of thermal reactor and lanthanum for the fast reactor fuels) and from uranium and plutonium contents of the dissolver solutions. The HPLC method was also used for determining the fractional fissions from uranium and plutonium for the thermal reactor fuel. (author)

  20. High-performance liquid chromatography for determination of α-tocopherol in vegetables

    Directory of Open Access Journals (Sweden)

    Marcin Horbowicz

    2013-12-01

    Full Text Available A simple method for the determination of α-tocopherol in vegetables is described. The procedure consists of the following steps: saponification, extraction, silica-column clean-up, and high-performance liquid chromatography. Elution time for D, L-α-tocopherol was 9.0 min using a Zorbax Sil (250 x 4.6 mm column and an isocratic mobile phase of hexane-methanol (99.3 + 0.7, with a flow rate of 1 ml/min, and detection at 292 nm using a variable UV detector. The average recovery of α-tocopherol was 91.2%, and the minimum detectable amount was 0.1 mg/100 g of fresh vegetable tissue. This method is comparable to gas-chromatographic determination of α-tocopherol, but has fewer analytical steps and gives more reproducible results.

  1. Determination of carbohydrates by high performance anion chromatography-pulsed amperometric detection in mushrooms.

    Science.gov (United States)

    Zhou, Shuai; Tang, Qingjiu; Luo, Xi; Xue, Jun-Jie; Liu, Yanfang; Yang, Yan; Zhang, Jingsong; Feng, Na

    2012-01-01

    A method of detecting carbohydrates (fucose, trehalose, mannitol, arabitol, mannose, glucose, galactose, fructose, and ribose) by high-performance anion chromatography-pulsed amperometric detection (HAPEC-PAD) was established. The conditions are: CarboPac MA1 column, NaOH as the eluent, temperature 30°C, Au working electrode, Ag/AgCl reference electrode, and flow rate 0.4 mL/min. These nine analytes, which yielded high resolution by this method, could be detected in 40 minutes. Mushrooms were tested and good precision, stability, and reproducibility were achieved. This method is suitable for mushroom samples and could support research and development on sugar and sugar alcohol, which contains special effects.

  2. SIMULTANEOUS DETERMINATION OF PARACETAMOL AND IBUPROFENE MIXTURES BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

    Directory of Open Access Journals (Sweden)

    Sophi Damayanti

    2010-06-01

    Full Text Available Analytical method for the determination of paracetamol and ibuprofene mixtures has been developed by High Performance Liquid Chromatography using C-18 column and acetinitrile - phosphate buffer pH = 4.5 (75:25 containing 0.075% sodium hexanesulfunate as a mobile phase. The detector was set at 215 nm. Using such conditions, retention time for paracetamol and ibuprofen was 4.89 and 7.11 min, respectively. The recovery for paracetamol and ibuprofen was found to be 101.07± 0.73% and 102.02 ± 1.58%, respectively. The detector limits of the method was 1.30 and 1.60 μg/mL with the relative standard deviation (RSD 0.74 and 1.52% for paracetamol and ibuprofen, respectively.   Keywords: paracetamol, ibuprofen, multi-component, validation, HPLC.

  3. Determination of phenmedipham and desmedipham in a commercial herbicide by high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Lončar Eva S.

    2004-01-01

    Full Text Available Betanal AM-11 is the herbicide that is using to control one-year old weeds with wide leafs in sugar beet fields. Active ingredients of the herbicide are phenmedipham and desmedipham. Commercial emulsifiable concentrate (EC contains 80 g/L ( 10% of each active compound. We applied high performance liquid chromatography (HPLC with diode array detector (DAD at 254 nm for the determination of phenmedipham and desmedipham in commercial samples of Betanal AM-11. This method involves reversed-phase separation of the components on C-18 bonded silica with methanol-water (51+49, v/v as the eluent. The procedure was highly selective and reproducible and can be successfully used in determining contents of phenmedipham and desmedipham on micro and macro levels.

  4. Determination of free urinary cortisol in cushing's syndrome using reversed-phase high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Eduardo Kinio Sugawara

    2010-01-01

    Full Text Available Determination of free urinary cortisol is a test of choice in the diagnosis of Cushing's syndrome. In this study, cortisol was quantified using reversed-phase high-performance liquid chromatography (RP-HPLC in urine samples previously extracted with ether and using triamcinolone acetonide as internal standard (IS. A BDS-Hypersil-C18® column, water-acetonitrile (72:28; v/v, with a flow rate of 1.0 mL/min and detection at 243 nm were used. This method showed to be both effective and efficient, with sensitivity and linearity ranging from 2.50 to 150 μg/L, and can be used in substitution to the radioimmunoassay technique within this concentration range.

  5. A simple high performance liquid chromatography method for analyzing paraquat in soil solution samples.

    Science.gov (United States)

    Ouyang, Ying; Mansell, Robert S; Nkedi-Kizza, Peter

    2004-01-01

    A high performance liquid chromatography (HPLC) method with UV detection was developed to analyze paraquat (1,1'-dimethyl-4,4'-dipyridinium dichloride) herbicide content in soil solution samples. The analytical method was compared with the liquid scintillation counting (LSC) method using 14C-paraquat. Agreement obtained between the two methods was reasonable. However, the detection limit for paraquat analysis was 0.5 mg L(-1) by the HPLC method and 0.05 mg L(-1) by the LSC method. The LSC method was, therefore, 10 times more precise than the HPLC method for solution concentrations less than 1 mg L(-1). In spite of the high detection limit, the UC (nonradioactive) HPLC method provides an inexpensive and environmentally safe means for determining paraquat concentration in soil solution compared with the 14C-LSC method.

  6. Development of high performance liquid chromatography for rapid determination of burn-up of nuclear fuels

    International Nuclear Information System (INIS)

    Joseph, M.; Karunasagar, D.; Saha, B.

    1996-01-01

    Burn-up an important parameter during evaluation of the performance of any nuclear fuel. Among the various techniques available, the preferred one for its determination is based on accurate measurement of a suitable fission product monitor and the residual heavy elements. Since isotopes of rare earth elements are generally used as burn-up monitors, conditions were standardized for rapid separation (within 15 minutes) of light rare earths using high performance liquid chromatography based on either anion exchange (Partisil 10 SAX) in methanol-nitric acid medium or by cation exchange on a reverse phase column (Spherisorb 5-ODS-2 or Supelcosil LC-18) dynamically modified with 1-octane sulfonate or camphor-10-sulfonic acid (β). Both these methods were assessed for separation of individual fission product rare earths from their mixtures. A new approach has been examined in detail for rapid assay of neodymium, which appears promising for faster and accurate measurement of burn-up. (author)

  7. High performance liquid chromatography method for the determination of cinnamyl alcohol dehydrogenase activity in soybean roots.

    Science.gov (United States)

    dos Santos, W D; Ferrarese, Maria de Lourdes Lucio; Ferrarese-Filho, O

    2006-01-01

    This study proposes a simple, quick and reliable method for determining the cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.195) activity in soybean (Glycine max L. Merr.) roots using reversed-phase high performance liquid chromatography (RP-HPLC). The method includes a single extraction of the tissue and conduction of the enzymatic reaction at 30 degrees C with cinnamaldehydes (coniferyl or sinapyl), substrates of CAD. Disappearance of the substrates in the reaction mixture is monitored at 340 nm (for coniferaldehyde) or 345 nm (for sinapaldehyde) by isocratic elution with methanol/acetic acid through a GLC-ODS (M) column. This HPLC technique furnishes a rapid and reliable measure of cinnamaldehyde substrates, and may be used as an alternative tool to analyze CAD activity in enzyme preparation without previous purification.

  8. Discrimination of Black Ball-point Pen Inks by High Performance Liquid Chromatography (HPLC)

    International Nuclear Information System (INIS)

    Mohamed Izzharif Abdul Halim; Norashikin Saim; Rozita Osman; Halila Jasmani; Nurul Nadhirah Zainal Abidin

    2013-01-01

    In this study, thirteen types of black ball-point pen inks of three major brands were analyzed using high performance liquid chromatography (HPLC). Separation of the ink components was achieved using Bondapak C-18 column with gradient elution using water, ethanol and ethyl acetate. The chromatographic data obtained at wavelength 254.8 nm was analyzed using agglomerative hierarchical clustering (AHC) and principle component analysis (PCA). AHC was able to group the inks into three clusters. This result was supported by PCA, whereby distinct separation of the three different brands was achieved. Therefore, HPLC in combination with chemometric methods may be a valuable tool for the analysis of black ball-point pen inks for forensic purposes. (author)

  9. [Detection of the preservative chlorphenesin in cosmetics by high-performance liquid chromatography].

    Science.gov (United States)

    Ikarashi, Yoshiaki; Miyazawa, Norimasa; Shimamura, Kimio; Sato, Nobuo; Yoshizawa, Ken-ichi; Hayashi, Masahito; Takano, Katsuhiro; Miyamoto, Michiko; Kojima, Takashi; Sakaguchi, Hiroshi; Fujiio, Makiko

    2009-01-01

    A simple determination method for preservative chlorphenesin in cosmetics was developed. Cosmetic samples were dissolved in methanol. The sample solution was analyzed by high-performance liquid chromatography (HPLC) with ODS column, using water-methanol (55:45) or water-acetonitrile (3:1) adjusted to pH 2.5 with phosphoric acid as the mobile phase. Chlorphenesin was detected with ultraviolet light detection at 280 nm. A linear relation was obtained between the peak areas and the concentrations of chlorphenesin in the range of 1-500 microg/ml. The determination limit of chlorphenesin was 1-2 microg/ml. Recoveries of chlorphenesin spiked in lotion and milky lotion at the levels of 0.03% and 0.3% were 98.8-100.0%. This method was applied for cosmetics including 0.03% and 0.3% of chlorphenesin and their content corresponded with the determined values.

  10. [Analysis of the preservative chlorphenesin in cosmetics by high performance liquid chromatography].

    Science.gov (United States)

    Zhu, Huijuan; Zhang, Weiqiang; Yang, Yanwei; Zhu, Ying

    2014-01-01

    An analytical method was developed for the determination of the preservative of chlorphenesin in cosmetics by high performance liquid chromatography (HPLC). A C18 column (250 mm x 4.6 mm, 5 microm) and a photodiode array detector were used. The mobile phase was methanol-water (55:45, v/v) with a flow rate of 1.0 mL/min. The detection wavelength was set at 280 nm and the column temperature was 25 degrees C. The limit of detection was 3 ng. A good linear relationship was obtained between the peak area and the mass concentration of chlorphenesin in the range of 1 - 500 mg/L and the correlation coefficient was 1.000 0. The recoveries of chlorphenesin at different spiked levels were 99.0% - 103% with the relative standard deviations (RSD) chlorphenesin in cosmetics.

  11. [Determination of aspirin and free salicylic acid in lysinipirine injection by high performance liquid chromatography].

    Science.gov (United States)

    Dong, Yu; Zhao, Yuan-zheng; Zhang, Yi-na

    2002-05-01

    The contents of aspirin and free salicylic acid in lysinipirine injection were determined by high performance liquid chromatography (HPLC). A Hypersil BDS C18 column was used with the mobile phase of methanol-water-acetic acid (35:65:3, volume ratio) and the detection wavelength of 280 nm. The average recoveries of aspirin and salicylic acid added were 99.27% (RSD = 0.8%) and 99.61%(RSD = 1.3%), respectively. The calibration curves had good linearity in the range of 0.028 g/L -0.141 mg/L and 0.77 mg/L -3.85 mg/L, and the correlation coefficients were 0.9999 and 0.9998 for aspirin and salicylic acid respectively.

  12. [Determination of glycyrrhizinic acid in biotransformation system by reversed-phase high performance liquid chromatography].

    Science.gov (United States)

    Li, Hui; Lu, Dingqiang; Liu, Weimin

    2004-05-01

    A method for determining glycyrrhizinic acid in the biotransformation system by reversed-phase high performance liquid chromatography (RP-HPLC) was developed. The HPLC conditions were as follows: Hypersil C18 column (4.6 mm i.d. x 250 mm, 5 microm) with a mixture of methanol-water-acetic acid (70:30:1, v/v) as the mobile phase; flow rate at 1.0 mL/min; and UV detection at 254 nm. The linear range of glycyrrhizinic acid was 0.2-20 microg. The recoveries were 98%-103% with relative standard deviations between 0.16% and 1.58% (n = 3). The method is simple, rapid and accurate for determining glycyrrhizinic acid.

  13. Measurement of food flavonoids by high-performance liquid chromatography: A review.

    Science.gov (United States)

    Merken, H M; Beecher, G R

    2000-03-01

    The flavonoids are plant polyphenols found frequently in fruits, vegetables, and grains. Divided into several subclasses, they include the anthocyanidins, pigments chiefly responsible for the red and blue colors in fruits, fruit juices, wines, and flowers; the catechins, concentrated in tea; the flavanones and flavanone glycosides, found in citrus and honey; and the flavones, flavonols, and flavonol glycosides, found in tea, fruits, vegetables, and honey. Known for their hydrogen-donating antioxidant activity as well as their ability to complex divalent transition metal cations, flavonoids are propitious to human health. Computer-controlled high-performance liquid chromatography (HPLC) has become the analytical method of choice. Many systems have been developed for the detection and quantification of flavonoids across one, two, or three subclasses. A summary of the various HPLC and sample preparation methods that have been employed to quantify individual flavonoids within a subclass or across several subclasses are tabulated in this review.

  14. Development of high performance liquid chromatography method for miconazole analysis in powder sample

    Science.gov (United States)

    Hermawan, D.; Suwandri; Sulaeman, U.; Istiqomah, A.; Aboul-Enein, H. Y.

    2017-02-01

    A simple high performance liquid chromatography (HPLC) method has been developed in this study for the analysis of miconazole, an antifungal drug, in powder sample. The optimized HPLC system using C8 column was achieved using mobile phase composition containing methanol:water (85:15, v/v), a flow rate of 0.8 mL/min, and UV detection at 220 nm. The calibration graph was linear in the range from 10 to 50 mg/L with r 2 of 0.9983. The limit of detection (LOD) and limit of quantitation (LOQ) obtained were 2.24 mg/L and 7.47 mg/L, respectively. The present HPLC method is applicable for the determination of miconazole in the powder sample with a recovery of 101.28 % (RSD = 0.96%, n = 3). The developed HPLC method provides short analysis time, high reproducibility and high sensitivity.

  15. Determination of 6-mercaptopurine and azathioprine in plasma by high-performance liquid chromatography.

    Science.gov (United States)

    Ding, T L; Benet, L Z

    1979-07-21

    Using 1-ml plasma samples, levels of 6-mercaptopurine (6MP) as low as 5 ng/ml and azathioprine (AZA) as low as 40 ng/ml can be detected using a high-performance liquid chromatography reversed-phase column procedure following extraction. Both compounds were stable in frozen plasma for seven weeks. AZA stability in blood was temperature dependent; the half-lives of AZA breakdown to 6MP at 37 degrees were 28 and 46 min in blood drawn from two rhesus monkeys. Plasma levels of 6MP were measured in a rhesus monkey following 6MP (1.47 mg/kg) and AZA (3 mg/kg) intravenous administration. 6MP levels were also measured in three renal transplant patients on daily 50- and 100-mg AZA doses. Peak levels (45-75 ng/ml) were reached within an hour and 6MP levels were detected for up to 7 h.

  16. Determination of 15 isoflavone isomers in soy foods and supplements by high-performance liquid chromatography.

    Science.gov (United States)

    Yanaka, Kaoru; Takebayashi, Jun; Matsumoto, Teruki; Ishimi, Yoshiko

    2012-04-25

    Soy isoflavone is the generic name for the isoflavones found in soy. We determined the concentrations of 15 soy isoflavone species, including 3 succinyl glucosides, in 22 soy foods and isoflavone supplements by high-performance liquid chromatography (HPLC). The total isoflavone contents in 14 soy foods and 8 supplements ranged from 45 to 735 μg/g and from 1,304 to 90,224 μg/g, respectively. Higher amounts of succinyl glucosides were detected in natto, a typical fermented soy product in Japan; these ranged from 30 to 80 μg/g and comprised 4.1-10.9% of the total isoflavone content. In soy powder, 59 μg/g of succinyl glucosides were detected, equivalent to 4.6% of the total isoflavone content. These data suggest that the total isoflavone contents may be underestimated in the previous studies that have not included succinyl glucosides, especially for Bacillus subtilis -fermented soy food products.

  17. Quality control of 99mTc-DTPA-octreotide by reverse high performance liquid chromatography

    International Nuclear Information System (INIS)

    Cheng, Z.; Lin, Q.F.; Jin, X.H.; Wang, F.; Bai, H.S.; Chen, D.M.; Fan, H.Q.; Du, J.

    1998-01-01

    DTPA-Octreotide(Pentetreotide), a somatostatin analogue which can bind specifically and with high affinity to somatostatin receptor in vitro and vivo, labeled with 99m Tc by tin reduction in acetate buffer, has been characterized by Reverse-phase High performance Liquid Chromatography. The effect of different solvents, mobile phase pH, linear gradient and the injected volume on the separation efficiency was evaluated. The results show that the separation efficiency is best using μBondapak-C 18 (300x3.9 mm 2 ), linear gradient of 40% to 80% methanol (1.0 ml/min) in 0.05M acetate buffer (pH 5.5) over a 30 min period and maintaining for another 10 min. The labeled product is a mixture which mainly consists of five components (a, b, c, d, e) successfully proved by HPLC. Paper chromatography is also evaluated in this paper. It may be used to determine the radiochemical purity of the labeling product, but is not a good choice for the verification each components. (author)

  18. Neutral monosaccharide composition analysis of plant-derived oligo- and polysaccharides by high performance liquid chromatography.

    Science.gov (United States)

    Yan, Jun; Shi, Songshan; Wang, Hongwei; Liu, Ruimin; Li, Ning; Chen, Yonglin; Wang, Shunchun

    2016-01-20

    A novel analytical method for neutral monosaccharide composition analysis of plant-derived oligo- and polysaccharides was developed using hydrophilic interaction liquid chromatography coupled to a charged aerosol detector. The effects of column type, additives, pH and column temperature on retention and separation were evaluated. Additionally, the method could distinguish potential impurities in samples, including chloride, sulfate and sodium, from sugars. The results of validation demonstrated that this method had good linearity (R(2) ≥ 0.9981), high precision (relative standard deviation ≤ 4.43%), and adequate accuracy (94.02-103.37% recovery) and sensitivity (detection limit: 15-40 ng). Finally, the monosaccharide compositions of the polysaccharide from Eclipta prostrasta L. and stachyose were successfully profiled through this method. This report represents the first time that all of these common monosaccharides could be well-separated and determined simultaneously by high performance liquid chromatography without additional derivatization. This newly developed method is convenient, efficient and reliable for monosaccharide analysis. Copyright © 2015 Elsevier Ltd. All rights reserved.

  19. Improved separation of conjugated fatty acid methyl esters by silver ion-high-performance liquid chromatography.

    Science.gov (United States)

    Sehat, N; Rickert, R; Mossoba, M M; Kramer, J K; Yurawecz, M P; Roach, J A; Adlof, R O; Morehouse, K M; Fritsche, J; Eulitz, K D; Steinhart, H; Ku, Y

    1999-04-01

    Operating from one to six silver ion-high-performance liquid chromatography (Ag+-HPLC) columns in series progressively improved the resolution of the methyl esters of conjugated linoleic acid (CLA) isomeric mixtures from natural and commercial products. In natural products, the 8 trans, 10 cis-octadecadienoic (18:2) acid was resolved from the more abundant 7 trans, 9 cis-18:2, and the 10 trans, 12 cis-18:2 was separated from the major 9 cis, 11 trans-18:2 peak. In addition, both 11 trans, 13 cis-18:2 and 11 cis, 13 trans-18:2 isomers were found in natural products and were separated; the presence of the latter, 11 cis, 13 trans-18:2, was established in commercial CLA preparations. Three Ag+-HPLC columns in series appeared to be the best compromise to obtain satisfactory resolution of most CLA isomers found in natural products. A single Ag+-HPLC column in series with one of several normal-phase columns did not improve the resolution of CLA isomers as compared to that of the former alone. The 20:2 conjugated fatty acid isomers 11 cis, 13 trans-20:2 and 12 trans, 14 cis-20:2, which were synthesized by alkali isomerization from 11 cis, 14 cis-20:2, eluted in the same region of the Ag+-HPLC chromatogram just before the corresponding geometric CLA isomers. Therefore, CLA isomers will require isolation based on chain length prior to Ag+-HPLC separation. The positions of conjugated double bonds in 20:2 and 18:2 isomers were established by gas chromatography-electron ionization mass spectrometry as their 4,4-dimethyloxazoline derivatives. The double-bond geometry was determined by gas chromatography-direct deposition-Fourier transform infrared spectroscopy and by the Ag+-HPLC relative elution order.

  20. Screening of Norharmane from Seven Cyanobacteria by High-performance Liquid Chromatography.

    Science.gov (United States)

    Karan, Tunay; Erenler, Ramazan

    2017-10-01

    Cyanobacteria, including pharmaceutically and medicinally valuable compounds attract the great attention lately. Norharmane (9H-pyrido (3,4-b) indole found in some cyanobacteria revealed a great number of biological effects. Seven cyanobacteria were isolated and identified from Yesilirmak River and Gaziosmanpasa University Campus to determine the norharmane content. Cyanobacteria collected from Tokat, Turkey were isolated and identified by morphologically. Norharmane (9H-pyrido [3,4-b] indole) quantities were presented for seven cyanobacteria, Chroococcus minutus (Kütz.) Nägeli, Geitlerinema carotinosum (Geitler) Anagnostidis, Nostoc linckia Bornet ex Bornet and Flahault, Anabaena oryzae F. E. Fritsch, Oscillatoria limnetica Lemmermann, Phormidium sp . Kützing ex Gomont, and Cylindrospermum sp . Kutzing ex E. Bornet and C. Flahault by high-performance liquid chromatography. The norharmane amount indicated for cyanobacterial culture media altered in a species-dependent kind in the range of 0.81-10.87 μg/g. C. minutus produced the most norharmane among the investigated cyanobacteria as 10.87 μg/g. Cyanobacteria could be an important source of norharmane as well as pharmaceutically valuable compounds. Seven cyanobacteria were isolated and identified from Yesilirmak RiverQuantitative analysis of norharmane was executed on isolated cyanobacteriaFour cyanobecteria species included the norharmane Chroococcus minutus contained the most norharmane (10.87 μg/g). Abbreviations used: HPLC: High performance liquid chromatograph.

  1. Screening of Norharmane from Seven Cyanobacteria by High-performance Liquid Chromatography

    Science.gov (United States)

    Karan, Tunay; Erenler, Ramazan

    2017-01-01

    Background: Cyanobacteria, including pharmaceutically and medicinally valuable compounds attract the great attention lately. Norharmane (9H-pyrido (3,4-b) indole found in some cyanobacteria revealed a great number of biological effects. Objective: Seven cyanobacteria were isolated and identified from Yesilirmak River and Gaziosmanpasa University Campus to determine the norharmane content. Materials and Methods: Cyanobacteria collected from Tokat, Turkey were isolated and identified by morphologically. Norharmane (9H-pyrido [3,4-b] indole) quantities were presented for seven cyanobacteria, Chroococcus minutus (Kütz.) Nägeli, Geitlerinema carotinosum (Geitler) Anagnostidis, Nostoc linckia Bornet ex Bornet and Flahault, Anabaena oryzae F. E. Fritsch, Oscillatoria limnetica Lemmermann, Phormidium sp. Kützing ex Gomont, and Cylindrospermum sp. Kutzing ex E. Bornet and C. Flahault by high-performance liquid chromatography. Results: The norharmane amount indicated for cyanobacterial culture media altered in a species-dependent kind in the range of 0.81–10.87 μg/g. C. minutus produced the most norharmane among the investigated cyanobacteria as 10.87 μg/g. Conclusion: Cyanobacteria could be an important source of norharmane as well as pharmaceutically valuable compounds. SUMMARY Seven cyanobacteria were isolated and identified from Yesilirmak RiverQuantitative analysis of norharmane was executed on isolated cyanobacteriaFour cyanobecteria species included the norharmaneChroococcus minutus contained the most norharmane (10.87 μg/g). Abbreviations used: HPLC: High performance liquid chromatograph. PMID:29142439

  2. An optimized method for the measurement of acetaldehyde by high-performance liquid chromatography.

    Science.gov (United States)

    Guan, Xiangying; Rubin, Emanuel; Anni, Helen

    2012-03-01

    Acetaldehyde is produced during ethanol metabolism predominantly in the liver by alcohol dehydrogenase and rapidly eliminated by oxidation to acetate via aldehyde dehydrogenase. Assessment of circulating acetaldehyde levels in biological matrices is performed by headspace gas chromatography and reverse phase high-performance liquid chromatography (RP-HPLC). We have developed an optimized method for the measurement of acetaldehyde by RP-HPLC in hepatoma cell culture medium, blood, and plasma. After sample deproteinization, acetaldehyde was derivatized with 2,4-dinitrophenylhydrazine (DNPH). The reaction was optimized for pH, amount of derivatization reagent, time, and temperature. Extraction methods of the acetaldehyde-hydrazone (AcH-DNP) stable derivative and product stability studies were carried out. Acetaldehyde was identified by its retention time in comparison with AcH-DNP standard, using a new chromatography gradient program, and quantitated based on external reference standards and standard addition calibration curves in the presence and absence of ethanol. Derivatization of acetaldehyde was performed at pH 4.0 with an 80-fold molar excess of DNPH. The reaction was completed in 40 minutes at ambient temperature, and the product was stable for 2 days. A clear separation of AcH-DNP from DNPH was obtained with a new 11-minute chromatography program. Acetaldehyde detection was linear up to 80 μM. The recovery of acetaldehyde was >88% in culture media and >78% in plasma. We quantitatively determined the ethanol-derived acetaldehyde in hepatoma cells, rat blood and plasma with a detection limit around 3 μM. The accuracy of the method was volume (70 μl) plasma sampling. An optimized method for the quantitative determination of acetaldehyde in biological systems was developed using derivatization with DNPH, followed by a short RP-HPLC separation of AcH-DNP. The method has an extended linear range, is reproducible and applicable to small-volume sampling of culture

  3. High performance liquid chromatography--atomic fluorescence spectrometric determination of arsenic species in beer samples

    International Nuclear Information System (INIS)

    Melo Coelho, N.M.; Parrilla, Carmen; Cervera, M.L.; Pastor, A.; Guardia, M. de la

    2003-01-01

    A method has been developed for the direct determination of As(III), dimethylarsinic acid (DMA), monomethylarsonic acid (MMA) and As(V) in beers by hydride generation--atomic fluorescence spectrometry after separation of arsenic species by high performance liquid chromatography. Compounds were separated by anion-exchange chromatography with isocratic elution using KH 2 PO 4 /K 2 HPO 4 as mobile phase with elution times of 1.67, 2.08, 6.52 and 10.72 min for As(III), DMA, MMA and As(V), respectively. Parameters affecting the hydride generation of all arsenic species were studied and the best conditions were established as a reaction coil of 150 cm, for a sample injected volume of 100 μl, a 4.0% (m/v) solution of sodium tetrahydroborate and 2.0 mol l -1 hydrochloric acid with flow rates of 2.7 and 1.7 ml min -1 , respectively and a flow rate of 500 ml min -1 for the argon carrier gas. Under the best experimental conditions, the detection limit was found to be 0.12, 0.20, 0.27 and 0.39 μg l -1 for As(III), DMA, MMA and As(V), respectively. The relative standard deviation for eight independent determinations varied from 3.9 till 8.9% for species considered at a concentration level of 10.0 μg l -1 . Recovery and comparative studies evidenced that the method is suitable for the accurate determination of arsenic species in water and beer samples

  4. Determination of aflatoxins in medicinal plants by high-performance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Siddique, Nadeem A; Mujeeb, Mohd; Ahmad, Sayeed; Panda, Bibhu P; Makhmoor, Mohd

    2013-01-01

    The intention of the proposed work is to study the presence of the aflatoxins B1, B2, G1 and G2 in medicinal plants, namely Mucuna pruriens, Delphinium denudatum and Portulaca oleraceae. The aflatoxins were extracted, purified by immunoaffinity column chromatography and analysed by high-performance liquid chromatography-tandem quadrupole mass spectrometry with electrospray ionisation (HPLC-MS/MS). Fungal count was carried out in PDA media. A good linear relationship was found for AFB1, AFB2, AFG1 and AFG2 at 1-10 ppb (r>0.9995). The analyte accuracy under three different spiking levels was 86.7-108.1 %, with low per cent relative standard deviations in each case. The aflatoxins can be separated within 5 to7 min using an Agilent XDB C18-column. We found that AFB1 and AFB2 were in trace amounts below the detection limit in M. pruriens whilst they were not detected in D. denudatum. P. oleraceae was found to be contaminated with AFB1 and AFB2. AFG1 and AFG2 were not detected in M. pruriens, P. oleraceae and were below the detection limit in D. denudatum. This was consistent with very low numbers of fungal colonies observed after 6 hr of incubation. The analytical method developed is simple, precise, accurate, economical and can be effectively used to determine the aflatoxins in medicinal plants and therefore to control the quality of products. The aflatoxin levels in the plant extracts examined were related to the minimal fungal load in the medicinal plants examined.

  5. High performance liquid chromatography--atomic fluorescence spectrometric determination of arsenic species in beer samples

    Energy Technology Data Exchange (ETDEWEB)

    Melo Coelho, N.M.; Parrilla, Carmen; Cervera, M.L.; Pastor, A.; Guardia, M. de la

    2003-04-10

    A method has been developed for the direct determination of As(III), dimethylarsinic acid (DMA), monomethylarsonic acid (MMA) and As(V) in beers by hydride generation--atomic fluorescence spectrometry after separation of arsenic species by high performance liquid chromatography. Compounds were separated by anion-exchange chromatography with isocratic elution using KH{sub 2}PO{sub 4}/K{sub 2}HPO{sub 4} as mobile phase with elution times of 1.67, 2.08, 6.52 and 10.72 min for As(III), DMA, MMA and As(V), respectively. Parameters affecting the hydride generation of all arsenic species were studied and the best conditions were established as a reaction coil of 150 cm, for a sample injected volume of 100 {mu}l, a 4.0% (m/v) solution of sodium tetrahydroborate and 2.0 mol l{sup -1} hydrochloric acid with flow rates of 2.7 and 1.7 ml min{sup -1}, respectively and a flow rate of 500 ml min{sup -1} for the argon carrier gas. Under the best experimental conditions, the detection limit was found to be 0.12, 0.20, 0.27 and 0.39 {mu}g l{sup -1} for As(III), DMA, MMA and As(V), respectively. The relative standard deviation for eight independent determinations varied from 3.9 till 8.9% for species considered at a concentration level of 10.0 {mu}g l{sup -1}. Recovery and comparative studies evidenced that the method is suitable for the accurate determination of arsenic species in water and beer samples.

  6. Multi-Instrument Characterization of the Surfaces and Materials in Microfabricated, Carbon Nanotube-Templated Thin Layer Chromatography Plates. An Analogy to ‘The Blind Men and the Elephant’

    Energy Technology Data Exchange (ETDEWEB)

    Jensen, David S.; Kanyal, Supriya S.; Madaan, Nitesh; Hancock, Jared M.; Dadson, Andrew; Vail, Michael A.; Vanfleet, Richard; Shutthanandan, V.; Zhu, Zihua; Engelhard, Mark H.; Linford, Matthew R.

    2013-08-08

    Herein we apply a suite of surface/materials analytical tools to characterize some of the materials created in the production of microfabricated thin layer chromatography plates. Techniques used include X-ray photoelectron spectroscopy (XPS), valence band spectroscopy, static time-of-flight secondary ion spectrometry (ToF-SIMS) in both positive and negative ion modes, Rutherford backscattering spectroscopy (RBS), and helium ion microscopy (HIM). Materials characterized include: the Si(100) substrate with native oxide: Si/SiO2, alumina (35 nm) deposited as a diffusion barrier on the Si/SiO2: Si/SiO2/Al2O3, iron (6 nm) thermally evaporated on the Al2O3: Si/SiO2/Al2O3/Fe, the iron film annealed in H2 to make Fe catalyst nanoparticles: Si/SiO2/Al2O3/Fe(NP), and carbon nanotubes (CNTs) grown from the Fe nanoparticles: Si/SiO2/Al2O3/Fe(NP)/CNT. The Fe thin films and nanoparticles are found in an oxidized state. Some of the analyses of the CNTs/CNT forests reported appear to be unique: the CNT forest appears to exhibit an interesting ‘channeling’ phenomenon by RBS, we observe an odd-even effect in the ToF-SIMS spectra of Cn- species for n = 1 – 6, with ions at even n showing greater intensity than the neighboring signals, and ions with n ≥ 6 showing a steady decrease in intensity, and valence band characterization of CNTs using X-radiation is reported. The information obtained from the combination of the different analytical tools provides a more complete understanding of our materials than a single technique, which is analogous to the story of ‘The Blind Men and the Elephant’. (Of course there is increasing emphasis on the use of multiple characterization tools in surface and materials analysis.) The raw XPS and ToF-SIMS spectra from this study will be submitted to Surface Science Spectra for archiving.

  7. Plasma capric acid concentrations in healthy subjects determined by high-performance liquid chromatography.

    Science.gov (United States)

    Shrestha, Rojeet; Hui, Shu-Ping; Imai, Hiromitsu; Hashimoto, Satoru; Uemura, Naoto; Takeda, Seiji; Fuda, Hirotoshi; Suzuki, Akira; Yamaguchi, Satoshi; Hirano, Ken-Ichi; Chiba, Hitoshi

    2015-09-01

    Capric acid (FA10:0, decanoic acid) is a medium-chain fatty acid abundant in tropical oils such as coconut oil, whereas small amounts are present in milk of goat, cow, and human. Orally ingested FA10:0 is transported to the liver and quickly burnt within it. Only few reports are available for FA10:0 concentrations in human plasma. Fasting (n = 5, male/female = 3/2, age 31 ± 9.3 years old) and non-fasting (n = 106, male/female = 44/62, age 21.9 ± 3.2 years old) blood samples were collected from apparently healthy Japanese volunteers. The total FA10:0 in the plasma were measured by high-performance liquid chromatography after derivatization with 2-nitrophenylhydrazine followed by UV detection. Inter and intra-assay coefficient of variation of FA10:0 assay at three different concentrations ranged in 1.7-3.9 and 1.3-5.4%, respectively, with an analytical recovery of 95.2-104.0%. FA10:0 concentration was below detection limit (0.1 µmol/L) in each fasting human plasma. FA10:0 was not detected in 50 (47.2%) of 106 non-fasting blood samples, while 29 (27.4%) plasma samples contained FA10:0 less than or equal to 0.5 µmol/L (0.4 ± 0.1), and 27 (25.5%) contained it at more than 0.5 µmol/L (0.9 ± 0.3). A half of the non-fasting plasma samples contained detectable FA10:0. This simple, precise, and accurate high-performance liquid chromatography method might be useful for monitoring plasma FA10:0 during medium-chain triglycerides therapy. © The Author(s) 2015.

  8. [Determination of oxaprozin in human plasma with high performance liquid chromatography (HPLC) and its application].

    Science.gov (United States)

    Mao, Mian; Wang, Ling; Jiang, Xuehua; Yang, Lin

    2013-06-01

    The present research was aimed to develop a high performance liquid chromatography (HPLC) method to determine oxaprozin in plasma and to evaluate the bioavailability of two oxaprozin enteric coated tablets. A C18 column was used to separate the plasma after protein precipitation and the mobile phase was methanol-12. 5mmol/L ammonium acetate buffer solution (pH=3.0)(71:29). The calibration curve was linear in the concentration range of 0. 50-70. 56 microg . mL-1, and the intra and inter-day RSDs were less than 12. 33% and 10. 42% respectively. A single dose of 0. 4 g reference preparation or test preparation of oxaprozin enteric coated tablets was administered to 20 healthy volunteers according to a randomized crossover study. AUC0-->264h were (4 917. 44 +/- 629. 57) microg . h . mL-1 and (4 604. 30+/-737. 83) microg . h . mL-1, respectively; Cmax were (52. 34+/-7. 68) microg . mL-1 and (48. 66+/-4. 87) microg . mL-1, respectively; Tmax were (18. 70+/-2.27) h and (19. 30+/-1. 63) h, respectively; The relative bioavailability of test preparation was 94.0% +/- 13. 7%. The method is simple, rapid and selective for oxaprozin determination. There is no significant difference in the main pharmacokinetic parameters between the test formulation and reference formulation and the two formulations are in bioequivalence.

  9. The art and science of forming packed analytical high-performance liquid chromatography columns.

    Science.gov (United States)

    Kirkland, J J; Destefano, J J

    2006-09-08

    Columns of packed particles still are the most popular devices for high-performance liquid chromatography (HPLC) separations because of their great utility, excellent performance and wide variety. However, the forming of packed beds for efficient, stable columns traditionally has been an art where the basics of how to form optimum beds generally was not well understood. The recent development of monolith rods was introduced in part to overcome the difficulty of producing stable beds of packing particles. However, these materials are less versatile than packed particle columns. Technology developments in recent years have produced a better understanding among those skilled in the practice of how to form optimized packed beds, and this has led to widely available, high-quality commercial columns. This presentation discusses the developments that led to the present state of column packing technology. Important steps in the packing of efficient, stable beds are described. The key step of selecting the best solvent for the slurry packing method is emphasized. Factors affecting the mechanical stability of packed columns also are discussed. The early art of packing columns now has evolved into a more scientific approach that allows the packing of good columns with a minimum of effort and time.

  10. Ultra-high performance size-exclusion chromatography in polar solvents.

    Science.gov (United States)

    Vancoillie, Gertjan; Vergaelen, Maarten; Hoogenboom, Richard

    2016-12-23

    Size-exclusion chromatography (SEC) is amongst the most widely used polymer characterization methods in both academic and industrial polymer research allowing the determination of molecular weight and distribution parameters, i.e. the dispersity (Ɖ), of unknown polymers. The many advantages, including accuracy, reproducibility and low sample consumption, have contributed to the worldwide success of this analytical technique. The current generation of SEC systems have a stationary phase mostly containing highly porous, styrene-divinylbenzene particles allowing for a size-based separation of various polymers in solution but limiting the flow rate and solvent compatibility. Recently, sub-2μm ethylene-bridged hybrid (BEH) packing materials have become available for SEC analysis. These packing materials can not only withstand much higher pressures up to 15000psi but also show high spatial stability towards different solvents. Combining these BEH columns with the ultra-high performance LC (UHPLC) technology opens up UHP-SEC analysis, showing strongly reduced runtimes and unprecedented solvent compatibility. In this work, this novel characterization technique was compared to conventional SEC using both highly viscous and highly polar solvents as eluent, namely N,N-dimethylacetamide (DMAc), N,N-dimethylformamide (DMF) and methanol, focusing on the suitability of the BEH-columns for analysis of highly functional polymers. The results show a high functional group compatibility comparable with conventional SEC with remarkably short runtimes and enhanced resolution in methanol. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Identification and quantitation of asparagine and citrulline using high-performance liquid chromatography (HPLC).

    Science.gov (United States)

    Bai, Cheng; Reilly, Charles C; Wood, Bruce W

    2007-03-28

    High-performance liquid chromatography (HPLC) analysis was used for identification of two problematic ureides, asparagine and citrulline. We report here a technique that takes advantage of the predictable delay in retention time of the co-asparagine/citrulline peak to enable both qualitative and quantitative analysis of asparagine and citrulline using the Platinum EPS reverse-phase C18 column (Alltech Associates). Asparagine alone is eluted earlier than citrulline alone, but when both of them are present in biological samples they may co-elute. HPLC retention times for asparagine and citrulline were influenced by other ureides in the mixture. We found that at various asparagines and citrulline ratios [= 3:1, 1:1, and 1:3; corresponding to 75:25, 50:50, and 25:75 (microMol ml(-1)/microMol ml(-1))], the resulting peak exhibited different retention times. Adjustment of ureide ratios as internal standards enables peak identification and quantification. Both chemicals were quantified in xylem sap samples of pecan [Carya illinoinensis (Wangenh.) K. Koch] trees. Analysis revealed that tree nickel nutrition status affects relative concentrations of Urea Cycle intermediates, asparagine and citrulline, present in sap. Consequently, we concluded that the HPLC methods are presented to enable qualitative and quantitative analysis of these metabolically important ureides.

  12. Improved assay for thymine base damage in E. coli using high performance liquid chromatography

    International Nuclear Information System (INIS)

    Claycamp, H.G.

    1985-01-01

    A simple high performance liquid chromatography (HPLC) technique has been established for the simultaneous assay of thymine and thymidine radiation damage products. The HPLC procedure uses an isocratic mobile phase of 4% acetonitrile in 0.2 M ammonium dihydrogen phosphate (pH 5.0), a reversed-phase octadecylsilicate (5 micro-spherical packing) 0.45 x 25 cm column, and a variable wavelength UV detector. This procedure affords much better resolution than other published procedures that use 10 micron columns or separate assays for bases and nucleosides. For example, irradiation of 5 x 10 -3 M thymidine solutions have been performed to calibrate the system for base damage assays in E. coli. This yields up to 15 resolvable residues within 20 minutes. Sensitivity of the system (at 2210 nm) for 5,6- dihydrothymine (DHT) is about 10 -10 moles. Preliminary results show that this translates to about 0.4 DHT residues per 10 6 daltons of E. coli DNA. This is comparable to the sensitivities of monoclonal assays to thymine damage products that have recently been reported by others. Since it is feasible that the sensitivity of this system can be improved by 2-3 times, this HPLC technique should provide a simple and rapid means of detecting E. coli base damage release and base damage in nucleoside hydrolysates of DNA

  13. Simultaneous Determination of Four Preservatives in Foodstuffs by High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Mohammad Faraji

    2016-04-01

    Full Text Available Background and objectives:  High concentration of preservatives in food may result in gastrointestinal disturbances whereby some patients suffering from asthma, rhinitis, or urticaria. The aim of this study is the introduction and optimization a new method for simultaneous determination of four preservatives (SB, PS, MP, PP in foodstuff by high performance liquid chromatography. Materials and methods: Important factors in extraction, separation and determination process were optimized using the one variable at a time method.  Figures of merit of the proposed method were evaluated. The amount of SB, PS, MP, PP in some food samples were determined using the proposed method. Result: The results showed that the obtained chromatogram of extract was free of significant interferences. The preservatives recoveries ranged from 88% to 110 %. Concentration of SB, PS, MP and PP in the 20studied samples ranges between N.D-639.9, N.D -214.5, N.D -579.8 and N.D -30.5 mg kg-1, respectively  Conclusion: The performance and reliability of proposed method as a simple, efficient and fast method for determination of SB, PS, MP, PP in the food samples was demonstrated.

  14. Evaluation of Some Pesticide Residues in Fruits import by High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Lamees Munef Abdul-Latef

    2017-02-01

    Full Text Available Pesticide residues have been found in various fruits and vegetables.  This studycollected 24 samples and reported a method based on High Performance Liquid Chromatography (HPLC. For determination of pesticide residues used in Some fruits which were collected from different markets of Baghdad city to make 24 samples from (peel , core, mixture for each type of fruits markets as:  Oranges( Egypt , Africa, Pomegranate ( Egypt, Mango ( Kenya , Pears (China,  Plum fruits ( Africa , Kiwi (Turkey .That detective of ( 5 different  pesticides (diazinon,  malathion, chlorpyrifos ,parathion  and cypermethrin. The results were detected of multi-residues of  pesticides on the fruit in ( peel , core , mixture may be in the limit of Maximum residue limits(MRL or higher of it   . The pesticides detected that exceeding the limits are: cypermethrin  in Kiwi (peel, core and mixtureat(0.204,0.038, 0.537, in pomegranate detected in (peel and mixture at (0.509,0.189 mg/kg , Diazinondetccted in Egyptian orange in (peel ,core and mixture at (0.031,0.207, 0.099 mg/kg . Malathion and Parathion was not detected at any type of fruits and  that results was compared with codex of FAO/WHO(2013. That data is important to monitor residues in food and to fill gaps in current knowledge would be helpful in assessing human exposure risks from ingestion of contaminated Fruits Imported to our country .

  15. High performance liquid chromatography determination of theobromine and caffeine in cocoa beans gamma irradiated

    International Nuclear Information System (INIS)

    Soares, Anderson D.B.; Mansur Neto, Elias

    1997-01-01

    Irradiation is a processing technology that has been shown to be a wholesome process by many scientific studies conducted worldwide during the past 40 years, which has been approved by 37 countries. Irradiated foods have been studied so extensively, that the effects on foods are better understood than any other preservation process, including food freezing and dehydration. Cocoa beans has been commercially irradiated in countries such as Ivory Coast and Argentina. The alkaloids theobromine and caffeine are responsible for the mildly stimulating properties and bitter taste of cocoa and chocolate products. Previously fermented dried and nonfumigated cocoa beans were irradiated at doses of 0, 5.0, 10.0 and 20.0 kGy using Co-60 gamma rays. The samples were analysed for determining theobromine and caffeine contents in the cocoa beans by TIMBIE et al. (1978) high performance liquid chromatography (HPLC) method. Boiling water extracts were cooled, centrifuged and injected into the chromatograph. Theobromine and caffeine were quantitated at 273 nm and showed the tendency of decreasing as the dose of radiation increases. Theobromine and caffeine ranged from 42.3 to 37.1 mg/g and from 7.60 to 6.13 mg/g. respectively from 0 to 20.0 kGy. These results were discussed in relation to the possible acceptance of radiosterization of cocoa beans commercially up to the dose of 20.0 kGy. (author). 10 refs., 1 tab

  16. Surface-bonded ionic liquid stationary phases in high-performance liquid chromatography--a review.

    Science.gov (United States)

    Pino, Verónica; Afonso, Ana M

    2012-02-10

    Ionic liquids (ILs) are a class of ionic, nonmolecular solvents which remain in liquid state at temperatures below 100°C. ILs possess a variety of properties including low to negligible vapor pressure, high thermal stability, miscibility with water or a variety of organic solvents, and variable viscosity. IL-modified silica as novel high-performance liquid chromatography (HPLC) stationary phases have attracted considerable attention for their differential behavior and low free-silanol activity. Indeed, around 21 surface-confined ionic liquids (SCIL) stationary phases have been developed in the last six years. Their chromatographic behavior has been studied, and, despite the presence of a positive charge on the stationary phase, they showed considerable promise for the separation of neutral solutes (not only basic analytes), when operated in reversed phase mode. This aspect points to the potential for truly multimodal stationary phases. This review attempts to summarize the state-of-the-art about SCIL phases including their preparation, chromatographic behavior, and analytical performance. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. Highly sensitive assay for tyrosine hydroxylase activity by high-performance liquid chromatography.

    Science.gov (United States)

    Nagatsu, T; Oka, K; Kato, T

    1979-07-21

    A highly sensitive assay for tyrosine hydroxylase (TH) activity by high-performance liquid chromatography (HPLC) with amperometric detection was devised based on the rapid isolation of enzymatically formed DOPA by a double-column procedure, the columns fitted together sequentially (the top column of Amberlite CG-50 and the bottom column of aluminium oxide). DOPA was adsorbed on the second aluminium oxide column, then eluted with 0.5 M hydrochloric acid, and assayed by HPLC with amperometric detection. D-Tyrosine was used for the control. alpha-Methyldopa was added to the incubation mixture as an internal standard after incubation. This assay was more sensitive than radioassays and 5 pmol of DOPA formed enzymatically could be measured in the presence of saturating concentrations of tyrosine and 6-methyltetrahydropterin. The TH activity in 2 mg of human putamen could be easily measured, and this method was found to be particularly suitable for the assay of TH activity in a small number of nuclei from animal and human brain.

  18. Determination of Flurbiprofen in Human Plasma by High-Performance Liquid Chromatography.

    Science.gov (United States)

    Yilmaz, Bilal; Erdem, Ali Fuat

    2015-10-01

    A simple high-performance liquid chromatography method has been developed for determination of flurbiprofen in human plasma. The method was validated on an Ace C18 column using UV detection. The mobile phase was acetonitrile-0.05 M potassium dihydrogen phosphate solution (60:40, v/v) adjusted to pH 3.5 with phosphoric acid. The calibration curve was linear between the concentration range of 0.10-5.0 μg/mL. Intra- and inter-day precision values for flurbiprofen in plasma were flurbiprofen from human plasma were between 93.0 and 98.9%. The limits of detection and quantification of flurbiprofen were 0.03 and 0.10 μg/mL, respectively. In addition, this assay was applied to determine the pharmacokinetic parameters of flurbiprofen in six healthy Turkish volunteers who had been given 100 mg flurbiprofen. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  19. Speciation analysis of cobalt in foods by high-performance liquid chromatography and neutron activation analysis

    International Nuclear Information System (INIS)

    Muto, Toshio; Koyama, Motoko

    1994-01-01

    A combined method by coupling high-performance liquid chromatography (HPLC, as a separation method) with neutron activation analysis (as a detection method) have been applied to the speciation analysis of cobalt in daily foods (e.g. egg, fish and milk). Cobalt species including free cobalt, vitamin B 12 and protein-bound cobalt were separated with a preparative HPLC and a centrifuge. Subsequently, the determination of cobalt in the separated species was made by neutron activation analysis. The results showed that the content of the total cobalt in the foods was found to lie in the range 0.4-11ng/g(0.4-11ppb) based on wet weight. The compositions of free cobalt, vitamin B 12 and protein-bound cobalt were ranged 16-43%, 55-73%, 2.3-17%, respectively. These experimental evidences suggest that the combination of HPLC and neutron activation analysis is expected to be a useful tool for speciation analysis of trace elements in biological as well as environmental materials. (author)

  20. Methacrylate-bonded covalent-organic framework monolithic columns for high performance liquid chromatography.

    Science.gov (United States)

    Liu, Li-Hua; Yang, Cheng-Xiong; Yan, Xiu-Ping

    2017-01-06

    Covalent-organic frameworks (COFs) are a newfangled class of intriguing microporous materials. Considering their unique properties, COFs should be promising as packing materials for high performance liquid chromatography (HPLC). However, the irregular shape and sub-micrometer size of COFs synthesized via the traditional methods render the main obstacles for the application of COFs in HPLC. Herein, we report the preparation of methacrylate-bonded COF monolithic columns for HPLC to overcome the above obstacles. The prepared COF bonded monolithic columns not only show good homogeneity and permeability, but also give high column efficiency, good resolution and precision for HPLC separation of small molecules including polycyclic aromatic hydrocarbons, phenols, anilines, nonsteroidal anti-inflammatory drugs and benzothiophenes. Compared with the bare polymer monolithic column, the COF bonded monolithic columns show enhanced hydrophobic, π-π and hydrogen bond interactions in reverse phase HPLC. The results reveal the great potential of COF bonded monoliths for HPLC and COFs in separation sciences. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Analysis of metalaxyl racemate using high performance liquid chromatography coupled with four kinds of detectors.

    Science.gov (United States)

    Chen, Tao; Fan, Jun; Gao, Ruiqi; Wang, Tai; Yu, Ying; Zhang, Weiguang

    2016-10-07

    Chiral stationary phase-high performance liquid chromatography coupled with various detectors has been one of most commonly used methods for analysis and separation of chiral compounds over the past decades. Various detectors exhibit different characteristics in qualitative and quantitative studies under different chromatographic conditions. Herein, a comparative evaluation of HPLC coupled with ultraviolet, optical rotation, refractive index, and evaporative light scattering detectors has been conducted for qualitative and quantitative analyses of metalaxyl racemate. Effects of separation conditions on the peak area ratio between two enantiomers, including sample concentration, column temperature, mobile phase composition, as well as flow rate, have been investigated in detail. In addition, the limits of detection, the limits of quantitation, quantitative range and precision for these two enantiomers by using four detectors have been also studied. As indicated, the chromatographic separation conditions have been slight effects on ultraviolet and refractive index detections and the peak area ratio between two enantiomers remains almost unchanged, but the evaporative light scattering detection has been significantly affected by the above-mentioned chromatographic conditions and the corresponding peak area ratios varied greatly. Moreover, the limits of detection, the limits of quantitation, and the quantitative ranges of two enantiomers with UV detection were remarkably lower by 1-2 magnitudes than the others. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Quality Evaluation of Potentilla fruticosa L. by High Performance Liquid Chromatography Fingerprinting Associated with Chemometric Methods.

    Science.gov (United States)

    Liu, Wei; Wang, Dongmei; Liu, Jianjun; Li, Dengwu; Yin, Dongxue

    2016-01-01

    The present study was performed to assess the quality of Potentilla fruticosa L. sampled from distinct regions of China using high performance liquid chromatography (HPLC) fingerprinting coupled with a suite of chemometric methods. For this quantitative analysis, the main active phytochemical compositions and the antioxidant activity in P. fruticosa were also investigated. Considering the high percentages and antioxidant activities of phytochemicals, P. fruticosa samples from Kangding, Sichuan were selected as the most valuable raw materials. Similarity analysis (SA) of HPLC fingerprints, hierarchical cluster analysis (HCA), principle component analysis (PCA), and discriminant analysis (DA) were further employed to provide accurate classification and quality estimates of P. fruticosa. Two principal components (PCs) were collected by PCA. PC1 separated samples from Kangding, Sichuan, capturing 57.64% of the variance, whereas PC2 contributed to further separation, capturing 18.97% of the variance. Two kinds of discriminant functions with a 100% discrimination ratio were constructed. The results strongly supported the conclusion that the eight samples from different regions were clustered into three major groups, corresponding with their morphological classification, for which HPLC analysis confirmed the considerable variation in phytochemical compositions and that P. fruticosa samples from Kangding, Sichuan were of high quality. The results of SA, HCA, PCA, and DA were in agreement and performed well for the quality assessment of P. fruticosa. Consequently, HPLC fingerprinting coupled with chemometric techniques provides a highly flexible and reliable method for the quality evaluation of traditional Chinese medicines.

  3. [Determination of lutein in infant formula milk powder using ultra-high performance liquid chromatography].

    Science.gov (United States)

    Wang, Lin; Huang, Junrong; Zhang, Li; Feng, Feng; Ling, Yun; Chu, Xiaogang; Li, Hongliang

    2013-12-01

    An ultra-high performance liquid chromatography (U-HPLC) method for the determination of lutein in the infant formula milk powder was developed. The sample was extracted with acetone and defatted using freezing centrifugation method. The U-HPLC separation was achieved using a YMC Carotenoid C30 column (150 mm x 4.6 mm, 3 microm) with the mixture of methanol/methyl tert-butyl ether (70: 30, v/v) as the mobile phase under isocratic elution. The flow rate was 0.5 mL/min and the column oven temperature was 25 degrees C. The injection volume was 5 microL. It was detected on a photodiode array detector at a wavelength of 445 nm. The results showed that the linear range was 20-500 microg/L (r = 0.9999), and the limit of quantification was 20 microg/L. The mean recoveries of lutein varied from 97.9% to 104.4% spiked at 50, 250 and 2,000 microg/kg. The established method is simple, accurate and sensitive for the rapid determination of lutein in infant formula milk powder.

  4. Quantitative structure-retention relationships of pesticides in reversed-phase high-performance liquid chromatography

    International Nuclear Information System (INIS)

    Aschi, Massimiliano; D'Archivio, Angelo Antonio; Maggi, Maria Anna; Mazzeo, Pietro; Ruggieri, Fabrizio

    2007-01-01

    In this paper, a quantitative structure-retention relationships (QSRR) method is employed to predict the retention behaviour of pesticides in reversed-phase high-performance liquid chromatography (HPLC). A six-parameter nonlinear model is developed by means of a feed-forward artificial neural network (ANN) with back-propagation learning rule. Accurate description of the retention factors of 26 compounds including commonly used insecticides, herbicides and fungicides and some metabolites is successfully achieved. In addition to the acetonitrile content, included to describe composition of the water-acetonitrile mobile phase, the octanol-water partition coefficient (from literature) and four quantum chemical descriptors are considered to account for the effect of solute structure on the retention. These are: the total dipole moment, the mean polarizability, the anisotropy of polarizability and a descriptor of hydrogen bonding ability based on the atomic charges on hydrogen bond donor and acceptor chemical functionalities. The proposed nonlinear QSRR model exhibits a high degree of correlation between observed and computed retention factors and a good predictive performance in wide range of mobile phase composition (40-65%, v/v acetonitrile) that supports its application for the prediction of the chromatographic behaviour of unknown pesticides. A multilinear regression model based on the same six descriptors shows a significantly worse predictive capability

  5. Quantitative structure-retention relationships of pesticides in reversed-phase high-performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Aschi, Massimiliano [Dipartimento di Chimica, Ingegneria Chimica e Materiali, Universita degli Studi di L' Aquila, Via Vetoio, 67010 Coppito, L' Aquila (Italy); D' Archivio, Angelo Antonio [Dipartimento di Chimica, Ingegneria Chimica e Materiali, Universita degli Studi di L' Aquila, Via Vetoio, 67010 Coppito, L' Aquila (Italy)]. E-mail: darchivi@univaq.it; Maggi, Maria Anna [Dipartimento di Chimica, Ingegneria Chimica e Materiali, Universita degli Studi di L' Aquila, Via Vetoio, 67010 Coppito, L' Aquila (Italy); Mazzeo, Pietro [Dipartimento di Chimica, Ingegneria Chimica e Materiali, Universita degli Studi di L' Aquila, Via Vetoio, 67010 Coppito, L' Aquila (Italy); Ruggieri, Fabrizio [Dipartimento di Chimica, Ingegneria Chimica e Materiali, Universita degli Studi di L' Aquila, Via Vetoio, 67010 Coppito, L' Aquila (Italy)

    2007-01-23

    In this paper, a quantitative structure-retention relationships (QSRR) method is employed to predict the retention behaviour of pesticides in reversed-phase high-performance liquid chromatography (HPLC). A six-parameter nonlinear model is developed by means of a feed-forward artificial neural network (ANN) with back-propagation learning rule. Accurate description of the retention factors of 26 compounds including commonly used insecticides, herbicides and fungicides and some metabolites is successfully achieved. In addition to the acetonitrile content, included to describe composition of the water-acetonitrile mobile phase, the octanol-water partition coefficient (from literature) and four quantum chemical descriptors are considered to account for the effect of solute structure on the retention. These are: the total dipole moment, the mean polarizability, the anisotropy of polarizability and a descriptor of hydrogen bonding ability based on the atomic charges on hydrogen bond donor and acceptor chemical functionalities. The proposed nonlinear QSRR model exhibits a high degree of correlation between observed and computed retention factors and a good predictive performance in wide range of mobile phase composition (40-65%, v/v acetonitrile) that supports its application for the prediction of the chromatographic behaviour of unknown pesticides. A multilinear regression model based on the same six descriptors shows a significantly worse predictive capability.

  6. [Determination of emamectin benzoate residue in vegetables by high performance liquid chromatography with fluorescence detection].

    Science.gov (United States)

    Zhang, Yan; Wu, Yinliang; Hu, Jiye; Wang, Hongwei; Pan, Canping; Liu, Fengmao

    2008-01-01

    A method was developed for the determination of emamectin benzoate residue in cabbage and mushroom using solid-phase extraction (SPE) and high performance liquid chromatography (HPLC) with fluorescence detection. The sample was extracted with ethyl acetate. Further cleanup was performed on a propylsulfonic acid solid phase extraction cartridge, followed by the derivatization with trifluoroacetic anhydride in the presence of N-methylimidazole. The amount of derivatized emamectin benzoate was determined by fluorescence detector after separation by HPLC. The detection limit was 0.10 microg/kg for cabbage and mushroom samples. The recoveries of emamectin benzoate in cabbage and mushroom samples were 78.6%-84.9%. The inter-day relative standard deviation (RSD) and intra-day RSD were 2.7%-6.0% and 3.1%-8.9%, respectively, at the fortified levels of 1.0-20.0 microg/kg. The calibration curve of emamectin benzoate in vegetables at the concentration range of 0.002 mg/L to 0.10 mg/L was linear (r = 0.9999).

  7. Analysis of serotonin concentrations in human milk by high-performance liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Chiba, Takeshi; Maeda, Tomoji; Tairabune, Tomohiko; Tomita, Takashi; Sanbe, Atsushi; Takeda, Rika; Kikuchi, Akihiko; Kudo, Kenzo

    2017-03-25

    Serotonin (5-hydroxytryptamine, 5-HT) plays an important role in milk volume homeostasis in the mammary gland during lactation; 5-HT in milk may also affect infant development. However, there are few reports on 5-HT concentrations in human breast milk. To address this issue, we developed a simple method based on high-performance liquid chromatography with fluorescence detection (HPLC-FD) for measuring 5-HT concentrations in human breast milk. Breast milk samples were provided by four healthy Japanese women. Calibration curves for 5-HT in each sample were prepared with the standard addition method between 5 and 1000 ng/ml, and all had correlation coefficients >0.999. The recovery of 5-HT was 96.1%-101.0%, with a coefficient of variation of 3.39%-8.62%. The range of 5-HT concentrations estimated from the calibration curves was 11.1-51.1 ng/ml. Thus, the HPLC-FD method described here can effectively extract 5-HT from human breast milk with high reproducibility. Copyright © 2017 Elsevier Inc. All rights reserved.

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

    International Nuclear Information System (INIS)

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

    2013-01-01

    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 r 2 = 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).

  9. Quality Evaluation of Potentilla fruticosa L. by High Performance Liquid Chromatography Fingerprinting Associated with Chemometric Methods

    Science.gov (United States)

    Liu, Wei; Wang, Dongmei; Liu, Jianjun; Li, Dengwu; Yin, Dongxue

    2016-01-01

    The present study was performed to assess the quality of Potentilla fruticosa L. sampled from distinct regions of China using high performance liquid chromatography (HPLC) fingerprinting coupled with a suite of chemometric methods. For this quantitative analysis, the main active phytochemical compositions and the antioxidant activity in P. fruticosa were also investigated. Considering the high percentages and antioxidant activities of phytochemicals, P. fruticosa samples from Kangding, Sichuan were selected as the most valuable raw materials. Similarity analysis (SA) of HPLC fingerprints, hierarchical cluster analysis (HCA), principle component analysis (PCA), and discriminant analysis (DA) were further employed to provide accurate classification and quality estimates of P. fruticosa. Two principal components (PCs) were collected by PCA. PC1 separated samples from Kangding, Sichuan, capturing 57.64% of the variance, whereas PC2 contributed to further separation, capturing 18.97% of the variance. Two kinds of discriminant functions with a 100% discrimination ratio were constructed. The results strongly supported the conclusion that the eight samples from different regions were clustered into three major groups, corresponding with their morphological classification, for which HPLC analysis confirmed the considerable variation in phytochemical compositions and that P. fruticosa samples from Kangding, Sichuan were of high quality. The results of SA, HCA, PCA, and DA were in agreement and performed well for the quality assessment of P. fruticosa. Consequently, HPLC fingerprinting coupled with chemometric techniques provides a highly flexible and reliable method for the quality evaluation of traditional Chinese medicines. PMID:26890416

  10. High-performance liquid chromatography of quinoidal imminium compounds derived from triphenylmethanes

    Science.gov (United States)

    Abidi, S.L.

    1983-01-01

    A series of eleven p-aminotriphenylmethane dyes have been studied by high-performance liquid chromatography (HPLC). The combined use of HPLC and spectrophotometry permits specific detection of these compounds in the visible range around 600 nm. As the high affinity of the imminium cations for the active sites of the hydrocarbonaceous stationary phase has presented difficulties for reversed-phase HPLC with pure solvents, organic electrolytes were added to the mobile phase to facilitate the elution of the components with improved selectivity, sensitivity (minimum detection limit, 0.1 μg/ml), and peak symmetry. The effects of chromatographic variables on the component retentivity were investigated. Retention times of the dye analytes decreased with increasing concentration of the added ionic reagent and with decreasing number of the hydrophobic alkyl substituents on the nitrogen atom. The influence of pH on the retention parameters appears to parallel that observed previously for cationic quaternary ammonium compounds. Among the acidic reagents employed, naphthalenesulfonic acid yielded the most satisfactory results. The use of binary electrolyte systems invariably improved the chromatographic behavior of the imminium solutes analyzed. Results obtained with two different octadecylsilica columns have been compared.

  11. Applications of plasma spectrometry and high performance liquid chromatography in environmental and food science

    International Nuclear Information System (INIS)

    Iordache, Andreea-Maria; Biraruti, Elisabeta-Irina; Ionete, Roxana-Elena

    2008-01-01

    Full text: Plasma spectrometry has many applications in food science in analysis of a wide range of samples in the food chain. Food science in the broadest sense can be extended to include soil chemistry, plant uptake and, at the other end of the food chain, studies into the metabolic fate of particular elements or elemental species when the foods are consumed by humans or animals. Inductively Coupled Plasma Mass Spectrometry allows multi-element measurements of most elements in the periodic table. A very sensitive analytical technique for trace analysis of samples can be performed by inductively plasma mass spectrometer with quadrupolar detector using ultrasonic nebulization. High Performance Liquid Chromatography (HPLC) is an analytical technique for the separation and determination of organic and inorganic solutes in any samples especially biological, pharmaceutical, food, environmental. The present paper emphasizes that the future tendencies HPLC-ICP-MS is often the preferred analytical technique for these applications due to the simplicity of the coupling between the HPLC and ICP-MS Varian 820 using ultrasonic nebulization, potential for on-line separations with high species specificity and the capability for optimum limits of detection without the necessity of using complex hydride generation mechanisms. (authors)

  12. Determination of Diclofenac sodium by high-performance liquid chromatography in 0,1 % eyedrops

    International Nuclear Information System (INIS)

    Garcia Penna, Caridad Margarita; Pereda Rodriguez, Diana; Gonzalez Hortezon, Ania; Montes de Oca Porto, Yanet; Cannizares Arencibia, Yanara; Leon Guerrero, Gissel Maria

    2009-01-01

    Diclofenac sodium is a drug prescribed in eye anterior segment inflammation postoperative treatments, preoperative and postoperative cataract meiosis inhibition, symptomatic treatment of non-infectious chronic conjunctivitis, of ocular inflammation, of ocular pain, and of refractory postsurgical photophobia. In present paper we developed and validated an analytical method by high-performance liquid chromatography to quality control, and stability studies of Diclofenac sodium of 1 % eye drops. Method was based on active principle separation through endcapped 1 00RP-8 Lichrospher chromatographic column (5 μ) (250 x 4 mm) using UV detection to 254 nm, thus using a mobile phase including sodium a solution of hydrogen phosphate to a 2.5 pH: methanol in a 34:60 ratio. Calibration curve was plotted during the interval of 60 to 140 % where it was linear with a correlation coefficient equal to 0.9995; statistic test for interception and slope was considered as non-significant. We achieved a recovery rate of 100.25 % during study concentration interval, and Cochran (G) and Student (t) tests were not significant. Variation coefficient in repeating study was equal to 0.39 % for the 6 assayed replica, whereas in average precision analysis Fischer and Student tests were non-significant. Analytical method was linear, precise, specific and exact during the interval of study concentrations. (Author)

  13. Blind column selection protocol for two-dimensional high performance liquid chromatography.

    Science.gov (United States)

    Burns, Niki K; Andrighetto, Luke M; Conlan, Xavier A; Purcell, Stuart D; Barnett, Neil W; Denning, Jacquie; Francis, Paul S; Stevenson, Paul G

    2016-07-01

    The selection of two orthogonal columns for two-dimensional high performance liquid chromatography (LC×LC) separation of natural product extracts can be a labour intensive and time consuming process and in many cases is an entirely trial-and-error approach. This paper introduces a blind optimisation method for column selection of a black box of constituent components. A data processing pipeline, created in the open source application OpenMS®, was developed to map the components within the mixture of equal mass across a library of HPLC columns; LC×LC separation space utilisation was compared by measuring the fractional surface coverage, fcoverage. It was found that for a test mixture from an opium poppy (Papaver somniferum) extract, the combination of diphenyl and C18 stationary phases provided a predicted fcoverage of 0.48 and was matched with an actual usage of 0.43. OpenMS®, in conjunction with algorithms designed in house, have allowed for a significantly quicker selection of two orthogonal columns, which have been optimised for a LC×LC separation of crude extractions of plant material. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Determination of aristolochic acids by high-performance liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Wang, Yinan; Chan, Wan

    2014-06-25

    Nephrotoxic and carcinogenic aristolochic acids (AAs) are naturally occurring nitrophenanthrene carboxylic acids in the herbal genus Aristolochia. The misuse of AA-containing herbs in preparing slimming drugs has caused hundred of cases of kidney disease in Belgium women in a slimming regime in the early 1990s. Accumulating evidence also suggested that prolong dietary intake of AA-contaminated food is one of the major causes to the Balkan endemic nephropathy that was first observed in the late 1950s. Therefore, analytical methods of high sensitivity are extremely important for safeguarding human exposure to AA-containing herbal medicines, herbal remedies, and food composites. In this paper, we describe the development of a new high-performance liquid chromatography coupled fluorescence detector (HPLC-FLD) method for the sensitive determination of AAs. The method makes use of a novel cysteine-induced denitration reaction that "turns on" the fluorescence of AAs for fluorometric detections. Our results showed that the combination of cysteine-induced denitration and HPLC-FLD analysis allows for sensitive quantification of AA-I and AA-II at detection limits of 27.1 and 25.4 ng/g, respectively. The method was validated and has been successfully applied in quantifying AAs in Chinese herbal medicines.

  15. Quantitative Determination of Compounds from Akebia quinata by High-Performance Liquid Chromatography

    International Nuclear Information System (INIS)

    Yen, Nguyen; Thu, Nguyen; Zhao, Bing Tian; Woo, Mi Hee; Min, Byung Sun; Lee, Jae Hyun; Kim, Jeong Ah; Son, Jong Keun; Choi, Jae Sui; Woo, Eun Rhan

    2014-01-01

    To provide the scientific corroboration of the traditional uses of Akebia quinata (Thunb.) Decne., a detailed analytical examination of A. quinata stems was carried out using a reversed-phase high performance liquid chromatography (RP-HPLC) method coupled to photodiode array detector (PDA) for the simultaneous determination of four phenolic substances; cuneataside D, 2-(3,4-dihydroxyphenyl)ethyl-O-β-D-glucopyranoside, 3-caffeoylquinic acid and calceolarioside B. Particular attention was focused on the main compound, 3-caffeoylquinic acid, which has a range of biological functions. In addition, 2-(3,4-dihydroxyphenyl)ethyl-O-β-D-glucopyranoside was considered as a discernible marker of A. quinata from its easy confuse plants. The contents of compounds 2 and 3 ranged from 0.72 to 2.68 mg/g and from 1.66 to 5.64 mg/g, respectively. The validation data indicated that this HPLC/PDA assay was used successfully to quantify the four phenolic compounds in A. quinata from different locations using relatively simple conditions and procedures. The pattern-recognition analysis data from 53 samples classified them into two groups, allowing discrimination between A. quinata and comparable herbs. The results suggest that the established HPLC/PDA method is suitable for quantitation and pattern-recognition analyses for a quality evaluation of this medicinal herb

  16. Arsenic speciation in soil using high performance liquid chromatography/inductively coupled plasma/mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Bass, D.A.; Yaeger, J.S.; Parish, K.J.; Crain, J.S.; Kiely, J.T.; Gowdy, M.J. [Argonne National Lab., IL (United States); Mohrman, G.B.; Besmer, M.G. [Rocky Mountain Arsenal, Commerce City, CO (United States)

    1996-08-01

    A method has been developed to identify and quantify As(III), As(V), and organoarsenic compounds in soil samples from the Rocky Mountain Arsenal (RMA) by high performance liquid chromatography/inductively coupled plasma/mass spectrometry (HPLC/ICP/MS). The soils were extracted using tetrabutylammonium hydroxide (TBAH) and sonication. The percentages of As(III), As(V), and organoarsenic species extracted from soil samples were 30, 50, and 100 respectively. The arsenic species were not altered during the extraction process. They were separated by reversed-phase, ion-pairing, HPLC using a microbore Inertsil-ODS{trademark} column. The HPLC column effluent was introduced into an ICP/MS system using a direct injection nebulizer (DIN). Detection limits of less than 1 pg were readily obtained for each arsenic species. Internal standards are recommended to increase accuracy and precision. Soil samples spiked with arsenic oxide, sodium arsenate, dimethylarsinic acid (DMAA), and chlorovinyl arsenious acid (CVAA) were extracted, identified and quantified with the HPLC/ICP/MS system. The soil samples were analyzed in support of the analytical needs of a thermal desorption treatability study being conducted at the RMA.

  17. Quantitative Determination of Compounds from Akebia quinata by High-Performance Liquid Chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Yen, Nguyen; Thu, Nguyen; Zhao, Bing Tian; Woo, Mi Hee; Min, Byung Sun [Catholic Univ. of Daegu, Gyeongsan (Korea, Republic of); Lee, Jae Hyun [Dongguk Univ., Yongin (Korea, Republic of); Kim, Jeong Ah [Kyungpook National Univ., Daegu (Korea, Republic of); Son, Jong Keun [Yeungnam Univ., Gyeongsan (Korea, Republic of); Choi, Jae Sui [Pukyung National Univ., Busan (Korea, Republic of); Woo, Eun Rhan [Chosun Univ., Gwangju (Korea, Republic of)

    2014-07-15

    To provide the scientific corroboration of the traditional uses of Akebia quinata (Thunb.) Decne., a detailed analytical examination of A. quinata stems was carried out using a reversed-phase high performance liquid chromatography (RP-HPLC) method coupled to photodiode array detector (PDA) for the simultaneous determination of four phenolic substances; cuneataside D, 2-(3,4-dihydroxyphenyl)ethyl-O-β-D-glucopyranoside, 3-caffeoylquinic acid and calceolarioside B. Particular attention was focused on the main compound, 3-caffeoylquinic acid, which has a range of biological functions. In addition, 2-(3,4-dihydroxyphenyl)ethyl-O-β-D-glucopyranoside was considered as a discernible marker of A. quinata from its easy confuse plants. The contents of compounds 2 and 3 ranged from 0.72 to 2.68 mg/g and from 1.66 to 5.64 mg/g, respectively. The validation data indicated that this HPLC/PDA assay was used successfully to quantify the four phenolic compounds in A. quinata from different locations using relatively simple conditions and procedures. The pattern-recognition analysis data from 53 samples classified them into two groups, allowing discrimination between A. quinata and comparable herbs. The results suggest that the established HPLC/PDA method is suitable for quantitation and pattern-recognition analyses for a quality evaluation of this medicinal herb.

  18. Identification and Quantitation of Asparagine and Citrulline Using High-Performance Liquid Chromatography (HPLC

    Directory of Open Access Journals (Sweden)

    Cheng Bai

    2007-01-01

    Full Text Available High-performance liquid chromatography (HPLC analysis was used for identification of two problematic ureides, asparagine and citrulline. We report here a technique that takes advantage of the predictable delay in retention time of the co-asparagine/citrulline peak to enable both qualitative and quantitative analysis of asparagine and citrulline using the Platinum EPS reverse-phase C18 column (Alltech Associates. Asparagine alone is eluted earlier than citrulline alone, but when both of them are present in biological samples they may co-elute. HPLC retention times for asparagine and citrulline were influenced by other ureides in the mixture. We found that at various asparagines and citrulline ratios [= 3:1, 1:1, and 1:3; corresponding to 75:25, 50:50, and 25:75 (μMol ml–1/μMol ml–1], the resulting peak exhibited different retention times. Adjustment of ureide ratios as internal standards enables peak identification and quantification. Both chemicals were quantified in xylem sap samples of pecan [Carya illinoinensis (Wangenh. K. Koch] trees. Analysis revealed that tree nickel nutrition status affects relative concentrations of Urea Cycle intermediates, asparagine and citrulline, present in sap. Consequently, we concluded that the HPLC methods are presented to enable qualitative and quantitative analysis of these metabolically important ureides.

  19. Characterisation of brewpub beer carbohydrates using high performance anion exchange chromatography coupled with pulsed amperometric detection.

    Science.gov (United States)

    Arfelli, Giuseppe; Sartini, Elisa

    2014-01-01

    High performance anion exchange chromatography (HPAEC) coupled with pulsed amperometric detection (PAD) was optimised in order to quantify mannose, maltose, maltotriose, maltotetraose, maltopentaose, maltohexaose and maltoheptaose content of beer. The method allows the determination of above mentioned oligosaccharides, in a single chromatographic run, without any pre-treatment. Limit of detection and limit of quantification were suitable for beer. Accuracy and repeatability were good for the entire amount considered. Once optimised HPAEC PAD for the specific matrix, the second goal of this research was to verify the possibility to discriminate beers, depending on their style. The carbohydrates content of brewpub commercial beers was very variable, ranging from 19.3 to 1469mg/L (mannose), 34.5 to 2882mg/L (maltose), 141.9 to 20731mg/L (maltotriose), 168.5 to 7650mg/L (maltotetraose), 20.1 to 2537mg/L (maltopentaose), 22.9 to 3295mg/L (maltohexaose), 8.5 to 2492mg/L (maltoeptaose), even in the same style of beer. However, the carbohydrates content was useful, jointed with other compounds amount, to discriminate different styles of beer. As a matter of fact, principal component analysis put in evidence beer differences considering some fermentation conditions and colour. Copyright © 2013 Elsevier Ltd. All rights reserved.

  20. Simultaneous determination of quinolones for veterinary use by high-performance liquid chromatography with electrochemical detection.

    Science.gov (United States)

    Rodríguez Cáceres, M I; Guiberteau Cabanillas, A; Galeano Díaz, T; Martínez Cañas, M A

    2010-02-01

    A selective method based on high-performance liquid chromatography with electrochemical detection (HPLC-ECD) has been developed to enable simultaneous determination of three fluoroquinolones (FQs), namely danofloxacin (DANO), difloxacin (DIFLO) and sarafloxacin (SARA). The fluoroquinolones are separated on a Novapack C-18 column and detected in a high sensitivity amperometric cell at a potential of +0.8 V. Solid-phase extraction was used for the extraction of the analytes in real samples. The range of concentration examined varied from 10 to 150 ng g(-1) for danofloxacin, from 25 to 100 ng g(-1) for sarafloxacin and from 50 to 315 ng g(-1) for difloxacin, respectively. The method presents detection limits under 10 ng g(-1) and recoveries around 90% for the three analytes have been obtained in the experiments with fortified samples. This HPLC-ECD approach can be useful in the routine analysis of antibacterial residues being less expensive and less complicated than other more powerful tools as hyphenated techniques. 2009 Elsevier B.V. All rights reserved.

  1. Investigation of prostaglandin levels in human milk after high performance liquid chromatography purification

    International Nuclear Information System (INIS)

    Wu-Wang, C.Y.; Neu, J.

    1986-01-01

    This study was conducted to investigate five prostaglandins (PGs), i.e. PGE 2 , PGF/sub 2α/, 13-14-dihydro-15-keto-PGF/sub 2α/ (DHKF/sub 2α/), thromboxane B 2 (TXB 2 ) and 6-keto-PGF/sub 1α/), measured by (RIA) after C 18 Sep-Pak extraction and reverse phase high performance liquid chromatography (HPLC). Two trials were performed. In each trial, 3-5 mature human milk samples were pooled, acidified and extracted for PGs. The separation of PGs by HPLC was achieved by using an isocratic solvent system of acetonitrile/water (pH 3.0) (32/68, V/V). The PG levels from the two trials were determined and averaged after monitoring the recoveries. The results indicate that PGE 2 and DHKF/sub 2α/ are the two major PGs found in extracted human milk. However, after HPLC purification, no predominant PG is found and the levels of all the five PGs are much lower compared to the extracted sample. Since the immunoreactive material was also detected in HPLC fractions not within the PG peak, low levels of PG found in human milk after HPLC is likely due to the purification step removing the bulk of nonspecific immunoreactive substances present in the sample

  2. Simultaneous characterization and quantification of 17 main compounds in Rabdosia rubescens by high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Sen Guo

    2017-04-01

    Full Text Available Rabdosia rubescens is a healthy herbal tea and well-known Chinese medicinal herb. To evaluate the quality of R. rubescens from China, a high performance liquid chromatography method with dual-wavelength detection was developed and validated. The method was successfully applied for the simultaneous characterization and quantification of 17 main constituents from four different cultivation regions in China. Under optimal conditions, analysis was performed on a Luna C-18 column and gradient elution with a solvent system of acetonitrile and 0.5% (v/v acetic acid–water at a flow rate of 1.0 mL/min and wavelength of 220 nm and 280 nm. All standard calibration curves exhibited good linearity (r2 > 0.9992 within the test ranges. The precision was evaluated by intraday and interday tests, which revealed relative standard deviation values within the ranges of 0.57–2.35% and 0.52–3.40%, respectively. The recoveries were in the range of 96.37–101.66%. The relative standard deviation values for stability and repeatability were < 5%. The contents of some compounds were low and varied with different cultivars. The proposed method could serve as a prerequisite for quality control of R. rubescens materials and products.

  3. High-performance liquid chromatography method validation for determination of tetracycline residues in poultry meat

    Directory of Open Access Journals (Sweden)

    Vikas Gupta

    2014-01-01

    Full Text Available Background: In this study, a method for determination of tetracycline (TC residues in poultry with the help of high-performance liquid chromatography technique was validated. Materials and Methods: The principle step involved in ultrasonic-assisted extraction of TCs from poultry samples by 2 ml of 20% trichloroacetic acid and phosphate buffer (pH 4, which gave a clearer supernatant and high recovery, followed by centrifugation and purification by using 0.22 μm filter paper. Results: Validity study of the method revealed that all obtained calibration curves showed good linearity (r2 > 0.999 over the range of 40-4500 ng. Sensitivity was found to be 1.54 and 1.80 ng for oxytetracycline (OTC and TC. Accuracy was in the range of 87.94-96.20% and 72.40-79.84% for meat. Precision was lower than 10% in all cases indicating that the method can be used as a validated method. Limit of detection was found to be 4.8 and 5.10 ng for OTC and TC, respectively. The corresponding values of limit of quantitation were 11 and 12 ng. Conclusion: The method reliably identifies and quantifies the selected TC and OTC in the reconstituted poultry meat in the low and sub-nanogram range and can be applied in any laboratory.

  4. Simultaneous detection of water-soluble vitamins using the High Performance Liquid Chromatography (HPLC - a review

    Directory of Open Access Journals (Sweden)

    Rosemond Godbless Dadzie

    2014-01-01

    Full Text Available The water-soluble vitamins (WSV: ascorbic acid (vitamin C, thiamine (B1, riboflavin (B2, niacin (B3, panthothenic acid (B5, pyridoxine, and pyridoxal (B6, folic acid (B9, biotin(B8 , and B12 are very essential in the diet of humankind. As a result of ever increasing pressures from both consumers and legal enforcers, to specify accurately nutritive compositions of WSV that are present in food materials, many researchers have attempted to fill this niche through the provision of highly sensitive and rapid high performance liquid chromatography (HPLC procedures. In view of the health benefits of WSV, a replete of HPLC methods have been developed for simultaneous determination of their contents in nature and fortified food samples, nutritional supplements, as well as blood plasmas. The rate of losses of these vitamins during food processing and analysis, in addition to their transient dynamics, presents complexities in developing a highly sensitive HPLC procedure for their simultaneous separations and assays. This review critically assesses the different HPLC procedures developed by researchers and available in the open literature for simultaneous determination of water-soluble vitamins (WSV in dried tropical fruits materials. The study revealed that not a single chromatographic run developed by researchers can simultaneously elute all the WSV at a time. However, the HPLC procedures that are capable of determining all the WSV were coupled with electrospray ionization mass spectroscopy (ESI-MS, thus making the set-up expensive.

  5. [Determination of amitrole in agricultural products by high performance liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Li, Li; Fu, Jian; Gao, Hongliang; Ren, Haitao; Lou, Xishan; Guan, Lihui

    2010-03-01

    A high performance liquid chromatography-tandem mass spectrometry method (HPLC-MS/MS) was developed for the analysis of amitrole residues in agricultural products. The samples were extracted by 25% acetone for wheat, fish, pork and liver samples, 1% acetic acid-25% acetone for maize and peanut samples, 1% acetic acid solution for honeysuckle, the powder of ginger, the powder of bunge prickly ash and tea leaves samples, 1% acetic acid solution-dichloromethane for apple, pineapple, spinach, carrot, perilla leaves samples, respectively, followed by liquid-liquid extraction with dichloromethane. The samples were then cleaned up by PCX or Envi-Carb solid-phase extraction cartridge. The amitrole was determined and confirmed by HPLC-MS/MS. The results showed a linear relationship in the range of 0.005 -0.1 mg/kg for amitrole. The correlation coefficient was 0.999 7. The average recoveries of amitrole in wheat, maize, peanut, pineapple, apple, carrot, spinach, pork, the powder of ginger, the powder of bunge prickly ash, perilla, liver, fish, honeysuckle and tea were 67.5% - 98.1%. The relative standard deviations (RSDs) were 1.0% - 9.8%. The limits of quantitation were 10 microg/kg for wheat, maize, peanut, pineapple, apple, carrot, spinach, pork, perilla, liver, fish, honeysuckle and 20 microg/kg for the powder of bunge prickly ash, the powder of ginger and tea, respectively. The method is simple, sensitive and accurate.

  6. [Determination of sulpride in human plasma by high performance liquid chromatography].

    Science.gov (United States)

    Yu, X; Luo, Z; Tang, J; Yu, P

    1997-11-01

    This paper describes a reliable method for the pharmacokinetic study of Sulpride in human plasma by reversed-phase high performance liquid chromatography. To compensate the loss of Sulpride during the extraction procedure we used an internal standard very similar in chemical structure and UV absorbance to those of Sulpride. The mobile phase was methanol-water-acetic acid (60:30:1) with a flow rate of 1.2 mL/min. A UV detector was used at 290 nm. The linear range was 5-100 mg/L and the detectable limit was 1.0 mg/L. The recovery and RSD were 97.95%-99.96% and 2.6%-5.1% respectively. The results showed that this method is a sensitive and accurate one which makes the pharmacokinetic study of Sulpride possible. If the concentration was too low to be detected by UV monitor, a fluorescence detector could be used with the excitation wavelength at 299 nm and emission at 342 nm. We analyzed the plasma samples from 30 day-treated psychotic patients and got the satisfactory results.

  7. Determination of catechin in lotus rhizomes by high-performance liquid chromatography.

    Science.gov (United States)

    Yan, Shou-Lei; Wang, Qing-Zhang; Peng, Guang-Hua

    2009-08-01

    A novel method was developed to analyze lotus rhizome polyphenolic catechin using high-performance liquid chromatography (HPLC). The retain time of catechin was 14.72 min under the optimized condition. Mass spectrometry was further employed to qualify and quantify the purity of the catechin peak. Good linearity (R=0.9997) was obtained within the range of 50-1,000 ng. The coefficient of variance was determined as 5.2%, with a recovery rate of 97%. The detection and quantification limitations of catechin were 23 ng and 50 ng, respectively. The catechin level was 0.0025% in the lotus rhizome, and 0.011% in the knot of the lotus rhizome (Nelumbo nucifera cv. 'damao jie'). The optimized conditions of HPLC for catechin detection in the lotus rhizome matrix were as follows: the SuperlcosIL™ LC-18 analytical column (150 mm×4.6 mm, 5 µm), methanol-water-acetic acid (10:90:1, volume ratio) as the mobile phase, an UV detector at 280 nm, a flow rate of 0.8 ml/min, column temperature at 30°C, and an injection volume of 10 µl.

  8. High performance liquid chromatography determination of theobromine and caffeine in cocoa beans gamma irradiated

    Energy Technology Data Exchange (ETDEWEB)

    Soares, Anderson D.B. [Faculdade de Engenharia de Varginha, MG (Brazil); Mansur Neto, Elias [Minas Gerais Univ., Belo Horizonte, MG (Brazil)

    1997-12-01

    Irradiation is a processing technology that has been shown to be a wholesome process by many scientific studies conducted worldwide during the past 40 years, which has been approved by 37 countries. Irradiated foods have been studied so extensively, that the effects on foods are better understood than any other preservation process, including food freezing and dehydration. Cocoa beans has been commercially irradiated in countries such as Ivory Coast and Argentina. The alkaloids theobromine and caffeine are responsible for the mildly stimulating properties and bitter taste of cocoa and chocolate products. Previously fermented dried and nonfumigated cocoa beans were irradiated at doses of 0, 5.0, 10.0 and 20.0 kGy using Co-60 gamma rays. The samples were analysed for determining theobromine and caffeine contents in the cocoa beans by TIMBIE et al. (1978) high performance liquid chromatography (HPLC) method. Boiling water extracts were cooled, centrifuged and injected into the chromatograph. Theobromine and caffeine were quantitated at 273 nm and showed the tendency of decreasing as the dose of radiation increases. Theobromine and caffeine ranged from 42.3 to 37.1 mg/g and from 7.60 to 6.13 mg/g. respectively from 0 to 20.0 kGy. These results were discussed in relation to the possible acceptance of radiosterization of cocoa beans commercially up to the dose of 20.0 kGy. (author). 10 refs., 1 tab.

  9. Quantitative analysis of phylloquinone (vitamin K1) in soy bean oils by high-performance liquid chromatography.

    Science.gov (United States)

    Zonta, F; Stancher, B

    1985-07-19

    A high-performance liquid chromatographic method for determining phylloquinone (vitamin K1) in soy bean oils is described. Resolution of vitamin K1 from interfering peaks of the matrix was obtained after enzymatic digestion, extraction and liquid-solid chromatography on alumina. An isocratic reversed-phase chromatography with UV detection was used in the final stage. The quantitation was carried out by the standard addition method, and the recovery of the whole procedure was 88.2%.

  10. Stationary and through-flow radiochemical detectors in cooperation with high performance liquid chromatography: Application in biochemistry

    International Nuclear Information System (INIS)

    Kehr, J.

    1986-01-01

    A review article is presented containing some original experimental data and discussing the usability of radiochemical detection of labelled compounds using high performance liquid chromatography. The stationary and through-flow types of detection are compared with respect to efficiency, chromatographic zone resolution, usability in biochemical research, and also to the current trends of development of liquid chromatography. (author). 3 figs., 1 tab., 19 refs

  11. High-performance liquid chromatography separation of unsaturated organic compounds by a monolithic silica column embedded with silver nanoparticles.

    Science.gov (United States)

    Zhu, Yang; Morisato, Kei; Hasegawa, George; Moitra, Nirmalya; Kiyomura, Tsutomu; Kurata, Hiroki; Kanamori, Kazuyoshi; Nakanishi, Kazuki

    2015-08-01

    The optimization of a porous structure to ensure good separation performances is always a significant issue in high-performance liquid chromatography column design. Recently we reported the homogeneous embedment of Ag nanoparticles in periodic mesoporous silica monolith and the application of such Ag nanoparticles embedded silica monolith for the high-performance liquid chromatography separation of polyaromatic hydrocarbons. However, the separation performance remains to be improved and the retention mechanism as compared with the Ag ion high-performance liquid chromatography technique still needs to be clarified. In this research, Ag nanoparticles were introduced into a macro/mesoporous silica monolith with optimized pore parameters for high-performance liquid chromatography separations. Baseline separation of benzene, naphthalene, anthracene, and pyrene was achieved with the theoretical plate number for analyte naphthalene as 36,000 m(-1). Its separation function was further extended to cis/trans isomers of aromatic compounds where cis/trans stilbenes were chosen as a benchmark. Good separation of cis/trans-stilbene with separation factor as 7 and theoretical plate number as 76,000 m(-1) for cis-stilbene was obtained. The trans isomer, however, is retained more strongly, which contradicts the long- established retention rule of Ag ion chromatography. Such behavior of Ag nanoparticles embedded in a silica column can be attributed to the differences in the molecular geometric configuration of cis/trans stilbenes. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Analysis of 99mTc-labeled radiopharmaceuticals by high-performance liquid chromatography

    International Nuclear Information System (INIS)

    Muto, Toshio

    1990-01-01

    High-performance liquid chromatography (HPLC) equiped with on line radiometric and optical detectors (i.e. radio-HPLC) have been applied to the radiochemical analysis of commonly-used 99m Tc-radio pharma ceuticals with a view point to check the radiochemical purities of the compounds. Chromatographic conditions were determined by examination of the types of column, mobile phase and pH. An aqueous size-exclusion (Shim-pack Diol-300) and reversed-phase column (Zorbax-ODS) were found to be suitable for 99m Tc-HSA and the other 99m Tc-agents, respectively. The analysis of low molecular weight 99m Tc-agents (e.g. 99m Tc-DTPA, 99m Tc-DMSA, 99m Tc-pyrophosphate, 99m Tc-phytic acid, 99m Tc-MDS, 99m Tc-HMDP) were done by reversed-phaseion pairing chromatography using a optimized mobile phase consisted on a mixture of 50 mM phosphate buffer (pH 7.0) and 2 mM TBA (tetra nbutyl) ammonium hydroxide) in 30 % methanol. The mobile phases for analysis of medium molecular weight 99m Tc-HSA were consisted of a mixture of 50 mM phosphate buffer (ph 7.0) in 30 % methanol, and a mixtures of 1 % SDS (sodium dodecyl sulfonate) in Tris buffer (pH. 7.0), respectively. It was apparent from the radio-chromatograms obtained from these chromatographic conditions, that impurity of 99m TcO 4 was observed in 99m Tc-pyrophosphate, 99m Tc-phytic acid, 99m Tc-MDP, 99m Tc-HMDP, and impurities of 99m Tc-labeled species and 99m TcO 4 , were observed in 99m Tc-HIDA, 99m Tc-HIDA, 99m Tc-HSA. The radiochemical impurities of the 99m Tc-radiopharmaceuticals were ranged between 90 and 100 %. From these results, radio-HPLC has been shown to be suitable method for analysis of 99m Tc-radiopharmaceuticals, with rapidity and excellent precision. (author)

  13. Determination of nucleosides in Cordyceps sinensis and Ganoderma lucidum by high performance liquid chromatography method

    Directory of Open Access Journals (Sweden)

    Masood Shah Khan

    2015-01-01

    Full Text Available Background: Nucleosides are supportive in the regulation and modulation of various physiological processes in body, they acts as precursors in nucleic acid synthesis, enhance immune response, help in absorption of iron and influence the metabolism of fatty acids. Cordyceps sinensis and Ganoderma lucidum are well-known for its use in traditional medicine of China, Nepal and India. They are rich in nucleosides such as adenine, adenosine, cordycepin, etc. Hence, a simple, economic and accurate high-performance liquid chromatography (HPLC analytical method was proposed for determination of adenine and adenosine for the quality control of plants. Materials and Methods: Chromatographic experiments were conducted on YL9100 HPLC system (South Korea. Reversed-phase chromatography was performed on a C18 column with methanol and dihydrogen phosphate as the mobile phase in isocratic elution method at a flow rate of 1.0 mL/min. Detection was carried out at 254 nm, which gives a sharp peak of adenine and adenosine at a retention time of 6.53 ± 0.02 min and 12.41 ± 0.02, respectively. Results and Discussion: Linear regression analysis data for the calibration plot showed a good linear relationship between response and concentration in the range of 25–200 µg/mL for adenosine and 100–800 µg/mL for adenine with regression coefficient of 0.999 and 0.996, respectively. The adenine was found 0.16% and 0.71% w/w in G. lucidum and in C. sinensis, respectively, and adenosine was found to be 0.14% w/w in G. lucidum whereas absent in C. sinensis. Conclusion: The developed HPLC method for the quantification of adenosine and adenine can be used for the quality control and standardization of crude drug and for the different herbal formulations, in which adenine and adenosine are present as major constituents. The wide linearity range, sensitivity, accuracy, and simple mobile phase imply the method is suitable for routine quantification of adenosine and adenine with

  14. Crosslinked polymeric ionic liquids as solid-phase microextraction sorbent coatings for high performance liquid chromatography.

    Science.gov (United States)

    Yu, Honglian; Merib, Josias; Anderson, Jared L

    2016-03-18

    Neat crosslinked polymeric ionic liquid (PIL) sorbent coatings for solid-phase microextraction (SPME) compatible with high-performance liquid chromatography (HPLC) are reported for the first time. Six structurally different PILs were crosslinked to nitinol supports and applied for the determination of select pharmaceutical drugs, phenolics, and insecticides. Sampling conditions including sample solution pH, extraction time, desorption solvent, desorption time, and desorption solvent volume were optimized using design of experiment (DOE). The developed PIL sorbent coatings were stable when performing extractions under acidic pH and remained intact in various organic desorption solvents (i.e., methanol, acetonitrile, acetone). The PIL-based sorbent coating polymerized from the IL monomer 1-vinyl-3-(10-hydroxydecyl) imidazolium chloride [VC10OHIM][Cl] and IL crosslinker 1,12-di(3-vinylbenzylimidazolium) dodecane dichloride [(VBIM)2C12] 2[Cl] exhibited superior extraction performance compared to the other studied PILs. The extraction efficiency of pharmaceutical drugs and phenolics increased when the film thickness of the PIL-based sorbent coating was increased while many insecticides were largely unaffected. Satisfactory analytical performance was obtained with limits of detection (LODs) ranging from 0.2 to 2 μg L(-1) for the target analytes. The accuracy of the analytical method was examined by studying the relative recovery of analytes in real water samples, including tap water and lake water, with recoveries varying from 50.2% to 115.9% and from 48.8% to 116.6%, respectively. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Identification and quantification of amino acids from psoriatic and normal epidermis by high performance liquid chromatography

    International Nuclear Information System (INIS)

    Mahesar, S.M.; Khuhawar, M.Y.

    2010-01-01

    In this study, a modified fluorescence technique high performance liquid chromatography (HPLC) was adapted to separate the amino acids from the hydrolyzed keratin samples. These samples obtained from the epidermal layer of the normal and psoriatic human subjects. The keratin extracts are quantified in gram percentage of the dried skin and the amino acids concentrations are measured in mu g/g, mean retention time (tR), slope value and the coefficient of determination (r2) of each eluted amino acid is calculated. The coefficients of variation for amino acid standards ranged from 0.12% to 0.28%, mean, standard deviation of peak area and coefficients of variation of peak area were calculated. From the normal hydrolysated keratin protein fraction, 12 amino acids were determined and identified as aspartic acid, glutamic acid, asparagines, serine, glutamine, glycine, histidine, citrulline, arganine, fi-alanine, tyrosine, and valine. These amino acids were also determined in psoriatic samples while standard deviations (SD), standard error mean (SEM) and coefficient variation (CV%) of normal and psoriatic samples were also calculated. The higher concentration of amino acids in normal samples against psoriatic samples determined as glutamic acid 92.76+- 16. 83/50. 87+-9.88, glutamine 198.05+-18.74/19.74+-3.74 while higher concentrations of amino acids determined in psoriatic samples against normal samples as asparagines 81. 06+-10+-10.62/29. 98+-3.641; arganine 164.42+-35. 11/46. 14+-46, tyrosine 214.38+-29. 61/59. 64+-8. 82, and valine 169.7+-19.35/128.06+-15.14.1 is concluded that the absolute concentration of amino acids in psoriatic skin indicated a number of variations as compared to normal skin samples. (author)

  16. Evaluation of aminoacids in irradiated beans (Vigna unguiculata (L.) Walp) by high performance liquid chromatography (HPLC)

    Energy Technology Data Exchange (ETDEWEB)

    Lima, Keila S. Cople; Souza, Luciana B.; Coelho, Maysa J.; Lima, Antonio L. Santos; Hernandes, Nilber K. [Instituto Militar de Engenharia (IME), Rio de Janeiro, RJ (Brazil). Secao de Engenharia Nuclear]. E-mail: keila@ime.eb.br; Godoy, Ronoel L.O. [EMBRAPA Agroindustria de Alimentos, Rio de Janeiro, RJ (Brazil)]. E-mail: ronoel@ctaa.embrapa.br

    2007-07-01

    Fradinho-bean (Vigna unguiculata (L.) Walp) is originated from Africa and is known in Brazil as 'caupi', 'corda' or 'macassar'. It is grown in the interior of Northeast Brazil (semi-arid region) and can be found in parts of the North, being one of the most important components of people's diet in those regions. The Northeast area produces around 429,375 ton of fradinho-bean per year. Leguminous plants are very important sources of proteins, vitamins, carbohydrates and minerals. This kind of bean is an excellent source of proteins (around 23- 25% of its nutritional content), being superior to regular beans (Phaseolus vulgaris). The irradiation process is an alternative to avoid post-harvesting losses, without changing the nutritional value of food. This study has the objective to evaluate the effect of different gamma irradiation doses (0.0; 0.5; 1.0; 2.5; 5.0 and 10.0 kGy) on aminoacid content of fradinho-bean by high performance liquid chromatography (HPLC) and the accompanying of the grains during storage time of 6 months. After irradiation, the bean grains went through a milling process in order to make flour for posterior extraction. A liquid chromatographer Waters, model Alliance 2695, with fluorescent detector Waters 2475, having a mobile phase with gradient elution of sodium acetate. acetonitrile and Milli-Q water, was employed. The flux used was 1 mL/min and the injection volume of 10 {mu}L. The column (C 18 150.0 x 3.9 mm) was kept at 36 deg C. The results show that gamma irradiation is a promise process for fradinho bean during conservation storage time of 6 months, until the dose of 10.0 kGy. Even the most radio-sensitive aminoacids like aromatics and basic lateral chains were preserved. (author)

  17. Robust naphthyl methacrylate monolithic column for high performance liquid chromatography of a wide range of solutes.

    Science.gov (United States)

    Jonnada, Murthy; El Rassi, Ziad

    2015-08-28

    An organic monolithic column based on the co-polymerization of 2-naphthyl methacrylate (NAPM) as the functional monomer and trimethylolpropane trimethacrylate (TRIM) as the crosslinker was introduced for high performance reversed-phase liquid chromatography (RPC). The co-polymerization was performed in situ in a stainless steel column of 4.6mm i.d. in the presence of a ternary porogen consisting of 1-dodecanol and cyclohexanol. This monolithic column (referred to as naphthyl methacrylate monolithic column or NMM column) showed high mechanical stability at relatively high mobile phase flow velocity indicating that the column has excellent hydrodynamic characteristics. To characterize the NMM column, different probe molecules including alkyl benzenes, and aniline, benzene, toluene and phenol derivatives were chromatographed on the column and the results in terms of k, selectivity and plate counts were compared to those obtained on an octadecyl silica (ODS) column in order to assess the presence of π-π and hydrophobic interactions on the NMM column under otherwise the same elution conditions. The NMM column offered additional π-π interactions with aromatic molecules in addition to hydrophobic interactions under RPC elution conditions. Run-to-run and column-to-column reproducibility of solute k values were evaluated, and percent relative standard deviation of column using shallow (30min at 1.0mL/min), steep (10min at 1.0mL/min) and ultra steep (1min at 3.0mL/min) linear gradient elution at increasing ACN concentration in the mobile phase using a 10cm×4.6mm i.d. column in case of shallow and steep linear gradients and a 3cm×4.6mm i.d. column for ultra steep linear gradient. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Determination of puerarin in rat plasma using PEGylated magnetic carbon nanotubes by high performance liquid chromatography.

    Science.gov (United States)

    Yu, Panfeng; Wang, Qi; Ma, Hongwei; Wu, Ji; Shen, Shun

    2014-05-15

    This paper described a novel application of PEGylated magnetic carbon nanotubes as solid-phase extraction nanosorbents for the determination of puerarin in rat plasma by high performance liquid chromatography (HPLC). A solvothermal method was employed for the synthesis of monodisperse magnetites anchored onto multi-walled carbon nanotubes (MWCNTs@Fe3O4). In order to enhance the water solubility of MWCNTs@Fe3O4 that ensured sufficient contact between nanosorbents and analytes in the sampling procedure, the obtained nanomaterials were further noncovalently functionalized using a phospholipids-polyethylene glycol (DSPE-PEG). The PEGylated MWCNTs@Fe3O4 nanomaterials had an extremely large surface area and exhibit a strong interaction capability for puerarin with π-π stacking interactions. The captured puerarin/nanosorbents were easily isolated from the plasma by placing a magnet, and desorbed by acetonitrile. The experimental variables affecting the extraction efficiency were investigated. The calibration curve of puerarin was linear from 0.01 to 20 μg/ml, and the limit of detection was 0.005 μg/ml. The precisions ranged from 2.7% to 3.5% for within-day measurement, and for between-day variation was in the range of 3.1-5.9%. The method recoveries were acquired from 95.2% to 98.0%. Moreover, the analytical performance obtained by PEGylated magnetic MWCNTs was also compared with that of magnetic MWCNTs. All results showed that our proposed method was an excellent alternative for the analysis of puerarin in rat plasma. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Characterization of detergent-solubilized sarcoplasmic reticulum Ca2+-ATPase by high-performance liquid chromatography

    International Nuclear Information System (INIS)

    Andersen, J.P.; Vilsen, B.; Nielsen, H.; Moller, J.V.

    1986-01-01

    Sarcoplasmic reticulum Ca 2+ -ATPase solubilized by the nonionic detergent octaethylene glycol monododecyl ether was studied by molecular sieve high-performance liquid chromatography (HPLC) and analytical ultracentrifugation. Significant irreversible aggregation of soluble Ca 2+ -ATPase occurred within a few hours in the presence of ≤ 50 μM Ca 2+ . The aggregates were inactive and were primarily held together by hydrophobic forces. In the absence of reducing agent, secondary formation of disulfide bonds occurred. The stability of the inactive dimer upon dilution permitted unambiguous assignment of its elution position and sedimentation coefficient. At high 45 Ca 2+ concentration (500 μM), monomeric Ca 2+ -ATPase was stable for several house. Reversible self-association induced by variation in protein, detergent, and lipid concentrations was studied by large-zone HPLC. The association constant for dimerization of active Ca 2+ -ATPase was found to be 10 5 -10 6 M -1 depending on the detergent concentration. More detergent was bound to monomeric than to dimeric Ca 2+ -ATPase, even above the critical micellar concentration of the detergent. Binding of Ca 2+ and 48 V vanadate as well as ATP-dependent phosphorylation was studied in monomeric and in reversibly associated dimeric preparations. In both forms, two high-affinity Ca 2+ binding sites per phosphorylation site existed. The delipidated monomer purified by HPLC was able to form ADP-insensitive phosphoenzyme and to bind ATP and vanadate simultaneously. The results suggest that formation of Ca 2+ -ATPase oligomers in the membrane is governed by nonspecific forces (low affinity) and that each polypeptide chain constitutes a functional unit

  20. Quantification of free formaldehyde in carrageenan and processed Eucheuma seaweed using high-performance liquid chromatography.

    Science.gov (United States)

    Hornshøj, Bettina Høj; Kobbelgaard, Sara; Blakemore, William R; Stapelfeldt, Henrik; Bixler, Harris J; Klinger, Markus

    2015-01-01

    In 2010 the European Commission placed a limit on the amount of free formaldehyde in carrageenan and processed Eucheuma seaweed (PES) of 5 mg kg(-1). Formaldehyde is not used in carrageenan and PES processing and accordingly one would not expect free formaldehyde to be present in carrageenan and PES. However, surprisingly high levels up to 10 mg kg(-1) have been found using the generally accepted AOAC and Hach tests. These findings are, per proposed reaction pathways, likely due to the formation of formaldehyde when sulphated galactose, the backbone of carrageenan, is hydrolysed with the strong acid used in these conventional tests. In order to minimise the risk of false-positives, which may lead to regulatory non-compliance, a new high-performance liquid chromatography (HPLC) method has been developed. Initially, carrageenan or PES is extracted with 2-propanol and subsequently reacted with 2,4-dinitrophenylhydrazine (DNPH) to form the chromophore formaldehyde-DNPH, which is finally quantified by reversed-phase HPLC with ultraviolet light detection at 355 nm. This method has been found to have a limit of detection of 0.05 mg kg(-1) and a limit of quantification of 0.2 mg kg(-1). Recoveries from samples spiked with known quantities of formaldehyde were 95-107%. Using this more specific technique, 20 samples of carrageenan and PES were tested for formaldehyde. Only one sample had a detectable content of formaldehyde (0.40 mg kg(-1)), thus demonstrating that the formaldehyde content of commercial carrageenan and PES products are well below the European Commission maximum limit of 5 mg kg(-1).

  1. Quantitative determination of insulin entrapment efficiency in triblock copolymeric nanoparticles by high-performance liquid chromatography.

    Science.gov (United States)

    Xu, Xiongliang; Fu, Yao; Hu, Haiyan; Duan, Yourong; Zhang, Zhirong

    2006-04-11

    A rapid and effective isocratic chromatographic procedure was described in this paper for the determination of insulin entrapment efficiency (EE) in triblock copolymeric nanoparticles using reversed-phase high-performance liquid chromatography (RP-HPLC) with an ultraviolet/visible detector at low flow rate. The method has been developed on a Shimadzu Shim-pack VP-ODS column (150 mm x 4.6 mm, 5 microm, Chiyoda-Ku, Tokyo, Japan) using a mixture of 0.2 M sodium sulfate anhydrous solution adjusted to pH 2.3 with phosphoric acid and acetonitrile (73:27, v/v) as mobile phase at the flow rate of 0.8 ml min(-1) and a 214 nm detection. The method was validated in terms of selectivity, linearity, precision, accuracy, solution stability, limit of detection (LOD) and limit of quantification (LOQ). The calibration curve was linear in the concentration range of 2.0-500.0 microg ml(-1), and the limits of detection and quantitation were 8 and 20 ng, respectively. The mean recovery of insulin from spiked samples, in a concentration range of 8-100 microg ml(-1), was 98.96% (R.S.D.= 2.51%, n = 9). The intra- and inter-assay coefficients of variation were less than 2.24%. The proposed method has the advantages of simple pretreatment, rapid isolation, high specificity and precision, which can be used for direct analysis of insulin in commercially available raw materials, formulations of nanoparticles, and drug release as well as stability studies.

  2. Histone fractionation by high-performance liquid chromatography on cyanoalkylsilane (CN) reverse-phase columns

    International Nuclear Information System (INIS)

    Gurley, L.R.; Prentice, D.A.; Valdez, J.G.; Spall, W.D.

    1983-01-01

    Previous work described conditions for the rapid fractionation of histones by high-performance liquid chromatography (HPLC) using a reverse-phase μBondapak C 18 column. That procedure resolved the major classes of histones with one exception: the more hydrophobic H2A variant, (MHP)H2A, was not resolved from the H4 histone class. This report extends that work describing experiments using a μBondapak CN column which better resolves the classes of histones from each other including the resolution of (MHP)H2A from the H4. In addition, the less hydrophobic H2A variant, (LHP)H2A, is partially resolved from the (MHP)H2A, and the less hydrophobic H3 variant, (LHP)H3, is resolved from the more hydrophobic H3 variant, (MHP)H3. Lower trifluoroacetic acid (TFA) concentrations (0.1%) in the eluting water/acetonitrile solvent were used with the CN column than were used with the C 18 column which increased the sensitivity of histone detection by ultraviolet absorption at 206 nm. Greater than 95% of the total [ 3 H]lysine-labeled protein applied to the CN column was eluted from the column. Contaminating nonhistone proteins were found to chromatograph in the region of histone elution. These were greatly reduced by isolating nuclei prior to histone preparation. The fractionation of the histones appears to be based on the hydrophobic properties of the proteins. The histone fractions (identified by their electrophoretic mobilities) were eluted from the CN column in the following order: H1, H2B, (LHP)H2A, (MHP)H2A, H4, (LHP)H3, and (MHP)H3. Phosphorylated and acetylated histone species were not resolved from their unmodified parental species

  3. Analysis of I-125 IMP and its metabolites using a high performance liquid chromatography

    International Nuclear Information System (INIS)

    Satoh, Motohiro; Ishikawa, Nobuyoshi; Takeda, Tohoru; Jin, Wu; Kuramoto, Kenmei; Itai, Yuji; Yoshizawa, Takashi; Nakajima, Kotaro.

    1991-01-01

    The biodistribution of N-isopropyl-p-[I-123]iodoamphetamine (IMP) and its metabolites was examined in rabbits and Mongolian gerbils. Arterial sampling was performed at one, 5, 15, and 30 min, and one, 3, and 6 hr after bolus iv injection of IMP for the hemodynamic investigation. Similarly, the cerebral hemisphere, lung, liver, and blood samples were collected at 15 min and 3 hr for analyzing IMP metabolites. Activity count in blood was gradually increased from 15 min to 3 hr after iv injection, and thereafter decreased. Relative fraction of IMP in plasma was gradually increased to a plateau value of 80% at one hr. Octanol extraction ratio was decreased to 24.3% at 3 hr, although it was 100% immediately after iv injection. Early (15 min) and delayed (3 hr) analysis using high performance liquid chromatography (HPLC) revealed p-iodoamphetamine (PIA) and p-iodobenzoic acid (PIB) as major metabolites of IMP. Although IMP accounted for the majority on both early and delayed HPLC, the quantity of PIA in the normal hemisphere and lung was significantly increased on delayed HPLC, compared to early HPLC. For the liver, the quantities of both PIA and PIB were larger than IMP on both early and delayed HPLC. The proportion of metabolites also became greater in whole blood than IMP on delayed HPLC. Early HPLC reveald no significant difference in composition of IMP, PIA, and PIB between the normal and ischemic hemispheres. Delayed HPLC revealed a greater proportion of PIA in the ischemic than the normal hemisphere, but this was not statistically significant. (N.K.)

  4. Simultaneous determination of caffeine, theobromine, and theophylline by high-performance liquid chromatography.

    Science.gov (United States)

    Bispo, Marcia S; Veloso, Márcia Cristina C; Pinheiro, Heloísa Lúcia C; De Oliveira, Rodolfo F S; Reis, José Oscar N; De Andrade, Jailson B

    2002-01-01

    This work relates the development of an analytical methodology to simultaneously determine three methylxanthines (caffeine, theobromine, and theophylline) in beverages and urine samples based on reversed-phase high-performance liquid chromatography. Separation is made with a Bondesil C18 column using methanol-water-acetic acid or ethanol-water-acetic acid (20:75:5, v/v/v) as the mobile phase at 0.7 mL/min. Identification is made by absorbance detection at 273 nm. Under optimized conditions, the detection limit of the HPLC method is 0.1 pg/mL for all three methylxanthines. This method is applied to urine and to 25 different beverage samples, which included coffee, tea, chocolate, and coconut water. The concentration ranges determined in the beverages and urine are: < 0.1 pg/mL to 350 microg/mL and 3.21 microg/mL to 71.2 microg/mL for caffeine; < 0.1 pg/mL to 32 microg mL and < 0.1 pg/mL to 13.2 microg/mL for theobromine; < 0.1 pg/mL to 47 microg/mL and < 0.1 pg/mL to 66.3 microg/mL for theophylline. The method proposed in this study is rapid and suitable for the simultaneous quantitation of methylxanthines in beverages and human urine samples and requires no extraction step or derivatization.

  5. Flavonoids from the flowers of Impatiens glandulifera Royle isolated by high performance countercurrent chromatography.

    Science.gov (United States)

    Vieira, Mariana N; Winterhalter, Peter; Jerz, Gerold

    2016-01-01

    Impatiens glandulifera Royle (Balsaminaceae) is an annual herb from the Himalaya region, currently widespread along European river systems and one of the most important neophyte invading plants in Germany. Exploring the effects of allelopathic plant chemicals is important for the understanding of its ecological impacts in the process of suppression of indigenous plant species. To investigate the chemical composition of Impatiens glandulifera flowers (IGFs) using high performance countercurrent chromatography (HPCCC). The flowers of Impatiens glandulifera were manually separated and extracted with ethanol. LC-ESI-MS/MS was used to characterise the crude extract of IGF. The various flavonoids detected were isolated by HPCCC using of methyl tert-butyl ether-acetonitrile-water (2:2:3, v/v/v). The combination of the data provided by preparative ESI-MS/MS metabolite profiling, LC-ESI-MS/MS, UV-vis and 1D/2D-NMR spectroscopic analysis was used to elucidate the structures of the isolated compounds. HPCCC runs led to the direct isolation of pure dihydromyricetin (ampelopsin), eriodictyol-7-O-glucoside, kaempferol-3-O-glucoside (astragalin) and kaempferol-3-O-6"-malonyl-glucoside, as well as the pre-purification of kaempferol-3-O-rhamno-rhamnosyldiglucoside, quercetin-3-O-galactoside (hyperoside), quercetin and kaempferol in a single step. This is the first report on the flavonoid composition of the species Impatiens glandulifera. The developed protocol was successfully used to isolate the main flavonoids from the crude extract of IGFs. This combined HPCCC and HPLC procedure could be applied to the fast fractionation and recovery of flavonoid derivatives of other plant extracts. Copyright © 2016 John Wiley & Sons, Ltd.

  6. Radiotracer properties determined by high performance liquid chromatography: a potential tool for brain radiotracer discovery

    International Nuclear Information System (INIS)

    Tavares, Adriana Alexandre S.; Lewsey, James; Dewar, Deborah; Pimlott, Sally L.

    2012-01-01

    Introduction: Previously, development of novel brain radiotracers has largely relied on simple screening tools. Improved selection methods at the early stages of radiotracer discovery and an increased understanding of the relationships between in vitro physicochemical and in vivo radiotracer properties are needed. We investigated if high performance liquid chromatography (HPLC) methodologies could provide criteria for lead candidate selection by comparing HPLC measurements with radiotracer properties in humans. Methods: Ten molecules, previously used as radiotracers in humans, were analysed to obtain the following measures: partition coefficient (Log P); permeability (P m ); percentage of plasma protein binding (%PPB); and membrane partition coefficient (K m ). Relationships between brain entry measurements (Log P, P m and %PPB) and in vivo brain percentage injected dose (%ID); and K m and specific binding in vivo (BP ND ) were investigated. Log P values obtained using in silico packages and flask methods were compared with Log P values obtained using HPLC. Results: The modelled associations with %ID were stronger for %PPB (r 2 =0.65) and P m (r 2 =0.77) than for Log P (r 2 =0.47) while 86% of BP ND variance was explained by K m . Log P values were variable dependant on the methodology used. Conclusions: Log P should not be relied upon as a predictor of blood-brain barrier penetration during brain radiotracer discovery. HPLC measurements of permeability, %PPB and membrane interactions may be potentially useful in predicting in vivo performance and hence allow evaluation and ranking of compound libraries for the selection of lead radiotracer candidates at early stages of radiotracer discovery.

  7. Development and validation of reverse phase high performance liquid chromatography for citral analysis from essential oils.

    Science.gov (United States)

    Gaonkar, Roopa; Yallappa, S; Dhananjaya, B L; Hegde, Gurumurthy

    2016-11-15

    Citral is a widely used monoterpene aldehyde in aromatherapy, food and pesticide industries. A new validated reverse phase high performance liquid chromatography (RP - HPLC) procedure for the detection and quantification of cis-trans isomers of citral was developed. The RP-HPLC analysis was carried out using Enable C - 18G column (250×4.6mm, 5μ), with acetonitrile and water (70: 30) mobile phase in isocratic mode at 1mL/min flow. A photodiode array (PDA) detector was set at 233nm for the detection of citral. The method showed linearity, selectivity and accuracy for citral in the range of 3-100μg/mL. In order to compare the new RP-HPLC method with the available methods, one of the commercially available essential oil from Cymbopogon flexuosus was analyzed using new RP-HPLC method and the same was analyzed using GC-MS for the comparison of the method for the detection of citral. The GC-MS analysis was done using mass selective detector (MSD) showed citral content to be of 72.76%; wherein the new method showed to contain that same at 74.98%. To prove the application of the new method, essential oils were extracted from lemongrass, lemon leaves and mosambi peels by steam distillation. The citral content present in the essential and also in the condensate was analyzed. The method was found to be suitable for the analysis of citral in essential oils and water based citral formulations with a very good resolution of its components geranial and neral. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Improved method for the determination of the cortisol production rate using high-performance liquid chromatography and liquid scintillation counting

    NARCIS (Netherlands)

    van Ingen, H. E.; Endert, E.

    1988-01-01

    Two new methods for the determination of the cortisol production rate using reversed-phase high-performance liquid chromatography are described. One uses ultraviolet detection at 205 nm, the other on-line post-column derivatization with benzamidine, followed by fluorimetric detection. The specific

  9. Simultaneous determination of estrogens and progestogens in honey using high performance liquid chromatography-tandem mass spectrometry

    Science.gov (United States)

    This work describes the development and validation of a method for the simultaneous determination of 13 estrogens and progestogens in honey by high performance liquid chromatography-tandem mass spectrometry. The target compounds were preconcentrated by solid phase extraction. Pretreatment variables ...

  10. Analysis of aqueous humour in uveitis by high performance liquid chromatography and sodium dodecyl sulphate-polyacrylamide gel electrophoresis

    NARCIS (Netherlands)

    Murray, P. I.; Hoekzema, R.; Luyendijk, L.; Kijlstra, A.

    1992-01-01

    Aqueous humour from patients with Fuchs' heterochromic cyclitis (FHC) and other types of uveitis was analysed by high performance liquid chromatography (HPLC) and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Using HPLC, the number of peaks and their respective elution times

  11. Theoretical aspects of gradient reversed-phase high performance liquid chromatography of styrene-butylacrylate block copolymers

    NARCIS (Netherlands)

    Kolarova, L.; Jandera, P.; Vonk, E.C.; Claessens, H.A.

    2004-01-01

    Butylacrylate – styrene co-polymers prepared by atom transfer radical polymeratization were separated on an octadecyl silica column by gradient elution with tetrahydrofuran in water, up to the molar masses 10,000. In reversed-phase high performance liquid chromatography (RP-HPLC), the retention of

  12. Standardization of heparins by means of high performance liquid chromatography equipped with a low angle laser light scattering detector

    NARCIS (Netherlands)

    Hennink, W.E.; van den Berg, J.W.A.; Feijen, Jan

    1987-01-01

    This study shows that HPLC-LALLS (high performance liquid chromatography with a light-scattering detector) is a convenient and reliable method for the characterization of standard heparin samples, provided that polyelectrolyte artefacts are suppressed by a suitable dialysis procedure. The method has

  13. NONLINEAR-REGRESSION METHODS FOR MODELING OF HETEROSCEDASTIC RETENTION DATA IN REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

    NARCIS (Netherlands)

    HENDRIKS, MMWB; COENEGRACHT, PMJ; DOORNBOS, DA

    1994-01-01

    New models have been developed that accurately describe the response surfaces of capacity factors that are a function of changes in the pH and the fraction of organic modifier in reversed-phase high-performance liquid chromatography (RP-HPLC). The purpose of this article is to illustrate one of the

  14. High-performance liquid chromatography-mass spectrometry-based acetylcholinesterase assay for the screening of inhibitors in natural extracts

    NARCIS (Netherlands)

    de Jong, C.F.; Derks, R.J.E.; Bruyneel, B.; Niessen, W.M.A.; Irth, H.

    2006-01-01

    The present paper describes a High-performance liquid chromatography-mass spectrometry (LC-MS) methodology for the screening of acetylcholinesterase (AChE) inhibitors in natural extracts. AChE activity of sample components is monitored by a post-column biochemical assay that is based on the

  15. Separation and Quantitation of Polyamines in Plant Tissue by High Performance Liquid Chromatography of Their Dansyl Derivatives

    Science.gov (United States)

    Smith, Mary A.; Davies, Peter J.

    1985-01-01

    High performance liquid chromatography in combination with fluorescence spectrophotometry can be used to separate and quantitate polyamines (putrescine, cadaverine, spermidine, spermine), prepared as their dansyl derivatives, from plant tissue. The procedure gives sensitive and consistent results for polyamine determinations in plant tissue. In a standard mixture, the minimal detection level was less than 1 picomole of polyamines. PMID:16664216

  16. Quantitation of bilirubin conjugates with high-performance liquid chromatography in patients with low total serum bilirubin levels

    NARCIS (Netherlands)

    Jansen, P. L.; Cuypers, H. T.; Peters, W. H.

    1984-01-01

    Bilirubin mono- and diconjugates were determined by alkaline methanolysis and high-performance liquid chromatography (HPLC) in serum from patients with metastatic liver disease and liver cirrhosis. Conjugates could be detected and quantitated at normal or low total bilirubin levels. Comparison with

  17. Determination of methyldibromoglutaronitrile in cosmetic products by high-performance liquid chromatography with electrochemical detection. Method validation

    DEFF Research Database (Denmark)

    Rastogi, Suresh Chandra; Zachariae, Claus; Johansen, Jeanne D

    2004-01-01

    An increased frequency of contact allergy to methyldibromoglutaronitrile (MDBGN), a commonly used preservative in cosmetics and other consumer products, has been reported in recent years. A high-performance liquid chromatography (HPLC) method for the determination of MDBGN in cosmetic products ha...

  18. Determination of methyldibromoglutaronitrile in cosmetic products by high-performance liquid chromatography with electrochemical detection. Method validation

    DEFF Research Database (Denmark)

    Rastogi, Suresh Chandra; Zachariae, Claus; Johansen, Jeanne D

    2004-01-01

    An increased frequency of contact allergy to methyldibromoglutaronitrile (MDBGN), a commonly used preservative in cosmetics and other consumer products, has been reported in recent years. A high-performance liquid chromatography (HPLC) method for the determination of MDBGN in cosmetic products has...

  19. Simultaneous Determination of Caffeine and Vitamin B6 in Energy Drinks by High-Performance Liquid Chromatography (HPLC)

    Science.gov (United States)

    Leacock, Rachel E.; Stankus, John J.; Davis, Julian M.

    2011-01-01

    A high-performance liquid chromatography experiment to determine the concentration of caffeine and vitamin B6 in sports energy drinks has been developed. This laboratory activity, which is appropriate for an upper-level instrumental analysis course, illustrates the standard addition method and simultaneous determination of two species. (Contains 1…

  20. Constructing a LabVIEW-Controlled High-Performance Liquid Chromatography (HPLC) System: An Undergraduate Instrumental Methods Exercise

    Science.gov (United States)

    Smith, Eugene T.; Hill, Marc

    2011-01-01

    In this laboratory exercise, students develop a LabVIEW-controlled high-performance liquid chromatography system utilizing a data acquisition device, two pumps, a detector, and fraction collector. The programming experience involves a variety of methods for interface communication, including serial control, analog-to-digital conversion, and…

  1. Analysis of native human plasma proteins and haemoglobin for the presence of bityrosine by high-performance liquid chromatography

    DEFF Research Database (Denmark)

    Daneshvar, B; Frandsen, H; Dragsted, L O

    1997-01-01

    fluorescent substance, bityrosine. High-performance liquid chromatography (HPLC) analysis of acid hydrolyzed serum albumin after oxidation with peroxidase/H2O2 or with Cu++/H2O2 showed that bityrosine had been formed whereas oxidation of this protein with Fe(III)/ascorbate did not result in the formation...

  2. The Determination of Polyethylene Glycol in Untreated Urine Samples by High Performance Liquid Chromatography for Intestinal Permeability Studies

    DEFF Research Database (Denmark)

    Larsen, Elfinn; Pedersen, Walther Batsberg; Philipsen, E.

    1985-01-01

    Polyethylene glycol in urine samples has been investigated by high performance liquid chromatography. The molecular weights ranged from 634 to 1338. The urine samples were applied to the chromatographic system without any pre-treatment. For samples with a concentration of 0.2% polyethylene glycol...

  3. Identification and quantification of cannabinoids in Cannabis sativa L. plants by high performance liquid chromatography-mass spectrometry

    NARCIS (Netherlands)

    Aizpurua-Olaizola, Oier; Omar, Jone; Navarro, Patricia; Olivares, Maitane; Etxebarria, Nestor; Usobiaga, Aresatz

    2014-01-01

    High performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) has been successfully applied to cannabis plant extracts in order to identify cannabinoid compounds after their quantitative isolation by means of supercritical fluid extraction (SFE). MS conditions were optimized by means

  4. Screening anti-tumor compounds from Ligusticum wallichii using cell membrane chromatography combined with high-performance liquid chromatography and mass spectrometry.

    Science.gov (United States)

    Zhang, Tao; Ding, Yuanyuan; An, Hongli; Feng, Liuxin; Wang, Sicen

    2015-07-14

    Tyrosine 367 Cysteine-fibroblast growth factor receptor 4 cell membrane chromatography combined with high-performance liquid chromatography and mass spectrometry was developed. Tyrosine 367 Cysteine-HEK293 cells were used as cell membrane stationary phase. Specificity and reproducibility of the cell membrane chromatography was evaluated using 1-tert-butyl-3-{2-[4-(diethylamino)butylamino]-6-(3,5-dimethoxyphenyl)pyrido[2,3-d]pyrimidin-7-yl}urea, Nimodipine and dexamethasone acetate. Then, anti-tumor components acting on Tyrosine 367 Cysteine-fibroblast growth factor receptor 4 were screened and identified from extracts of Ligusticum wallichii. Components from the extract were retained on the cell membrane chromatographic column. The retained fraction was directly eluted into high-performance liquid chromatography with mass spectrometry system for separation and identification. Finally, Levistolide A was identified as an active component from Ligusticum wallichii extracts. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide-formazan colorimetric assay revealed that Levistolide A inhibits proliferation of overexpressing the mutated receptor cells with dose-dependent manner. Phosphorylation of fibroblast growth factor receptor 4 was also decrease under Levistolide A treatment. Flex dock simulation verified that Levistolide A could bind with the tyrosine kinase domain of fibroblast growth factor receptor 4. Therefore, Levistolide A screened by the cell membrane chromatography combined with high-performance liquid chromatography and mass spectrometry can arrest cell growth. In conclusion, the two-dimensional high-performance liquid chromatography method can screen and identify potential anti-tumor ingredients which specifically act on the tyrosine kinase domain of the mutated fibroblast growth factor receptor 4. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  5. Rapid determination of human globin chains using reversed-phase high-performance liquid chromatography.

    Science.gov (United States)

    Wan, Jun-Hui; Tian, Pei-Ling; Luo, Wei-Hao; Wu, Bing-Yi; Xiong, Fu; Zhou, Wan-Jun; Wei, Xiang-Cai; Xu, Xiang-Min

    2012-07-15

    Reversed-phase high-performance liquid chromatography (RP-HPLC) of human globin chains is an important tool for detecting thalassemias and hemoglobin variants. The challenges of this method that limit its clinical application are a long analytical time and complex sample preparation. The aim of this study was to establish a simple, rapid and high-resolution RP-HPLC method for the separation of globin chains in human blood. Red blood cells from newborns and adults were diluted in deionized water and injected directly onto a micro-jupiter C18 reversed-phase column (250 mm × 4.6 mm) with UV detection at 280 nm. Under the conditions of varying pH or the HPLC gradient, the globin chains (pre-β, β, δ, α, (G)γ and (A)γ) were denatured and separated from the heme groups in 12 min with a retention time coefficient of variation (CV) ranging from 0.11 to 1.29% and a peak area CV between 0.32% and 4.86%. Significant differences (P<0.05) among three groups (normal, Hb H and β thalassemia) were found in the area ratio of α/pre-β+β applying the rapid elution procedure, while P≥0.05 was obtained between the normal and α thalassemia silent/trait group. Based on the ANOVA results, receiver operating characteristic (ROC) curve analysis of the δ/β and α/pre-β+β area ratios showed a sensitivity of 100.0%, and a specificity of 100.0% for indicating β thalassemia carriers, and a sensitivity of 96.6% and a specificity of 89.6% for the prediction of hemoglobin H (Hb H) disease. The proposed cut-off was 0.026 of δ/β for β thalassemia carriers and 0.626 of α/pre-β+β for Hb H disease. In addition, abnormal hemoglobin hemoglobin E (Hb E) and Hb Westmead (Hb WS) were successfully identified using this RP-HPLC method. Our experience in developing this RP-HPLC method for the rapid separation of human globin chains could be of use for similar work. Copyright © 2012 Elsevier B.V. All rights reserved.

  6. Development and validation of methodology for technetium-99m radiopharmaceuticals using high performance liquid chromatography (HPLC)

    International Nuclear Information System (INIS)

    Almeida, Erika Vieira de

    2009-01-01

    Radiopharmaceuticals are compounds, with no pharmacological action, which have a radioisotope in their composition and are used in Nuclear Medicine for diagnosis and therapy of several diseases. In this work, the development and validation of an analytical method for 99 mTc-HSA, 99 mTc-EC, 99 mTc-ECD and 99 mTc-Sestamibi radiopharmaceuticals and for some raw materials were carried out by high performance liquid chromatography (HPLC). The analyses were performed in a Shimadzu HPLC equipment, LC-20AT Prominence model. Some impurities were identified by the addition of a reference standard substance. Validation of the method was carried out according to the criteria defined in RE n. 899/2003 of the National Sanitary Agency (ANVISA). The results for robustness of the method showed that it is necessary to control flow rate conditions, sample volume, pH of the mobile phase and temperature of the oven. The analytical curves were linear in the concentration ranges, with linear correlation coefficients (r 2 ) above 0.9995. The results for precision, accuracy and recovery showed values in the range of 0.07-4.78%, 95.38-106.50% and 94.40-100.95%, respectively. The detection limits and quantification limits varied from 0.27 to 5.77 μg mL -1 and 0.90 to 19.23 μg mL -1 , respectively. The values for HAS, EC, ECD and MIBI in the lyophilized reagents were 8.95; 0.485; 0.986 and 0.974 mg L-1, respectively. The mean radiochemical purity for 99 mTc-HSA, 99 mTc-EC, 99 mTc-ECD and 99 mTc-Sestamibi was (97.28 ± 0.09)%, (98.96 ± 0.03)%, (98.96 ± 0.03)% and (98.07 ± 0.01)%, respectively. All the parameters recommended by ANVISA were evaluated and the results are below the established limits. (author)

  7. High-performance liquid chromatography electrospray mass spectrometry as a method in proteomic research

    International Nuclear Information System (INIS)

    Walcher, W.

    2003-06-01

    During the sequencing of the human genome it became clear, that a lot of human diseases and/or malfunctions don't base on genomic information, but on differences at the protein level. Therefore biochemistry, biology and medicine are faced to various novel problems where new and authentic analysis methods are needed. Miniaturized chromatographic separation methods are frequently the methods of choice for the separation of peptides and proteins, when the amount of sample is limited. Monolithic capillary columns were prepared by copolymerization of styrene and divinylbenzene in the presence of a suitable porogen mixture of 1-decanol and tetrahydrofuran. The synthesized columns enabled the highly efficient separation of peptides and proteins by reversed-phase high-performance liquid chromatography (RP-HPLC) with peak capacities of 80 and more in 10 minutes. By the hyphenation of RP-HPLC to electrospray mass spectrometry (ESI-MS) the potential of the analysis method was even extended. The monolithic column technology was further miniaturized from 200 μm to 100 and 50 μm inner diameters to improve detection limits by RP-HPLC-ESI-MS. After the optimization of the RP-HPLC-ESI-MS method, the ion source and the ion transfer optics of an ion trap mass spectrometer (LCQ classic, Thermofinnigan) have been advanced for protein and peptide analysis. The improved RP-HPLC-ESI-MS system was subsequently applied to the detection of posttranslational protein modifications at the example of the nitration of bovine serum albumin (BSA). The results of the RP-HPLC-ESI-MS analysis were found to be highly reproducible, which enabled the determination of nitration degrees for different tyrosine residues in the protein sequence. Y 1 60, Y 4 96 and Y 3 52 or Y369 were found to be the predominant positions of protein nitration in BSA. At last light harvesting proteins from the photosystem II (PSII) of higher plants have been analyzed by RP-HPLC-ESI-MS and RP-HPLC-ESI-MSMS. Beside the

  8. Recent advances in ultra-high performance liquid chromatography for the analysis of traditional chinese medicine

    Science.gov (United States)

    Traditional Chinese medicines (TCMs) have been widely used for the prevention and treatment of various diseases for thousands of years in China. Ultra Performance Liquid Chromatography (UHPLC) is a relatively new technique offering new possibilities in liquid chromatography. This paper reviews recen...

  9. Analysis of the Constituents in “Zhu She Yong Xue Shuan Tong” by Ultra High Performance Liquid Chromatography with Quadrupole Time-of-Flight Mass Spectrometry Combined with Preparative High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Lin-Lin Wang

    2015-11-01

    Full Text Available “Zhu She Yong Xue Shuan Tong” lyophilized powder (ZSYXST, consists of a series of saponins extracted from Panax notoginseng, which has been widely used in China for the treatment of strokes. In this study, an ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS combined with preparative high performance liquid chromatography (PHPLC method was developed to rapidly identify both major and minor saponins in ZSYXST. Some high content components were removed through PHPLC in order to increase the sensitivity of the trace saponins. Then, specific characteristic fragment ions in both positive and negative mode were utilized to determine the types of aglycone, saccharide, as well as the saccharide chain linkages. As a result, 94 saponins, including 20 pairs of isomers and ten new compounds, which could represent higher than 98% components in ZSYXST, were identified or tentatively identified in commercial ZSYXST samples.

  10. High Performance Liquid Chromatography Determination of Urinary Hippuric Acid and Benzoic Acid as Indices for Glue Sniffer Urine

    OpenAIRE

    Abdul Rahim Yacob; Mohamad Raizul Zinalibdin

    2010-01-01

    A simple method for the simultaneous determination of hippuric acid and benzoic acid in urine using reversed-phase high performance liquid chromatography was described. Chromatography was performed on a Nova-Pak C18 (3.9 x 150 mm) column with a mobile phase of mixed solution methanol: water: acetic acid (20:80:0.2) and UV detection at 254 nm. The calibration curve was linear within concentration range at 0.125 to 6.0 mg/ml of hippuric acid and benzoic acid. The recovery, ...

  11. Determination of Aspartame, Caffeine, Saccharin, and Benzoic Acid in Beverages by High Performance Liquid Chromatography.

    Science.gov (United States)

    Delaney, Michael F.; And Others

    1985-01-01

    Describes a simple and reliable new quantitative analysis experiment using liquid chromatography for the determinaiton of caffeine, saccharin, and sodium benzoate in beverages. Background information, procedures used, and typical results obtained are provided. (JN)

  12. Instrumental liquid chromatography: a practical manual on high-performance liquid chromatographic methods

    National Research Council Canada - National Science Library

    Parris, N. A

    1976-01-01

    Available texts on liquid chromatography have tended to emphasize the developments in the theoretical understanding of the technique and methodology or to list numerous applications, complete with experimental details...

  13. Detection of argan oil adulteration with vegetable oils by high-performance liquid chromatography-evaporative light scattering detection.

    Science.gov (United States)

    Salghi, Rachid; Armbruster, Wolfgang; Schwack, Wolfgang

    2014-06-15

    Triacylglycerol profiles were selected as indicator of adulteration of argan oils to carry out a rapid screening of samples for the evaluation of authenticity. Triacylglycerols were separated by high-performance liquid chromatography-evaporative light scattering detection. Different peak area ratios were defined to sensitively detect adulteration of argan oil with vegetable oils such as sunflower, soy bean, and olive oil up to the level of 5%. Based on four reference argan oils, mean limits of detection and quantitation were calculated to approximately 0.4% and 1.3%, respectively. Additionally, 19 more argan oil reference samples were analysed by high-performance liquid chromatography-refractive index detection, resulting in highly comparative results. The overall strategy demonstrated a good applicability in practise, and hence a high potential to be transferred to routine laboratories. Copyright © 2013 Elsevier Ltd. All rights reserved.

  14. Quantification of LSD in illicit samples by high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Pablo Alves Marinho

    2010-12-01

    Full Text Available In the present study, a method using high performance liquid chromatography to quantify LSD, in blotter papers seized in Minas Gerais, was optimized and validated. Linearity, precision, recovery, limits of detection and quantification, and selectivity were the parameters used to evaluate performance. The samples were extracted with methanol:water (1: 1 in an ultra-sound bath. The linearity between 0.05 and 20.00 μg/mL (0.5 and 200.0μg of LSD/blotter was observed with satisfactory mean intra and inter assay precision (RSDr = 4.4% and RSD R = 6.4%, respectively and with mean recoveries of 83.4% and 84.9% to the levels of 1.00 and 20.00 μg/mL (10 and 200μg LSD/blotter. The limits of detection and quantification were 0.01 and 0.05 μg/mL, respectively (0.1 and 0.5 μg of LSD/blotter. The samples of blotters (n =22 were analyzed and the mean value of 67.55 μg of LSD/blotter (RSD=27.5% was found. Thus, the method used showed satisfactory analytical performance, and proved suitable as an analytical tool for LSD determination in illicit samples seized by police forces.No presente trabalho, um método utilizando cromatografia líquida de alta eficiência foi otimizado e validado para quantificar o LSD em selos apreendidos em Minas Gerais. A linearidade, precisão, recuperação, limites de detecção e quantificação e seletividade foram os parâmetros de desempenho avaliados. As amostras foram extraídas com metanol: água (1:1 em banho de ultra-som. A linearidade entre 0,05 a 20,00 mg/mL (0,5 a 200 μg LSD/blotter foi observada com precisão média, intra e inter ensaio, satisfatória (RSDr = 4,4% e RSD R = 6,4%, respectivamente e com recuperações médias de 83,4% e 84,9% para os níveis de LSD de 1,00 e 20,00 mg/mL (10 e 200 μg LSD/selo. Os limites de detecção e quantificação encontrados foram de 0,01 e 0,05 mg/mL, respectivamente (0,1 e 0,5 μg LSD/selo. As amostras de selos (n = 22 foram analisadas e o valor médio encontrado foi de 67

  15. Determination of La and Nd by thermal ionization mass spectrometry (TIMS) pre-separated by high performance liquid chromatography (HPLC)

    International Nuclear Information System (INIS)

    Jaison, P.G.; Raut, N.M.; Parab, A.R.; Khodade, P.S.; Govindan, R.; Aggarwal, S.K.

    2003-01-01

    Determination of La and Nd by TIMS is required for accurate determination of burn-up of nuclear fuels. During their thermal ionization mass spectrometric (TIMS) analysis, 138 Ce and 142 Ce show spectroscopic isobaric interferences at 138 La and 142 Nd, respectively. Hence, it is essential to remove Ce from La and Nd for their accurate isotopic composition determination. Reversed phase high performance liquid chromatography (HPLC) is a promising technique for rapid and effective separation

  16. Quantitative Analysis of Ingenol in Euphorbia species via Validated Isotope Dilution Ultra-high Performance Liquid Chromatography Tandem Mass Spectrometry

    Czech Academy of Sciences Publication Activity Database

    Béres, T.; Dragull, K.; Pospíšil, Jiří; Tarkowská, Danuše; Dančák, M.; Bíba, Ondřej; Tarkowski, P.; Doležal, K.; Strnad, Miroslav

    2018-01-01

    Roč. 29, č. 1 (2018), s. 23-29 ISSN 0958-0344 R&D Projects: GA ČR GA17-14007S; GA MŠk(CZ) LO1204 Institutional support: RVO:61389030 Keywords : Euphorbia genus * ingenol * isotope-dilution method * mass spectrometry * ultra-high performance liquid chromatography Subject RIV: FD - Oncology ; Hematology OBOR OECD: Analytical chemistry Impact factor: 2.292, year: 2016

  17. Enantiomeric high-performance liquid chromatography resolution and absolute configuration of 6β-benzoyloxy-3α-tropanol.

    Science.gov (United States)

    Muñoz, Marcelo A; González, Natalia; Joseph-Nathan, Pedro

    2016-07-01

    The absolute configuration of the naturally occurring isomers of 6β-benzoyloxy-3α-tropanol (1) has been established by the combined use of chiral high-performance liquid chromatography with electronic circular dichroism detection and optical rotation detection. For this purpose (±)-1, prepared in two steps from racemic 6-hydroxytropinone (4), was subjected to chiral high-performance liquid chromatography with electronic circular dichroism and optical rotation detection allowing the online measurement of both chiroptical properties for each enantiomer, which in turn were compared with the corresponding values obtained from density functional theory calculations. In an independent approach, preparative high-performance liquid chromatography separation using an automatic fraction collector, yielded an enantiopure sample of OR (+)-1 whose vibrational circular dichroism spectrum allowed its absolute configuration assignment when the bands in the 1100-950 cm(-1) region were compared with those of the enantiomers of esters derived from 3α,6β-tropanediol. In addition, an enantiomerically enriched sample of 4, instead of OR (±)-4, was used for the same transformation sequence, whose high-performance liquid chromatography follow-up allowed their spectroscopic correlation. All evidences lead to the OR (+)-(1S,3R,5S,6R) and OR (-)-(1R,3S,5R,6S) absolute configurations, from where it follows that samples of 1 isolated from Knightia strobilina and Erythroxylum zambesiacum have the OR (+)-(1S,3R,5S,6R) absolute configuration, while the sample obtained from E. rotundifolium has the OR (-)-(1R,3S,5R,6S) absolute configuration. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Detection of Free Polyamines in Plants Subjected to Abiotic Stresses by High-Performance Liquid Chromatography (HPLC).

    Science.gov (United States)

    Gong, Xiaoqing; Liu, Ji-Hong

    2017-01-01

    High-performance liquid chromatography (HPLC) is a sensitive, rapid, and accurate technique to detect and characterize various metabolites from plants. The metabolites are extracted with different solvents and eluted with appropriate mobile phases in a designed HPLC program. Polyamines are known to accumulate under abiotic stress conditions in various plant species and thought to provide protection against oxidative stress by scavenging reactive oxygen species. Here, we describe a common method to detect the free polyamines in plant tissues both qualitatively and quantitatively.

  19. Protective properties of wine products and the role of high performance liquid chromatography in the study of these properties

    International Nuclear Information System (INIS)

    Ulyanova, E V; Larionov, O G; Revina, A A; Andrievskaya, D V; Urusova, L M; Fenin, A A

    2013-01-01

    Data on the biologically active substances present in wines and wine products, the methods of their determination, and changes under chemical, radiation and other types of action are generalized. The role of high performance liquid chromatography in the studies of the protective properties of wines is demonstrated. Particular attention is devoted to problems of counterfeiting of wine products and the possibility to reveal it by using amperometric determination of the antioxidant activity. The bibliography includes 117 references

  20. Synthesis of monodisperse silica microspheres and modification with diazoresin for mixed-mode ultra high performance liquid chromatography separations.

    Science.gov (United States)

    Cong, Hailin; Yu, Bing; Tian, Chao; Zhang, Shuai; Yuan, Hua

    2017-11-01

    Monodisperse silica particles with average diameters of 1.9-2.9 μm were synthesized by a modified Stöber method, in which tetraethyl orthosilicate was continuously supplied to the reaction mixture containing KCl electrolyte, water, ethanol, and ammonia. The obtained silica particles were modified by self-assembly with positively charged photosensitive diazoresin on the surface. After treatment with ultraviolet light, the ionic bonding between silica and diazoresin was converted into covalent bonding through a unique photochemistry reaction of diazoresin. Depending on the chemical structure of diazoresin and mobile phase composition, the diazoresin-modified silica stationary phase showed different separation mechanisms, including reversed phase and hydrophilic interactions. Therefore, a variety of baseline separation of benzene analogues and organic acids was achieved by using the diazoresin-modified silica particles as packing materials in ultra high performance liquid chromatography. According to the π-π interactional difference between carbon rings of fullerenes and benzene rings of diazoresin, C 60 and C 70 were also well separated by ultra-high performance liquid chromatography. Because it has a small size, the ∼2.5 μm monodisperse diazoresin-modified silica stationary phase shows ultra-high efficiency compared with the commercial C 18 -silica high-performance liquid chromatography stationary phase with average diameters of ∼5 μm. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Screening antiallergic components from Carthamus tinctorius using rat basophilic leukemia 2H3 cell membrane chromatography combined with high-performance liquid chromatography and tandem mass spectrometry.

    Science.gov (United States)

    Han, Shengli; Huang, Jing; Cui, Ronghua; Zhang, Tao

    2015-02-01

    Carthamus tinctorius, used in traditional Chinese medicine, has many pharmacological effects, such as anticoagulant effects, antioxidant effects, antiaging effects, regulation of gene expression, and antitumor effects. However, there is no report on the antiallergic effects of the components in C. tinctorius. In the present study, we investigated the antiallergic components of C. tinctorius and its mechanism of action. A rat basophilic leukemia 2H3/cell membrane chromatography coupled online with high-performance liquid chromatography and tandem mass spectrometry method was developed to screen antiallergic components from C. tinctorius. The screening results showed that Hydroxysafflor yellow A, from C. tinctorius, was the targeted component that retained on the rat basophilic leukemia 2H3/cell membrane chromatography column. We measured the amount of β-hexosaminidase and histamine released in mast cells and the key markers of degranulation. The release assays showed that Hydroxysafflor yellow A could attenuate the immunoglobulin E induced release of allergic cytokines without affecting cell viability from 1.0 to 50.0 μM. In conclusion, the established rat basophilic leukemia 2H3 cell membrane chromatography coupled with online high-performance liquid chromatography and tandem mass spectrometry method successfully screened and identified Hydroxysafflor yellow A from C. tinctorius as a potential antiallergic component. Pharmacological analysis elucidated that Hydroxysafflor yellow A is an effective natural component for inhibiting immunoglobulin E-antigen-mediated degranulation. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Analysis of lignans in Magnoliae Flos by turbulent flow chromatography with online solid-phase extraction and high-performance liquid chromatography with tandem mass spectrometry.

    Science.gov (United States)

    Zhou, Xuan; Chen, Cen; Ye, Xiaolan; Song, Fenyun; Fan, Guorong; Wu, Fuhai

    2016-04-01

    In this study, a method coupling turbulent flow chromatography with online solid-phase extraction and high-performance liquid chromatography with tandem mass spectrometry was developed for analyzing the lignans in Magnoliae Flos. By the online pretreatment of turbulent flow chromatography solid-phase extraction, the impurities removal and analytes concentration were automatically processed, and the lignans were separated rapidly and well. Seven lignans of Magnoliae Flos including epieudesmin, magnolin, 1-irioresinol-B-dimethyl ether, epi-magnolin, fargesin aschantin, and demethoxyaschantin were identified by comparing their retention behavior, UV spectra, and mass spectra with those of reference substances or literature data. The developed method was validated, and the good results showed that the method was not only automatic and rapid, but also accurate and reliable. The turbulent flow chromatography with online solid-phase extraction and high-performance liquid chromatography with tandem mass spectrometry method holds a high potential to become an effective method for the quality control of lignans in Magnoliae Flos and a useful tool for the analysis of other complex mixtures. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. High Performance Liquid Chromatography of Some Analgesic Compounds: An Instrumental Analysis Experiment.

    Science.gov (United States)

    Haddad, Paul; And Others

    1983-01-01

    Background information, procedures, and results are provided for an experiment demonstrating techniques of solvent selection, gradient elution, pH control, and ion-pairing in the analysis of an analgesic mixture using reversed-phase liquid chromatography on an octadecylsilane column. Although developed using sophisticated/expensive equipment, less…

  4. Urinary neopterin quantification by reverse-phase high-performance liquid chromatography with ultraviolet detection

    NARCIS (Netherlands)

    Castro, de M.R.; Marco, Di G.S.; Arita, D.Y.; Teixeira, L.C.; Pereira, A.B.; Casarini, D.E.

    2004-01-01

    Neopterin plays an important role in the malignant disease diagnostics. However, the methods employed in neopterin determination are generally difficult and/or time consuming. The aim of this work was to standardize a practical method to quantify neopterin using high-performance liquid

  5. Molecular differences between deuterated and protonated polystyrenes using reversed-phase high-performance liquid chromatography.

    Science.gov (United States)

    Kayillo, Sindy; Gray, Michael J; Shalliker, R Andrew; Dennis, Gary R

    2005-05-06

    Isotopic substitution is a technique used to highlight particular bonds within a molecule for kinetic, spectroscopic and structure analysis. It is presumed that although some properties such as stretching frequencies will not be the same for substituted analogues, the chemical interactions will not vary appreciably as a function of labelling. Reversed-phase liquid chromatography has been used to demonstrate that there are significant differences between the chromatographic behaviour of a sequence of deuterated and protonated oligomeric polystyrenes. Two-dimensional reversed-phase liquid chromatography was used to show that even the diasteromers of the oligomers (n = 5) have retention mechanisms that are dependent on the subtle changes to the molecular conformation and electronic structure, which are a consequence of deuteration.

  6. Determination of a Jet Fuel Metal Deactivator by High Performance Liquid Chromatography

    Science.gov (United States)

    1983-06-01

    bonded phase chromatography (Reference 2). 73 AFWAL-TR-82-2128 Bonded phase packings offer distinct advantages over other packings: a. Irreversible...were then oven dried and placed in a dessicator for cooling and storage until use. The bottles were subsequently silanized with "Glas-TREET" ( Alltech ... advantages of a loop injector are: (1) The volume injected is far more repeatable since a fixed volume loop has a constant volume and is flushed with a

  7. High-performance liquid ion-pair chromatography in inorganic analysis

    International Nuclear Information System (INIS)

    Alimarin, I.P.; Basova, E.M.; Bol'shova, T.A.; Ivanov, V.M.

    1990-01-01

    In literature review for the recent 15 years theoretical foundations, regularities and mechanisms of ionized compound retention in reverse-phase ion-pair chromatography are considered, possibilities and prospects of its application in inorganic analysis being demonstrated. Analytic characteristics of the methods for the determination of inorganic anions (I - , IO 3 - , MoO 4 2- , etc.), as well as metals (Zr, Hf, V, Nb, Mo, W, Ru, lanthanides, etc.) in the form of chelates, are given

  8. Determination of gangliosides as 2,4-dinitrophenylhydrazides by high-performance liquid chromatography.

    OpenAIRE

    Miyazaki, K; Okamura, N; Kishimoto, Y; Lee, Y C

    1986-01-01

    A specific, sensitive and easily performed method for the determination of gangliosides in tissue was developed. After removal of water-soluble compounds, total lipids were extracted from tissue and then treated with 2,4-dinitrophenylhydrazine hydrochloride and dicyclohexylcarbodi-imide in dimethylformamide at 0 degrees C to form ganglioside hydrazides. After removal of excess reagents by column chromatography on silicic acid, the ganglioside 2,4-dinitrophenylhydrazides were eluted from the c...

  9. Optimization of the combination micro-high-performance liquid-chromatography/mass spectrometry

    International Nuclear Information System (INIS)

    Haider, K.

    1997-03-01

    The coupling of liquid chromatography and mass spectrometry is still growing in significance. In this thesis, a particle beam interface has been investigated for combining ion chromatography with mass spectrometric detection. To introduce the eluent directly (without membrane suppressor) into the spectrometer, only methods with low flow rates like microcolumn chromatography can be used. For the preparation of the columns, reversed-phase and silica-based anion exchange materials were packed into PEEK, steel and fused-silica capillaries with i.d. from 130 to 1000 μm using different methods. The performance of the particle beam interface (modified with a new miniaturized aerosol generator) and the mass spectrometric detection has been studied for a series of inorganic anions as well as aminopolycarboxylic acids and the metal-EDTA complexes. Detection limits between 10 and 100 ng injected could be achieved in the multiple ion detection mode of the mass spectrometer for the investigated solutes. A second type of interface, the direct liquid introduction (DLI) has been used to analyze the priority pollutant phenols. This interface is based on a modified GC-interface into the MS. Separation columns used so far include packed fused-silica capillaries with inner diameter of 75 μm and polystyrene-divinylbenzene (functionalized with tert. butyl groups) as stationary phase. Aspects of instrumentation and effects of chemical ionization in the direct liquid introduction mode are discussed. (author)

  10. Identification, characterization, and high-performance liquid chromatography quantification of process-related impurities in vonoprazan fumarate.

    Science.gov (United States)

    Liu, Lei; Cao, Na; Ma, Xingling; Xiong, Kaihe; Sun, Lili; Zou, Qiaogen

    2016-04-01

    High-performance liquid chromatography analysis of vonoprazan fumarate, a novel proton pump inhibitor drug revealed six impurities. These were identified by liquid chromatography with mass spectrometry. Further, the structures of the impurities were confirmed by synthesis followed by characterization by mass spectrometry, NMR spectroscopy, and infrared spectroscopy. On the basis of these data and knowledge of the synthetic scheme of vonoprazan fumarate, the previously unknown impurity was identified as 1-[5-(2-fluorophenyl)-1-(pyridin-3-ylsulfonyl)-1H-pyrrol-3-yl]-N-methyldimethylamine, which is a new compound. The possible mechanisms by which these impurities were formed were also discussed. A high-performance liquid chromatography method was optimized in order to separate, selectively detect, and quantify all process-related impurities of vonoprazan fumarate. The presented method has been validated in terms of linearity, limits of detection, and quantification, and response factors and, therefore, is highly suitable for routine analysis of vonoprazan fumarate related substances as well as stability studies. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Ultra‐high performance supercritical fluid chromatography of lignin‐derived phenols from alkaline cupric oxide oxidation

    Science.gov (United States)

    Sun, Mingzhe; Lidén, Gunnar; Sandahl, Margareta

    2016-01-01

    Traditional chromatographic methods for the analysis of lignin‐derived phenolic compounds in environmental samples are generally time consuming. In this work, an ultra‐high performance supercritical fluid chromatography method with a diode array detector for the analysis of major lignin‐derived phenolic compounds produced by alkaline cupric oxide oxidation was developed. In an analysis of a collection of 11 representative monomeric lignin phenolic compounds, all compounds were clearly separated within 6 min with excellent peak shapes, with a limit of detection of 0.5–2.5 μM, a limit of quantification of 2.5–5.0 μM, and a dynamic range of 5.0–2.0 mM (R 2 > 0.997). The new ultra‐high performance supercritical fluid chromatography method was also applied for the qualitative and quantitative analysis of lignin‐derived phenolic compounds obtained upon alkaline cupric oxide oxidation of a commercial humic acid. Ten out of the previous eleven model compounds could be quantified in the oxidized humic acid sample. The high separation power and short analysis time obtained demonstrate for the first time that supercritical fluid chromatography is a fast and reliable technique for the analysis of lignin‐derived phenols in complex environmental samples. PMID:27452148

  12. Ultra-high performance supercritical fluid chromatography of lignin-derived phenols from alkaline cupric oxide oxidation.

    Science.gov (United States)

    Sun, Mingzhe; Lidén, Gunnar; Sandahl, Margareta; Turner, Charlotta

    2016-08-01

    Traditional chromatographic methods for the analysis of lignin-derived phenolic compounds in environmental samples are generally time consuming. In this work, an ultra-high performance supercritical fluid chromatography method with a diode array detector for the analysis of major lignin-derived phenolic compounds produced by alkaline cupric oxide oxidation was developed. In an analysis of a collection of 11 representative monomeric lignin phenolic compounds, all compounds were clearly separated within 6 min with excellent peak shapes, with a limit of detection of 0.5-2.5 μM, a limit of quantification of 2.5-5.0 μM, and a dynamic range of 5.0-2.0 mM (R(2) > 0.997). The new ultra-high performance supercritical fluid chromatography method was also applied for the qualitative and quantitative analysis of lignin-derived phenolic compounds obtained upon alkaline cupric oxide oxidation of a commercial humic acid. Ten out of the previous eleven model compounds could be quantified in the oxidized humic acid sample. The high separation power and short analysis time obtained demonstrate for the first time that supercritical fluid chromatography is a fast and reliable technique for the analysis of lignin-derived phenols in complex environmental samples. © 2016 The Authors, Journal of Separation Science Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Determination of pentachlorophenol in water and aquifer sediments by high-performance liquid chromatography

    Science.gov (United States)

    Goerlitz, D.F.

    1981-01-01

    Methods for the determination of pentachlorophenol (PCP) in water and aquifer sediments are presented. Reverse-phase high-performance liquid chromotography employing ion suppression and gradient elution is used. PCP can be determined directly in water at a lower limit of detection Of 0.2 micrograms per liter. For extracts of sediment, PCP can be determined to a lower limit of 1.0 micrograms per kilogram.

  14. Gas chromatography-mass spectrometry and high-performance liquid chromatography-diode array detection for dating of paper ink.

    Science.gov (United States)

    Díaz-Santana, Oscar; Vega-Moreno, Daura; Conde-Hardisson, Francisco

    2017-09-15

    An extraction and determination method is shown for the analysis of dyes and solvents present in two types of ballpoint pen inks that are deposited onto paper. Ink extracts are analysed using a combination of gas chromatography with mass spectrometry (GC-MS), and high-pressure liquid chromatography with photodiode array detection (HPLC-DAD), within a single sample extraction procedure. Seventeen solvents and thirteen dyes contained in two Montblanc ® inks (black and blue) were monitored for 45 months at monthly intervals, in order to determine variations in the concentrations of the compounds over time. We also studied the relative variations between different compounds and the generation of degradation products such as phenol. The concentration data obtained from these compounds during their exposure have been analysed and a multiple regression model is developed for each ink type that allows an estimate of the exposure time of the ink on paper with a maximum error of between 4 and 7 months. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Determination of limonin and nomilin contents in different citrus cultivars using high performance liquid chromatography

    International Nuclear Information System (INIS)

    Bilal, H.; Hassan, S.; Sahar, S.; Akram, W.; Sahar, S.

    2013-01-01

    High perlorrnance liquid chromatography (HPLC) analysis was done to quantify the amount of limonoids (nomilin and nomilin) in seven selected citrus cultivars. According to the HPLC analysis red blood orange (Citrus sinensis var red blood orange) had maximum amount of limonin (479.77 ug/rnl.), while rough lemon (Citrus jambhiri) had no limonin content. in case of nomonin, rough lemon (Citrus jambhir) had maximum amount of nomilin (54.23 micro g/ML)) while succari (citrus sinensis var succari) had very low amount of nomilin (0.37 micro g/Ml). (author)

  16. Skeletal keratan sulfate chain molecular weight calibration by high-performance gel-permeation chromatography

    International Nuclear Information System (INIS)

    Dickenson, J.M.; Morris, H.G.; Nieduszynski, I.A.; Huckerby, T.N.

    1990-01-01

    A method has been developed for the molecular sizing of skeletal keratan sulfate chains using an HPLC gel-permeation chromatography system. Keratan sulfate chains and keratanase-derived oligosaccharides were prepared from the nucleus pulposus of bovine intervertebral disc (6-year-old animals). A Bio-Gel TSK 30 XL column eluted in 0.2 M NaCl and at 30 degrees C was calibrated with keratan sulfate oligosaccharides of known size as well as 3H-end-labeled keratan sulfate chains to yield the relationship

  17. Validation of a technique by high-performance liquid chromatography for the determination of total isoflavones

    Directory of Open Access Journals (Sweden)

    Pilar A. Soledispa Cañarte

    2017-04-01

    Full Text Available Context: Isoflavones may act as selective regulators in the prevention of various diseases. The most important source of isoflavones is the soy, from which different phytotherapeutics are elaborated of use in Ecuadorian population. However, its concentration varies depending on several factors, therefore quality assessment need to be carried out through out several analytical methods. Aims: To validate an analytical method by high precision liquid chromatography (HPLC to quantify total isoflavones in herbal medicine. Methods: To quantify isoflavones, it was used a brand liquid chromatography with UV/VIS detector at 260 nm, C-18 column using isocratic method. The mobile phase was composed of 2% acetic acid: acetonitrile (75:25. The quantification was performed against reference standard. The parameters for the validation followed the established in the USP 33. Results: The chromatogram presented six peaks with elution between 1.557 and 18.913 min. The linearity of the system and the method got r2 equal to 0.98 and 0.99 respectively. The coefficients of variation 1.5% in the study of repetitiveness and 2% in intermediate precision. The accuracy of the adjusted lineal model exhibited r=0.95 and intercept reliable interval (-0.921; 1.743. Conclusions: The validated method was specific, accurate, precise and linear. It can be used for quality control and stability studies of isoflavones present in herbal medicine.

  18. Comprehensive sample analysis using high performance liquid chromatography with multi-detection

    International Nuclear Information System (INIS)

    Pravadali, Sercan; Bassanese, Danielle N.; Conlan, Xavier A.; Francis, Paul S.; Smith, Zoe M.; Terry, Jessica M.; Shalliker, R. Andrew

    2013-01-01

    Graphical abstract: -- Highlights: •Detection selectivity was assessed with 6 detection modes. •Natural samples show great diversity in detection selectivity. •Complex samples require evaluation using a multifaceted approach to detection. •23/30 known compounds (detected by MS) detected by chemiluminescence, DPPH and UV. -- Abstract: Herein we assess the separation space offered by a liquid chromatography system with an optimised uni-dimensional separation for the determination of the key chemical entities in the highly complex matrix of a tobacco leaf extract. Multiple modes of detection, including UV–visible absorbance, chemiluminescence (acidic potassium permanganate, manganese(IV), and tris(2,2′-bipyridine)ruthenium(III)), mass spectrometry and DPPH radical scavenging were used in an attempt to systematically reduce the data complexity of the sample whilst obtaining a greater degree of molecule-specific information. A large amount of chemical data was obtained, but several limitations in the ability to assign detector responses to particular compounds, even with the aid of complementary detection systems, were observed. Thirty-three compounds were detected via MS on the tobacco extract and 12 out of 32 compounds gave a peak height ratio (PHR) greater than 0.33 on one or more detectors. This paper serves as a case study of these limitations, illustrating why multidimensional chromatography is an important consideration when developing a comprehensive chemical detection system

  19. Comprehensive sample analysis using high performance liquid chromatography with multi-detection.

    Science.gov (United States)

    Pravadali, Sercan; Bassanese, Danielle N; Conlan, Xavier A; Francis, Paul S; Smith, Zoe M; Terry, Jessica M; Shalliker, R Andrew

    2013-11-25

    Herein we assess the separation space offered by a liquid chromatography system with an optimised uni-dimensional separation for the determination of the key chemical entities in the highly complex matrix of a tobacco leaf extract. Multiple modes of detection, including UV-visible absorbance, chemiluminescence (acidic potassium permanganate, manganese(IV), and tris(2,2'-bipyridine)ruthenium(III)), mass spectrometry and DPPH radical scavenging were used in an attempt to systematically reduce the data complexity of the sample whilst obtaining a greater degree of molecule-specific information. A large amount of chemical data was obtained, but several limitations in the ability to assign detector responses to particular compounds, even with the aid of complementary detection systems, were observed. Thirty-three compounds were detected via MS on the tobacco extract and 12 out of 32 compounds gave a peak height ratio (PHR) greater than 0.33 on one or more detectors. This paper serves as a case study of these limitations, illustrating why multidimensional chromatography is an important consideration when developing a comprehensive chemical detection system. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Comprehensive sample analysis using high performance liquid chromatography with multi-detection

    Energy Technology Data Exchange (ETDEWEB)

    Pravadali, Sercan [Australian Centre for Research on Separation Sciences (ACROSS), School of Science and Health, University of Western Sydney (Parramatta), NSW 1797 (Australia); Bassanese, Danielle N.; Conlan, Xavier A.; Francis, Paul S.; Smith, Zoe M.; Terry, Jessica M. [Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences, Deakin University, Victoria 3216 (Australia); Shalliker, R. Andrew, E-mail: R.Shalliker@uws.edu.au [Australian Centre for Research on Separation Sciences (ACROSS), School of Science and Health, University of Western Sydney (Parramatta), NSW 1797 (Australia)

    2013-11-25

    Graphical abstract: -- Highlights: •Detection selectivity was assessed with 6 detection modes. •Natural samples show great diversity in detection selectivity. •Complex samples require evaluation using a multifaceted approach to detection. •23/30 known compounds (detected by MS) detected by chemiluminescence, DPPH and UV. -- Abstract: Herein we assess the separation space offered by a liquid chromatography system with an optimised uni-dimensional separation for the determination of the key chemical entities in the highly complex matrix of a tobacco leaf extract. Multiple modes of detection, including UV–visible absorbance, chemiluminescence (acidic potassium permanganate, manganese(IV), and tris(2,2′-bipyridine)ruthenium(III)), mass spectrometry and DPPH radical scavenging were used in an attempt to systematically reduce the data complexity of the sample whilst obtaining a greater degree of molecule-specific information. A large amount of chemical data was obtained, but several limitations in the ability to assign detector responses to particular compounds, even with the aid of complementary detection systems, were observed. Thirty-three compounds were detected via MS on the tobacco extract and 12 out of 32 compounds gave a peak height ratio (PHR) greater than 0.33 on one or more detectors. This paper serves as a case study of these limitations, illustrating why multidimensional chromatography is an important consideration when developing a comprehensive chemical detection system.

  1. Ion exchange of alkaline metals on the thin-layer zinc ferrocyanide

    International Nuclear Information System (INIS)

    Betenekov, N.D.; Buklanov, G.V.; Ipatova, E.G.; Korotkin, Yu.S.

    1991-01-01

    Basic regularities of interphase distribution in the system of thin-layer sorbent on the basis of mixed zinc ferrocyanide (FZ)-alkaline metal solution (Na, K, Rb, Cs, Fr) in the column chromatography made are studied. It is established that interphase distribution of microgram amounts of alkaline metals in the systems thin-layer FZ-NH 4 NO 3 electrolyte solutions is of ion-exchange character and subjected to of law effective mass. It is shown that FZ thin-layer material is applicable for effective chromatographic separation of alkaline metal trace amounts. An approach to the choice of a conditions of separate elution of Na, K, Rb, Cs, Fr in the column chromatography mode

  2. Analysis by high-performance liquid chromatography of teucrin A in beverages flavoured with an extract of Teucrium chamaedrys L.

    Science.gov (United States)

    Bosisio, E; Giavarini, F; Dell'Agli, M; Galli, G; Galli, C L

    2004-05-01

    Due to its liver toxicity, the medicinal use of germander (Teucrium chamaedrys L.) was banned in some countries. Nevertheless, alcoholic extracts are still permitted as flavour ingredients since they are fundamental in providing a bitter aromatic taste. Teucrin A represents the substance of major concern regarding the potential toxicity of germander. Hence, teucrin A represents the best analytical and toxicological marker of alcoholic extracts of T. chamaedrys. A sensitive high-performance liquid chromatography method to detect teucrin A in beverages is reported. Teucrin A was prepared by isolation from the plant extract using column chromatography and crystallization. The identity and purity (99%) were established by melting point, nuclear magnetic resonance and liquid chromatography-mass spectrometry. The high-performance liquid chromatography procedure was validated and its intra- and interday performance was established (relative standard deviation beverages not containing T. chamaedrys spiked with a range of concentrations of teucrin A. The limit of detection was 0.1 ppm and the limit of quantification was 0.3 ppm. Teucrin A accounted for about 70% of the neo-clerodane diterpenoids found in the total extract of a specimen of T. chamaedrys. The content (+/- standard deviation) in 18 batches of different geographical origin was 2338 +/- 740 ppm, per cent coefficient of variation = 32, minimum-maximum = 999 - 3445 ppm. The mean level of teucrin A in 10 bottles of the same brand was 6.1 +/- 0.8 ppm, per cent coefficient of variation = 12. In 10 different brands found on the Italian market, the content of teucrin A ranged from not detectable to 10 ppm.

  3. Investigation into the temporal stability of aqueous standard solutions of psilocin and psilocybin using high performance liquid chromatography.

    Science.gov (United States)

    Anastos, N; Barnett, N W; Pfeffer, F M; Lewis, S W

    2006-01-01

    This paper reports an investigation into the temporal stability of aqueous solutions of psilocin and psilocybin reference drug standards over a period of fourteen days. This study was performed using high performance liquid chromatography utilising a (95:5% v/v) methanol: 10 mM ammonium formate, pH 3.5 mobile phase and absorption detection at 269 nm. It was found that the exclusion of light significantly prolonged the useful life of standards, with aqueous solutions of both psilocin and psilocybin being stable over a period of seven days.

  4. L-ascorbic acid losses in Kenyan vegetables during cooking as determined by high performance liquid chromatography

    OpenAIRE

    N.M.N. Wekesa; S.C. Chhabra; H.M. Thairu

    2001-01-01

    The loss of L-ascorbic acid (L-AA) in 14 different cooked local vegetables found in Nairobi markets was determined by high performance liquid chromatography. The effect of quantity of water on the loss of L-AA during cooking was studied with cowpea leaves. It was found that more L-AA was lost when larger amount of water was used than when smaller amount was used. The effect of the sharpness of the knife on the loss of L-AA was studied with spinach. It was found that more loss of L-AA occurred...

  5. Analysis of human milk oligosaccharides using high-performance anion-exchange chromatography with pulsed amperometric detection

    DEFF Research Database (Denmark)

    Lie, Aleksander; Pedersen, Lars Haastrup

    ) and lacto-N-neotetraose (Galβ1-4GlcNAcβ1-3Galβ1-4Glc), among others. High-performance anion-exchange chromatography (HPAE) with pulsed amperometric detection (PAD) is an analysis method highly suited for carbohydrates. HPAE with alkaline eluents results in retention of neutral carbohydrates depending...... on the number of charged group in the molecule, pH and concentration of competing anions, while the PAD has sensitivity for carbohydrates in the pmol-range (Lee 1990). As a basis for the development and optimisation of HPAE elution methods, the parameter space was investigated in terms of eluent concentrations...

  6. Separation of human milk oligosaccharides using high-performance anion-exchange chromatography with pulsed amperometric detection

    DEFF Research Database (Denmark)

    Lie, Aleksander; Pedersen, Lars Haastrup

    individual mothers is considerable, ranging from as few as 23 and up to 130 different oligosaccharides. HMOs are known as beneficial for infant health and development, and have received increasing attention in recent years (Bode & Jantscher-Krenn 2012). High-performance anion-exchange chromatography (HPAE......) with pulsed amperometric detection (PAD) is an analysis method highly suited for carbohydrates. HPAE with alkaline eluents results in retention of neutral carbohydrates depending on the number of charged groups in the molecule, pH and concentration of competing anions, while PAD has sensitivity...

  7. Determination of 2-naphthylamine in urine by a novel reversed-phase high-performance liquid chromatography method

    DEFF Research Database (Denmark)

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

    1992-01-01

    A high-performance liquid chromatographic method for the determination of 2-naphthylamine in urine using fluorescence detection was developed. The method validation analysis showed the method to be in analytical control, i.e. the distribution of the difference between the observed and true values...... of the method evaluation samples did not deviate significantly from the normal distribution. The recovery of the method was 85%. The entire run time of chromatography was 10 min using isocratic elution (acetonitrile-water, 35:65), and the retention time for 2-naphthylamine was 5.8 min. The relative short time...

  8. High-Performance Liquid Chromatography (HPLC)-Based Detection and Quantitation of Cellular c-di-GMP.

    Science.gov (United States)

    Petrova, Olga E; Sauer, Karin

    2017-01-01

    The modulation of c-di-GMP levels plays a vital role in the regulation of various processes in a wide array of bacterial species. Thus, investigation of c-di-GMP regulation requires reliable methods for the assessment of c-di-GMP levels and turnover. Reversed-phase high-performance liquid chromatography (RP-HPLC) analysis has become a commonly used approach to accomplish these goals. The following describes the extraction and HPLC-based detection and quantification of c-di-GMP from Pseudomonas aeruginosa samples, a procedure that is amenable to modifications for the analysis of c-di-GMP in other bacterial species.

  9. High-performance liquid chromatography analysis methods developed for quantifying enzymatic esterification of flavonoids in ionic liquids

    DEFF Research Database (Denmark)

    Lue, Bena-Marie; Guo, Zheng; Xu, X.B.

    2008-01-01

    Methods using reversed-phase high-performance liquid chromatography (RP-HPLC) with ELSD were investigated to quantify enzymatic reactions of flavonoids with fatty acids in the presence of diverse room temperature ionic liquids (RTILs). A buffered salt (preferably triethylamine-acetate) was found...... essential for separation of flavonoids from strongly polar RTILs, whereby RTILs were generally visible as two major peaks identified based on an ion-pairing/exchanging hypothesis. C8 and C12 stationary phases were optimal while mobile phase pH (3-7) had only a minor influence on separation. The method...

  10. Separation of deuteriated isotopomers of dopamine by ion-pair reversed-phase high-performance liquid chromatography

    International Nuclear Information System (INIS)

    Masters, C.F.; Markey, S.P.; Mefford, I.N.; Duncan, M.W.

    1988-01-01

    The ion-pair reversed-phase separation of dopamine and deuterium-substituted dopamine isotopomers is described. Chromatographic parameters and deuterium isotope effects governing the resolution are examined and compared to the factors regulating the resolution are examined and compared to the factors regulating the resolution of the chemically distinct entities dopamine, norepinephrine, and epinephrine. The potential utility of the [ 2 H 7 ]dopamine, isotopomer as an internal standard for the high-performance liquid chromatography analysis of dopamine is demonstrated by using aluminum oxide extraction prior to chromatographic separation

  11. [Determination by high performance chromatography, steroid saponins in a biologically active food supplements containing the extract of Tribulus terrestris].

    Science.gov (United States)

    Kozlova, O I; Perederiaev, O I; Ramenskaia, G V

    2011-01-01

    Steroidal saponins are bioactive substances of Tribulus terrestris and can be used to assess the quality of raw materials and processed products from them. For this purpose has been developed the method of qualitative and quantitative determination of steroidal saponins by high performance liquid chromatography with spectrophotometric and mass-selective detection and optimal conditions of sample preparation (70% methanol extraction with sonication and heating); also has been studied steroidal saponins composition of Tribulus terrestris (protodioscin, tribulosaponin B, metilprotodiostsin, terrestrozin H, prototribestin, gracillin and others were found).

  12. Analysis of proinsulin and its conversion products by reversed-phase high-performance liquid chromatography

    DEFF Research Database (Denmark)

    Linde, S; Welinder, B S; Nielsen, Jens Høiriis

    1993-01-01

    . Most mammals produce a single insulin, but in rodents two non-allelic insulin genes are expressed. There is an inverse ratio between the two insulins in rats and mice, the reason for this being unknown. It has been suggested that differences in transcription, translation (biosynthesis) and...... PIM (intact proinsulin or its intermediates) has been incompletely determined. Studies of the biosynthesis of proinsulins and their conversion with the purpose of revealing some of these points depend on accessible reversed-phase high-performance liquid chromatographic (RP-HPLC) analyses capable...

  13. Phytochemical analysis of Hibiscus caesius using high performance liquid chromatography coupled with mass spectrometry.

    Science.gov (United States)

    Ain, Quratul; Naveed, Muhammad Na; Mumtaz, Abdul Samad; Farman, Muhammad; Ahmed, Iftikhar; Khalid, Nauman

    2015-09-01

    Various species in genus Hibiscus are traditionally known for their therapeutic attributes. The present study focused on the phytochemical analysis of a rather unexplored species Hibiscus caesius (H. caesius), using high-pressure liquid chromatography coupled with mass spectrometry (HPLC-MS). The analysis revealed five major compounds in the aqueous extract, viz. vanillic acid, protocatechoic acid, quercetin, quercetin glucoside and apigenin, being reported for the first time in H. caesius. Literature suggests that these compounds have important pharmacological traits such as anti-cancer, anti-inflammatory, anti-bacterial and hepatoprotective etc. however, this requires further pharmacological investigations at in vitro and in vivo scale. The above study concluded the medicinal potential of H. caesius.

  14. Melatonin in edible plants identified by radioimmunoassay and by high performance liquid chromatography-mass spectrometry

    International Nuclear Information System (INIS)

    Dubbels, R.; Klenke, E.; Schnakenberg, E.; Ehlers, C.; Schloot, W.; Reiter, R.J.; Goebel, A.; Schiware, H.W.

    1995-01-01

    Melatonin, the chief hormone of the pineal gland in vertebrates, is widely distributed in the animal kingdom. Among many functions, melatonin synchronizes circadian and circannual rhythms, stimulates immune function, may increase life span, inhibits growth of cancer cells in vitro and cancer progression and promotion in vivo, and was recently shown to be a potent hydroxyl radical scavenger and antioxidant. Hydroxyl radicals are highly toxic by-products of oxygen metabolism that damage cellular DNA and other macromolecules. Herein we report that melatonin, in varying concentrations, is also found in a variety of plants. Melatonin concentrations, measured in nine different plants by radioimmunoassay, ranged from 0 to 862 pg melatonin/mg protein. The presence of melatonin was verified by gas chromatography/mass spectrometry. Our findings suggest that the consumption of plant materials that contain high levels of melatonin could alter blood melatonin levels of the indole as well as provide protection of macromolecules against oxidative damage. (au) 30 refs

  15. Determination of urinary 2- and 3-dechloroethylated metabolites of ifosfamide by high-performance liquid chromatography.

    Science.gov (United States)

    Goren, M P

    1991-10-04

    In vivo oxidation of chloroethyl side-chains on ifosfamide produces the toxin chloroacetaldehyde. Production of this labile metabolite can be indirectly quantitated by monitoring the excretion of the residual 2- and 3-dechloroethylated ifosfamide. Urinary ifosfamide and the two dechloroethylated metabolites were extracted into chloroform from alkalinized salt-saturated urine, followed by high-performance liquid chromatographic separation using an acetonitrile gradient on a reversed-phase column and ultraviolet detection at 190 nm. In five patients given 1.6 g/m2 ifosfamide, 11-30% of the dose was excreted over 24 h as unchanged drug, 11-21% as 3-dechloroethylated and 3-10% as 2-dechloroethylated ifosfamide.

  16. Determination of tylosin residues in pig tissues using high-performance liquid chromatography.

    Science.gov (United States)

    De Liguoro, M; Anfossi, P; Angeletti, R; Montesissa, C

    1998-06-01

    In accordance with the maximum residue limit of 100 micrograms kg-1 established by EU legislation, a simple and sensitive high-performance liquid chromatographic (HPLC) method was developed for the measurement of tylosin residues in pig tissues (fat, kidney, liver and muscle). Tylosin, a macrolide antibiotic, is extracted with water-methanol and cleaned-up by solid-phase extraction (SPE) on cation-exchange cartridges using methanol elution. Tylosin was determined by reversed-phase HPLC with UV detection at 280 nm and the mean recovery from pig tissues fortified in the range 50-200 micrograms kg-1 was 70-85%, with intra- and inter-day RSDs in the ranges 3.4-9.1 and 3.9-10.1% respectively.

  17. High-performance liquid chromatography with fluorescence detection for the rapid analysis of pheophytins and pyropheophytins in virgin olive oil.

    Science.gov (United States)

    Li, Xueqi; Woodman, Michael; Wang, Selina C

    2015-08-01

    Pheophytins and pyropheophytin are degradation products of chlorophyll pigments, and their ratios can be used as a sensitive indicator of stress during the manufacturing and storage of olive oil. They increase over time depending on the storage condition and if the oil is exposed to heat treatments during the refining process. The traditional analysis method includes solvent- and time-consuming steps of solid-phase extraction followed by analysis by high-performance liquid chromatography with ultraviolet detection. We developed an improved dilute/fluorescence method where multi-step sample preparation was replaced by a simple isopropanol dilution before the high-performance liquid chromatography injection. A quaternary solvent gradient method was used to include a fourth strong solvent wash on a quaternary gradient pump, which avoided the need to premix any solvents and greatly reduced the oil residues on the column from previous analysis. This new method not only reduces analysis cost and time but shows reliability, repeatability, and improved sensitivity, especially important for low-level samples. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Strategy for reduced calibration sets to develop quantitative structure-retention relationships in high-performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Andries, Jan P.M. [University of Professional Education, Department of Life Sciences, P.O. Box 90116, 4800 RA Breda (Netherlands); Claessens, Henk A. [University of Professional Education, Department of Life Sciences, P.O. Box 90116, 4800 RA Breda (Netherlands); Eindhoven University of Technology, Department of Chemical Engineering and Chemistry, Laboratory of Polymer Chemistry, P.O. Box 513 (Helix, STW 1.35), 5600 MB Eindhoven (Netherlands); Heyden, Yvan Vander [Department of Analytical Chemistry and Pharmaceutical Technology, Vrije Universiteit Brussel-VUB, Laarbeeklaan 103, B-1090 Brussels (Belgium); Buydens, Lutgarde M.C., E-mail: L.Buydens@science.ru.nl [Institute for Molecules and Materials, Radboud University Nijmegen, Toernooiveld 1, 6525 ED Nijmegen (Netherlands)

    2009-10-12

    In high-performance liquid chromatography, quantitative structure-retention relationships (QSRRs) are applied to model the relation between chromatographic retention and quantities derived from molecular structure of analytes. Classically a substantial number of test analytes is used to build QSRR models. This makes their application laborious and time consuming. In this work a strategy is presented to build QSRR models based on selected reduced calibration sets. The analytes in the reduced calibration sets are selected from larger sets of analytes by applying the algorithm of Kennard and Stone on the molecular descriptors used in the QSRR concerned. The strategy was applied on three QSRR models of different complexity, relating logk{sub w} or log k with either: (i) log P, the n-octanol-water partition coefficient, (ii) calculated quantum chemical indices (QCI), or (iii) descriptors from the linear solvation energy relationship (LSER). Models were developed and validated for 76 reversed-phase high-performance liquid chromatography systems. From the results we can conclude that it is possible to develop log P models suitable for the future prediction of retentions with as few as seven analytes. For the QCI and LSER models we derived the rule that three selected analytes per descriptor are sufficient. Both the dependent variable space, formed by the retention values, and the independent variable space, formed by the descriptors, are covered well by the reduced calibration sets. Finally guidelines to construct small calibration sets are formulated.

  19. Comprehensive analysis of chemical constituents in Xingxiong injection by high performance liquid chromatography coupled with mass spectrometry.

    Science.gov (United States)

    Guo, Long; Dou, Li-Li; Duan, Li; Liu, Ke; Bi, Zhi-Ming; Li, Ping; Liu, E-Hu

    2015-09-01

    Xingxiong injection (XXI) is a widely used Chinese herbal formula prepared by the folium ginkgo extract and ligustrazine for the treatment of cardiovascular and cerebrovascular diseases. Compared with the pharmacological studies, chemical analysis and quality control studies on this formula are relatively limited. In the present study, a high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-QTOF MS) method was applied to comprehensive analysis of constituents in XXI. According to the fragmentation rules and previous reports, thirty ginkgo flavonoids, four ginkgo terpene lactones, and one alkaloid were identified. A high performance liquid chromatography coupled with triple quadrupole mass spectrometry (HPLC-QQQ MS) method was then applied to quantify ten major constituents in XXI. The method validation results indicated that the developed method had desirable specificity, linearity, precision and accuracy. The total contents of ginkgo flavonoids were about 22.05-25.51 μg·mL(-1) and the ginkgo terpene lactones amounts were about 4.41-8.70 μg·mL(-1) in six batches of XXI samples, respectively. Furthermore, cosine ratio algorithm and distance measurements were employed to evaluate the similarity of XXI samples, and the results demonstrated a high-quality consistency. This work could provide comprehensive information on the quality control of Xingxiong injection, which be helpful in the establishment of a rational quality control standard. Copyright © 2015 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.

  20. Proteomic analysis of Taenia ovis metacestodes by high performance liquid chromatography-coupled tandem mass spectrometry.

    Science.gov (United States)

    Zheng, Yadong

    2017-03-15

    Taenia ovis metacestodes reside in the muscle of sheep and goats, and may cause great economic loss due to condemnation of carcasses if not effectively controlled. Although advances have been made in the control of T. ovis infection, our knowledge of T. ovis biology is limited. Herein the protein profiling of T. ovis metacestodes was determined by liquid chromatography-linked tandem mass spectrometry. A total of 966 proteins were identified and 25.1% (188/748) were annotated to be associated with metabolic pathways. Consistently, GO analysis returned a metabolic process (16.27%) as one of two main biological process terms. Moreover, it was found that 24 proteins, including very low-density lipoprotein receptor, enolase, paramyosin and endophilin B1, were abundant in T. ovis metacestodes. These proteins may be associated with motility, metabolism, signaling, stress, drug resistance and immune responses. Furthermore, comparative analysis of 5 cestodes revealed the presence of Taenia-specific enolases. These data provide clues for better understanding of T. ovis biology, which is informative for effective control of infection. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Hazards of Secondary Bromadiolone Intoxications Evaluated using High-performance Liquid Chromatography with Electrochemical Detection

    Directory of Open Access Journals (Sweden)

    René Kizek

    2007-07-01

    Full Text Available This study reported on the possibility of intoxications of non-target wild animalsassociated with use of bromadiolone as the active component of rodenticides withanticoagulation effects. A laboratory test was done with earthworms were exposed tobromadiolone-containing granules under the conditions specified in the modified OECD207 guideline. No mortality of earthworms was observed during the fourteen days longexposure. When the earthworms from the above test became a part of the diet of commonvoles in the following experiment, no mortality of consumers was observed too. However,electrochemical analysis revealed higher levels of bromadiolone in tissues fromearthworms as well as common voles compared to control animals. There were determinedcomparable levels of bromadiolone in the liver tissue of common voles after primary(2.34±0.10 μg/g and secondary (2.20±0.53 μg/g intoxication. Therefore, the risk ofsecondary intoxication of small mammalian species feeding on bromadiolone-containing earthworms is the same as of primary intoxication through baited granules. Bromadiolone bio-accumulation in the food chain was monitored using the newly developed analytical procedure based on the use of a liquid chromatography coupled with electrochemical detector (HPLC-ED. The HPLC-ED method allowed to determine the levels of bromadiolone in biological samples and is therefore suitable for examining the environmental hazards of this substance.

  2. Determination of drug lipophilicity by phosphatidylcholine-modified microemulsion high-performance liquid chromatography.

    Science.gov (United States)

    Xuan, Xueyi; Xu, Liyuan; Li, Liangxing; Gao, Chongkai; Li, Ning

    2015-07-25

    A new biomembrane-mimetic liquid chromatographic method using a C8 stationary phase and phosphatidylcholine-modified (PC-modified) microemulsion mobile phase was used to estimate unionized and ionized drugs lipophilicity expressed as an n-octanol/water partition coefficient (logP and logD). The introduction of PC into sodium dodecyl sulfate (SDS) microemulsion yielded a good correlation between logk and logD (R(2)=0.8). The optimal composition of the PC-modified microemulsion liquid chromatography (PC-modified MELC) mobile phase was 0.2% PC-3.0% SDS-6.0% n-butanol-0.8% ethyl acetate-90.0% water (pH 7.0) for neutral and ionized molecules. The interactions between the analytes and system described by this chromatographic method is more similar to biological membrane than the n-octanol/water partition system. The result in this paper suggests that PC-modified MELC can serve as a possible alternative to the shake-flask method for high-throughput unionized and ionized drugs lipophilicity determination and simulation of biological processes. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Study of high performance liquid chromatography fingerprint for traditional chinese medicine yigongningxue oral liquid

    International Nuclear Information System (INIS)

    Feng, C.; Ruan, J.L.

    2010-01-01

    A simple, reliable and accurate fingerprinting method has been developed for quality control of YiGongNingXue oral liquid. The separation was done by reverse phase chromatography using Zorbax eclipse XDB C/sub 18/ column (250mmx4.6mmi. d. with 5.0 micro m particle size) and detection at 246nm. Methanol (5-95 % in 70 min)-buffer solution (water-phosphorica cid, pH 3.0, 95-0 percent in 70 min) as mobile phase for linear gradient elution. The flow rate was 1.0 mLmin/sup 1/ and the column temperature was at 25 deg. C. The similarity of 20 batches of YGNX oral liquid was more than 90 %. Also 15 common peaks of chromatogram have been detected, ten of them were identified by comparing fingerprint chromatogram with reference substances The HPLC fingerprint can be used to control the quality of YiGongNingXue oral liquid. (author)

  4. On-line radiometry in high-performance liquid chromatography using a storage loop

    International Nuclear Information System (INIS)

    Nieuwkerk, H.J. van.

    1987-01-01

    Difficulty with radiometric chromatography is that large measured volumes are required for accurate detections, but large volumes cause band widening and thus poor separation capacity. The solution proposed here is based on the use of a capillary storage loop for temporary residence of the column eluate. To avoid back mixing, the liquid is interspaced with a second, immiscible, phase ('spacer') so as to form a sequence of small 10-50 μl separate segments. This train passes the on-line radiometric detector to obtain a first scan of the chromatogram, called the 'direct' measurement. It then reaches the storage loop. The usual on-line UV measurement is done in the same run. The eluate cum spacer is, at a later stage, pumped through the detector at a greatly reduced flow rate to obtain the 'reversed' or 'delayed' measurement. Beta-detection is based on liquid scintillation. Accordingly, the (organic) liquid scintillant is used as the 'spacer'. In most cases it is possible to extract the analyte from the aqueous eluate into the organic phase during transport to and storage in the loop. This ensures a high counting efficiency. If the analyte is non-extractable, the counting efficiency is considerably less but the number of counts collected is still sufficient for quantitative assay, due to the extended counting time. The report is divided into 5 chapters, each dealing with a radiometric HPCL separation making use of this method. 36 figs.; 124 refs.; 16 tables

  5. [Determination of gossypol in edible vegetable oil with high performance liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Zhang, Wenhua; Huang, Chaoqun; Xie, Wen; Shen, Li

    2014-06-01

    A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the determination of gossypol in edible vegetable oil. The sample was extracted with ethyl alcohol by vortex-excited oscillation. The extract was cleaned up by 0.22 microm filter membrane and centrifuged for 5 min at 4 000 r/min after standing in a fridge at 4 degrees C for 30 min. The compound was separated on a C18 column (100 mm x 2.1 mm, 3.5 microm) with acetonitrile and 1% (v/v) formic acid aqueous solution as mobile phase. The detection of gossypol was carried out by LC-MS/MS with positive electrospray ionization under multiple reaction monitoring (MRM) mode using external standard method. The limits of quantification (S/N > 10) of gossypol in edible vegetable oil was 1 mg/kg. The recoveries were from 87.4% to 100% at the spiked levels of 1, 2, 200 mg/kg of gossypol in edible vegetable oil with the relative standard deviations (RSDs) between 3.9% and 12.2%. The method, with high sensitivity, good precision and high recovery, was suitable for the confirmation and quantification of gossypol residue in edible vegetable oil.

  6. Parallel segmented outlet flow high performance liquid chromatography with multiplexed detection

    International Nuclear Information System (INIS)

    Camenzuli, Michelle; Terry, Jessica M.; Shalliker, R. Andrew; Conlan, Xavier A.; Barnett, Neil W.; Francis, Paul S.

    2013-01-01

    Graphical abstract: -- Highlights: •Multiplexed detection for liquid chromatography. •‘Parallel segmented outlet flow’ distributes inner and outer portions of the analyte zone. •Three detectors were used simultaneously for the determination of opiate alkaloids. -- Abstract: We describe a new approach to multiplex detection for HPLC, exploiting parallel segmented outlet flow – a new column technology that provides pressure-regulated control of eluate flow through multiple outlet channels, which minimises the additional dead volume associated with conventional post-column flow splitting. Using three detectors: one UV-absorbance and two chemiluminescence systems (tris(2,2′-bipyridine)ruthenium(III) and permanganate), we examine the relative responses for six opium poppy (Papaver somniferum) alkaloids under conventional and multiplexed conditions, where approximately 30% of the eluate was distributed to each detector and the remaining solution directed to a collection vessel. The parallel segmented outlet flow mode of operation offers advantages in terms of solvent consumption, waste generation, total analysis time and solute band volume when applying multiple detectors to HPLC, but the manner in which each detection system is influenced by changes in solute concentration and solution flow rates must be carefully considered

  7. A simple subcritical chromatographic test for an extended ODS high performance liquid chromatography column classification.

    Science.gov (United States)

    Lesellier, Eric; Tchapla, Alain

    2005-12-23

    This paper describes a new test designed in subcritical fluid chromatography (SFC) to compare the commercial C18 stationary phase properties. This test provides, from a single analysis of carotenoid pigments, the absolute hydrophobicity, the silanol activity and the steric separation factor of the ODS stationary phases. Both the choice of the analytical conditions and the validation of the information obtained from the chromatographic measurements are detailed. Correlations of the carotenoid test results with results obtained from other tests (Tanaka, Engelhard, Sander and Wise) performed both in SFC and HPLC are discussed. Two separation factors, calculated from the retention of carotenoid pigments used as probe, allowed to draw a first classification diagram. Columns, which present identical chromatographic behaviors are located in the same area on this diagram. This location can be related to the stationary phase properties: endcapping treatments, bonding density, linkage functionality, specific area or silica pore diameter. From the first classification, eight groups of columns are distinguished. One group of polymer coated silica, three groups of polymeric octadecyl phases, depending on the pore size and the endcapping treatment, and four groups of monomeric stationary phases. An additional classification of the four monomeric groups allows the comparison of these stationary phases inside each group by using the total hydrophobicity. One hundred and twenty-nine columns were analysed by this simple and rapid test, which allows a comparison of columns with the aim of helping along their choice in HPLC.

  8. Parallel segmented outlet flow high performance liquid chromatography with multiplexed detection

    Energy Technology Data Exchange (ETDEWEB)

    Camenzuli, Michelle [Australian Centre for Research on Separation Science (ACROSS), School of Science and Health, University of Western Sydney (Parramatta), Sydney, NSW (Australia); Terry, Jessica M. [Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences, Deakin University, Geelong, Victoria 3216 (Australia); Shalliker, R. Andrew, E-mail: r.shalliker@uws.edu.au [Australian Centre for Research on Separation Science (ACROSS), School of Science and Health, University of Western Sydney (Parramatta), Sydney, NSW (Australia); Conlan, Xavier A.; Barnett, Neil W. [Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences, Deakin University, Geelong, Victoria 3216 (Australia); Francis, Paul S., E-mail: paul.francis@deakin.edu.au [Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences, Deakin University, Geelong, Victoria 3216 (Australia)

    2013-11-25

    Graphical abstract: -- Highlights: •Multiplexed detection for liquid chromatography. •‘Parallel segmented outlet flow’ distributes inner and outer portions of the analyte zone. •Three detectors were used simultaneously for the determination of opiate alkaloids. -- Abstract: We describe a new approach to multiplex detection for HPLC, exploiting parallel segmented outlet flow – a new column technology that provides pressure-regulated control of eluate flow through multiple outlet channels, which minimises the additional dead volume associated with conventional post-column flow splitting. Using three detectors: one UV-absorbance and two chemiluminescence systems (tris(2,2′-bipyridine)ruthenium(III) and permanganate), we examine the relative responses for six opium poppy (Papaver somniferum) alkaloids under conventional and multiplexed conditions, where approximately 30% of the eluate was distributed to each detector and the remaining solution directed to a collection vessel. The parallel segmented outlet flow mode of operation offers advantages in terms of solvent consumption, waste generation, total analysis time and solute band volume when applying multiple detectors to HPLC, but the manner in which each detection system is influenced by changes in solute concentration and solution flow rates must be carefully considered.

  9. Purification of a 166mHo solution by successive high-performance liquid chromatography and gravitational chromatography for half-life determination

    International Nuclear Information System (INIS)

    Florence Gueguen; Helene Isnard; Carole Bresson; Celine Caussignac; Guillaume Stadelmann; Anthony Nonell; Sebastien Mialle; Karsten Kossert; Frederic Chartier

    2014-01-01

    A methodology to purify a 166m Ho solution has been developed by a combination of activity and mass concentration measurements in order to further determine the 166m Ho half-life. The isobaric interference at m/q ≃ 166 requires Ho purification from non-natural Er with a high purification degree due to the large amount of Ho as opposed to Er. The Ho/Er separation was achieved using high-performance liquid chromatography on a semi-preparative column followed by purification on gravitational chromatography. The efficiency of the separation was evaluated after precise determination of the Er isotopic composition. The purification methodology enabled to separate Ho from Er. (author)

  10. Separation of three anthraquinone glycosides including two isomers by preparative high-performance liquid chromatography and high-speed countercurrent chromatography from Rheum tanguticum Maxim. ex Balf.

    Science.gov (United States)

    Chen, Tao; Li, Hongmei; Zou, Denglang; Liu, Yongling; Chen, Chen; Zhou, Guoying; Li, Yulin

    2016-08-01

    Anthraquinone glycosides, such as chrysophanol 1-O-β-d-glucoside, chrysophanol 8-O-β-d-glucoside, and physion 8-O-β-d-glucoside, are the accepted important active components of Rheum tanguticum Maxim. ex Balf. due to their pharmacological properties: antifungal, antimicrobial, cytotoxic, and antioxidant activities. However, an effective method for the separation of the above-mentioned anthraquinone glycosides from this herb is not currently available. Especially, greater difficulty existed in the separation of the two isomers chrysophanol 1-O-β-d-glucoside and chrysophanol 8-O-β-d-glucoside. This study demonstrated an efficient strategy based on preparative high-performance liquid chromatography and high-speed countercurrent chromatography for the separation of the above-mentioned anthraquinone glycosides from Rheum tanguticum Maxim.ex Balf. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Qualitative and Quantitative Analysis of Volatile Components of Zhengtian Pills Using Gas Chromatography Mass Spectrometry and Ultra-High Performance Liquid Chromatography.

    Science.gov (United States)

    Liu, Cui-Ting; Zhang, Min; Yan, Ping; Liu, Hai-Chan; Liu, Xing-Yun; Zhan, Ruo-Ting

    2016-01-01

    Zhengtian pills (ZTPs) are traditional Chinese medicine (TCM) which have been commonly used to treat headaches. Volatile components of ZTPs extracted by ethyl acetate with an ultrasonic method were analyzed by gas chromatography mass spectrometry (GC-MS). Twenty-two components were identified, accounting for 78.884% of the total components of volatile oil. The three main volatile components including protocatechuic acid, ferulic acid, and ligustilide were simultaneously determined using ultra-high performance liquid chromatography coupled with diode array detection (UHPLC-DAD). Baseline separation was achieved on an XB-C18 column with linear gradient elution of methanol-0.2% acetic acid aqueous solution. The UHPLC-DAD method provided good linearity (R (2) ≥ 0.9992), precision (RSD components, protocatechuic acid, ferulic acid, and ligustilide, in 13 batches of ZTPs, which is suitable for discrimination and quality assessment of ZTPs.

  12. Preparation and evaluation of surface-bonded tricationic ionic liquid silica as stationary phases for high-performance liquid chromatography.

    Science.gov (United States)

    Qiao, Lizhen; Shi, Xianzhe; Lu, Xin; Xu, Guowang

    2015-05-29

    Two tricationic ionic liquids were prepared and then bonded onto the surface of supporting silica materials through "thiol-ene" click chemistry as new stationary phases for high-performance liquid chromatography. The obtained columns of tricationic ionic liquids were evaluated respectively in the reversed-phase liquid chromatography (RPLC) mode and hydrophilic interaction liquid chromatography (HILIC) mode, and possess ideal column efficiency of 80,000 plates/m in the RPLC mode with naphthalene as the test solute. The tricationic ionic liquid stationary phases exhibit good hydrophobic and shape selectivity to hydrophobic compounds, and RPLC retention behavior with multiple interactions. In the HILIC mode, the retention and selectivity were evaluated through the efficient separation of nucleosides and bases as well as flavonoids, and the typical HILIC retention behavior was demonstrated by investigating retention changes of hydrophilic solutes with water volume fraction in mobile phase. The results show that the tricationic ionic liquid columns possess great prospect for applications in analysis of hydrophobic and hydrophilic samples. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. Fundamental and practical studies on high-performance liquid affinity chromatography of biopolymers with novel stationary phases

    Energy Technology Data Exchange (ETDEWEB)

    Bacolod, M.D.

    1992-01-01

    Rigid microparticulate stationary phases having surface-bound metal chelating functions were developed and evaluated in high performance metal chelate affinity chromatography of proteins. Silica- and polystyrene-divinylbenzene-based metal chelate sorbents were produced in wide pore and in non-porous type of column packings. A major effort has been placed on development of non-porous highly crosslinked polystyrene-divinylbenzene (PSDVB). These PSDVB microparticles were produced by a two-step swelling polymerization, and exhibited excellent mechanical strength over a wide range of flow-rates and composition used in high performance liquid chromatography (HPLC). Simple and reproducible hydrophilic coatings were developed for the surface modification of hydrophobic PSDVB supports. A tetradentate metal chelating ligand, ethylenediamine-N, N[prime]-diacetic acid (EDDA), was covalently bound to the surface of the various supports. Sorbents having iminodiacetic acid (IDA) metal chelating functions were also evaluated. The hydrophilic character and surface coverage of various stationary phases were assessed chromatographically. Studies concerning the effects of eluent pH as well as the nature and concentration of salts on retention and selectivity with different metal chelate stationary phases having various immobilized metal ions were carried out. Elution schemes were developed for rapid separation of proteins in metal chelate affinity chromatography. EDDA stationary phases in metal forms can be viewed as complementary to IDA stationary phases since they afforded different selectivity and retentivity toward proteins. Hydrophilic PSDVB could be functionalized with IDA or EDDA metal chelating ligands or lectins. The non-porous metal chelate stationary phases afforded rapid separation of proteins by the development of multiple gradient systems, which permitted higher column peak capacity, enabling the separation of a greater number of proteins in a single chromatographic run.

  14. Simple Quantification of Pentosidine in Human Urine and Plasma by High-Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Ji Sang Lee

    2017-01-01

    Full Text Available Pentosidine is an advanced glycation end-product (AGE and fluorescent cross-link compound. A simple high-performance liquid chromatographic (HPLC method was developed for the detection and quantification of pentosidine in human urine and plasma. The mobile phase used a gradient system to improve separation of pentosidine from endogenous peaks, and chromatograms were monitored by fluorescent detector set at excitation and emission wavelengths of 328 and 378 nm, respectively. The retention time for pentosidine was 24.3 min and the lower limits of quantification (LLOQ in human urine and plasma were 1 nM. The intraday assay precisions (coefficients of variation were generally low and found to be in the range of 5.19–7.49% and 4.96–8.78% for human urine and plasma, respectively. The corresponding values of the interday assay precisions were 9.45% and 4.27%. Accuracies (relative errors ranged from 87.9% to 115%. Pentosidine was stable in a range of pH solutions, human urine, and plasma. In summary, this HPLC method can be applied in future preclinical and clinical evaluation of pentosidine in the diabetic patients.

  15. Simultaneous determination of ethinyl estradiol and drospirenone in oral contraceptive by high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Viviane Benevenuti Silva

    2013-09-01

    Full Text Available A simple, rapid, economical and reliable high performance liquid chromatographic method has been developed and successfully applied in simultaneous determination of ethinyl estradiol and drospirenone in coated tablets. The HPLC method was performed on a LiChroCART® 100RP column (125x4 mm i.d., 5 µm with acetonitrile:water 50:50 (v/v as mobile phase, pumped at a flow rate of 1.0 mL.min-1. The fluorescence detection for ethinyl estradiol was made at λex= 280 nm and λem= 310 nm and a UV detection for drospirenone was made at 200 nm. The elution time for ethinyl estradiol and drospirenone were 4.0 and 5.7 min, respectively. The method was validated in accordance to USP 34 guidelines. The proposed HPLC method presented advantages over reported methods and is suitable for quality control assays of ethinyl estradiol and drospirenone in coated tablets.

  16. Determination of the Thermodegradation of deoxyArbutin in Aqueous Solution by High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Chih-Chien Lin

    2010-10-01

    Full Text Available Tyrosinase is the key and rate-limiting enzyme responsible for the conversion of tyrosine into melanin. Competitive inhibition of tyrosinase enzymatic activity results in decreased or absent melanin synthesis by melanocytes in human skin. DeoxyArbutin (4-[(tetrahydro-2H-pyran-2-yloxy]phenol, a novel skin whitening agent, was synthesized through the removal of hydroxyl groups from the glucose side-chain of arbutin. DeoxyArbutin not only shows greater inhibition of tyrosinase activity but is also safer than hydroquinone and arbutin. Hence, deoxyArbutin is a potential skin whitening agent for cosmetics and depigmenting drugs; however, stability of this compound under some conditions remains a problem. The lack of stability poses developmental and practical difficulties for the use of deoxyArbutin in cosmetics and medicines. Improving the thermostability of deoxyArbutin is an important issue for its development. In this research, we established an analytical procedure to verify the amount of deoxyArbutin in solutions using a high performance liquid chromatographic (HPLC method. The results indicate that this novel skin whitening agent is a thermolabile compound in aqueous solutions. Additionally, the rate constant for thermodegradation (k and the half-life (t1/2 of deoxyArbutin were determined and can be used to understand the thermodegradation kinetics of deoxyArbutin. This information can aid in the application of deoxyArbutin for many future uses.

  17. [Simultaneous determination of four common nonprotein nitrogen substances in urine by high performance liquid chromatography].

    Science.gov (United States)

    Ma, Yuhua; Huang, Dongqun; Zhang, Rui; Xu, Shiru; Feng, Shun

    2013-11-01

    A high performance liquid chromatographic (HPLC) method was proposed to simultaneously determine four common nonprotein nitrogen substances, including creatine (Cr), creatinine (Cn), uric acid (Ua) and pseudouridine (Pu) in urine. After proteins being removed by acetone precipitation method, freeze drying and redissolving, the urine samples were analyzed by HPLC. Chromatographic separation was performed on a Waters RP18 Column (150 mm x 4.60 mm, 3.5 microm) in gradient elution mode using 10.0 mmol/L KH2PO4 solution (pH 4.78) and acetonitrile as mobile phases at a flow rate of 0.8 mL/min. The samples were detected at 220 nm. Rapid separation was achieved within 7 min. Under the optimized conditions, good linearities of four common nonprotein nitrogen substances were obtained in the range of 0.1-250 mg/L. The detection limits were 9.31 (Cr), 26.19 (Cn), 4.70 (Ua), an 6.30 (Pu) microg/L and the recoveries were in the range of 81%-111% with the relative standar deviations of 0.23%-2.78% (n = 3). The results demonstrate that this method is simple, rapid and accurate with good reproducibility, and can provide early diagnosis and preliminary judgment for type 2 diabetes mellitus (T2DM) patients with renal damage.

  18. Determination of modafinil in plasma and urine by reversed phase high-performance liquid-chromatography.

    Science.gov (United States)

    Schwertner, Harvey A; Kong, Suk Bin

    2005-03-09

    Modafinil (Provigil) is a new wake-promoting drug that is being used for the management of excessive sleepiness in patients with narcolepsy. It has pharmacological properties similar to that of amphetamine, but without some of the side effects associated with amphetamine-like stimulants. Since modafinil has the potential to be abused, accurate drug-screening methods are needed for its analysis. In this study, we developed a high-performance liquid-chromatographic procedure (HPLC) for the quantitative analysis of modafinil in plasma and urine. (Phenylthio)acetic acid was used as an internal standard for the analysis of both plasma and urine. Modafinil was extracted from urine and plasma with ethyl acetate and ethyl acetate-acetic acid (100:1, v/v), respectively, and analyzed on a C18 reverse phase column with methanol-water-acetic acid (500:500:1, v/v) as the mobile phase. Recoveries from urine and plasma were 80.0 and 98.9%, respectively and the limit of quantitation was 0.1 microg/mL at 233 nm. Forty-eight 2-h post-dose urine samples from sham controls and from individuals taking 200 or 400 mg of modafinil were analyzed without knowledge of drug administration. All 16-placebo urine samples and all 32 2-h post-dose urine samples were correctly classified. The analytical procedure is accurate and reproducible and can be used for therapeutic drug monitoring, pharmacokinetic studies, and drug abuse screening.

  19. [Determination of acyclovir in mouse plasma and tissues by reversed-phase high performance liquid chromatography].

    Science.gov (United States)

    Xu, Y; Zhou, S W; Tang, J L; Huang, L Q

    2001-11-01

    The aim of this study was to establish an high performance liquid chromatographic method for determining acyclovir (ACV) concentration in mouse plasma and tissues. A solution of 0.25 mL 60 g/L perchloric acid and 0.25 mL acetonitrile was added into 0.2 mL plasma or 0.2 g tissues to precipitate proteins. Following centrifugation, the supernatant obtained was injected into a reversed-phase column. Operating conditions were Hypersil ODS column(250 mm x 4.6 mm i.d., 5 microns), methanol-water-acetic acid(1:99:0.5, volume ratio) solution as mobile phase at a flow rate of 1.5 mL/min, UV detection at 252 nm. The detection limit of ACV concentration in plasma was 20 micrograms/L and that in tissues was 50 ng/g. The standard curves for ACV were linear in plasma and homogenate of tissues (r > 0.99). The precision of the method was good and the recoveries of ACV were higher than 97.5%. So this method is rapid, accurate and convenient for determination of ACV concentrations in plasma and tissues.

  20. [Separation and determination of eight plant hormones by reversed-phase high performance liquid chromatography].

    Science.gov (United States)

    Fang, N; Hou, S; Shao, X; He, Y; Zhao, G

    1998-09-01

    In this paper, reversed-phase high performance liquid chromatographic technique was used for the separation and determination of eight plant hormones. Methanol-water-acetic acid system was chosen as the mobile phase. The effects of different separation conditions, such as the methanol and acetic acid concentrations in mobile phase, on the retention behaviours of eight plant hormones in this system were studied. The general trends in retention behaviours could be correlated to the methanol concentration in mobile phase. The experimental results showed that the optimum separation was achieved with following gradient elution condition: 0-3 minutes, 70% (water percentage in mobile phase), 3-13 minutes, 70%-20%, 13-48 minutes, 20%. Benzene was added to be as the internal standard. Under this experimental condition, the eight plant hormones could be separated completely and detected quantitatively at 260 nm within 16 minutes. The calibration curves for the eight compounds gave linearity over a wide range. The correlation coefficients of each components were r(ZT) = 0.9971, r(GAs) = 0.9999, r(K) = 0.9997, r(BA) = 0.9995, r(IAA) = 0.9998, r(IPA) = 0.9982, r(IBA) = 0.9995 and r(NAA) = 0.9995. The method is rapid, simple and efficient. It is a suitable method for the accurate determination of gibberellic acid (GA) and alpha-naphthaleneacetic acid (alpha-NAA) in products for agricultural use.