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Sample records for rapid high-performance liquid

  1. Rapid analysis of phentolamine by high-performance liquid chromatography.

    Science.gov (United States)

    Webster, Gregory K; Lemmer, Robert R; Greenwald, Steven

    2003-02-01

    A rapid liquid chromatographic method is validated for the quantitative analysis of phentolamine. Phentolamine exists in three forms for this investigation: as a mesylate salt, hydrochloride salt, and free base. In solution, phentolamine dissociates from its salt and is chromatographed as free phentolamine. This validation confirms the analysis of each form, which is simply based upon molar mass differences encountered in weighing. As such, both the United States Pharmacopeia hydrochloride and mesylate standards are used throughout this validation to demonstrate this equivalency. The validation demonstrates that this method may be used to quantitate phentolamine, regardless of its salt form.

  2. Rapid analysis of estrogen receptor heterogeneity by chromatofocusing with high-performance liquid chromatography.

    Science.gov (United States)

    Hutchens, T W; Wiehle, R D; Shahabi, N A; Wittliff, J L

    1983-08-26

    Chromatofocusing principles have been utilized to develop a high-performance liquid chromatographic technique for the rapid and routine analysis of steroid receptor heterogeneity. Two anion-exchange columns (SynChropak AX-300 and AX-500) were compared for analytical and preparative chromatofocusing of estrogen receptor components. As many as ten different [125I]iodoestradiol-labeled binding proteins were identified in cytosols prepared from mammary gland and uterus. Estrogen receptors were well separated from other cytosolic proteins and recovery of activity routinely exceeded 90%. Parallel analyses of these cytosols to determine receptor size and shape indicated that HPLC chromatofocusing can be used effectively to study receptor isoforms with Stokes radii ranging from 30 A to greater than 70 A. In contrast to isoelectric focusing, this technique is compatible with the inclusion of a commonly used receptor-stabilizing agent, sodium molybdate. Inclusion of molybdate during chromatofocusing of molybdate-stabilized receptor allowed the identification of two acidic receptor species not previously reported.

  3. [Rapid determination of propranolol enantiomers in rat plasma by column-switching-high performance liquid chromatography].

    Science.gov (United States)

    Wu, Xiaoyu; Wang, Rong; Xie, Hua; Wang, Jianfeng; Jia, Zhengping; Zhang, Qiang; Wang, Xianhua

    2011-12-01

    A high performance liquid chromatographic (HPLC) method with column-switching was developed and validated for rapid determination of two propranolol enantiomers in rat plasma. The column of restricted-access media was used as a pre-treatment column and a Chiralcel OD-RH was used as analytical column. The plasma samples were injected directly into the pretreatment column to remove plasma protein and endogenous constituents as well as to retent the propranolol enantiomers in the column using the mobile phase of borate buffer (pH 8.5)-methanol (95:5, v/v) at the flow rate of 1.0 mL/min. Then the propranolol enantiomers were transferred to the Chiralcel OD-RH column using the mobile phase of isopropanol-ethanol-0.2 mmol/L borate buffer (pH 8.5) (30: 30: 40, v/v/v) at a flow rate of 0.8 mL/min by column-switching technology. The column-switching time was 3.0 min, the used wavelength was 293 nm and the column temperature was set at 25 degrees C. The calibration curve showed excellent linear relationship (r = 0.999 5) in the concentration range from 25 mg/L to 500 mg/L for propranolol enantiomers in plasma. The intra-day and inter-day assay precisions and accuracies were well and the relative standard deviations (RSDs) were less than 5%. The average recoveries (n = 6) of the two enantiomers at 3 spiked levels were from 97.89% to 101.56%. All the values of the method validation were within the generally accepted criteria for biological sample analysis. The results show that the method is convenient, quick, sensitive and accurate. The method was successfully applied in the determination of propranolol enantiomers in rat blood pharmacokinetics study.

  4. High Performance Liquid Chromatography

    Science.gov (United States)

    Talcott, Stephen

    High performance liquid chromatography (HPLC) has many applications in food chemistry. Food components that have been analyzed with HPLC include organic acids, vitamins, amino acids, sugars, nitrosamines, certain pesticides, metabolites, fatty acids, aflatoxins, pigments, and certain food additives. Unlike gas chromatography, it is not necessary for the compound being analyzed to be volatile. It is necessary, however, for the compounds to have some solubility in the mobile phase. It is important that the solubilized samples for injection be free from all particulate matter, so centrifugation and filtration are common procedures. Also, solid-phase extraction is used commonly in sample preparation to remove interfering compounds from the sample matrix prior to HPLC analysis.

  5. Rapid determination of succinylcholine in human plasma by high-performance liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Lagerwerf, A J; Vanlinthout, L E; Vree, T B

    1991-10-04

    A high-performance liquid chromatographic method with fluorometric detection has been developed for the determination of succinylcholine in human plasma. Succinylcholine shows fluorescence at 282 nm with an excitation at 257 nm. The assay is sensitive, reproducible and linear for concentrations ranging from 100 ng/ml to 100 micrograms/ml of succinylcholine. In a pilot study the plasma concentration-time curve showed a triphasic elimination, with half-lives of 0.4, 1.2 and 8 min, respectively. In a clinical setting, drugs commonly administered during anaesthesia did not interfere with the assay. This method provides a simple and time-saving alternative to existing methods.

  6. Rapid estimation of concentration of aromatic classes in middistillate fuels by high-performance liquid chromatography

    Science.gov (United States)

    Otterson, D. A.; Seng, G. T.

    1985-01-01

    An high performance liquid chromatography (HPLC) method to estimate four aromatic classes in middistillate fuels is presented. Average refractive indices are used in a correlation to obtain the concentrations of each of the aromatic classes from HPLC data. The aromatic class concentrations can be obtained in about 15 min when the concentration of the aromatic group is known. Seven fuels with a wide range of compositions were used to test the method. Relative errors in the concentration of the two major aromatic classes were not over 10 percent. Absolute errors of the minor classes were all less than 0.3 percent. The data show that errors in group-type analyses using sulfuric acid derived standards are greater for fuels containing high concentrations of polycyclic aromatics. Corrections are based on the change in refractive index of the aromatic fraction which can occur when sulfuric acid and the fuel react. These corrections improved both the precision and the accuracy of the group-type results.

  7. Rapid determination of trace semicarbazide in flour products by high-performance liquid chromatography based on a nucleophilic substitution reaction.

    Science.gov (United States)

    Wei, Tianfu; Li, Gongke; Zhang, Zhuomin

    2017-05-01

    Semicarbazide, a toxic food contaminant, widely exists in food products and it originates from the thermal degradation of a food additive of azodicarbonamide or a metabolite of nitrofurazone abused in meat specimens. Many previous methods for semicarbazide determination usually required expensive instruments, difficult-to-prepare monoclonal antibodies, and a long operation time. In this study, a high-performance liquid chromatography method was developed for the rapid determination of trace semicarbazide coupling with a nucleophilic substitution reaction firstly using 4-nitrobenzoyl chloride as derivatization reagent. The derivatization reaction was mild at room temperature for 1 min in neutral solution. Then, semicarbazide derivative was separated and quantified by high-performance liquid chromatography with ultraviolet detection under optimal separation conditions at λmax = 261 nm. The proposed method offered the detection limit of 1.8 μg/L and was successfully applied for the rapid determination of trace semicarbazide in flour products. Semicarbazide in positive real samples could be actually found and quantified in the range of 0.47-7.53 mg/kg. The recoveries were 76.6-119% with relative standard deviations of 0.5-9.1% (n = 3). This developed method was rapid, reliable, and convenient for the determination of trace semicarbazide in food. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Rapid determination of some psychotropic drugs in complex matrices by tandem dispersive liquid-liquid microextraction followed by high performance liquid chromatography.

    Science.gov (United States)

    Asghari, Alireza; Fahimi, Ebrahim; Bazregar, Mohammad; Rajabi, Maryam; Boutorabi, Leila

    2017-03-15

    Simple and rapid determinations of some psychotropic drugs in some pharmaceutical wastewater and human plasma samples were successfully accomplished via the tandem dispersive liquid-liquid microextraction combined with high performance liquid chromatography-ultraviolet detection (TDLLME-HPLC-UV). TDLLME of the three psychotropic drugs clozapine, chlorpromazine, and thioridazine was easily performed through two consecutive dispersive liquid-liquid microextractions. By performing this convenient method, proper sample preconcentrations and clean-ups were achieved in just about 7min. In order to achieve the best extraction efficiency, the effective parameters involved were optimized. The optimal experimental conditions consisted of 100μL of CCl4 (as the extraction organic solvent), and the pH values of 13 and 2 for the donor and acceptor phases, respectively. Under these optimum experimental conditions, the proposed TDLLME-HPLC-UV technique provided a good linearity in the range of 5-3000ngmL(-1) for the three psychotropic drugs with the correlation of determinations (R(2)s) higher than 0.996. The limits of quantification (LOQs) and limits of detection (LODs) obtained were 5.0ngmL(-1) and 1.0-1.5ngmL(-1), respectively. Also the proper enrichment factors (EFs) of 96, 99, and 88 for clozapine, chlorpromazine, and thioridazine, respectively, and good extraction repeatabilities (relative standard deviations below 9.3%, n=5) were obtained.

  9. High performance liquid chromatography method for rapid and accurate determination of homocysteine in plasma and serum

    DEFF Research Database (Denmark)

    Vester, Birte; Rasmussen, K

    1991-01-01

    Determination of homocysteine in plasma or serum for evaluation of cobalamin and folate deficiency is becoming an important diagnostic procedure. Accurate, rapid and low cost methods for measuring homocysteine are therefore required. We have improved an HPLC method and made it suitable for clinical...... application. The more important changes are the addition of an internal standard, mercaptopropionylglycine, and the use of a plasma/serum based calibration material. The method consists of the following steps: reduction of the sample with tri-n-butylphosphine, precipitation of proteins, derivatisation...... with ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulphonate, and HPLC separation followed by fluorescence detection. The linearity of the assays is established and the coefficient of variation is 3.0%. Stability studies show that blood samples must be cooled or centrifuged immediately after venipuncture...

  10. Rapid screening mitochondrial DNA mutation by using denaturing high-performance liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    Man-Ran Liu; Kai-Feng Pan; Zhen-Fu Li; Yi Wang; Da-Jun Deng; Lian Zhang; You-Yong Lu

    2002-01-01

    AIM: To optimize conditions of DHPLC and analyze theeffectiveness of various DNA polymerases on DHPLCresolution, and evaluate the sensitivity of DHPLC in themutation screening of mitochondrial DNA (mtDNA).METHODS: Two fragments of 16s gene of mitochondrial DNA(one of them F2 is a mutant fragment) and an A3243Gmutated fragment were used to analyze the UV detectionlimit and determine the minimum percentage of mutant PCRproducts for DHPLC and evaluate effects of DNApolymerases on resolution of DHPLC. Under the optimalconditions, we analyzed the mtDNA mutations from muscletissues of mitochondrial encephalomyopathy with lacticacidosis and stroke-like episodes (NELAS) and screenedblindly for variances in D-loop region of mtDNA from humangastric tumor specimen.RESULTS: Ten A3243G variants were detected in 12 cases ofMELAS, no alterations were detected in controls and theseresults were consistent with the results obtained by analysisof RFLP with Apel. We also identified 26 D-loop variances in46 cases of human gastric cancer tissues and 38 alterationsin 13 gastric cancer cell lines. The mutation of mtDNA at 80 ngPCFI products containing a minimum of 5 % mutant sequencescould be detected by using DHPLC with UV detector.Moreover, Ampli-Taq Gold polymerase was equally as good asthe proofreading DNA polymerase (e. g., Pfu) in eliminatingthe false positive produced by Taq DNA polymerases.CONCLUSION: DHPLC is a powerful, rapid and sensitivemutation screening method for mtDNA. Proofreading DNApolymerase is more suitable for DHPLC analysis than Taqpolymerase.

  11. Rapid determination of Papaver somniferum alkaloids in process streams using monolithic column high-performance liquid chromatography with chemiluminescence detection

    Energy Technology Data Exchange (ETDEWEB)

    Costin, Jason W. [School of Life and Environmental Sciences, Deakin University, Geelong, Victoria 3217 (Australia); Lewis, Simon W. [Department of Applied Chemistry, Curtin University of Technology, Perth, WA 6845 (Australia); Purcell, Stuart D. [GlaxoSmithKline, Port Fairy, Victoria 3284 (Australia); Waddell, Lucy R. [GlaxoSmithKline, Port Fairy, Victoria 3284 (Australia); Francis, Paul S. [School of Life and Environmental Sciences, Deakin University, Geelong, Victoria 3217 (Australia); Barnett, Neil W. [School of Life and Environmental Sciences, Deakin University, Geelong, Victoria 3217 (Australia)]. E-mail: barnie@deakin.edu.au

    2007-07-30

    We have combined high-performance liquid chromatography (HPLC) separations using a monolithic column with acidic potassium permanganate and tris(2,2'-bipyridyl)ruthenium(II) chemiluminescence detection in a rapid and highly sensitive method to monitor the process of extracting opiate alkaloids from Papaver somniferum. Due to the high flow rates allowed with the monolithic column and the inherent selectivity of the chemiluminescence reactions, the four predominant alkaloids - morphine, codeine, oripavine and thebaine - were determined in less than 2 min. The results obtained with numerous process samples compared favourable with those of the standard HPLC methodology. Limits of detection were 1 x 10{sup -10} M, 5 x 10{sup -10} M, 5 x 10{sup -10} M and 1 x 10{sup -9} M, for morphine, codeine, oripavine and thebaine, respectively.

  12. Rapid extraction and determination of amphetamines in human urine samples using dispersive liquid-liquid microextraction and solidification of floating organic drop followed by high performance liquid chromatography.

    Science.gov (United States)

    Ahmadi-Jouibari, Toraj; Fattahi, Nazir; Shamsipur, Mojtaba

    2014-06-01

    A novel, rapid, simple and sensitive dispersive liquid-liquid microextraction method based on the solidification of floating organic drop (DLLME-SFO) combined with high-performance liquid chromatography-ultraviolet detection (HPLC-UV) was used to determine amphetamine and methamphetamine in urine samples. The factors affecting the extraction efficiency of DLLME-SFO such as the kind and volume of the extraction and the disperser solvents, effect of concentration of K2CO3 and extraction time were investigated and the optimal extraction conditions were established. Under the optimum conditions (extraction solvent: 30.0μl 1-undecanol; disperser solvent: 300μl acetonitrile; buffer concentration: 2% (w/v) K2CO3 and extraction time: 1min), calibration curves are linear in the range of 10-3000μgl(-1) and limit of detections (LODs) are in the range of 2-8μgl(-1). The relative standard deviations (RSDs) for 100μgl(-1) of amphetamine and methamphetamine in diluted urine are in the range of 6.2-7.8% (n=7). The method was successfully applied for the determination of amphetamine and methamphetamine in the actual urine samples. The relative recoveries of urine samples spiked with amphetamine and methamphetamine are 87.8-113.2%. The obtained results show that DLLME-SFO combined with HPLC-UV is a fast and simple method for the determination of amphetamine and methamphetamine in urine.

  13. Rapid genetic diagnosis and prenatal diagnosis of spinal muscular atrophy by denaturing high-performance liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    ZHU Hai-yan; WU Ling-qian; PAN Qian; TANG Bei-sha; LIANG De-sheng; LONG Zhi-gao; DAI He-ping; XIA Kun; XIA Jia-hui

    2006-01-01

    @@ Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder1 (1in 6000 to 10 000 births) caused by mutations in the SMN1 gene at 5q13. More than 90%-98% of SMA patients show homozygous deletion of SMN1,2which has proved to be useful in the diagnosis of SMA. But it is hampered because of the existence of a highly homologous gene, SMN2.3 Based on nucleotide mismatches between SMN1 and SMN2,the following two DNA tests are usually performed:single-strand conformational polymorphism (SSCP)3and polymerase chain reaction (PCR) followed by a restriction enzyme digestion.4,5 In this study we developed a new method for rapid genetic diagnosis of SMA by denaturing high-performance liquid chromatography (DHPLC), which is based on different retention of homoduplexes and heteroduplexes in detecting the homozygous deletion of SMN1. Both genetic and prenatal diagnoses were performed successfully for a SMA family by DHPLC, which was confirmed as a rapid and effective technique for detecting the deletion of SMN1.

  14. Rapid Isolation and Determination of Flavones in Biological Samples Using Zinc Complexation Coupled with High-Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Chenghe Sun

    2016-08-01

    Full Text Available Chlorophyll-type contaminants are commonly encountered in the isolation and determination of flavones of plant aerial plant parts. Heme is also a difficult background substance in whole blood analysis. Both chlorophyll and heme are porphyrin type compounds. In this study, a rapid method for isolating flavones with 5-hydroxyl or ortho-hydroxyl groups from biological samples was developed based on the different solubilities of porphyrin-metal and flavone-metal complexes. It is important that other background substances, e.g., proteins and lipids, are also removed from flavones without an additional processing. The recoveries of scutellarin, baicalin, baicalein, wogonoside and wogonin, which are the primary constituents of Scutellaria baicalensis (skullcaps were 99.65% ± 1.02%, 98.98% ± 0.73%, 99.65% ± 0.03%, 97.59% ± 0.09% and 95.19% ± 0.47%, respectively. As a sample pretreatment procedure, this method was coupled to high-performance liquid chromatography (HPLC with good separation, sensitivity and linearity and was applied to determine the flavone content in different aerial parts of S. baicalensis and in dried blood spot samples.

  15. Rapid determination of quinolones in cosmetic products by ultra high performance liquid chromatography with tandem mass spectrometry.

    Science.gov (United States)

    Liu, Shao-Ying; Huang, Xi-Hui; Wang, Xiao-Fang; Jin, Quan; Zhu, Guo-Nian

    2014-05-01

    This study developed an improved analytical method for the simultaneous quantification of 13 quinolones in cosmetics by ultra high performance liquid chromatography combined with ESI triple quadrupole MS/MS under the multiple reaction monitoring mode. The analytes were extracted and purified by using an SPE cartridge. The limits of quantification ranged from 0.03 to 3.02 μg/kg. The precision for determining the quinolones was quinolones in real cosmetic samples.

  16. Development and comparison of two dispersive liquid-liquid microextraction techniques coupled to high performance liquid chromatography for the rapid analysis of bisphenol A in edible oils.

    Science.gov (United States)

    Liu, Shuhui; Xie, Qilong; Chen, Jie; Sun, Janzhi; He, Hui; Zhang, Xiaoke

    2013-06-21

    In this study, two novel sample extraction methods for the analysis of bisphenol A (BPA) in edible oils were developed by using liquid-liquid extraction followed by a dispersive liquid-liquid microextraction (LLE-DLLME) and reversed-phase dispersive liquid-liquid microextraction (RP-DLLME). RP-DLLME showed a superior characteristic over LLE-DLLME and other previously reported procedures because of its easy operation, short extraction time, high sensitivity, low organic solvent consumption and waste generation. The optimized extraction conditions of RP-DLLME for 1.0 g of edible oil diluted in 4 mL of n-hexane were: extractant, 100 μL 0.2 M sodium hydroxide solution (80% methanol, v/v); extraction time, 1 min; centrifugation, 3 min. The determination of BPA was carried out by high performance liquid chromatography coupled with a DAD detector. The method offered excellent linearity over a range of 0.010-0.5 μg g(-1) with a correlation coefficient of r>0.997. Intra-day and inter-day repeatability values expressed as relative standard deviation were 1.9% and 5.9%, respectively. The quantitation limit and detection limit were 6.3 and 2.5 ng g(-1). The target analyte was detected in 5 out of 16 edible oil samples. The recovery rates in real samples ranged from 89.5 to 99.7%.

  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.

  18. Magnetic ionic liquid aqueous two-phase system coupled with high performance liquid chromatography: A rapid approach for determination of chloramphenicol in water environment.

    Science.gov (United States)

    Yao, Tian; Yao, Shun

    2017-01-20

    A novel organic magnetic ionic liquid based on guanidinium cation was synthesized and characterized. A new method of magnetic ionic liquid aqueous two-phase system (MILATPs) coupled with high-performance liquid chromatography (HPLC) was established to preconcentrate and determine trace amount of chloramphenicol (CAP) in water environment for the first time. In the absence of volatile organic solvents, MILATPs not only has the excellent properties of rapid extraction, but also exhibits a response to an external magnetic field which can be applied to assist phase separation. The phase behavior of MILATPs was investigated and phase equilibrium data were correlated by Merchuk equation. Various influencing factors on CAP recovery were systematically investigated and optimized. Under the optimal conditions, the preconcentration factor was 147.2 with the precision values (RSD%) of 2.42% and 4.45% for intra-day (n=6) and inter-day (n=6), respectively. The limit of detection (LOD) and limit of quantitation (LOQ) were 0.14ngmL(-1) and 0.42ngmL(-1), respectively. Fine linear range of 12.25ngmL(-1)-2200ngmL(-1) was obtained. Finally, the validated method was successfully applied for the analysis of CAP in some environmental waters with the recoveries for the spiked samples in the acceptable range of 94.6%-99.72%. Hopefully, MILATPs is showing great potential to promote new development in the field of extraction, separation and pretreatment of various biochemical samples. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. [Rapid determination of six preservative residues in fruits and vegetables by high performance liquid chromatography using dispersive solid phase extraction].

    Science.gov (United States)

    Lin, Haidan; Zou, Zhifei; Qin, Yan; Xie, Shouxin; Chen, Yufang; Xu, Juan; Wang, Lan; Chen, Chuying

    2013-02-01

    A dispersive solid phase extraction-high performance liquid chromatographic (HPLC) method was established for the simultaneous determination of 2,4-D, thiabendazole, 2-naphthol, o-phenyl phenol, diphenyl ether and biphenyl in the fruits and vegetables. The samples were vortexed for extraction with acetonitrile containing sodium chloride and anhydrous magnesium sulfate. The acetonitrile extraction was purified by acidic alumina dispersed solid phase extraction. The analysis was performed on an Agilent TC C18 column (250 mm x 4.6 mm, 5 microm) and an ultraviolet detector at 235 nm with a mobile phase of methanol-0.02 mol/L (pH 6) potassium dihydrogen phosphate solution, at a flow rate of 1.0 mL/min with gradient elution. Good linearities were observed in the range of 0.5-20 mg/L with correlation coefficients greater than 0.99. The average recoveries of the six preservatives in fruits and vegetables were in the range of 84.2% -99. 1% at three spiked levels of 1, 2 and 10 mg/kg (n = 6) and the relative standard deviations (RSDs) were in the range of 1.67%-10.3%. The limits of quantification were 1 mg/kg. This method is simple, accurate and suitable for the determination of the six preservatives in fruits and vegetables.

  20. Rapid method for determination of glyphosate in groundwater using high performance liquid chromatography and solid-phase extraction after derivatization

    Directory of Open Access Journals (Sweden)

    Valdir Eduardo Olivo

    2015-04-01

    Full Text Available The intensive use of pesticides in agriculture has prompted researchers to develop new methods for identifying these pollutants in water. This study sought to validate a high performance liquid chromatography (HPLC method to determine the concentration of the pesticide glyphosate in groundwater samples by using solid-phase extraction (SPE filters after derivatization with chloroformate 9-fluorenylmethoxycarbonyl (FMOC-Cl. For the HPLC method, we evaluated the following main validation parameters: linearity, specificity, precision, accuracy, robustness, and limits of detection and quantification. After validation of the method, we determined the concentration of glyphosate in samples from thirteen deep, tubular wells distributed in urban and rural areas in Chapecó, SC, Brazil. The solvent used in the extraction of excess FMOC-Cl was dichloromethane and subsequently filtration was performed on C18 SPE, and injected into the chromatograph column in amino polymer with fluorescence detection. The analytical curve made in ultrapure water was linear, with a correlation coefficient of 0.99. The limits of detection and quantification were 0.24 and 0.07 µg L-1, respectively. Recovery tests in natural waters ranged from 90.37 to 101.70%. Glyphosate was detected in 5 of the thirteen wells evaluated. The highest concentration of glyphosate (6.80 µg L-1 was detected in a countryside well, near the municipal water supply. Despite the low levels of glyphosate detected in our study, any amount present in groundwater samples is worrisome, as these molecules have low ground mobility.

  1. Rapid estimation of tadalafil by reverse-phase high-performance liquid chromatography method in bulk and tablet formulation

    Directory of Open Access Journals (Sweden)

    P H Sonawane

    2013-01-01

    Full Text Available The simple, selective, precise and accurate reverse-phase high-performance liquid chromatography method was developed and validated for analysis of tadalafil in bulk and tablet dosage form. The column was Inertsil C18 (150×4.6 mm; 5 μm in isocratic mode. The mobile phase used was phosphate buffer (10 mM, pH 3.2 and acetonitrile (50:50% v/v at the flow rate of 1.0 ml/min with ultraviolet detection at 295 nm at ambient temperature. The retention time for tadalafil was found to be 4.01 min. Linearity was observed in the concentration range from 60 to 140 μg/ml for tadalafil with a correlation coefficient of (r2 0.9998. The method was validated according to International Conference on Harmonisation guidelines in terms of linearity, accuracy, precision and specificity. Hence, the proposed method can be utilized for routine quality control of tadalafil in bulk and tablet dosage form.

  2. Rapid screening for drugs of abuse in biological fluids by ultra high performance liquid chromatography/Orbitrap mass spectrometry.

    Science.gov (United States)

    Jagerdeo, Eshwar; Schaff, Jason E

    2016-08-01

    We present a UPLC(®)-High Resolution Mass Spectrometric method to simultaneously screen for nineteen benzodiazepines, twelve opiates, cocaine and three metabolites, and three "Z-drug" hypnotic sedatives in both blood and urine specimens. Sample processing consists of a high-speed, high-temperature enzymatic hydrolysis for urine samples followed by a rapid supported liquid extraction (SLE). The combination of ultra-high resolution chromatography with high resolution mass spectrometry allows all 38 analytes to be uniquely detected with a ten minute analytical run. Limits of detection for all target analytes are 3ng/mL or better, with only 0.3mL of specimen used for analysis. The combination of low sample volume with fast processing and analysis makes this method a suitable replacement for immunoassay screening of the targeted drug classes, while providing far superior specificity and better limits of detection than can routinely be obtained by immunoassay.

  3. Rapid and sensitive high performance liquid chromatographic determination of zonisamide in human serum application to a pharmacokinetic study

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    M B Majnooni

    2012-01-01

    Full Text Available An accurate and very rapid method for determination of zonisamide an antiepileptic drug, in human serum is described. The analytical procedure involves liquid-liquid extraction of the analyte and an internal standard (vanillin from human serum by ethyl acetate as extracting solvent. Chromatographic separation was achieved using a monolithic C18 analytical column and a mixture of 0.05 M phosphate buffer containing triethylamine (1 ml/l; pH 2.7 and methanol (83:17 v/v was used as the mobile phase. The detection wavelength was set at 240 nm. The calibration curve was linear over a concentration range of 0.015-6.4 μg/ml of zonisamide in human serum. The total run time of analysis was 3.5 min and the lower limits of detection and quantification were 0.005 and 0.015 μg/ml, respectively. The method validation was carried out in terms of specificity, sensitivity, linearity, precision, accuracy and stability. The validated method was applied in a randomised crossover bioequivalence study of two different zonisamide preparations in 24 healthy volunteers, and the assay was sensitive enough to measure drug levels up to 8 days following a single dose administration of zonisamide.

  4. Rapid quantitative analysis of individual anthocyanin content based on high-performance liquid chromatography with diode array detection with the pH differential method.

    Science.gov (United States)

    Wang, Huayin

    2014-09-01

    A new quantitative technique for the simultaneous quantification of the individual anthocyanins based on the pH differential method and high-performance liquid chromatography with diode array detection is proposed in this paper. The six individual anthocyanins (cyanidin 3-glucoside, cyanidin 3-rutinoside, petunidin 3-glucoside, petunidin 3-rutinoside, and malvidin 3-rutinoside) from mulberry (Morus rubra) and Liriope platyphylla were used for demonstration and validation. The elution of anthocyanins was performed using a C18 column with stepwise gradient elution and individual anthocyanins were identified by high-performance liquid chromatography with tandem mass spectrometry. Based on the pH differential method, the high-performance liquid chromatography peak areas of maximum and reference absorption wavelengths of anthocyanin extracts were conducted to quantify individual anthocyanins. The calibration curves for these anthocyanins were linear within the range of 10-5500 mg/L. The correlation coefficients (r(2)) all exceeded 0.9972, and the limits of detection were in the range of 1-4 mg/L at a signal-to-noise ratio ≥5 for these anthocyanins. The proposed quantitative analysis was reproducible with good accuracy of all individual anthocyanins ranging from 96.3 to 104.2% and relative recoveries were in the range 98.4-103.2%. The proposed technique is performed without anthocyanin standards and is a simple, rapid, accurate, and economical method to determine individual anthocyanin contents.

  5. Rapid identification of erythrocyte phospholipids in Sprague-Dawley rats by ultra high performance liquid chromatography with electrospray ionization quadrupole time-of-flight tandem mass spectrometry.

    Science.gov (United States)

    Pi, Juanjuan; Wu, Xia; Yang, Shitian; Zeng, Pingyan; Feng, Yifan

    2015-03-01

    A rapid, sensitive, and reliable approach for analyzing five kinds of erythrocyte phospholipids in Sprague-Dawley rats was provided by ultra high performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry with MassLynx(TM) MassFragment. Improving conventional high performance liquid chromatography techniques, ultra high performance liquid chromatography integrated with quadrupole time-of-flight tandem mass spectrometry offers high sensitivity and increased analytical speed by using columns packed with sub-2 μm particles (1.7 μm), which allows a faster separation to be achieved. Through this method, 83 phospholipids were tentatively characterized based on their mass spectra and tandem mass spectra, as well as by matching the in-house formula database within a mass error of 5 ppm, including 40 phosphatidylcholines, 24 phosphatidyl ethanolamines, three phosphatidylinositols, six phosphatidylserines, and ten sphingomyelins. Our present results proved that the established method could be used to qualitatively analyze complex erythrocyte phospholipids in Sprague-Dawley rats and provide a useful data base for pharmacology and phospholipidomics to seek potential biomarkers of disease prediction.

  6. Rapid Mutation Scanning of Genes Associated with Familial Cancer Syndromes Using Denaturing High-Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Deborah J. Marsh

    2001-01-01

    Full Text Available Germline mutations in tumor suppressor genes, or less frequently oncogenes, have been identified in up to 19 familial cancer syndromes including Li-Fraumeni syndrome, familial paraganglioma, familial adenomatous polyposis coli and breast and ovarian cancers. Multiple genes have been associated with some syndromes as approximately 26 genes have been linked to the development of these familial cancers. With this increased knowledge of the molecular determinants of familial cancer comes an equal expectation for efficient genetic screening programs. We have trialled denaturing highperformance liquid chromatography (dHPLC as a tool for rapid germline mutation scanning of genes implicated in three familial cancer syndromes - Cowden syndrome (PTEN mutation, multiple endocrine neoplasia type 2 (RET mutation and von Hippel-Lindau disease (VHL mutation. Thirty-two mutations, including 21 in PTEN, 9 in RET plus a polymorphism, and 2 in VHL, were analyzed using the WAVE DNA fragment analysis system with 100% detection efficiency. In the case of the tumor suppressor gene PTEN, mutations were scattered along most of the gene. However, mutations in the RET proto-oncogene associated with multiple endocrine neoplasia type 2 were limited to specific clusters or “hot spots”. The use of GC-clamped primers to scan for mutations scattered along PTEN exons was shown to greatly enhance the sensitivity of detection of mutant hetero- and homoduplex peaks at a single denaturation temperature compared to fragments generated using non-GC-clamped primers. Thus, when scanning tumor suppressor genes for germline mutation using dHPLC, the incorporation of appropriate GCclamped primers will likely increase the efficiency of mutation detection.

  7. Rapid pretreatment and determination of bisphenol A in water samples based on vortex-assisted liquid-liquid microextraction followed by high-performance liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Yang, Xiao; Diao, Chun-Peng; Sun, Ai-Ling; Liu, Ren-Min

    2014-10-01

    A method for the rapid pretreatment and determination of bisphenol A in water samples based on vortex-assisted liquid-liquid microextraction followed by high-performance liquid chromatography with fluorescence detection was proposed in this paper. A simple apparatus consisting of a test tube and a cut-glass dropper was designed and applied to collect the floating extraction drop in liquid-liquid microextraction when low-density organic solvent was used as the extraction solvent. Solidification and melting steps that were tedious but necessary once the low-density organic solvent used as extraction solvent could be avoided by using this apparatus. Bisphenol A was selected as model pollutant and vortex-assisted liquid-liquid microextraction was employed to investigate the usefulness of the apparatus. High-performance liquid chromatography with fluorescence detection was selected as the analytical tool for the detection of bisphenol A. The linear dynamic range was from 0.10 to 100 μg/L for bisphenol A, with good squared regression coefficient (r(2) = 0.9990). The relative standard deviation (n = 7) was 4.7% and the limit of detection was 0.02 μg/L. The proposed method had been applied to the determination of bisphenol A in natural water samples and was shown to be economical, fast, and convenient.

  8. Ultra high performance liquid chromatography with tandem mass spectrometry for rapid separation and identification of main constituents in Chinese herbal formula Xuan Hu Suo San.

    Science.gov (United States)

    Tang, Huaqi; Zhang, Xinyue; Luo, Shilin; Shang, Fei; Chen, Jianping; Wang, Jinghong; Duan, Xiaojie; Gao, Xiaoyan; Sun, Yikun

    2017-02-15

    Xuan Hu Suo San (XHSS) is a traditional Chinese medicine that has been extensively applied in the treatment of osteoarthritis for many years, however, its chemical composition has not yet been elucidated. Thus, a rapid, efficient, and precise method based on ultra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was applied in both positive- and negative-ion modes to rapidly separate and identify the main chemical ingredients in XHSS for the first time. Finally, with the optimised separation and detection method, a total of 57 compounds were simultaneously separated within 13 min, among which 14 compounds were confirmed by comparing retention time and MS data with reference standards and others were tentatively identified by comparing with reference literatures. This rapid and sensitive approach is highly useful for the identification and characterisation of chemical constituents, and provides fundamental and extensive information supporting further metabolic and pharmacokinetic studies of XHSS.

  9. Rapid method for the analysis of itraconazole and hydroxyitraconazole in serum by high-performance liquid chromatography

    NARCIS (Netherlands)

    Compas, D; Touw, D J; de Goede, P N

    1996-01-01

    An assay for the determination of itraconazole and its active metabolite hydroxyitraconazole in serum has been developed, using ketoconazole as internal standard. The procedure involved a one-step liquid-liquid extraction of the drug, its metabolite and the internal standard, followed by their separ

  10. A rapid monitoring method for inorganic arsenic in rice flour using reversed phase-high performance liquid chromatography-inductively coupled plasma mass spectrometry.

    Science.gov (United States)

    Narukawa, Tomohiro; Chiba, Koichi; Sinaviwat, Savarin; Feldmann, Jörg

    2017-01-06

    A new rapid monitoring method by means of high performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS) following the heat-assisted extraction was developed for measurement of total inorganic arsenic species in rice flour. As(III) and As(V) eluted at the same retention time and completely separated from organoarsenic species by an isocratic elution program on a reversed phase column. Therefore, neither ambiguous oxidation of arsenite to arsenate nor the integration of two peaks were necessary to determine directly the target analyte inorganic arsenic. Rapid injection allowed measuring 3 replicates within 6min and this combined with a quantitative extraction of all arsenic species from rice flour by a 15min HNO3-H2O2 extraction makes this the fastest laboratory based method for inorganic arsenic in rice flour.

  11. Rapid determination of alpha tocopherol in olive oil adulterated with sunflower oil by reversed phase high-performance liquid chromatography.

    Science.gov (United States)

    Bakre, S M; Gadmale, D K; Toche, R B; Gaikwad, V B

    2015-05-01

    A new method is developed to determine the presence of sunflower oil in olive oil. α-tocopherol is selected as discriminating parameter for detecting sunflower oil adulterant in olive oil. Admixtures of olive oil and sunflower oil (5 %, 10 %, 15 % and 20 % sunflower oil in olive oil) are prepared. These admixtures are analysed by reversed phase high pressure liquid chromatography coupled with fluorescence detector. The sample preparation does not require saponification or addition of antioxidant. The chromatographic system consists of a C18 column with methanol: acetonitrile (50:50) mobile phase. Fluorescence detector excitation wavelength is set at 290 nm and emission wavelength is set at 330 nm. The α tocopherol concentration increases linearly in olive oil adulterated with sunflower oil. The method is simple, selective, sensitive and is precise (RSD = 2.65 %) for α tocopherol. The present method can precisely detect 5 % sunflower oil in olive oil.

  12. Ultra high performance liquid chromatography with mass spectrometry for the rapid analysis and global characterization of multiple constituents from Zibu Piyin Recipe.

    Science.gov (United States)

    Dong, Peipei; Zhang, Lin; Zhan, Libin; Liu, Yanqiu

    2016-02-01

    Zibu Piyin Recipe is a traditional Chinese medicine that has been used for the clinical treatment of memory loss in China. However, the chemical components have not been thoroughly studied so far. To quickly identify the chemical components and understand the chemical profiles related to cognition improvement activity, ultra high performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry has been applied. The analysis was performed on an Agilent 1290 ultra-high performance liquid chromatography system with a Poroshell 120 EC-C18 column (3.0 × 150 mm, 2.7 μm) using gradient elution. Using the optimized method, 155 chemical components in Zibu Piyin Recipe were tentatively identified. Among them, 112 components in negative ion mode, 73 constituents in positive ion mode, and 30 compounds in both modes could be detected. The major components were identified from Red Ginseng, Dan shen Root, White Paeony Root, liquorice root, Radix Polygalae, Tangerine Red Epicarp, Grassleaf Sweetflag Rhizome, Indian Buead, and lotus seed. These results suggest that the established rapid and robust method will be useful for identifying multiple constituents of traditional Chinese medicine prescriptions. This method could provide helpful chemical information for further in vivo pharmacological mechanism research of Zibu Piyin Recipe and new drug development for cognition improvement.

  13. Simple and Rapid Hollow Fiber Liquid Phase Microextraction Followed by High Performance Liquid Chromatography Method for Determination of Drug-protein Binding

    Institute of Scientific and Technical Information of China (English)

    XI Guo-chen; HU Shuang; BAI Xiao-hong

    2011-01-01

    A method was established using hollow fiber-liquid phase microextraction(HF-LPME) followed by high performance liquid chromatography(HPLC) to determine the concentration of the free(unbound) drug in the solution of the drug and protein.Measurements of drug-protein binding ratios and free drug concentrations were then analyzed with the Klotz equation to determine the equilibrium binding constant and number of binding sites for drug-protein interaction.The optimized method allows one to perform the efficient extraction and separation of free drug from protein-bound drug,protein,and other interfering substances.This approach was used to characterize the binding of the anticholinergic drugs atropine sulfate and scopolamine hydrobromide to proteins in human plasma and bovine serum albumin(BSA).The results demonstrate the utility of HF-LPME method for measuring free drug concentrations in protein-drug mixtures and determining the protein binding parameters of a pharmacologically important class of drugs.

  14. Options for rapid analysis of peptides and proteins, using wide-pore, superficially porous, high-performance liquid chromatography particles with unique bonded-phase ligands.

    Science.gov (United States)

    Ricker, Robert D; Woodward, Clifford B; Forrer, Kurt; Permar, Bernard J; Chen, Wu

    2008-03-01

    The large size and complexity of many proteins constrains the reversed-phase high-performance liquid chromatography packings that are useful for their separation. Wide-pore, superficially porous, silica-based packings with solid 4.5-microm cores and a 0.25-microm porous outer layer (Poroshell) demonstrate a variety of characteristics that are beneficial for the separation of proteins. A shorter diffusion distance allows separations of large molecules at high linear velocities. This benefit over totally porous particles is clearly shown using separations of a peptide-protein standard. The structure and reduced surface area (4.5 m2/g) of these superficially porous particles simplifies interactions with its surface, resulting in improved peak shapes and resolution. Specialized bonding chemistries for low- and high-pH operation may be used to change band-spacing and achieve atypical separations. These rapid analysis options are demonstrated using protein standards and very high molecular weight glycosylated proteins including intact monoclonal antibodies, IgM, alpha2-macroglobulin, and glycophorin. In liquid chromatography-mass spectrometry analysis of a myoglobin peptide digest, bidentate-C18-bonded superficially porous packings achieve complete runs in 4 min and demonstrate an elution pattern that is unique from that of material bonded with sterically protected C18 ligands.

  15. Rapid determination of amino acids in fruits of Ziziphus jujuba by hydrophilic interaction ultra-high-performance liquid chromatography coupled with triple-quadrupole mass spectrometry.

    Science.gov (United States)

    Guo, Sheng; Duan, Jin-ao; Qian, Dawei; Tang, Yuping; Qian, Yefei; Wu, Dawei; Su, Shulan; Shang, Erxin

    2013-03-20

    In this study, a sensitive and rapid method for the simultaneous determination of free amino acids without derivatization using hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry (HILIC-MS/MS) was developed. The method was performed on an ultra-high-performance liquid chromatography (UHPLC) separation system coupled with a triple-quadrupole mass spectrometry (TQ-MS) instrument. Sufficient separation of 23 underivatized amino acids was achieved on an Acquity BEH Amide column (2.1 mm × 100 mm, 1.7 μm) in a single run of 12 min. Then the method was applied for the analysis of the free amino acids in 46 batches of Ziziphus jujuba fruits which comprised 39 cultivars from 26 cultivation regions. Multivariate statistical analysis was also used to investigate the differences in free amino acid profiles among the samples. This study showed that HILIC-UHPLC-TQ-MS is an effective technique to analyze underivatized amino acids in the food samples.

  16. Hollow fiber based affinity selection combined with high performance liquid chromatography-mass spectroscopy for rapid screening lipase inhibitors from lotus leaf.

    Science.gov (United States)

    Tao, Yi; Zhang, Yufeng; Wang, Yi; Cheng, Yiyu

    2013-06-27

    A novel kind of immobilized enzyme affinity selection strategy based on hollow fibers has been developed for screening inhibitors from extracts of medicinal plants. Lipases from porcine pancreas were adsorbed onto the surface of polypropylene hollow fibers to form a stable matrix for ligand fishing, which was called hollow fibers based affinity selection (HF-AS). A variety of factors related to binding capability, including enzyme concentration, incubation time, temperature, buffer pH and ion strength, were optimized using a known lipase inhibitor hesperidin. The proposed approach was applied in screening potential lipase bound ligands from extracts of lotus leaf, followed by rapid characterization of active compounds using high performance liquid chromatography-mass spectrometry. Three flavonoids including quercetin-3-O-β-D-arabinopyranosyl-(1→2)-β-D-galactopyranoside, quercetin-3-O-β-D-glucuronide and kaempferol-3-O-β-d-glucuronide were identified as lipase inhibitors by the proposed HF-AS approach. Our findings suggested that the hollow fiber-based affinity selection could be a rapid and convenient approach for drug discovery from natural products resources.

  17. Rapid identification of mycolic acid patterns of mycobacteria by high-performance liquid chromatography using pattern recognition software and a Mycobacterium library.

    Science.gov (United States)

    Glickman, S E; Kilburn, J O; Butler, W R; Ramos, L S

    1994-01-01

    Current methods for identifying mycobacteria by high-performance liquid chromatography (HPLC) require a visual assessment of the generated chromatographic data, which often involves time-consuming hand calculations and the use of flow charts. Our laboratory has developed a personal computer-based file containing patterns of mycolic acids detected in 45 species of Mycobacterium, including both slowly and rapidly growing species, as well as Tsukamurella paurometabolum and members of the genera Corynebacterium, Nocardia, Rhodococcus, and Gordona. The library was designed to be used in conjunction with a commercially available pattern recognition software package, Pirouette (Infometrix, Seattle, Wash.). Pirouette uses the K-nearest neighbor algorithm, a similarity-based classification method, to categorize unknown samples on the basis of their multivariate proximities to samples of a preassigned category. Multivariate proximity is calculated from peak height data, while peak heights are named by retention time matching. The system was tested for accuracy by using 24 species of Mycobacterium. Of the 1,333 strains evaluated, > or = 97% were correctly identified. Identification of M. tuberculosis (n = 649) was 99.85% accurate, and identification of the M. avium complex (n = 211) was > or = 98% accurate; > or = 95% of strains of both double-cluster and single-cluster M. gordonae (n = 47) were correctly identified. This system provides a rapid, highly reliable assessment of HPLC-generated chromatographic data for the identification of mycobacteria. PMID:8195387

  18. [Rapid screening and confirming carcinogenic banned azo colorants in textiles by high performance liquid chromatography-linear ion trap/orbitrap high-resolution mass spectrometry].

    Science.gov (United States)

    Yun, Huan; Liu, Xin; Wang, Jing; Yan, Hua; Cui, Fengyun; Zhang, Zhaohui

    2013-09-01

    A method of high performance liquid chromatography-linear ion trap/orbitrap highresolution mass spectrometry (HPLC-LTP/Orbitrap MS) was ued to screen and confirm-banned azo colorants in textiles rapidly. The analytes were reduced to carcinogenic aromatic amines with sodium dithionite in citrate buffer solution. The reduced solution was extracted bydiatomite, and loadd onto an Acquity UPLC BEH C18 column (50 mm x 2.1 MM. 1.7 microm) with a gradient elution of methanol and 0.1% (v/v) methane acid aqueous solution, and finally detected by linear ion trap/orbitrap high-resolution mass spectrometry in positive ESI mode. In mass spectrometry method, the MS spectrum of high-resolution and the collision induced dissociation (CID) spectrum of data-dependent scan mode were used for screening analysis and conformation, respectively. The calibration curves showed a good linearity in the range of 0.05 -2.00 mg/b, and the correlation coefficients (r) were higher than 0.99. By detecting spiked samples, the limits of quantification were 0.08 mg/kg for all the residues and the recoveries were in the range of 65.5% - 111.5% with the relative standard deviations (RSDs) between 0.87% and 2.49%. The results indicate that the method is simple, rapid, sensitive and suitable for the qualitative and quantitative analysis of carcinogenic aromatic amines in textiles.

  19. Aptamer-affinity column clean-up coupled with ultra high performance liquid chromatography and fluorescence detection for the rapid determination of ochratoxin A in ginger powder.

    Science.gov (United States)

    Yang, Xihui; Kong, Weijun; Hu, Yichen; Yang, Meihua; Huang, Luqi; Zhao, Ming; Ouyang, Zhen

    2014-04-01

    Aptamers are single-stranded oligonucleotides with high affinity and specificity and are widely used in targets separation and enrichment. Here, an aptamer-affinity column (AAC) was firstly prepared in-house through a covalent immobilization strategy. Then, ochratoxin A (OTA) in ginger powder was absorbed and enriched using the new aptamer-based clean-up technology for the first time, and was further analyzed by ultra high performance liquid chromatography with fluorescence detection. After optimization, the average recoveries for blank samples spiked with OTA at 5, 15, and 45 μg/kg ranged from 85.36 to 96.83%. Furthermore, the AAC exhibited a similar accuracy as an immunoaffinity column to clean up OTA in ginger powder. Above all, it exhibited better reusability, twice that of the immunoaffinity column, had lower toxicity and cost, and took less time. Of 25 contaminated ginger powder samples, OTA contamination levels ranged from 1.51 to 4.31 μg/kg, which were lower than the European Union (EU) regulatory limits. All the positive samples were further confirmed by ultra-fast LC with MS/MS. In conclusion, the method of clean-up based on the AAC coupled to ultra-HPLC with fluorescence detection was rapid, specific, and sensitive for the quantitative analysis of OTA in a complex matrix.

  20. Combination of multiplex PCR with denaturing high-performance liquid chromatography for rapid detection of Mycobacterium genus and simultaneous identification of the Mycobacterium tuberculosis complex.

    Science.gov (United States)

    Chen, Ru; Gao, Xiao-Bo; Liu, Zhi-Hui; Shen, Xiao-Bing; Guo, Ai-Zhen; Duan, Yan-Yu; Liu, Zhi-Ling; Wu, Xiao-Wei; Zhu, Dao-Zhong

    2013-09-01

    A new assay with the combination of multiplex polymerase chain reaction and denaturing high-performance liquid chromatography analysis was developed for simultaneous detection of Mycobacterium genus and identification of the Mycobacterium tuberculosis complex (MTC). Targeting at genus-specific 16S rRNA sequence of Mycobacterium and specific insertion elements IS6110 and IS1081 of MTC, the assay was validated with 84 strains covering 23 mycobacteria species and 30 strains of non-mycobacteria species. No cross reactivity was observed. Clinical application was carried out on 198 specimens (155 human sputum and 43 bovine tissue samples) and compared with culture. The multiplex assay detected all culture-positive (36 in number) and 35.2% (57/162) culture-negative specimens. The molecular assay was fast that could be completed within 1 h on purified DNA, with the limit of detection as 0.8-1.6 pg per reaction on DNA template. This work provided a useful laboratory tool for rapid identification of Mycobacterium and differentiation of MTC and nontuberculous mycobacteria.

  1. Sensitive and rapid analytical method for the quantification of glucosamine in human plasma by ultra high performance liquid chromatography with tandem mass spectrometry.

    Science.gov (United States)

    Yang, Wen; Zheng, Xiaohong; Simpemba, Ernest; Ma, Pengcheng; Ding, Li

    2015-06-01

    A highly sensitive and rapid ultra high performance liquid chromatography with tandem mass spectrometry method has been developed and validated for the determination of glucosamine in human plasma using miglitol as the internal standard. Special attention was paid to achieve the high throughput and sensitivity of the established method, and the absence of a matrix effect on the analytes. The sample preparation procedure involved a simple deproteinization step. The chromatographic separation was achieved on a Waters ACQUITY HSS Cyano column using a mixture of acetonitrile/2 mM ammonium acetate solution containing 0.03% formic acid (80:20, v/v) as the mobile phase with a very short run time of 1.5 min. This method was validated over the concentration range of 10-3000 ng/mL for glucosamine. The intra- and inter-batch precision was glucosamine level in human plasma, which has not been reported in detail until now. The method was successfully applied to characterize the pharmacokinetic profile of glucosamine in healthy volunteers following a single oral administration of 750 or 1500 mg glucosamine hydrochloride.

  2. A simple and rapid ultra-high-performance liquid chromatography-tandem mass spectrometry method to determine plasma biotin in hemodialysis patients.

    Science.gov (United States)

    Yagi, Shigeaki; Nishizawa, Manabu; Ando, Itiro; Oguma, Shiro; Sato, Emiko; Imai, Yutaka; Fujiwara, Masako

    2016-08-01

    A simple, rapid, and selective method for determination of plasma biotin was developed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). After single-step protein precipitation with methanol, biotin and stable isotope-labeled biotin as an internal standard (IS) were chromatographed on a pentafluorophenyl stationary-phase column (2.1 × 100 mm, 2.7 μm) under isocratic conditions using 10 mm ammonium formate-acetonitrile (93:7, v/v) at a flow rate of 0.6 mL/min. The total chromatographic runtime was 5 min for each injection. Detection was performed in a positive electrospray ionization mode by monitoring selected ion transitions at m/z 245.1/227.0 and 249.1/231.0 for biotin and the IS, respectively. The calibration curve was linear in the range of 0.05-2 ng/mL using 300 μL of plasma. The intra- and inter-day precisions were all biotin concentrations in hemodialysis patients. Copyright © 2016 John Wiley & Sons, Ltd.

  3. A Simple, Rapid and Reliable Method to Determine Imipramine and Desipramine in Mouse Serum Using Ultra-High-Performance Liquid Chromatography-Quadrupole-Time-of-Flight Mass Spectrometry.

    Science.gov (United States)

    Zhao, Jing; Shin, Yujin; Chun, Kwang-Hoon; Yoon, Hye-Ran; Lee, Jeongmi

    2016-04-01

    A rapid and sensitive ultra-high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometric (UHPLC-Q-TOF-MS) method was developed for quantification of imipramine, one of the most widely used tricyclic antidepressants, and desipramine, an active metabolite of imipramine, in mouse serum. The developed method included a simple protein precipitation with acetonitrile in 50 μL of serum and analyte separation on an Acquity UPLC BEH C18 column using a gradient elution of acetonitrile with 0.1% formic acid and 20 mM ammonium formate. As a result, the entire analysis time was imipramine and desipramine, and calibration curves were linear over the concentration range of 5.0-1,000.0 and 5.0-250.0 ng mL(-1) for imipramine and desipramine, respectively. Intraday precisions at three levels were 2.2-3.6 and 1.7-4.2% for imipramine and desipramine, respectively, whereas interday precisions were 2.6-5.0 and 2.0-8.4% for imipramine and desipramine, respectively. Accuracy ranged between 93.6 and 106.6% for imipramine and 94.1 and 106.4% for desipramine. Absolute recovery was 96.0-97.6% for imipramine and 87.0-99.5% for desipramine. Finally, the described method was applied to mice administered with imipramine, demonstrating the suitability for quantification of imipramine and desipramine for therapeutic drug monitoring or bioequivalence studies.

  4. Molecularly imprinted polymer cartridges coupled on-line with high performance liquid chromatography for simple and rapid analysis of human insulin in plasma and pharmaceutical formulations.

    Science.gov (United States)

    Moein, Mohammad Mahdi; Javanbakht, Mehran; Akbari-adergani, Behrouz

    2014-04-01

    In this paper, a novel method is described for automated determination of human insulin in biological fluids using principle of sequential injection on a molecularly imprinted solid-phase extraction (MISPE) cartridge as a sample clean-up technique combined with high performance liquid chromatography (HPLC). The water-compatible molecularly imprinted polymers (MIPs) were prepared using methacrylic acid as a functional monomer, ethylene glycol dimethacrylate as a cross-linker, chloroform as a porogen and insulin as a template molecule. The imprinted polymers were then employed as the solid-phase extraction sorbent for on-line extraction of insulin from human plasma samples. To achieve the best condition, influential parameters on the extraction efficiency were thoroughly investigated. Rapid and simple analysis of the hormone was successfully accomplished through the good selectivity of the prepared sorbent coupled with HPLC. Limits of detection (LOD) and quantification (LOQ) of 0.2 ng mL(-1), 0.7 ng mL(-1), and 0.03 ng mL(-1), 0.1 ng mL(-1) were obtained in plasma and urine respectively. The obtained data exhibited the great recoveries for extraction of insulin from human plasma and pharmaceutical samples, higher than 87%.

  5. Rapid determination of PEGylated liposomal doxorubicin and its major metabolite in human plasma by ultraviolet–visible high-performance liquid chromatography

    Science.gov (United States)

    Chin, David L.; Lum, Bert L.; Sikic, Branimir I.

    2014-01-01

    A high-performance liquid chromatographic method was developed for the quantification of doxorubicin derived from PEGylated liposomal doxorubicin (Doxil) and its major metabolite in human plasma. This method utilizes Triton X-100 to disperse the liposome, followed by a protein precipitation step with 5-sulfosalicylic acid. Analytes in the resultant supernatant are separated on a Discovery RP amide C16 column (250×3 mm I.D., 5 µm) using an isocratic elution with a mobile phase consisting of 0.05 M sodium acetate (pH 4.0) and acetonitrile (72:28). The retention times for doxorubicin and the internal standard daunorubicin were 4.8 and 10.1 min, respectively. The column eluate was monitored by UV-visible detection at 487 nm. The determination of doxorubicin was found to be linear in the range of 1.0 ng/mL to 25 µg/mL, with intra-day and inter-day coefficients of variation and percent error ≤10%. The recovery of doxorubicin from plasma was >69.3%, with a liposomal dispersion efficiency of >95.7%. Our analytical method for free and PEGylated doxorubicin in human plasma is rapid, avoids organic extractions, and maintains sensitivity for the parent compound and its major metabolite, doxorubicinol. PMID:12361740

  6. Rapid and simple method for the determination of emodin in tartary buckwheat (Fagopyrum tataricum) by high-performance liquid chromatography coupled to a diode array detector.

    Science.gov (United States)

    Peng, Lian-Xin; Wang, Jing-Bo; Hu, Li-Xue; Zhao, Jiang-Lin; Xiang, Da-Bing; Zou, Liang; Zhao, Gang

    2013-01-30

    A simple and rapid method for determining emodin, an active factor presented in tartary buckwheat (Fagopyrum tataricum), by high-performance liquid chromatography coupled to a diode array detector (HPLC-DAD) has been developed. Emodin was separated from an extract of buckwheat on a Kromasil-ODS C(18) (250 mm × 4.6 mm × 5 μm) column. The separation is achieved within 15 min on the ODS column. Emodin can be quantified using an external standard method detecting at 436 nm. Good linearity is obtained with a correlation coefficient exceeding 0.9992. The limit of detection and the limit of quantification are 5.7 and 19 μg/L, respectively. This method shows good reproducibility for the quantification of the emodin with a relative standard deviation value of 4.3%. Under optimized extraction conditions, the recovery of emodin was calculated as >90%. The validated method is successfully applied to quantify the emodin in tartary buckwheat and its products.

  7. High-performance liquid chromatography of biopolymers

    Energy Technology Data Exchange (ETDEWEB)

    Regnier, F.E.

    1983-10-21

    The ability to separate biological macromolecules with good resolution on liquid chromatographic columns has depended on the development of suitable packing materials. In size exclusion chromatography, molecules are separated by size on the basis of differential permeation of the packing. Ion exchange, hydrophobic interaction (or reversed-phase), and affinity chromatography are all surface-mediated separation methods, although they depend on different retention mechanisms. High-performance liquid chromatographic columns designed for biopolymers offer major advantages over conventional columns in both speed and resolving power. The exponential growth of literature on the high-performance separation of peptides and proteins in particular indicates that the techniques will become the dominant form of column liquid chromatography. 92 refs., 4 figs.

  8. Rapid determination of triclosan in personal care products using new in-tube based ultrasound-assisted salt-induced liquid-liquid microextraction coupled with high performance liquid chromatography-ultraviolet detection.

    Science.gov (United States)

    Chen, Ming-Jen; Liu, Ya-Ting; Lin, Chiao-Wen; Ponnusamy, Vinoth Kumar; Jen, Jen-Fon

    2013-03-12

    This paper describes the development of a novel, simple and efficient in-tube based ultrasound-assisted salt-induced liquid-liquid microextraction (IT-USA-SI-LLME) technique for the rapid determination of triclosan (TCS) in personal care products by high performance liquid chromatography-ultraviolet (HPLC-UV) detection. IT-USA-SI-LLME method is based on the rapid phase separation of water-miscible organic solvent from the aqueous phase in the presence of high concentration of salt (salting-out phenomena) under ultrasonication. In the present work, an indigenously fabricated home-made glass extraction device (8-mL glass tube inbuilt with a self-scaled capillary tip) was utilized as the phase separation device for USA-SI-LLME. After the extraction, the upper extractant layer was narrowed into the self-scaled capillary tip by pushing the plunger plug; thus, the collection and measurement of the upper organic solvent layer was simple and convenient. The effects of various parameters on the extraction efficiency were thoroughly evaluated and optimized. Under optimal conditions, detection was linear in the concentration range of 0.4-100ngmL(-1) with correlation coefficient of 0.9968. The limit of detection was 0.09ngmL(-1) and the relative standard deviations ranged between 0.8 and 5.3% (n=5). The applicability of the developed method was demonstrated for the analysis of TCS in different commercial personal care products and the relative recoveries ranged from 90.4 to 98.5%. The present method was proven to be a simple, sensitive, less organic solvent consuming, inexpensive and rapid procedure for analysis of TCS in a variety of commercially available personal care products or cosmetic preparations.

  9. Multicomponent mixed dopant optimization for rapid screening of polycyclic aromatic hydrocarbons using ultra high performance liquid chromatography coupled to atmospheric pressure photoionization high-resolution mass spectrometry

    KAUST Repository

    Sioud, Salim

    2012-05-04

    RATIONALE To enhance the ionization efficiencies in atmospheric pressure photoionization mass spectrometry a dopant with favorable ionization energy such as chlorobenzene is typically used. These dopants are typically toxic and difficult to mix with water-soluble organic solvents. In order to achieve a more efficient and less toxic dopant, a multicomponent mixed dopant was explored. METHODS A multicomponent mixed dopant for non-targeted rapid screening of polycyclic aromatic hydrocarbons (PAHs) was developed and optimized using ultra high performance liquid chromatography (UPLC) coupled to atmospheric pressure photoionization high-resolution mass spectrometry. Various single and multicomponent mixed dopants consisting of ethanol, chlorobenzene, bromobenzene, anisole and toluene were evaluated. RESULTS Fourteen out of eighteen PAHs were successfully separated and detected at low pg/μL levels within 5 min with high mass accuracy ≤4 ppm. The optimal mixed multicomponent dopant consisted of ethanol/chlorobenzene/bromobenzene/anisole (98.975:0.1:0.9:0.025, v/v %) and it improved the limit of detection (LOD) by 2- to 10-fold for the tested PAHs compared to those obtained with pure chlorobenzene. CONCLUSIONS A novel multicomponent dopant that contains 99% ethanol and 1% mixture of chlorobenzene, bromobenzene and anisole was found to be an effective dopant mixture to ionize PAHs. The developed UPLC multicomponent dopant assisted atmospheric pressure photoionization high-resolution mass spectrometry offered a rapid non targeted screening method for detecting the PAHs at low pg/;μL levels within a 5 min run time with high mass accuracy a;circ4 ppm. Copyright © 2012 John Wiley & Sons, Ltd.

  10. Basic prinicipal of high performance liquid chromatography

    OpenAIRE

    Ivanova, Violeta

    2016-01-01

    High-performance liquid chromatography (HPLC) is an analytical technique based on the separation of molecules due to differences in their structure and/or composition. Separation is performed between two phases, mobile and stationary. The molecules in the sample have different affinities and interactions with the stationary support, leading to separation of molecules. Compounds which are longer retained at the stationary phase will elute later, compared to those which are distributed into the...

  11. A simple, rapid and validated high-performance liquid chromatography method suitable for clinical measurements of human mercaptalbumin and non-mercaptalbumin.

    Science.gov (United States)

    Yasukawa, Keiko; Shimosawa, Tatsuo; Okubo, Shigeo; Yatomi, Yutaka

    2017-01-01

    Background Human mercaptalbumin and human non-mercaptalbumin have been reported as markers for various pathological conditions, such as kidney and liver diseases. These markers play important roles in redox regulations throughout the body. Despite the recognition of these markers in various pathophysiologic conditions, the measurements of human mercaptalbumin and non-mercaptalbumin have not been popular because of the technical complexity and long measurement time of conventional methods. Methods Based on previous reports, we explored the optimal analytical conditions for a high-performance liquid chromatography method using an anion-exchange column packed with a hydrophilic polyvinyl alcohol gel. The method was then validated using performance tests as well as measurements of various patients' serum samples. Results We successfully established a reliable high-performance liquid chromatography method with an analytical time of only 12 min per test. The repeatability (within-day variability) and reproducibility (day-to-day variability) were 0.30% and 0.27% (CV), respectively. A very good correlation was obtained with the results of the conventional method. Conclusions A practical method for the clinical measurement of human mercaptalbumin and non-mercaptalbumin was established. This high-performance liquid chromatography method is expected to be a powerful tool enabling the expansion of clinical usefulness and ensuring the elucidation of the roles of albumin in redox reactions throughout the human body.

  12. A novel fatty-acid-based in-tube dispersive liquid-liquid microextraction technique for the rapid determination of nonylphenol and 4-tert-octylphenol in aqueous samples using high-performance liquid chromatography-ultraviolet detection.

    Science.gov (United States)

    Shih, Hou-Kuang; Shu, Ting-Yun; Ponnusamy, Vinoth Kumar; Jen, Jen-Fon

    2015-01-07

    In this study, a novel fatty-acid-based in-tube dispersive liquid-liquid microextraction (FA-IT-DLLME) technique is proposed for the first time and is developed as a simple, rapid and eco-friendly sample extraction method for the determination of alkylphenols in aqueous samples using high-performance liquid chromatography-ultraviolet detection (HPLC-UV). In this extraction method, medium-chain saturated fatty acids were investigated as a pH-dependent phase because they acted as either anionic surfactants or neutral extraction solvents based on the acid-base reaction caused solely by the adjustment of the pH of the solution. A specially designed home-made glass extraction tube with a built-in scaled capillary tube was utilized as the phase-separation device for the FA-IT-DLLME to collect and measure the separated extractant phase for analysis. Nonylphenol (NP) and 4-tert-octylphenol (4-tOP) were chosen as model analytes. The parameters influencing the FA-IT-DLLME were thoroughly investigated and optimized. Under the optimal conditions, the detector responses of NP and 4-tOP were linear in the concentration ranges of 5-4000 μg L(-1), with correlation coefficients of 0.9990 and 0.9996 for NP and 4-tOP, respectively. The limits of detection based on a signal-to-noise ratio of 3 were 0.7 and 0.5 μg L(-1), and the enrichment factors were 195 and 143 for NP and 4-tOP, respectively. The applicability of the developed method was demonstrated for the analysis of alkylphenols in environmental wastewater samples, and the recoveries ranged from 92.9 to 107.1%. The extraction process required less than 4 min and utilized only acids, alkalis, and fatty acids to achieve the extraction. The results demonstrated that the presented FA-IT-DLLME approach is highly cost-effective, simple, rapid and environmentally friendly in its sample preparation.

  13. Isocratic Solid Phase Extraction-Liquid Chromatography (SPE-LC) Interfaced to High-Performance Tandem Mass Spectrometry for Rapid Protein Identification

    DEFF Research Database (Denmark)

    Hørning, Ole B; Kjeldsen, Frank; Theodorsen, Søren

    2008-01-01

    the isocratic solid phase extraction-liquid chromatography (SPE-LC) technology for rapid separation ( approximately 8 min) of simple peptide samples. We now extend these studies to demonstrate the potential of SPE-LC separation in combination with a hybrid linear ion trap-Orbitrap tandem mass spectrometer......Reversed-phase liquid chromatography interfaced to electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) allows analysis of very complex peptide mixtures at great sensitivity, but it can be very time-consuming, typically using 60 min, or more, per sample analysis. We recently introduced...

  14. Simultaneous and rapid determination of deoxynivalenol and its acetylate derivatives in wheat flour and rice by ultra high performance liquid chromatography with photo diode array detection.

    Science.gov (United States)

    Xu, Jiao-Jiao; Zhou, Jian; Huang, Bai-Fen; Cai, Zeng-Xuan; Xu, Xiao-Min; Ren, Yi-Ping

    2016-06-01

    A simple and reliable method of ultra high performance liquid chromatography coupled with photo-diode array detection has been proposed for the simultaneous determination of deoxynivalenol and its acetylated derivatives in wheat flour and rice, especially focusing on the optimization of sample extraction, cleanup, and chromatographic separation conditions. Sample pretreatment consisted of a first step using a quick, easy, cheap, effective, rugged, and safe based extraction procedure and a subsequent cleanup step based on solid-phase extraction. The method was extensively validated in wheat flour and rice, obtaining satisfactory analytical performance with good linearity (R(2) ≥ 0.999), acceptable recoveries (80.0-104.4%), and repeatability (RSDs 1.3-10.7%). The limits of detection (21.7-57.4 μg/kg) and quantitation (72.3-191.4 μg/kg) for deoxynivalenols were lower than those usually permitted by various countries' legislation in these food matrices. The method was applied to 34 wheat and rice samples. The results were further compared with results of ultra high performance liquid chromatography with electrospray ionization tandem mass spectrometry.

  15. Rapid High Performance Liquid Chromatography Determination and Optimization of Extraction Parameters of the α-Asarone Isolated from Perilla frutescens L.

    Science.gov (United States)

    Hwang, Seung Hwan; Kwon, Shin Hwa; Kang, Young-Hee; Lee, Jae-Yong; Lim, Soon Sung

    2017-02-10

    Response surface methodology (RSM), based on a central composite design, was used to determine the best liquid-to-raw material ratio (10:3-15 mL/g), extraction time (1-3 h), and ethanol concentration (50%-100%) for maximum content of α-asarone from Perilla frutescens (PF) extract. Experimental values of α-asarone were 9.51-46.36 mg/g; the results fitted a second-order quadratic polynomial model and correlated with the proposed model (R2 > 0.9354). The best conditions were obtained with extraction time of 1.76 h, liquid-to-raw material ratio of 10:13.5 mL/g, and ethanol concentration of 90.37%. Under these conditions, the model predicted extraction content of 40.56 mg/g, while experimental PF content of α-asarone was 43.84 mg/g dried plant. Optimized conditions determined for maximum content of α-asarone were similar to the experimental range. Experimental values agreed with those predicted, thus validating and indicating suitability of both the model and the RSM approach for optimizing extraction conditions. In addition, a reliable, reproducible and accurate method for the quantitative determination of α-asarone by High Performance Liquid Chromatography (HPLC) analysis was developed with limit of detection (LOD), limit of quantitation (LOQ) values of 0.10 and 0.29 µg/mL and excellent linearity (R2 > 0.9999).

  16. Sensitive and Rapid Quantification of Felodipine by High-performance Liquid Chromatography-tandem Mass Spectrometry(HPLC-MS/MS) and Its Pharmacokinetics in Healthy Chinese Volunteers

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    To investigate the pharmacokinetics of felodipine in the plasma of healthy Chinese volunteers, 30 healthy volunteers received a single oral dose of 5 mg of extended release felodipine tablets. The felodipine was extracted from the matrix with a liquid-liquid extract procedure and analyzed by high-performance liquid chromatography-tandem mass spectrometry in the multiple reaction monitoring(MRM) mode using an electrospray ion source with positive ion detection. The method was validated over a felodipine concentration range of 0.05-10.00 ng/mL in human plasma. Its main pharmacokinetic parameters values were: ρmax=(1.67±0.84) ng/mL, occurring at (3.93±2.49) h; the plasma elimination half-life: (23.08±9.48) h and the area under the plasma concentration versus time curve: (29.94±14.39) ng·h/mL. The validation results demonstrated that this method showed a satisfactory precision and accuracy across the calibration range. The procedure involved minimal drug administration, sample preparation, and a 2.5-min chromatographic run time. It was well suited to clinical studies of the drug involving large numbers of samples.

  17. Development and validation of a rapid high performance liquid chromatography - photodiode array detection method for estimation of a bioactive compound wedelolactone in extracts of Eclipta alba

    Directory of Open Access Journals (Sweden)

    Satyanshu Kumar

    2013-03-01

    Full Text Available Following optimization of extraction, separation and analytical conditions, a rapid, sensitive and simple reverse-phase high performance liquid chromatography-photo diode array (HPLC-PDA method has been developed for the identification and quantification of wedelolactone in different extracts of Eclipta alba. The separation of wedelolactone was achieved on a C18 column using the solvent system consisting of a mixture of methanol: water: acetic acid (95: 5: 0.04 as a mobile phase in isocratic elution mode followed by photo diode array detection at 352 nm. The developed method was validated as per the guidelines of the International Conference on Harmonization (ICH. Calibration curve presented good linear regression (r²>0.998 within the test range and the maximum relative standard deviation (RSD, % values for intra-day assay were found to be 0.15, 1.30 and 1.1 for low (5 µg/mL, medium (20 µg/mL and high (80 µg/mL concentrations of wedelolactone. For inter-day assay the maximum RSD (% values were found to be 2.83, 1.51 and 2.06 for low, medium and high concentrations, respectively. Limit of detection (LOD and limit of quantification (LOQ were calculated to be 2 and 5 µg/mL respectively. Analytical recovery of wedelolactone was greater than 95%. Wedelolactone in different extracts of Eclipta alba was identified and quantified using the developed HPLC method. The validated HPLC method allowed precise quantitative analysis of wedelolactone in Eclipta. alba extracts.Desenvolveu-se método rápido, sensível e simples de Cromatografia Líquida de Alta Eficiência em fase reversa, utilizando-se arranjo de fotodiodo (HPLC-PDA, visando à separação, extração e às condições analíticas para a identificação e quantificação de wedelolactona em diferentes extratos de Eclipta alba. A separação de wedelolactona foi efetuada por meio de uma coluna C18, utilizando mistura de metanol:água:ácido acético (95:5:0.04 como fase móvel, em sistema de

  18. High Performance Liquid Chromatography/Video Fluorometry. Part I. Instrumentation.

    Science.gov (United States)

    1981-09-30

    High Performance Liquid Chromatography /Video...PERIOD COVERED High Performance Liquid Chromatography /Video .. / Fluorometry. Part I. Instrumentation. . Interim/ echnicaliepart,. 6. PERFORMING ORG...34Entered SECURITY CLASSIFICATION OF THIS OlAGE (When Data Entered) II1| III I I I I E I II ... .. High Performance Liquid Chromatography

  19. Rapid and sensitive determination of benzo[a]pyrene in black ginseng using fluorescence detector and high performance liquid chromatography-tandem mass spectrometry

    Science.gov (United States)

    Cho, Hyun-jeong; Kim, Hye-jin; Son, Byeong-cheol; Jo, Dong-keun; Cho, Byung-lim

    2013-05-01

    Black ginseng is produced by steaming a ginseng root followed by drying repeatedly 9 times during the process and it is changed to be black color, so it is known that a black ginseng has more contents of saponins than red ginseng. However a fake black ginseng which is produced to be black color at high temperature in a short period of time generate carcinogenic benzo[a]pyrene(BaP) through the process. In this year, maximum residue level(MRL) for BaP was established to 2 ug/kg in black ginseng and more sensitive method was developed to quantitatively analyze the BaP by high performance liquid chromatography (HPLC) coupling with florescence detector and tandem mass spectrometry (atmospheric pressure chemical ionization-MS/MS). Chromatographic separation was performed on a Supelcosil™ LC-PAH column (3 μm, 3 mm x 50 mm). Mobile phase A was water and mobile phase B was acetonitrile. BaP was exactly separated from other 15 polycyclic aromatic hydrocarbons (PAHs) which have been selected as priority pollutants by the US Environmental Protection Agency (EPA). Linearity of detection was in the range of 0.2~20 μg/kg and limit of detection (LOD) for BaP was lower than 0.1 μg/kg, limit of quantification (LOQ) was 0.2 μg/kg. The recovery of Bap was 92.54%+/-6.3% in black ginseng.

  20. Rapid modified QuEChERS method for pesticides detection in honey by high-performance liquid chromatography UV-visible

    Directory of Open Access Journals (Sweden)

    Elisabetta Bonerba

    2014-05-01

    Full Text Available The extensive use of pesticides in agriculture plays an important role in bees die-off and allows the presence of residues in hive products, particularly in honey. An accurate and reliable analytical method, based on QuEChERS extractive technique, has been developed for the quantitative determination by high-performance liquid chromatography UV-visible detector of 5 pesticides (Deltamethrin, Dimethoate, Imidacloprid, Acetamiprid, Chlorfenvinphos in honey. The method, according to Commission Directive 2002/63/EC and Regulation 882/2004/EC, provided excellent results with respect to linearity (correlation coefficient up to 0.993, limits of detection and quantification (0.005 and 0.01 μg/mL for Dimethoate, Deltamethrin and Chlorfenvinphos; 0.02 and 0.05 μg/mL for Acetamiprid and Imidacloprid, recovery values (86.4 to 96.3%, precision and relative expanded uncertainty of a measurement, demonstrating the conformity of the this method with the European directives. The proposed method was applied to 23 samples of Apulian honey. None of the investigated pesticides was detected in these samples.

  1. Magnetic mixed hemimicelles solid-phase extraction coupled with high-performance liquid chromatography for the extraction and rapid determination of six fluoroquinolones in environmental water samples.

    Science.gov (United States)

    Wang, Li; Yuan, Qiuxiang; Liang, Guoxi; Shi, Longbiao; Zhan, Qian

    2015-03-01

    In this study, a mixed hemimicelle solid-phase extraction method based on Fe3 O4 nanoparticles coated with sodium dodecyl sulfate was applied for the preconcentration and fast isolation of six fluoroquinolones in environmental water samples before high-performance liquid chromatography determination. The main factors affecting the extraction efficiency of the analytes, such as amount of surfactant, amount of Fe3 O4 nanoparticles, extraction time, sample volume, sample pH, ionic strength, and desorption conditions, were investigated and optimized. The method has detection limits from 0.05 to 0.1 ng/mL and good linearity (r ≥ 09948) in the range 0.1-200 ng/mL depending on the fluoroquinolone. The enrichment factor is ∼200. The recoveries (at spiked levels of 1, 5, and 50 ng/mL) are in the range of 79-120%. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Rapid and sensitive method for the determination of acetaldehyde in fuel ethanol by high-performance liquid chromatography with UV-Vis detection

    Energy Technology Data Exchange (ETDEWEB)

    Saczk, Adelir Aparecida; Okumura, Leonardo Luiz; Firmino de Oliveira, Marcelo; Boldrin Zanoni, Maria Valnice; Ramos Stradiotto, Nelson [Instituto de Quimica, UNESP, Quitandinha, Araraquara, SP (Brazil)

    2005-04-01

    A high-performance liquid chromatography (HPLC) method for the determination of acetaldehyde in fuel ethanol was developed. Acetaldehyde was derivatized with 0.900 mL 2,4-dinitrophenylhydrazine (DNPHi) reagent and 50 {mu}L phosphoric acid 1 mol L{sup -1} at a controlled room temperature of 15 C for 20 min. The separation of acetaldehyde-DNPH (ADNPH) was carried out on a Shimadzu Shim-pack C{sub 18} column, using methanol/LiCl{sub (aq)} 1.0 mM (80/20, v/v) as a mobile phase under isocratic elution and UV-Vis detection at 365 nm. The standard curve of ADNPH was linear in the range 3-300 mg L{sup -1} per injection (20 {mu}L) and the limit of detection (LOD) for acetaldehyde was 2.03 {mu}g L{sup -1}, with a correlation coefficient greater than 0.999 and a precision (relative standard deviation, RSD) of 5.6% (n=5). Recovery studies were performed by fortifying fuel samples with acetaldehyde at various concentrations and the results were in the range 98.7-102%, with a coefficient of variation (CV) from 0.2% to 7.2%. Several fuel samples collected from various gas stations were analyzed and the method was successfully applied to the analysis of acetaldehyde in fuel ethanol samples. (orig.)

  3. A novel ultra-high performance liquid chromatography method for the rapid determination of β-lactoglobulin as heat load indicator in commercial milk samples.

    Science.gov (United States)

    Boitz, Lisa I; Fiechter, Gregor; Seifried, Reinhold K; Mayer, Helmut K

    2015-03-20

    The level of undenatured acid-soluble β-lactoglobulin can be used as an indicator to assess the heat load applied to liquid milk, thus further allowing the discrimination between milk originating from different thermal production processes. In this work, a new UHPLC method for the rapid determination of bovine β-lactoglobulin in 1.8min only (total runtime 3min) is presented using simple UV detection at 205nm. Separation selectivity for possibly co-eluting other major whey proteins (bovine serum albumin, lactoferrin, α-lactalbumin, immunoglobulin G) was verified, and the method validated for the analysis of liquid milk samples regarding linearity (20-560μg/mL, R(2)>0.99), instrumentation precision (RSDsESL) milk, quite diverse levels were determined ranging from ∼100 up to 4000mg/L, thus clearly illustrating variable applied heat loads and impacts on the "nativeness" of milk essentially due to the fact that the production technologies used for ESL milk may differ significantly, and are currently not regulated in the EU.

  4. Development and validation of a rapid column-switching high-performance liquid chromatographic method for the determination of Lamotrigine in human serum.

    Science.gov (United States)

    Brunetto, María del Rosario; Contreras, Yaritza; Delgado, Yelitza; Gallignani, Máximo; Estela, José Manuel; Martin, Víctor Cerdà

    2009-07-01

    This study describes a simple and sensitive column-switching high-performance liquid chromatographic method with UV detection for the determination of Lamotrigine in 50 microL of serum. After solid-phase extraction of Lamotrigine on an Oasis HLB extraction precolumn (20 x 3.9 mm; dp: 25 microm), chromatographic separation was achieved at 30 degrees C on a Chromolith RP-18e column (50 mm x 4.6 mm i.d.) using a solution of 20% acetonitrile in 15 mM phosphate buffer (pH 7.0) as the mobile phase, at a flow-rate of 2.0 mL/min. The eluant was detected at 215 nm. The retention time for Lamotrigine was 1.28 min. The total analysis time was ca. 5 min. However, the overlap of sample preparation, analysis, and reconditioning of the precolumn increased the overall sample throughput to one injection every 3 min. The method was validated for system suitability, linearity, precision, accuracy, robustness, and limit of quantitation. The linearity of the calibration lines, expressed by the linear correlation coefficient, was better than 0.9996. Recovery studies achieved from Lamotrigine spiked plasma samples showed values greater than 93%, demonstrating the excellent extraction efficiency of the precolumn. Intra- and inter-day precision were generally acceptable; the coefficient of variation was < 2.3% in all cases. The detection limits for Lamotrigine at a signal-to-noise ratio of 3 was 0.002 microg/mL when a sample volume of 50 microL was injected. However, it was possible to enhance the sensitivity further by injecting larger volumes, up to 200 microL. The method was shown to be robust and the results were within the acceptable range. The method was successfully applied to the determination of Lamotrigine in human serum samples of patients submitted to Lamotrigine therapy.

  5. A rapid and simple high-performance liquid chromatography method for the determination of human plasma levofloxacin concentration and its application to bioequivalence studies.

    Science.gov (United States)

    Zhou, Zhi-Ling; Yang, Min; Yu, Xi-Yong; Peng, Huai-Yan; Shan, Zhi-Xin; Chen, Shu-Zhen; Lin, Qiu-Xiong; Liu, Xiao-Ying; Chen, Tie-Feng; Zhou, Shu-Feng; Lin, Shu-Guang

    2007-10-01

    A high-performance liquid chromatography method with fluorescence detection (HPLC-FLD) for the determination of levofloxacin in human plasma is described. Neutralized with phosphate buffer (pH 7.0), the sample (0.1 mL) was extracted with dichlormethane (1 mL). After voltex-mixing and centrifuged at 3000g for 6 min at 4 degrees C, the upper aqueous layer was aspirated using a micro vacuum pump and the organic layer was directly transferred to a clean test tube without pipetting. The organic solvent was evaporated and the residues were reconstituted with the mobile phase. Levofloxacin and terazosin (internal standard, IS) were chromatographically separated on a C(18) column with a mobile phase containing phosphate buffer (pH 3.0, 10 mm), acetonitrile and triethylamine (76:24:0.076, v/v/v) at a flow rate of 1 mL/min. The analytes were detected using fluorescence detection at an excitation and emission wavelength of 295 and 440 nm, respectively. The linear range of the calibration curves was 0.0521-5.213 microg/mL for levofloxacin with a lower limit of quantitation (0.0521 microg/mL). The retention times of levofloxacin and terazosin were 2.5 and 3.1 min, respectively. Within- and between-run precision was less than 12 and 11%, respectively. Accuracy ranged from -6.3 to 4.5%. The recovery ranged from 86 to 89% at the concentrations of 0.0521, 0.5213 and 5.213 microg/mL. The present HPLC-FLD method is sensitive, efficient and reliable. The method described herein has been successfully used for the pharmacokinetic and bioequivalence studies of a levofloxacin formulation product after oral administration to healthy Chinese volunteers.

  6. Simple and rapid method on High Performance Liquid Chromatography for simultaneous determination of benzylpenicillin potassium, streptomycin sulphate and related substances in Ascomicin – a veterinary use ointment

    Directory of Open Access Journals (Sweden)

    Neagu Maria

    2015-06-01

    Full Text Available A new simple, rapid, accurate and precise High – Performance Liquid Chromatography (HPLC method for determination of benzylpenicillin potassium and streptomycin sulphate in Ascomicin ointment was developed and validated. The method can be used for the detection and quantification of known and unknown impurities and degradation products in this pharmaceutical product during routine analysis and also for stability studies in view of its capability to separate degradation products. The method was validated for accuracy, precision, specificity, robustness and quantification limits according to ICH Guidelines. The estimation of benzylpenicillin potassium and streptomycin sulphate was done by Waters HPLC 2695. The chromatographic conditions comprised a reverse-phased C18 column (5 µm particle size, 250 mm×4.6 mm i.d. with a mobile phase consisting of a mixture of solution in water containing 0.025 M of sodium phosphate dibasic and 0.02 of sodium hexansulfonate adjusted to pH 6.0 with 22.5 g/lsolution of phosphoric acid and acetonitrile in gradient elution. The flow rate was 0.8 ml/min. Standard curves were linear over the concentration range of 5.00 µg/ml to 5.00 mg/ml for streptomycin sulphate and 3.26 µg/ml to 3.26 mg/ml for benzylpenicillin potassium. Statistical analyses proved the method was precise, reproducible, selective, specific and accurate for analysis of benzylpenicillin potassium, streptomycin sulphate and related substances.

  7. A fast reverse-phase high performance liquid chromatographic tandem mass spectrometry assay for the quantification of clindamycin in plasma and saliva using a rapid resolution package.

    Science.gov (United States)

    Catena, Esther; Perez, Guiomar; Sadaba, Belen; Azanza, Jose Ramón; Campanero, Miguel Angel

    2009-11-01

    A new method for the quantitative analysis of clindamycin in human plasma and saliva by liquid chromatography/electrospray ionisation tandem mass spectrometry (LC/ESI-MS/MS) has been developed using a rapid resolution C18 column (2.1 mm x 30 mm x 3.5 microm). A simple deproteinization procedure was applied to the samples before analysis. Multiple reaction monitoring (MRM) mode of precursor-product ion transitions for clindamycin (425.1/126.1) and the internal standard, lincomycin (407.2/126.0) was used. Chromatographic separation was achieved at 0.6 ml/min in less than 1.5 min, with improved peak resolution and sensitivity between drug and internal standard. The assay exhibited a linear dynamic range between 0.05 and 15.0 microg/ml and gave a determination coefficient of 0.991 or better. The limit of quantification of the method was 10 ng/ml in both biological samples. Intra-day and inter-day precision ranged from 7.5% to 11.5%. Good accuracy was observed for both the intra-day and inter-day assays (R.S.D. below +/-4%). The suitability of the developed method for the analysis of clindamycin in plasma and saliva samples was demonstrated by the measure of clindamycin in samples taken up to 6h after oral and intravenous administration of this drug in infectious patients.

  8. Rapid Determination of Dopamine and Its Metabolites During in vivo Cerebral Microdialysis by Routine High Performance Liquid Chromatography With Electrochemical Detection

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Objective To determine dopamine and its metabolites during in vivo cerebral microdialysis by routine high performance liquid chromatography with electrochemical detection. Methods Microdialysis probes were placed into the right striatum of Wistar rat brains and perfused with Ringer's solution at a rate of 1.5 μL/min. A reverse phase HPLC with electrochemistry was used to assay DA, DOPAC, and HVA after cerebral microdialysates were collected every 20 minutes from awake and freely moving rats. In order to identify the reliability of this method, its selectivity, linear range, precision and accuracy were tested and the contents of DA, DOPAC, and HVA in rat microdialysates were determined. Results The standard curve was in good linear at the concentration ranging from 74 nmol/L to 1.5 μmol/L for DOPAC (r2= 0.9996), from 66 nmol/L to 1.3 μmol/L for DA (r2=1.0000) and from 69 nmol/L to 1.4 μmol/L for HVA (r2=0.9992). The recovery of DOPAC (0.30, 0.77, 1.49 μmol/L),DA (0.26, 0.69, 1.32 μmol/L), and HVA (0.27, 0.71, 1.37 μmol/L) was 82.00±1.70%, 104.00±4.00%, 98.70±3.10%;92.30±1.50%, 105.30±2.30%, 108.00±2.00%; 80.00±7.80%, 107.69±8.00%, and 108.66±3.10%, respectively at each concentration. Their intra-day RSD was 3.3%, 3.4%, and 2.5%, and inter-day RSD was 4.2%, 2.3%, and 5.6%, respectively.The mean extracellular concentrations of DOPAC, DA, and HVA in rat brain microdialysates were 10.7, 2.4, and 9.2 μmol/L (n=6), respectively. Conclusion The findings of our study suggested that the simple, accurate and stable method can be applied to basic researches of diseases related to monoamines neurotransmitters by cerebral microdialysis in rats.

  9. Rapid and sensitive screening of N-glycans as 9-fluorenylmethyl derivatives by high-performance liquid chromatography: a method which can recover free oligosaccharides after analysis.

    Science.gov (United States)

    Kamoda, Satoru; Nakano, Miyako; Ishikawa, Rika; Suzuki, Shigeo; Kakehi, Kazuaki

    2005-01-01

    There are a large number of labeling methods for asparagine-type oligosaccharides with fluorogenic and chromophoric reagents. We have to choose the most appropriate labeling method based on the purposes such as mass spectrometry, high-performance liquid chromatography and capillary electrophoresis. Asparagine-type glycans are released from core proteins as N-glycosylamine at the initial step of the releasing reaction when glycoamidase F is employed as the enzyme. The N-glycosylamine-type oligosaccharides thus released by the enzyme are subjected to hydrolysis or mutarotation to form free-form oligosaccharides. In the detailed studies on the enzyme reaction, we found a condition in which the released N-glycosylamine-type oligosaccharides were exclusively present at least during the course of enzyme reaction, and developed a method for in situ derivatization of the glycosylamine-type oligosaccharides with 9-fluorenylmethyl chloroformate (Fmoc-Cl). The Fmoc labeled sialo- and asialo- (or high-mannose and hybrid) oligosaccharides were successfully analyzed on an amine-bonded polymer column and amide-silica column, respectively. The present method showed approximately 5 times higher sensitivities than that using 2-aminobenzoic acid (2-AA). The separation profile was similar to that observed using 2-AA method as examined by the analyses of carbohydrate chains derived from several glycoproteins including complex-type, high-mannose type and hybrid type of N-linked oligosaccharides. The labeled oligosaccharides were stable at least for several months when stored at -20 degrees C. Furthermore, it should be emphasized that the Fmoc-derivatized oligosaccharides could be easily recovered as free reducing oligosaccharides simply by incubation with morpholine in dimethylformamide solution. We obtained a pure triantennary oligosaccharide with 3 sialic acid residues as a free reducing form from fetuin in good yield after isolation of the corresponding Fmoc oligosaccharide followed

  10. High Performance Liquid Chromatography/Video Fluorometry. Part II. Applications.

    Science.gov (United States)

    1981-09-30

    HIGH PERFORMANCE LIQUID CHROMATOGRAPHY /VIDEO FLUOROMETRY. PART...REP«T_N&:-ŗ/ High Performance Liquid Chromatography /Video Fluorometry» Part II. Applications« by | Dennis C./Shelly* Michael P./Vogarty and...Data EnlirtdJ REPORT DOCUMENTATION PAGE t. REPORT NUMBER 2 GOVT ACCESSION NO 4. T1TI.F (and Submit) lP-^fffsyva High Performance Liquid Chromatography

  11. Quantification of tryptophan in plasma by high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Renata Romanholi Pinhati

    2012-01-01

    Full Text Available A simple, rapid and selective method using high-performance liquid chromatography with ultraviolet detection (267 nm was applied for the determination of tryptophan in plasma. Separation was carried out on a C18 column (150 x 4.6 mm internal diameter in 6 min. The mobile phase consisted of 5 mM the sodium acetate and acetonitrile (92:8, v/v. The method was shown to be precise and accurate, and good recovery of analyte was achieved, characterizing the method as efficient and reliable for use in laboratory analysis.

  12. High-Performance Liquid Chromatography: An Overview

    OpenAIRE

    Tandia N; Singh SK1; Kumar N; Singh S

    2013-01-01

    Chromatography is considered extremely powerful separation technique for variety of samples. Thecommon feature of all these techniques is that the different components in a sample mixture aredistributed between two phases, one of which remains stationary while the other the mobile phase, runsthrough the interstices or over the surface of the fixed phase. The movement of the mobile phase resultsin differential migration of the sample components. High performance thin layer chromatography is an...

  13. Rapid resolution liquid chromatography-mass spectrometry and high-performance liquid chromatography-fluorescence detection for metabolism and pharmacokinetic studies of ergosta-4,6,8(14),22-tetraen-3-one.

    Science.gov (United States)

    Zhao, Ying-Yong; Qin, Xiang-Yang; Cheng, Xian-Long; Liu, Xue-Ying; Lin, Rui-Chao; Zhang, Yongmin; Li, Xiao-Ye; Sun, Xiao-Li; Sun, Wen-Ji

    2010-08-24

    Ergosta-4,6,8(14),22-tetraen-3-one (ergone) from many medicinal plants has been demonstrated to possess a variety of pharmacological activities in vivo and in vitro, including cytotoxic, diuretic and immunosuppressive activity. Metabolism and pharmacokinetic studies on rat were conducted for ergone. Rapid resolution liquid chromatography with atmospheric pressure chemical ionization tandem multi-stage mass spectrometry (RRLC-APCI-MS(n)) and high-performance liquid chromatography with fluorescence detection (HPLC-FLD) methods were applied for the identification and quantification of ergone and its metabolite from rat plasma, faeces and urine. A metabolite was identified by RRLC-DAD-APCI-MS(n): 22,23-epoxy-ergosta-4,6,8(14)-triaen-3-one (epoxyergone). The concentrations of the analyte with its metabolites were determined by HPLC-FLD at excitation wavelength of 370 nm and emission wavelength of 485 nm. The samples were deproteinized with methanol after addition of camptothecin as internal standard (IS). The analysis was performed on a Diamonsil C18 column (150 mm x 4.6 mm x 5 microm) with a mobile phase gradient consisting of methanol and water at a flow rate of 1 mL min(-1). The assay was linear over the concentration range of 42-1500, 36-7500 and 42-1500 ng mL(-1) for plasma, faecal homogenate and urine respectively. The absolute recoveries were found to be 97.0+/-1.2%, 98.1+/-0.7% and 96.6+/-1.8% for plasma, faecal homogenate and urine respectively. The intra-day and inter-day relative standard deviations (RSD) were less than 10%. The previous HPLC-MS/MS method is not affordable for most laboratories because of the specialty requirement and high equipment cost. However, the HPLC-FLD method is economic and operating simply for quantitative determination of ergone and its metabolite in rat plasma, faeces and urine. In addition, liquid chromatography coupled with ion trap multi-stage mass spectrometry is becoming a useful technique for ergone metabolite identification.

  14. Ultra-high performance liquid chromatography with linear ion trap-Orbitrap hybrid mass spectrometry combined with a systematic strategy based on fragment ions for the rapid separation and characterization of components in Stellera chamaejasme extracts.

    Science.gov (United States)

    Wang, Zhixin; Qu, Yang; Wang, Li; Zhang, Xiaozhe; Xiao, Hongbin

    2016-04-01

    Stellera chamaejasme, a famous toxic herb, has been used in traditional Chinese medicine to treat various diseases. For decades, increasing attention in modern pharmacological studies has been drawn to S. chamaejasme because of its potential anti-tumor, anti-virus, and anti-fungus activities. However, due to the intrinsic complexity of chemical constitutes, hardly any investigations formed an overall recognition for the chemical profiles of this herb. In this study, a rapid and sensitive ultra-high performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometry method was developed to characterize the chemical components of S. chamaejasme extracts. Based on optimized ultra-high performance liquid chromatography and mass spectrometry conditions and systematic fragment ions-based strategy, a total of 47 components including flavones, diterpenes, coumarins, and lignans were simultaneously detected and identified or tentatively identified for the first time. The MS(n) fragmentation patterns of all the characterized compounds in positive or negative electrospray ionization modes were also explored and summarized. These results provided essential data for further pharmacological research on S. chamaejasme. Moreover, the method was demonstrated to be an efficient tool for rapid qualitative analysis of secondary metabolites from natural resources.

  15. Direct analysis of six antibiotics in wastewater samples using rapid high-performance liquid chromatography coupled with diode array detector: a chemometric study towards green analytical chemistry.

    Science.gov (United States)

    Vosough, Maryam; Rashvand, Masoumeh; Esfahani, Hadi M; Kargosha, Kazem; Salemi, Amir

    2015-04-01

    In this work, a rapid HPLC-DAD method has been developed for the analysis of six antibiotics (amoxicillin, metronidazole, sulfamethoxazole, ofloxacine, sulfadiazine and sulfamerazine) in the sewage treatment plant influent and effluent samples. Decreasing the chromatographic run time to less than 4 min as well as lowering the cost per analysis, were achieved through direct injection of the samples into the HPLC system followed by chemometric analysis. The problem of the complete separation of the analytes from each other and/or from the matrix ingredients was resolved as a posteriori. The performance of MCR/ALS and U-PLS/RBL, as second-order algorithms, was studied and comparable results were obtained from implication of these modeling methods. It was demonstrated that the proposed methods could be used promisingly as green analytical strategies for detection and quantification of the targeted pollutants in wastewater samples while avoiding the more complicated high cost instrumentations.

  16. Rapid quantitation of avenanthramides in oat-containing products by high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (HPLC-TQMS).

    Science.gov (United States)

    Xie, Zhuohong; Mui, Tracy; Sintara, Marsha; Ou, Boxin; Johnson, Jodee; Chu, YiFang; O'shea, Marianne; Kasturi, Prabhakar; Chen, Yumin

    2017-06-01

    Avenanthramides (AVNs) are a family of nitrogen-containing phenolic compounds produced in oat; AVN 2c, 2p, and 2f are the three major members. An LC-MS/MS method was developed, with the limit of detection (LOD) and the limit of quantitation (LOQ) being, respectively, 0.29ng/mL and 1.96ng/mL for AVN 2c, 0.24ng/mL and 0.60ng/mL for AVE 2p, and 0.42ng/mL and 2.2ng/mL for AVN 2f. The method was validated in oat-containing hot cereal and snack bar samples. The recovery of AVN 2c, 2p, and 2f from these two oat products was 95-113%, and the relative standard deviations ranged from 5% to 9%. This method was used to evaluate oat products and raw oat samples. The effects of location and variety on AVN composition were investigated. The method presented here provides a novel and rapid tool to quantitate the abundance of AVN 2c, 2p, and 2f in oat-containing products.

  17. Development and validation of a rapid ultra-high performance liquid chromatography method for the assay of benzalkonium chloride using a quality-by-design approach.

    Science.gov (United States)

    Mallik, Rangan; Raman, Srividya; Liang, Xiaoli; Grobin, Adam W; Choudhury, Dilip

    2015-09-25

    A rapid robust reversed-phase UHPLC method has been developed for the analysis of total benzalkonium chloride in preserved drug formulation. A systematic Quality-by-Design (QbD) method development approach using commercial, off the shelf software (Fusion AE(®)) has been used to optimize the column, mobile phases, gradient time, and other HPLC conditions. Total benzalkonium chloride analysis involves simple sample preparation. The method uses gradient elution from an ACE Excel 2 C18-AR column (50mm×2.1mm, 2.0μm particle size), ammonium phosphate buffer (pH 3.3; 10mM) as aqueous mobile phase and methanol/acetonitrile (85/15, v/v) as the organic mobile phase with UV detection at 214nm. Using these conditions, major homologs of the benzalkonium chloride (C12 and C14) have been separated in less than 2.0min. The validation results confirmed that the method is precise, accurate and linear at concentrations ranging from 0.025mg/mL to 0.075mg/mL for total benzalkonium chloride. The recoveries ranged from 99% to 103% at concentrations from 0.025mg/mL to 0.075mg/mL for total benzalkonium chloride. The validation results also confirmed the robustness of the method as predicted by Fusion AE(®).

  18. Rapid high performance liquid chromatographic method for determination of clarithromycin in human plasma using amperometric detection: application in pharmacokinetic and bioequivalence studies.

    Science.gov (United States)

    Foroutan, Seyed Mohsen; Zarghi, Afshin; Shafaati, Alireza; Madadian, Babak; Abolfathi, Farshid

    2013-01-01

    A rapid, sensitive and reproducible HPLC method using amperometric detector was developed and validated for the analysis of clarithromycin in human plasma. The separation was achieved on a monolithic silica column (MZ- C8 125×4.0 mm) using acetonitrile-methanol-potassium dihydrogen phosphate buffer (40:6:54,v/v), with pH of 7.5, as the mobile phase at a flow rate of 1.5 mL/min. The assay enables the measurement of clarithromycin for therapeutic drug monitoring with a minimum quantification limit of 20 ng/mL. The method involves simple, protein precipitation procedure and analytical recovery was complete. The calibration curve was linear over the concentration range of 0.1-6 μg/mL. The coefficients of variation for inter-day and intra-day assay were found to be less than 6%. This method was used in bioequivalency and pharmacokinetic studies of the test (generic) product 2 × 500 mg clarithromycin tablets, with respect to the reference product.

  19. New rapid methods for determination of total LAS in sewage sludge by high performance liquid chromatography (HPLC) and capillary electrophoresis (CE)

    Energy Technology Data Exchange (ETDEWEB)

    Villar, M. [Department of Analytical Chemistry, Faculty of Chemistry, University of Seville, c/Profesor Garcia Gonzalez s/n, 41012 Seville (Spain); Callejon, M. [Department of Analytical Chemistry, Faculty of Chemistry, University of Seville, c/Profesor Garcia Gonzalez s/n, 41012 Seville (Spain)], E-mail: mochon@us.es; Jimenez, J.C.; Alonso, E.; Guiraum, A. [Department of Analytical Chemistry, Faculty of Chemistry, University of Seville, c/Profesor Garcia Gonzalez s/n, 41012 Seville (Spain)

    2009-02-23

    Linear alkylbenzene sulfonates (LAS) are the most common synthetic anionic surfactant used in domestic and industrial detergents, with a global production of 2.4 x 10{sup 6} t year{sup -1}. After use and disposal, LAS may enter the environment by one of the several routes, including by direct discharge to surface water or discharge to water from sewage treatment plants. Sewage treatment plants break down LAS only partly: some of them remain in effluent and other fraction is adsorbed in sewage solid. New and rapid methods for determination of total LAS from sewage sludge based on microwave assisted extraction and HPLC-FL and CE-DAD determination are proposed. The extraction of total LAS is carried out by using microwaves energy, an extraction time of 10 min and 5 mL of methanol. For HPLC-FL determination, mobile phase acetonitrile-water was used, comprising 60% (v/v) from 0 to 1 min and a flow rate of 1 mL min{sup -1} programmed to 100% acetonitrile between 1 and 2 min and a flow rate of 2 mL min{sup -1}. The final composition was maintained for a further 5 min. The determination of total LAS by CE-DAD was performed in a phosphate buffer (10 mM, pH 9). The separation voltage was 25 kV and the temperature of the capillary was 30 deg. C. Injections were performed in the pressure mode and the injection time was set at 12 s. The determination of total LAS is carried out in less than 5 min. The methods did not require clean-up or preconcentration steps. Detection limit for total LAS in the sludge was 3.03 mg kg{sup -1} using HPLC-FL and 21.0 mg kg{sup -1} using CE-DAD, and recoveries were >85% using both determination methods. Concentrations of total LAS obtained using both methods were compared with the sum of concentrations of homologues LAS C-10, LAS C-11, LAS C-12 and LAS C-13 obtained using microwaves assisted extraction and HPLC-FL and CE-DAD determination.

  20. 巧克力中香兰素的快速分离柱高效液相色谱法测定%Determination of Vanillin in Chocolate by Rapid Column High Performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    李海燕; 汤丹俞; 缪恩明; 贺宾; 施红林; 蔡继宝

    2009-01-01

    [Objective]The aim was to discuss the effect of using rapid column high performance liquid chromatography to determine vanillin in chocolate. [Method]The rapid column high performance liquid chromatography was used to determine vanillin in chocolate. [Result]40 ml 40% methanol were selected to extract vanillin from chocolate by ultrasonic for 10 min, then solid phrase extraction was applied for pre-separation. The vanillin was separated with ZORBAX Stable Bound (4.6×50 mm, 1.8 μm) C18 as stationary phase and methanol-water (35∶65) as mobile phase at flow-rate of 10 ml/min, and the detector was UV photodiode array detector. Under the chromatographic conditions, vanillin could reach baseline separation in 1.0 min. The sample standard recoveries were 98%-103% and the relative standard deviations were 0.46%-0.58%. The separation time could be shortened above 85% compared with conventional chromatograph. [Conclusion]The method was convenient and rapid, and the result was reliable, which provided method for rapid determination of vanillin in chocolate to some extent.%[目的]探讨快速分离柱高效液相色谱法测定巧克力中的香兰素的效果.[方法]采用快速分离柱高效液相色谱法测定巧克力中的香兰素.[结果]选用40%甲醇40 ml超声振荡10 min提取巧克力中的香兰素,然后用固相萃取预分离,以 ZORBAX Stable Bound (4.6×50 mm, 1.8 μm) C18为固定相,甲醇-水(35∶65)为流动相分离,流速为 10 ml/min,用紫外二极管矩阵检测器检测.在该色谱条件下,香兰素在1.0 min内可达到基线分离.样品的标准回收率为98%~103%,相对标准偏差为0.46%~0.58%.和常规色谱相比可缩短85%以上的分离时间.[结论]该方法简便快速,结果可靠,在一定程度上为巧克力中香兰素的快速测定提供了方法.

  1. ANALYSIS OF AMINO ACIDS BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

    Directory of Open Access Journals (Sweden)

    Qurat E Noor Baig

    2016-06-01

    Full Text Available Amino acids are the building blocks of proteins and are considered as the key precursors for the formation of hormones and low molecular weight nitrogenous substances with biological importance. Since the analysis of amino acids has been carried out for both qualitative and quantitative purposes with an aim to study their levels in the plasma concentration, the quantitative determination, in particular, also helps in the diagnosis of different diseases associated with their deficiency. This review article deals with the determination of amino acids by chromatographic methods which include ion-exchange chromatography (IEC, high performance liquid chromatography (HPLC, reverse phase-high performance liquid chromatography (RP-HPLC and ultra-performance liquid chromatography (UPLC. The review will also give an idea for the preparation of samples, derivatization methods for the analysis of amino acids (direct and indirect methods and separation of amino acids by high performance liquid chromatographic technique.

  2. Development of a Rapid and Simultaneous Detection Method for Buprenorphine, Norbuprenorphine and Naloxone in Human Plasma Using Ultra-high Performance Liquid Chromatography- tandem Mass Spectrometer with Solid-phase Extraction%Development of a Rapid and Simultaneous Detection Method for Buprenorphine, Norbuprenorphine and Naloxone in Human Plasma Using Ultra-high Performance Liquid Chromatography- tandem Mass Spectrometer with Solid-phase Extraction

    Institute of Scientific and Technical Information of China (English)

    Guo, Qiaozhen; Du, Zhenxia

    2011-01-01

    A simultaneous method was successfully established and validated for the separation and determination of bu- prenorphine (BP), its primary metabolite, nor-buprenorphine (NBP) and a proposed co-formulate, naloxone (NLX) in human plasma. The method used buprenorphine-d4 (BP-D4), nor-buprenorphine-d3 (NBP-D3), naltrexone (NTX) as internal standards (ISs). 100 μL of plasma sample fortified with the ISs was cleaned up by solid-phase extraction (SPE), and was then separated on a Waters AcquityTM BEH C18 column with gradient elution using methanol and water (containing 0.2% formic) at a flow rate of 0.25 mL·min^-1. The mass spectrometer was used for detection and was operated in the positive electrospray ionization with multiple reaction monitoring (MRM) mode. The three compounds were effectively separated in 5 min. The linear ranges of the compounds were 0.1--25, 0.25--25 and 0.05--25 ng·mL^-1 for BP, NBP and NLX, respectively, with r≥0.9935. The method had high sensitivity (the lim- its of detection were 0.02, 0.1 and 0.01 ng.mL-1 for BP, NBP and NLX, respectively) and high recoveries (≥97.6%). The result was shown to be linear and satisfactorily met current acceptance criteria for validation of bio- analytical method: intra and inter assay precisions within the required limits of ≤25% RSD. The LOQs fulfilled the LOQ requirements: precision≤25% RSD, and was fully validated according to the State Food and Drug Administration (SFDA) regulations. The results demonstrated that ultra-high performance liquid chromatography- tandem mass spectrometer (UPLC-MS/MS) with SPE was a powerful detection tool and contributed to pharmaceutical analysis in biological matrices.

  3. 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.

  4. 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...

  5. 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)

  6. Multichannel Detection in High-Performance Liquid Chromatography.

    Science.gov (United States)

    Miller, James C.; And Others

    1982-01-01

    A linear photodiode array is used as the photodetector element in a new ultraviolet-visible detection system for high-performance liquid chromatography (HPLC). Using a computer network, the system processes eight different chromatographic signals simultaneously in real-time and acquires spectra manually/automatically. Applications in fast HPLC…

  7. 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…

  8. Analysis of Tocopherols by High Performance Liquid Chromatography

    OpenAIRE

    Edison, B.

    2009-01-01

    : Gas chromatography is the key technique for organic components and also for tocopherols analysis. High performance liquid chromatography has an important role to take part in applications such as the handling of less usual samples, prevention of degradation of heat sensitive functional groups and for micro preparative purposes. Many approaches for development of improved methods are suggested, especially for reversed phase applications.

  9. 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.

  10. 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…

  11. 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 reversed-p

  12. UPLC-MS-MS法快速测定玉米中的黄曲霉毒素B1%Ultra-High Performance Liquid Chromatography Tandem Mass Spectrometry for Rapid Determination of Aflatoxin B1 in Corn

    Institute of Scientific and Technical Information of China (English)

    于成广; 黄辉

    2012-01-01

    采用高效液相色谱-串联质谱法检测玉米中的黄曲霉毒素B1.对样品前处理条件、净化和分离条件进行了优化,黄曲霉毒素B1检出限为0.02μg/kg,回收率为83.8%~95.7%.建立的方法简便、快速、灵敏度高,能够满足玉米中痕量黄曲霉毒素B1农残的高灵敏测定要求.%An analytical method for the determination of aflatoxin Bl in corn by ultra-high performance liquid chromatography tandem mass spectrometry was developed. Sample pretreatment methods and depuration conditions were optimized. The detection limit of aflatoxin Bl was 0. 02 μg/kg. The recoveries were 83.8% - 95.7%. The results indicated that the method developed is simple, rapid, sensitive, which is suit for the analysis of aflatoxin Bl pesticides in corn.

  13. Determination of unbound vismodegib (GDC-0449) concentration in human plasma using rapid equilibrium dialysis followed by solid phase extraction and high-performance liquid chromatography coupled to mass spectrometry.

    Science.gov (United States)

    Deng, Yuzhong; Wong, Harvey; Graham, Richard A; Liu, Wenbin; Shen, Heuy-shin; Shi, Yao; Wang, Laixin; Meng, Min; Malhi, Vikram; Ding, Xiao; Dean, Brian

    2011-07-15

    A rapid equilibrium dialysis (RED) assay followed by a solid phase extraction (SPE) high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) assay for the quantitative determination of unbound vismodegib in human plasma was developed and validated. The equilibrium dialysis was carried out using 0.3 mL plasma samples in the single-use plate RED system at 37°C for 6h. The dialysis samples (0.1 mL) were extracted using a Strata-X-C 33u Polymeric Strong Cation SPE plate and the resulting extracts were analyzed using reverse-phase chromatography and positive electrospray ionization (ESI) mass spectrometry. The standard curve, which ranged from 0.100 to 100 ng/mL for vismodegib, was fitted to a 1/x(2) weighted linear regression model. The lower limit of quantitation (LLOQ, 0.100 ng/mL) was sufficient to quantify unbound concentrations of vismodegib after dialysis. The intra-assay precision of the LC-MS/MS assay, based on the four analytical QC levels (LLOQ, low, medium and high), was within 7.7% CV and inter-assay precision was within 5.5% CV. The assay accuracy, expressed as %Bias, was within ±4.0% of the nominal concentration values. Extraction recovery of vismodegib was between 77.9 and 84.0%. The assay provides a means for accurate assessment of unbound vismodegib plasma concentrations in clinical studies.

  14. Rapid determination of thiamine, riboflavin, niacinamide, pantothenic acid, pyridoxine, folic acid and ascorbic acid in Vitamins with Minerals Tablets by high-performance liquid chromatography with diode array detector.

    Science.gov (United States)

    Jin, Pengfei; Xia, Lufeng; Li, Zheng; Che, Ning; Zou, Ding; Hu, Xin

    2012-11-01

    A simple, isocratic, and stability-indicating high-performance liquid chromatography (HPLC) method has been developed for the rapid determination of thiamine (VB(1)), niacinamide (VB(3)), pyridoxine (VB(6)), ascorbic acid (VC), pantothenic acid (VB(5)), riboflavin (VB(2)) and folic acid (VB(9)) in Vitamins with Minerals Tablets (VMT). An Alltima C(18) column (250 mm × 4.6 mm i.d., 5 μm) was used for the separation at ambient temperature, with 50mM ammonium dihydrogen phosphate (adjusting with phosphoric acid to pH 3.0) and acetonitrile as the mobile phase at the flow rate of 0.5 ml min(-1). VB(1), VB(3), VB(6), VC and VB(5) were extracted with a solution containing 0.05% phosphoric acid (v/v) and 0.3% sodium thiosulfate (w/v), and were then simultaneously analyzed by using the mobile phase of phosphate buffer-acetonitrile (95:5, v/v), while VB(2) and VB(9) were extracted with a solution containing 0.5% ammonium hydroxide solution (v/v), and were then simultaneously analyzed by using the mobile phase of phosphate buffer-acetonitrile (85:15, v/v). The detection wavelengths were 275 nm for VB(1), VB(3), VB(6), VC, 210 nm for VB(5), and 282 nm for VB(2) and VB(9). The method showed good system suitability, sensitivity, linearity, specificity, precision, stability and accuracy. All the seven water-soluble vitamins were well separated from other ingredients and degradation products. Method comparison indicated good concordance between the developed method and the USP method. The developed method was reliable and convenient for the rapid determination of VB(1), VB(3), VB(6), VC, VB(5), VB(2) and VB(9) in VMT.

  15. Temperature-Modulated Array High-Performance Liquid Chromatography

    OpenAIRE

    Premstaller, Andreas; Xiao, Wenzhong; Oberacher, Herbert; O'Keefe, Matthew; Stern, David; Willis, Thomas; Huber, Christian G.; Peter J. Oefner

    2001-01-01

    Using novel monolithic poly(styrene-divinylbenzene) capillary columns with an internal diameter of 0.2 mm, we demonstrate for the first time the feasibility of constructing high-performance liquid chromatography arrays for the detection of mutations by heteroduplex analysis under partially denaturing conditions. In one embodiment, such an array can be used to analyze one sample simultaneously at different temperatures to maximize the detection of mutations in DNA fragments containing multiple...

  16. Analysis of Tocopherols by High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    B. Edison

    2009-01-01

    Full Text Available : Gas chromatography is the key technique for organic components and also for tocopherols analysis. High performance liquid chromatography has an important role to take part in applications such as the handling of less usual samples, prevention of degradation of heat sensitive functional groups and for micro preparative purposes. Many approaches for development of improved methods are suggested, especially for reversed phase applications.

  17. A novel method for the rapid determination of polyethoxylated tallow amine surfactants in water and sediment using large volume injection with high performance liquid chromatography and tandem mass spectrometry.

    Science.gov (United States)

    Ross, Andrew R S; Liao, Xiangjun

    2015-08-19

    Polyethoxylated tallow amine (POEA) surfactants have been used in many glyphosate-based herbicide formulations for agricultural, industrial and residential weed control. The potential for release of these compounds into the environment is of increasing concern due to their toxicity towards aquatic organisms. Current methods for analysis of POEA surfactants require significant time and effort to achieve limits of quantification that are often higher than the concentrations at which biological effects have been observed (as low as 2 ng mL(-1)). We have developed a rapid and robust method for quantifying the POEA surfactant mixture MON 0818 at biologically relevant concentrations in fresh water, sea water and lake sediment using reversed phase high-performance liquid chromatography and electrospray ionization-tandem mass spectrometry. Water samples preserved by 1:1 v/v dilution with methanol are analyzed directly following centrifugation. Sediment samples undergo accelerated solvent extraction in aqueous methanol prior to analysis. Large volume (100 μL) sample injection and multiple reaction monitoring of a subset of the most abundant POEA homologs provide limits of quantification of 0.5 and 2.9 ng mL(-1) for MON 0818 in fresh water and sea water, respectively, and 2.5 ng g(-1) for total MON 0818 in lake sediment. Average recoveries of 93 and 75% were achieved for samples of water and sediment, respectively spiked with known amounts of MON 0818. Precision and accuracy for the analysis of water and sediment samples were within 10 and 16%, respectively based upon replicate analyses of calibration standards and representative samples. Results demonstrate the utility of the method for quantifying undegraded MON 0818 in water and sediment, although a more comprehensive method may be needed to identify and determine other POEA mixtures and degradation profiles that might occur in the environment.

  18. A novel approach to rapidly explore analytical markers for quality control of Radix Salviae Miltiorrhizae extract granules by robust principal component analysis with ultra-high performance liquid chromatography-ultraviolet-quadrupole time-of-flight mass spectrometry.

    Science.gov (United States)

    Song, Jing-Zheng; Li, Song-Lin; Zhou, Yan; Qiao, Chun-Feng; Chen, Shi-Lin; Xu, Hong-Xi

    2010-11-02

    In a well-controlled experiment, outliers discriminated by robust principal component analysis (RPCA) represent contents in samples which are of particular quality distinguishable from the rest of the others, therefore chemical constituents in a natural product causing discrimination between outliers and the majority of samples could be considered as analytical markers for quality control. Based on this strategy, a novel approach for rapidly exploring characteristic analytical markers was proposed for the quality control of extract granules of Radix Salviae Miltiorrhizae (EGRSM). In this study, large sizes of samples were analyzed via high-throughput ultra-high performance liquid chromatography-ultraviolet-quadrupole time-of-flight mass spectrometry (UHPLC-UV-Q-Tof MS). RPCA was first performed on the three groups of samples: RSM (the raw material), the in-house prepared aqueous extract of Radix Salviae Miltiorrhizae (AERSM) and commercial product of EGRSM, to determine the variation of specific constituents between raw material and the final products as well as the effect of manufacturing process on the overall quality. Then RPCA was performed on the commercial products of EGRSM to explore the applicability of identified characteristic markers for the quality control of EGRSM. Candidate markers were extracted by RPCA, and their molecular formulae were determined by high resolution electrospray ionization-mass spectrometric (ESI-MS) analysis. The suitability of identified markers was then evaluated by determining the relationship between quantities of the identified markers with their antioxidant activities biologically, and further confirmed in a variety of samples. In conclusion, the combination of RPCA with UHPLC-UV-Q-Tof MS is a reliable means to identify chemical markers for evaluating quality of herbal medicines.

  19. Trends in High Performance Liquid Chromatography for Cultural Heritage.

    Science.gov (United States)

    Degano, Ilaria; La Nasa, Jacopo

    2016-04-01

    The separation, detection and quantitation of specific species contained in a sample in the field of Cultural Heritage requires selective, sensitive and reliable methods. Procedures based on liquid chromatography fulfil these requirements and offer a wide range of applicability in terms of analyte types and concentration range. The main applications of High Performance Liquid Chromatography in this field are related to the separation and detection of dyestuffs in archaeological materials and paint samples by reversed-phase liquid chromatography with suitable detectors. The relevant literature will be revised, with particular attention to sample treatment strategies and future developments. Reversed phase chromatography has also recently gained increasing importance in the analysis of lipid binders and lipid materials in archaeological residues: the main advantages and disadvantages of the new approaches will be discussed. Finally, the main applications of ion chromatography and size exclusion chromatography in the field of Cultural Heritage will be revised in this chapter.

  20. 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 generator

  1. Thermal expansion pump for capillary high-performance liquid chromatography.

    Science.gov (United States)

    Tao, Qian; Wu, Qian; Zhang, Xiangmin

    2010-02-01

    A thermal expansion pump (TEP) based on a principle of liquid thermal expansion for capillary high-performance liquid chromatography has been developed. The novel pump is capable of generating a continuous flow at high pressure for constant and stable delivery of binary solvents from nanoliters to microliters per minute without splitting. Theoretical equations for controlling fluidic output of this pump have been established and validated by a series of experiments. Factors affecting flow rate, such as density discrepancy, liquid compressibility, and mass loss in output, were taken into account. An assembly of the pump system employing two groups of thermal expansion pumps (TEPs) working in turns were fabricated, and a controlling strategy for the pump system to maintain a continuous delivery without pressure fluctuation even at switching points was also developed. Both isocratic and gradients of binary solvent delivery by the TEPs were performed. Reproducibility and standard deviation at different flow rates were determined. A capillary high-performance liquid chromatography (micro-HPLC) system consisting of the TEPs, an injection valve, a homemade packed capillary column (20 cm x 100 microm i.d. with 5 microm C18), and a laser-induced fluorescence detector was set up, and sample separations were carried out. Results of RSD = 4% for flow and RSD = 2% for retention times at 500 nL/min were achieved. Such a pump system has almost no moving parts except for the solvent switches. Its overall costs of manufacture and running are very low. It is proven that the TEPs system has great potential and competitive capabilities in capillary liquid chromatography.

  2. Gas and high-performance liquid chromatography of phenols

    Energy Technology Data Exchange (ETDEWEB)

    Tesarova, E.; Pacakova, V.

    1983-05-01

    Gas (GC) and high-performance liquid chromatographic (HPLC) methods in the analysis of phenols are reviewed. Among the great number of phenolic compounds analyzed, alkylphenols, chlorophenols, dihydroxy-and trihydroxy-benzenes and biphenols are chiefly considered. The advantages and drawbacks of the methods are discussed. Relationships between the structural characteristics of phenols, the stationary phase structure, the mobile phase composition and the retention data are treated. Typical examples of the conditions for GC and HPLC analysis are summarized in tables. 276 references

  3. Rapid diagnosis of trisomy 21 syndrome by denaturing high-performance liquid chromatography%变性高效液相色谱法快速诊断21三体综合征

    Institute of Scientific and Technical Information of China (English)

    黄肖利; 梅端容; 黄毅; 伍严安; 陈豪

    2014-01-01

    Objective To detect the double DNA of trisomy 21 syndrome patients by denaturing high-performance liquid chro-matography(DHPLC) in order to rapidly diagnose trisomy 21 syndrome .Methods To amplify DNA fragments of three short tan-dem repeats of D21S111 ,D21S1411 and D21S1412 using corresponding primers .Then DHPLC was introduced to analyze the DNA fragments in the temperature of 50 ℃ .Results DHPLC elution profiles of D21S111 ,D21S1411 and D21S1412 of normal control showed one peak or two peaks with the same altitude .However DHPLC elution profiles of 21 trisomy syndrome patients showed two peaks of different altitudes ,which one′s altitude was twice than another .Conclusion DHPLC is a sensitive ,convenient ,auto-matic and highly-efficient method to diagnose trisomy 21 syndrome and can be widely used in the clinic diagnosis .%目的:用变性高效液相色谱法(DHPLC)分析三体综合征患者的双链DNA ,快速产前诊断21三体综合征。方法根据所设计的引物对21号染色体D21S11、D21S1411和D21S1412共3个短串联重复序列(STR)位点进行 PCR扩增,然后在50℃柱温条件下用DHPLC对PCR扩增产物进行检测和分析。结果健康对照者D21S11、D21S1411和D21S1412的DHPLC峰形呈现一个或两个高低相同的波峰,21三体综合征患者的DHPLC峰形呈现两个高低不同的波峰,且其中一个波峰高度接近为另一个波峰的2倍。结论 DHPLC具有灵敏度高、特异性强、简便快捷等,适合广泛应用于21三体综合征的快速诊断。

  4. Rapid determination of parabens in seafood sauces by high-performance liquid chromatography: A practical comparison of core-shell particles and sub-2 μm fully porous particles.

    Science.gov (United States)

    Ye, Jing; Cao, Xiaoji; Cheng, Zhuo; Qin, Ye; Lu, Yanbin

    2015-12-01

    In this work, the chromatographic performance of superficially porous particles (Halo core-shell C18 column, 50 mm × 2.1 mm, 2.7 μm) was compared with that of sub-2 μm fully porous particles (Acquity BEH C18 , 50 mm × 2.1 mm, 1.7 μm). Four parabens, methylparaben, ethylparaben, propylparaben, and butylparaben, were used as representative compounds for calculating the plate heights in a wide flow rate range and analyzed on the basis of the Van Deemter and Knox equations. Theoretical Poppe plots were constructed for each column to compare their kinetic performance. Both phases gave similar minimum plate heights when using nonreduced coordinates. Meanwhile, the flat C-term of the core-shell column provided the possibilities for applying high flow rates without significant loss in efficiency. The low backpressure of core-shell particles allowed this kind of column, especially compatible with conventional high-performance liquid chromatography systems. Based on these factors, a simple high-performance liquid chromatography method was established and validated for the determination of parabens in various seafood sauces using the Halo core-shell C18 column for separation. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  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. [Determination of amygdalin in hawthorn by high performance liquid chromatography].

    Science.gov (United States)

    Lü, Weifeng; Ding, Mingyu

    2005-09-01

    A suitable method for extraction of amygdalin from hawthorn has been established. At first, the lipophilic components were removed with petroleum ether by ultrasonic extraction. The amygdalin was then extracted by methanol in a Soxhlet's apparatus. For quantitation, a high performance liquid chromatographic method was developed by using a reversed-phase C18 column, mobile phase of methanol-water (15:85, v/v) and a detection wavelengh of 215 nm. It can be concluded that the content of amygdalin is higher in the seeds than that in the hawthorn powder without the seeds and the yield of amygdalin is higher in the hawthorn pieces than that in the hawthorn powder.

  7. Rapid quantification of tiopronin in human plasma by high-performance liquid chromatography-mass spectrometry%HPLC-MS法快速定量分析人血浆中硫普罗宁浓度

    Institute of Scientific and Technical Information of China (English)

    刘娜; 张蓓蓓; 张尊建; 田媛; 许风国; 陈洓

    2008-01-01

    Aim:To develop a rapid high-performance liquid chromatography-mass spectrometry (HPLC-MS) method for the quantification of tiopronin in human plasma.Methods:Cysteine was chosen as antioxidant and firstly added into the whole blood firstly.After adding mycophenolic acid as internal standard (IS) and 1 mol/L HCl into the plasma,the samples were extracted with acetic ether and then determined by HPLC-MS.The chromatographic separation was performed on a Shim-pack VP-ODS C18 column (250 mm×2.0 mm,5 μm) using methanol-0.1% acetic acid (70∶30) as mobile phase with a flow rate of 0.2 mL/min.Results:The assay exhibited a linear range of tiopronin between 30~4 000 ng/mL.The precisions for intra- and inter-batch were all within 8.5%.The extraction recoveries were more than 70%.The total HPLC-MS analysis time was within 7.5 min per a run.The fully validated method was successfully applied to quantify tiopronin in human plasma for a bioavailability study.Conclusion:The assay proved to be accurate,sensitive,selective and convenient.The fully validated method can be applied to study the pharmacokinetics and bioavailability of tiopronin and tiopronin formulations in human.%目的:建立快速测定人血浆中硫普罗宁浓度的HPLC-MS联用分析方法.方法:在全血中加入抗氧剂半胱氨酸后,于分得的血浆中加入霉酚酸为内标,酸化后经乙酸乙酯提取,进行HPLC-MS测定.色谱柱为Shim-pack VP-ODS C18 column (250 mm×2.0 mm,5 μm),流动相为甲醇-0.1 %乙酸(70:30),流速为0.2 mL/min.结果:方法的线性范围为30~4 000 ng/mL,提取回收率大于70%,批间和批内精密度均小于8.5%,单样品分析时间7.5 min.并将所建立的方法成功地应用于硫普罗宁片生物等效性研究.结论:本方法操作简单、快速,灵敏度、准确度、精密度和定量分析线性关系均良好,符合生物样品测定要求.

  8. Rapid profiling and identification of triterpenoid saponins in crude extracts from Albizia julibrissin Durazz. by ultra high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry.

    Science.gov (United States)

    Han, Lifeng; Pan, Guixiang; Wang, Yuefei; Song, Xinbo; Gao, Xiumei; Ma, Baiping; Kang, Liping

    2011-07-15

    Ultra high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UHPLC/ESI-Q-TOF-MS/MS) was applied to separate and identify triterpenoid saponins in crude extract from the stem bark of Albizia julibrissin Durazz. The molecular weights were determined by comparing quasi-molecular ions [M+NH(4)](+) in positive mode and [M-H](-) and [M-2H](2-) ions in negative mode. The MS/MS spectra of the [M-H](-) ions for saponins provided a wealth of structural information related to aglycone skeletons, sugar types and linked sequence. On the basis of the fragmentation behavior of known saponins isolated before, saponins from this plant were identified, even though references were not available. As a result, a total of twenty-eight saponins in the crude extract were identified, which all had a common basic skeleton of the triterpene oleanolic acid and eight of them were new compounds.

  9. Analysis of Free Fatty Acids on the Fingertips by High Performance Liquid Chromatography.

    Science.gov (United States)

    1978-12-20

    This investigation studied the efficiency of high performance liquid chromatography in the determination of free fatty acids present on the...utilized to eliminate the microbial contamination. The high performance liquid chromatography provided excellent separation of skin fatty acids for

  10. Rapid Prototyping of High Performance Signal Processing Applications

    Science.gov (United States)

    Sane, Nimish

    Advances in embedded systems for digital signal processing (DSP) are enabling many scientific projects and commercial applications. At the same time, these applications are key to driving advances in many important kinds of computing platforms. In this region of high performance DSP, rapid prototyping is critical for faster time-to-market (e.g., in the wireless communications industry) or time-to-science (e.g., in radio astronomy). DSP system architectures have evolved from being based on application specific integrated circuits (ASICs) to incorporate reconfigurable off-the-shelf field programmable gate arrays (FPGAs), the latest multiprocessors such as graphics processing units (GPUs), or heterogeneous combinations of such devices. We, thus, have a vast design space to explore based on performance trade-offs, and expanded by the multitude of possibilities for target platforms. In order to allow systematic design space exploration, and develop scalable and portable prototypes, model based design tools are increasingly used in design and implementation of embedded systems. These tools allow scalable high-level representations, model based semantics for analysis and optimization, and portable implementations that can be verified at higher levels of abstractions and targeted toward multiple platforms for implementation. The designer can experiment using such tools at an early stage in the design cycle, and employ the latest hardware at later stages. In this thesis, we have focused on dataflow-based approaches for rapid DSP system prototyping. This thesis contributes to various aspects of dataflow-based design flows and tools as follows: 1. We have introduced the concept of topological patterns, which exploits commonly found repetitive patterns in DSP algorithms to allow scalable, concise, and parameterizable representations of large scale dataflow graphs in high-level languages. We have shown how an underlying design tool can systematically exploit a high

  11. Progress on high-performance rapid prototype aluminum mirrors

    Science.gov (United States)

    Woodard, Kenneth S.; Myrick, Bruce H.

    2017-05-01

    Near net shape parts can be produced using some very old processes (investment casting) and the relatively new direct metal laser sintering (DMLS) process. These processes have significant advantages for complex blank lightweighting and costs but are not inherently suited for producing high performance mirrors. The DMLS process can provide extremely complex lightweight structures but the high residual stresses left in the material results in unstable mirror figure retention. Although not to the extreme intricacy of DMLS, investment casting can also provide complex lightweight structures at considerably lower costs than DMLS and even conventional wrought mirror blanks but the less than 100% density for casting (and also DMLS) limits finishing quality. This paper will cover the progress that has been made to make both the DMLS and investment casting processes into viable near net shape blank options for high performance aluminum mirrors. Finish and figure results will be presented to show performance commensurate with existing conventional processes.

  12. Determination of a Jet Fuel Metal Deactivator by High Performance Liquid Chromatography

    Science.gov (United States)

    1983-06-01

    HIGH PERFORMANCE LIQUID CHROMATOGRAPHY Paul C. Hayes, Jr. Fuels Branch...SUPPLEMENTARY NOTES 19. KEY WORDS (Continue on reverse side if necessary and identify by block number) High Performance Liquid Chromatography absorbance...SYMBOL HPLC High Performance Liquid Chromatography P-4 jet propulsion fuel, wide-boiling range, conforming to MIL-T-5624L MDA metal deactivator,

  13. Determination of Stabiliser Contents in Advanced Gun Propellants by Reverse Phase High Performance Liquid Chromatography

    Science.gov (United States)

    1994-03-01

    HIGH PERFORMANCE LIQUID CHROMATOGRAPHY N"m A.R. TURNER AND A. WHITE...TO biEPROOU.; AND SELL THIS REPORT Determination of Stabiliser Contents in Advanced Gun Propellants by Reverse Phase High Performance Liquid Chromatography A.R...8217/......... .. Availability Cooes Dist Avaiardlo A-i Determination of Stabiliser Contents in Advanced Gun Propellants by Reverse Phase High Performance Liquid Chromatography

  14. High Performance Liquid Chromatographic Method for Determination of Dipyridamole in Human Plasma

    Directory of Open Access Journals (Sweden)

    DAVOOD BEIGI BAND ARAB ADI

    1999-08-01

    Full Text Available A simple, rapid and specific high-performance liquid chromatographic procedure is reported for"nquantitative determination of dipyridamole in human -plasma. The assay uses a reversed-phase"nhigh-performance liquid chromatographic (HPLC and UV detection at 280nm and has a limit"nof detection of approximately 5ng/mL. The mobile phase consists of MeOH-H20 (60:40"nadjusted to pH 3.3. Dipyridamole was extracted from plasma by back-extraction procedure, with"npropranolol as the internal standard. The reproducibility of the method is satisfactory

  15. Determination of saffron quality by high-performance liquid chromatography.

    Science.gov (United States)

    Valle García-Rodríguez, M; Serrano-Díaz, Jéssica; Tarantilis, Petros A; López-Córcoles, Horacio; Carmona, Manuel; Alonso, Gonzalo L

    2014-08-13

    The aim of this work was to propose a high-performance liquid chromatography with diode array detection (HPLC-DAD) method for determining the three main compounds responsible for determining the quality of saffron (crocetin esters, picrocrocin, and safranal) by preparing an aqueous extract according to the ISO 3632 standard to solve the difficulty that this standard has for aroma and taste determination by ultraviolet-visible spectroscopy. Toward this aim, laboratory-isolated picrocrocin, a safranal standard with a purity of ≥ 88%, trans-crocetin di(β-D-gentiobiosyl) ester (trans-4-GG) and trans-crocetin (β-D-glucosyl)-(β-D-gentiobiosyl) ester (trans-3-Gg) standards, both with a purity of ≥ 99%, and 50 different saffron spice samples from Italy, Iran, Greece, and Spain were used in the intralaboratory validation of the HPLC method. The analytical method proposed was adequate in terms of linearity, selectivity, sensitivity, and accuracy for determining the three foremost parameters that define the quality of saffron using only a saffron solution prepared according to the ISO 3632 standard.

  16. Gradient chromatofocusing high-performance liquid chromatography. I. Practical aspects.

    Science.gov (United States)

    Liu, Y; Anderson, D J

    1997-02-21

    In this work, a versatile method for generating linear pH gradients using weak anion-exchange HPLC has been developed, which is termed gradient chromatofocusing high-performance liquid chromatography. This method utilizes a linear external pH gradient generated in the mobile phase entering the column (inlet pH gradient), superimposed on an internally-generated pH gradient within the column (column pH gradient), which results from the buffering action of the ion exchanger on the mobile phase and vice versa. The method shows significant advantages over conventional chromatofocusing, including: decreased expense due to the use of common buffer components, ease of adjusting the slope of the pH gradient produced at the outlet of the column (outlet pH gradient) through the manipulation of the inlet pH gradient and the ability of using high concentration buffers in the mobile phase. Chromatography of fibrinogen degradation products was done using gradient chromatofocusing. Bandwidths comparable to conventional chromatofocusing were obtained in the separation of fibrinogen degradation products.

  17. Gradient chromatofocusing high-performance liquid chromatography. II. Theoretical aspects.

    Science.gov (United States)

    Liu, Y; Anderson, D J

    1997-02-21

    This article is Part II of a series describing a newly-developed gradient chromatofocusing high-performance liquid chromatography (HPLC) technique. Theoretical aspects of the technique are discussed. In gradient chromatofocusing, the column pH gradient with respect to column distance can be varied without necessarily affecting the outlet pH gradient with respect to time. Factors influencing the value of the slope of the column pH gradient are identified through derived equations and a computer simulation model. A newly-identified parameter is introduced, column travel time, which can be uniquely varied in gradient chromatofocusing. Experiments show increased conversion of fibrinogen to denatured forms with increased column travel time. Another unique aspect of gradient chromatofocusing is that the mobile phase buffer concentration can be manipulated without necessarily affecting the outlet pH gradient slope, giving the technique expanded versatility for optimizing the separation. In the present work, the pIapparent for fibrinogen is found to increase with increased mobile phase buffer concentration.

  18. Rapid and specific high-performance liquid chromatography for the in vitro quantification of D-Lys6-GnRH in a microemulsion-type formulation in the presence of peptide oxidation products.

    Science.gov (United States)

    Kafka, Alexandra P; Rades, Thomas; McDowell, Arlene

    2010-02-01

    A high-performance liquid chromatography (HPLC) method for assay of d-Lys(6)-GnRH contained in a microemulsion-type formulation is described. The peptide is extracted from the microemulsion matrix and quantified using a two-step gradient method. Separation from microemulsion compounds and potential peptide oxidation products was achieved on a Jupiter C(18) column at 40 degrees C, using a gradient of 10-35% CH(3)CN for peptide elution. The correlation of peak intensity measured at 220 nm and peptide concentration was linear over the range 2.5-60 microg/mL with a correlation coefficient of 0.9997 and a y-intercept not significantly different from zero (p > 0.05). Intraday and interday variability of the assay was less than 5% for multiple injections of samples containing 7.5, 30 and 60 microg/mL. The lower limit of quantitation was calculated to be 0.38 microg/mL, and the lower limit of detection was 0.13 microg/mL. The assay was applied to samples that were stressed under physiological conditions (37 degrees C, pH 7.4) over 4 days. Three degradation peaks were well resolved from the parent peptide, demonstrating the selectivity of the assay. Off-line MALDI TOF mass spectrometry was applied to identify these degradation species as oxidation products of the peptide.

  19. Development of an ultra-high-performance liquid chromatography coupled to high-resolution quadrupole-Orbitrap mass spectrometry method for the rapid detection and confirmation of illegal adulterated sedative-hypnotics in dietary supplements.

    Science.gov (United States)

    Jiang, Shuyin; Tan, Huijie; Guo, Changchuan; Gong, Liping; Shi, Feng

    2015-01-01

    A novel method using ultra-high performance liquid chromatography coupled to hybrid quadrupole-Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap) was developed and validated for the simultaneous screening, identification and quantification of sedative-hypnotics in dietary supplements. Chromatographic conditions were optimised and a full data-dependent MS(2) scan (MS/dd-MS(2)) in positive and negative ion mode was used. A single injection was sufficient to perform the simultaneous screening and identification/quantification of samples. The response showed a good linear relationship with analyte concentrations over wide ranges (e.g., 1.0-1000 ng g(-1) for diazepam) with all the determination coefficients (r(2)) > 0.9985. The method was validated, obtaining accuracy (intra- and inter-day) in the range of 94.5-105.3% and precision (intra- and inter-day) in the range of 0.4-8.9%, respectively. The detection limits (LODs) were in the range of 0.3-1.0 ng g(-1) for different analytes. Recoveries were performed and ranged from 74.1% to 90.2%, while all matrix effects were over the range of 85.4-93.6%. Finally, this method was used to detect sedative-hypnotics in commercial dietary supplements. Of a total of 45 batches of dietary supplements, only three batches were found to be positive samples with concentrations of diazepam, clonazepam and alprazolam at high levels (≥ 8.22 mg g(-1)).

  20. Determination of acaricides in honey by high-performance liquid chromatography with photodiode array detection.

    Science.gov (United States)

    Martel, Anne-Claire; Zeggane, Sarah

    2002-04-19

    Rapid analytical methods are described to control quality of honeys, concerning residues of acaricides applied in hives to prevent Varroa jacobsoni infestation. A liquid-liquid extraction with hexane-propanol-2-ammonia (60 ml:30 ml:0.28%) was used for the simultaneous analysis of coumaphos, bromopropylate, amitraz and fluvalinate. For thymol, one clean up on a solid-phase extraction C18 (500 mg, 6 ml) column was performed; for rotenone, a liquid extraction with dichloromethane was realised. Quantitative recoveries obtained with honey were satisfactory and were superior to 80%. All acaricides are identified by reversed-phase high-performance liquid chromatography with photodiode array detection. Quantification limits obtained were below maximal residue limits when these exist.

  1. Determination of Polysaccharide in Radix pseudostellariae Extract by Size-Exclusion High-Performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    A simple size-exclusion high-performance liquid chromatographic method was developed for rapid molecular mass screening to determine the average molecular mass, polydispersity, and quantity of Taizishen polysaccharides extract. The screening used a TOSOH TSK-GEL G3000SW high-performance liquid chromatographic column with 0.1 mol/L NaNO3 buffer as the mobile phase and refractive index detector. The molecular mass calibration curve was linear for polysaccharide standards from 10 to 100 kDa with a correlation coefficient of 0.9991. The method can be used to analyze the quantity, average molecular mass,and polydispersity of polysaccharides. In addition, the screening method is suitable for quality control of polysaccharide preparations in Chinese medicines and pharmaceutical products.

  2. Determination of fenbendazole, praziquantel and pyrantel pamoate in dog plasma by high-performance liquid chromatography.

    Science.gov (United States)

    Morovján, G; Csokán, P; Makranszki, L; Abdellah-Nagy, E A; Tóth, K

    1998-02-27

    Simple and rapid high-performance liquid chromatographic methods were developed for the determination of fenbendazole, praziquantel and pyrantel pamoate in dog plasma. The combination of these drugs is the most powerful treatment against most types of worms. Blood plasma samples obtained in a pharmacokinetic trial were prepared using solid-phase extraction. Fenbendazole and praziquantel were analyzed simultaneously by reversed-phase high-performance liquid chromatography on an octadecyl-modified silica stationary phase employing acetonitrile-phosphate buffer (pH 3.0) eluent and ultraviolet detection at 220 nm. Pyrantel was analyzed separately on a base-deactivated reversed-phase column using methanol-tetrahydrofuran-ammonium acetate buffer (pH 4.6) eluent and ultraviolet detection at 317 nm. Average recoveries for fenbendazole, praziquantel and pyrantel pamoate were 76.8, 93.4 and 90.5%, respectively. Limits of quantitation were in the range of 15-25 ng/ml plasma.

  3. Rapid Euler CFD for High-Performance Aircraft Design

    Science.gov (United States)

    Charlton, Eric F.

    2004-01-01

    The goal here was to present one approach to rapid CFD for S&C using an unstructured inviscid method, in order to eventually assess S&C properties as early in the design process as possible. Specific results are presented regarding time, accuracy (as compared to a baseline wind tunnel database) and simplicity for the user. For COMSAC, it s more important to talk about the "specifications" required by Advanced Design and S&C, as well as how the CFD results can be combined for envelope evaluation.

  4. Analysis of ganciclovir and its related substances using high performance liquid chromatography and liquid chromatography-mass spectrometry methods

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Objective High performance liquid chromatography(HPLC)and liquid chromatography-mass spectrometry(LC/MS)methods were developed for the determination of ganciclovir and its related substances.Methods A Hypersil ODS2 column(4.6 mm×250 mm,5 μm)was used with a mobile phase of 0.02 M potassium dihydrogen phosphate buffer(pH 6.0)-methanol(92∶8)at a flow rate of 1.0 mL/min,and UV detector set at 254 nm was used for monitoring the eluents.Results The method was simple,rapid,selective and capable of separating all r...

  5. Dispersive Liquid-Liquid Microextraction and determination of Platinum(IV by High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Ali Mazloomifar

    2015-09-01

    Full Text Available A simple, rapid, and efficient procedure, dispersive liquid-liquid microextraction (DLLME, has been developed for the extraction and preconcentration of platinum (IV in environmental water samples. The factors relevant to the microextraction efficiency, such as the kind and volume of extraction and dispersive solvent, the extraction time, the pH in aqueous, and the salt effect, were optimized. Under the optimum conditions , the enrichment factor of this method for platinum was reached at 119. The detection limit for platinum was 0.3 ng mL-1 , and the relative standard deviation (RSD was 0.42% (n = 10 , C = 10 ng mL-1. The method was successfully applied to the determination of trace amounts of platinum in environmental water samples.

  6. 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)

  7. A rapid method for the determination of perfluoroalkyl substances including structural isomers of perfluorooctane sulfonic acid in human serum using 96-well plates and column-switching ultra-high performance liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Salihovic, Samira; Kärrman, Anna; Lindström, Gunilla; Lind, P Monica; Lind, Lars; van Bavel, Bert

    2013-08-30

    To facilitate high-throughput analysis suitable for large epidemiological studies we developed an automated column-switching ultra-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for determination of perfluorocarboxylic acids (PFCAs; C5, C6, C7, C8, C9, C10, C11, C12, and C13), perfluoroalkyl sulfonic acids (PFSAs; C4, C6, C8, and C10), perfluorooctane sulfonamide (PFOSA), and five groups of structural perfluorooctane sulfonic acid (PFOS) isomers in human serum or plasma. The analytical procedure involves rapid protein precipitation using 96-well plates followed by an automated sample clean-up using an on-line trap column removing many potentially interfering sample components while through the mobile phase gradient the target analytes are eluted onto the analytical column for further separation and subsequent mass detection. The method was linear (R(2)method detection limits ranging between 0.01 and 0.17ngmL(-1) depending on the analyte. The developed method was precise, with repeatability (n=7) and reproducibility (n=103) coefficients of variation between 2% and 20% for most compounds including PFOS (2% and 8%) and its structural isomers (2-6% and 4-8%). The method was in conformity with a standard reference material. The column-switching HPLC-MS/MS method has been successfully applied for the determination of perfluoroalkyl substances including structural PFOS isomers in human plasma from an epidemiological study.

  8. Rapid determination of 30 bioactive constituents in XueBiJing injection using ultra high performance liquid chromatography-high resolution hybrid quadrupole-orbitrap mass spectrometry coupled with principal component analysis.

    Science.gov (United States)

    Zuo, Lihua; Sun, Zhi; Hu, Yurong; Sun, Ya; Xue, Wenhua; Zhou, Lin; Zhang, Jun; Bao, Xiaoyue; Zhu, Zhenfeng; Suo, Guanglu; Zhang, Xiaojian

    2017-04-15

    Xuebijing injection (XBJ) is a traditional Chinese herbal prescription widely used in the treatment of sepsis. Extensive chemical studies revealed that XBJ injection contains amino acids, phenolic acids, flavonoid glycosides, terpeneglycosides and phthalides. In this study, the applicability of ultra high performance liquid chromatography coupled with high resolution hybrid quadruple-orbitrap mass spectrometry (UHPLC-Q-Orbitrap MS) for the simultaneous quantitative analysis of 30 bioactive constituents in XueBiJing injection (XBJ) was investigated. The mass spectrometer was operated in full MS scan mode. The use of 70,000FWHM mass resolution and narrow mass windows (5ppm) could effectively improve the selectivity of the method. Separation was achieved on a Waters ACQUITY UPLC(®) HSS C18 column (2.1mm×100mm, 1.8μm) with a gradient mobile phase consisting of acetonitrile-water (containing 10mM ammonium acetate) at a flow rate of 0.2mL/min. Satisfactory linearity was achieved within wide linear range and all correlation coefficients (r) of analytes were more than 0.9996. The limits of detection (LODs) were in the range of 0.1180-27.82ng/mL for different analytes. The relative standard deviations (RSDs) of inter- and intra-day precisions were less than 3.0% and the recoveries of the assay were in the range of 98.5%-101.5%. The validated method was successfully applied for simultaneous determination of 30 bioactive compounds in XueBiJing injection from 10 batches samples by UHPLC-Q-Orbitrap MS within 10min. Moreover, the results were evaluated principal component analysis and two compounds might be the most important chemical markers for chemical quality control of XBJ injection. The novel Q-Orbitrap mass spectrometry has been proved to be a very promising and powerful tool for routine screening of bioactive compounds in traditional Chinese medicine injection, ensuring drug safety and public health.

  9. Rapid Determination of Capsaicinoids in Hotpot Seasoning by High-Performance Liquid Chromatography%高效液相色谱法测定火锅底料中辣椒素类物质

    Institute of Scientific and Technical Information of China (English)

    尼海峰; 但晓容; 李栋钢

    2014-01-01

    A rapid method of high-pressure liquid chromatography (HPLC)is developed to determine the pungency components (capsaicin and dihydrocapsaicin)in hotpot seasoning.The column is RP-C1 8 (4.6 mm×250 mm,5 μm).The mobile phase is mixture of methanol and water (65 ∶35 )with an isocratic flow of 1 mL/min.The temperature of column is 30 ℃,the injection volume is 10 μL,and the UV wavelength is 280 nm.Ultrasonic assisted extraction is used to extract the capsaicinoids in hotpot seasoning.It shows that the content of capsaicin and dihydrocapsaicin is 0.256 ~0.81 6 g/kg, Scoville index is 3948~12583 SHU.%为了测定火锅底料中的辣味物质(辣椒素及二氢辣椒素),建立了高效液相色谱快速检测方法:色谱柱为 C18 InertSustain (4.6 mm ×250 mm,5μm),流动相为甲醇和水(V/V,65∶35),流速1.0 mL/min,柱温30℃,进样量10μL,检测波长280 nm。辣椒素类物质的提取采用超声波辅助提取。结果表明:火锅底料中的辣椒素类物质含量在0.256~0.816 g/kg 之间,斯科维尔系数在3948~12583 SHU。

  10. Determination of Selected Colored Smokes on Glass Fiber Discs by High Performance Liquid Chromatography (HPLC)

    Science.gov (United States)

    1991-05-01

    High Performance Liquid Chromatography (HPLC) 12. PERSONAL AUTHOR(S) F F_ n.ipl’prifl. Alan R...GROUP SUB-GROUP High Performance Liquid Chromatography (HPLC), Analytical IMethod, 1,4-diamino-2,3-dihydroanthraquinone, 2-(2 - _ quinolinyl)-1,3...weights, low vapor pressures and low thermal stability. High performance liquid chromatography (HPLC) appears to be the analytical method of choice

  11. Quantitative high-performance liquid chromatographic determination of delta 4-3-ketosteroids in adrenocortical extracts.

    Science.gov (United States)

    Ballerini, R; Chinol, M; Ghelardoni, M

    1980-05-30

    A high-performance liquid chromatographic method is described for the determination of seven steroids in adrenocortical extracts showing a delta 4-3-ketonic conjugated system. The seven steroids (cortisol, cortisone, 11-dehydrocorticosterone, corticosterone, 11-deoxycortisol, aldosterone and 11-deoxycorticosterone) were separated with a chloroform-methanol gradient on a 5-micron silica column and with a water-acetonitrile gradient on a 10-micron RP-8 column. Effluents were monitored by UV absorption at 242 nm. Quantitative analysis was performed by comparing peak areas, which are proportional to the amounts of the individual substances (external standard method). The method is rapid, sensitive, easy to perform and reproducible.

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

    Directory of Open Access Journals (Sweden)

    Mehdi Maham

    2014-09-01

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

  13. Rapid determination of monoamine neurotransmitters and their metabolites by high performance liquid chromatography%高效液相色谱法快速检测单胺类神经递质及其代谢产物

    Institute of Scientific and Technical Information of China (English)

    伦立民; 车琳杰; 李世伟

    2006-01-01

    BACKGROUND: The determination of monoamine neurotransmitters and their metabolites is important in the experimental.research of Parkinson disease (PD) and experimental diagnosis of phaochromocytoma.OBJECTIVE: To develop a high performance liquid chromatography (HPLC) method for simultaneous and rapid determination of monoamine neurotransmitters and their metabolites which include epinephrine(E), norepinephrine(NE), dopamine(DA), 5'-hydroxytryptamine(5'-HT), 5-hydroxy3-indoleacetic acid(5-HIAA), homovanillic acid(HVA) and 3,4-dihydroxyphenylacetic acid(DOPAC).DESIGN: Case control observation and a randomized controlled animal experiment.SETTING: Department of Clinical Laboratory, Affiliated Hospital of Medical College, Qingdao University; Haiyang Branch, Affiliated Hospital of Medical College, Qingdao University.PARTICIPANTS: This experiment was carried out at Staff Room of Physiology, edical College, Qingdao University and Department of Clinical Laboratory, Affiliated Hospital of Medical College of Qingdao University from October to December 2003. ①Two patients with phaeochromocytoma were all confirmed by CT, MRI or pathologic section as well as physical sign. Twelve health volunteers were all university students and were excluded to suffer hypertension, endocrine system diseases and other diseases. ② Total 12 healthy female Wister rats weighed between 150 g to 180 g were chosen and randomly divided into control group and PD group with 6 rats in each.METHODS: The standard chromatograms of the standard solution of mixtures of seven substances (E, NE, DA, 5'-HT, 5-HIAA, HVA and DOPAC)were prepared and the precisions and recovery rates of the same samples were determined respectively. Urine samples of 24 hours obtained from 12 healthy volunteers and 2 patients with phaeochromocytoma were analyzed respectively by HPLC. The unilateral forebrain bundles in PD group were destroyed with 6-hydroxydopamine (6-OHDA) for monitoring release characteristics of monoamine

  14. 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)

  15. [Qualitative and quantitative analysis of the amino acids in Rhizoma Arisaematis by ultra high performance liquid chromatography-tandem mass spectrometry and high performance liquid chromatography].

    Science.gov (United States)

    Wang, Xing; Chi, Yumei; Kang, An

    2014-12-01

    A method for the identification and determination of the polar amino components without ultraviolet activity in traditional Chinese medicines was developed. With Rhizoma Arisaematis as the object of this study, using pre-column derivatization with phenyl isothiocyanate (PITC) as the derivatization reagent, compounds were separated and identified on a C18 column (100 mm x 2.1 mm, 3.5 µm) by ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). A total of 20 components, including 18 amino acids and 2 amine compounds were identified. Furthermore, after the optimization of the derivatization conditions, 15 amino acids were determined by high performance liquid chromatography (HPLC) on Diamonsil C18 column (250 mm x 4.6 mm, 5 µm), detected at 254 nm and gradiently eluted by acetonitrile and 0. 05 mol/L ammonium acetate-acetic acid (pH 6. 5) as the mobile phases. The results of methodological study demonstrated that the method can meet the requirements of the determination. All calibration curves expressed good linearity: Glu, Try in the range of 2-100 mg/L, Arg in the range of 6-300 mg/L, others in the range of 0. 8-40 µg/L, with the correlation coefficients ≥ 0. 999 5. The average recovery of this method was among 95%-105% and the RSD was less than 3%. The developed method was successfully applied to quantitative determination of amino compounds in 12 batches of Rhizoma Arisaematis samples. The method is simple, sensitive, accurate, and can be used for rapid identification and determination of amino components in traditional Chinese medicines.

  16. Sheath liquid effects in capillary high-performance liquid chromatography-electrospray mass spectrometry of oligonucleotides.

    Science.gov (United States)

    Huber, C G; Krajete, A

    2000-02-18

    Fused-silica capillary columns of 200 microm inner diameter were packed with micropellicular, octadecylated, 2.3 microm poly(styrene-divinylbenzene) particles and applied to the separation of oligonucleotides by ion-pair reversed-phase high-performance liquid chromatography. Oligonucleotides were eluted at 50 degrees C with gradients of 3-13% acetonitrile in 50 mM triethylammonium bicarbonate. Addition of sheath liquid to the column effluent allowed the detection of oligonucleotides by electrospray ionization mass spectrometry using full-scan data acquisition with a detectability comparable to that obtained with UV detection. The signal-to-noise ratios with different sheath liquids increased in the order isopropanolliquid was found to influence the charge state distribution of oligonucleotides longer than 20 nucleotide units whereas no significant effect was observed with shorter oligonucleotides. Organic acids and bases in the sheath liquid generally deteriorated the signal-to-noise ratios in the chromatograms and mass spectra mainly because of increased background noise. Only a few charge states were observed in the mass spectra of oligonucleotides because of charge state reduction due to the presence of carbonic acid in the eluent. With triethylammonium hydrogencarbonate as chromatographic eluent and acetonitrile as sheath liquid, very few cation adducts of oligonucleotides were observed in the mass spectra. However, the presence of small amounts of monopotassium adducts enabled the calculation of the charge state of multiply charged ions. With acetonitrile as sheath liquid, 710 amol of a 16-mer oligonucleotide were detected using selected ion monitoring data acquisition with a signal-to-noise ratio of 3:1. Finally, capillary ion-pair reversed-phase high-performance liquid chromatography-electrospray ionization mass spectrometry was

  17. High-performance liquid chromatographic assay for benzocaine and p-aminobenzoic acid including preliminary stability data.

    Science.gov (United States)

    Narang, P K; Bird, G; Crouthamel, W G

    1980-12-01

    A high-performance liquid chromatographic assay was developed that separates and quantitates benzocaine and its primary degradation product, p-aminobenzoic acid. This method is rapid, sensitive, and specific. Preliminary stability data obtained with this method demonstrate its utility for this purpose.

  18. Simultaneous determination of dapsone, monoacetyldapsone and pyrimethamine in whole blood and plasma by high-performance liquid chromatography

    DEFF Research Database (Denmark)

    Lemnge, M M; Rønn, A; Flachs, H

    1993-01-01

    A sensitive, selective and rapid reversed-phase high-performance liquid chromatographic method was developed for the simultaneous analysis of dapsone, monoacetyldapsone and pyrimethamine in human whole blood and plasma. The procedure involved extraction of the compounds and the internal standard...

  19. High-performance liquid chromatography with packed microchips

    OpenAIRE

    Ehlert, Steffen

    2011-01-01

    This work is explaining the importance of generating a densely packed chromatographic bed to achieve optimum separation efficiency in miniaturized liquid chromatography. The reduction of peak dispersion and enhancement of separation efficiency is the important problem in micro- and nano-separation that is to be solved to generate highly efficient chromatographic systems that can exploit all advantages of miniaturization. Therefore...

  20. High performance ultracapacitors with carbon nanomaterials and ionic liquids

    Science.gov (United States)

    Lu, Wen; Henry, Kent Douglas

    2012-10-09

    The present invention is directed to the use of carbon nanotubes and/or electrolyte structures in various electrochemical devices, such as ultracapacitors having an ionic liquid electrolyte. The carbon nanotubes are preferably aligned carbon nanotubes. Compared to randomly entangled carbon nanotubes, aligned carbon nanotubes can have better defined pore structures and higher specific surface areas.

  1. High performance batteries with carbon nanomaterials and ionic liquids

    Science.gov (United States)

    Lu, Wen

    2012-08-07

    The present invention is directed to lithium-ion batteries in general and more particularly to lithium-ion batteries based on aligned graphene ribbon anodes, V.sub.2O.sub.5 graphene ribbon composite cathodes, and ionic liquid electrolytes. The lithium-ion batteries have excellent performance metrics of cell voltages, energy densities, and power densities.

  2. High Performance Negative Dielectric Anisotropy Liquid Crystals for Display Applications

    Directory of Open Access Journals (Sweden)

    Xiaolong Song

    2013-09-01

    Full Text Available We review recent progress in the development of high birefringence (Δn ≥ 0.12 negative dielectric anisotropy (Δε < 0 liquid crystals (LCs for direct-view and projection displays. For mobile displays, our UCF-N2 (low viscosity, negative Δε, high Δn based homogeneous alignment fringe-field switching (called n-FFS mode exhibits superior performance to p-FFS in transmittance, single gamma curve, cell gap insensitivity, and negligible flexoelectric effect. For projection displays using a vertical alignment liquid-crystal-on-silicon (VA LCOS, our high birefringence UCF-N3 mixture enables a submillisecond gray-to-gray response time, which is essential for color sequential displays without noticeable color breakup. Our low viscosity UCF-N2 also enables multi-domain VA displays to use a thinner cell gap for achieving faster response time.

  3. High Performance Mars Liquid Cooling and Ventilation Garment Project

    Science.gov (United States)

    Terrier, Douglas; Clayton, Ronald; Whitlock, David; Conger, Bruce

    2015-01-01

    EVA space suit mobility in micro-gravity is enough of a challenge and in the gravity of Mars, improvements in mobility will enable the suited crew member to efficiently complete EVA objectives. The idea proposed is to improve thermal efficiencies of the liquid cooling and ventilation garment (LCVG) in the torso area in order to free up the arms and legs by removing the liquid tubes currently used in the ISS EVA suit in the limbs. By using shaped water tubes that greatly increase the contact area with the skin in the torso region of the body, the heat transfer efficiency can be increased to provide the entire liquid cooling requirement and increase mobility by freeing up the arms and legs. Additional potential benefits of this approach include reduced LCVG mass, enhanced evaporation cooling, increased comfort during Mars EVA tasks, and easing of the overly dry condition in the helmet associated with the Advanced Extravehicular Mobility Unit (EMU) ventilation loop currently under development.

  4. Rapid quick, easy, cheap, effective, rugged, and safe extraction with novel phospholipid cleanup: A streamlined ultra high performance liquid chromatography with ultraviolet detection approach for screening polycyclic aromatic hydrocarbons in avian blood cells and plasma.

    Science.gov (United States)

    Provatas, Anthony A; Yevdokimov, Alexander V; King, Cory A; Gatley, Emma L; Stuart, James D; Evers, David C; Perkins, Christopher R

    2015-08-01

    A streamlined method has been developed for the isolation and analysis of polycyclic aromatic hydrocarbons in avian blood cells and plasma utilizing quick, easy, cheap, effective, rugged, and safe extraction in combination with novel phospholipid cleanup technology. A variety of traditional extraction and cleanup techniques have been employed in the preparation and analysis of polycyclic aromatic hydrocarbonsin a variety of matrices; liquid-liquid partitioning, solid-phase extractions, gel permeation chromatography, and column chromatography are all effective techniques, however they are laborious and time consuming processes that require large amounts of solvent. Using quick, easy, cheap, effective, rugged, and safe extraction coupled with phospholipid cleanup, samples can be quickly screened while maintaining high throughput and sensitivity. With a liquid chromatography approach, analysis times may be kept short at 16 min while maintaining high analyte recovery. Recoveries in quality control samples ranged from 70 to 109%, with average surrogate recoveries of 80.6 ± 1.10%. The result of using a quick, easy, cheap, effective, rugged, and safe extraction approach in conjunction with phospholipid cleanup is a methodology that significantly reduces sample preparation time and solvent use while maintaining high sensitivity and reproducibility.

  5. Determination of five antiarrhythmic drugs in human plasma by dispersive liquid-liquid microextraction and high-performance liquid chromatography.

    Science.gov (United States)

    Jouyban, Abolghasem; Sorouraddin, Mohammad Hossein; Farajzadeh, Mir Ali; Somi, Mohammad Hossein; Fazeli-Bakhtiyari, Rana

    2015-03-01

    A fast and sensitive high-performance liquid chromatography (HPLC) method with ultraviolet (UV) detection was developed and validated for the simultaneous quantitation of five antiarrhythmic drugs (metoprolol, propranolol, carvedilol, diltiazem, and verapamil) in human plasma samples. It involves dispersive liquid-liquid microextraction (DLLME) of the desired drugs from 660 µL plasma and separation using isocratic elution with UV detection at 200 nm. The complete separation of all analytes was achieved within 7 min. Acetonitrile (as disperser solvent) resulting from the protein precipitation procedure was mixed with 100 µL dichloromethane (as an extraction solvent) and rapidly injected into 5 mL aqueous solution (pH 11.5) containing 1% (w/v), NaCl. After centrifugation, the sedimented phase containing enriched analytes was collected and evaporated to dryness. The residue was re-dissolved in 50 µL de-ionized water (acidified to pH 3) and injected into the HPLC system for analysis. Under the optimal conditions, the enrichment factors and extraction recoveries ranged between 4.4-10.8 and 33-82%, respectively. The suggested method was linear (r(2) ≥0.997) over a dynamic range of 0.02-0.80 µg mL(-1) in plasma. The intra- and inter-days relative standard deviation (RSD%) and relative error (RE%) values of the method were below 20%, which shows good precision and accuracy. Finally, this method was applied to the analysis of real plasma samples obtained from the patients treated with these drugs.

  6. Unconventional High-Performance Laser Protection System Based on Dichroic Dye-Doped Cholesteric Liquid Crystals

    Science.gov (United States)

    Zhang, Wanshu; Zhang, Lanying; Liang, Xiao; Le Zhou; Xiao, Jiumei; Yu, Li; Li, Fasheng; Cao, Hui; Li, Kexuan; Yang, Zhou; Yang, Huai

    2017-01-01

    High-performance and cost-effective laser protection system is of crucial importance for the rapid advance of lasers in military and civilian fields leading to severe damages of human eyes and sensitive optical devices. However, it is crucially hindered by the angle-dependent protective effect and the complex preparation process. Here we demonstrate that angle-independence, good processibility, wavelength tunability, high optical density and good visibility can be effectuated simultaneously, by embedding dichroic anthraquinone dyes in a cholesteric liquid crystal matrix. More significantly, unconventional two-dimensional parabolic protection behavior is reported for the first time that in stark contrast to the existing protection systems, the overall parabolic protection behavior enables protective effect to increase with incident angles, hence providing omnibearing high-performance protection. The protective effect is controllable by dye concentration, LC cell thickness and CLC reflection efficiency, and the system can be made flexible enabling applications in flexible and even wearable protection devices. This research creates a promising avenue for the high-performance and cost-effective laser protection, and may foster the development of optical applications such as solar concentrators, car explosion-proof membrane, smart windows and polarizers. PMID:28230153

  7. MYCOBACTERIUM COMPLEX IDENTIFICATION BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

    Directory of Open Access Journals (Sweden)

    S.A HAWAII

    2001-12-01

    Full Text Available Introduction: There are different ways for identification of Mycobacteria. One of the most sensitive method is HPLC of phenacyl esters of mycolic acids of Mycobacteria for rapid identification of them after their primary cultures. This study uses HPLC for rapid identification and dissociation of Mycobacterium tuberculosis complex. Methods: In this study we use HPLC patterns of mycolic acids for identification three important species of mycobacteria (M. tuberculosis, M. bovis, M. bovis BCG from other mycobacterial species. All the strains were obtained from Tuberculosis and Pulmonary Diseases Research Center. HPLC conditions was as follows: HPLC: Model 1200 Cecil, Column: URP C-18 25X4.6 mm, Detector: U.V variable wave length at 254 nm, Elution: Gradient of methanol/chloroform. Flow rate: 2.5 ml/min. Results: HPLC leads to obtaining chromatograms which on its X-axis retention times (of different peaks which exist in the sample and on its Y-axis U.V absorbance (of these peaks were drown. These chromatograms in M. bovis and M. tuberculosis samples are similar with each other but differs from BCG ones. Discussion: On the basis of different retention times and numbers of the peaks which present in each chromatogram, we can differentiate between M. bovis, M. tuberculosis and BCG from other Mycobacteria. Also, with this method we can identify BCG from M. bovis and M. tuberculosis (because BCG has 9 and M. bovis and M. tuberculosis has 7 characteristic peaks in their chromatograms.

  8. [Determination of benproperine phosphate tablets by high performance liquid chromatography].

    Science.gov (United States)

    Yang, X J; Yang, X Y

    2000-11-01

    A reversed-phase HPLC method for the determination of benproperine phosphate tablets is reported. The chromatographic conditions were Hypersil-C18 column(5 microns, 4.6 mm i.d. x 150 mm) at 35 degrees C, mobile phase of methanol-water-glacial acetic acid-triethylamine(60:35:5:0.1, V/V) with a flow rate of 0.9 mL/min and UV detection at 270 nm. Cortisone acetate was selected as the internal standard. The linear relationship of calibration curve was good in the range of 9.96 mg/L-49.8 mg/L(r = 0.9998). The average recovery and RSD were 99.91%(n = 5) and 0.43% respectively. The total time of analysis for a run was within 7 min. The method is simple, sensitive, rapid and accurate.

  9. HIGH PERFORMANCE LIQUID CHROMATOGRAPHY DETECTORS – A REVIEW

    Directory of Open Access Journals (Sweden)

    Kaushal Ramni

    2011-05-01

    Full Text Available HPLC is the most versatile and widely used elution chromatography. The technique is used to resolve and determine species in a variety of organic, inorganic, biological, ionic and polymeric materials. Detector is the heart of an instrument and efficiency of system is dependent upon detecting techniques. Many types of HPLC detectors exist, each of which has some valuable performance feature such as refractive index detector, ultraviolet detector, fluorescent detector, electrochemical detector, electric conductivity detector, liquid light scattering detector, evaporative light scattering detector. Due to strong requirement for improvements in sensitivity, selectivity and other performance characteristics of the detector recent developments in conventional techniques and some other new technologies have been adopted such as laser light scattering detector, charged aerosol detector, nano quantity aerosol detector, chiral detector and pulsed amperometric detector. These detectors provide accurate concentration analysis, excellent sensitivity, wide dynamic range, consistent response and broad applicability of the drug components. Working of these detectors involve different principles such as optical techniques, aerosol based techniques, refractive methods, light scattering principle, amperometric and fluorescence. The present review enlightens both conventional and advanced techniques and compares their capabilities of analyzing drug components and need for new techniques for better and wide range of applicability.

  10. Ultrasound-assisted dispersive liquid-liquid microextraction combined with high-performance liquid chromatography-fluorescence detection for sensitive determination of biogenic amines in rice wine samples.

    Science.gov (United States)

    Huang, Ke-Jing; Wei, Cai-Yun; Liu, Wei-Li; Xie, Wan-Zhen; Zhang, Jun-Feng; Wang, Wei

    2009-09-18

    Ultrasound-assisted dispersive liquid-liquid microextraction coupled with high-performance liquid chromatography-fluorescence detection was used for the extraction and determination of three biogenic amines including octopamine, tyramine and phenethylamine in rice wine samples. Fluorescence probe 2,6-dimethyl-4-quinolinecarboxylic acid N-hydroxysuccinimide ester was applied for derivatization of biogenic amines. Acetonitrile and 1-octanol were used as disperser solvent and extraction solvent, respectively. Extraction conditions including the type of extraction solvent, the volume of extraction solvent, ultrasonication time and centrifuging time were optimized. After extraction and centrifuging, analyte was injected rapidly into high-performance liquid chromatography and then detected with fluorescence. The calibration graph of the proposed method was linear in the range of 5-500 microg mL(-1) (octopamine and tyramine) and 0.025-2.5 microg mL(-1) (phenethylamine). The relative standard deviations were 2.4-3.2% (n=6) and the limits of detection were in the range of 0.02-5 ng mL(-1). The method was applied to analyze the rice wine samples and spiked recoveries in the range of 95.42-104.56% were obtained. The results showed that ultrasound-assisted dispersive liquid-liquid microextraction was a very simple, rapid, sensitive and efficient analytical method for the determination of trace amount of biogenic amines.

  11. Rapid and Green Analytical Method for the Determination of Quinoline Alkaloids from Cinchona succirubra Based on Microwave-Integrated Extraction and Leaching (MIEL Prior to High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Farid Chemat

    2011-11-01

    Full Text Available Quinas contains several compounds, such as quinoline alkaloids, principally quinine, quinidine, cinchonine and cichonidine. Identified from barks of Cinchona, quinine is still commonly used to treat human malaria. Microwave-Integrated Extraction and Leaching (MIEL is proposed for the extraction of quinoline alkaloids from bark of Cinchona succirubra. The process is performed in four steps, which ensures complete, rapid and accurate extraction of the samples. Optimal conditions for extraction were obtained using a response surface methodology reached from a central composite design. The MIEL extraction has been compared with a conventional technique soxhlet extraction. The extracts of quinoline alkaloids from C. succirubra obtained by these two different methods were compared by HPLC. The extracts obtained by MIEL in 32 min were quantitatively (yield and qualitatively (quinine, quinidine, cinchonine, cinchonidine similar to those obtained by conventional Soxhlet extraction in 3 hours. MIEL is a green technology that serves as a good alternative for the extraction of Cinchona alkaloids.

  12. Rapid and green analytical method for the determination of quinoline alkaloids from Cinchona succirubra based on Microwave-Integrated Extraction and Leaching (MIEL) prior to high performance liquid chromatography.

    Science.gov (United States)

    Fabiano-Tixier, Anne-Sylvie; Elomri, Abdelhakim; Blanckaert, Axelle; Seguin, Elisabeth; Petitcolas, Emmanuel; Chemat, Farid

    2011-01-01

    Quinas contains several compounds, such as quinoline alkaloids, principally quinine, quinidine, cinchonine and cichonidine. Identified from barks of Cinchona, quinine is still commonly used to treat human malaria. Microwave-Integrated Extraction and Leaching (MIEL) is proposed for the extraction of quinoline alkaloids from bark of Cinchona succirubra. The process is performed in four steps, which ensures complete, rapid and accurate extraction of the samples. Optimal conditions for extraction were obtained using a response surface methodology reached from a central composite design. The MIEL extraction has been compared with a conventional technique soxhlet extraction. The extracts of quinoline alkaloids from C. succirubra obtained by these two different methods were compared by HPLC. The extracts obtained by MIEL in 32 min were quantitatively (yield) and qualitatively (quinine, quinidine, cinchonine, cinchonidine) similar to those obtained by conventional Soxhlet extraction in 3 hours. MIEL is a green technology that serves as a good alternative for the extraction of Cinchona alkaloids.

  13. Optimized determination of polybrominated diphenyl ethers by ultrasound-assisted liquid-liquid extraction and high-performance liquid chromatography.

    Science.gov (United States)

    He, Kuang; Lv, YuanCai; Chen, YuanCai

    2014-10-01

    A method based on ultrasound-assisted liquid-liquid extraction and high-performance liquid chromatography has been optimized for the determination of six polybrominated diphenyl ether congeners. The optimal condition relevant to the extraction was first investigated, more than 98.7 ± 0.7% recovery was achieved with dichloromethane as extractant, 5 min extraction time, and three cycles of ultrasound-assisted liquid-liquid extraction. Then multiple function was employed to optimize polybrominated diphenyl ether detection conditions with overall resolution and chromatography signal area as the responses. The condition chosen in this experiment was methanol/water 93:7 v/v, flow rate 0.80 mL/min, column temperature 30.0°C. The optimized technique revealed good linearity (R(2) > 0.9962 over a concentration range of 1-100 μg/L) and repeatability (relative standard deviation liquid-liquid extraction coupled with high-performance liquid chromatography was effective to identify and quantify the complex polybrominated diphenyl ethers in effluent samples.

  14. Rapid method for monitoring N-nitrosodimethylamine in drinking water at the ng/L level without pre-concentration using high-performance liquid chromatography-chemiluminescence detection.

    Science.gov (United States)

    Kodamatani, Hitoshi; Yamasaki, Hitomi; Sakaguchi, Takeru; Itoh, Shinya; Iwaya, Yoshimi; Saga, Makoto; Saito, Keiitsu; Kanzaki, Ryo; Tomiyasu, Takashi

    2016-08-19

    As a contaminant in drinking water, N-nitrosodimethylamine (NDMA) is of great concern because of its carcinogenicity; it has been limited to levels of ng/L by regulatory bodies worldwide. Consequently, a rapid and sensitive method for monitoring NDMA in drinking water is urgently required. In this study, we report an improvement of our previously proposed HPLC-based system for NDMA determination. The approach consists of the HPLC separation of NDMA, followed by NDMA photolysis to form peroxynitrite and detection with a luminol chemiluminescence reaction. The detection limit for the improved HPLC method was 0.2ng/L, which is 10 times more sensitive than our previously reported system. For tap water measurements, only the addition of an ascorbic acid solution to eliminate residual chlorine and passage through an Oasis MAX solid-phase extraction cartridge are needed. The proposed NDMA determination method requires a sample volume of less than 2mL and a complete analysis time of less than 15min per sample. The method was utilized for the long-term monitoring of NDMA in tap water. The NDMA level measured in the municipal water survey was 4.9ng/L, and a seasonal change of the NDMA concentration in tap water was confirmed. The proposed method should constitute a useful NDMA monitoring method for protecting drinking water quality. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Ultrasound-assisted magnetic dispersive solid-phase microextraction: A novel approach for the rapid and efficient microextraction of naproxen and ibuprofen employing experimental design with high-performance liquid chromatography.

    Science.gov (United States)

    Ghorbani, Mahdi; Chamsaz, Mahmoud; Rounaghi, Gholam Hossein

    2016-03-01

    A simple, rapid, and sensitive method for the determination of naproxen and ibuprofen in complex biological and water matrices (cow milk, human urine, river, and well water samples) has been developed using ultrasound-assisted magnetic dispersive solid-phase microextraction. Magnetic ethylendiamine-functionalized graphene oxide nanocomposite was synthesized and used as a novel adsorbent for the microextraction process and showed great adsorptive ability toward these analytes. Different parameters affecting the microextraction were optimized with the aid of the experimental design approach. A Plackett-Burman screening design was used to study the main variables affecting the microextraction process, and the Box-Behnken optimization design was used to optimize the previously selected variables for extraction of naproxen and ibuprofen. The optimized technique provides good repeatability (relative standard deviations of the intraday precision 3.1 and 3.3, interday precision of 5.6 and 6.1%), linearity (0.1-500 and 0.3-650 ng/mL), low limits of detection (0.03 and 0.1 ng/mL), and a high enrichment factor (168 and 146) for naproxen and ibuprofen, respectively. The proposed method can be successfully applied in routine analysis for determination of naproxen and ibuprofen in cow milk, human urine, and real water samples.

  16. Hydrocarbon group type determination in jet fuels by high performance liquid chromatography

    Science.gov (United States)

    Antoine, A. C.

    1977-01-01

    Results are given for the analysis of some jet and diesel fuel samples which were prepared from oil shale and coal syncrudes. Thirty-two samples of varying chemical composition and physical properties were obtained. Hydrocarbon types in these samples were determined by fluorescent indicator adsorption (FIA) analysis, and the results from three laboratories are presented and compared. Recently, rapid high performance liquid chromatography (HPLC) methods have been proposed for hydrocarbon group type analysis, with some suggestion for their use as a replacement of the FIA technique. Two of these methods were used to analyze some of the samples, and these results are also presented and compared. Two samples of petroleum-based Jet A fuel are similarly analyzed.

  17. DETERMINATION OF FLUCONAZOLE IN HUMAN PLASMA BY REVERSE PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHY.

    Science.gov (United States)

    Safaei, Zahra; Alipour, Eskandar; Shafaati, Alireza; Zarghp, Afshin

    2015-01-01

    A rapid, simple and reproducible high performance liquid chromatographic method was developed and validated for determination of fluconazole in human plasma. The separation was performed on MZ C8 column (125 x 4 mm, 5 µm) using acetonitrile - potassium dihydrogen phosphate buffer (15 : 85, v/v), pH 3.0, as the mobile phase at a flow rate of 1.5 mL/min. The wavelength was set at 261 nm. The assay enables the measurement of fluconazole for therapeutic drug monitoring with a minimum quantification limit of 20 ng/mL. The method involves simple, protein precipitation procedure and analytical recovery was complete. The calibration curve was linear over the concentration range 0.1-4 µg/mL. The coefficients of variation for inter-day and intra-day assay were found to be less than 10%.

  18. Application of High Performance Liquid Chromatography to Separation of Novel Chiral Tetrahedral Heterometal Clusters

    Institute of Scientific and Technical Information of China (English)

    Xin Yi ZHU; Wei Qiang ZHANG; Yu Hua ZHANG; Li Ren CHEN; Yong Min LI

    2003-01-01

    A series of novel chiral tetrahedral heterometal clusters have firstly been separated oncellulose tris-(3,5-dimethylphenylcarbamate) stationary phase by high performance liquid chrom-atography, using hexane as the mobile phase with various alcohols as modifiers.

  19. Development and validation of high performance liquid chromatographic method for analysis of clozapine.

    Science.gov (United States)

    Kaur, Harpreet; Bassi, Pallavi; Monif, Tausif; Khuroo, Arshad; Kaur, Gurpreet

    2013-05-01

    In this study a rapid, simple and sensitive assay to quantify clozapine in human plasma by using reverse phase high performance liquid chromatographic method has been developed. Clozapine was extracted from human plasma using a mixture of chloroform: n-hexane 50:50 employing liquid-liquid extraction method. The calibration curve was found to be linear in the concentration range of 25-800 ng/ml. The inter day and intra day assay accuracy and precision fulfilled the criteria specified by USFDA, Guidance for industry: bioanalytical method validation. Clozapine was found to be stable in human plasma after 6 h incubation at room temperature, 50 days storage at -27°C and freeze thaw cycles, as well as after reconstitution with mobile phase after 24 h of storage in refrigerator. The validated method offers the advantage of using minimum injection volume (25μl) and plasma sample volume (300μl). The extraction method is simple and single step with no back extraction step, thus, making this method applicable to determination of pharmacokinetic profiles and parameters.

  20. Determination of diflubenzuron and chlorbenzuron in fruits by combining acetonitrile-based extraction with dispersive liquid-liquid microextraction followed by high-performance liquid chromatography.

    Science.gov (United States)

    Ruan, Chunqiang; Zhao, Xiang; Liu, Chenglan

    2015-09-01

    In this study, a simple and low-organic-solvent-consuming method combining an acetonitrile-partitioning extraction procedure followed by "quick, easy, cheap, effective, rugged and safe" cleanup with ionic-liquid-based dispersive liquid-liquid microextraction and high-performance liquid chromatography with diode array detection was developed for the determination of diflubenzuron and chlorbenzuron in grapes and pears. Ionic-liquid-based dispersive liquid-liquid microextraction was performed using the ionic liquid 1-hexyl-3-methylimidazolium hexafluorophosphate as the extractive solvent and acetonitrile extract as the dispersive solvent. The main factors influencing the efficiency of the dispersive liquid-liquid microextraction were evaluated, including the extractive solvent type and volume and the dispersive solvent volume. The validation parameters indicated the suitability of the method for routine analyses of benzoylurea insecticides in a large number of samples. The relative recoveries at three spiked levels ranged between 98.6 and 109.3% with relative standard deviations of less than 5.2%. The limit of detection was 0.005 mg/kg for the two insecticides. The proposed method was successfully used for the rapid determination of diflubenzuron and chlorbenzuron residues in real fruit samples.

  1. Determination of sulfonamides in butter samples by ionic liquid magnetic bar liquid-phase microextraction high-performance liquid chromatography.

    Science.gov (United States)

    Wu, Lijie; Song, Ying; Hu, Mingzhu; Xu, Xu; Zhang, Hanqi; Yu, Aimin; Ma, Qiang; Wang, Ziming

    2015-01-01

    A novel, simple, and environmentally friendly pretreatment method, ionic liquid magnetic bar liquid-phase microextraction, was developed for the determination of sulfonamides in butter samples by high-performance liquid chromatography. The ionic liquid magnetic bar was prepared by inserting a stainless steel wire into the hollow of a hollow fiber and immobilizing ionic liquid in the micropores of the hollow fiber. In the extraction process, the ionic liquid magnetic bars were used to stir the mixture of sample and extraction solvent and enrich the sulfonamides in the mixture. After extraction, the analyte-adsorbed ionic liquid magnetic bars were readily isolated with a magnet from the extraction system. It is notable that the present method was environmentally friendly since water and only several microliters of ionic liquid were used in the whole extraction process. Several parameters affecting the extraction efficiency were investigated and optimized, including the type of ionic liquid, sample-to-extraction solvent ratio, the number of ionic liquid magnetic bars, extraction temperature, extraction time, salt concentration, stirring speed, pH of the extraction solvent, and desorption conditions. The recoveries were in the range of 73.25-103.85 % and the relative standard deviations were lower than 6.84 %. The experiment results indicated that the present method was effective for the extraction of sulfonamides in high-fat content samples.

  2. Determination of dodecylbenzenesulphonates and ethoxylated alkylphenols in liquid pesticide formulations by high-performance liquid chromatography.

    Science.gov (United States)

    Schreuder, R H; Martijn, A

    1988-01-01

    A method is presented for determining mixtures of calcium dodecylbenzene-sulphonate (CaDBS) and ethoxylated alkylphenols in pesticidal emulsifiable concentrates. The pesticide formulations are adsorbed on a pre-concentration column. The active ingredients, the solvents and the emulsifiers are eluted successively by using solvents with increasing polarities. The various homologues of CaDBS and the ethoxylated alkylphenols are separated by reversed-phase high performance liquid chromatography using methanol-water containing tetramethylammonium bromide. The oligomers of the ethoxylated phenols are separated on an aminopropyl-modified column using a solvent gradient (propan-2-ol-water and hexane-tetrahydrofuran). Quantitative results are given for synthetic samples. The degree of ethoxylation of the ethoxylated alkylphenols has been determined and the CaDBS components characterized.

  3. High performance liquid-level sensor based on mPOFBG for aircraft applications

    DEFF Research Database (Denmark)

    Marques, C. A. F.; Pospori, A.; Saez-Rodriguez, D.

    2015-01-01

    A high performance liquid-level sensor based on microstructured polymer optical fiber Bragg grating (mPOFBG) array sensors is reported in detail. The sensor sensitivity is found to be 98pm/cm of liquid, enhanced by more than a factor of 9 compared to a reported silica fiber-based sensor....

  4. 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.

  5. Rapid detection of malto-oligosaccharide-forming bacterial amylases by high performance anion-exchange chromatography

    DEFF Research Database (Denmark)

    Duedahl-Olesen, Lene; Larsen, K. L.; Zimmermann, W.

    2000-01-01

    High performance anion-exchange chromatography with pulsed amperometric detection was applied for the rapid analysis of malto-oligosaccharides formed by extracellular enzyme preparations from 49 starch-degrading bacterial strains isolated from soil and compost samples. Malto-oligosaccharide-formi...

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

    Science.gov (United States)

    Wang, Ye; Tian, Minglei; Bi, Wentao; Row, Kyung Ho

    2009-01-01

    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. PMID:19582220

  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. A Method for the Quantitation of Trace Levels of Dimethyl Sulfoxide in Urine by High Performance Liquid Chromatography

    Science.gov (United States)

    1989-05-01

    HIGH PERFORMANCE LIQUID CHROMATOGRAPHY by...for the sample cleanup and concentration, followed by separation by reversed phase high performance liquid chromatography . EXPERIMENTAL Materials...DIMETHYL SULFOXIDE IN URINE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY 4. AUTHORS (Last name, first name, middle initial. If military, show rank, e.g.

  9. Determination of Dimethyl Sulfoxide (DMSO), Ethanol (ETOH), Formamide (F) and Glycerol/Formal (GF) by High Performance Liquid Chromatography (HPLC)

    Science.gov (United States)

    1989-01-30

    HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC...Classification) (U) Determination of Dimethyl Sulfoxide (DMSO), Ethanol, (ETOH), Formamide (F), and Glycerol/ Formal (GF) by High Performance Liquid Chromatography (HPLC...and 5). High performance liquid chromatography (HPLC) was the analytical method of choice for analyzing DMSO, ethanol, formamide and

  10. Determination of 5-Bromo-2’-Deoxyuridine (BrdU) in Well Water by High Performance Liquid Chromatography (HPLC)

    Science.gov (United States)

    1992-09-01

    HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC...Securrty Classification) Determination of 5-Bromo-2’-Deoxyuridine (BrdU) in Well Water by High Performance Liquid Chromatography (hPLC) 12. PERSONAL...PLOT OF BrdU STABILITY VERSUS TIME ....................... 10 ii DETERMINATION OF 5-BROMO-2’-DEOXY-URIDINE (BrdU) IN WELL WATER BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

  11. 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 C18 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.

  12. [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.

  13. DETERMINATION OF CARBENDAZIM IN WATER BY HIGH-PERFORMANCE IMMUNOAFFINITY CHROMATOGRAPHY ON-LINE WITH HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH DIODE-ARRAY OR MASS SPECTROMETRIC DETECTION

    Science.gov (United States)

    An automated method for the determination of carbendazim in water that combines high-performance immunoaffinity chromatography (HPIAC), high-performance liquid chromatography (HPLC) in the reversed-phase mode, and detection by either UV-Vis diode array detector (DAD) spectroscopy...

  14. 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.

  15. Ultrapreconcentration and determination of organophosphorus pesticides in water by solid-phase extraction combined with dispersive liquid-liquid microextraction and high-performance liquid chromatography.

    Science.gov (United States)

    Chen, Junhua; Zhou, Guangming; Deng, Yongli; Cheng, Hongmei; Shen, Jie; Gao, Yi; Peng, Guilong

    2016-01-01

    Solid-phase extraction coupled with dispersive liquid-liquid microextraction was developed as an ultra-preconcentration method for the determination of four organophosphorus pesticides (isocarbophos, parathion-methyl, triazophos and fenitrothion) in water samples. The analytes considered in this study were rapidly extracted and concentrated from large volumes of aqueous solutions (100 mL) by solid-phase extraction coupled with dispersive liquid-liquid microextraction and then analyzed using high performance liquid chromatography. Experimental variables including type and volume of elution solvent, volume and flow rate of sample solution, salt concentration, type and volume of extraction solvent and sample solution pH were investigated for the solid-phase extraction coupled with dispersive liquid-liquid microextraction with these analytes, and the best results were obtained using methanol as eluent and ethylene chloride as extraction solvent. Under the optimal conditions, an exhaustive extraction for four analytes (recoveries >86.9%) and high enrichment factors were attained. The limits of detection were between 0.021 and 0.15 μg/L. The relative standard deviations for 0.5 μg/L of the pesticides in water were in the range of 1.9-6.8% (n = 5). The proposed strategy offered the advantages of simple operation, high enrichment factor and sensitivity and was successfully applied to the determination of four organophosphorus pesticides in water samples.

  16. 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 ...

  17. Monitoring stevioside in soju by high-performance liquid chromatography and liquid chromatography/mass spectrometry.

    Science.gov (United States)

    Ni, Fan; Ammann, Jeffrey; Mabud, Abdul

    2007-01-01

    A method using high-performance liquid chromatography (HPLC) with UV absorption detection was developed to monitor stevioside in soju, a distilled spirits product that is commercially available. The method uses a single-step dilution for sample preparation. It completely eliminates the time-consuming process of solid-phase extraction. A method using HPLC/mass spectrometry was optimized to confirm the identities of stevioside and other related impurities, including rebaudioside A, rebaudioside C, and dulcoside. The method was validated. The validation parameters included range (10.1-1007.3 ppm), precision, linearity, accuracy, robustness, system suitability, and intermediate precision. Stevioside standard solutions at 6 concentration levels were prepared for the validation work, including the tests for precision, linearity, and accuracy. The solutions were prepared in triplicate for each concentration. The relative standard deviation for the precision test was 0.999. The average recovery ranged from 95.7 to 101.1% for the soju samples spiked with stevioside standard. The detection limit for stevioside was estimated at 75 ppb. The method was used to screen several soju samples; no detectable stevioside was found in the samples.

  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. Ionic liquids for improving the extraction of NSAIDs in water samples using dispersive liquid-liquid microextraction by high performance liquid chromatography-diode array-fluorescence detection.

    Science.gov (United States)

    Toledo-Neira, Carla; Álvarez-Lueje, Alejandro

    2015-03-01

    A rapid, sensitive and efficient analytical method based on the use of ionic liquids for determination of non-steroidal anti-inflammatory drugs (NSAIDs) in water samples was developed. High-performance liquid chromatography equipped with a diode array and fluorescence detector was used for quantification of ketoprofen, ibuprofen and diclofenac in tap and river water samples. This new method relies on the use of two ionic liquids with multiple functionalities: one functions as an extraction solvent (1-butyl-3-methylimidazolium hexafluorophosphate ([BMIM][PF6]), and the other changes the polarity in the aqueous medium (1-butyl-3-methylimidazolium tetrafluoroborate, ([BMIM][BF4]). Factors such as the type and volume of the ILs and dispersive solvent, sample volume, and centrifugation time were investigated and optimized. The optimized method exhibited good precision, with relative standard deviation values between 2% and 3%, for the three NSAIDs. Limits of detection achieved for all of the analytes were between 17 and 95 ng mL(-1), and the recoveries ranged from 89% to 103%. Furthermore, the enrichment factors ranged from 49 to 57. The proposed method was successfully applied to the analysis of NSAIDs in tap and river water samples.

  20. 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.

  1. High-performance liquid chromatographic assay for metamizol metabolites in rat plasma: application to pharmacokinetic studies.

    Science.gov (United States)

    Domínguez-Ramírez, Adriana Miriam; Calzadilla, Patricia Carrillo; Cortés-Arroyo, Alma Rosa; Hurtado Y de la Peña, Marcela; López, José Raúl Medina; Gómez-Hernández, Martín; López-Muñoz, Francisco Javier

    2012-12-01

    In order to evaluate the pharmacokinetics of metamizol in the presence of morphine in arthritic rats, after subcutaneous administration of the drugs, an easy, rapid, sensitive and selective analytical method was proposed and validated. The four main metamizol metabolites (4-methylaminoantipyrine, 4-aminoantipyrine, 4-acetylaminoantipyrine and 4-formylaminoantipyrine) were extracted from plasma samples (50-100μl) by a single solid-phase extraction method prior to reverse-phase high performance liquid chromatography with diode-array detection. Standard calibration graphs for all metabolites were linear within a range of 1-100μg/ml (r(2)≥0.99). The intra-day coefficients of variation (CV) were in the range of 1.3-8.4% and the inter-day CV ranged from 1.5 to 8.4%. The intra-day assay accuracy was in the range of 0.6-9.6% and the inter-day assay accuracy ranged from 0.9 to 7.5% of relative error. The lower limit of quantification was 1μg/ml for all metabolites using a plasma sample of 100μl. Plasma samples were stable at least for 4 weeks at -20°C. This method was found to be suitable for studying metamizol metabolites pharmacokinetics in arthritic rats, after simultaneous administration of metamizol and morphine, in single dose. Copyright © 2012 Elsevier B.V. All rights reserved.

  2. [Determination of five coumarins in toys by high performance liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Yang, Rongjing; Wei, Biwen; Gao, Huan; Yu, Wenjia

    2012-02-01

    A rapid analytical method for the determination of five coumarins (coumarin, 7-methoxycoumarin, dihydrocoumarin, 7-methyl coumarin and 7-ethoxy-4-methyl coumarin) in toys by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) has been developed. After ultrasonic extraction in tetrahydrofuran, the samples were analyzed by HPLC-MS/MS in multi-reaction monitoring (MRM) mode. Acetonitrile and 0.1% acetic acid were used as the mobile phases with gradient elution. The linear ranges of calibration curves were 10 - 1 000 microg/L, and the limits of quantification (LOQ) (S/N > 10) were 2.0 microg/L for all the analytes, except that the LOQ for dihydrocoumarin was 5.0 microg/L. The recoveries of the five coumarins spiked in three types of samples were in the ranges of 93.2% - 105.8%, 97.3% - 103.2% and 96.8% - 102.9%, with the relative standard deviations in the ranges of 4.35% - 8.27%, 3.65% - 6.73% and 4.03% - 6.45%, respectively. The method was applied in the determination of 12 toy samples. The five analytes were found in 9 samples, and in some cases, the presence of quite high concentrations of these coumarins in the toys should be a matter of concern.

  3. Determination of xanthohumol in beer based on cloud point extraction coupled with high performance liquid chromatography.

    Science.gov (United States)

    Chen, Ligang; Zhao, Qi; Jin, Haiyan; Zhang, Xiaopan; Xu, Yang; Yu, Aimin; Zhang, Hanqi; Ding, Lan

    2010-04-15

    A method based on coupling of cloud point extraction (CPE) with high performance liquid chromatography separation and ultraviolet detection was developed for determination of xanthohumol in beer. The nonionic surfactant Triton X-114 was chosen as the extraction medium. The parameters affecting the CPE were evaluated and optimized. The highest extraction yield of xanthohumol was obtained with 2.5% of Triton X-114 (v/v) at pH 5.0, 15% of sodium chloride (w/v), 70 degrees C of equilibrium temperature and 10 min of equilibrium time. Under these conditions, the limit of detection of xanthohumol is 0.003 mg L(-1). The intra- and inter-day precisions expressed as relative standard deviations are 4.6% and 6.3%, respectively. The proposed method was successfully applied for determination of xanthohumol in various beer samples. The contents of xanthohumol in these samples are in the range of 0.052-0.628 mg L(-1), and the recoveries ranging from 90.7% to 101.9% were obtained. The developed method was demonstrated to be efficient, green, rapid and inexpensive for extraction and determination of xanthohumol in beer.

  4. Sequence-dpenedent DNA separation by anion-exchange high-performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Yamakawa, Hisashi; Higashino, Ken-ich; Ohara, Osamu [Kazusa DNA Research Inst., Chiba (Japan)

    1996-09-05

    High-performance liquid chromatography (HPLC) system with a new nonporous anion-exchange resin, DNA-NPR, made it possible to rapidly separate DNA fragments up to 20 kbp with high resolution. In order to further characterize this chromatographic DNA separation system, we prepared a mixtures of double-stranded DNAs of constant length carrying a fully degenerated 50-bp region and analyzed their chromatographic behavior on the DNA-NPR column. The results indicated that the separation of DNA fragments on the anion-exchange HPLC was governed not only by size, but also by nucleotide sequence: even DNA fragments with the same size and the same base content could be separated on this column. Taking advantage of this characteristic feature of the anion-exchange HPLC, we could readily fractionate human cDNAs with practically acceptable recovery and high resolution. Furthermore, the combination of HPLC and gel electrophoresis realized separation of a mixture of DNA fragments in a two-dimensional pattern. 22 refs., 5 figs., 1 tab.

  5. 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: 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.

  6. Molecular monitoring of the intestinal flora by denaturing high performance liquid chromatography.

    Science.gov (United States)

    Goldenberg, Oliver; Herrmann, Stefanie; Marjoram, Gina; Noyer-Weidner, Mario; Hong, George; Bereswill, Stefan; Göbel, Ulf B

    2007-01-01

    Gut flora analysis is hampered by the complexity of the intestinal microbiota and by inherent limitations of culture-based approaches. Therefore, culture-independent molecular methods based upon 16S rRNA gene analysis were applied successfully for the analysis of complex microbial communities. However, generally accepted and validated profiling methods such as denaturing and temperature gradient gel electrophoresis (DGGE/TGGE) are still laborious and time consuming. Thus, we adapted the separation of amplified bacterial 16S rRNA gene fragments by denaturing high performance liquid chromatography (DHPLC) using the WAVE Microbial Analysis System as a rapid and convenient means to display complex intestinal bacterial communities and to monitor changes in the gut flora. The separation of 16S rRNA gene fragments amplified from reference strains representing main gut bacterial populations and from human stool samples revealed that DHPLC analysis effectively detects bacterial groups predominant in the human gut flora. The investigation of faecal samples from hospitalized patients before, during and after antibiotic therapy showed that PCR-based DHPLC can be used to monitor gut flora changes. Results from DHPLC analysis were comparable with DGGE profiles generated from the same samples, demonstrating that the adapted DHPLC protocol is well suited for the analysis of complex microbial communities.

  7. Determination of Duloxetine and Its Major Metabolites in Rabbit Plasma by High-Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    T. K. Laha

    2015-03-01

    Full Text Available A rapid and sensitive high performance liquid chromatographic method is described for simultaneous determination of duloxetine and its major metabolites, such as 4- hydroxy duloxetine (M7, Glucuronide conjugate of 5-hydroxy-6-methoxy duloxetine (M6 and Glucuronide conjugate of dihydrodiol duloxetine (M12 in rabbit plasma. HPLC analysis was carried out on a µ-Bondapak C18 column (250 mm × 4.6 mm, 5µm particle size using methanol: phosphate buffer (pH 7.9, 50 mM (7:3 v/v as the mobile phase at a flow rate of 1.5 ml/min. Detection was carried out at 224 nm with an UV detector. The above metabolites present in the rabbit plasma were characterized by retro-synthesis followed by NMR and MS study for structure confirmation and finally injected separately into the HPLC system. All the three retention time matches with the metabolites present in the plasma sample.

  8. REVERSE PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC METHOD FOR THE ANALYSIS OF GLIPIZIDE IN PHARMACEUTICAL DOSAGE FORMS

    Directory of Open Access Journals (Sweden)

    Sheikh Rahila

    2010-12-01

    Full Text Available A rapid and sensitive reverse phase high performance liquid chromatographic methods depicted for the qualitative and quantitative assay of glipizide in pharmaceutical dosage forms. Glipizide was chromatographed on reverse phase C18 column with mobile phase consisting of 0.05 M Potassium Dihydrogen Orthophosphate: Methanol [15: 85 %v/v, pH 7.0 ± 0.05, adjusted with 1% Triethylamine]. The mobile phase was pumped at a flow rate 1 mL/min. Quantification was achieved by monitoring the ultraviolet absorbance at 225 ηm. The average retention time for Glipizide was found to be 3.21 ± 0.07. With this method, linearity was observed in the range of 10 – 2000 ηg/ml. The LOD and LOQ were found to be 5 ηg/ml and 15 ηg/ml respectively. The method was applicable for the analysis of drug in tablet formulation. The results of analysis were validated statistically.

  9. Enantioselective high-performance liquid chromatographic determination of nicardipine in human plasma.

    Science.gov (United States)

    Uno, T; Ohkubo, T; Sugawara, K

    1997-09-26

    A sensitive method for the enantioselective high-performance liquid chromatography (HPLC) determination of nicardipine in human plasma is described. (+)-Nicardipine, (-)-nicardipine and (+)-barnidipine as an internal standard are detected by an ultraviolet detector at 254 nm. Racemic nicardipine in human plasma was extracted by a rapid and simple procedure based on C18 bonded-phase extraction. The extraction samples were purified and concentrated on a pre-column using a C1 stationary phase and the enantiomers of nicardipine are quantitatively separated by HPLC on a Sumichiral OA-4500 column, containing a chemically modified Pirkle-type stationary phase. Determination of (+)- and (-)-nicardipine was possible in a concentration range of 5-100 ng ml(-1) and the limit of detection in plasma was 2.5 ng ml(-1). The recoveries of (+)- and (-)-nicardipine added to plasma were 91.4-98.4% and 93.3-96.7%, respectively, with coefficients of variation of less than 9.0 and 9.4% respectively. The method was applied to low level monitoring of (+)- and (-)-nicardipine in plasma from healthy volunteers.

  10. [Simultaneous determination of 10 synthetic colorants in cosmetics by high performance liquid chromatography].

    Science.gov (United States)

    Liu, Haishan; Qian, Xiaoyan; Lü, Chunhu; Zhu, Xiaoyu; Chen, Xiaomei; Mo, Weimin

    2013-11-01

    A high performance liquid chromatographic method was developed for the simultaneous determination of 10 synthetic colorants in cosmetics. The cosmetics were extracted by the ultrasonic technique with tetrahydrofuran (THF), dimethyl sulfoxide (DMSO) and methanol sequentially. Then the extracts were centrifuged for purification and separated on an Eclipse XDB-C18 column (150 mm x 4.6 mm, 5 microm) with gradient elution by acetonitrile and 0.02 mol/L ammonium acetate (pH 4.60, adjusted with acetic acid). A diode array detector was used to determine the colorants with the wavelengths ranging from 417 nm to 640 nm. The linear relationships of the 10 colorants between the peak areas and the mass concentrations were obtained in the range of 0.5-20.0 mg/L (r > 0.999). The limits of quantitation ranged from 10 to 20 mg/kg. The average recoveries at three concentration levels ranged from 92.9% to 108.8% with the relative standard deviations in the range of 0.5% to 6.1% (n = 6). The method is simple, rapid and sensitive. It is suitable for the simultaneous determination of the 10 colorants in the oil cosmetics, cream cosmetics and powder cosmetics.

  11. Determination of Residual Acrylamide in Medical Polyacrylamide Hydrogel by High Performance Liquid Chromatography tandem Mass Spectroscopy

    Institute of Scientific and Technical Information of China (English)

    WEI-WEI LI; HUI LI; ZHI-FEI LIU; QUN QIAO

    2009-01-01

    Objective To determine residual acrylamide in medical polyacrylamide hydrogel by high performance liquid chromatography tandem mass spectroscopy (HPLC-MS). Methods After 13C3 labeled acrylamide was added, the sample was extracted with water and then cleaned up with ExtrelutTM 20. The polyaerylamide hydrogel sample and 20 clinical cases were analyzed by HPLC-MS/MS and isotope dilution quantifying technique in selected reaction monitoring (SRM) mode. Results Acrylamide was separated from polyacrylamide hydrogel. The concentration of acrylamide in polyacrylamide hydrogel ranged from 3.9×109 to 3.1×108g/L in the 20 clinical cases. The peak area was favorable linear and the range was up to 3 000 μg/L. The recovery rate was 103.1% with a relative standard deviation (RSD) of 6.20%, when the mark level was 50 μg/L. Conclusion HPLC-MS is a rapid, accurate, and sensitive method for the determination of residual acrylamide in medical polyacrylamide hydrogel.

  12. Simultaneous Determination of Potassium Sorbate and Sodium Benzoate in Doogh Using High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    S. Sohrabvandi

    2015-09-01

    Full Text Available In this research, high performance liquid chromatography (HPLC as a simple and rapid technique was used for the determination of potassium sorbate and sodium benzoate in different brands of Doogh (typical Iranian drink based on fermented milk. The chromatographic separation was achieved with a C18 column (250 mm×4 I.D, 5 µm and one buffered mobile phase, acetate buffer (0.2 mol/L, pH4.4: acetonitrile (63:37 at a flow rate of 1 mL/min. The effluent was monitored at 235nm, respectively. The main affecting factors, such as Carrez solution amount, pH and centrifuge speed were optimized. Limits of detection (LODs were 1.823 for sorbic acid and 1.985 for benzoic acid. Relative standard deviations (RSDs for quintuplicate analyses were 4.4 for sorbic acid and 4.087 for benzoic acid. The method also showed good linearity in a range from 4 to 1000 mg/L with correlation coefficients (R2 of 0.990 for sorbic acid and 0.995 for benzoic acid. Recoveries for the two analytes were 80 for sorbic acid and 65 for benzoic acid. Comparing to the other techniques, the proposed method was significantly less time-consuming and had more acceptable accuracy, precision and interruption elimination in samples. The developed method was successfully used for the routine monitoring of benzoate and sorbate in Doogh.

  13. Spectrophotometric and high performance liquid chromatographic methods for sensitive determination of bisphenol A.

    Science.gov (United States)

    Zhuang, Yafeng; Zhou, Meng; Gu, Jia; Li, Xiangmei

    2014-03-25

    A new spectrophotometric method for the determination of trace amounts of bisphenol A based on a diazotization-coupling reaction was developed. In acidic solution, clenbuterol was first diazotized with sodium nitrite, then coupled with bisphenol A to from an azo-compound [I] in NH3-NH4Cl buffer, which shows a maximum absorption at 410 nm. The effects of the amount of sodium nitrite, diazo reaction time, the amount of clenbuterol, coupling reaction time and coupling reaction temperature have been examined. Under the optional conditions, the determination of the linear range of bisphenol A is 0.24-8.4 μg/mL, correlation coefficient is 0.9905 and detection limit of this method is 0.15 μg/mL. The spectrophotometric method is simple, rapid, high sensitivity with better accuracy. High performance liquid chromatography (HPLC) technique combined with this new spectrophotometric method has been also developed for the measurement of bisphenol A. The analysis was achieved on a C18 column using water and methanol as a mobile phase and the detection was done spectrophotometrically at 410 nm. These reported methods were applied to the determination of bisphenol A in hot water in contact with commercially available table-water bottle samples.

  14. A simple high-performance liquid chromatographic practical approach for determination of flurbiprofen

    Directory of Open Access Journals (Sweden)

    Muhammad Akhlaq

    2011-01-01

    Full Text Available A simple, rapid, sensitive, and specific high-performance liquid chromatography (HPLC assay for flurbiprofen has been developed and validated practically. The chromatography was conducted using Gemini C18 column (5 μm; 4.6 mm × 250 mm, Phenomenex, California, USA. The mobile phase containing disodium hydrogen phosphate solution (30 mM pH 7.0 and acetonitrile (50:50; and the isocratic flow rate of 1.0 ml/min were used in the current study. Detection was made at 247 nm. The calibration curve was linear (r ≥ 0.9996 over the concentration range of 5-50 mm/ml. Mean percentage (% recovery ± % relative standard deviation (RSD ranged from 97.07 ± 0.008 to 103.66 ± 0.013. Within-day and between-day precision were also in acceptable range of 98.83 ± 0.004 to 104.56 ± 0.009. In order to confirm the practical applicability of the method developed, flurbiprofen controlled release matrix tablets were subjected to the dissolution studies and the release rate was analyzed. The reported HPLC for flurbiprofen provides several advantages of simplicity, high specificity, accuracy, and very short run-cycle time. It is suggested that the method should be used for the routine quality control analysis of flurbiprofen pure drug and its dosage forms.

  15. Quantitation of clobazam in human plasma using high-performance liquid chromatography.

    Science.gov (United States)

    Knapp, J; Bokník, P; Gumbinger, H G; Linck, B; Lüss, H; Müller, F U; Schmitz, W; Vahlensieck, U; Neumann, J

    1999-05-01

    A rapid and simple reversed-phase high-performance liquid chromatographic (HPLC) method for the determination of clobazam concentrations in human blood samples is developed and validated. Solid-phase column extraction is performed to clean up blood samples before running the analytical HPLC system. The chromatography is isocratic with a mobile phase consisting of acetonitrile (20%, v/v), methanol (23%, v/v), and 0.1 M potassium hydrogen phosphate buffer (pH 3.6; 57%, v/v) at a constant flow rate of 2 mL/min. Clobazam is detected at 226 nm. Chromatography is completed within less than 25 min. The recovery rate is greater than 95% and linear over a wide range of drug concentrations. The intra-assay coefficient of variation percentage varies between 4.3 and 12. This method is used for therapeutic drug monitoring in patients undergoing antiepileptic therapy with clobazam. Plasma levels of clobazam ranged from 21 to 663 ng/mL. Other antiepileptic compounds, such as clonazepam and phenobarbital, did not interfere with the detection of clobazam.

  16. Determination of hexamethylenetetramine in foods by high-performance liquid chromatography (HPLC).

    Science.gov (United States)

    Lim, HoSoo; Kim, JungIm; Ko, KyungYuk; Kim, Meehye

    2014-01-01

    A simple and rapid method was developed and validated for the determination of hexamethylenetetramine (HMT) in foods. Samples were homogenised and extracted with methanol, followed by centrifugation. The resulting solution was filtered and injected into the high-performance liquid chromatograph (HPLC). HMT was separated using a Zorbax SCX-300 column coupled to a photodiode array detector. The calibration curve was linear in the range of 1.0-100 μg ml(-1), with good correlation coefficients (r(2) = 0.9992). The recoveries of HMT from foods spiked at levels of 10, 50 and 100 mg kg(-1) ranged from 91.6% to 103.8%, with relative standard deviations (RSDs) between 0.9% and 5.3%. The limit of detection and the limit of quantification of HMT were 0.3 and 1.0 mg kg(-1) based on three food matrixes (provolone cheese, glass noodle and tofu snack), respectively. Uncertainty associated with accuracy contributed mostly to the expanded uncertainty. No detectable levels of HMT were found in any of the samples retailed in Korea. The method was successful in determining HMT in foods.

  17. Detection of synthetic corticosteroids in bovine urine by chemiluminescence high-performance liquid chromatography.

    Science.gov (United States)

    Vázquez, B I; Feás, X; Lolo, M; Fente, C A; Franco, C M; Cepeda, A

    2005-01-01

    The development of a black market of chemical cocktails for illegal growth promotion in food-producing animals includes substances that are potentially dangerous for human health, such as synthetic corticosteroids. The potential presence of these residues in food makes it necessary to develop rapid and sensitive analytical methodologies to detect such substances, preferably in live animals before they arrive at the market. A chemiluminescence (CL) detection method for the determination of four synthetic corticosteroids (prednisolone, betamethasone, dexamethasone and flumethasone) in bovine urine has been developed. The proposed system, which does not need any derivatization procedure, offers an easy method well suited for routine research. Urine samples were homogenized with methanol:water (50:50, v/v) and centrifuged. The upper layer was collected and Strata X cartridges were used for cleaning up. The purified residues were evaporated to dryness and then redissolved in the mobile phase. Analysis of the extracts was performed using high-performance liquid chromatography with chemiluminescence detection, employing luminol as the CL reagent. The recovery curves, obtained at four spiking levels (different for each corticosteroid), showed that recoveries of at least 70% could be obtained for urine. The chemiluminescence detection procedure afforded satisfactory results with respect to sensitivity and the LOD and LOQ, taken as the first point of the regression curve, ranged from 4 ppb to 65 ppb. The maximum mean RSD was below 13% and below 15% for intra- and inter-day assay, respectively, in all cases.

  18. 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.

  19. DETERMINATION OF ENZYME-ACTIVITY BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

    NARCIS (Netherlands)

    WELLING, GW; SCHEFFER, AJ; WELLINGWESTER, S

    1994-01-01

    The application of high-performance liquid chromatography (HPLC) in the study of enzymatic reactions is reviewed. The rationale for using HPLC is given and whether the components of the reaction mixture should be derivatized prior to or after HPLC. An alphabetical list of enzymes assayed by HPLC is

  20. 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...

  1. Going Beyond, Going Further: An Inexpensive Experiment for the Introduction of High Performance Liquid Chromatography.

    Science.gov (United States)

    Bidlingmeyer, Brian A.; Warren, F. Vincent, Jr.

    1984-01-01

    Background information, materials needed, laboratory procedures, and typical results are provided for five high performance liquid chromatography experiments (three isocratic and two step gradient separations). Suggestions for further experimentation are also provided, including quantitative determinations and separation of charged solutes. (JN)

  2. 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…

  3. 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…

  4. 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...

  5. 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.

  6. 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…

  7. 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 extrapo

  8. Determination of rafoxanide and closantel in ovine plasma by high performance liquid chromatography.

    Science.gov (United States)

    Benchaoui, H A; McKellar, Q A

    1993-01-01

    A high performance liquid chromatographic method has been developed for the determination of rafoxanide and closantel in ovine plasma. Acetonitrile and chloroform were used for the extraction. The mean recoveries were 78.69% and 80.59% for rafoxanide and closantel, respectively. This method was applied to the characterization of rafoxanide plasma kinetics following oral administration of therapeutic doses to sheep.

  9. Comparison of high performance liquid chromatography and enzymatic analysis of soluble carbohydrates in loblolly pine

    Science.gov (United States)

    Patricia L. Faulkner; Michele M. Schoeneberger; Kim H. Ludovici

    1993-01-01

    Foliar tissue was collected from a field study designed to test impacts of atmospheric pollutants on loblolIy pine (Pinus taeda L.) seedlings. Standard enzymatic (ENZ) and high performance liquid chromatography (HPLC) methods were used to analyze the tissue for soluble sugars. A comparison of the methods revealed no significant diffennces in accuracy...

  10. New methods and materials for solid phase extraction and high performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Dumont, Philip John [Iowa State Univ., Ames, IA (United States)

    1996-04-23

    This paper describes methods for solid phase extraction and high performance liquid chromatography (HPLC). The following are described: Effects of Resin Sulfonation on the Retention of Polar Organic Compounds in Solid Phase Extraction; Ion-Chromatographic Separation of Alkali Metals In Non-Aqueous Solvents; Cation-Exchange Chromatography in Non-Aqueous Solvents; and Silicalite As a Stationary Phase For HPLC.

  11. 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…

  12. Quantification of six bioactive compounds in Zhenqi Fuzheng preparation by high-performance liquid chromatography couple

    Institute of Scientific and Technical Information of China (English)

    Yi-Kai Shi; Fang Cui; Fang-Di Hu; Ying-Yan Bi; Yu-Feng Ma; Shi-Lan Feng

    2011-01-01

    A simple and accurate high-performance liquid chromatography(HPLC)coupled with diode array detector(DAD)and evaporative light scattering detector(ELSD)was established for the determination of six bioactive compounds in Zhenqi Fuzheng preparation(ZFP).The

  13. 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…

  14. High Performance Liquid Chromatographic Analysis of Phytoplankton Pigments Using a C16-Amide Column

    Science.gov (United States)

    A reverse-phase high performance liquid chromatographic (RP-HPLC) method was developed to analyze in a single run, most polar and non-polar chlorophylls and carotenoids from marine phytoplankton. The method is based on a RP-C16-Amide column and a ternary gradient system consistin...

  15. 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…

  16. High-performance liquid chromatography determination of dipotassium clorazepate and its major metabolite nordiazepam in plasma.

    Science.gov (United States)

    Colin, P; Sirois, G; Lelorier, J

    1983-04-08

    A rapid and sensitive high-performance liquid chromatographic method is described for the quantitative analysis of dipotassium clorazepate (CZP) and its major metabolite nordiazepam (ND) in fresh human and dog plasma. The method consists of two separate selective ND extractions from a plasma sample without and with conversion of all the CZP to ND. For quantitation, diazepam (DZP) is used as the internal standard. The chromatographic phase utilized in a reversed-phase Hibar EC-RT analytical column prepacked with LiChrosolv RP-18 with a solvent system consisting of acetonitrile-0.05 M sodium acetate buffer, pH 5.0 (45:55). The UV absorbance is monitored at 225 nm using a variable-wavelength detector. The mean assay coefficient of variation over a concentration range of 20-400 ng per ml of plasma is less than 3% for the within-day precision. Recoveries of ND, DZP and CZP (as ND) are essentially quantitative at all levels investigated. The calibration curves of ND are rectilinear (r2 = 0.99) from the lower limit of sensitivity (2 ng/ml) to at least 2000 ng/ml in plasma. Applicability of the method to CZP and ND disposition studies in the anaesthetized mongrel dog is illustrated. When the two separate selective nordiazepam extractions from plasma cannot be performed immediately after blood sampling, an extrapolation kinetic method is suggested for the estimation of CZP concentration. In all previous in vivo studies, CZP has been determined only with gas-liquid chromatographic methods.

  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. 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.

  19. Rapid screening of transferrin-binders in the flowers of Bauhinia blakeana Dunn by on-line high-performance liquid chromatography-diode-array detector-electrospray ionization-ion-trap-time-of-flight-mass spectrometry-transferrin-fluorescence detection system.

    Science.gov (United States)

    Liu, Meixian; Dong, Jing; Lin, Zongtao; Niu, Yanyan; Zhang, Xiaotian; Jiang, Haixiu; Guo, Ning; Li, Wei; Wang, Hong; Chen, Shizhong

    2016-06-10

    Transferrin (Transferrin, TRF, TF) has drawn increasing attention in cancer therapy due to its potential applications in drug delivery. TF receptor, highly expressed in tumor cells, recognizes and transports Fe(3+)-TF into cells to release iron into cytoplasm. Thus, discovering TF-binding compounds has become an active research area and is of great importance for target therapy. In this study, an on-line analysis method was established for screening TF-binding compounds from the flowers of Bauhinia blakeana Dunn using a high-performance liquid chromatography-diode-array detector-multi-stage mass spectrometry-transferrin-fluorescence detector (HPLC-DAD-MS(n)-TF-FLD) method. As a result, 33 of 80 identified or tentatively characterized compounds in the sample were TF-binding active. Twenty-five flavonol glycosides and eight phenolic acids were identified as TF-binders. Twelve of these active compounds together with six standard compounds were used to study the dose-response effects and structure-activity relationships of flavonoids and phenolic acids. The method was validated by vitexin with a good linearity in the range of concentrations used in the study. The limit of detection for vitexin was 0.1596 nmol. Our study indicated that the established method is simple, rapid and sensitive for screening TF-binding active compounds in the extract of Bauhinia blakeana Dunn, and therefore is important for discovering potential anti-cancer ingredients from complex samples for TF related drug delivery.

  20. Using a single tablet daily to treat latent tuberculosis infection in Brazil: bioequivalence of two different isoniazid formulations (300 mg and 100 mg demonstrated by a sensitive and rapid high-performance liquid chromatography-tandem mass spectrometry method in a randomised, crossover study

    Directory of Open Access Journals (Sweden)

    André Daher

    2015-06-01

    Full Text Available The recommended treatment for latent tuberculosis (TB infection in adults is a daily dose of isoniazid (INH 300 mg for six months. In Brazil, INH was formulated as 100 mg tablets. The treatment duration and the high pill burden compromised patient adherence to the treatment. The Brazilian National Programme for Tuberculosis requested a new 300 mg INH formulation. The aim of our study was to compare the bioavailability of the new INH 300 mg formulation and three 100 mg tablets of the reference formulation. We conducted a randomised, single dose, open label, two-phase crossover bioequivalence study in 28 healthy human volunteers. The 90% confidence interval for the INH maximum concentration of drug observed in plasma and area under the plasma concentration vs. time curve from time zero to the last measurable concentration “time t” was 89.61-115.92 and 94.82-119.44, respectively. The main limitation of our study was that neither adherence nor the safety profile of multiple doses was evaluated. To determine the level of INH in human plasma, we developed and validated a sensitive, simple and rapid high-performance liquid chromatography-tandem mass spectrometry method. Our results showed that the new formulation was bioequivalent to the 100 mg reference product. This finding supports the use of a single 300 mg tablet daily strategy to treat latent TB. This new formulation may increase patients’ adherence to the treatment and quality of life.

  1. Salting-out assisted liquid-liquid extraction coupled to dispersive liquid-liquid microextraction for the determination of chlorophenols in wine by high-performance liquid chromatography.

    Science.gov (United States)

    Fan, Yingying; Hu, Shibin; Liu, Shuhui

    2014-12-01

    A novel procedure of sample preparation combined with high-performance liquid chromatography with diode array detection is introduced for the analysis of highly chlorinated phenols (trichlorophenols, tetrachlorophenols, and pentachlorophenol) in wine. The main features of the proposed method are (i) low-toxicity diethyl carbonate as extraction solvent to selectively extract the analytes without matrix effect, (ii) the combination of salting-out assisted liquid-liquid extraction and dispersive liquid-liquid microextraction to achieve an enrichment factor of 334-361, and (iii) the extract is analyzed by high-performance liquid chromatography to avoid derivatization. Under the optimum conditions, correlation coefficients (r) were >0.997 for calibration curves in the range 1-80 ng/mL, detection limits and quantification limits ranged from 0.19 to 0.67 and 0.63 to 2.23 ng/mL, respectively, and relative standard deviation was wines, with recovery rates in the range 82-104%. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Vortex-assisted low density solvent based demulsified dispersive liquid-liquid microextraction and high-performance liquid chromatography for the determination of organophosphorus pesticides in water samples.

    Science.gov (United States)

    Seebunrueng, Ketsarin; Santaladchaiyakit, Yanawath; Srijaranai, Supalax

    2014-05-01

    A simple, rapid, effective and eco-friendly preconcentration method, vortex-assisted low density solvent based solvent demulsified dispersive liquid-liquid microextraction (VLDS-SD-DLLME), followed by high performance liquid chromatography-diode array detector (HPLC-DAD) analysis, has been developed for the first time for the determination of four organophosphorus pesticides (OPPs) (e.g., azinphos-methyl, parathion-methyl, fenitrothion and diazinon) in environmental water samples. In this preconcentration procedure, an emulsion was obtained after the mixture of extraction solvent (1-dodecanol) and dispersive solvent (acetonitrile, ACN) was injected rapidly into 10 mL of the sample solution. The vortex agitator aided the dispersion of the extraction solvent into the sample solution. After the formation of an emulsion, the demulsifier (ACN) was added, resulting in the rapid separation of the mixture into two phases without centrifugation. Under optimal conditions, the proposed method provided high extraction efficiency (90-99%), good linearity range (0.5-500 ng mL(-1)), low limits of detection (0.25-1 ng mL(-1)) and good repeatability and recoveries were obtained.

  3. 低温离心净化-高效液相色谱法快速测定腐竹中的碱性橙II和碱性嫩黄O%Rapid determination of chrysoidine II and auramine O in dried bean curd sticks by cryogenic centrifugal purification-high performance liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    杨爽; 俞子萱; 王永芳; 葛宝坤

    2015-01-01

    ABSTRACT:Objective Toestablish a method to detect chrysoidine II and auramine O in dried bean curd sticks by cryogenic centrifugal purification–high performance liquid chromatography. Methods The samples werediluted and extracted by 70% acetonitrile ammonia water, purified by centrifugation at 12000 r/min and 4℃, separated by high performance liquid chromatography (HPLC), and detected by photodiode array detector (PDA). The method took the retention time (Rt) for qualitative and external standard method quantitative analysis.Results Within the limits of concentration 0.00125~0.05 mg/mL, the standard curve had a good linear relationship, and the correlation coefficient was more than 0.9999; in the level of 0.05, 2.0, and 10.0 mg/kg, the add mark recovery rate was from 95.0% to 103.0%(n=10), the relative standard deviation was less than 5.0%; limit of detection(LOD) was 0.02 mg/kg, limit of quantitation (LOQ) was 0.05 mg/kg.Conclusion Comparing the actual sample testing results with reference method, this method is simple, rapid, sensitive, with low cost, and is suitable for popularization and application.%目的:建立腐竹中碱性橙 II 和碱性嫩黄 O 的低温离心净化-高效液相色谱测定方法。方法样品经70%的乙腈-氨水(0.5%)提取,4℃低温、12000 r/min离心净化,高效液相色谱(high performance liquid chromatography, HPLC)分离,最后采用二极管阵列检测器(PDA)检测,以保留时间(Rt)定性,外标法定量。结果在0.00125~0.05 mg/mL的浓度范围内,标准曲线有良好的线性关系,相关系数大于0.9999;样品在0.05、2.0、10.0 mg/kg三个加标水平内的回收率范围为95.0%~103.0%(n=10),相对标准偏差小于5.0%。方法的检出限(LOD)为0.02 mg/kg,定量限(LOQ)为0.05 mg/kg。结论对实际样品测试结果与参考文献方法比较,该方法具有简便、快速、灵敏、费用低,便于推广应用。

  4. Determination of furanochromones and pyranocoumarins in drugs and Ammi visnaga fruits by combined solid-phase extraction-high-performance liquid chromatography and thin-layer chromatography-high-performance liquid chromatography.

    Science.gov (United States)

    Zgórka, G; Dragan, T; Głowniak, K; Basiura, E

    1998-02-27

    A new, simple and rapid solid-phase extraction method for the determination of furanochromones and pyranocoumarins in Ammi visnaga L. fruits and pharmaceuticals by reversed-phase high-performance liquid chromatography (RP-HPLC) was developed. The isolation of compounds examined was carried out on octadecyl BakerBond SPE columns using various concentrations of methanol, acetonitrile and tetrahydrofuran in water. High and reproducible recoveries were obtained. To compare the results of quantitative analysis a preparative TLC procedure was also elaborated and carried out.

  5. [Determination of congo red in beef by high performance liquid chromatography-tandem quadrupole time of flight mass spectrometry and ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry].

    Science.gov (United States)

    Lin, Hui; Xu, Chunxiang; Yan, Chunrong; Zhang, Zheng; Wang, Suilou

    2013-09-01

    A method was developed for the determination of congo red in beef. The analyte was identified by high performance liquid chromatography-tandem quadrupole time of flight mass spectrometry (LC-QTOF MS) and quantitatively determined by ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry. After purified by liquid-liquid extraction, the congo red in the beef sample was separated on an Agilent ZORBAX Eclipse Plus C18 Rapid Resolution HD UPLC column (50 mm x 2.1 mm, 1.8 microm) HPLC , using 95% (volume percentage) methanol as the mobile phase at 0.2 mL/min. The detection was performed on an AB 4000 + triple quadrupole mass spectrometer utilizing electrospray ionization (ESI) interface operated in negative ion mode and multiple-reaction monitoring (MRM) mode. The results showed that the linear range of congo red mass concentration was 0.03 - 1 mg/L with the correlation coefficient of 0.999 8. The method had a good precision with the RSDs lower than 5% and the recoveries ranging from 88% to 91%. The limit of detection (LOD) of congo red was 0.01 mg/L. With good reproducibility, the method is simple, fast and effective for the determination of the illegally added congo red in beef and other meat products.

  6. High-performance liquid chromatography with photodiode array detection and chemometrics method for the analysis of multiple components in the traditional Chinese medicine Shuanghuanglian oral liquid.

    Science.gov (United States)

    Li, Bao Qiong; Chen, Jing; Li, Jiao Jiao; Wang, Xue; Zhai, Hong Lin; Zhang, Xiao Yun

    2015-12-01

    Shuanghuanlian oral liquid, a traditional Chinese medicine preparation, is a mixture of three herbs (Flos Lonicerae, Radix Scutellariae and Fructus Forsythiae). In this study, the quantitative analysis of three main active compounds, chlorogenic acid, forsythin and baicalin in samples from different manufacturers was performed rapidly by high-performance liquid chromatography coupled with photodiode array detection followed by Contour Projection coupled to stepwise regression treatment of the obtained three-dimensional spectra in which the partial overlap between adjacent target components existed. The method was validated for linearity (R>0.9940), precision (RSDliquid samples.

  7. 高效液相色谱法快速测定银杏叶提取物中的萜类内酯%Rapid Analysis of Terpene Lactones in Extract of Ginkgo biloba L.by High Performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    汪海峰; 鞠兴荣

    2000-01-01

    A new rapid analytical method was developed for the quantification of terpene lactones (bilobalide(BB) and ginkgolide A,B,C,J) in extract of Ginkgo biloba L. (EGb) using a liquid-liquid solvent extraction procedure followed by high performance liquid chromatography. EGb was dissolved in 30% ethanol and extracted with ether. After evaporation, the residue was then determinated by HPLC on a C18 column with methanol-water-orthophosphoric acid (25∶75∶0.1,V/V) as eluent and refractive index (RI) detection. Results showed that the excellent sample clean-up procedure is more simple and specific, and saves more time (less than 20 min) than any other methods that have been reported, and also leads to high recoveries (>99.0%) and low RSDs (<2.0%).The reproducible method is regarded to be very useful for evaluating the quality of extract of Ginkgo biloba L..%建立了液-液萃取后高效液相色谱法(HPLC)测定银杏叶提取物(EGb)中萜类内酯含量的快速方法.EGb样品溶于体积分数为30%的乙醇溶液后用乙醚萃取,有机相浓缩后的残留物以HPLC分析,组分的分离采用C18柱,以甲醇-水-磷酸(体积比为25:75:0.1)为流动相,示差检测器检测.结果表明,样品净化程序较已报道的方法更简便,选择性强、快速(少于20 min).该方法回收率大于99.0%、相对标准偏差小于2.0%、重现性好,可有效地用于银杏叶提取物的产品质量评价.

  8. Separation of Nitration By-Products in Commercial-Grade Trinitro-Toluene by High Performance Liquid Chromatography.

    Science.gov (United States)

    1982-06-01

    MRL-TN-464 SEPARATION OF NITRATION BY-PRODUCTS IN COMMERCIAL-GRADE TITR-TOLUENE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY Peter J. Sanders ABSTRACT...UNCLASSIFIED TITLE SEPARATION OF NITRATION BY-PRODUCTS IN COMMERCIAL-GRADE TRXNITRO-TOLUENE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY AUTHOR(S...PRODUCTS IN COMMERCIAL-GRADE TRINITRO-TOLUENE BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY 16 INTRODUCTION Fbr some time, a need has existed for the

  9. Separation of polyprenyl phosphate oligomerhomologues by reversed-phase ion-pair high-performance liquid chromatography.

    Science.gov (United States)

    Kozlov, Vyacheslav V; Danilov, Leonid L

    2012-01-01

    A simple and rapid method of reversed-phase ion-pair high-performance liquid chromatography (HPLC) with UV-detection on a common C(18) column in an isocratic mode with the addition of tetrabutylammonium phosphate for the separation of polyprenyl phosphate oligomerhomologues has been developed. The method was successfully applied to assay the composition of polyprenyl phosphates obtained from polyprenols isolated from mulberry (Morus alba) leaves and fir (Abies sibirica) needles (the active components of veterinary drugs Gamapren and Phosprenyl correspondingly).

  10. Dispersive liquid-liquid microextraction followed by high-performance liquid chromatography for determination of benzoate and sorbate in yogurt drinks and method optimization by central composite design.

    Science.gov (United States)

    Kamankesh, Marzieh; Mohammadi, Abdorreza; Tehrani, Zohreh Modarres; Ferdowsi, Roohallah; Hosseini, Hedayat

    2013-05-15

    A new method based on dispersive liquid-liquid microextraction (DLLME) followed by high-performance liquid chromatography (HPLC) for determination of benzoate and sorbate salts in yogurt drinks was developed. The effective parameters in DLLME process, including volume of extraction and disperser solvents, pH and salt effect, were optimized using response surface methodology (RSM) based on central composite design. The yogurt drink samples were extracted using NaOH and Carrez solutions (potassium hexaferrocyanide and zinc acetate) were used for sedimentation of proteins. For DLLME, a mixture of extraction solvent (1-octanol) and disperser solvent (ethanol) was rapidly injected into the sample solution by syringe and cloudy solution is formed. Subsequently, the upper 1-octanol layer was analyzed by HPLC. The detection limits for benzoate and sorbate were 0.06 ng mL(-1) and 0.15 ng mL(-1), respectively. The relative standard deviations (RSD) for seven analyses were 4.96% for benzoate and 4.58% for sorbate. The proposed method demonstrated good linearity and high enrichment factor. A clean separation and good chromatogram is readily achieved without the presence of matrix interference. A comparison of this method with previous methods demonstrated that the proposed method is an accurate, rapid and reliable sample-pretreatment method that gives very good enrichment factors and detection limits for extracting and determining sorbate and benzoate in yogurt drink samples.

  11. Ionic Liquids as Mobile Phase Additives for Separation of Nucleotides in High-Performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    ZHANG,Wen-Zhu(张文珠); HE,Li-Jun(何丽君); LIU,Xia(刘霞); JIANG,Sheng-Xiang(蒋生祥)

    2004-01-01

    Ionic liquids are a type of salts that are liquid at low temperature (< 100 ℃). Because of their some special properties, they have been widely used as new "green solvents" for many chemical reactions and liquid-liquid extraction in the past several years. In this paper, a new method for the separation of nucleotides is developed and the essential feature of the method is that 1-alkyl-3-methylimidazolium salts are used as mobile phase additives, resulting in a baseline separation of nucleotides without need of gradient elution and need of organic solvent addition as currently used in RP-HPLC. This study shows the potential application of ionic liquids as mobile phase additives in reversed-phase liquid chromatography.

  12. Determination of chlorophenols in honey samples using in-situ ionic liquid-dispersive liquid-liquid microextraction as a pretreatment method followed by high-performance liquid chromatography.

    Science.gov (United States)

    Fan, Chen; Li, Nai; Cao, Xueli

    2015-05-01

    In-situ ionic liquid-dispersive liquid-liquid microextraction (IL-DLLME) method was developed as a pretreatment method for the detection of six chlorophenols (CPs) in honey samples. The hydrophobic ionic liquid [C4MIM][NTf2], formed in-situ by the hydrophilic ionic liquid [C4MIM][BF4] and the ion exchange reagent LiNTf2 was used as the microextractant solvent of CPs from honey sample. Then the enriched analytes were back-extracted into 40 μL of 0.14 M NaOH solution and finally subjected to analysis by high-performance liquid chromatography. The method showed low limit of detection of CPs, 0.8-3.2 μg/L and high enrichment factor, 34-65 with the recoveries range from 91.60% to 114.33%. The method is simple, rapid, environmentally friendly and with high extraction efficiency.

  13. Analyses of acute kidney injury biomarkers by ultra-high performance liquid chromatography with mass spectrometry.

    Science.gov (United States)

    Al Za'abi, Mohammed; Ali, Badreldin H; ALOthman, Zeid A; Ali, Imran

    2016-01-01

    The newly developed acute kidney injury biomarkers are very important for the early and timely detection of kidney diseases. This review contains details of the analyses of several acute kidney injury biomarkers using ultra-high performance liquid chromatography-mass spectrometry in urine and plasma samples. In this review we attempt to discuss some aspects of the types of the biomarkers, patents, sample preparation, and the analyses. Besides, efforts were also made to discuss the possible uses of superficially porous (core-shell) columns in traditional and inexpensive high-performance liquid chromatography instruments. Additionally, the challenges and the future prospects are also highlighted. The present review will be useful for the academicians, scientists, and clinicians for the early detection of acute kidney injury biomarkers.

  14. Homochiral metal-organic framework used as a stationary phase for high-performance liquid chromatography.

    Science.gov (United States)

    Kong, Jiao; Zhang, Mei; Duan, Ai-Hong; Zhang, Jun-Hui; Yang, Rui; Yuan, Li-Ming

    2015-02-01

    Metal-organic frameworks are promising porous materials. Chiral metal-organic frameworks have attracted considerable attention in controlling enantioselectivity. In this study, a homochiral metal-organic framework [Co(2) (D-cam)(2) (TMDPy)] (D-cam = D-camphorates, TMDPy = 4,4'-trimethylenedipyridine) with a non-interpenetrating primitive cubic net has been used as a chiral stationary phase in high-performance liquid chromatography. It has allowed the successful separation of six positional isomers and six chiral compounds. The good selectivity and baseline separation, or at least 60% valley separation, confirmed its excellent molecular recognition characteristics. The relative standard deviations for the retention time of run-to-run and column-to-column were less than 1.8 and 3.1%, respectively. These results demonstrate that [Co(2) (D-cam)(2) (TMDPy)] may represent a promising chiral stationary phase for use in high-performance liquid chromatography.

  15. 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......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 establishing the ability of HPLC-LIF protein profiling technique for discrimination, using hard clustering and Fuzzy clustering methods. The clustering algorithms have quite successfully classified the profiles as belonging to normal, cancer of cervix, and oral cancer conditions....

  16. Estimation of loxoprofen sodium dihydrate in tablets by reverse phase high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    R. Nanthakumar

    2016-09-01

    Full Text Available A simple and rapid reverse phase high performance liquid chromatographic (RP-HPLC method has been developed and validated for the estimation of loxoprofen sodium dihydrate (LSD in tablets. An isocratic HPLC analysis was performed on Phenomenex, Luna C18(2 (250 × 4.6 mm, 5μ column. The compound was separated with the mixture of 0.4% orthophosphoric acid and acetonitrile in the ratio of 45:55 (v/v as a mobile phase at a flow rate of 1.0 mL/min. UV detection was performed at 210 nm. Run time per sample was 8 min with the retention time of 4.9 min. The system suitability parameters, such as theoretical plate count, tailing and% RSD between six standard injections were within the limits. The method was validated according to ICH guidelines. The method was validated for specificity, precision, linearity, stability of sample solution, intermediate precision, robustness and accuracy. The stability of the sample solution was checked for 24 h at one hour intervals. The results show that the sample solution is stable for at least 24 h. Calibration plots were linear over the concentration range of 10–90 μg/mL as indicated by the correlation coefficient of 0.9992. The% RSD was 0.115 and 1.288 for system precision and method precision, respectively. The % RSD values for intermediate precision studies were less than 2%. The robustness of the method was evaluated by deliberately altering the chromatographic conditions and the results were adhered with the limits. The high recovery and low relative standard deviation confirm the suitability of the method for the estimation of loxoprofen sodium dihydrate in tablets.

  17. [Determination of phospholipids in bear bile by isocratic high performance liquid chromatography].

    Science.gov (United States)

    Lu, X Q; Wang, Z H; Hong, X K; Sun, J J; Zhang, S W

    1999-11-01

    A high performance liquid chromatographic method for the determination of phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylinositol (PI) in the bear bile was established. The air-dried bile of bear was ground into powder and extracted continuously with V(chloroform):V(methanol) = 1:1 to a final dilution 25-fold the volume of the sample. The extract was washed by addition to it of 0.5 its volume of 0.58% aqueous NaCl. The resulting mixture separated into two phases. The lower phase was the total pure lipid extract and was diluted to the desired volume by addition of V(chloroform):V(methanol) = 1:1 mixture and then injected into the HPLC system. The conditions of HPLC were P-E silica column (150 mm x 4.6 mm i.d., 10 microns); V(acetonitrile):V(methanol) = 76:24 mobile phase with a flow rate of 1.5 mL/min and UV detector at 205 nm. The linear regression equation was Y = 5,917X + 1,709(r = 0.9970) for PI, Y = 7,362.1X-6,608.2(r = 0.9949) for PE and Y = 2,457X-2,460.5(r = 0.9972) for PC. The linear range were 1.0-12.0 micrograms, 2.0-24.0 micrograms and 9.5-114.0 micrograms respectively, The relative standard deviation of peaks were 1.6%, 0.89% and 2.6% respectively. The average recovery of PC was 89.30% (RSD = 2.0%). The method enables a simple, rapid and reproducible quantification of PI, PE and PC.

  18. The Application of High Performance Liquid Chromatography in the Analysis of Trace Rare Earth

    Institute of Scientific and Technical Information of China (English)

    WANG; Xiu-feng; DING; You-qian

    2012-01-01

    <正>Rare earth elements are very important in the field of radioanalytical chemistry, for it must be separated and determined in the measurements of burn-up and fission yield. High performance liquid chromatography has become a main method in the separation of rare earth elements due to its obvious advantages, this is, high speed of analysis, high efficiency and easy automation. The ion exchange chromatography is the main means to separate rare earth elements, especially the cation exchange

  19. 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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Weatherby, R.P.; Allan, R.D.; Johnston, G.A.R. (Sydney Univ. (Australia))

    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.

  1. A simple high performance liquid chromatography method for determination of rebamipide in rat urine

    OpenAIRE

    Cooper, Dustin L.; Harirforoosh, Sam

    2014-01-01

    Rebamipide is a mucoprotective agent commonly used to prevent nonsteriodal anti-inflammatory drug-induced gastrointenstinal side effects [1]. Human plasma and urine analysis of rebamipide utilizing high performance liquid chromatography (HPLC) have been reported [2]. Recently, we reported on the plasma levels of rebamipide in presense or absence of celecoxib or diclofenac in rats [3] using a modified HPLC method of detection developed by Jeoung et al. [4]. To tailor the method towards use in ...

  2. Quality Evaluation of Potentilla fruticosa L. by High Performance Liquid Chromatography Fingerprinting Associated with Chemometric Methods

    OpenAIRE

    Wei Liu; Dongmei Wang; Jianjun Liu; Dengwu Li; Dongxue Yin

    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, Sichua...

  3. [Comparison of refractometric and ultraviolet detection methods in lactulose assay by high performance liquid chromatography].

    Science.gov (United States)

    Vaccher, C; Berthelot, P; Flouquet, N; Debaert, M

    1990-01-01

    Dosage of lactulose and determination of related sugars (lactose, fructose, galactose, épilactose) were undertaken by high performance liquid chromatography (HPLC). The present note described two modes of detection used simultaneously and compared: refractive index detector (RI) and multiwavelength ultraviolet detector (UV). HPLC is performed with an aminopropylsylil phase as stationary phase and an isocratic mobile phase containing acetonitrile and aqueous sodium phosphate. The results are outlined and discussed.

  4. DETERMINATION OF PROTEINS FROM BUFFALO MILK USING HIGH PERFORMANCE LIQUID CROMATOGRAPHY RP-HPLC

    OpenAIRE

    AURELIA PECE; COROIAN C.; BIANCA GHIRILĂ; G. MURESAN; VIOARA MIRESAN

    2013-01-01

    In the hereby paper, we have undertaken a study on buffalo milk proteins, employing high performance liquid chromatography (HPLC). This RP-HPLC technique is commonly employed in the separation and assessment of caseins K, and in the fresh, as well as processed milk. These methods are also successfully applied in the authenticity and origin assessment of certain cheese products and the qualitative analysis of milk in bubalines, ovines, caprines and bovines (Ferreira si Cacote, 2003; Veloso si...

  5. Surface Confined Ionic Liquid-A New Stationary Phase for the Separation of Ephedrines in High-performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    Shu Juan LIU; Feng ZHOU; Xiao Hua XIAO; Liang ZHAO; Xia LIU; Sheng Xiang JIANG

    2004-01-01

    In this article, a new and effective stationary phase based on ionic liquid modified silica is first reported and used for the separation of ephedrines in high-performance liquid chromatography (HPLC). The separation results indicate the high efficiency and reproducibility of the stationary phase. The electrostatic interaction, ion-exchange interaction between the solutes and the stationary phase are considered to attribute the effective separation. Moreover, the free silanols on the surface of the silica are effectively masked by the immobilized ionic liquid, a result of which is to decrease the non-specific absorption.

  6. Isolation and purification of heroin from heroin street samples by preparative high performance liquid chromatography.

    Science.gov (United States)

    Guo, Zhen; Zheng, Hui; Lu, Yanzhen; Wei, Yun

    2012-09-10

    The present study established a novel method using preparative high performance liquid chromatography to isolate and purify heroin·HCl from heroin street samples to be used as a reference standard. Different kinds of mobile phases and columns were used, ultimately the mobile phase consisting of hexane-isopropanol-methanol (65:28:7, v/v) and the SIL preparative column prepared in laboratory were selected as the final condition. Heroin was further purified by the drowning-out crystallization method using isopropanol-methanol (50:1, v/v) and hexane as drowning-out anti-solvents and salting-out agents, respectively. The purity was assessed by analytical high performance liquid chromatography and the confirmation of the chemical structure was performed by IR and NMR. About 110.7mg of heroin·HCl at a purity of over 99.52% was obtained from 180mg of heroin street samples which contained 156.15mg of heroin·HCl component by preparative high performance liquid chromatography. This method is suitable for preparing heroin standards in forensic science area.

  7. Fast and comprehensive analysis of secondary metabolites in cocoa products using ultra high-performance liquid chromatography directly after pressurized liquid extraction.

    Science.gov (United States)

    Damm, Irina; Enger, Eileen; Chrubasik-Hausmann, Sigrun; Schieber, Andreas; Zimmermann, Benno F

    2016-08-01

    Fast methods for the extraction and analysis of various secondary metabolites from cocoa products were developed and optimized regarding speed and separation efficiency. Extraction by pressurized liquid extraction is automated and the extracts are analyzed by rapid reversed-phase ultra high-performance liquid chromatography and normal-phase high-performance liquid chromatography methods. After extraction, no further sample treatment is required before chromatographic analysis. The analytes comprise monomeric and oligomeric flavanols, flavonols, methylxanthins, N-phenylpropenoyl amino acids, and phenolic acids. Polyphenols and N-phenylpropenoyl amino acids are separated in a single run of 33 min, procyanidins are analyzed by normal-phase high-performance liquid chromatography within 16 min, and methylxanthins require only 6 min total run time. A fourth method is suitable for phenolic acids, but only protocatechuic acid was found in relevant quantities. The optimized methods were validated and applied to 27 dark chocolates, one milk chocolate, two cocoa powders and two food supplements based on cocoa extract.

  8. Analysis of drugs of abuse in human plasma by dispersive liquid-liquid microextraction and high-performance liquid chromatography.

    Science.gov (United States)

    Fernández, P; Regenjo, M; Bermejo, A M; Fernández, A M; Lorenzo, R A; Carro, A M

    2015-04-01

    Opioids and cocaine are widely used at present, both for recreational purposes and as drugs of abuse. This raises the need to develop new analytical methods specifically designed for the simultaneous detection of several drugs of abuse in biological samples. In this work, dispersive liquid-liquid microextraction (DLLME) was assessed as a new sample treatment for the simultaneous extraction of morphine (MOR), 6-acetylmorphine (6AM), cocaine (COC), benzoylecgonine (BZE) and methadone (MET) from human plasma. Preliminary assays were done before developing an experimental design based on a Uniform Network Doehlert which allowed the optimum extraction conditions to be identified, namely: a volume of extractant solvent (chloroform) and dispersant solvent (acetonitrile) of 220 µl and 3.2 ml, respectively; 0.2 g of NaCl as a salting-out additive; pH 10.6 and ultrasound stirring for 3.5 min. The resulting extracts were analyzed by high-performance liquid chromatography with photodiode array detection (HPLC-PDA), using an XBridge® RP18 column (250 × 4.6 mm i.d., 5 µm particle size). Calibration graphs were linear over the concentration range 0.1-10 µg ml⁻¹, and detection limits ranged from 13.9 to 28.5 ng ml⁻¹. Precision calculated at three different concentration levels in plasma was included in the range 0.1-6.8% RSD. Recoveries of the five drugs were all higher than 84% on average. Finally the proposed method was successfully applied to 22 plasma samples from heroin, cocaine and/or methadone users, and the most frequently detected drug was benzoylecgonine, followed by methadone, cocaine and morphine.

  9. 分散固相萃取-高效液相色谱-串联质谱快速测定鸡肉中盐霉素残留%Rapid Determination of Salinomycin Residues in Chicken by Dispersive Solid Phase Extraction-High Performance Liquid Chromatography Tandem Mass Spectrometry

    Institute of Scientific and Technical Information of China (English)

    杨旺火; 吴少明; 周鹏; 刘飞

    2014-01-01

    A method for rapid determination of salinomycin in chicken was developed by ultra high performance liquid chromatography-electrospray ionization tandem mass spectrometry with dispersive solid-phase extraction. The compounds were extracted from chicken tissue with acetonitrile and then cleaned up by dispersive solid phase extraction, after solvent exchanged, the salinomycin was analyzed by LC-MS/MS on an Acquity BEH C18 column with methanol-0.3%FA-H2O solution as mobile phase by gradient elution under positive ion multiple reaction monitoring(MRM)mode. It showed, under optimized conditions the limit of detection for salinomycin was 0.03μg·kg-1, the recoveries between 5.5~31.4μg·kg-1spiked levels ranged from 78.2-97.8%, with RSDs less than 2.0%(n=6). The calibration curves was linear in the ranges of 0.630~25.2 μg·L-1, with a correlation coefficient more than 0.9989. Owe to its simple pretreatment, high sensitivity, this method could be used for rapid screening for detection of large quantities of samples.%建立了分散固相萃取-超高效色谱-电喷雾串联质谱快速测定鸡肉中盐霉素残留的方法。样品经乙腈提取后,采用QuEChERS方法净化,氮吹转换溶剂后,采用Acquity BEH C18柱进行分离,0.3%甲酸水溶液-甲醇体系为流动相,梯度洗脱,电喷雾质谱正离子多反应监测模式测定,外标法定量。结果表明,在优化的条件下对样品进行分析检测,盐霉素的检出限为0.03μg·kg-1,添加水平为5.5~31.4μg·kg-1时,加标回收率为78.2-97.8%,RSD0.9989。该方法前处理步骤简单,具有较高灵敏度,可用于大批量样品的快速筛查检测。

  10. Rapid Screening of Lipase Inhibitors from Fructus aurantii Using Lipase-immobilized Magnetic Beads Coupled with High-performance Liquid Chromatography-Mass Spectrometry%磁珠富集与液相色谱-质谱联用结合筛选枳壳中脂肪酶抑制剂

    Institute of Scientific and Technical Information of China (English)

    陈锥; 陶益; 王怡; 王毅

    2012-01-01

    提出了一种磁珠收集与液相色谱-质谱联用(LC-MS)结合用于快速筛选中药提取物中的脂肪酶抑制剂的方法.通过制备键合脂肪酶的磁珠,将其与枳壳总黄酮孵育后,筛选出枳壳中脂肪酶的配体.通过LC-MS分析发现,4个化合物均是潜在的脂肪酶抑制剂,鉴定其分别为柚皮苷、新橙皮苷、橙皮苷和枸橘苷.对4个化合物进行了体外脂肪酶抑制活性验证.研究结果表明,新橙皮苷、橙皮苷和枸橘苷具有显著的脂肪酶抑制活性,IC50分别为48.04,52.45和46.18 μg/mL,其中枸橘苷具有脂肪酶抑制活性为首次报道.结果表明,磁珠富集与LC-MS集成技术能够用于快速发现中药活性成分.%A new method for the rapidly screening of lipase inhibitors from extracts of traditional Chinese medicine using lipase-immobilized magnetic beads coupled with high-performance liquid chromatography-mass spectrometry(LC-MS) was proposed. After the lipase-immobilized magnetic beads were prepared, they were incubated with total flavonoid of Fructus aurantii to fish the ligands of lipase. Then, the eluent of magnetic beads' screening was analyzed by LC-MS. Four compounds, as naringin, hesperidin, neohesperidin and pon-cirin, were identified as the potential lipase inhibitors. Furthermore, the in vitro inhibitory activities against lipase of four compounds were tested. The results showed that hesperidin, neohesperidin and poncirin exhibit lipase inhibitory activities, while poncirin was firstly reported to be a lipase inhibitor. Our research showed that active compounds could be rapidly screened from extracts of traditional Chinese medicine via lipase-immobilized magnetic beads coupled with LC-MS.

  11. Preliminary Validation of High Performance Liquid Chromatography Method for Detection of Methyltestosterone Residue in Carp Muscle

    Institute of Scientific and Technical Information of China (English)

    JIANG Jie; LIN Hong; FU Xiaoting; LI Mingming

    2005-01-01

    The use of synthetic anabolic steroid methyltestosterone (MT) as growth promoter is prohibited in China. Validations of analytical methods for MT residue in food and the results obtained have become indispensable. The high performance liquid chromatography (HPLC) method for the detection of MT with liquid-liquid extraction by trichloromethanemethanol in carp muscle tissue was preliminarily validated with reference to the following parameters: recovery (accuracy)at the 1, 5 and 10 mgkg-1 level, between-run and within-run CV values (repeatability, also called relative standard deviation(RSD)) and limit of detection. The recoveries were above 80% and the between-run and within-run CV values below 10%for muscle tissue. The limit of detection was 0.05 mgkg-1.

  12. Development of a new ultra-high performance liquid chromatography - tandem mass spectrometry method for determination of ambroxol hydrochloride in serum with pharmacokinetic application

    OpenAIRE

    Vujović Maja M.; Jokanović Milan; Nikolić Goran M.

    2016-01-01

    Ambroxol hydrochloride is an expectorant agent, successfully applied in mucolytic therapy for acute and chronic bronchopulmonary diseases. The drug regulates not only mucus secretion but also showed antioxidant, anti-inflammatory and local anesthetic properties. To supplement the pharmacokinetic and toxicological studies of ambroxol, a rapid ultra-high performance liquid chromatography-tandem mass spectrometry method for the quantitation of ambroxol in rabb...

  13. Determination of cyclodextrins in biological fluids by high-performance liquid chromatography with negative colorimetric detection using post-column complexation with phenolphthalein

    NARCIS (Netherlands)

    Frijlink, H.W.; Visser, J.; Drenth, B.F.H.

    1987-01-01

    A rapid and sensitive high-performance liquid chromatographic method for the analysis of beta- and gamma-cyclodextrin in aqueous biological fluids such as plasma, urine, or tissue homogenate is described. The chromatographic system consists of a microBondapak Phenyl column as stationary phase and a

  14. Development of a new ultra-high performance liquid chromatography - tandem mass spectrometry method for determination of ambroxol hydrochloride in serum with pharmacokinetic application

    OpenAIRE

    Vujović Maja M.; Jokanović Milan; Nikolić Goran M.

    2016-01-01

    Ambroxol hydrochloride is an expectorant agent, successfully applied in mucolytic therapy for acute and chronic bronchopulmonary diseases. The drug regulates not only mucus secretion but also showed antioxidant, anti-inflammatory and local anesthetic properties. To supplement the pharmacokinetic and toxicological studies of ambroxol, a rapid ultra-high performance liquid chromatography-tandem mass spectrometry method for the quantitation of ambroxol in rabb...

  15. Selective on-line detection of boronic acids and derivatives in high-performance liquid chromatography eluates by post-column reaction with alizarin

    NARCIS (Netherlands)

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

    2015-01-01

    An on-line high-performance liquid chromatography (HPLC) method for the rapid and selective detection of boronic acids in complex mixtures was developed. After optimization experiments at an HPLC flow rate of 0.40 mL/min, the HPLC-separated analytes were mixed post-column with a solution of 75 µM al

  16. Analysis of nitroguanidine in Aqueous Solutions by HPLC (High Performance Liquid Chromatography) with electrochemical Detection and Voltammetry

    Science.gov (United States)

    1987-04-01

    The nitroguanidine was analyzed by high performance liquid chromatography (HPLC) with electrochemical detection at a hanging miercury drop electrode...previously reported on the application of solid sorbent collection techniques to the analysis of several explosives in water by high performance liquid chromatography (HPLC

  17. [Separation of zoledronic acid and its related substances by ion-pair reversed-phase high performance liquid chromatography].

    Science.gov (United States)

    Zhang, Xiaoqing; Jiang, Ye; Xu, Zhiru

    2004-07-01

    A rapid and simple ion-pair reversed-phase high performance liquid chromatographic method (HPLC) has been established for the routine analysis of zoledronic acid and its related substances. The chromatographic conditions were optimized based on the satisfactory separation of zoledronic acid from imidazol-1-ylacetic acid, their retention times and peak shape. The excellent separation of zoledronic acid from its related substances, including the remaining imidazol-1-ylacetic acid used in the synthesis of zoledronic acid and other impurities of oxidation and decomposition, was achieved within 9 min on a Hypersil C8 column with UV detection at 220 nm. The mobile phase was a mixture of methanol (20%) and 5 mmo/L phosphate buffer (80%) that contains 6 mmol/L tetrabutylammonium bromide. The resolution factor of zoledronic acid from its adjacent peak was more than 2.5. This is a simple and rapid method for the routine assay of zoledronic acid.

  18. Ion exchange, chromatofocusing and size exclusion high-performance liquid chromatography of the human uterine progesterone receptor.

    Science.gov (United States)

    Holmes, S D; Smith, R G

    1985-12-01

    The human uterine progesterone receptor was subjected to high-performance liquid chromatography on size exclusion, anion exchange and chromatofocusing columns. For the rapid isolation of the receptor, recovery of [3H]progesterone as well as protein from the columns was essential. The size exclusion columns (G2000 SW and G3000 SW) as well as Mono P HR 5/20 chromatofocusing column adsorbed [3H]progesterone and thus were not useful for separation purposes. The anion exchange (polyanion SI-17) and chromatofocusing columns, AX500, and IEX 540 DEAE gave very good recoveries of protein (greater than 90%) and [3H]progesterone; 80, 66 and 88% respectively. These columns gave rapid and reproducible separation of the progesterone receptor from other cytosol proteins.

  19. Ion pair-based dispersive liquid-liquid microextraction followed by high performance liquid chromatography as a new method for determining five folate derivatives in foodstuffs.

    Science.gov (United States)

    Nojavan, Yones; Kamankesh, Marzieh; Shahraz, Farzaneh; Hashemi, Maryam; Mohammadi, Abdorreza

    2015-05-01

    A novel technique for simultaneous determination of five folate derivatives in various food matrices was developed by ion pair-based dispersive liquid-liquid microextraction (IP-DLLME) combined with high-performance liquid chromatography (HPLC). In the proposed method, N-methyl-N,N-dioctyloctan-1-ammonium chloride (aliquat-336) was used as an ion-pair reagent. Effective variables of microextraction process were optimized. Under optimum conditions, the method yielded a linear calibration curve ranging from 1-200 ng g(-1) with correlation coefficients (r(2)) higher than 0.98. The relative standard deviation for the seven analyses was 5.2-7.4%. Enrichment factors for the five folates ranged between 108-135. Limits of detection were 2-4.1 ng g(-1). A comparison of this method with other methods described that the new proposed method is rapid and accurate, and gives very good enrichment factors and detection limits for determining five folate derivatives. The newly developed method was successfully applied for the determination of five folate derivatives in wheat flour, egg yolk and orange juice samples.

  20. Combination of saponification and dispersive liquid-liquid microextraction for the determination of tocopherols and tocotrienols in cereals by reversed-phase high-performance liquid chromatography.

    Science.gov (United States)

    Shammugasamy, Balakrishnan; Ramakrishnan, Yogeshini; Ghazali, Hasanah M; Muhammad, Kharidah

    2013-07-26

    A simple sample preparation technique coupled with reversed-phase high-performance liquid chromatography was developed for the determination of tocopherols and tocotrienols in cereals. The sample preparation procedure involved a small-scale hydrolysis of 0.5g cereal sample by saponification, followed by the extraction and concentration of tocopherols and tocotrienols from saponified extract using dispersive liquid-liquid microextraction (DLLME). Parameters affecting the DLLME performance were optimized to achieve the highest extraction efficiency and the performance of the developed DLLME method was evaluated. Good linearity was observed over the range assayed (0.031-4.0μg/mL) with regression coefficients greater than 0.9989 for all tocopherols and tocotrienols. Limits of detection and enrichment factors ranged from 0.01 to 0.11μg/mL and 50 to 73, respectively. Intra- and inter-day precision were lower than 8.9% and the recoveries were around 85.5-116.6% for all tocopherols and tocotrienols. The developed DLLME method was successfully applied to cereals: rice, barley, oat, wheat, corn and millet. This new sample preparation approach represents an inexpensive, rapid, simple and precise sample cleanup and concentration method for the determination of tocopherols and tocotrienols in cereals.

  1. Simultaneous determination of six synthetic phenolic antioxidants in edible oils using dispersive liquid-liquid microextraction followed by high-performance liquid chromatography with diode array detection.

    Science.gov (United States)

    Xu, Shuangjiao; Liu, Liangliang; Wang, Yanqin; Zhou, Dayun; Kuang, Meng; Fang, Dan; Yang, Weihua; Wei, Shoujun; Xiao, Aiping; Ma, Lei

    2016-08-01

    A simple, rapid, organic-solvent- and sample-saving pretreatment technique, called dispersive liquid-liquid microextraction, was developed for the determination of six synthetic phenolic antioxidants from edible oils before high-performance liquid chromatography with diode array detection. The entire procedure was composed of a two-step microextraction and a centrifugal process and could be finished in about 5 min, only consuming only 25 mg of sample and 1 mL of the organic solvent for each extraction. The influences of several important parameters on the microextraction efficiency were thoroughly investigated. Recovery assays for oil samples were spiked at three concentration levels, 50, 100 and 200 mg/kg, and provided recoveries in the 86.3-102.5% range with a relative standard deviation below 3.5%. The intra-day and inter-day precisions for the analysis were less than 3.8%. The proposed method was successfully applied for the determination of synthetic phenolic antioxidants in different oil samples, and satisfactory results were obtained. Thus, the developed method represents a viable alternative for the quality control of synthetic phenolic antioxidant concentrations in edible oils.

  2. High-Performance Liquid Chromatography Determination of Meloxicam and Piroxicam with Ultraviolet Detection

    Directory of Open Access Journals (Sweden)

    Sherry Cox

    2014-01-01

    Full Text Available A simple accurate and sensitive high-performance liquid chromatographic method for the determination of meloxicam and piroxicam concentrations in small volume plasma samples has been developed. Following a liquid extraction using chloroform, samples were separated by reversed-phase high-performance liquid chromatography on an XBridge C18 column (4.6 × 250 mm and quantified using ultraviolet detection at 360 nm. The mobile phase was a mixture of water with glacial acetic acid (pH 3.0 and acetonitrile (50 : 50, with a flow rate of 1.0 mL/min. The standard curve ranged from 5 to 10,000 ng/mL for meloxicam in bearded dragon (Pogona vitticeps plasma and piroxicam in crane (Grus rubicunda plasma. Intra- and interassay variability for meloxicam and piroxicam were less than 10% and the average recovery was greater than 90% for both drugs. This method was developed in bearded dragon and crane plasma and should be applicable to any species, making it useful for those investigators dealing with small sample volumes, particularly when conducting pharmacokinetics studies which require multiple sampling from the same animal.

  3. Analysis of some selected catechins and caffeine in green tea by high performance liquid chromatography.

    Science.gov (United States)

    El-Shahawi, M S; Hamza, A; Bahaffi, S O; Al-Sibaai, A A; Abduljabbar, T N

    2012-10-15

    Green tea seems to have a positive impact on health due to the catechins-found as flavanols. Thus, the present study was aimed to develop a low cost reversed phase high performance liquid chromatographic (HPLC) method for simultaneous determination of flavanol contents, namely catechin (C), epicatechin (EC), epigallocatechin (EGC), epicatechin 3-gallate (ECG) and epigallocatechin 3-gallate (EGCG) and caffeine in 29 commercial green tea samples available in a Saudi Arabian local market. A C-18 reversed-phase column, acetonitrile-trifluoroacetic acid as a mobile phase, coupled with UV detector at 205 nm, was successfully used for precise analysis of the tested analytes in boiled water of digested tea leaves. The average values of N (No. of theoretical plates), HETP (height equivalent of theoretical plates) and R(s) (separation factor) (at 10 μg ml(-1) of the catechins EC, EGC, EGCG and ECG) were 2.6×10(3)±1.2×10(3), 1.7×10(-3)±4.7×10(-4) cm and 1.7±5.53×10(-2), respectively. The developed HPLC method demonstrated excellent performance, with low limits of detection (LOD) and quantification (LOQ) of the tested catechins of 0.004-0.05 μg ml(-1) and 0.01-0.17 μg ml(-1), respectively, and recovery percentages of 96-101%. The influence of infusion time (5-30 min) and temperature on the content of the flavanols was investigated by HPLC. After a 5 min infusion of the tea leaves, the average concentrations of caffeine, catechin, EC, EGC, ECG and EGCG were found to be in the ranges 0.086-2.23, 0.113-2.94, 0.58-10.22, 0.19-24.9, 0.22-13.9 and 1.01-43.3 mg g(-1), respectively. The contents of caffeine and catechins followed the sequence: EGCG>EGC>ECG>EC>C>caffeine. The method was applied satisfactorily for the analysis of (+)-catechin, even at trace and ultra trace concentrations of catechins. The method was rapid, accurate, reproducible and ideal for routine analysis.

  4. Analysis of sup 99m Tc-labeled radiopharmaceuticals by high-performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Muto, Toshio (Tokyo Metropolitan Isotope Research Center (Japan))

    1990-02-01

    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 {sup 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 {sup 99m}Tc-HSA and the other {sup 99m}Tc-agents, respectively. The analysis of low molecular weight {sup 99m}Tc-agents (e.g. {sup 99m}Tc-DTPA, {sup 99m}Tc-DMSA, {sup 99m}Tc-pyrophosphate, {sup 99m}Tc-phytic acid, {sup 99m}Tc-MDS, {sup 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 {sup 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 {sup 99m}TcO{sub 4} was observed in {sup 99m}Tc-pyrophosphate, {sup 99m}Tc-phytic acid, {sup 99m}Tc-MDP, {sup 99m}Tc-HMDP, and impurities of {sup 99m}Tc-labeled species and {sup 99m}TcO{sub 4}, were observed in {sup 99m}Tc-HIDA, {sup 99m}Tc-HIDA, {sup 99m}Tc-HSA. The radiochemical impurities of the {sup 99m}Tc-radiopharmaceuticals were ranged between 90 and 100 %. From these results, radio-HPLC has been shown to be suitable method for analysis of {sup 99m}Tc-radiopharmaceuticals, with rapidity and excellent precision. (author).

  5. 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.

  6. On-line radiochemical assay for monoamine oxidase utilizing high-performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Nissinen, E.; Linko-Loeppoenen SMae; Maennistoe P4

    1984-12-01

    A fast and sensitive assay for the determination of monoamine oxidase activity was developed. The method is based on the separation and quantitation of /sup 14/C-labeled assay products by high-performance liquid chromatography, which is interfaced directly into a flow-through radioactivity detector. This allows on-line quantitation of the radioactive compounds with picomole sensitivity. The method makes possible the complete separation and detection of the deaminated products of monoamine oxidase A and B substrates benzylamine and 5-hydroxytryptamine, respectively. This assay has been applied to the measurement of monoamine oxidase A and B activities in rat brain.

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

    DEFF Research Database (Denmark)

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

    1993-01-01

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

  8. Determination of Constituents in Hair Dyes by Reversed-phase High-Performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    A reversed-phase high-performance liquid chromatographic method for the separation and quantitation of 1,4-phenylenediamine, 4-aminophenol, 3-aminophenol, 1,3-benzendiol and 2,6-phenylenediamine in hair dyes was developed. The separation was carried out on a μ-Bondapak C18 column with methanol-water containing 0.1% triethylamine and 0.02 mol/L NH4Ac (pH=5.20) (10:90, vol./vol.) as the mobile phase. Two real samples were analyzed.

  9. Determination of enantiomeric impurity of etomidate by high performance liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Objective To determine enantiomeric impurity of etomidate using high performance liquid chromatography. Methods (R)-etomidate and (S)-etomidate were separated on a CHIRALPAK AD-H column. The mobile phase consisted of 20∶80(v/v) isopropanol-n-hexane. The flow rate of the mobile phase was 0.5mL/min. The detected wavelength was 242nm. Results (R)-etomidate and (S)-etomidate could be separated completely under these conditions. The precision of (R)-etomidate was 1.57% (n=3). The limit of detection of (R)-etomid...

  10. Anomalous properties of flavonoids in reversed phase high performance liquid chromatography

    Science.gov (United States)

    Zenkevich, I. G.; Gushchina, S. V.

    2011-09-01

    It is shown through reversed phase high performance liquid chromatography that a characteristic feature of such abundant natural flavonoids as flavon-3-ols is an anomalously strong antibate dependence of their retention indices ( RI) on the organic solvent concentration ( C) in the eluent, dRI/ dC < 0. In order to interpret this anomaly, the specific optical rotation values [α]{D/20} of natural (+)-(2 R,3 R)-dihydroquercetin in different solvents are compared, confirming the reverse formation of hydrated flavonoids in aqueous solutions.

  11. Study of Saiga Horn Using High-Performance Liquid Chromatography with Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Kateřina Mikulíková

    2012-01-01

    Full Text Available The saiga horns have been investigated the using of modern analytic methods. High-performance liquid chromatography (HPLC with mass-spectrometric (MS and MS/MS detection and polyacrylamide gel electrophoresis (PAGE were used. It could be concluded that basic proteins of the saiga horns are keratins and collagen. The basic representation protein in all samples is keratin type I microfibrillar (from sheep, keratin type II microfibrillar (from sheep, collagen type I (α1 (from bovine and collagen type I (α2 (from bovine. Free amino acids we determined in all samples are nontreated by enzyme.

  12. Determination of cortisol and cortisone in urine using high-performance liquid chromatography with UV detection.

    Science.gov (United States)

    Jenner, D A; Richards, J

    1985-01-01

    The application of reversed-phase gradient high-performance liquid chromatography with UV detection to the determination of cortisol and cortisone in 24-h urine samples is described. The method employs Sep-pak C18 cartridges for the part-purification and concentration of the corticosteroids, with sample enrichment at the head of an HPLC pre-column and separation using water/acetonitrile gradient. The internal standard is 6alpha-methylprednisolone. Measurement of both cortisone and cortisol provides further information on adrenocortical function. 24-hour excretion rate data from normal subjects are reported.

  13. Determination of rhoifolin and daidzin in human plasma by high-performance liquid chromatography.

    Science.gov (United States)

    Ishii, K; Urano, S; Furuta, T; Kasuya, Y

    1994-05-13

    A method for determining flavonoids in human plasma is presented for application to pharmacokinetic studies of two flavonoids, rhoifolin and daidzin. Isocratic reversed-phase high-performance liquid chromatography (HPLC) was used with genistin as an internal standard and solid-phase extraction using a Sep-Pak C18 cartridge. The mobile phases were acetonitrile-0.1 M ammonium acetate solution (20:80, v/v) for rhoifolin and methanol-0.1 M ammonium acetate solution (33:67, v/v) for daidzin. The detection limits on-column were 2 ng for rhoifolin and 0.5 ng for daidzin.

  14. Determination of contents of 10-Hydroxycamptothecin in Camptotheca acuminata by high-performance liquid chromatogram

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The determination method of 10-hydroxycamptothecin in Camptotheca acuminata fruits by high-performance liquid chromatogram (HPLC) was studied. The HPLC analysis was performed on a HIQ sil C18(4.6×250 mm) column with mobile phase of acetonitrile-water (3:7, V:V), flow rate 1 mL@(min-1 and UV detective wavelength 266 nm. Extracting 10-hydroxycamptothecin by ultrasonic method from fruits of C. acuminata to prepare samples for analysis was systematically discussed. The optimal extraction condition was carried out by 60% alcohol solution at 60℃ for 50 minutes.

  15. Fluorimetric assay for ornithine decarboxylase by high-performance liquid chromatography.

    Science.gov (United States)

    Haraguchi, K; Kai, M; Kohashi, K; Ohkura, Y

    1980-12-05

    A highly sensitive method for the assay of ornithine decarboxylase in sample solutions prepared from rat tissue homogenate is described which employs high-performance liquid chromatography with fluorescence detection. Putrescine formed from ornithine under the optimal conditions for the enzyme reaction is treated by Cellex P column chromatography for clean-up and converted into the fluorescamine derivative in the presence of cupric ion which inhibits the reaction of interfering amines with fluorescamine. The derivative is separated by reversed-phase chromatography on LiChrosorb RP-18 with linear gradient elution. The lower limit of detection for putrescine formed enzymatically is 5 pmol.

  16. Detection of cheese whey and caseinomacropeptide in fermented milk beverages using high performance liquid chromatography

    OpenAIRE

    E.H.P. Andrade; M.R. Souza; Fonseca,L.M.; C.F.A.M. Penna; M.M.O.P. Cerqueira; T. Roza; Seridan,B.; M.F.S. Resende; Pinto, F. A.; C.N.B.C. Villanoeva; Leite,M.O.

    2014-01-01

    Cheese whey level and caseinomacropeptide (CMP) index of fermented milk beverages added with four levels of cheese whey (0, 10, 20, and 40%) and stored at 8-10oC for 0, 7, 14 and 21 days were determined by high performance liquid chromatography-gel filtration (HPLC-GF). Additionally, the interference of the starter culture and the storage time on the detection of cheese whey and CMP were investigated. Refrigerated storage up to 21 days did not affect (P>0.05) cheese whey and CMP amounts in mi...

  17. Simultaneous determination of pseudoephedrine hydrochloride and cetrizine hydrochloride by reverse phase high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Nalini C

    2006-01-01

    Full Text Available A reversed phase high performance liquid chromatographic method has been developed using Shimadzu HPLC-VP series, LC-10 ATV pump, SPD10 AVP and C8 column, for simultaneous determination of pseudoephedrine hydrochloride and cetrizine hydrochloride in three marketed tablet formulations (extended release. The mobile phase consists of phosphate buffer of pH 7.0 and acetonitrile HPLC grade in the ratio of 1:1. The flow rate was maintained at 1 ml/min and the ultraviolet detection was done at 242 nm, which is the isosbestic point. Linearity coefficients, assay values, recovery studies and repeatability studies showed that the method is accurate and precise.

  18. Determination of nadolol in serum by high-performance liquid chromatography with fluorimetric detection.

    Science.gov (United States)

    Noguchi, H; Yoshida, K; Murano, M; Naruto, S

    1992-01-17

    A sensitive high-performance liquid chromatographic method for a routine assay of nadolol in serum is described. Serum samples spiked with atenolol (internal standard) were extracted with diethyl ether. After centrifugation, the organic layer was evaporated to dryness. The residue was redissolved in the mobile phase and injected onto an octadecyl silica column (150 mm x 4.6 mm I.D.). The mobile phase was 0.05 M ammonium acetate (pH 4.5)-acetonitrile (85:15, v/v). Fluorometric detection (excitation 230 nm, emission 300 nm) was used. The minimum detectable level of nadolol in serum was 1 ng/ml.

  19. A new method for high-performance liquid chromatographic determination of drotaverine in plasma.

    Science.gov (United States)

    Mezei, J; Küttel, S; Szentmiklósi, P; Marton, S; Rácz, I

    1984-10-01

    A sensitive, specific high-performance liquid chromatographic procedure was developed for the determination of plasma drotaverine levels. Basic plasma samples were adjusted to pH 1.5 and extracted with chloroform. HPLC [n-heptane-dichloromethane-diethylamine (50:25:2)] on a microporous silica column, with a variable-wavelength UV detector set at 302 nm allowed the measurement of drotaverine at the 50-ng/mL level. The utility of this method for determination of drotaverine in dog and rat plasma was demonstrated.

  20. Separation of polyamines, conjugated to DNA, by reversed-phase high-performance liquid chromatography.

    Science.gov (United States)

    Mateos, J L; Reyes, A; Vicente, C; Legaz, M E

    2000-02-18

    Genomic DNA was isolated from the lichen Evernia prunastri in order to analyze by high-performance liquid chromatography the occurrence of polyamines conjugated to the macromolecule. The acid-insoluble (PH) fraction of this DNA contained mainly conjugated spermidine, although small amounts of free putrescine and spermidine were also present. The PH fraction of DNA also contained conjugated evernic acid, the main phenol produced by this lichen species. Conjugation of polyamines to calf thymus DNA was carried out under in vitro conditions. Conjugation was to spermidine and mainly to spermine and produced DNA compactation. Evernic acid enhanced the action of polyamines in order to produce DNA aggregation.

  1. Studies on the Fingerprint of Yubaizhi by High-Performance Liquid Chromato Graphy

    Institute of Scientific and Technical Information of China (English)

    YU Yu-hong; SUN Yan-ke; LI Xue-chun; DAI Rong-ji; MENG Wei-wei; DENG Yu-lin

    2008-01-01

    The investigation of fingerprint of Yubaizhi was performed by high-performance liquid chromatography (HPLC). Chromatographic conditions were: the mobile phase was the mixture of acetonitrile-water (43:57, v/v); the flow rate was 1mL/min; the column temperature was 40℃; the detection wavelength was 254nm. The stability, precision and reproducibility were performed according to "Specification of Fingerprint of Injection of Chinese Traditional Medicine (temporarily)" regulated by State Food and Drug Administration(SFDA). The HPLC fingerprints of Yubaizhi from various locations were well constant.

  2. 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.

  3. 固相萃取和高效液相色谱相结合快速测定苦瓜甙A的含量%Rapid Analysis of Momordicoside A in Bitter Melon by High Performance Liquid Chromatography Following Solid Phase Extraction

    Institute of Scientific and Technical Information of China (English)

    王水; 唐琳; 郭亦然; 颜钫; 陈放

    2001-01-01

    建立了一个快速、简单、准确的固相萃取和高效液相色谱相结合的测定苦瓜甙A含量的方法。样品经石墨碳固相萃取管(3 mL/250 mg)纯化后以高效液相色谱检测。色谱柱为C18,流动相为V(乙腈)∶V(甲醇)∶V(50 mmol/L磷酸二氢钾缓冲液)=25∶20∶60,流速为0.8 mL/min,检测波长为208 nm。标准曲线自10 mg/L到1*!000 mg/L呈线形关系(r2=0.999*!2)。该方法具有很好的重现性,日内或日间的相对标准偏差和相对平均误差均小于10%。样品回收率大于90%。%A rapid, simple and accurate method for the determination of momordicoside A has been established using solid phase extraction (SPE) followed by high performance liquid chromatography (HPLC). Sample was processed by SPE on a Carb cartridge (3 mL/250 mg), and was then determined by HPLC on a C18 column (4.6 mm i.d.×250 mm, 5 μm) with V(acetonitrile)∶V(methanol)∶V(50 mmol/L potassium dihydrogen phosphate buffer)=25∶20∶60 as mobile phase (0.8 mL/min) and UV detection at 208 nm. The calibration curves were linear from 10 mg/L to 1*!000 mg/L (r2=0.999 2). The analytical method was shown to be highly reproducible, giving all of the relative standard deviations and relative mean errors less than 10% for both intra-day and inter-day determinations. The absolute recoveries were greater than 90%.

  4. 变性高效液相色谱技术快速检测下呼吸道致病性细菌的实验研究%Experimental study on rapid detection of lower respiratory tract pathogenic bacteria by denaturing high-performance liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    张晓燕; 刘朝晖; 梁志科; 王汉平; 叶惠芬; 杨银梅; 谢健晋

    2008-01-01

    目的 运用变性高效液相色谱(DHPLC)技术检测5种下呼吸道致病性细菌,建立一种快速检测下呼吸道致病性细菌的分子生物学方法.方法 以下呼吸道致病性细菌16S rRNA基因的保守区设计通用引物,并在引物前加上40-bpGC,特异性扩增该基因的保守区和可变区,运用DH-PLC技术对PCR产物进行分析.选取50株临床分离菌株验证该方法的有效性.结果 通用引物可特异性扩增细菌16S rRNA,PCR产物经DHPLC分析后每种细菌均能得到特征性的洗脱峰.DHPLC检测临床分离菌株结果显示,与常规培养方法的符合率为100%.结论 DHPLC技术具有准确、简便、快捷和高通量等特点,在临床检测下呼吸道致病性细菌中具有潜在的应用价值.%Objective To construct a new molecular biological method for the analysis of microbial species in lower respiratory tract infections based on 16S rRNA gene by denaturing high-performance liquid chromatograph(DHPLC).Methods The universal primer set was analyzed basing on the highly conserved regions of 16S rRNA gene.DNA amplicons of lower respiratory tract were analyzed by DHPLC to generate peak profiles respectively.The incorporation of 40-bpGC clamp into the amplification primet was essential to effectively discriminate genetic differences identification.Results The primers could only amplify bacterial 16S rRNA.Bacterial of amplicons which incorporation of a 40-bpGC clamp were effectively discriminated genetic differences in DHPLC.The results of clinical isolares identification showed 100%according with the traditional method.Conclusion DHPLC has not only high accuracy,but also is a convenient,rapid and high-through technique for the discrimination bacteria.It has potential value in the detection of lower respiratory pathogenic bacteria.

  5. Rapid determination of three Pacific ciguatoxins in muscle of moral eel by high performance liquid chromatography-tandem mass spectrometry%高效液相色谱-串联质谱法快速测定海鳗鱼肉中3种太平洋雪卡毒素

    Institute of Scientific and Technical Information of China (English)

    麦艳玲; 林珊珊; 肖陈贵; 岳振峰; 吴佳俊; 康海宁; 林群声; 陈荔

    2014-01-01

    目的:建立了一种平行振荡方法萃取海鳗鱼肉中的3种太平洋雪卡毒素(P-CTX-1、P-CTX-2、P-CTX-3),并采用高效液相色谱-电喷雾串联质谱法同时测定P-CTX-1、P-CTX-2、P-CTX-3残留量的分析方法。方法海鳗等样品经甲醇并采用平行振荡方法提取, C18固相萃取柱净化后,采用高效液相色谱-电喷雾串联质谱分析测定。结果以基质标准曲线外标法定量,线性范围为0.01∼50.0 ng/g,相关系数均大于0.999(r≥0.999, n=3), P-CTX-1、P-CTX-2、P-CTX-3的定量限分别为(LOQs,S/N=10)0.009、0.019、0.017 ng/g。以0.05、0.10、0.50 ng/g 3个添加浓度水平进行方法验证,回收率分别为72.9~81.0%、63.0~77.9%、46.3~69.5%,相对标准偏差分别为2.3~2.7%、2.1~12.2%、6.1~15.3%。结论该方法简便快速、灵敏度高,适用于鱼肉中同时快速检测3种太平洋雪卡毒素。%Objective A parallel shaking method was developed to extract three Pacific ciguatoxins (P-CTX-1, P-CTX-2, P-CTX-3) in muscle samples of moray eel and a rapid simultaneous quantification me-thod was optimized to quantify P-CTXs in fish muscle using high performance liquid chromatography coupled with electrospray-tandem mass spectrometry (HPLC-MS/MS).MethodsMuscle samples of moray eel were extracted in methanol using parallel shaker. Crude extract was then cleaned-up by C18 solid-phase extraction (SPE) cartridges. High performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was employed to identify and quantify P-CTXs.ResultsThe quantification of P-CTX-1, P-CTX-2 and P-CTX-3 were performed using matrix-matched calibrations with correlation coefficients above 0.999 (r≥0.999,n=3). The linearity of the calibration curves ranged from 0.01~50.0 ng/g. The limits of quantification (LOQs,S/N=10) of P-CTX-1, P-CTX-2 and P-CTX-3 were 0.009, 0.019, and 0.017 ng/g. Method validation was carried out by spiking three levels of P-CTXs (i.e. 0.05, 0.10, and 0.50 ng/g) to fish muscle. The

  6. Gold nanoparticle decorated graphene oxide/silica composite stationary phase for high-performance liquid chromatography.

    Science.gov (United States)

    Liang, Xiaojing; Wang, Xusheng; Ren, Haixia; Jiang, Shengxiang; Wang, Licheng; Liu, Shujuan

    2014-06-01

    In the initial phase of this study, graphene oxide (GO)/silica was fabricated by assembling GO onto the silica particles, and then gold nanoparticles (GNPs) were used to modify the GO/silica to prepare a novel stationary phase for high-performance liquid chromatography. The new stationary phase could be used in both reversed-phase chromatography and hydrophilic interaction liquid chromatography modes. Good separations of alkylbenzenes, isomerides, amino acids, nucleosides, and nucleobases were achieved in both modes. Compared with the GO/silica phase and GNPs/silica phase, it is found that except for hydrophilicity, large π-electron systems, hydrophobicity, and coordination functions, this new stationary phase also exhibited special separation performance due to the combination of 2D GO with zero-dimensional GNPs.

  7. Fractionation of Gibberellins in plant extracts by reverse phase high performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Jones, M.G.; Metzger, J.D.; Zeevaart, J.A.D.

    1980-02-01

    In studies on endogenous plant gibberellins (GAs), reverse phase (Bondapak C/sub 18/) high performance liquid chromatography (HPLC) has proved to be a useful method for the fractionation of plant extracts. The behavior of 18 authentic GAs in such a chromatographic system is described. The main factors determining chromatographic behavior are the degree and the position of hydroxylation of the GA. As an illustration of the use of reverse phase HPLC, the endogeneous GAs of immature seeds of Pharbitis nil L., strain Violet, were reinvestigated. The presence of gibberellins A/sub 3/, A/sub 5/, A/sub 17/, A/sub 20/, and A/sub 29/ was confirmed by gas-liquid chromatography-mass spectrometry. In addition, two other GAs, A/sub 19/ and A/sub 44/, were also identified in extracts of this material.

  8. High-performance liquid-catalyst fuel cell for direct biomass-into-electricity conversion.

    Science.gov (United States)

    Liu, Wei; Mu, Wei; Deng, Yulin

    2014-12-01

    Herein, we report high-performance fuel cells that are catalyzed solely by polyoxometalate (POM) solution without any solid metal or metal oxide. The novel design of the liquid-catalyst fuel cells (LCFC) changes the traditional gas-solid-surface heterogeneous reactions to liquid-catalysis reactions. With this design, raw biomasses, such as cellulose, starch, and even grass or wood powders can be directly converted into electricity. The power densities of the fuel cell with switchgrass (dry powder) and bush allamanda (freshly collected) are 44 mW cm(-2) and 51 mW cm(-2) respectively. For the cellulose-based biomass fuel cell, the power density is almost 3000 times higher than that of cellulose-based microbial fuel cells. Unlike noble-metal catalysts, POMs are tolerant to most organic and inorganic contaminants. Therefore, almost any raw biomass can be used directly to produce electricity without prior purification.

  9. An electrospray ionization-ion mobility spectrometer as detector for high- performance liquid chromatography.

    Science.gov (United States)

    Zühlke, Martin; Riebe, Daniel; Beitz, Toralf; Löhmannsröben, Hans-Gerd; Zenichowski, Karl; Diener, Marc; Linscheid, Michael W

    2015-01-01

    The application of electrospray ionization (ESI) ion mobility (IM) spectrometry on the detection end of a high-performance liquid chromatograph has been a subject of study for some time. So far, this method has been limited to low flow rates or has required splitting of the liquid flow. This work presents a novel concept of an ESI source facilitating the stable operation of the spectrometer at flow rates between 10 μL mn(-1) and 1500 μL min(-1) without flow splitting, advancing the T-cylinder design developed by Kurnin and co-workers. Flow rates eight times faster than previously reported were achieved because of a more efficient dispersion of the liquid at increased electrospray voltages combined with nebulization by a sheath gas. Imaging revealed the spray operation to be in a rotationally symmetric multijet mode. The novel ESI-IM spectrometer tolerates high water contents (≤90%) and electrolyte concentrations up to 10mM, meeting another condition required of high-performance liquid chromatography (HPLC) detectors. Limits of detection of 50 nM for promazine in the positive mode and 1 μM for 1,3-dinitrobenzene in the negative mode were established. Three mixtures of reduced complexity (five surfactants, four neuroleptics, and two isomers) were separated in the millisecond regime in stand-alone operation of the spectrometer. Separations of two more complex mixtures (five neuroleptics and 13 pesticides) demonstrate the application of the spectrometer as an HPLC detector. The examples illustrate the advantages of the spectrometer over the established diode array detector, in terms of additional IM separation of substances not fully separated in the retention time domain as well as identification of substances based on their characteristic Ims.

  10. THE USE OF HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY AS SCREENING TECHNIQUE FOR PECTIN AND PECTIN SUBSTANCES OF DIETARY FIBERS

    Directory of Open Access Journals (Sweden)

    G.N. Ignatyeva

    2015-02-01

    Full Text Available The peak of the unspecified compound of quantitative analysis of organic acids in biological materials by using high-performance liquid chromatography was identified. This peak corresponded to polygalacturonic acid. A High-performance liquid chromatographic method was developed to identify and quantify of water soluble pectin fraction and insoluble pectin fraction, galacturonic acid and polygalacturonic acid. The method based on the extraction of free water soluble fraction with phosphate buffer (pH 4.8 or with 0.01 N phosphoric acid and on the acid hydrolysis of insoluble pectin fraction in presence 0.1 N HNO3. The pectin fractions were separated and determined on a C610H column using elution with 0.01 N phosphoric acids and ultraviolet detection at 210 nm. Galacturonic acid and polygalacturonic acid were used as internal standard. The calibration curves were presented for each of the standards. The determinations were performed in the linear range of 0.04÷0.17% w/v for both pectin fractions. Proposed procedure can considered as a rapid method for quantitative pectin determination in the food, food nutrition’s food, and pharmaceutical agents.

  11. Simultaneous determination of four marker components in Yukmijihwang Tang by high performance liquid chromatography/diode array detector.

    Science.gov (United States)

    Won, Jin Bae; Ma, Jin Yeul; Ma, Choong Je

    2010-04-01

    Simultaneous determination method of four marker components, paeoniflorin, loganin, 5-hydroxymethyl-2-furaldehyde and paeonol in traditional herbal medicine, Yukmijihwang tang, was developed to achieve rapidly and systematic quality control by high performance liquid chromatography coupled with diode array detection. To develop this high performance liquid chromatography method, C(18) column (5 microm, 120 A, 4.6 mm x 150 mm) was used with gradient elution of water and methanol as mobile phase. Validation of the chromatography method was evaluated by linearity, recovery, and precision test. Calibration curves of standard components showed excellent linearity with R(2) > 0.9990. Limits of detection and limits of quantification were in the ranges 0.01-0.02 and 0.04-0.07 microg/mL, respectively. The relative standard deviations of data of the intra-day and inter-day experiments were less than 1.15% and 3.76%, respectively. The results of recovery test were found to range from 94.88 to 107.43% with relative standard deviation values 0.07-2.66%. The results of validation suggested that this method was very accurate and stabilized.

  12. Feeding of liquid silicon for high performance multicrystalline silicon with increased ingot height and homogenized resistivity

    Science.gov (United States)

    Krenckel, Patricia; Riepe, Stephan; Schindler, Florian; Strauch, Theresa

    2017-04-01

    Feeding of liquid silicon during the directional solidification process is a promising opportunity for cost reduction by increased throughput and improved material homogeneity due to constant resistivity over ingot height. In this work, a liquid feeding apparatus was developed for an industrial type directional solidification furnace. One n-type G2 sized High Performance multicrystalline ingot with liquid feeding of additional 14 kg of undoped silicon feedstock was crystallized. The resistivity was kept within a range of ±0.1 Ω cm of the target resistivity during the feeding sequence. A smaller mean grain area growth was observed during feeding, whereas the area fraction of recombination active dislocation structures was as low as in a reference ingot. Increased interstitial oxygen and substitutional carbon concentrations were measured for the ingot with liquid feeding. The measured mean bulk lifetime of 190 μs for passivated wafers in the feeding sequence can probably be increased by further pre-melting crucible improvements. For this laboratory experiment, energy reductions of 2% per wafer and time savings of 16% per wafer were realized.

  13. Simultaneous determination of airborne carbamates in workplace by high performance liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Li, Hong-Ping; Li, Jen-Hou; Li, Gwo-Chen; Jen, Jen-Fon

    2004-06-17

    A high performance liquid chromatography with fluorescence (HPLC-F) detector was examined to simultaneous determination of airborne carbamates in the workplace of manufactory. The OVS-2 air sampling tube filled with glass fiber filter or quartz fiber and combined filter/XAD-2 were evaluated to collect nine commonly used carbamates (Carbofuran, Isoprocarb, Methomyl, Metolcarb, Thiodicarb, Carbaryl, Oxamyl, Methiocarb, and Prpoxur) from the air of manufactory in high humidity country. After being extracted with acetonitrile from sampling tubes, the carbamates were determined by high performance liquid chromatography with fluorescence detection posterior to on-line derivatization. The collection of carbamates and the recovery of extraction from glass wool fiber in several concentration levels, and from quartz filter were evaluated. The storage stability of carbamates was also tested. Results indicated that the HPLC-fluorescence method offers satisfactory resolution and sensitivity in carbamate analysis. With the glass fiber filter and combined filter/XAD-2, the Carbofuran, Isoprocarb, Methomyl, Metolcarb, and Thiodicarb were stable for a 28-day storage test, Carbaryl and Oxamyl for 14 days, and Methiocarb and Prpoxur for 7 days. All of these pesticides were with detection limit of 3mugm(-3). It is suited for environmental monitoring. The airborne carbamates in different areas of the manufactory were measured.

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

    Science.gov (United States)

    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 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. PMID:26989559

  15. 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.

  16. Determination of Niacinamide in Lotions and Creams Using Liquid-Liquid Extraction and High-Performance Liquid Chromatography

    Science.gov (United States)

    Usher, Karyn M.; Simmons, Carolyn R.; Keating, Daniel W.; Rossi, Henry F., III

    2015-01-01

    Chemical separations are an important part of an undergraduate chemistry curriculum. Sophomore students often get experience with liquid-liquid extraction in organic chemistry classes, but liquid-liquid extraction is not as often introduced as a quantitative sample preparation method in honors general chemistry or quantitative analysis classes.…

  17. Determination of Niacinamide in Lotions and Creams Using Liquid-Liquid Extraction and High-Performance Liquid Chromatography

    Science.gov (United States)

    Usher, Karyn M.; Simmons, Carolyn R.; Keating, Daniel W.; Rossi, Henry F., III

    2015-01-01

    Chemical separations are an important part of an undergraduate chemistry curriculum. Sophomore students often get experience with liquid-liquid extraction in organic chemistry classes, but liquid-liquid extraction is not as often introduced as a quantitative sample preparation method in honors general chemistry or quantitative analysis classes.…

  18. Synthesis of Pt3Ni Microspheres with High Performance for Rapid Degradation of Organic Dyes

    Science.gov (United States)

    Wang, Min; Yang, Yushi; Long, Jia; Mao, Zhou; Qiu, Tong; Wu, Qingzhi; Chen, Xiaohui

    2015-05-01

    In this study, Pt3Ni microspheres consisted of nanoparticles were synthesized without addition of surfactants via the solvothermal route. The obtained sample was characterized by X-ray diffraction (XRD), inductively coupled plasma-atomic emission spectrometer (ICP-AES), X-ray photoelectron spectroscopy (XPS), and field-emission scanning electron microscopy (FESEM). Furthermore, the catalytic performance of as-synthesized Pt3Ni microspheres was evaluated on the degradation of different organic dyes (methylene blue, methyl orange, Congo red, and rhodamine B). The results show that different dyes were rapidly decomposed by Pt3Ni microspheres in different pathways. Among different dyes, the formation and further degradation of the intermediates was observed during the degradation of methylene blue and methyl orange, suggesting the indirect degradation process of these dyes. This study provides not only a promising catalyst for the removal of organic contaminants for environment remediation, but also new insights for Pt3Ni alloy as a high-performance catalyst in organic synthesis.

  19. Synthesis of Pt3Ni microspheres with high performance for rapid degradation of organic dyes.

    Science.gov (United States)

    Wang, Min; Yang, Yushi; Long, Jia; Mao, Zhou; Qiu, Tong; Wu, Qingzhi; Chen, Xiaohui

    2015-12-01

    In this study, Pt3Ni microspheres consisted of nanoparticles were synthesized without addition of surfactants via the solvothermal route. The obtained sample was characterized by X-ray diffraction (XRD), inductively coupled plasma-atomic emission spectrometer (ICP-AES), X-ray photoelectron spectroscopy (XPS), and field-emission scanning electron microscopy (FESEM). Furthermore, the catalytic performance of as-synthesized Pt3Ni microspheres was evaluated on the degradation of different organic dyes (methylene blue, methyl orange, Congo red, and rhodamine B). The results show that different dyes were rapidly decomposed by Pt3Ni microspheres in different pathways. Among different dyes, the formation and further degradation of the intermediates was observed during the degradation of methylene blue and methyl orange, suggesting the indirect degradation process of these dyes. This study provides not only a promising catalyst for the removal of organic contaminants for environment remediation, but also new insights for Pt3Ni alloy as a high-performance catalyst in organic synthesis.

  20. Space-to-Space Power Beaming Enabling High Performance Rapid Geocentric Orbit Transfer

    Science.gov (United States)

    Dankanich, John W.; Vassallo, Corinne; Tadge, Megan

    2015-01-01

    The use of electric propulsion is more prevalent than ever, with industry pursuing all electric orbit transfers. Electric propulsion provides high mass utilization through efficient propellant transfer. However, the transfer times become detrimental as the delta V transitions from near-impulsive to low-thrust. Increasing power and therefore thrust has diminishing returns as the increasing mass of the power system limits the potential acceleration of the spacecraft. By using space-to-space power beaming, the power system can be decoupled from the spacecraft and allow significantly higher spacecraft alpha (W/kg) and therefore enable significantly higher accelerations while maintaining high performance. This project assesses the efficacy of space-to-space power beaming to enable rapid orbit transfer while maintaining high mass utilization. Concept assessment requires integrated techniques for low-thrust orbit transfer steering laws, efficient large-scale rectenna systems, and satellite constellation configuration optimization. This project includes the development of an integrated tool with implementation of IPOPT, Q-Law, and power-beaming models. The results highlight the viability of the concept, limits and paths to infusion, and comparison to state-of-the-art capabilities. The results indicate the viability of power beaming for what may be the only approach for achieving the desired transit times with high specific impulse.

  1. Ultrasound-assisted ionic liquid-based homogeneous liquid-liquid microextraction high-performance liquid chromatography for determination of tanshinones in Salvia miltiorrhiza Bge. root.

    Science.gov (United States)

    Wang, Zhibing; Cao, Bocheng; Yu, Aimin; Zhang, Hanqi; Qiu, Fangping

    2015-02-01

    The ultrasound-assisted ionic liquid-based homogeneous liquid-liquid microextraction has been developed and applied to the extraction of four tanshinones, including dihydrotanshinone, tanshinone I, cryptotanshinone and tanshinone IIA in Salvia miltiorrhiza Bge. root. High performance liquid chromatography was applied to the separation and determination of the analytes. The ionic liquid was used as extraction solvent and target analytes were extracted with help of ultrasound. Then, ion-pairing agent was added into the sample solution, which resulted in the formation of water-insoluble ionic liquid in the solution. The phase separation was performed by centrifugation. The extraction, concentration and purification of target analytes were performed simultaneously. The experimental parameters, including type and volume of ionic liquid, sample amount, the size of sample particle, pH value of extraction medium, extraction temperature, extraction time, amount of ion-pairing agent and centrifuging time, were investigated and optimized. The calibration curves showed good linear relationship (r>0.9997). The limits of detection and quantification were in the range of 0.052-0.093 and 0.17-0.31 μg mL(-1), respectively. The recoveries were between 70.45% and 94.23% with relative standard deviations lower than 5.31%. The present method is free of volatile organic solvents, and represents lower expenditures of sample, extraction time and solvent, compared with UAE and HRE. There was no obvious difference in the extraction yields of active constitutions obtained by the three extraction methods.

  2. Use of on-site high performance liquid chromatography to evaluate the magnitude and extent of organic contaminants in aquifers

    Science.gov (United States)

    Goerlitz, D.F.; Franks, B.J.

    1989-01-01

    Appraisal of ground water contaminated by organic substances raises problems of difficult sample collection and timely chemical analysis. High-performance liquid chromatography was evaluated for on-site determination of specific organic contaminants in ground water samples and was used at three study sites. Organic solutes were determined directly in water samples, with little or no preparation, and usually in less than an hour after collection. This information improved sampling efficiency and was useful in screening for subsequent laboratory analysis. On two occasions, on-site analysis revealed that samples were undergoing rapid change, with major solutes being upgraded and alteration products being formed. In addition to sample stability, this technique proved valuable for monitoring other sampling factors such as compositional changes with respect to pumping, filtration, and cross contamination. -Authors

  3. High-performance liquid chromatographic method for assay of otilonium bromide, diazepam, and related compounds in finished pharmaceutical forms.

    Science.gov (United States)

    Mannucci, C; Bertini, J; Cocchini, A; Perico, A; Salvagnini, F; Triolo, A

    1993-04-01

    A rapid, simple, stability-indicating assay procedure for otilonium bromide, a smooth muscle relaxant agent, and diazepam in composite tablet analysis was developed with high-performance liquid chromatography. The tablet matrix was dissolved with water, and drugs were extracted with acetonitrile containing an internal standard. An aliquot was centrifuged and chromatographed on a 5-microns, reversed-phase column with 0.5 M sodium acetate trihydrate buffer containing 5 mM 1-heptanesulfonic acid monohydrate sodium salt:methanol (30:70; v/v; adjusted to pH 6.0 with glacial acetic acid) as the mobile phase. The selectivity of the chromatographic system was demonstrated by resolving both compounds from various potential degradation products of each compound. The method is linear, quantitative, and reproducible.

  4. Modernization of Chinese herbal compound and the high performance liquid chromatography tandem mass spectrometry (HPLC-MS)

    Institute of Scientific and Technical Information of China (English)

    LI Wen-lan; SUN Zhi; DU Juan

    2008-01-01

    Chinese herbal compound is playing an important role on curing human diseases. And it has been a trend that Chinese herbal compound is being used all over the world in 21 century. However, our Chinese herbal compound is facing serious challenge for the lack of canonical system of quality criterion for Chinese herbal compound so it has been a urgent problem to set up the quality control standards and reveal therapeutic basis of Chinese herbal compound. In order to give full play to the advantages of Chinese herbal compound, modern scientific and technological is used to research of Chinese herbal compound, especially the high performance liquid chromatography tandem mass spectrometry(HPLC-MS), because it is high sensitive, rapid, and obtain more information. It is very necessary that HPLC-MS is uesed to elucidate the effective components of basic substances of Chinese Herbal Compound, and endow traditional Chinese medicine with modern scientific connotation.

  5. High performance liquid chromatographic method fo pyrantel tartrate in swine feeds and supplements.

    Science.gov (United States)

    Goras, J T

    1981-11-01

    A new method for the determination of pyrantel tartrate in swine feed an supplements has been developed because the current official AOAC method is not applicable to feeds co-medicated with tylosin. The new method involves: (a) leaching of drug from feed with methanolic NaCl solution, (b) removal of interfering substances by ion pair liquid-liquid extraction and high performance liquid chromatography, and (c) quantitation of pyrantel tartrate by monitoring the ultraviolet absorption of the effluent stream at 313nm. The method of standard addition is used to compensate for the effect of the feed matrix on drug recovery. No interference is encountered from tylosin, carbadox, lincomycin, non-drug components of feeds and supplements, or potential degradation products of pyrantel tartrate, i.e., cis isomer of pyrantel tartrate and (E)-N-(3-methylaminopropyl)-2-thiopheneacrylamide. Results for the assay of 3 lots each of feeds and supplements containing 0.0106 and 0.106% pyrantel tartrate, respectively, were within +/-4% of label claim. Coefficients of variation ranged from 1.6 to 1.8% for feeds and from 1.9 to 3.9% for supplements.

  6. Determination of Harmine and Harmaline in Peganum harmala Seeds by High-Performance Liquid Chromatography

    Science.gov (United States)

    Monsef-Esfahani, Hamid Reza; Faramarzi, Mohammad Ali; Mortezaee, Venus; Amini, Mohsen; Rouini, Mohammad Reza

    A High-Performance Liquid Chromatography (HPLC) method was developed for determination of harmine, harmaline, harmol and harmalol in the extract of Peganum harmala seeds. The sample preparation was performed using liquid-liquid extraction. Chromatographic separation was achieved with a Tracer Excel 120 ODSA (150x4.6 mm) column, using a mixture of potassium phosphate buffer (10 mM, pH 7.0): acetonitrile (100:30; v/v) as mobile phase, in an isocratic mode at 1.5 mL min-1. UV detection (λ = 330 nm) was used. The calibration curves were linear (r2>0.998) in the concentration range of 0.5-20 μg mL-1 for all analytes. The lower limit of quantification for all analytes was 0.5 μg mL-1. The within and between day precisions in the measurement of QC samples at three tested concentrations were in the range of 0.6-10.2% for all analytes. The HPLC method is able to measure the harmala alkaloids in the plant extract. The method has suitable reproducibility, sensitivity and resolution for routine and accurate use with UV detection.

  7. Surveillance of the Army’s Propellant Stockpile: Analysis of Stabilizer Content by High Performance Liquid Chromatography

    Science.gov (United States)

    1990-10-01

    high performance liquid chromatography to monitor the level of stabilizer and its daughter products in propellant. Three of those methods are presented along with statistics and some comments about their

  8. HIGH PERFORMANCE LIQUID CHROMATOGRAPHY OF COBALT(III) MIXED LIGAND COMPLEXES WITH ACETYLACETONATE AND OXINATE AND MONOMETHYLOXINATE IONS

    National Research Council Canada - National Science Library

    Kidani, Yoshinori; Naga, Shinobu; Koike, Hisashi

    1976-01-01

    In the presence of Co(III) mixed ligand complexes with acetylacetonate, and oxinate and monomethyloxinate ions, the authors attempted to employ high performance liquid chromatography, HPLC, for the separation and purification...

  9. Sensitive determination of 4-O-methylhonokiol in rabbit plasma by high performance liquid chromatography and application

    Institute of Scientific and Technical Information of China (English)

    Ming-Yue Li; Yu-Hai Tang; Xia Liu; Hai-Yan Lu; Xi-Yan Shi

    2011-01-01

    A novel high performance liquid chromatographic method was developed for the determination of 4-O- methylhonokiol in rabbit plasma and was applied to its pharmacokinetic investigation. Plasma samples were treated by one-fold volume of methanol and acetoni

  10. Pharmacokinetics, tissue distribution, and excretion studies of l-isocorypalmine using ultra high performance liquid chromatography with tandem mass spectrometry.

    Science.gov (United States)

    Wang, Weihui; Liu, Jing; Zhao, Xiaoning; Peng, Yan; Wang, Nannan; Lee, David Y W; Dai, Ronghua

    2017-03-01

    l-Isocorypalmine is a newly identified metabolite of l-tetrahydropalmatine with a unique dual pharmacological profile as a partial dopamine receptor 1 agonist and dopamine receptor 2 antagonist properties for treating cocaine use disorder. The purpose of this study was to explore the pharmacokinetic profiles, tissue distribution, and excretion of l-isocorypalmine in Sprague-Dawley rats. A sensitive and reliable ultra high performance liquid chromatography with tandem mass spectrometry method was developed and validated for determination of l-isocorypalmine in biological samples. The biological samples were extracted by liquid-liquid extraction and separated on a Bonshell ASB C18 column (2.1 × 100 mm, 2.7 μm, Agela) with gradient mobile phase at the flow rate of 0.2 mL/min. The detection was performed by positive electrospray ionization with multiple reaction monitoring mode. Satisfactory linearity, precision, accuracy, extraction recovery, and acceptable matrix effect were achieved. The quantitative method was successfully applied to the pharmacokinetics, tissue distribution, and excretion study of l-isocorypalmine. The results showed that l-isocorypalmine was rapidly distributed, and eliminated from rat plasma and manifested linear dynamics in a dose range of 7.5-15 mg/kg. In addition, the results would be helpful for further clinical reference of l-isocorypalmine as a potential candidate drug for the treatment of cocaine addiction.

  11. High-performance Liquid Chromatographic Determination of Urinary Trans, Trans-Muconic Acid Excreted by Workers Occupationally Exposed to Benzene

    Institute of Scientific and Technical Information of China (English)

    XIA-MIN HU; SHI-ZHEN SONG; FANG-LI YE; LI-WEN LIU

    2006-01-01

    To investigate the relationship between trans, trans-muconic acid (ttMA) as benzene metabolite of occupational workers and benzene concentration in air. Methods A rapid and sensitive high-performance liquid chromatography was developed to determine the level of urinary ttMA. ttMA was extrated from urinary samples in liquid-liquid phase a ODS (2) (5u) column (Φ4.6 mm× 150 mm) and detected at wavelength 264 nm in a UV detector using vanillic acid as an internal standard. The mobile phase was acetaticacid/tetrahydrofuran/methanol/water (v/v, 1:2:10:87). The method was validated with 56 urine samples collected from occupationally benzene-exposed individuals. Results A correlation coefficient (r = 0.9963 ) was found for ttMA ranging 0.10-10.00 μg/mL. The limit of detection was 0.10 μg/mL. The recovery and reproducibility were generally over 90%. There was a positive correlation between ttMA and benzene level in air. The equation was Y=0.859+0.108C (before work, r=-0.6200) or Y=1.980+0.179C (after work, r=0.7930). Conclusion This method can be used to determine and control the level of urinary ttMA in those who are occupationally exposed to benzene.

  12. Determination of some B Vitamins in Sour Cherry Juice Using Dispersive Liquid-liquid Microextraction Followed by High-performance Liquid Chromatography

    Science.gov (United States)

    Parsaei, Parvin; Bahmaei, Manouchehr; Ghannadi, AliReza

    2014-01-01

    Dispersive liquid-liquid microextraction method (DLLME) combined with high-performance liquid chromatography-ultraviolet detection (HPLC-UV) was used to determine thiamine (B1), nicotinamide (B3) and pyridoxine (B6) in sour cherry juice. This method was rapid, simple and sensitive. Separation was accomplished using a C18 column. The optimum chromatographic conditions were found to be: mobile phase consisted of 8% methanol and 92% aqueous phase (1% (V/V) acetic acid water solution); flow rate, 0.7 mL/min; detection wavelength, 260 nm and pH, 3.3. The extraction efficiency of thiamine, nicotinamide and pyridoxine was influenced by factors such as: additional salt effect, the kind and volume of disperser and extraction solvents. In this research, the limit of detection (LOD) and quantification (LOQ) were 0.9 and 3 ng/mL for thiamine, 1.5 and 5 ng/mL for nicotinamide, 0.9 and 3 ng/mL for pyridoxine. The relative standard deviations (RSDs) were less than 2.87% (n=3). An appropriate linear behavior over the observed concentration range was obtained with the value of R²>0.996 for the target vitamins. This method was successfully applied to the sour cherry juice samples. Sour cherry var. Gise (Prunus cerasus var. Gise), which was used in this research, was a local variety of the sour cherry with large stone, double flowers, double fruits, dark red skin and dark red juice. This variety was identified in high altitude areas of Isfahan province after five years of study, since 2005, by Agricultural and Natural Resources Research Center of Isfahan. PMID:25587335

  13. Extraction and determination of hormones in cosmetics by homogeneous ionic liquid microextraction high-performance liquid chromatography.

    Science.gov (United States)

    Kang, Mingqin; Sun, Shuo; Li, Na; Zhang, Daihui; Chen, Mingyan; Zhang, Hanqi

    2012-08-01

    The homogeneous ionic liquid microextraction was applied to the extraction of hormones from cosmetics and the hormones were determined by high-performance liquid chromatography. 1-Hexyl-3-methylimidazolium tetrafluoroborate was used as extraction solvent. Ammonium hexafluorophosphate as used as ion-pairing agent. Several experimental parameters, including the volume of 1-hexyl-3-methylimidazolium tetrafluoroborate, amount of ammonium hexafluorophosphate and sodium chloride, extraction and centrifuge time, and the pH value, were optimized. The limits of detection and quantification for the analytes ranged from 0.03 to 0.24 ng/mL and from 0.10 to 0.79 ng/mL, respectively. The precision for determining the hormones was lower than 5.2%. The proposed method was successfully developed for the determination of hormones in real cosmetic samples.

  14. Determination of Four Pesticides in Soil by Homogeneous Ionic Liquid-based Microextraction Coupled with High-performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    YAN Rui; SHAO Ming-yuan; LIANG Zhen-fen; ZHANG Han-qi; YU Ai-min

    2013-01-01

    Homogeneous ionic liquid microextraction was developed for the simultaneceus extraction of dimethomorph,mefenacet,isoprothiolane and oxadiazon from soil.1-Butyl-3-methylimidazolium tetrafluoroborate was used as extraction solvent,and ammonium hexafluorophosphate was used as ion-pairing agent.High-performance liquid chromatography(HPLC) was employed for separation and determination of the analytes.The calibration curves show good linear relationship(r>0.9988).The recoveries are between 74.2% and 97.9% with relative standard deviations(RSDs) lower than 5.97%.The present method is free of volatile organic solvents,and expenditures of sample,extraction time and solvent are lower,compared with ultrasonic and Soxhlet extraction.There was no obvious difference in the extraction recoveries of pesticides obtained by the three extraction methods.

  15. Determination of captopril by high-performance liquid chromatography with direct electrogenerated chemiluminescence

    Science.gov (United States)

    Sun, Yonghua; Zhang, Zhujun; Zhang, Xinfeng

    2013-03-01

    Captopril exhibit electrogenerated chemiluminescence (ECL) in NaNO3 solution when constant current is exerted. Based on this observation, a direct ECL method coupled with high-performance liquid chromatography (HPLC) separation is developed for determination of captopril in human serum. Factors affected the ECL emission are investigated. Under the optimal conditions, the ECL intensity has a linear relationship with the concentration of captopril in the range of 4.0 × 10-6-2.0 × 10-3 g mL-1 and the detection limit is 2 × 10-6 g mL-1 (S/N = 3). Compared with the common electrogenerated chemiluminescence experiments, the developed method need no any other fluorescence additives.

  16. Analysis of lipophilic pigments from a phototrophic microbial mat community by high performance liquid chromatography

    Science.gov (United States)

    Palmisano, A. C.; Cronin, S. E.; Des Marais, D. J.

    1988-01-01

    As assay for lipophilic pigments in phototrophic microbial mat communities using reverse phase-high performance liquid chromatography was developed which allows the separation of 15 carotenoids and chloropigments in a single 30 min program. Lipophilic pigments in a laminated mat from a commercial salina near Laguna Guerrero Negro, Baja California Sur, Mexico reflected their source organisms. Myxoxanthophyll, echinenone, canthaxanthin, and zeaxanthin were derived from cyanobacteria; chlorophyll c, and fucoxanthin from diatoms; chlorophyll a from cyanobacteria and diatoms; bacteriochlorophylls a and c, bacteriophaeophytin a, and gamma-carotene from Chloroflexus spp.; and beta-carotene from a variety of phototrophs. Sensitivity of detection was 0.6-6.1 ng for carotenoids and 1.7-12 ng for most chloropigments. This assay represents a significant improvement over previous analyses of lipophilic pigments in microbial mats and promises to have a wider application to other types of phototrophic communities.

  17. Simultaneous Determination of Five Components Including Acetaminophen by Reversed-phase High Performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    ZHANG Li-qing; WU Xiao-hua; LU Ying; WANG Xia

    2004-01-01

    High performance liquid chromatography with a C18 reverse-phase column was used to separatethe five components in cough syrup, including acetaminophen, p-aminophenol, caffeine, chlorphenamine maleateand guaifenesin. The mobile phase consists of 15wi% acetonitrile, 0.004mol/L sodium heptyl sulfonate,0.03 mole/L potassium di- hydrogen phosphate and triethylamine ( volume ratio 13: 40: 44: 3), the pH of which isadjusted to 3.0 by phosphoric acid. The contents of the five components are analyzed on an ultraviolet spectropho-tometer at 254nm, with a flow rate of 0.4mL/min. The results show that the calibration curves are linear in acertain range. The average recovery of five components is between 96.31% and 102.3% .

  18. 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.

  19. High performance liquid chromatography study of complex oxygenated alkane mixtures from organic aerosols

    Science.gov (United States)

    Kalafut-Pettibone, Alicia J.; Klems, Joseph P.; McGivern, W. Sean

    2013-05-01

    The composition of secondary organic aerosol particles is of primary importance both in identifying particle sources and in determining physical parameters, such as cloud condensation nucleus propensity. Further, measurements of composition are valuable in determining the chemistry of formation and aging. In this work, we describe the application of a recently-developed derivatization technique to a complex synthetic organic aerosol derived from the photolysis of 1-iodooctane. The technique utilizes high-performance liquid chromatography (HPLC) coupled to both ultravioletvisible (UV/VIS) spectroscopy and tandem mass spectrometry (MS-MS) to determine the overall distribution of hydroxyl (OH), non-acid carbonyl (C=O), and carboxylic acid (COOH) moieties as well as the specific identities of chromatographically separated products. This composition data will then be used to constrain models of the particle formation mechanisms.

  20. The search for more pH stable stationary phases for high performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Collins, Carol H.; Silva, Cesar R.; Faria, Anizio M.; Collins, Kenneth E.; Jardim, Isabel Cristina S.F. [Universidade Estadual de Campinas (UNICAMP), SP (Brazil). Inst. de Quimica

    2009-07-01

    High performance liquid chromatographic (HPLC) separations are largely carried out using reversed phase conditions with stationary phases based on silica. A serious problem with these stationary phases is the tendency of silica to dissolve in high pH solutions often needed to separate basic compounds. The literature reports many different ways that have been tried to resolve this problem. This paper reports the results obtained in our laboratory with stationary phases prepared using silica supports having a layer of a metal oxide (zirconia or titania) attached on their surfaces, followed by immobilization of a polysiloxane or by organofunctionalization with a trimethoxyalkylsilane. Stability tests, also developed in our laboratory, indicate that the metal oxide layer increases the HPLC column lifetimes by making the stationary phase less susceptible to dissolution. (author)

  1. Analysis of carotenoid and porphyrin pigments of geochemical interest by high-performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Hajibrahim, S.K. (Univ. of Bristol, Eng.); Tibbetts, P.J.C.; Watts, C.D.; Maxwell, J.R.; Eglinton, G.; Colin, H.; Guiochon, G.

    1978-04-01

    High-performance liquid chromatography (HPLC) is shown to be a powerful tool in the analysis of carotenoid and porphyrin pigments. Columns packed with 5-..mu..m irregular silica gel particles by a high density and high constant pressure method allow efficient separation of mixtures of total nonsaponifiable carotenoids from recent sedimentary situations. Good reproducibility of retention times (within 2%) is achieved in the gradient elution mode. However, attention must be paid to reequilibration of the column after each injection by washing with the less polar solvent for a minimum of 15 min (for carotenoids) or of 30 min (for porphyrins). HPLC appears to be useful in ''fingerprinting'' petroporphyrin distributions in crude oil.

  2. Simultaneous determination of tricarboxylic acid cycle metabolites by high-performance liquid chromatography with ultraviolet detection.

    Science.gov (United States)

    Shurubor, Yevgeniya I; Cooper, Arthur J L; Isakova, Elena P; Deryabina, Yulia I; Beal, M Flint; Krasnikov, Boris F

    2016-06-15

    Here we describe a simple high-performance liquid chromatography (HPLC) procedure for the simultaneous detection and quantitation in standard solutions of 13 important metabolites of cellular energy metabolism, including 9 tricarboxylic acid (TCA) cycle components and 4 additional metabolites. The metabolites are detected by their absorbance at 210 nm. The procedure does not require prior derivatization, and an analysis can be carried out at ambient temperature within 15 min. The significance of the current work is that the current HPLC procedure should motivate the development of simplified TCA cycle enzyme assays, isotopomer analysis, and determination of selected TCA metabolite levels in plasma/tissues. Copyright © 2016 Elsevier Inc. All rights reserved.

  3. Development of N-ferrocenyl(benzoyl)amino-acid esters stationary phase for high performance liquid chromatography.

    Science.gov (United States)

    Zhou, Xiaohua; Li, Xiaole; Cao, Aijuan; Lijun, Qiao; Yu, Ajuan; Zhang, Shusheng; Wu, Yangjie

    2015-11-01

    A new stationary phase for high-performance liquid chromatography was prepared by covalently bonding N-ferrocenyl(benzoyl)amino-acid esters (L(1)) onto silica gel using 3-aminopropyltriethoxysilane as coupling reagent. The structure of new material was characterized by infrared spectroscopy, elemental analysis and thermogravimetric analysis. The chromatographic behavior of the phase was illustrated in reversed-phase (RP) and normal-phase (NP) modes using polycyclic aromatic hydrocarbons (PAHs), aromatics positional isomers, amines, 5-nitroimidazoles, organophosphorus pesticides and phenols as probes. Multiple mechanisms including hydrophobic, hydrogen bonding, π-π, dipole-dipole, charge-transfer and acid-base equilibrium interactions are involved. Thus, multi-interaction mechanisms and mixed-mode separation of the new phase can very likely guarantee its excellent chromatographic performance for the analysis of complex samples. The L(1) AminoSil column was successfully employed for the analysis of organophosphorus pesticides in vegetable.

  4. Quantitative determination of gibberellins by high performance liquid chromatography from various gibberellins producing Fusarium strains.

    Science.gov (United States)

    Bhalla, Kirti; Singh, Shashi Bala; Agarwal, Rashmi

    2010-08-01

    High performance liquid chromatographic (HPLC) method was developed for analysis of seven gibberellins, i.e., GA3, GA4, GA7, GA3 methyl ester, GA7 methyl ester 3,13 diacetate, GA7 methyl ester, and fusaric acid, using an isocratic system. Method was used for estimation of gibberellins from different Fusarium strains. Gibberellins were extracted from 28 strains of Fusarium, out of which six strains of Fusarium were isolated from soil of different parts of India and 22 strains were procured from the Indian Type Culture Collection, Indian Agricultural Research Institute, New Delhi. Extracts were analyzed for qualitative and quantitative estimation of gibberellins by thin layer chromatography and HPLC, respectively. On the basis of quantitative analysis of produced gibberellins by HPLC, they were categorized as low, moderate, and high gibberellin producing strain. For the first time, Fusarium solani was also reported as high GA3 producing strain.

  5. DETERMINATION OF PROTEINS FROM BUFFALO MILK USING HIGH PERFORMANCE LIQUID CROMATOGRAPHY RP-HPLC

    Directory of Open Access Journals (Sweden)

    AURELIA PECE

    2013-12-01

    Full Text Available In the hereby paper, we have undertaken a study on buffalo milk proteins, employing high performance liquid chromatography (HPLC. This RP-HPLC technique is commonly employed in the separation and assessment of caseins K, and in the fresh, as well as processed milk. These methods are also successfully applied in the authenticity and origin assessment of certain cheese products and the qualitative analysis of milk in bubalines, ovines, caprines and bovines (Ferreira si Cacote, 2003; Veloso si colab., 2002. In order to identify the main protein types, we found support in literature data (Miranda si colab., 2004, and thus achieved the chromatographic separation on whole milk, lactoserum and casein samples.

  6. Creation of reversed phase high-performance liquid chromatographic technique to assay platelet-activating factor

    Institute of Scientific and Technical Information of China (English)

    杨云梅; 曹红翠; 徐哲荣; 陈晓明

    2004-01-01

    Objective: To establish a new assay for platelet-activating factor (PAF), to compare it with bio-assay; and to discuss its significance in some elderly people diseases such as cerebral infarction and coronary heart disease. Methods: To measure PAF levels in 100 controls, 23 elderly patients with cerebral infarction and 65 cases with coronary heart disease by reversed phase high-performance liquid chromatographic technique (rHPLC). Results: rHPLC is more convenient, sensitive, specific, and less confusing, compared with bio-assay. The level of plasma PAF in patients with cerebral infarction was higher than that in the controls (P<0.01), and in patients with coronary heart disease. Conclusion: Detection of PAF with rHPLC is more reliable and more accurate. The new assay has important significance in PAF research.

  7. Creation of reversed phase high-performance liquid chromatographic technique to assay platelet-activating factor

    Institute of Scientific and Technical Information of China (English)

    杨云梅; 曹红翠; 徐哲荣; 陈晓明

    2004-01-01

    Objective: To establish a new assay for platelet-activating factor (PAF), to compare it with bio-assay; and to discuss its significance in some elderly people diseases such as cerebral infarction and coronary heart disease. Methods: To measure PAF levels in 100 controls, 23 elderly patients with cerebral infarction and 65 cases with coronary heart disease by reversed phase high-performance liquid chromatographic technique (rHPLC). Results:rHPLC is more convenient, sensitive,specific, and less confusing, compared with bio-assay. The level of plasma PAF in patients with cerebral infarction was higher than that in the controls (P<0.01), and in patients with coronary heart disease. Conclusion: Detection of PAF with rHPLC is more reliable and more accurate. The new assay has important significance in PAF research.

  8. [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.

  9. High-performance liquid chromatographic analysis of anthraquinone compounds in the Laurera benguelensis

    Science.gov (United States)

    Manojlović, N.; Marković, Z.; Gritsanapan, W.; Boonpragob, K.

    2009-09-01

    A high-performance liquid chromatographic (HPLC) method has been developed for the characterization of anthraquinone metabolites in extracts of the lichen Laurera benguelensis. With this method four anthraquinone derivatives 1,8-dihydroxy-3-methoxy-6-methylanthraquinone, 1,8-dihydroxy-3-formyl-6-methoxyanthraquinone, 1,8-dihydroxy-3-hydroxymethyl-6-methoxy-anthraquinone and 1,3,8-trihyroxy-6-methylanthraquinone can be analyzed. Components of lichen were detected by characteristic ultraviolet spectra and relative retention times. This is first report of phytochemical analysis of L. benguelensis. Importance of this research is in recognizing some new source (lichen and its extracts) as a natural emplacement of antioxidants because oxidation with free radicals or autooxidation is big problem for preservation of food products.

  10. Postcolumn photolysis of pesticides for fluorometric determination by high-performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Miles, C.J.; Moye, H.A.

    1988-02-01

    A high-performance liquid chromatography postcolumn reaction detector that employs UV photolysis with an optional reaction by using o-phthalaldehyde-2-mercaptoethanol (OPA-MERC) followed by fluorescence detection was found to be useful for several classes of pesticides. In the presence of the OPA-MERC reagent, most carbamates, carbamoyl oximes, carbamothioic acids, and substituted ureas gave a sensitive response while the response of dithiocarbamates, phenylamides, and phenylcarbamates varied. The response of most of the pesticides tested was significantly affected by the solvent used. Method detection limits for aldicarb sulfoxide, aldicarb, propoxur, thiram, and neburon in ground water were 2.5, 2.3, 3.3, 3.8, and 2.0 ..mu..g/L, respectively. In the absence of OPA-MERC reagent, several of the substituted aromatic compounds also gave strong fluorescence after photolysis. This detector is applicable to a broad range of nitrogenous pesticides.

  11. 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.

  12. 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.

  13. A simple high performance liquid chromatography method for determination of rebamipide in rat urine.

    Science.gov (United States)

    Cooper, Dustin L; Harirforoosh, Sam

    2014-01-01

    Rebamipide is a mucoprotective agent commonly used to prevent nonsteriodal anti-inflammatory drug-induced gastrointenstinal side effects [1]. Human plasma and urine analysis of rebamipide utilizing high performance liquid chromatography (HPLC) have been reported [2]. Recently, we reported on the plasma levels of rebamipide in presense or absence of celecoxib or diclofenac in rats [3] using a modified HPLC method of detection developed by Jeoung et al. [4]. To tailor the method towards use in urinary rebamipide extraction and analysis, the following modifications were made:•To compensate for high concentrations of rebamipide found in urine, a new rebamipide stock solution was prepared with a final concentration of 50,000 ng/mL.•Rat urine calibration standards were obtained within the range of 50-1000 ng/mL and 1000-50,000 ng/mL.•Plasma samples were replaced with urine samples.

  14. A validated new method for nevirapine quantitation in human plasma via high-performance liquid chromatography.

    Science.gov (United States)

    Silverthorn, Courtney F; Parsons, Teresa L

    2006-01-01

    A fully validated and clinically relevant assay was developed for the assessment of nevirapine concentrations in neonate blood plasma samples. Solid-phase extraction with an acid-base wash series was used to prepare subject samples for analysis. Samples were separated by high performance liquid chromatography and detected at 280 nm on a C8 reverse-phase column in an isocratic mobile phase. The retention times of nevirapine and its internal standard were 5.0 and 6.9 min, respectively. The method was validated by assessment of accuracy and precision (statistical values 0.996) and the average recovery was 93% (n = 18). The lower limit of quantification (relative standard deviation <20%) was determined to be 25 ng/mL for 50 microL of plasma, allowing detection of as little as 1.25 ng of nevirapine in a sample. This value represents an increase in sensitivity of up to 30-fold over previously published methods.

  15. Determination of Sex Hormones in Antler Velvet by High Performance Liquid Chromatography Tandem Mass Spectrometry

    Institute of Scientific and Technical Information of China (English)

    LU Chun-mei; WANG Ming-tai; MU Jun; BAI Yu-ping; DU Jian-shi; ZHANG Han-qi; WANG Jian-wei

    2012-01-01

    Eighteen sex hormones in antler velvet were determined by high performance liquid chromatography tandem mass spectrometry.The solid phase extraction was applied to eliminating the matrix effect.The experimental conditions were examined and optimized.Under the optimal conditions,the proposed method provides the good linearities and determination limits(0.2-1.0 μg/kg)of the analytes investigated.The recoveries ranging from 72.3% to 149.5% were obtained for the target analytes at two concentration levels.This method was applied to the determination of eighteen sex hormones in different kinds of antler velvet samples and the obtained results are satisfactory.The results indicate that the proposed method is suitable for the determination of sex hormones in antler velvet samples.

  16. Improved high-performance liquid chromatographic analysis of terazosin in human plasma.

    Science.gov (United States)

    Cheah, P Y; Yuen, K H; Liong, M L

    2000-08-18

    A simple, sensitive and reproducible high-performance liquid chromatography (HPLC) method was developed for the determination of terazosin in human plasma. The method involves a one-step single solvent extraction procedure using dichloromethane with a 0.25 ml plasma sample. Recovery values were all greater than 90% over the concentration range 0.25-100 ng/ml. Terazosin was found to adsorb to glass or plastic tubes, but this could be circumvented by using disposable plastic tubes. Also, rinsing the injector port with methanol after each injection helped to prevent any carry-over effect. The internal standard, prazosin, did not exhibit this problem. The method has a quantification limit of 0.25 ng/ml. The within- and between-day coefficient of variation and accuracy values were all less than 7% over the concentration range 0.25-100 ng/ml and hence the method is suitable for use in pharmacokinetic studies of terazosin.

  17. Measurement of serum pralidoxime methylsulfate (Contrathion) by high-performance liquid chromatography with electrochemical detection.

    Science.gov (United States)

    Houzé, Pascal; Borron, Stephen W; Scherninski, François; Bousquet, Bernard; Gourmel, Bernard; Baud, Frédéric

    2005-01-05

    Pralidoxime methylsulfate (Contrathion) is widely used to treat organophosphate poisoning. Despite animal and human studies, the usefulness of Contrathion therapy remains a matter of debate. Therapeutic dosage regimens need to be clarified and availability of a reliable method for plasma pralidoxime quantification would be helpful in this process. We here describe a high-performance liquid chromatography technique with electrochemical detection to measure pralidoxime concentrations in human serum using guanosine as an internal standard. The assay was linear between 0.25 and 50 microg mL(-1) with a quantification limit of 0.2 microg mL(-1). The analytical precision was satisfactory, with variation coefficients lower 10%. This assay was applied to the analysis of a serum from an organophosphorate poisoned patient and treated by Contrathion infusions (100 and 200 mg h(-1)) after a loading dose (400 mg).

  18. Determination of triazine herbicides in milk by cloud point extraction and high-performance liquid chromatography.

    Science.gov (United States)

    Liu, Tiantian; Cao, Peng; Geng, Jinpei; Li, Jinqiang; Wang, Mingzhen; Wang, Minglin; Li, Xiaoyu; Yin, Dalu

    2014-01-01

    High-performance liquid chromatography with UV detection was used to detect four triazines in milk. An efficient pretreatment method known as cloud point extraction (CPE) was proposed for extracting and preconcentrating analytes. The parameters of CPE including surfactant type and concentration, electrolyte, sample pH, incubation temperature and duration were investigated. Under optimal conditions, satisfying recoveries in the range of 70.5-96.9% were achieved for four triazines. The limits of detection ranged from 6.79 to 11.19μg L(-1). The linear range of quantitation for the four triazines was 50-2000μg L(-1), and the correlation coefficients of the calibration curves were all 0.9999. The results demonstrated that the proposed method was efficient and reliable for the determination of triazine herbicides in milk samples.

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

    Institute of Scientific and Technical Information of China (English)

    Zheng-Xiang Zhang; Xue Qiao; Min Ye; Man-Yu Zhang; Yue Song; Tao Bo

    2015-01-01

    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.

  20. High-performance liquid chromatographic determination of alpha-tocopherol in macroalgae.

    Science.gov (United States)

    Sánchez-Machado, D I; López-Hernández, J; Paseiro-Losada, P

    2002-11-08

    A high-performance liquid chromatographic (HPLC) method for the microscale determination of alpha-tocopherol in macroalgae is reported. The method includes microscale saponification and extraction with n-hexane. The presence of alpha-tocopherol in macroalgae samples was confirmed by HPLC-MS. Alpha-tocopherol levels as determined in samples by HPLC with UV and fluorescence detection did not differ significantly; however, fluorescence detection has a higher sensitivity (detection limit 10.4 ng/ml, vs. 104 ng/ml with UV detection), as well as good precision (relative standard deviation 1.81%) and recovery (94.3%). Fluorescence detection is also faster. We used this method to determine the alpha-tocopherol contents of four commercial macroalgae products from northwest Spain as part of nutritional studies in dehydrated Himanthalia elongata and Laminaria ochroleuca, and also in canned Himanthalia elongata and Saccorhiza polychides.

  1. High-Performance Liquid Chromatographic Method for Determination of Germacrone in Rat Plasma

    Institute of Scientific and Technical Information of China (English)

    HEHai-bing; TANGXing; CUIFu-de

    2004-01-01

    Aim A reliable high-performance liquid chromatographic (HPLC) method was developed for determination of gennacrone in rat. plasma. Methods The plasma samples were treated with acetonitrile and analyzed by HPLC with UV detection at 244 nm. Results The limit of detection was 100 ng·mL-1 for germacrone in plasma and the linear range was0.1004-15.06μg· mL-1 in plasma. The RSD of intra-day and inter-day assay was 1.87%-4.29% and 1.29%-5.15%, respectively. The recoveries of germacrone were over 95%. The endogenous substances in plasma did not show any interference in the analysis. Conclusion The method is accurate and convenient, and suitable for pharmacokinetic studies of gennacrone in rats.

  2. Detection of cheese whey and caseinomacropeptide in fermented milk beverages using high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    E.H.P. Andrade

    2014-06-01

    Full Text Available Cheese whey level and caseinomacropeptide (CMP index of fermented milk beverages added with four levels of cheese whey (0, 10, 20, and 40% and stored at 8-10oC for 0, 7, 14 and 21 days were determined by high performance liquid chromatography-gel filtration (HPLC-GF. Additionally, the interference of the starter culture and the storage time on the detection of cheese whey and CMP were investigated. Refrigerated storage up to 21 days did not affect (P>0.05 cheese whey and CMP amounts in milk (0% of cheese whey and in fermented milk beverages added with 10 and 20% of cheese whey (P>0.05. However, cheese whey and CMP amounts were higher than expected (P<0.05 in fermented milk beverages added with 40% of cheese whey and stored for 21 days.

  3. Tritium isotope effect in high-performance liquid chromatography of eicosanoids

    Energy Technology Data Exchange (ETDEWEB)

    Do, U.H.; Lo, S.-L.; Iles, J.; Rosenberger, T.; Tam, P.; Hong, Y.; Ahern, D. [DuPont, NEN Products, Boston (United States). Lipids Group

    1994-07-01

    A significant difference in retention time between unlabeled and the corresponding multi-tritium-labeled eicosanoid has been observed in the high performance liquid chromatography (HPLC) analysis of 11 eicosanoids. Variations in retention time range from 3-7%, depending on the separation conditions as well as the number and position of the tritium substitution. Multi-tritium-labeled eicosanoids were eluted earlier than the corresponding unlabeled eicosanoid in reversed phase HPLC, whereas no isotope effect was seen with {sup 14}C- and {sup 3}H{sub 2}-eicosanoids. Considerations must be given to this tritium isotope effect whenever both multi-tritium-labeled and unlabeled eicosanoids are used for HPLC cochromatography or recovery studies. (author).

  4. 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...... 150 microliters whole blood dried on filter paper was found to be 20 ng/ml for DDS and PYR and 15 ng/ml for MADDS (precision ... paper method and the original whole-blood method was almost comparable. Standardization could therefore be obtained by the more simple whole-blood method. Dried filter paper samples stored at 19-22 degrees C were stable for months and for 2 weeks stored at 35 degrees C. The concentrations...

  5. Determination of primary amino acids in wines by high performance liquid magneto-chromatography.

    Science.gov (United States)

    Barrado, E; Rodriguez, J A; Castrillejo, Y

    2009-05-15

    Eight amino acids (ethanolamine, glycine, alanine, beta-aminobutyric acid, leucine, methionine, histidine and asparagine) were identified and quantified in Spanish wines by high performance liquid magneto-chromatography (HPLMC) with UV-V spectrophotometry. For this method, the amino acids are first complexed with mono(1,10-phenanthroline)-Cu(II) to confer them paramagnetic properties, and then separated by application of a low magnetic field intensity (5.5 mT) to the stationary phase contained in the chromatographic column. Principal components analysis of the results obtained grouped together the wine samples according to their denomination of origin: "Ribera del Duero", "Rueda" or "Rioja" (Spain). Through cluster analysis, a series of correlations was also observed among certain amino acids, and between these groupings and the type of wine. These clusters were found to reflect the role played by the amino acids as primary or secondary nutrients for the bacteria involved in alcoholic and malolactic fermentation.

  6. Rocket-powered high-performance liquid chromatographic analysis of plant ascorbate and glutathione.

    Science.gov (United States)

    Davey, M W; Dekempeneer, E; Keulemans, J

    2003-05-01

    We describe a robust procedure for the extraction and high-performance liquid chromatographic analysis of L-ascorbate (vitamin C), glutathione (gamma-glutamyl cysteinylglycine), and their respective oxidized forms from various plant tissues. Parameters such as the choice of extraction buffer, tissue disruption technique, sample stability, and separation conditions have all been optimized. In particular we found that the inclusion of the reducing agent dithiothreitol as a "stabilizer" in extracts with high phenolic content actually promoted oxidation of these antioxidants. Further, by using commercially available short "Rocket" HPLC columns in combination with high mobile-phase flow rates, analysis times were reduced to only 6min, making the method suitable for the high-resolution screening of large numbers of samples.

  7. 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.

  8. 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.

  9. High-performance liquid chromatographic determination of vertilmicin in rat plasma using sensitive fluorometric derivatization.

    Science.gov (United States)

    Liu, Zhen; Sha, Yunfei; Huang, Taomin; Yang, Bei; Duan, Geng-Li

    2005-12-15

    A sensitive and reliable high-performance liquid chromatographic method was developed for the determination of vertilmicin in rat plasma. Derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) followed by C(18) reversed-phase chromatography allowed the fluorimetric detection of vertilmicin. Optimal conditions for the derivatization of vertilmicin are described. The limit of quantification was 0.02 mg/L. The pharmacokinetics of vertilmicin was studied in 24 rats following intramuscular injection (i.m.) of different doses (4, 8, 16, 32 mg/kg of body weight). The pharmacokinetic parameter values were estimated by use of 3P97 program. In this study, we assessed the dose proportionality of vertilmicin after single intramuscular injection doses and obtained new information on the pharmacokinetics of the compound.

  10. Determination of artificial sweeteners in beverages and special nutritional products using high performance liquid chromatography.

    Science.gov (United States)

    Serdar, Maja; Knežević, Zorka

    2011-06-01

    This paper presents two high performance liquid chromatographic (HPLC) methods used for the separation and determination of artificial sweeteners aspartame, acesulphame K, sodium saccharin, and sodium cyclamate in beverages and special nutritional products (special food intended for specific population groups). All four compounds are soluble in aqueous solutions and can easily be separated and determined by HPLC with a diode array detector (DAD). The first method involved separation of aspartame, acesulphame K, and sodium saccharin on a C18 column with an isocratic elution of phosphate buffer and acetonitrile as mobile phase. The second method was used to separate sodium cyclamate on a C18 column with methanol and water as mobile phase. Under optimum conditions, both methods showed good analytical performance, such as linearity, precision, and recovery. The methods were successfully applied for the analysis of real samples of soft drinks and special nutritional products.

  11. 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.

  12. High Performance Liquid Chromatography of the Dansyl Derivatives of Putrescine, Spermidine, and Spermine 1

    Science.gov (United States)

    Escribano, M. Isabel; Legaz, M. Estrella

    1988-01-01

    A high performance liquid chromatographic (HPLC) method, based on dansylation and fluorescence detection, is described for the estimation of putrescine, spermidine, and spermine in lichen (Evernia prunastri [L.]) samples. Because of the high concentrations of phenols and salts, dansylation was followed by a pre-HPLC purification step. Both flow rate and mobile phase (methanol:water) followed a gradient for optimum resolution on a reverse-phase column. Amounts as small as 0.3 picomole of standard polyamines could be detected. In applying the method to lichens, it was found that 5.45% (w/w) of the exogenous putrescine taken up by the thallus was unbound in the algal partner and that 60% (w/w) was conjugated in the thallus, perhaps to lichen phenolics. PMID:16666175

  13. High performance liquid chromatography of the dansyl derivatives of putrescine, spermidine, and spermine.

    Science.gov (United States)

    Escribano, M I; Legaz, M E

    1988-06-01

    A high performance liquid chromatographic (HPLC) method, based on dansylation and fluorescence detection, is described for the estimation of putrescine, spermidine, and spermine in lichen (Evernia prunastri [L.]) samples. Because of the high concentrations of phenols and salts, dansylation was followed by a pre-HPLC purification step. Both flow rate and mobile phase (methanol:water) followed a gradient for optimum resolution on a reverse-phase column. Amounts as small as 0.3 picomole of standard polyamines could be detected. In applying the method to lichens, it was found that 5.45% (w/w) of the exogenous putrescine taken up by the thallus was unbound in the algal partner and that 60% (w/w) was conjugated in the thallus, perhaps to lichen phenolics.

  14. High-performance liquid chromatographic assay detects pentamidine metabolism by Fisher rat liver microsomes.

    Science.gov (United States)

    Tuttle, R H; Hall, J E; Tidwell, R R

    1997-01-24

    Fisher rat liver microsomes metabolized the antimicrobial drug pentamidine to four new compounds detected by gradient elution reversed-phase high-performance liquid chromatography with variable wavelength detection. Coelution experiments with pentamidine metabolite standards determined the new peaks to be previously identified hydroxylated metabolites of pentamidine, with 1,5-bis(4'-amidinophenoxy)-3-pentanol and 1,5-di-(4'-amidinophenoxy)-2-pentanol formed in the greatest amount. The data contradict a previous report that Fisher rat liver homogenates do not metabolize pentamidine. Pentamidine and its known primary metabolites have almost identical absorption spectra; thus, pentamidine metabolism must be evaluated using gradient elution HPLC to resolve pentamidine from its metabolites. The current assay has now been used to demonstrate that Fisher and Sprague-Dawley rat, mouse, rabbit and human liver microsomes all metabolize pentamidine in vitro.

  15. Microwave-assisted ionic liquid homogeneous liquid-liquid microextraction coupled with high performance liquid chromatography for the determination of anthraquinones in Rheum palmatum L.

    Science.gov (United States)

    Wang, Zhibing; Hu, Jianxue; Du, Hongxia; He, Shuang; Li, Qing; Zhang, Hanqi

    2016-06-01

    The microwave-assisted ionic liquid homogeneous liquid-liquid microextraction (MA-IL-HLLME) coupled with high performance liquid chromatography with diode array detection (HPLC-DAD) was developed for the determination of anthraquinones, including aloe-emodin, emodin, chrysophanol and physcion in root of Rheum palmatum L. Several experimental parameters influencing the extraction efficiency, including amount of sample, type and volume of ionic liquid, volume and pH value of extraction medium, microwave power and extraction time, concentration of NH4PF6 as well as centrifugal condition were optimized. When 140μL of ionic liquid ([C8MIM][BF4]) was used as an extraction solvent, target analytes can be extracted from sample matrix in one minute with the help of microwave irradiation. The MA-IL-HLLME is simple and quick. The calibration curves exhibited good linear relationship (r>0.9984). The limits of detection and quantification were in the range of 0.015-0.026 and 0.051-0.088μgmL(-1), respectively. The spiked recovery for each analyte was in the range of 81.13-93.07% with relative standard deviations lower than 6.89%. The present method is free of volatile organic solvents, and represents lower expenditures of sample, extraction time and solvent, compared with ultrasonic and heat reflux extraction. The results indicated that the present method can be successfully applied to the determination of anthraquinones in medicinal plant.

  16. [Determination of five triazine herbicides in infant milk powder by high performance liquid chromatography coupled with ionic liquid-based homogeneous liquid-liquid microextraction].

    Science.gov (United States)

    Zhang, Liyuan; Yao, Di; Li, Na; Zhang, Hanqi; Yu, Aimin

    2015-07-01

    A high performance liquid chromatography coupled with homogeneous liquid-liquid microextraction was developed for the determination of five triazine herbicides in infant milk powders. The ionic liquid was used as microextraction solvent. The separation of the herbicides was performed on an Eclipse XDB-C18 column using acetonitrile and water as mobile phases in gradient mode. The effects of homogeneous liquid-liquid extraction conditions on the experimental results were investigated in detail. Under the optimized experimental conditions, the calibration curves for determining the analytes were linear and the correlation coefficients were ≥ 0.9992. The limits of detection for cyanazine, desmetryn, terbumeton, terbuthylazine and dimethametryn were 12.1, 13.8, 11.8, 14.6 and 13.7 μg/kg, respectively. The recoveries of the analytes spiked in four infant milk powders ranged from 92.2% to 103.2% and the relative standard deviations were lower than 6%. This method is sensitive, simple, and suitable for the determination of triazine herbicides in milk powder samples.

  17. Compared the New Molecular Method for Rapid Detection of Avian Leukemia Virus by Using Denaturing High Performance Liquid Chromatography Combined with Nucleic Acid Amplification and Real-time PCR%PCR结合变性高效液相色谱法与荧光定量PCR法在检测禽白血病中的比较与应用

    Institute of Scientific and Technical Information of China (English)

    孙涛; 张太翔; 徐彪; 梁成珠; 朱来华; 岳志芹

    2011-01-01

    Compared the new molecular method for rapid detection of avian leukemia virus by using denaturing high performance liquid chromatography(DHPLC) combined with nucleic acid amplification and Real-time PCR in this study. According to the sequence of pol gene of ALV, one pair of primers and the TaqMan probe were designed by using Primer Premier 5. 0. The PCR fragment which was amplified by the primers were analysised by DHPLC and the results of Real-time PCR by the primers and the TaqMan probe. They all compared to normal chicken embryo allantoic fluid, duck plague virus(DPV),infectious bronchitis virus(IBV) ,goose parvovirus(GPV) ,avian influenza virus(H5Nl AIV),Newcastle disease virus(NDV), infectious bursal disease virus (IBDV) ,EDSV. There were tested to confirm the specificity of the PCR-DHPLC assay and no positive absorption peaks occurred. The detection limit of ALV AV228 by PCR-DHPLC was 3 pg,10 fold iower than the ordinary Realtime PCR. The results of detcting organ samples from the chickens were tested by PCR-DHPLC and Real-time PCR,showing 100% agreement.%本研究旨在比较PCR结合变性高效液相色谱技术(PCR-DHPLC)与荧光定量PCR (Real-time PCR)两种方法在检测禽白血病中的应用.根据禽白血病pol基因序列,设计1对引物和1条探针,利用引物进行禽白血病模板的RT-PCR扩增,产物经变性高效液相色谱上样处理;利用引物及探针进行荧光定量PCR扩增,结果与PCR-DHPLC进行比对.两种方法同时用正常鸡胚尿囊液、鸭瘟病毒、传染性支气管炎病毒、鹅细小病毒、H5N1亚型禽流感病毒、新城疫病毒、传染性法氏囊病毒、减蛋综合症病毒做特异性检测;以稀释成不同梯度的AV228毒株核酸做敏感性检测.试验结果表明PCR-DHPLC方法只对禽白血病病原有阳性扩增的吸收峰,Real-time PCR也只对禽白血病病原有阳性扩增,两法均对其他禽源病毒核酸无特异性扩增;PCR-DHPLC与Real-time PCR法

  18. Flavonoid profiling of a traditional Chinese medicine formula of Huangqin Tang using high performance liquid chromatography

    Science.gov (United States)

    Li, Tao; Zhuang, Shuaixing; Wang, Yiwei; Wang, Yanli; Wang, Weihao; Zhang, Huihui; Chen, Li; Wang, Dunfang; Zhou, Zhongming; Yang, Weipeng

    2016-01-01

    The quality control processes for herbal medicines have been problematic. Flavonoids are the major active components of Huangqin Tang (HQT, a traditional Chinese medicine formula). In this study, we used a combinative method approach consisting of chromatographic fingerprinting (high performance liquid chromatography; HPLC), quantitative methods and a pharmacodynamic evaluation model to analyze the flavonoids of HQT obtained from different sources. Ten batches of HQT were analyzed by the HPLC fingerprinting method and 26 common peaks were detected, of which 23 peaks corresponded with the chemical profile of HQT. In addition, 11 major compounds were identified by LC–MS analysis (liquid chromatography–tandem mass spectrometer; LC–MSn) and quantified by the HPLC quantitative method approach. The studied 10 batches of HQT were found to be homogeneous in their composition with a similarity between 0.990 and 1.000. The distribution of the 11 identified compounds was found to be very similar among the batches. Only slight pharmacodynamic differences were detected between the different batches, confirming the homogeneity of HQT. The results of this study prove that the combination of chromatographic fingerprinting and quantitative analysis can be readily used for comprehensive quality control of herbal medicines. PMID:27006899

  19. Fully automated high-performance liquid chromatographic assay for the analysis of free catecholamines in urine.

    Science.gov (United States)

    Said, R; Robinet, D; Barbier, C; Sartre, J; Huguet, C

    1990-08-24

    A totally automated and reliable high-performance liquid chromatographic method is described for the routine determination of free catecholamines (norepinephrine, epinephrine and dopamine) in urine. The catecholamines were isolated from urine samples using small alumina columns. A standard automated method for pH adjustment of urine before the extraction step has been developed. The extraction was performed on an ASPEC (Automatic Sample Preparation with Extraction Columns, Gilson). The eluate was collected in a separate tube and then automatically injected into the chromatographic column. The catecholamines were separated by reversed-phase ion-pair liquid chromatography and quantified by fluorescence detection. No manual intervention was required during the extraction and separation procedure. One sample may be run every 15 min, ca. 96 samples in 24 h. Analytical recoveries for all three catecholamines are 63-87%, and the detection limits are 0.01, 0.01, and 0.03 microM for norepinephrine, epinephrine and dopamine, respectively, which is highly satisfactory for urine. Day-to-day coefficients of variation were less than 10%.

  20. DETERMINATION OF GAMMA - AMINOBUTYRIC ACID BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

    Directory of Open Access Journals (Sweden)

    D. Qujeq

    1998-08-01

    Full Text Available Gamma-aminobutyric acid is the main inhibitory amino acid neurotransmitter in several areas of the brain. Measurement of gamma-aminobutyric acid in the brain will help elucidate its metabolic rote and diagnostic value. The liquid chromatography (LC system consisted of a delivery pump. Male rats (150-250g were used in this study. The brain was rapidly removed from decapitated animals. Trie chromatograms showed satisfactory resolution and symmetrical peak shapes, both with the standard and tissue extracts. the retention lime was 4.60 min. it was constant in repeated analyses and exactly the same for the standard and tissue samples.

  1. [Measurement of 11 benzophenone ultraviolet-filters in cosmetics by high performance liquid chromatography].

    Science.gov (United States)

    Qu, Baocheng; Bian, Haitao; Mao, Xiqin; Li, Jin

    2015-12-01

    A sample preparation and analytical method with liquid-liquid extraction (LLE) and high performance liquid chromatography (HPLC) was developed to detect 11 benzophenone ultraviolet-filters in cosmetics. The target compounds were extracted by the mixed solutions of tetrahydrofuran (TH)/methanol/water or dichloromethane/water at proper ratios. The extracts were centrifuged and filtered to remove matrix compounds, and then analyzed by HPLC. The separation of analytes was carried out on a Diamonsil-C18 column (150 mm x 4.6 mm, 5 μm) with 0.1% (v/v) formic acid aqueous solution (containing 10 mmol/L ammonium acetate) as mobile phase A and methanol containing 0.1% (v/v) formic acid as mobile phase B. The spiked recoveries of the method (n = 7) were 93.4%-103.8% with the relative standard deviations of 0.1%-4.2%. The limits of detection (LODs) were in the range of 4.0-30 μg/g and the limits of quantitation (LOQs) ranged from 15 to 100 μg/g. The method was applied to the determination of 42 cosmetic samples randomly purchased from the supermarket in Dalian. Five benzophenone series were always detected, in which the content of benzophenone-3 in sunscreen cream and the content of benzophenone-2 in perfume were very high and reached 2 785 μg/g and 2 106 μg/g, respectively. The results showed that the developed method is efficient, reliable and sensitive, which can be applied to the determination of benzophenones in cosmetics.

  2. Anatomical study of secondary tuberized roots of Harpagophytum procumbens DC and quantification of harpagoside by high-performance liquid chromatography method

    OpenAIRE

    Fatiha El Babili; Fouraste, I; C Rougaignon; Moulis, C.; Chatelain, C.

    2012-01-01

    Aim and Background: A botanical study is conducted to provide a standard diagnostic tool. In order to improve the quality assurance of the secondary tuberized roots of Harpagophytum procumbens, derived extract and phytomedicine, a simple, rapid, and accurate high-performance liquid chromatography (HPLC) method was developed to assess the harpagoside. Material and Mehods: This HPLC assay was performed on a reversedphase C18 column with methanol and water (50/50–V/V) as the mobile phase with a ...

  3. Analytical Method Validation of High-Performance Liquid Chromatography and Stability-Indicating Study of Medroxyprogesterone Acetate Intravaginal Sponges.

    Science.gov (United States)

    Batrawi, Nidal; Wahdan, Shorouq; Abualhasan, Murad

    2017-01-01

    Medroxyprogesterone acetate is widely used in veterinary medicine as intravaginal dosage for the synchronization of breeding cycle in ewes and goats. The main goal of this study was to develop reverse-phase high-performance liquid chromatography method for the quantification of medroxyprogesterone acetate in veterinary vaginal sponges. A single high-performance liquid chromatography/UV isocratic run was used for the analytical assay of the active ingredient medroxyprogesterone. The chromatographic system consisted of a reverse-phase C18 column as the stationary phase and a mixture of 60% acetonitrile and 40% potassium dihydrogen phosphate buffer as the mobile phase; the pH was adjusted to 5.6. The method was validated according to the International Council for Harmonisation (ICH) guidelines. Forced degradation studies were also performed to evaluate the stability-indicating properties and specificity of the method. Medroxyprogesterone was eluted at 5.9 minutes. The linearity of the method was confirmed in the range of 0.0576 to 0.1134 mg/mL (R(2) > 0.999). The limit of quantification was shown to be 3.9 µg/mL. Precision and accuracy ranges were found to be %RSD <0.2 and 98% to 102%, respectively. Medroxyprogesterone capacity factor value of 2.1, tailing factor value of 1.03, and resolution value of 3.9 were obtained in accordance with ICH guidelines. Based on the obtained results, a rapid, precise, accurate, sensitive, and cost-effective analysis procedure was proposed for quantitative determination of medroxyprogesterone in vaginal sponges. This analytical method is the only available method to analyse medroxyprogesterone in veterinary intravaginal dosage form.

  4. Analytical Method Validation of High-Performance Liquid Chromatography and Stability-Indicating Study of Medroxyprogesterone Acetate Intravaginal Sponges

    Directory of Open Access Journals (Sweden)

    Nidal Batrawi

    2017-02-01

    Full Text Available Medroxyprogesterone acetate is widely used in veterinary medicine as intravaginal dosage for the synchronization of breeding cycle in ewes and goats. The main goal of this study was to develop reverse-phase high-performance liquid chromatography method for the quantification of medroxyprogesterone acetate in veterinary vaginal sponges. A single high-performance liquid chromatography/UV isocratic run was used for the analytical assay of the active ingredient medroxyprogesterone. The chromatographic system consisted of a reverse-phase C18 column as the stationary phase and a mixture of 60% acetonitrile and 40% potassium dihydrogen phosphate buffer as the mobile phase; the pH was adjusted to 5.6. The method was validated according to the International Council for Harmonisation (ICH guidelines. Forced degradation studies were also performed to evaluate the stability-indicating properties and specificity of the method. Medroxyprogesterone was eluted at 5.9 minutes. The linearity of the method was confirmed in the range of 0.0576 to 0.1134 mg/mL ( R 2 > 0.999. The limit of quantification was shown to be 3.9 µg/mL. Precision and accuracy ranges were found to be %RSD <0.2 and 98% to 102%, respectively. Medroxyprogesterone capacity factor value of 2.1, tailing factor value of 1.03, and resolution value of 3.9 were obtained in accordance with ICH guidelines. Based on the obtained results, a rapid, precise, accurate, sensitive, and cost-effective analysis procedure was proposed for quantitative determination of medroxyprogesterone in vaginal sponges. This analytical method is the only available method to analyse medroxyprogesterone in veterinary intravaginal dosage form.

  5. Combining Laser Ablation/Liquid Phase Collection Surface Sampling and High-Performance Liquid Chromatography Electrospray Ionization Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Ovchinnikova, Olga S [ORNL; Kertesz, Vilmos [ORNL; Van Berkel, Gary J [ORNL

    2011-01-01

    This paper describes the coupling of ambient pressure transmission geometry laser ablation with a liquid phase sample collection method for surface sampling and ionization with subsequent mass spectral analysis. A commercially available autosampler was adapted to produce a liquid droplet at the end of the syringe injection needle while in close proximity to the surface to collect the sample plume produced by laser ablation. The sample collection was followed by either flow injection or a high performance liquid chromatography (HPLC) separation of the extracted components and detection with electrospray ionization mass spectrometry (ESI-MS). To illustrate the analytical utility of this coupling, thin films of a commercial ink sample containing rhodamine 6G and of mixed isobaric rhodamine B and 6G dyes on glass microscope slides were analyzed. The flow injection and HPLC/ESI-MS analysis revealed successful laser ablation, capture and, with HPLC, the separation of the two compounds. The ablated circular area was about 70 m in diameter for these experiments. The spatial sampling resolution afforded by the laser ablation, as well as the ability to use sample processing methods like HPLC between the sample collection and ionization steps, makes this combined surface sampling/ionization technique a highly versatile analytical tool.

  6. Quantitative Analysis of Sphingomyelin by High-Performance Liquid Chromatography after Enzymatic Hydrolysis

    Directory of Open Access Journals (Sweden)

    Seunghyun Lee

    2012-01-01

    Full Text Available Sphingomyelin is the most abundant sphingolipid in mammalian cells and is mostly present in the plasma membrane. A new analytical method using high-performance liquid chromatography (HPLC was developed to quantify sphingomyelin in mouse plasma and tissues, 3T3-L1 cells, rat aortic smooth muscle cells, and HT-29 cells. Sphingomyelin and dihydrosphingomyelin, an internal standard, were separated by high-performance thin-layer chromatography and simultaneously hydrolyzed with sphingolipid ceramide N-deacylase and sphingomyelinase to release sphingosine and dihydrosphingosine, respectively. Sphingomyelin content was measured by HPLC following o-phthalaldehyde derivatization. Sphingomyelin concentrations in 3T3-L1 cells, rat aortic smooth muscle cells, and HT-29 cells were 60.10±0.24, 62.69±0.08, and 58.38±0.37 pmol/μg protein, respectively, whereas those in brain, kidney, and liver of ICR mice were 55.60±0.43, 43.75±0.21, and 22.26±0.14 pmol/μg protein. The sphingomyelin concentration in mouse plasma was 407.40±0.31 μM. The limits of detection and quantification for sphingomyelin were 5 and 20 pmol, respectively, in the HPLC analysis with fluorescence detection. This sensitivity was sufficient for analyzing sphingomyelin in biological samples. In conclusion, this analytical method is a sensitive and specific technique for quantifying sphingomyelin and was successfully applied to diverse biological samples with excellent reproducibility.

  7. 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.

  8. Ionic Liquid-based Microwave-assisted Liquid-liquid Microextraction and High Performance Liquid Chromatography Determination of Sulfonamides from Animal Oils

    Institute of Scientific and Technical Information of China (English)

    FENG Xu-dong; LIANG Fang-hui; SU Rui; WU Li-jie; LI Xue-yuan; WANG Xing-hua; ZHANG Han-qi

    2013-01-01

    The authors performed ionic liquid-based microwave-assisted liquid-liquid microextraction(IL-based MALLME) coupled with high performance liquid chromatographic separation for the determination of 6 sulfonamides (SAs) from animal oils.The target analytes were extracted from animal oil samples with sodium hydroxide solution containing 1-butyl-3-methylimidazolium tetrafluoroborateand as the extraction solvent under microwave irradiation.The experimental parameters of the IL-based MALLME,including types of ILs,volume of IL,amount of ion-pairing agent(NH4PF6),pH value of sample solution,and extraction temperature and time were evaluated.The limits of detection and quantification obtained were in a range of 0.4-0.5 μg/kg and a range of 1.2-1.8 μg/kg,respectively.The accuracy of the method was evaluated by analyzing five spiked animal oil samples at two fortified levels(5 and 50 μg/kg),and the recoveries of SAs varied from 81.4% to 114.5% with relative standard deviations ranging from 0.8% to 9.0%.

  9. Quantitation of antioxidants in water samples using ionic liquid dispersive liquid-liquid microextraction followed by high-performance liquid chromatography-ultraviolet detection.

    Science.gov (United States)

    Sobhi, Hamid Reza; Kashtiaray, Amir; Farahani, Hadi; Farahani, Mohammad Reza

    2011-01-01

    A simple and efficient method, ionic liquid-based dispersive liquid-liquid microextraction combined with high-performance liquid chromatography-ultraviolet detection (HPLC-UV), has been applied for the extraction and determination of some antioxidants (Irganox 1010, Irganox 1076 and Irgafos 168) in water samples. The microextraction efficiency factors were investigated and optimized: 1-hexyl-3-methylimidazolium hexafluorophosphate [C(6)MIM][PF(6)] (0.06 g) as extracting solvent, methanol (0.5 mL) as disperser solvent without salt addition. Under the selected conditions, enrichment factors up to 48-fold, limits of detection (LODs) of 5.0-10.0 ng/mL and dynamic linear ranges of 25-1500 ng/mL were obtained. A reasonable repeatability (RSD≤11.8%, n=5) with satisfactory linearity (r(2)≥0.9954) of the results illustrated a good performance of the presented method. The accuracy of the method was tested by the relative recovery experiments on spiked samples, with results ranging from 85 to 118%. Finally, the method was successfully applied for determination of the analytes in several real water samples. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Quantitative analysis of nicotinic acid,nicotinamide and 3-cyanopyridine in industrial effluent by high performance liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    TSANG Po Keung; ZENG Lixuan

    2007-01-01

    A rapid and reliable high performance liquid chromatographic(HPLC) method for the simultaneous determination of heterocyelic compounds,namely nicotinic acid,nicotinamide,and 3-cyanopyridine,in industrial effluent is described.A Φ4.6 mmx 150 mm,5 μm C-18 reversed phase stationery phase,and a methanol-acetonitrile-water tertiary mobile phase (20:20:60v/v) were used for separation.The detection wavelength of a diode array (DAD) was set at 216 nm with a bandwidth of 16 nm.Phenol was used as an internal standard.The regression equations revealed a linear relationship between the concentration of the analytes injected and the peak area detected by DAD.The limits of detection (S/N = 3) ranged from 0.70 to 1.18 mg L-1,the recoveries ranged from 87% to 102% and the precision expressed as % RSD intra-day and inter-day varied from 0.9 to 3.9 and 1.2 to 5.6,respectively.This method is rapid,sensitive and suitable for the monitoring of nicotinic acid,nicotinamide,and 3-cyanopyridine in effluent of related pharmaceutical manufacturing plants.

  11. Metabolite Analysis of Toosendanin by an Ultra-High Performance Liquid Chromatography-Quadrupole-Time of Flight Mass Spectrometry Technique

    Directory of Open Access Journals (Sweden)

    Na Li

    2013-09-01

    Full Text Available Toosendanin is the major bioactive component of Melia toosendan Sieb. et Zucc., which is traditionally used for treatment of abdominal pain and as an insecticide. Previous studies reported that toosendanin possesses hepatotoxicity, but the mechanism remains unknown. Its bioavailability in rats is low, which indicates the hepatotoxicity might be induced by its metabolites. In this connection, in the current study, we examined the metabolites obtained by incubating toosendanin with human live microsomes, and then six of these metabolites (M1–M6 were identified for the first time by ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry (UHPLC-Q-TOF/MS. Further analysis on the MS spectra showed M1, M2, and M3 are oxidative products and M6 is a dehydrogenation product, while M4 and M5 are oxidative and dehydrogenation products of toosendanin. Moreover, their possible structures were deduced from the MS/MS spectral features. Quantitative analysis demonstrated that M1-M5 levels rapidly increased and reached a plateau at 30 min, while M6 rapidly reached a maximal level at 20 min and then decreased slowly afterwards. These findings have provided valuable data not only for understanding the metabolic fate of toosendanin in liver microsomes, but also for elucidating the possible molecular mechanism of its hepatotoxicity.

  12. 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)

  13. Simultaneous determination of tolperisone and lidocaine by high performance liquid chromatography.

    Science.gov (United States)

    Liawruangrath, S; Liawruangrath, B; Pibool, P

    2001-12-01

    A reversed phase high performance liquid chromatographic (RP-HPLC) method for the simultaneous determination of tolperisone (TP) and lidocaine (LD) has been developed. The drugs were separated on a column (4.60 x 250 mm(2)) Spherisorb ODS (5 microm) using 5.5% triethylamine in 70/30 v/v acetonitrile/water as mobile phase 0.7 ml min(-1)and UV detection at 254 nm. The detection limits for Tolperisone hydrochloride (TP-HCl) and lidocaine hydrochloride (LD-HCl) were 0.20 ng/20 microl and 100 ng/20 microl and the quantitation limits were 0.50 ng/20 microl and 250 ng/20 microl, respectively. Linear calibration curves over the ranges of 1-10, 10-100 and 150-500 microg ml(-1) for TP-HCl and 10-500 microg ml(-1) for LD-HCl were established. Different calibration slopes were found for TP probably owing to changes in refractive index due to increase in TP concentration. The average recoveries of the added TP in the samples (TP-HCl tablets and injection liquid). A solutions spiked with standard TP-HCl were 99.9 and 99.7% with the RSD (n=11) of 0.66 and 0.67%, respectively. The average recovery of the added LD in the sample (injection) spiked with standard LD-HCl was 98.9% with the RSD (n=11) of 0.59%. The proposed method has been applied to the determination of TP-HCl and LD-HCl in commercial products available in Thailand. Comparative determination of TP by UV spectrophotometry and LD by colorimetry were also carried out. The results obtained by both methods were in good agreement of those obtained by the proposed method verified by using t-test. The proposed RP-HPLC method is simple, accurate, reproducible and suitable for routine analysis.

  14. [Determination of polyphenols in tobacco by solid phase extraction and high performance liquid chromatography].

    Science.gov (United States)

    Wang, L; Fang, R B; Li, Z; Jiang, C Q; Yang, G Y

    2001-11-01

    A high performance liquid chromatographic (HPLC) method for the determination of polyphenols in tobacco was studied. Polyphenols can be extracted from tobacco sample by refluxing with 80% (volume fraction) methanol, then subjected to degrease by solid phase extraction with Sep-Pak-C18 column. Chlorogenic acid, rutin, scopoletin, caffeic acid, scopolin and other polyphenols can be separated on a Nova-Pak-C18 column (3.9 mm i.d. x 150 mm) gradiently eluted with methanol and 0.05 mol.L-1 potassium dihydrogen phosphate buffer solution as mobile phase at a flow-rate of 0.5 mL.min-1. Polyphenols were determined at the maximum wavelength of each polyphenol, for chlorogenic acid 326.1 nm, rutin 354.8 nm, scopoletin 344.0 nm, caffeic acid 323.7 nm and scopolin 365.2 nm by photodiode array detector. The main polyphenols in tobacco can be identified with UV spectroscopy. The recoveries of tobacco polyphenols were 94%-105%, and the relative standard deviations were 1.28%-1.49%. This method can be applied to the determination of polyphenols in tobacco with satisfactory results.

  15. 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.

  16. Determination of 5-methylcytosine from plant DNA by high-performance liquid chromatography.

    Science.gov (United States)

    Wagner, I; Capesius, I

    1981-06-26

    The relative amounts of the five nucleosides (deoxycytidine, 5-methyldeoxycytidine, deoxyadenosine, deoxyguanosine and thymidine) in the DNA of nine plant species, one plant satellite DNA, and one animal species were determined by high performance liquid chromatography. The method allows the clean separation of the nucleosides from 10 microgram samples with 15 min. The following values for the proportion of methylated cytosines among all cytosines were obtained: Lobularia maritima 18.5%, Nicotiana tabacum 32.6%, Pisum sativum 23.2%, Rhinanthus minor 29.2%, Sinapsis alba 12.2%, Vicia faba 30.5%, Viscum album 23.2%, Cymbidium pumilum 18.8%, Cymbidium pumilum AT-rich satellite DNA 15.8%, Triticum aestivum 22.4%. DNA of an animal, the gerbil, Meriones unguiculatus, had a methylation percentage of 3.1%. An estimate of the GC content based on the buoyant density of DNA tends to be lower than the actual value, an estimate based on the melting temperature tends to be higher. This supports the finding by other authors that DNA methylation decreases the buoyant density and may increase the melting temperature at high m5C concentration.

  17. Reversed-phase high-performance liquid chromatography of unsubstituted aminobenzoic acids

    Science.gov (United States)

    Abidi, S.L.

    1989-01-01

    High-performance liquid chromatographic (HPLC) characteristics of three position isomers of aminobenzoic acids (potential metabolites of important anesthetic drugs), were delineated with respect to their interactions with various mobile phases and stationary phases. HPLC with five hydrocarbonaceous phase, I?-cyclodextrin silica (CDS), macrophase MP-1 polymer (MP), macroporous polystyrene/divinylbenzene (MPD), octadecylsilica (ODS), and propylphenylsilica (PPS), yielded results explicable in terms of substituent effects derived from the bifunctional amino- and carboxy groups. For cases where mobile phases contained sulfonates or quaternary ammonium salts both having longer chain alkyls, retention of analytes on all but CDS appeared to proceed predominantly via an ion-pairing mechanism. The extent of the corresponding counter-ion effects decreased in the order: MPD > ODS > PPS > MP, while the analyte retention order paralleled thier pH2 values. On the other hand, an inverse relationship between the magnitude of capacity factors (k') and pK1 values of the title compounds was observed in experiments that produced retention data incompatible with ion-pair interaction rationales. The unique HPLC results obtained with the CDS phase are compared with those obtained with other phases.

  18. Sources of Variability in Chlorophyll Analysis by Fluorometry and by High Performance Liquid Chromatography. Chapter 22

    Science.gov (United States)

    VanHeukelem, Laurie; Thomas, Crystal S.; Glibert, Patricia M.

    2001-01-01

    The need for accurate determination of chlorophyll a (chl a) is of interest for numerous reasons. From the need for ground-truth data for remote sensing to pigment detection for laboratory experimentation, it is essential to know the accuracy of the analyses and the factors potentially contributing to variability and error. Numerous methods and instrument techniques are currently employed in the analyses of chl a. These methods range from spectrophotometric quantification, to fluorometric analysis and determination by high performance liquid chromatography. Even within the application of HPLC techniques, methods vary. Here we provide the results of a comparison among methods and provide some guidance for improving the accuracy of these analyses. These results are based on a round-robin conducted among numerous investigators, including several in the Sensor Intercomparison and Merger for Biological and Interdisciplinary Oceanic Studies (SIMBIOS) and HyCODE Programs. Our purpose here is not to present the full results of the laboratory intercalibration; those results will be presented elsewhere. Rather, here we highlight some of the major factors that may contribute to the variability observed. Specifically, we aim to assess the comparability of chl a analyses performed by fluorometry and HPLC, and we identify several factors in the analyses which may contribute disproportionately to this variability.

  19. 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.

  20. Chiral-phase high-performance liquid chromatography of rotenoid racemates

    Science.gov (United States)

    Abidi, S.L.

    1987-01-01

    The high-performance liquid chromatograhic (HPLC) behavior of parent rotenoids (type I) and the hydroxyl-analogues (type II) on three different chiral stationary phases (CSPs) was studied. Separations of optical isomers were achieved in various degrees depending largely upon the rotenoidal structures and the CSP types employed. Enantiomers of all but elliptone compounds were separable on β-cyclodextrin-bonded silica (CDS). Without exception, the 12a-hydroxyrotenoid antipodes were resolved on Pirkle's phenylglycine-bonded silica (PGS) despite unsuccessful attenmpts to resolve the type I rotenoidal racemates. Conversely, optical resolution of the latter rotenoids was accomplished by using a helical polytriphenylmethylacrylate-coated silica (TPS) column and the observed separation factors (α values) ranged from 1.14 to 1.90. The results from HPLC of type II rotenoids on TPS (α = 1.00–1.63) suggested that variations in E-ring structures had profound influence on the resolution outcome. Conjugated double bonds on the E-ring and the desisopropylation of the five-membered E-ring ot type II rotenoids appeared to be important structural features for chiral recognition involving the TPS substrate. In both reversed-phase (CDS) and normal-pahse (PGS and TPS) HPLC modes, the less polar enantiomers were the 6aβ,12aβ-rotenoids as observed in most cases, though this relationship was reversed in the cases of deguelin and hydroxyelliptone probably due to conformational effects of rotenoidal ring systems.

  1. Detection of HEMA in self-etching adhesive systems with high performance liquid chromatography

    Science.gov (United States)

    Panduric, V.; Tarle, Z.; Hameršak, Z.; Stipetić, I.; Matosevic, D.; Negovetić-Mandić, V.; Prskalo, K.

    2009-04-01

    One of the factors that can decrease hydrolytic stability of self-etching adhesive systems (SEAS) is 2-hydroxymethylmethacrylate (HEMA). Due to hydrolytic instability of acidic methacrylate monomers in SEAS, HEMA can be present even if the manufacturer did not include it in original composition. The aim of the study was to determine the presence of HEMA because of decomposition by hydrolysis of methacrylates during storage, resulting with loss of adhesion strength to hard dental tissues of the tooth crown. Three most commonly used SEAS were tested: AdheSE ONE, G-Bond and iBond under different storage conditions. High performance liquid chromatography analysis was performed on a Nucleosil C 18-100 5 μm (250 × 4.6 mm) column, Knauer K-501 pumps and Wellchrom DAD K-2700 detector at 215 nm. Data were collected and processed by EuroCrom 2000 HPLC software. Calibration curves were made related eluted peak area to known concentrations of HEMA (purchased from Fluka). The elution time for HEMA is 12.25 min at flow rate 1.0 ml/min. Obtained results indicate that no HEMA was present in AdheSE ONE because methacrylates are substituted with methacrylamides that seem to be more stable under acidic aqueous conditions. In all other adhesive systems HEMA was detected.

  2. Determination of Decabrominated Diphenyl Ether in Soils by Soxhlet Extraction and High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Xing-Jian Yang

    2013-01-01

    Full Text Available This study described the development of a method based on soxhlet extraction combining high performance liquid chromatography (soxhlet-HPLC for the accurate detection of BDE-209 in soils. The solvent effect of working standard solutions in HPLC was discussed. Results showed that 1 : 1 of methanol and acetone was the optimal condition which could totally dissolve the BDE-209 in environmental samples and avoid the decrease of the peak area and the peak deformation difference of BDE-209 in HPLC. The preliminary experiment was conducted on the configured grassland (1 μg/g to validate the method feasibility. The method produced reliable reproducibility, simulated soils (n=4 RSD 1.0%, and was further verified by the analysis e-waste contaminated soils, RSD range 5.9–11.4%. The contamination level of BDE-209 in burning site was consistent with the previous study of Longtang town but lower than Guiyu town, and higher concentration of BDE-209 in paddy field mainly resulted from the long-standing disassembling area nearby. This accurate and fast method was successfully developed to extract and analyze BDE-209 in soil samples, showing its potential use for replacing GC to determinate BDE-209 in soil samples.

  3. 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.

  4. Description and validation of coupling high performance liquid chromatography with resonance Rayleigh scattering in aminoglycosides determination

    Energy Technology Data Exchange (ETDEWEB)

    Zhang Lei [Chemistry and Chemical Engineering, Southwest University, 400715 Chongqing (China); Peng Jingdong, E-mail: hxpengjd@swu.edu.cn [Chemistry and Chemical Engineering, Southwest University, 400715 Chongqing (China); Tang Jinxia; Yuan Binfang; He Rongxing; Xiao Ying [Chemistry and Chemical Engineering, Southwest University, 400715 Chongqing (China)

    2011-11-14

    Graphical abstract: Theoretical and experimental analysis had proved that aminoglycosides reacted with Congo red to form binary compounds simultaneously, which led to a novel HPLC-RRS strategy being applied in substances which are not fluorescing and not UV absorbed. Highlights: {yields} A novel HPLC-RRS strategy was shown in this study. {yields} Theoretical and experimental analysis had proved the feasibility of this method. {yields} Because of its specificity, no interference from the matrix was observed. {yields} The analytes in biological matrix were all well resolved without any interference. {yields} It provided new insights for analytes lack of useful spectroscopic and electrochemical properties. - Abstract: In view of the fact that many substances generally exhibit very little ultraviolet absorbance and the absence of native fluorescence, a new strategy with simple instrumentation and excellent analytical performance combining high performance liquid chromatography (HPLC) with resonance Rayleigh scattering (RRS) was developed. It was validated for the quantification of aminoglycosides (AGs). This fact was also carefully calculated by quantum chemistry. However, the sensitivity was probably limited by the volume of flow-through cell. Therefore, the result calls for a suitable one to ensure optimal RRS signal. Interestingly, when serum or urine samples of analytes were analyzed by this method, they were all well resolved without any interference, which would hold a new perspective to be applied in the determination of substances in biological matrix.

  5. The MAREDAT global database of high performance liquid chromatography marine pigment measurements

    Science.gov (United States)

    Peloquin, J.; Swan, C.; Gruber, N.; Vogt, M.; Claustre, H.; Ras, J.; Uitz, J.; Barlow, R.; Behrenfeld, M.; Bidigare, R.; Dierssen, H.; Ditullio, G.; Fernandez, E.; Gallienne, C.; Gibb, S.; Goericke, R.; Harding, L.; Head, E.; Holligan, P.; Hooker, S.; Karl, D.; Landry, M.; Letelier, R.; Llewellyn, C. A.; Lomas, M.; Lucas, M.; Mannino, A.; Marty, J.-C.; Mitchell, B. G.; Muller-Karger, F.; Nelson, N.; O'Brien, C.; Prezelin, B.; Repeta, D.; Smith, W. O., Jr.; Smythe-Wright, D.; Stumpf, R.; Subramaniam, A.; Suzuki, K.; Trees, C.; Vernet, M.; Wasmund, N.; Wright, S.

    2013-03-01

    A global pigment database consisting of 35 634 pigment suites measured by high performance liquid chromatography was assembled in support of the MARine Ecosytem DATa (MAREDAT) initiative. These data originate from 136 field surveys within the global ocean, were solicited from investigators and databases, compiled, and then quality controlled. Nearly one quarter of the data originates from the Laboratoire d'Océanographie de Villefranche (LOV), with an additional 17% and 19% stemming from the US JGOFS and LTER programs, respectively. The MAREDAT pigment database provides high quality measurements of the major taxonomic pigments including chlorophylls a and b, 19'-butanoyloxyfucoxanthin, 19'-hexanoyloxyfucoxanthin, alloxanthin, divinyl chlorophyll a, fucoxanthin, lutein, peridinin, prasinoxanthin, violaxanthin and zeaxanthin, which may be used in varying combinations to estimate phytoplankton community composition. Quality control measures consisted of flagging samples that had a total chlorophyll a concentration of zero, had fewer than four reported accessory pigments, or exceeded two standard deviations of the log-linear regression of total chlorophyll a with total accessory pigment concentrations. We anticipate the MAREDAT pigment database to be of use in the marine ecology, remote sensing and ecological modeling communities, where it will support model validation and advance our global perspective on marine biodiversity. The original dataset together with quality control flags as well as the gridded MAREDAT pigment data may be downloaded from PANGAEA: http://doi.pangaea.de/10.1594/PANGAEA.793246.

  6. Determination of Capsaicin and Dihydrocapsaicin in Capsicum Fruit Samples using High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Ayman Abdel Ghafar

    2011-10-01

    Full Text Available The aim of the present study was to determine the content of capsaicin and dihydrocapsaicin in Capsicum samples collected from city markets in Riyadh (Saudi Arabia, calculate their pungency in Scoville heat units (SHU and evaluate the average daily intake of capsaicin for the population of Riyadh. The investigated samples consisted of hot chillies, red chillies, green chillies, green peppers, red peppers and yellow peppers. Extraction of capsaicinoids was done using ethanol as solvent, while high performance liquid chromatography (HPLC was used for separation, identification and quantitation of the components. The limit of detection (LOD of the method was 0.09 and 0.10 µg/g for capsaicin and dihydrocapsaicin, respectively, while the limit of quantification (LOQ was 0.30 and 0.36 µg/g for capsaicin and dihydrocapsaicin, respectively. Hot chillies showed the highest concentration of capsaicin (4249.0 ± 190.3 µg/g and the highest pungency level (67984.60 SHU, whereas green peppers had the lowest detected concentration (1.0 ± 0.9 µg/g; green peppers, red peppers and yellow peppers were non pungent. The mean consumption of peppers for Riyadh city population was determined to be 15.5 g/person/day while the daily capsaicin intake was 7.584 mg/person/day.

  7. 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

  8. AB063. Prevalence of thalassemias and hemoglobinopathies detected via high performance liquid chromatography in Filipinos

    Science.gov (United States)

    Silao, Catherine Lynn; Fabella, Terence Diane; Yuson, Ernesto; Naranjo, Maria Liza; Padilla, Carmencita

    2015-01-01

    Background The thalassemias and hemoglobinopathies are groups of autosomal recessive inherited blood disorders affecting the quantity and quality of hemoglobin. Patients with the disease can vary from clinically asymptomatic to transfusion dependent individuals. It is considered prevalent in many parts of the world particularly in Southeast Asia. Thus, screening and prevalence determination of these diseases are important in the Filipino population. The study aims to determine the prevalence of thalassemias and hemoglobinopathies in the Philippines using high performance liquid chromatography. Methods Referred patients by hematologists from different parts of the country from October 2008 to December 2014 were included. Peripheral blood extracted from the subjects, were hemolyzed and screened for thalassemias and hemoglobinopathies using VariantTM HPLC. Data interpretations were based on levels of the HbA2 Fetal Hb and HbA detected. Results Majority of the patients were beta thalassemics followed by alpha-thalassemics. Hemoglobin E was found in 1% of the population tested while 2% of the patients have beta thalassemia with HbE interaction. Conclusions A significant proportion of thalassemias and hemoglobinopathies were determined from the 6-year screening of Filipinos using VariantTM HPLC. The results of this study provide not only confirmation of the occurrence and prevalence of these growing and diverse groups of genetic blood diseases but also suggest that the use of HPLC is a useful screening tool.

  9. 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.

  10. Determination of sulphachloropyrazine-diaveridine residues by high performance liquid chromatography in broiler edible tissues

    Science.gov (United States)

    LI, Yongjun; BU, Shijin

    2015-01-01

    Diaveridine (DVD) is used in combination with sulphachloropyrazine (SPZ) as an effective antibacterial agent and antiprotozoal agent, respectively, in humans and animals. To gain a better understanding of the metabolism of SPZ and DVD in the food-producing animals, a high performance liquid chromatography (HPLC) method to determine and quantify sulphachloropyrazine (SPZ) and diaveridine (DVD) suspension residues from broilers is reported. Thirty healthy chickens were orally administered with sulphachloropyrazine-diaveridine (SPZ-DVD) suspension in water of 300 mg/l (SPZ) per day for seven successive days. Six chickens per day were slaughtered at 0, 1, 3, 5 and 7 days after the last administration. This procedure permitted SPZ and DVD to be separated from muscle tissue, liver, kidneys and skin with fat after extraction with acetonitrile and acetone under slightly acidic conditions. From the detected residuals in different tissues, we found that SPZ was quickly eliminated in liver and muscle, and slowly eliminated in kidney and skin with fat. DVD was quickly eliminated in liver and slowly eliminated in kidney. The withdrawal period of SPZ was 3.26, 3.72, 4.39 and 5.43 days in muscle, liver, kidney and skin with fat, respectively. The withdrawal period of DVD was 4.77, 4.94, 6.74 and 4.58 days in muscle, liver, kidney and skin with fat, respectively. Therefore, the suggested withdrawal period for SPZ-DVD suspension should be 7 days after dosing for seven successive days. PMID:26212255

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

    Science.gov (United States)

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

    2013-04-01

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

  12. Determination of multiple human arsenic metabolites employing high performance liquid chromatography inductively coupled plasma mass spectrometry

    Science.gov (United States)

    Stice, Szabina; Liu, Guangliang; Matulis, Shannon; Boise, Lawrence H.; Cai, Yong

    2016-01-01

    During the metabolism of different arsenic-containing compounds in human, a variety of metabolites are produced with significantly varying toxicities. Currently available analytical methods can only detect a limited number of human metabolites in biological samples during one run due to their diverse characteristics. In addition, co-elution of species is often unnoticeable with most detection techniques leading to inaccurate metabolic profiles and assessment of toxicity. A high performance liquid chromatography inductively coupled mass spectrometry (HPLC-ICP-MS) method was developed that can identify thirteen common arsenic metabolites possibly present in human with special attention dedicated to thiolated or thiol conjugated arsenicals. The thirteen species included in this study are arsenite (AsIII), arsino-glutathione (As(GS)3), arsenate (AsV), monomethylarsonous acid (MMAIII), monomethylarsino-glutathione (MMAIII(GS)2), monomethylarsonic acid (MMAV), dimethylarsinous acid (DMAIII (from DMAIIII)), S-(dimethylarsinic)cysteine (DMAIII(Cys)), dimethylarsino-glutathione (DMAIII(GS)), dimethylarsinic acid (DMAV), dimethylmonothioarsinic acid (DMMTAV), dimethyldithioarsinic acid (DMDTAV), dimethylarsinothioyl glutathione (DMMTAV(GS)). The developed method was applied for the analysis of cancer cells that were incubated with Darinaparsin (DMAIII(GS)), a novel chemotherapeutic agent for refractory malignancies, and the arsenic metabolic profile obtained was compared to results using a previously developed method. This method provides a useful analytical tool which is much needed in unequivocally identifying the arsenicals formed during the metabolism of environmental arsenic exposure or therapeutic arsenic administration. PMID:26708625

  13. 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.

  14. Analysis of lipoprotein profiles of healthy cats by gel-permeation high-performance liquid chromatography.

    Science.gov (United States)

    Mizutani, Hisashi; Sako, Toshinori; Okuda, Hiroko; Arai, Nobuaki; Kuriyama, Koji; Mori, Akihiro; Yoshimura, Itaru; Koyama, Hidekazu

    2016-09-01

    Density gradient ultracentrifugation (DGUC) and gel electrophoresis are conventionally used to obtain lipoprotein profiles of animals. We recently applied high-performance liquid chromatography with a gel permeation column (GP-HPLC) and an on-line dual enzymatic system to dogs for lipoprotein profile analysis. We compared the GP-HPLC with DGUC as a method to obtain a feline lipoprotein profile. The lipoprotein profiles showed large and small peaks, which corresponded to high-density lipoprotein (HDL) and low-density lipoprotein (LDL), respectively, whereas very low-density lipoprotein (VLDL) and chylomicron (CM) were only marginally detected. This profile was very similar to that of dogs reported previously. Healthy cats also had a small amount of cholesterol-rich particles distinct from the normal LDL or HDL profile. There was no difference in lipoprotein profiles between the sexes, but males had a significantly larger LDL particle size (P=0.015). This study shows the feasibility of GP-HPLC for obtaining accurate lipoprotein profiles with small sample volumes and provides valuable reference data for healthy cats that should facilitate diagnoses.

  15. Quantification of mRNA by RT-competitive-PCR and high performance liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The use of RT-competitive-PCR with high performance liquid chromatography (HPLC) detection to quantify the absolute number of mRNA copies in mammalian cells is reported. As an example, the glutathione transferase (GST)-α mRNA in human hepatoma Hep G2 cells has been estimated. A PCR-generated internal standard was used as a competitor, co-amplified with the GST-α target sequence. The RT-competitive-PCR method was improved by designing target and competitor molecules which differed in only 30 base pairs. This allowed the two sequences to be co-amplified with the same efficiency. This improvement also facilitated a wider ratio to be used than previous methods (target:competitor ratio between 0.2 and 5). Products were baseline separated by HPLC using an ion-exchange column readily quantified at 260 nm. To validate the improved methodology, the effect of a known GST-α inducer, the anticancer drug oltipraz, was shown to induce GST-α mRNA up to 3-fold in Hep G2 cells. The RT-competitive PCR-HPLC method provides a reliable and sensitive way to quantify the amount of specific mRNA with 0.1 ng of total RNA.

  16. DETERMINATION OF TELMISARTAN IN PHARMACEUTICAL FORMULATIONS BY REVERSE PHASE-HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

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    Manoj S. Charde

    2012-05-01

    Full Text Available A simple, sensitive and reproducible reverse-phase high performance liquid chromatographic (RP-HPLC method has been developed for the quantitative estimation of Telmisartan (TELM in the pharmaceutical formulations. Chromatographic separation was achieved on a 250 × 4.6 mm, 5μ, Waters symmetry column. The flow rate was 1 ml/min and eluent was monitored by absorbance at 230 nm using a mixture of Methanol and Acetonitrile (pH 3.0±0.01 in the ratio of 30:70 (v/v. The retention time of Telmisartan was found to be 7.9 min. Calibration plots were linear in the concentration range of 10-50 μg/ml for Telmisartan with correlation coefficient (R2 0.999. The proposed method was validated by testing its linearity, recovery, specificity, system suitability, precision (Interday, intraday, analyst and instrument precision, robustness and LOD/LOQ values and it was successfully employed for the determination of Telmisartan in pharmaceutical tablet formulations.

  17. Determination of Acrylamide Metabolite ,Mercapturic Aicd by High Performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    WUYi-Qun; YUAi-Ru; 等

    1993-01-01

    A HPLC Assay was developed to indentify and measure the metabolite of acrylamide,mercapturic acid,N-Acetyl-s-(propionamide)-cyteine(APC)in urine O-phthalaldehyde (OPA)was utilized as a precolumm derivatizing agent in the assay.This derivative was isolated with a good selectivity by high performance liquid chromatography(HPLC)employing reversed phase ODS columns.The quantiation of the mercapturic acid derivative was reproducible and with a detection limit of 1 pmol.The average coefficient of variation for the runs carried out on the same day was approximately 4.6% at the range of 80-160μmol·l-1 of APC,and the mean analytical recovery from urine samples was 94%.The metabolite of urine of workers exposed to acrylamide was identified as N-acetyl-s-(propionamide)-cysteine by HPLC chromatography and fluorescence scan and HPLC-Mass spectra.All results were identical with the authentic synthesized compound.

  18. [Determination of histamine in canned fish by high performance liquid chromatography with pre-column derivatization].

    Science.gov (United States)

    Jin, Gaowa; Cai, Youqiong; Yu, Huijuan; Qian, Beilei

    2010-11-01

    A pre-column derivatization-high performance liquid chromatographic (HPLC) method has been developed for the determination of histamine in canned fish. The homogenated samples were ultrasonically extracted with perchloric acid aqueous solution, derivatized with dansyl chloride and diluted with acetonitrile to a desired volume. The samples were determined by HPLC with ultraviolet detector and quantified by external standard method. Adopting a C18 column with 1.8 microm stationary phase particles, the analysis time for each sample was smaller than 5 min with the flow rate of 0.3 mL/min. It can decrease the consumption of the mobile phase and save the cost. The linear range was 0.08-8.00 mg/L for histamine. The correlation coefficient was 0.999 98. The average recoveries of histamine at different concentration levels in spiked samples were greater than 96% and the relative standard deviations (RSDs) were smaller than 2.5%. The quantitation limit was 5.00 mg/kg for histamine in canned fish by HPLC. The results indicated that this HPLC method is fast, sensitive, reproducible and practical for the routine analysis of histamine in canned fish.

  19. A stability-indicating high performance liquid chromatography method to determine apocynin in nanoparticles

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    Juliana Kovalczuk de Oliveira

    2017-04-01

    Full Text Available In this study, we developed and validated a fast, specific, sensitive, precise and stability-indicating high performance liquid chromatography (HPLC method to determine the drug apocynin in bovine serum albumin (BSA nanoparticles. Chromatographic analyses were performed on an RP C18 column and using a photodiode array detector at a wavelength of 276 nm. Mobile phase consisted of a mixture of acetonitrile and 1% acetic acid (60:40, v/v, and it was eluted isocratically at a flow rate of 0.8 mL/min. The retention time of apocynin chromatographic peak was 1.65 min. The method was linear, precise, accurate and specific in the range of 5–100 μg/mL. The intra- and inter-day precisions presented relative standard deviation (RSD values lower than 2%. The method was robust regarding changes in mobile phase proportion, but not for flow rate. Limits of detection and quantitation were 78 ng/mL and 238 ng/mL, respectively. Apocynin was exposed to acid and alkali hydrolysis, oxidation and visible light. The drug suffered mild degradation under acid and oxidation conditions and great degradation under alkali conditions. Light exposure did not degrade the drug. The method was successfully applied to determine the encapsulation efficiency of apocynin in BSA nanoparticles.

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

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    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.

  1. High-performance liquid chromatographic separation of subcomponents of antimycin A.

    Science.gov (United States)

    Abidi, S L

    1988-08-05

    Using a reversed-phase high-performance liquid chromatographic (HPLC) technique, a mixture of antimycins A was separated into eight hitherto unreported subcomponents, A1a, A1b, A2a, A2b, A3a, A3b, A4a, and A4b. Although a base-line resolution of the known four major antimycins A1, 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 modifers, 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 interpreted 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.

  2. High-performance liquid-chromatographic separation of subcomponents of antimycin-A

    Science.gov (United States)

    Abidi, S.L.

    1988-01-01

    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.

  3. High-Performance Liquid Chromatographic Quantification of Flavonoids in Eriocaulaceae Species and Their Antimicrobial Activity

    Directory of Open Access Journals (Sweden)

    Lourdes Campaner dos Santos

    2009-11-01

    Full Text Available Quantification of prepared samples by analysis using high performance liquid chromatography with DAD detection was developed to analyze rutin, 6-methoxyapigenin, and 6-methoxyapigenin-7-O-β-D-glucopyranoside isolated from methanolic extracts of different genus: Syngonanthus, Leiothix and Eriocaulon (Eriocaulaceae. The linearity, accuracy, and the inter-day precision of the procedure were evaluated. The calibration curves were linear. The recoveries of the flavonoids in the samples analyzed were 96.3% to 98.5%. The percentage coefficient of variation for the quantitative analysis of the flavonoids in the analyses of the samples was under 5%. The antimicrobial activity of the five methanol extracts of these Eriocaulaceae species was assayed against the microorganisms Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Salmonella setubal, Saccharomyces cerevisiae and Candida albicans. Measured MIC values ranged from 1.25 to 10.00 mg/mL. The flavonoid contents suggest that Eriocaulaceae species may be a promising source of compounds to produce natural phytomedicines.

  4. 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.

  5. High-Performance Liquid Chromatographic Resolution of Neutral and Cationic Hetero[6]Helicenes.

    Science.gov (United States)

    Labrador, Geraldine M; Bosson, Johann; Breitbach, Zachary S; Lim, Yeeun; Francotte, Eric R; Sabia, Rocchina; Villani, Claudio; Armstrong, Daniel W; Lacour, Jérôme

    2016-04-01

    Cationic hetero[6]helicenes 1+, 2+ and 3+ have been recently disclosed. Herein we report on their enantiomeric separation using high-performance liquid chromatography. Separation of the antipodes can be achieved in preparative scale on neutral adducts with Chiralcel OD-I or Chiralpak ID CSP. Selectivity factors of 1.90, 1.67, and 1.96 were obtained for 1-H, 2-H, and 3-H, respectively. Separation can also be performed on the carbenium ions on regular Chiralpak IA CSP using water-containing eluents, thus allowing for enantiomeric purity determinations in aqueous environments. Resolution of neutral and cationic helicenes is also achieved on more recently developed LARIHC columns. The versatility of the cyclofructan phases allows for baseline separations for both cases and their loading capabilities are demonstrated. Finally, the configurational stability of 1+, 2+, and 3+ was measured. For each replacement of an oxygen atom by an amino group, the racemization barrier increases significantly (ΔG‡ = 29.8, 36.3 and >37 kcal mol(-1) for 1+, 2+, and 3+ respectively).

  6. Development and Validation of Reversed-Phase High Performance Liquid Chromatographic Method for Hydroxychloroquine Sulphate.

    Science.gov (United States)

    Singh, A; Roopkishora; Singh, C L; Gupta, R; Kumar, S; Kumar, M

    2015-01-01

    In the present work new, simple reversed-phase high performance liquid chromatographic method was developed and validated for the determination of hydroxychloroquine sulphate in blood plasma. Chloroquine sulphate was used as an internal standard. The chromatographic separation was achieved with octadecyl silane Hypersil C18 column (250×6 mm, 5 μm) using water and organic (acetonitrile:methanol: 50:50, v/v) mobile phase in 75:25 v/v ratio, with sodium 1-pentanesulfonate and phosphoric acid. This organic phase was maintained at pH 3.0 by orthophosphoric acid. The flow rate of 2.0 ml/min(.) with detection at 343 nm was used in the analysis. The calibration curve of standard hydroxychloroquine sulphate was linear in range 0.1-20.0 μg/ml. The method was validated with respected to linearity, range, precision, accuracy, specificity and robustness studies according to ICH guidelines. The method was found to be accurate and robust to analyze the hydroxychloroquine sulphate in plasma samples.

  7. Development and validation of a high-performance liquid chromatographic method for bioanalytical application with rimonabant.

    Science.gov (United States)

    Bhaumik, Uttam; Ghosh, Animesh; Chatterjee, Bappaditya; Sengupta, Pinaki; Darbar, Soumendra; Roy, Bikash; Nandi, Utpal; Pal, Tapan K

    2009-05-01

    A simple and feasible high-performance liquid chromatographic method with UV detection was developed and validated for the quantification of rimonabant in human plasma. The chromatographic separation was carried out in a Hypersil BDS, C(18) column (250 mm x 4.6mm; 5 microm). The mobile phase was a mixture of 10 mM phosphate buffer and acetonitrile (30:70, v/v) at a flow rate of 1.0 ml/min. The UV detection was set at 220 nm. The extraction recovery of rimonabant in plasma at three quality control (QC) samples was ranged from 84.17% to 92.64%. The calibration curve was linear for the concentration range of 20-400 ng/ml with the correlation coefficient (r(2)) above 0.9921. The method was specific and sensitive with the limit of quantification of 20 ng/ml. The accuracy and precision values obtained from six different sets of QC samples analyzed in separate occasions ranged from 88.13% to 106.48% and 0.13% to 3.61%, respectively. In stability tests, rimonabant in human plasma was stable during storage and assay procedure. The method is very simple, sensitive and economical and the assay was applied to human plasma samples in a clinical (pharmacokinetic) study of rimonabant.

  8. Determination of purine contents in different parts of pork and beef by high performance liquid chromatography.

    Science.gov (United States)

    Rong, Shengzhong; Zou, Lina; Zhang, Yannan; Zhang, Guangteng; Li, Xiaoxia; Li, Miaojing; Yang, Fenghua; Li, Chunmei; He, Yingjuan; Guan, Hongjun; Guo, Yupeng; Wang, Dong; Cui, Xinyu; Ye, Hongting; Liu, Fenghai; Pan, Hongzhi; Yang, Yuexin

    2015-03-01

    Determination of adenine, hypoxanthine, guanine and xanthine in different parts of pork and beef using high performance liquid chromatography was described. Chromatographic separation was carried out on Waters Atlantis T3 column (4.6 mm × 250 mm × 5 μm) with column temperature at 30 °C. The mobile phase contained 99% 10.0 mmol/L ammonium formate solution at pH 3.6 and 1.0% methanol. Chromatography was achieved at a flow rate of 1.0 mL/min and detection wavelength at 254 nm. The results indicated that total purine amounts in pork rump and beef sirloin were higher than those in other parts (Ppurine bases were hypoxanthine and adenine, and hypoxanthine content was the most highest in all samples (Ppurine and uricogenic purine base, we suggest that excess consumption of them be avoid, whereas pork loin chop and beef rib eye are more suitable for a low-purine diet. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. A novel high-performance liquid chromatography fingerprint approach to discriminate Phyllostachys pubescens from China

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    Jin Yong-Chun

    2012-01-01

    Full Text Available Background: A high-performance liquid chromatography fingerprint of different variants of Phyllostachys pubescens was developed for the source discrimination and quality control of Phyllostachys pubescens medicines produced in Zhejiang Province, China. Materials and Methods: Samples of Phyllostachys pubescens collected from nine cultivars in Zhejiang Province were used to establish the fingerprint. Taking the chromatographic peak (isoorientin as the reference peak, the chromatographic column was Sunfire C 18 ODS (250 mm×4.6 mm, 5 ΅m at a column temperature of 25°C. The mobile phase was acetonitrile 0.8% acetic acid water solution (in a gradient elution mode at a flow rate of 1 ml/min. The detection wavelength was 330 nm. Results: 20 common peaks were established in the chromatographic fingerprints of Phyllostachys pubescens. And then by comparing the UV and standard substance with those of the authentic standards and literature, eight main peaks in the fingerprints were identified. Data of nine cultivars were evaluated statistically using similarity analysis, hierarchical cluster analysis (HCA in order to establish a similarity standard of fingerprint for quality control of Phyllostachys pubescens, then to classify the samples. The similarity indexes were all above 0.95 between reference chromatogram and each sample′s chromatogram. Conclusion: in this study, the methods established are reliable, and could be used to evaluate the quality and to identify the species of Phyllostachys pubescens in the future.

  10. Chemical fingerprint analysis of Gentianae Radix et Rhizoma by high-performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Baozhong Duan

    2012-02-01

    Full Text Available Gentianae Radix et Rhizoma (also called “Longdan” in Chinese is commonly used for eliminating damp-heat and quenching the fire of liver and gall bladder in traditional Chinese medicine. In this study, a novel and reliable method using high-performance liquid chromatography (HPLC was developed both for quantitative analysis of four bioactive compounds (loganic acid, swertiamarin, gentiopicroside and sweroside and chemical fingerprint analysis of “Longdan”. In quantitative analysis, four compounds showed good regressions (R2>0.9987 within the test ranges and the recovery of the method was in the range 97.61−102.49%. In fingerprint analysis, ten characteristic peaks were selected to evaluate the similarities of the crude drugs, and the HPLC chromatograms of twenty samples from different regions of China showed similar patterns. The results demonstrated that the combination of the quantitative and chromatographic fingerprint analyses offered an efficient way to evaluate the quality consistency of Gentianae Radix et Rhizoma.

  11. Fingerprint chromatogram analysis of Pseudostellaria heterophylla (Miq.) Pax root by high performance liquid chromatography.

    Science.gov (United States)

    Han, Chao; Chen, Junhui; Chen, Bo; Lee, Frank Sen-Chun; Wang, Xiaoru

    2006-09-01

    A simple and reliable high performance liquid chromatographic (HPLC) method has been developed and validated for the fingerprinting of extracts from the root of Pseudostellaria heterophylla (Miq.) Pax. HPLC with gradient elution was performed on an authentic reference standard of powdered P. heterophylla (Miq.) Pax root and 11 plant samples of the root were collected from different geographic locations. The HPLC chromatograms have been standardized through the selection and identification of reference peaks and the normalization of retention times and peak intensities of all the common peaks. The standardized HPLC fingerprints show high stability and reproducibility, and thus can be used effectively for the screening analysis or quality assessment of the root or its derived products. Similarity index calculations based on cosine angle values or correlation methods have been performed on the HPLC fingerprints. As a group, the fingerprints of the P. heterophylla (Miq.) Pax samples studied are highly correlated with closely similar fingerprints. Within the group, the samples can be further divided into subgroups based on hierarchical clustering analysis (HCA). Sample grouping based on HCA coincides nicely with those based on the geographical origins of the samples. The HPLC fingerprinting techniques thus have high potential in authentication or source-tracing types of applications.

  12. Determination of capsaicin and dihydrocapsaicin in Capsicum fruit samples using high performance liquid chromatography.

    Science.gov (United States)

    Al Othman, Zeid Abdullah; Ahmed, Yacine Badjah Hadj; Habila, Mohamed Abdelaty; Ghafar, Ayman Abdel

    2011-10-24

    The aim of the present study was to determine the content of capsaicin and dihydrocapsaicin in Capsicum samples collected from city markets in Riyadh (Saudi Arabia), calculate their pungency in Scoville heat units (SHU) and evaluate the average daily intake of capsaicin for the population of Riyadh. The investigated samples consisted of hot chillies, red chillies, green chillies, green peppers, red peppers and yellow peppers. Extraction of capsaicinoids was done using ethanol as solvent, while high performance liquid chromatography (HPLC) was used for separation, identification and quantitation of the components. The limit of detection (LOD) of the method was 0.09 and 0.10 µg/g for capsaicin and dihydrocapsaicin, respectively, while the limit of quantification (LOQ) was 0.30 and 0.36 µg/g for capsaicin and dihydrocapsaicin, respectively. Hot chillies showed the highest concentration of capsaicin (4249.0 ± 190.3 µg/g) and the highest pungency level (67984.60 SHU), whereas green peppers had the lowest detected concentration (1.0 ± 0.9 µg/g); green peppers, red peppers and yellow peppers were non pungent. The mean consumption of peppers for Riyadh city population was determined to be 15.5 g/person/day while the daily capsaicin intake was 7.584 mg/person/day.

  13. 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.

  14. Quantitative determination of eleutheroside B and E from Acanthopanax species by high performance liquid chromatography.

    Science.gov (United States)

    Kang, J S; Linh, P T; Cai, X F; Kim, H S; Lee, J J; Kim, Y H

    2001-10-01

    Reversed-phase high performance liquid chromatographic method was applied for the determination of eleutheroside B and E in the various Acanthopanax species collected in Korea. The stationary phase used was Zorbax 300 SB C18 and a mobile phase program was used, which started at 6% acetonitrile for 2 min, and then a linear gradient was operated for the next 18 min to 17% acetonitrile at a flow rate of 1.0 ml/min. The column effluent was monitored at UV 210 nm. Identification was carried out by comparing the retention time and the LC/MS spectrum of each peak corresponding to eleutheroside B and E from sample with those of standards. In general, the contents of eleutheroside B and E in stems were higher than those in roots. Acanthopanax species could be classified into two groups based upon the contents of eleutheroside B and E: one group contains no or very little eleutheroside B and another contains both eleutheroside B and E.

  15. The determination of bromazepam in plasma by reversed-phase high-performance liquid chromatography.

    Science.gov (United States)

    Le Solleu, H; Demotes-Mainard, F; Vinçon, G; Bannwarth, B

    1993-08-01

    A reversed-phase high-performance liquid chromatographic method has been developed for the determination of bromazepam, an anxiolytic benzodiazepine, in plasma. After a single-step extraction from alkalinized plasma with diethyl-ether in the presence of an internal standard (alpha-hydroxy-triazolam), the residues were chromatographed on a reversed-phase Nova Pak 5 microns C18 column, with a mobile phase of acetonitrile-water-triethylamine (700:300:4, v/v/v) adjusted to pH 7.4 with orthophosphoric acid. The limit of detection was 50 ng ml-1, using a 20 microliters injection with UV detection at 240 nm. Between-day and within-day relative standard deviations were lower than 6%. Studies of drug stability during sample storage at -20 degrees C and at +4 degrees C showed no degradation of bromazepam. However, bromazepam seemed to be degraded at ambient temperature, without any influence of light. This method is applied to the determination of bromazepam plasma levels in analytical toxicology.

  16. Development of high-performance liquid chromatographic for quality and authenticity control of Chinese propolis.

    Science.gov (United States)

    Cui-ping, Zhang; Shuai, Huang; Wen-ting, Wei; Shun, Ping; Xiao-ge, Shen; Ya-jing, Li; Fu-liang, Hu

    2014-07-01

    A RP-high-performance liquid chromatography (HPLC) method was developed for quality control of Chinese propolis by simultaneous analysis of 12 flavonoids and 8 phenolic acids. The results showed that vanillic acid, rutin, myricetin, and luteolin were not detected in all of the analyzed propolis and poplar tree gum samples. The caffeic acid, ferulic acid and p-coumaric acid were not detected in poplar tree gum but were detected in propolis, which suggest that they are practical indexes of distinguishing propolis from poplar tree gum. The flavonoid profiles of poplar tree gum were found to be similar to those of propolis, which are dominated by pinobanksin, pinocembrin, 3-O-acetylpinobanksin, chrysin, and galangin. Therefore, the proposed method could be applied to exclude poplar tree gum from propolis with cafferic acid, ferulic acid, and p-coumaric acid as qualitative markers, and distinguish poplar source resin from other illegal substances, and evaluate the quality grading of poplar-type propolis with pinobanksin, pinocembrin, 3-O-acetylpinobanksin, chrysin, and galangin as qualitative and quantitative markers. © 2014 Institute of Food Technologists®

  17. Quantitation of aflatoxins in walnut kernels by high-performance liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Kabak, Bulent

    2014-01-01

    A total of 85 walnut samples collected between October 2012 and April 2013 in different provinces of Turkey were analysed for the presence of aflatoxins (AFs). The method involved methanol-water extraction, clean-up with immunoaffinity columns and a sensitive high-performance liquid chromatography coupled with fluorescence detection after post-column derivatisation. The method was validated for selectivity, linearity, trueness, precision, limit of detection and limit of quantification (LOQ), which met the performance criteria as set by EC regulation No. 401/2006. LOQs were 0.07, 0.04, 0.09 and 0.05 µg kg(-1) for AFB1, AFB2, AFG1 and AFG2, respectively. AFs were present in 9.4% of walnut samples (8/85) at total AFs levels ranging from 0.09 to 15.4 µg kg(-1). Only one of eight walnut samples exceeded the European Union limit of 2 and 4 µg kg(-1) for AFB1 and total AFs, respectively.

  18. Determination of Benzalkonium Chloride in Nasal Drops by High-Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Danijela A. Kostić

    2012-01-01

    Full Text Available A high-performance liquid chromatography (HPLC system was used in the reversed phase mode for the determination of benzalkonium chloride (BKC in nosal drops. A Chromolit RP-18e, 100 x 4.6, (UM6077/035 column was used at 40 °C. The mobile phase, optimized through an experimental design, was a 70:30 (v/v mixture of 0.057M Na-heksansulphonate potassium, dihydrogen orthophosphate buffer (pH 2.9 and acetonitrile, pumped at a flow rate of 1.75 mL/min at maintaining column temperature at 40 °C. Maximum UV detection was achieved at 215 nm. The method was validated in terms of selectivity, linearity, repeatability, precision and accuracy. The method was successfully applied for the determination of BKC in a pharmaceutical formulation of nasal drop solution without any interference from common excipients and drug substance. All the validation parameters were within the acceptance range, concordant to ICH guidelines.

  19. Determination of benzalkonium chloride in viscous ophthalmic drops of azithromycin by high-performance liquid chromatography *

    Science.gov (United States)

    Shen, Yan; Xu, Sheng-jie; Wang, Shi-chun; Tu, Jia-sheng

    2009-01-01

    A high-performance liquid chromatography (HPLC) system was used in the reversed phase mode for the determination of benzalkonium chloride (BKC) in azithromycin viscous ophthalmic drops. A Venusil-XBP(L)-C18 (150 mm×4.6 mm, 5 μm) column was used at 50 °C. The mobile phase consisted of a mixture of methanol-potassium phosphate (16:5, v/v). Two sample preparation methods were compared. The results suggested that, compared with an extraction procedure, a deproteinization procedure was much quicker and more convenient. Using the deproteinization procedure for sample preparation, calibration curves were linear in the range 5.0~50 μg/ml. The within-day and inter-day coefficients of variation were less than 10%. The average recoveries were determined as 96.70%, 98.52%, and 97.96% at concentrations of 10.0, 30.0, and 50.0 μg/ml, respectively. Variability in precision did not exceed 5%. In conclusion, this HPLC method using a simple sample treatment procedure appears suitable for monitoring BKC content in azithromycin viscous ophthalmic drops. PMID:19946951

  20. High-performance liquid chromatographic analysis and pharmacokinetics of terazosin in healthy volunteers.

    Science.gov (United States)

    Kang, B C; Yang, C Q; Rhee, J E; Suh, O K; Shin, W G

    2001-01-01

    A high-performance liquid chromatographic (HPLC) analysis of terazosin in 1 ml of human plasma was developed using prazosin as an internal standard. The plasma sample was extracted with dichloromethane and ethylether and a 100-microl aliquot was injected onto the reversed-phase column. The mobile phase, 0.02 M sodium phosphate buffer:acetonitrile:tetrahydrofuran = 720:220:60 (v/v/v), was run at a flow rate of 0.8 ml/min and the column effluent was monitored using a florescence detector set at 370 and 250 nm for the emission and excitation wave numbers, respectively. The retention times for terazosin and prazosin were approximately 6.4 and 9.8 min, respectively, and the coefficients of variation of terazosin were generally low, below 6.4%. The present HPLC method was successful for the pharmacokinetic study of terazosin in healthy volunteers. Following oral administration of terazosin, 2 mg, to 20 healthy male volunteers, the area under the plasma concentration-time curve from time zero to time infinity was 421 +/- 71.8 ng h/ml and terminal half-life was 9.83 +/- 1.29 h.

  1. Determination of Residual Monomers in Polycarboxylate Superplasticizer Using High Performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    GUO Liping; WANG Shaofeng; ZHANG Anfu; LEI Jiaheng; DU Xiaodi

    2011-01-01

    A procedure was developed for the determination of residual monomers in polycarboxylate superplasticizer (PCs) by reversed-phase high performance liquid chromatography (RP-HPLC). Four kinds of residual monomers were well separated and determined on a SinoChrom ODS-BP (C18) column with mobile phases composed of acetonitrile and phosphate buffer solution. The monomers were detected by UV detector at 205 nm and quantitatively analyzed with an external standard method. For those residual monomers, the linear response ranged from 4.0× 10-6 mol·L-1 to 2.0× 10-3 mol·L-1. The determination limit of acrylic acid, sodium methylallyl sulfonate and 2-Acrylamido-2-methylpropane sulfonic acid was 0.02× 10-5 mol·L-1, while that of methoxy-polyethylene glycol monoacrylate was 0.1 × 10-5 mol· L-1. The relative standard deviation (RSD) of high concentration samples was less than 1%, while that of the low concentration samples was between 1%-4%. The standard (additional) recovery ratio was 97.4% -104.2%.

  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. [Determination of six pesticides in milk using cloud point extraction-high performance liquid chromatography].

    Science.gov (United States)

    Wang, Jian; Cui, Yanmei; Liu, Wei; Yang, Mingmin; Chen, Jianbo

    2007-11-01

    The feasibility of employing cloud point extraction (CPE) as extraction and preconcentration method for the recovery of herbicides from milk samples followed by high performance liquid chromatography (HPLC) analysis is demonstrated. An aqueous surfactant solution containing 60 g/L Tween 20 or Triton X-100 was heated with an appropriate concentration of (NH4)2SO4 or NaCl for the extraction of herbicides. The extract was analyzed by HPLC subsequently. Six herbicides in milk were analyzed simultaneously. The results showed that the linear dynamic ranges of detection were 20 - 10 000 microg/L for tralkoxydim, metribuzin and bromoxynil, 30 - 10 000 microg/L for mefenacet, and 50 - 10 000 microg/L for bensulfuron-methyl and nicosulfuron. The correlation coefficients were 0.998 1 - 0.999 7. The average recoveries of the six herbicides ranged from 85.09% to 96.74%. The relative standard deviations for the six herbicides were in the range of 1.90% - 3.98%. The limits of detection for the six pesticides were lower than the maximum residue limits (MRL) of China. The results indicate that the method is easier, faster, sensitive and produces less pollutants.

  4. Analysis of ciprofloxacin by a simple high-performance liquid chromatography method.

    Science.gov (United States)

    Wu, Shihn-Sheng; Chein, Chih-Yuan; Wen, Yen-Hsia

    2008-07-01

    A simple and sensitive high-performance liquid chromatographic method is described for the quantitative analysis of ciprofloxacin in pharmaceuticals and human plasma. The method employs reversed-phase chromatography using an RP-C18 column with an isocratic mobile phase of acetonitrile-2% acetic acid aqueous solution (16:84, v/v), umbelliferone as an internal standard, and a flow rate of 1.0 mL/min. The UV detector is set at 280 nm. The limit of detection is 0.25 microM (S/N = 3, injection volume = 10 microL). The regression equations are linear (r > 0.9999) over a range between 0.51 approximately 130 microM for the pharmaceutical analysis of ciprofloxacin and 0.51 approximately 64.8 microM for the biological analysis of ciprofloxacin in human plasma. The intra- and inter-day relative standard deviation and relative error are less than 3.39% and 5.71%, respectively. All the recoveries are greater than 93.8%. This method is successfully applied in a pharmacokinetic study of a volunteer who receives a 500 mg ciprofloxacin tablet.

  5. Determination of folates in seaweeds by high-performance liquid chromatography.

    Science.gov (United States)

    Rodríguez-Bernaldo de Quirós, A; Castro de Ron, C; López-Hernández, J; Lage-Yusty, M A

    2004-04-02

    A reversed-phase high-performance liquid chromatography (HPLC) method, with a combination of fluorescence and ultraviolet detectors, to determine the folate forms present in commercial macroalgae products form northwest Spain as part of nutritional studies in dehydrated and canned seaweeds is reported. The method includes extraction of folates from seaweed by heat treatment, deconjugation of folate polyglutamates by incubation with hog kidney conjugase and purification by solid-phase extraction (SPE) with strong anion-exchange (SAX) cartridges. Separation was achieved with a Tracer Extrasil ODS 5 microm 25 cm x 0.4 cm column using acetonitrile and potassium phosphate buffer (pH 2.21 as mobile phase. Good results were obtained with respect to repeatability (relative standard deviation (R.S.D.) or = 90.80%). The amount of folate (as folic acid) in the six species ranged from 61.4 to 161.6 microg per 100 g dry mass. In all the seaweeds studied (Himanthalia elongata, Laminaria ochroleuca, Palmaria spp., Undaria pinnatifida and Porphyra spp. and Saccorhiza polychides) the single most abundant form is 5-CH3-H4-folate, except Porphyra and Himanthalia.

  6. Simultaneous determination of thiamine and riboflavin in edible marine seaweeds by high-performance liquid chromatography.

    Science.gov (United States)

    Sánchez-Machado, D I; López-Cervantes, J; López-Hernández, J; Paseiro-Losada, P

    2004-03-01

    This study presents a high-performance liquid chromatography (HPLC) method for simultaneous determination of thiamine and riboflavin and the results of its application to a number of edible seaweeds that are sampled in dried form (Himanthalia elongata, Laminaria ochroleuca, Undaria pinnatifida, Palmaria sp., and Porphyra sp.) or as canned food (H. elongata and Saccorhiza polyschides). Samples are prepared by acid and enzymatic hydrolysis. Optimized conditions for reversed-phase HPLC with fluorescence detection are as follow: column, Kromasil 100 C18; column temperature, 35 degrees C; mobile phase, a 72:28 (v/v) mixture of 0.005 M ammonium acetate (pH 6.7)-methanol; and flow rate, 1.35 mL/min. With these conditions, recovery is 95.52% for thiamine and 90.08% for riboflavin, and the method precision (relative standard deviation) is 2.66% for thiamine and 2.21% for riboflavin. On a dry weight basis, thiamine contents range from 0.14 microg/g in dried H. elongata to 2.02 microg/g in dried Porphyra and riboflavin contents from 0.31 microg/g in canned H. elongata to 6.15 microg/g in dried Porphyra.

  7. Simultaneous determination of eleven characteristic lignans in Schisandra chinensis by high-performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Junyang Hu

    2013-01-01

    Full Text Available Background: Schisandra chinensis, one of the well-known traditional Chinese herbal medicines, is derived from the dry ripe fruits of Schisandra chinensis (Turcz. Baill. according to the 9th China Pharmacopeia. Lignans are the main components isolated from extracts of S. chinensis and their content varies depending on where S. chinensis was collected. We have established a qualitative and quantitative method based on the bioactive lignans for control of the quality of S. chinensis from different sources. Materials and Methods: To develop a high-performance liquid chromatography method, an Elite ODS C18 column (250 mm Χ 4.6 mm, 5μm at a column temperature of 30°C and flow rate of 1.0ml/min using acetonitrile (A and water (B as the mobile phase with a linear gradient and the peaks were monitored at 217 nm. Results: All calibration curves showed good linearity (r ≥ 0.9995 within test ranges. This method showed good repeatability for the quantification of these eleven components in S. chinensis with intra- and inter-day relative standard deviations less than 0.43% and 1.21%, respectively. In the recovery test, results of accuracy ranged from 99.51% to 101.31% with RSD values less than 2.68%, Conclusion: The validated method can be successfully applied to quantify the eleven investigated components in 22 samples of S. chinensis from different sources.

  8. Preparation and Characterization of a New Quercetin-bonded Stationary Phase for High Performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    李来生; 方奕珊; 陈红; 张杨

    2012-01-01

    A quercetin-bonded silica gel stationary phase (QUSP) containing natural flavonoid ligand was first prepared via γ-glycidoxypropyltrimethoxysilane (KH-560) as a coupling reagent for high-performance liquid chromatography. Its chemical structure was characterized by Fourier infrared spectroscopy, elemental analysis, thermal thermogravimetry and 13C cross polarization/magic angle spinning nuclear magnetic resonance (CP/MAS NMR). The chromatographic property of QUSP was systematically evaluated by using neutral, basic and acidic aromatic com- pounds as probes. In order to clarify its retention mechanism, a comparative study of QUSP with conventional oc- tadecylsilyl-bonded stationary phase (ODS) was also carried out under the same conditions. The results showed that the new quercetin-bonded phase exhibited an excellent reversed-phase chromatographic property with relatively weak hydrophobicity. However, it has an advantage over ODS in the fast separation of polar aromatic compounds because the quercetin ligand could provide various sites besides hydrophobicity, such as hydrogen bonding, dipole-dipole, n-n staking and charge transfer interactions. QUSP was performed in the baseline separations of ion- ized polar basic or acidic compounds, including pyridines, anilines, pyrimidines, purines and phenols with symmet- ric peak shape in common mobile phases without buffer salt within relatively short time. The natural ligands from herbs are readily available and contain a variety of active sites, which facilitate the exploration of industrial chromatographic separation materials for green products.

  9. 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.

  10. Determination of mitomycin C in human aqueous humor and serum by high-performance liquid chromatography.

    Science.gov (United States)

    Li, W Y; Seah, S K; Koda, R T

    1993-09-08

    Mitomycin C (MMC) is used in the treatment of disseminated adenocarcinoma of the stomach and pancreas and is used in ophthalmology as adjunctive therapy in trabeculectomy. Since only small volumes of aqueous humor are available for analysis, a sensitive method requiring limited sample preparation was developed. An internal standard, 4-aminoacetophenone, was added to aqueous humor specimens, and the solution was directly injected into the high-performance liquid chromatographic (HPLC) column. The use of a short 50-mm C18 reversed-phase column gave adequate resolution of peaks with improved sensitivity. The method was applicable for determination of MMC in serum, although solid-phase extraction for sample clean-up was required prior to injection into the HPLC column, and analytical columns of 150-250 mm were necessary for adequate resolution of peaks. The method has been validated and is linear from 6.25 to 50 ng/ml in aqueous humor and from 10 to 500 ng/ml in serum.

  11. QUANTITATIVE ANALYSIS OF RAW MATERIAL OF TETRACYCLINE AND ITS RELATED SUBSTANCES BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

    Directory of Open Access Journals (Sweden)

    NABILA PERVEEN

    2006-01-01

    Full Text Available An isocratic method for the analysis of tetracycline (TC by high performance liquid chromatography (HPLC using polystyrene-divinylbenzene copolymer packing material is described. The method allows the complete separation of TC, 4-epitetracycline, anhydrotetracycline (ATC, 4-epianhydrotetracycline (EATC. A fermentation impurity, 2-acetyl-2-decarboxamidetetracycline is also resolved from TC. The mobile phase combines tert.-butanol, water and phosphate buffer, tetrabutylammonium sulphate and sodium ethylenediaminetetraacetate at pH 9.0 for elution at a temperature of 60°C. Among the organic modifiers examined, only tetrahydrofuran and tert.-butanol gave good results. For practical reasons, tert.-butanol was retained as the final organic modifier. The preliminary experiments were carried out on polystyrene-divinylbenzene copolymer packing material (PRP-I and PLRP-S columns. The selectivity of the columns is comparable. PLRP-S column was chosen as it shows better separation of tetracycline – anhydrotetracycline. The method was used to analyse the commercial samples.

  12. Determination of organic mercury species in soils by high-performance liquid chromatography with ultraviolet detection.

    Science.gov (United States)

    Hempel, M; Hintelmann, H; Wilken, R D

    1992-03-01

    Reversed-phase high-performance liquid chromatography with ultraviolet detection was optimized for the simultaneous separation and quantification of nine organic mercury compounds: methyl-, ethyl-, phenyl-, methoxyethyl-, ethoxyethyl-, benzoic and tolylmercury, mersalylic acid and nitromersol. The nine compounds were successfully separated on octadecylsilane columns (200 x 3 mm i.d.) by gradient elution with a methanol-water mixture ranging from 30 to 50% v/v. The detection limits for the various compounds are in the range 7.0-95.1 micrograms dm-3. For the extraction of five organomercurials from spiked soils, eight different extraction solutions were tested to differentiate between the total content and the available/soluble fraction of the analytes. Ammonium acetate solutions (1 mol dm-3) and water proved to be suitable agents for the estimation of the available and soluble fractions of methyl-, ethyl-, benzoic, methoxyethyl- and ethoxyethylmercury. For the determination of the total content of methyl- and benzoic mercury in soils, solutions of potassium iodide (1 mol dm-3)-ascorbic acid (0.1 mol dm-3) and oxalic acid (1 mol dm-3) provided recoveries in the ranges 53-81%. None of the solutions tested is suitable for the extraction of ethyl-, methoxyethyl- and ethoxyethylmercury.

  13. [Determination of clavulanic acid residue in milk by high performance liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Yang, Gang; Huang, Xianhui; Guo, Chunna; Fang, Qiuhua; He, Limin

    2012-06-01

    An analytical method was developed for the determination of clavulanic acid (CLAV) in milk by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). A 2 g milk sample was deproteinized by ethanol. The supernatant was transferred into a pear-shaped bottle to be evaporated to about 0.5 mL, and the residue was dissolved with ammonium acetate solution. The sample was determined by HPLC-MS/MS after the purification. The chromatographic separation was achieved on a Luna 5u C8 column using 0.1% formic acid in water and acetonitrile as mobile phases with gradient elution. The identification of CLAV was carried out by MS/MS equipped with electrospray ionization in negative scanning and multiple reaction monitoring (MRM) modes. Matrix-matched calibration standard was used for the quantification. The calibration curve showed perfect linear in the range of 10 - 400 microg/kg with the correlation coefficient of 0.999. The limit of detection (LOD, S/N > or = 3) was 10 microg/kg in milk, and the limit of quantification (LOQ, S/N > or = 10) was 20 microg/kg. The mean recoveries varied from 80.00% to 91.25% at the four spiked levels of LOQ, 1/2MRL (the maximum residue limit), MRL, and 2MRL with the relative standard deviations of 5.60% -8.77%. In conclusion, the established method can be applied for the determination of CLAV residues in milk.

  14. Biogenic amines in table olives. Analysis by high-performance liquid chromatography.

    Science.gov (United States)

    Hornero-Méndez, D; Garrido-Fernández, A

    1994-09-01

    Biogenic amines in fermented vegetables have scarcely been studied. Available data show that in table olives and fermented cucumbers their presence is rare and any determinations made have been restricted mainly to histamine. However, some microorganisms, especially those related to spoilage, found in the fermentation brines of such products may have amino acid decarboxylase activity and give rise to biogenic amines by unusual processes. A method for the simultaneous determination of eight biogenic amines (tryptamine, beta-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine, and spermine) has been developed to study their occurrence in fermented vegetables in more detail. The method consists of extraction of the amines from olive paste with 5% m/v trichloracetic acid and successive transfers into water-saturated n-BuOH and 0.1 mol l-1 HCl. An aliquot of this mixture is dried and derivatized with dansyl chloride. The dansyl derivatives are then analysed by high-performance liquid chromatography. Special emphasis has been given to optimization of the n-BuOH and 0.1 mol l-1 HCl extractions and to the derivatization conditions. By applying this method to the analysis of spoilt olives, the presence of some biogenic amines has been demonstrated. Thus a new method for monitoring the presence of biogenic amines during the fermentation of olives and for detecting anomalous fermentations is envisaged.

  15. Quality Evaluation of Potentilla fruticosa L. by High Performance Liquid Chromatography Fingerprinting Associated with Chemometric Methods.

    Directory of Open Access Journals (Sweden)

    Wei Liu

    Full Text Available 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.

  16. Quantitation of meloxicam in the plasma of koalas (Phascolarctos cinereus) by improved high performance liquid chromatography.

    Science.gov (United States)

    Kimble, Benjamin; Li, Kong Ming; Govendir, Merran

    2013-01-01

    An improved method to determine meloxicam (MEL) concentrations in koala plasma using reversed phase high performance liquid chromatography equipped with a photo diode array detector was developed and validated. A plasma sample clean-up step was carried out with hydrophilic- lipophilic copolymer solid phase extraction cartridges. MEL was separated from an endogenous interference using an isocratic mobile phase [acetonitrile and 50 mM potassium phosphate buffer (pH 2.15), 45 : 55 (v : v)] on a Nova-Pak C18 4-µm (300 × 3.9 mm) column. Retention times for MEL and piroxicam were 8.03 and 5.56 min, respectively. Peak area ratios of MEL to the internal standard (IS) were used for regression analysis of the calibration curve, which was linear from 10 to 1,000 ng/mL (r(2) > 0.9998). Average absolute recovery rates were 91% and 96% for MEL and the IS, respectively. This method had sufficient sensitivity (lower quantitation limit of 10 ng/mL), precision, accuracy, and selectivity for routine analysis of MEL in koala plasma using 250-µL sample volumes. Our technique clearly resolved the MEL peak from the complex koala plasma matrix and accurately measured MEL concentrations in small plasma volumes.

  17. 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.

  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. Determination of clomazone residues in soybean and soil by high performance liquid chromatography with DAD detection.

    Science.gov (United States)

    Hu, Jiye; Cao, Di; Deng, Zhubo

    2011-04-01

    A simple analysis method to detect clomazone residues in soybean and soil was developed using solid phase extraction coupled with high performance liquid chromatography with diode-array detection. The pesticide residues present in soybean and soil matrices were extracted with methanol-water and extracts purified with Florisil cartridges. The analytes from soybean and soil matrix were eluted with petroleum ether-acetic ether (10 mL, 95:5, v/v) and petroleum ether-acetic ether (2 mL, 95:5, v/v), respectively. The overall recovery of fortified soybean and soil at the levels of 0.01, 0.1 and 0.5 mg/kg ranged from 89.75% to 106.6%, and the coefficients of variation (CV) ranged from 1.68% to 4.93% (n = 3). The limit of quantification (LOQ) is 0.01 mg/kg. This method has been applied to the analysis of clomazone in real samples of soybean and soil. The dissipation of residue over the time in soil coincided with C = 1.189e(-0.0926t ) and the half-lives (T(1/2)) was 7.48 days. The final residue in soybean was lower than 0.01 mg/kg at harvest time. Direct confirmation of the analyte in real samples was achieved by gas chromatography-mass spectrometry.

  20. Detection of honey adulteration with starch syrup by high performance liquid chromatography.

    Science.gov (United States)

    Wang, Shaoqing; Guo, Qilei; Wang, Linlin; Lin, Li; Shi, Hailiang; Cao, Hong; Cao, Baosen

    2015-04-01

    According to saccharide profile comparison between starch syrups and pure honeys analysed through high performance liquid chromatography (HPLC), a characteristic peak was found at 15.25 min retention time in HPLC chromatogram of syrup, but no peak was observed at the same retention time in chromatogram of pure honeys. This characteristic peak for syrup was identified as an overlapping peak of oligosaccharides with more than 5 degree of polymerisation (DP) based on HPLC chromatogram comparison between starch syrup and a series of standard mono-, di- and oligosaccharides of 3-7 DP. Additionally syrup content correlated linearly with the height of the characteristic peak of syrup under different slope in two ranges 2.5-7.5% and 10-100%, respectively. Therefore, the characteristic peak at 15.25 min retention time can serve as a syrup indicator in HPLC analysis of the adulterated honeys. This new HPLC method for honey adulteration detection was further applied in an authenticity inspection on more than 100 commercial honeys. In addition to the improved accuracy of honey adulteration detection, the proposed HPLC method was simple, low cost and easy practice for honey product quality control by government department considering the popularity of HPLC device and technology. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. High-performance liquid chromatography of unmodified rosin and its applications in contact dermatology.

    Science.gov (United States)

    Sadhra, S; Gray, C N; Foulds, I S

    1997-10-24

    Rosin is a well recognised skin sensitiser and is also amongst the most common causes of occupational asthma. Due to its complex chemical composition, it is difficult to isolate its many components and this has hindered progress in the identification of the specific respiratory and contact allergens it contains. This paper reports the application of high-performance liquid chromatography and other analytical techniques to the isolation and identification of contact allergens in complex mixtures such as rosin. HPLC methods were developed in order to isolate as many rosin components as possible and these were then patch tested on rosin sensitive individuals. The structure of the most dermatologically active component was then determined using mass spectrometry, nuclear magnetic resonance and infrared techniques. An HPLC method has also been developed which will enable the identification of rosin in commercial products, providing a valuable tool for determining the cause of rosin contact allergy. Furthermore, mass spectral data for the common abieitic-type resin acids are compiled which were used to confirm the identification of the HPLC resin acid peaks and have not been reported previously.

  2. Determination of meropenem levels in human serum by high-performance liquid chromatography with ultraviolet detection.

    Science.gov (United States)

    Roth, Thomas; Fiedler, Stella; Mihai, Sidonia; Parsch, Hans

    2017-05-01

    Meropenem is a β-lactam broad-spectrum antibiotic and belongs to the subgroup of carbapenems. It is primarily used in intensive care units for intravenous treatment of severe infections. To avoid bacterial resistance or toxic side effects, the determination of serum meropenem concentration is highly advisable. A simple and fast method for the quantitative determination of meropenem in human serum using high-performance liquid chromatography with ultraviolet detection (HPLC/UV) was developed and validated. Meropenem was determined by an isocratic HPLC using a tris(hydroxymethyl)aminomethane buffer (pH 8.5; 15% methanol) as a mobile phase and UV detection at 300 nm, with a flow rate of 1.0 mL/min and an analysis time of 10 min. Chromatographic separation was performed on a Kinetex C18 column (5 μm, 150 × 4.6 mm). In order to remove undesired serum components, solid-phase extraction was used for sample preparation. Since meropenem is not stable in solution, sample and stock solution were stored at -80°C. After preparation, samples were stable at room temperature for at least 6 h. The calibration curve was linear from 3.5 to 200 mg/L with a correlation coefficient r(2) of 0.999. The method is accurate with an intra- and inter-assay precision <18.5%. Copyright © 2016 John Wiley & Sons, Ltd.

  3. Determination by high-performance liquid chromatography of phenylbutazone in samples of plasma from fighting bulls.

    Science.gov (United States)

    Marti, M I González; Sánchez, González C I; Jiménez, Hernández; Cachán, García; Castro, de Cabo M J; Cuadrado, A L Garzón

    2002-03-25

    The purpose of this study was to investigate the possible presence of phenylbutazone in plasma samples from fighting bulls killed in 2nd and 3rd category bullrings in the province of Salamanca (Spain) in 1998, 1999 and 2000. For quantitative and qualitative determination, a high-performance liquid chromatograph was used, equipped with a photodiode-array detector and setting wavelengths at 240, 254 and 284 nm. The mobile phase optimized for the simultaneous detection of dexamethasone, betamethasone, flunixin and phenylbutazone, was 0.01 M acetic acid pH 3 in methanol (35:65 v/v) at a flow rate of 1 ml/min. Plasma samples were deproteinized with 400 microl of acetonitrile and 20 microl of the supernatant were injected directly into the chromatographic system equipped with a Lichrospher 60 RP select B column and guard column. For the quantitative analysis, standard calibration curves were made in a concentration range between 0.25 and 30 microg/ml, using betamethasone as internal standard. The retention time of phenylbutazone was 8.7 +/- 0.2 min and recovery was 83%. The detection and quantification limits were 0.016 and 0.029, respectively for A=240 nm. The study results show that 17 of the 74 samples analyzed in 1998, 18 of those from 1999 and 10 of those from 2000 were positive for phenylbutazone.

  4. 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.

  5. Optimised determination of clobazam in human plasma with extraction and high-performance liquid chromatography analysis.

    Science.gov (United States)

    Bolner, A; Tagliaro, F; Lomeo, A

    2001-01-05

    The analysis of clobazam by high-performance liquid chromatography and UV detection is described herein. After adding an internal standard, 600 microl of plasma were extracted under basic conditions onto disposable cartridges packed with celite. The organic extract was then evaporated to dryness and the residue reconstituted in 200 microl of mobile phase. A 20 microl aliquot was injected into chromatograph. The HPLC system was equipped with an Ultrasphere C8 analytical column coupled with an UV detector set at 235 nm. The mobile phase was an acetate buffer 20 mM, pH 5.5, containing acetonitrile and triethylamine 70:30:0.01 (v/v); the flow-rate was 1.8 ml/min. Using this method, clobazam can be detected with a sensitivity limit of 6 ng/ml and the RSD% intra- and inter-assay were lower than 5%. For its ruggedness and reliability, the proposed method is particularly suitable for therapeutic drug monitoring in epilepsy.

  6. High-performance liquid chromatographic assay of free norepinephrine, epinephrine, dopamine, vanillylmandelic acid and homovanillic acid.

    Science.gov (United States)

    Gerlo, E; Malfait, R

    1985-09-13

    A procedure is described for the concurrent assay of free norepinephrine, epinephrine, dopamine, vanillylmandelic acid and homovanillic acid in physiological fluids using high-performance liquid chromatography with electrochemical detection. The column packing is an octadecyl-bonded silica. A single mobile phase containing 1-octanesulphonate is used for the assay of catecholamines and for the assay of the acidic metabolites. An efficient sample preparation scheme is presented for the isolation of the catecholamines and their acidic metabolites from the same sample aliquot. Catecholamines are extracted by ion exchange on small columns and adsorption on alumina, using dihydroxybenzylamine as an internal standard. Vanillylmandelic acid and homovanillic acid are recovered from the combined loading and washing effluents of the ion-exchange column by a solvent extraction procedure. Recovery of catecholamines averages 67%. The limit of detection for individual catecholamines is ca. 30 pg. Recoveries of vanillylmandelic acid and homovanillic acid average 77% and 87%, respectively. The use of the same mobile phase for the concurrent assay of catecholamines and their acidic metabolites considerably increases the throughput of samples in the chromatographic system by eliminating the time-consuming column-equilibration periods.

  7. Simultaneous determination of 11 preservatives in cosmetics by high-performance liquid chromatography.

    Science.gov (United States)

    Aoyama, Airin; Doi, Takahiro; Tagami, Takaomi; Kajimura, Keiji

    2014-10-01

    Preservatives prevent the growth of microorganisms in foods, pharmaceuticals and cosmetics. There exist numerous restrictions regarding the maximum allowable levels of preservatives in cosmetics. We analyzed 11 regulated preservatives in commercial cosmetics and manufacturers need to analyze their products for quality control purposes. However, methods used in previous studies to date have been inadequate for use by public institutions and manufacturers. Therefore, an effective, scalable method for the analysis of preservatives in cosmetics is required. We developed a novel method for the simultaneous determination of 11 regulated preservatives in cosmetics by high-performance liquid chromatography (HPLC). We applied the samples to a C18 column in a simple mobile phase (5 mmol/L ammonium formate solution and acetonitrile) with gradient elution at a flow rate of 1.0 mL/min at a single wavelength (230 nm). The correlation coefficients of the calibration curves were >0.997. The percent recoveries were 92.8-111.9% and the relative standard deviations were preservatives was >1.9. Because of the simple conditions for isolation and complete separation, the HPLC method can be effectively applied to the analysis of preservatives in commercially retailed cosmetics.

  8. High-Performance Liquid Chromatographic Ultraviolet Determination of Memantine Hydrochloride after In Vitro Transdermal Diffusion Studies

    Directory of Open Access Journals (Sweden)

    Sergio del Rio-Sancho

    2013-01-01

    Full Text Available The purpose of the present work was to validate an accurate and precise high-performance liquid chromatography (HPLC method involving ultraviolet detection for the quantitative analysis of memantine hydrochloride. In order to analyze a molecule with no chromophoric groups that could be detected by a UV/visible detector, it was necessary to extract the drug and to perform a dansylation reaction that enabled the UV/visible detection of the derivatized molecule. Separation was carried out with a 150 mm Kromasil C18 column at room temperature. The detection response, at 218 nm, was found to be linear in the concentration range from 0.5 to 50 μg/mL. The method was validated for specificity, linearity, precision, accuracy, limit of detection, limit of quantification, and robustness. The limit of detection (LOD was 0.144 μg/mL, and the limit of quantification (LOQ was 0.437 μg/mL. The dansylated memantine complex was stable for at least five days in all the conditions evaluated. The potential use of this method has been demonstrated by the quantification of memantine hydrochloride contained in samples from the study of its in vitro transdermal permeation.

  9. Rapid determination of propranolol enantiomers in rat plasma by column-switching-high performance liquid chromatography%柱切换技术-高效液相色谱法快速检测大鼠血浆中的普萘洛尔对映体

    Institute of Scientific and Technical Information of China (English)

    武晓玉; 王荣; 谢华; 王剑锋; 贾正平; 张强; 王先华

    2011-01-01

    A high performance liquid chromatographic ( HPLC) method with column-switching was developed and validated for rapid determination of two propranolol enantiomers in rat plasma. The column of restricted-access media was used as a pre-treatment column and a Chiralcel OD-RH was used as analytical column. The plasma samples were injected directly into the pre-treatment column to remove plasma protein and endogenous constituents as well as to retent the propranolol enantiomers in the column using the mobile phase of borate buffer (pH 8.5)-methanol (95: 5 , v/v) at the flow rate of 1. 0 mL/min. Then the propranolol enantiomers were transferred to the Chiralcel OD-RH column using the mobile phase of isopropanol-ethanol-O. 2 mmol/L borate buffer (pH 8. 5) (30: 30: 40, v/v/v) at a flow rate of 0. 8 mL/min by column-switching technology. The column-switching time was 3.0 min, the used wavelength was 293 nm and the column temperature was set at 25 ℃. The calibration curve showed excellent linear relationship (r=0.999 5) in the concentration range from 25 mg/L to 500 mg/L for propranolol enantiomers in plasma. The intra-day and inter-day assay precisions and accuracies were well and the relative standard deviations ( RSDs) were less than 5%. The average recoveries (n =6) of the two enantiomers at 3 spiked levels were from 97. 89% to 101. 56%. All the values of the method validation were within the generally accepted criteria for biological sample analysis.The results show that the method is convenient, quick, sensitive and accurate. The method was successfully applied in the determination of propranolol enantiomers in rat blood pharma-cokinetics study.%建立了快速检测大鼠血浆中普萘洛尔对映体浓度的柱切换-高效液相色谱法.将自制限进填料柱作为预处理柱,通过直接进样方式,使普萘洛尔对映体在预处理柱上保留,同时除去血浆中的蛋白质等大分子;再通过柱切换技术,使普萘洛尔对

  10. Preparation and characterization of high performance Schiff-base liquid crystal diepoxide polymer

    Energy Technology Data Exchange (ETDEWEB)

    Liu Huan; Fu Zien [Key Laboratory of Organic Polymer Material for Electronics, Guangzhou Institute of Chemistry, Chinese Academy of Sciences, P.O. BOX 1122, Guangzhou 510650 (China); Graduate School of the Chinese Academy of Sciences, Beijing 100049 (China); Xu Kai, E-mail: xk@gic.ac.cn [Key Laboratory of Organic Polymer Material for Electronics, Guangzhou Institute of Chemistry, Chinese Academy of Sciences, P.O. BOX 1122, Guangzhou 510650 (China); Cai Hualun; Liu Xin [Key Laboratory of Organic Polymer Material for Electronics, Guangzhou Institute of Chemistry, Chinese Academy of Sciences, P.O. BOX 1122, Guangzhou 510650 (China); Graduate School of the Chinese Academy of Sciences, Beijing 100049 (China); Chen Mingcai [Key Laboratory of Organic Polymer Material for Electronics, Guangzhou Institute of Chemistry, Chinese Academy of Sciences, P.O. BOX 1122, Guangzhou 510650 (China)

    2012-02-15

    Graphical abstract: The specific effects of highly conjugated Schiff-base moiety on thermal properties of the Schiff-base epoxy polymer were proposed first by us. From the point of view of structure-properties relationship, it can be considered that owing to the presence of the Schiff-base group, the high performance liquid crystal diepoxide polymer displayed improved thermal stability. Highlights: Black-Right-Pointing-Pointer In this work, we first proposed that specific effects of highly conjugated Schiff-base moiety on thermal properties of the Schiff-base epoxy polymer. Black-Right-Pointing-Pointer As one aim of this study, the thermal and thermal-oxidative stabilities of the thermosets were studied by TGA under nitrogen and under air. Black-Right-Pointing-Pointer The second aim of this study was to further understand the thermal degradation mechanism. Black-Right-Pointing-Pointer For thermal degradation mechanism of this polymer under nitrogen, TG-IR was used to investigate volatile components, and SEM/EDS was used to explore morphologies and chemical components of the residual char. Black-Right-Pointing-Pointer From the point of view of structure-properties relationship, it can be considered that owing to the presence of the Schiff-base group, the high performance liquid crystal diepoxide polymer displayed the improved thermal stability. - Abstract: A novel Schiff-base liquid crystal diepoxide polymer was prepared via a thermal copolymerization of a Schiff-base epoxy monomer (PBMBA) with a diamine co-monomer (MDA). We first proposed that specific effects of highly conjugated Schiff-base moiety on thermal properties of the Schiff-base epoxy polymer (PBMBA/MDA). Thermal degradation behavior of the polymer was characterized using thermogravimetric analysis (TGA) under nitrogen and under air, respectively. Thermogravimetric data obtained from TGA under nitrogen and under air reveal that PBMBA/MDA exhibits higher thermal stability compared with bisphenol-A type

  11. 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.

  12. Preparative isolation and analysis of alcohol dehydrogenase inhibitors from Glycyrrhiza uralensis root using ultrafiltration combined with high-performance liquid chromatography and high-speed countercurrent chromatography.

    Science.gov (United States)

    Chen, Miao; Liu, Liangliang; Chen, Xiaoqing

    2014-07-01

    A simple, rapid, and effective assay based on ultrafiltration combined with high-performance liquid chromatography and high-speed countercurrent chromatography was developed for screening and purifying alcohol dehydrogenase inhibitors from Glycyrrhiza uralensis root extract. Experiments were carried out to optimize binding conditions including alcohol dehydrogenase concentration, incubation time, temperature, and pH. By comparing the chromatograms, three compounds were found possessing alcohol dehydrogenase binding activity in Glycyrrhiza uralensis root. Under the target-guidance of ultrafiltration combined with the high-performance liquid chromatography experiment, liquiritin (1), isoliquiritin (2), and liquiritigenin (3) were separated by high-speed countercurrent chromatography using ethyl acetate/methanol/water (5:1:4) as the solvent system. The alcohol dehydrogenase inhibitory activities of these three isolated compounds were assessed; compound 2 showed strongest inhibitory activity with an IC50 of 8.95 μM. The results of the present study indicated that the combinative method using ultrafiltration, high-performance liquid chromatography and high-speed countercurrent chromatography could be widely applied for the rapid screening and isolation of enzyme inhibitors from complex mixtures.

  13. Quality assessment of Cinnamomi Ramulus by the simultaneous analysis of multiple active components using high-performance thin-layer chromatography and high-performance liquid chromatography.

    Science.gov (United States)

    Wu, Xiaoxue; He, Jiao; Xu, Huarong; Bi, Kaishun; Li, Qing

    2014-09-01

    A novel and improved method for the quality assessment of Cinnamomi Ramulus was developed and completely validated. The method was established using fingerprint technology and simultaneous quantitative determination of six main marker compounds including coumarin, cinnamic alcohol, cinnamic acid, 2-methoxy cinnamic acid, cinnamaldehyde, and 2-methoxy cinnamaldehyde in the herbal medicine for the first time. A newly developed high-performance thin-layer chromatography method, which achieved simultaneous definition of five marker components by comparing the colors and retardation factor values of the bands in high-performance thin-layer chromatography, was first used for the authentication of Cinnamomi Ramulus. The fingerprints of 26 batches of herbal samples from different regions of China showed very similar chromatographic patterns that were evaluated by similarity analysis and hierarchical clustering analysis. In addition, six marker compounds were simultaneously determined using single standard to determine multiple components by the relative response factors. Compared with the external standard method, the new quantitative method was validated to determine multiple compounds in 26 batches of Cinnamomi Ramulus samples. All results demonstrated that the simple and rapid method could be effectively utilized for the quality control of Cinnamomi Ramulus. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Development of the fingerprints for the quality evaluation of Viscum coloratum by high performance liquid chromatograp%Development of the fingerprints for the quality evaluation of Viscum coloratum by high performance liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    Yun-Li Zhao; Rong-Hua Fan; Hong-Xia Yuan; Miao Yu; Kai-Shun Bi; Zhi-Guo Yu

    2011-01-01

    A high-performance liquid chromatography coupled ultraviolet (HPLC-UV) method was developed for a chemical fingerprint analysis of Viscum coloratura. Eighteen peaks were selected as the common peaks and Homoeriodictyol-7-O-β-D-apiosiyl-(1→2)-β-D-glucoside

  15. Speciation of arsenic using chelation solvent extraction and high performance liquid chromatography

    Energy Technology Data Exchange (ETDEWEB)

    Cathum, Shamil J.; Obenauf, Alison; Punt, Monique [SAIC Canada, Ottawa, Ontario (Canada); Brown, Carl E. [Emergencies Engineering Technology Office, Environmental Technology Centre, Environment Canada, Ottawa, Ontario (Canada)

    2007-02-15

    Research interest in speciation of arsenic stems from its species dependent behavior in the environment and in living organisms. The complexity of the matrix to be analyzed and low concentrations of target arsenic species that may be labile or difficult to chromatogram, indicate that a suitable pre-treatment methodology is required. This study investigated the usefulness of chelation solvent extraction - high performance liquid chromatography (CSE-HPLC) for the speciation of arsenic in water. It involved reacting arsenic with the chelant known for its affinity towards arsenic, followed by extraction, separation, and identification of the arsenic-chelant-arsenic complex. Arsenic species having different physicochemical properties were investigated. Species, such as, As{sub 2}O{sub 3}, As{sub 3}O{sub 5}, KH{sub 2}AsO{sub 4}, Na{sub 2}HAsO{sub 4}, and NaAsO{sub 2}were detected as a group of closely eluted peaks with different retention times and spectral properties, whereas, the organic arsenic species CH {sub 3}Na {sub 2}AsO {sub 3}, o-arsanilic acid, roxarson and triphenyl arsine separated quite well on the EnviroseP-CM HPLC column. Key method parameters, such as, type of HPLC column, composition of mobile phase and organic solvents affecting peak resolution and sensitivity were optimized. Real environmental matrices contaminated with arsenic were analyzed under varying wavelengths ({lambda}{sub max} = 190, 210, 220, 234, 244, and 282 nm), with good precision. Different arsenic species were detected in these samples with excellent background and signal-to-noise ratios demonstrating the robustness of the method. The detection limit, reproducibility, selectivity, accuracy, and dynamic range of the calibration curves were evaluated. (Abstract Copyright [2007], Wiley Periodicals, Inc.)

  16. Pharmacokinetic study of mangiferin in rat plasma and retina using high-performance liquid chromatography

    Science.gov (United States)

    Hou, Yunlong; Fan, Shengjun; Gu, Yuanqin; Yu, Xuhui; Li, Baoxin

    2010-01-01

    Purpose Although the naturally occurring antioxidant mangiferin has been widely used, it is not yet known whether it can cross the blood-retina barrier (BRB) and enter the eye. The purpose of this experiment was to investigate the ability of mangiferin to pass the blood-retina barrier. Methods Sprague–Dawley rats were used for biologic fluid sampling after intravenous administration of mangiferin at doses of 10, 25, and 50 mg/kg. Blood and retina samples were collected at different time points post-dose. High-performance liquid chromatography (HPLC) separation was conducted on a COSMOSIL 5C18—MS—II column (4.6 mm×250 mm, 5 μm) with a flow rate of 1.0 ml/min using a mobile phase comprised of methanol −2% glacial acetic acid (40:60 v:v). Results The HPLC method has proven suitable to determine the presence of mangiferin in the eye. The plasma concentration of mangiferin was dose dependent. Pharmacokinetic parameters of mangiferin in plasma after intravenous administration were fitted to the two-compartment model with the first-order elimination and first-order transfer between central and peripheral compartments. The concentration of mangiferin in the retina goes with that in the blood. Mangiferin concentrations in the retina reached 5.69±1.48 μg/ml 0.5 h after intravenous administration (50 mg/kg) and then dropped gradually to 0.30±0.02 μg/ml 5.0 h later. The eye–to-plasma concentration ratio was 2.80%. Conclusions Mangiferin can pass the blood-retina barrier after a single intravenous administration and may be a potential natural antioxidant in treating eye diseases. PMID:20806037

  17. DNA-based diagnosis of isolated sulfite oxidase deficiency by denaturing high-performance liquid chromatography.

    Science.gov (United States)

    Lam, Ching-Wan; Li, Chi-Keung; Lai, Chi-Kong; Tong, Sui-Fan; Chan, Kwok-Yin; Ng, Grace Sui-Fun; Yuen, Yuet-Ping; Cheng, Anna Wai-Fun; Chan, Yan-Wo

    2002-01-01

    Isolated sulfite oxidase deficiency is a rare autosomal recessive disease, characterized by severe neurological abnormalities, seizures, mental retardation, and dislocation of the ocular lenses, that often leads to death in infancy. There is a special demand for prenatal diagnosis, since no effective treatment is available for isolated sulfite oxidase deficiency. Until now, the cDNA sequence of the sulfite oxidase (SUOX) gene has been available, but the genomic sequence of the SUOX gene has not been published. In this study, we have performed a DNA-based diagnosis of isolated sulfite oxidase deficiency in a Chinese patient. To do so, we designed oligonucleotide primers for amplification of the predicted exons and intron-exon boundaries of the SUOX gene obtained from the completed draft version of the human genome. Using overlapping PCR products, we confirmed the flanking intronic sequences of the coding exons and that the entire 466-residue mature peptide is encoded by the last exon of the gene. We then performed mutation detection using denaturing high-performance liquid chromatography (DHPLC). The DHPLC chromatogram of exon 2b showed the presence of heteroduplex peaks only after mixing of the mutant DNA with the wild-type DNA, indicating the presence of a homozygous mutation. Direct DNA sequencing showed a homozygous base substitution at codon 160, changing the codon from CGG to CAG, which changes the amino acid from arginine to glutamine, i.e., R160Q. The DNA-based diagnosis of isolated sulfite oxidase deficiency will enable us to make an accurate determination of carrier status and to perform prenatal diagnosis of this disease. The availability of the genomic sequences of human genes from the completed draft human genome sequence will simplify the development of molecular genetic diagnoses of human diseases from peripheral blood DNA.

  18. Analysis of Some Pesticide Residues in Cauliflower by High Performance Liquid Chromatography

    Directory of Open Access Journals (Sweden)

    Sheheli Islam

    2009-01-01

    Full Text Available Problem statement: Increased use of chemicals on vegetables started gaining momentum and continued its up-trend in Bangladesh. Wide spread use of pesticides in agriculture concern of residue accumulation, which may remain in food and agricultural environment causing concern of human health and risking ecological balance. Attempt made to ensure that their applications were correct and safe and result in no residues in food beyond codex developed maximum residue limits. Approach: This study reported a method based on High Performance Liquid Chromatography (HPLC for determination of pesticide residues used in Cauliflower. Cauliflower sprayed with, 4 different pesticides (diazinon, malathion, chlorpyrifos and cypermethrin at recommended dose and double of recommended dose were analyzed for their residual contents. Samples were collected at same day after application of pesticide. Commercial samples of cauliflowers were collected from different markets of Dhaka city. Reversed-phase HPLC system with UV detection was used for the separation, identification and quantification of all these analytes using acetonitrile-water (70:30, v/v as mobile phase. Results: Limit of detection of 0.02 mg kg-1 was obtained. Calibration curves that constructed for the analytes spiked into samples followed linear relationships with good correlation coefficients (R2>0.990. In the analysis, from vegetables treated with diazinon and chlorpyrifos at recommended and double of recommended doses, residual amounts above respective MRL values were found. Conclusion: Method used permitted the determination of these pesticides in cauliflower at concentration level demanded by current legislation. Attention paid on excess use or abuse of pesticides by judicious application for safety of public health in Bangladesh. Additional data to monitor residues in food and to fill gaps in current knowledge would be helpful in assessing human exposure risks from ingestion of contaminated

  19. Simultaneous determination of cyclodol and diprazin by thin layer chromatography and high performance liquid chromatography.

    Science.gov (United States)

    Makharadze, R; Adeishvili, L; Chelidze, T; Imnadze, N; Nizharadze, N

    2009-11-01

    Ciklodol (trihexyphenidil)--the central and peripheral m-cholinoblocker is currently used with other antipsychotic drugs such as phenotiazines and tricycle antidepressants. For the purpose of simultaneous determination of ciklodol and diprazine, were selected two methods of analysis: Thin Layer Chromatography (TLC) and High Performance Liquid Chromatography (HPLC). During development of TLC method was studied the 10 visualizing system and 24 mobile systems. For individual or simultaneous determination of ciklodol and diprazine were recommended the following solvents' systems: 1. Toluene-acetone-ethanole-25%NH(4)OH (45:45: 7.5:2.5), 2. Hexane-ethyl acetate (15:5), 3. Chloroform-heptene-25%NH(4)OH (16:3:3), 4. Ethylacetate-hexane (10:10), 5. Acetonitrile-metanol (10:10) and 6.Heptene-chloroform-ethanol-25% NH(4)OH (5:10:3:1). As visualizing systems were chosen: Iodine vapors, blacklight (UV254) and reagent of FNP. Reagent of FNP gives colored spot just with diprazine and it is also could be used for separation of both objects in simultaneous analysis. Developed HPLC method of simultaneous determination of ciklodol and diprazine: like mobile phase is recommended: Acetonitril- 0.05M KH(2)PO4 (55:45) (v/v) +H(3)PO(4) (pH3.5), column EC250 x 4.6mm, with solid phase Nucleosil, flow rate 1ml/min, sample volume 40 microl. In given conditions, the retention time of ciklodol is 6.005min and diprazine 7.227min. Developed method of simultaneous determination and separation of ciklodol and diprazine in respective mixtures could be successfully applied as in the pharmaceutical, as well in the chemical-toxicological laboratories.

  20. Ambient temperature nanoelectrospray ion mobility detector for high performance liquid chromatography in determining amines.

    Science.gov (United States)

    Chen, Chuang; Hou, Keyong; Wang, Weiguo; Li, Jinghua; Li, Haiyang

    2014-09-01

    A nanoelectrospray ionization ion mobility spectrometer (nanoESI-IMS) working at ambient pressure and ambient temperature was developed as a detector of high performance liquid chromatography (HPLC) to achieve sensitive detection of amines with no derivatization and meanwhile provide another dimension of separation. The easier desolvation property of the charged droplets formed in nanoESI source enabled complete desolvation of the product ions of sixteen amines and drugs using the nanoESI-IMS at ambient temperature. Working at ambient temperature was good for suppressing the dissociation of thermal volatile ions, such as only the proton adducted molecular ions were observed for morphine in the nanoESI-IMS. Besides, the resolving power of the nanoESI-IMS also showed an increasing tendency as lowering the working temperature, an increment of 19 percent and 10 percent was observed for diethylamine and triethylamine as the temperature dropped from 92°C to 32°C. The resolving power of the nanoESI-IMS at 32°C for the 16 tested compounds was amid 33-44. With the nanoESI-IMS coupled to HPLC, a six-compound mixture including isomers was successfully separated and detected without any derivatization. And linear response ranges of 1 to 20, 0.5 to 20, and 0.8 to 20μgml(-1) and limits of detection of 0.25, 0.15, and 0.17μgml(-1) for triethylamine, diethylamine, and butylamine, respectively, were obtained with the hyphenated system. These results showed the excellent performance of the two-dimensional separation and detection method in direct qualitative and quantitative analyses of amines.

  1. Advanced use of high-performance liquid chromatography for synthesis of controlled metal clusters.

    Science.gov (United States)

    Niihori, Yoshiki; Matsuzaki, Miku; Uchida, Chihiro; Negishi, Yuichi

    2014-07-21

    Because the synthesis of metal clusters with multiple ligand types results in a distribution of ligands, high-resolution separation of each unique cluster from the mixture is required for precise control of the ligand composition. Reverse-phase high-performance liquid chromatography combined with appropriate transitioning of the mobile phase composition is an extremely effective means of separating ligand combinations when working with metal clusters protected by two different types of thiolates. We report herein advanced use of this method. The studies involving Au₂₄Pd(SR₁)₁₈-x(SR₂)x and Au₂₄Pd(SR₁)₁₈-x(SeR₂)x (SR₁, SR₂ = thiolate, SeR₂ = selenolate) revealed the following. (1) In general, an increase in the difference between the polarities of the functional groups incorporated in the two types of ligands improves the separation resolution. A suitable ligand combination for separation can be predicted from the retention times of Au₂₄Pd(SR₁)₁₈ and Au₂₄Pd(SR₂)₁₈, which cause the terminal peaks in a series of peaks. (2) The use of a step-gradient program during the mobile phase substitution results in improved resolution compared to that achievable with the linear gradients applied in prior work. (3) This technique is also useful for the evaluation of the chemical compositions of metal clusters protected by two different types of ligands with similar molecular weights. These findings will provide clear design guidelines for the functionalization of metal clusters via control of the ligand composition, and will also improve our understanding of the high-resolution isolation of metal clusters.

  2. 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).

  3. 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.

  4. Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns.

    Science.gov (United States)

    Jones, Andrew; Pravadali-Cekic, Sercan; Hua, Stanley; Kocic, Danijela; Camenzuli, Michelle; Dennis, Gary; Shalliker, Andrew

    2016-04-26

    A protocol for the use of reaction flow high performance liquid chromatography columns for methods employing post column derivatization (PCD) is presented. A major difficulty in adapting PCD to modern HPLC systems and columns is the need for large volume reaction coils that enable reagent mixing and then the derivatization reaction to take place. This large post column dead volume leads to band broadening, which results in a loss of observed separation efficiency and indeed detection in sensitivity. In reaction flow post column derivatization (RF-PCD) the derivatization reagent(s) are pumped against the flow of mobile phase into either one or two of the outer ports of the reaction flow column where it is mixed with column effluent inside a frit housed within the column end fitting. This technique allows for more efficient mixing of the column effluent and derivatization reagent(s) meaning that the volume of the reaction loops can be minimized or even eliminated altogether. It has been found that RF-PCD methods perform better than conventional PCD methods in terms of observed separation efficiency and signal to noise ratio. A further advantage of RF-PCD techniques is the ability to monitor effluent coming from the central port in its underivatized state. RF-PCD has currently been trialed on a relatively small range of post column reactions, however, there is currently no reason to suggest that RF-PCD could not be adapted to any existing one or two component (as long as both reagents are added at the same time) post column derivatization reaction.

  5. Rayleigh light scattering detection of three α1-adrenoceptor antagonists coupled with high performance liquid chromatograph

    Science.gov (United States)

    Li, Ai Ping; Peng, Huanjun; Peng, Jing Dong; Zhou, Ming Qiong; Zhang, Jing

    2015-08-01

    Herein, a Rayleigh light-scattering (RLS) detection method combined with high performance liquid chromatograph (HPLC) without any post-column probe was developed for the separation and determination of three α1-adrenoceptor antagonists. The quantitative analysis is benefiting from RLS signal enhancement upon addition of methanol which induced molecular aggregation to form an hydrophobic interface between aggregates and water that produce a sort of superficial enhanced scattering effect. A good chromatographic separation among the compounds was achieved using a Gemini 5u C18 reversed phase column (250 mm × 4.6 mm; 4 μm) with a mobile phase consisting of methanol and ammonium acetate-formic acid buffer solution (25 mM; pH = 3.0) at the flow rate of 0.7 mL min-1. The RLS signal was monitored at λex = λem = 354 nm. A limit of detection (LOD) of 0.065-0.70 μg L-1 was reached and a linear range was found between peak height and concentration in the range of 0.75-15 μg L-1 for doxazosin mesylate (DOX), 0.075-3.0 μg L-1 for prazosin hydrochloride (PRH), and 0.25-5 μg L-1 for terazosin hydrochloride (TEH), with linear regression coefficients all above 0.999. Recoveries from spiked urine samples were 88.4-99.0% which is within acceptable limits. The proposed method is convenient, reliable and sensitive which has been used successfully in human urine samples.

  6. [Determination of aflatoxins in cashew by high performance liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Bi, Ruifeng; Fan, Zhixian; Fu, Meng

    2011-12-01

    A method for the determination of four aflatoxins in cashew using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed. The sample was extracted with methanol-water (8: 2, v/v) solution, followed by a cleanup procedure with Florisil column. The target compounds were eluted using 5 mL acetone-water-formic acid (96: 3.5:0.5, v/v/v) solution. The eluate was dried under N2, then dissolved in 1 mL methanol. Four aflatoxins were separated in MG C18 column (100 mm x 3.0 mm, 3 microm) adopting a gradient program within 15 min. A triple quadrupole mass spectrometry equipped with an electrospray ionization source operated in the positive ion mode was used to detect the aflatoxins. The good correlation coefficients (r2 > 0.997) of the four aflatoxins were obtained within their respective linear ranges. The limits of detection (S/N = 3) were between 0.009 microg/kg and 0.04 microg/kg, and the limits of quantification (S/N = 10) were between 0.03 microg/kg and 0.12 microg/kg. The recoveries were in a range of 63.0% -78.5% with the relative standard deviations (RSDs) varied from 2.8% to 9.1%. The validation results meet the requirements of trace assay. Matrix effects were estimated and the signal suppression/enhancement ranged from 88.8% to 99.4%. The results indicate that the developed method is simple, fast, accurate, and can be applied for the determination of fours aflatoxins in cashew.

  7. Chiral stationary phases based on chitosan bis(methylphenylcarbamate)-(isobutyrylamide) for high-performance liquid chromatography.

    Science.gov (United States)

    Tang, Sheng; Bin, Qin; Chen, Wei; Bai, Zheng-Wu; Huang, Shao-Hua

    2016-04-01

    A series of chitosan bis(methylphenylcarbamate)-(isobutyrylamide) derivatives were synthesized by carbamylating chitosan isobutyrylamide with different methylphenyl isocyanates. Then the prepared chitosan derivatives were coated onto 3-aminopropyl silica particles, resulting in a series of new chiral stationary phases (CSPs) for high-performance liquid chromatography. It was observed that the chiral recognition abilities of these coated-type CSPs depended very much on the substituents on the phenyl moieties of the chitosan derivatives, the eluent composition, as well as the structure of racemates. As a typical example, the eluent tolerance of the prepared CSP with the best enantioseparation ability was investigated in detail, and the results revealed that the CSP exhibited extraordinary solvent tolerance and could still work without significant loss in enantioseparation capability after being flushed with chloroform (100%), ethyl acetate (100%) and even THF/n-hexane (70/30, v/v), while the traditional coated-type CSPs based on the cellulose and amylose derivatives, such as cellulose tris(3,5-dimethylphenylcarbamate) (CDMPC) and amylose tris(3,5-dimethylphenylcarbamate) (ADMPC), might be dissolved or highly swollen in these eluents. Therefore, the application of the resultant CSPs could address the problem of the dissolution and high swelling of traditional coated-type CSPs in some unusual eluents, broadening the possibility of eluent choice. In addition, a comparison of the prepared CSPs with the well known CDMPC- and ADMPC- based CSPs concerning the chiral recognition ability was also made. Separation performances achieved on the as-prepared CSPs in different eluents were found to be even superior to CDMPC- and ADMPC-based CSPs for the tested chiral compounds. In summary, we could safely draw the conclusion that the CSPs derived from chitosan isobutyrylamide derivatives were capable of excellent chiral recognition ability, and meanwhile possessed satisfactory

  8. Enantiomeric Separations of Pyriproxyfen and its Six Chiral Metabolites by High-Performance Liquid Chromatography.

    Science.gov (United States)

    Zhang, Chuntao; Liu, Hui; Liu, Donghui; Wang, Liying; Gao, Jing; Zhou, Zhiqiang; Wang, Peng

    2016-03-01

    Pyriproxyfen is a chiral insecticide, and over 10 metabolites have been identified in the environment. In this work the separations of the enantiomers of pyriproxyfen and its six chiral metabolites were studied by high-performance liquid chromatography (HPLC). Both normal phase and reverse phase were applied using the chiral columns Chiralpak IA, Chiralpak IB, Chiralpak IC, Chiralcel OD, Chiralcel OD-RH, Chiralpak AY-H, Chiralpak AD-H, Chiracel OJ-H, (R,R)-Whelk-O 1, and Lux Cellulose-3. The effects of the chromatographic parameters such as mobile phase composition and temperature on the separations were investigated and the enantiomers were identified with an optical rotation detector. The enantiomers of these targets could obtain complete separations (resolution factor Rs > 1.5) on Chiralpak IA, Chiralpak IB, Chiralcel OD, Chiralpak AY-H, or Chiracel OJ-H under normal conditions. Chiralcel OJ-H showed the best chiral separation results with n-hexane as mobile phase and isopropanol (IPA) as modifier. The simultaneous enantiomeric separation of pyriproxyfen and four chiral metabolites was achieved on Chiralcel OJ-H under optimized condition: n-hexane/isopropanol = 80/20, 15°C, flow rate of 0.8 ml/min, and UV detection at 230 nm. The enantiomers of pyriproxyfen and the metabolites , , and obtained complete separations on Chiralpak IA, Chiralpak IC, and Lux Cellulose-3 under reverse phase using acetonitrile/water as the mobile phase. The retention factors (k) and selectivity factors (α) decreased with increasing temperature, and the separations were better under low temperature in most cases. The work is of significance for the investigation of the environmental behaviors of pyriproxyfen on an enantiomeric level.

  9. Pharmacokinetics of mitomycin C in dogs: application of a high-performance liquid chromatographic assay.

    Science.gov (United States)

    Barbhaiya, R H; Papp, E A; Van Harken, D R; Smyth, R D

    1984-09-01

    A normal-phase high-performance liquid chromatographic (HPLC) assay was developed for the determination of mitomycin C in plasma and urine. The method involves extraction of mitomycin C from plasma or urine into ethyl acetate-2-propanol-chloroform (70:15:15) with UV detection at 365 nm. Quantitation was performed with an internal standard (porfiromycin) by the peak height ratio method. Excellent correlation was obtained between the HPLC assay and the established microbiological cup-plate bioassay. The pharmacokinetics of mitomycin C were investigated in beagle dogs following a 1-mg/kg iv (22-mg/m2) bolus dose. The plasma mitomycin C concentration versus time data were analyzed by using an open three-compartment model. The average volume of distribution was 1.90 L or 17% of body weight for the central compartment and 7.7 L or 68% of body weight for the terminal elimination phase. The volumes of distribution at steady state, calculated by model-dependent and -independent methods, compared very well with each other and were 6.5 L or 58% of body weight. Total body clearance averaged 112 mL/min, and the mean terminal plasma half-life was 53 min. The 0-24-h urinary excretion of intact mitomycin C accounted for 19% of the dose. The terminal half-life and percent urinary recovery of mitomycin C in dogs is similar to that in humans. Based on these observations, the dog appears to be a good model for studying the disposition of mitomycin C.

  10. Determination of plasma lactic acid concentration and specific activity using high-performance liquid chromatography.

    Science.gov (United States)

    Bleiberg, B; Steinberg, J J; Katz, S D; Wexler, J; LeJemtel, T

    1991-08-23

    Assessment of lactate metabolism is of particular interest during exercise and in disease states such as diabetes, shock, and absorptive abnormalities of short-chain fatty acids by the colon. We describe an analytical method that introduces radio-active tracers and high-performance liquid chromatography (HPLC) to simultaneously analyze concentrations and specific activities (SAs) of plasma lactate. The HPLC conditions included separation on a reversed-phase column (octadecylsilane) and an isocratic buffer (30% acetonitrile in water). [3H]Acetate served as an internal standard. Lactate and acetate were extracted from plasma samples with diethyl ether following a pH adjustment to less than 1.0 and back-extracted into a hydrophilic phase with sodium carbonate (2 mM, pH greater than 10.0). Lactate is detected in the ultraviolet range (242 and 320 nm) by derivatization with alpha-bromoacetophenone. Control plasma samples were studied after an overnight fast for precision and analytical recovery. Calibration curves were linear in the range 0.18-6.0 mM (r = 0.92). The precision was 3% and the analytical recovery was 87%. The detection limit of the method was 36 pmol. Determination of lactate metabolism was performed in a patient with chronic congestive heart failure who was administered primed-continuous L-[U-14C]lactate (10 microCi bolus and 0.3 microCi/min continuously) during a 60-min rest period. Mean arterial lactate concentration and SA were 1.69 +/- 0.2 mM and 253.8 +/- 22 dpm/mumol, respectively. Systemic lactate turnover was 25.65 mumol/kg per min. Lactic acid systemic turnover, organ uptake and release rates can be accurately determined by isocratic HPLC.

  11. Simultaneous determination of multiple marker constituents in concentrated Gegen Tang granule by high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Yip Yuekeung

    2007-06-01

    Full Text Available Abstract Background Concentrated Gegen Tang (GT granule is a widely available traditional Chinese medicinal product for the treatment of cold and flu. There was no reliable analytical method available for the quality assessment of GT granules. Methods An HPLC method with an Agilent Zorbax SB-Phenyl Stablebond column (250 × 4.6 mm, 5 μm was developed and validated. The mobile phase gradient was a mixture of 0.1% trifluoroacetic acid (TFA in acetonitrile (ACN and 0.1% TFA in water. The detection with a diode-array detector was set at 207, 230, 250 and 275 nm. Seven components, namely puerarin, daidzein, paeoniflorin, cinnamic acid, glycyrrhizin, ephedrine and pseudoephedrine were selected as marker compounds for the evaluation. Results The regression equations revealed good linear relationships (correlation coefficients: 0.9994–0.9998 between the peak areas and concentrations. The recovery was between 98.8% and 101.7% with good precision and accuracy. The quality of GT granule from four different manufacturers was evaluated with this newly developed method. Samples from four manufacturers showed similar profiles but contents of the detected markers varied significantly among manufacturers and batches. Conclusion A new method using high performance liquid chromatography (HPLC has been developed for simple and reliable quality control of commercial concentrated GT granules. Sensitivity was increased by multi-wavelength detection. The contents of selected marker components in GT granules varied significantly among manufacturers and batches, making it necessary to evaluate the quality of concentrated GT granules in the market.

  12. Simultaneous determination of three sesquiterpene lactones in Aucklandia lappa Decne by high performance liquid chromatography

    Directory of Open Access Journals (Sweden)

    Chang-Seob Seo

    2015-01-01

    Full Text Available Background: Aucklandia lappa Decne, a well known traditional herbal medicine, is used for the treatment of asthma, rheumatism, coughs, tuberculosis, and many other diseases. Objective: We performed simultaneous analysis of three sesquiterpene lactones, costunolide (1, dehydrocostus lactone (2, and alantolactone (3, obtained from a 70% methanol extract of A. lappa using high performance liquid chromatography–photodiode array (HPLC–PDA techniques. Materials and Methods: The compounds 1–3 were separated using a reversed phase SunFire™ C18 analytical column kept at 35°C by the isocratic elution with distilled water and acetonitrile as mobile phase. The flow rate was 1.0 mL/min, and the injection volume was 10 μL. Results: The established analytical method showed high linearity, with a correlation coefficient ≥0.9999. The limit of detection and the limit of quantification of compounds 1–3 were 0.06–0.13 μg/mL and 0.21–0.42 μg/mL, respectively. The recovery of the compounds 1–3 was 97.27–103.00%. The intra and inter day relative standard deviations were 0.09–0.97% and 0.09–1.06%, respectively. The amounts of the compounds 1–3 were 17.32, 28.26, and 0.01 mg/g, respectively. Conclusion: The established and validated HPLC–PDA method may be help for the quality control of herbal medicine, A. lappa.

  13. 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.

  14. Ultra-high-performance liquid chromatography profiling method for chemical screening of proanthocyanidins in Czech hops.

    Science.gov (United States)

    Olšovská, J; Kameník, Z; Čejka, P; Jurková, M; Mikyška, A

    2013-11-15

    Hops represent an important natural source of bioactive polyphenols, particularly proanthocyanidins, which can contribute to prevention of several civilization diseases, owing to their antioxidant and radical scavenging activity. We have developed a high-throughput ultra-high-performance liquid chromatography time-of-flight mass spectrometry profiling method, which can be used for monitoring of bioactive proanthocyanidins in hops. The method was applied for analysis of hops of four Czech varieties (Saaz, Sladek, Preminat and Agnus) from the 2011 crop (9 localities, 11 samples) and the 2012 crop (24 localities, 40 samples). Hop samples were extracted by acetone and the analytes were separated on the Acquity UPLC BEH Shield RP18 column. Partial validation of the method revealed a satisfactory intra-day repeatability of the method for retention times (relative standard deviation within 1.39%) as well as areas under the peaks (within 9.89%). Experimental data were evaluated using principal component analysis and cluster analysis. Significant amounts of di-, tri- and tetramer proanthocyanidins consisting of (epi)catechin and (epi)gallocatechin were found in the hop samples. The dependence of the proantocyanidin composition on both the variety and the growing locality was observed. Specifically, the traditional Saaz variety contained more frequently oligomers formed by (epi)catechin units only, whereas the varieties Premiant and Agnus produced oligomers consisting of (epi)catechin as well as (epi)gallocatechin units. The relative abundance of proanthocyanidins in studied hop varieties from the two crops, 2011 and 2012, did correspond to each other. In the further perspective, the method may also be used for prediction of qualitative marks or authenticity verification of hops.

  15. Dynamic Behavior of Clobazam on High-Performance Liquid Chromatography Chiral Stationary Phases.

    Science.gov (United States)

    Sabia, Rocchina; De Martino, Michela; Cavazzini, Alberto; Villani, Claudio

    2016-01-01

    Clobazam, a 1,5-benzodiazepin-2,4-dione, is a chiral molecule because its ground state conformation features a nonplanar seven-membered ring lacking reflection symmetry elements. The two conformational enantiomers of clobazam interconvert at room temperature by a simple ring-flipping process. Variable temperature HPLC on the Pirkle type (R)-N-(3,5-dinitronenzoyl)phenylglycine and (R,R)-Whelk-O1 chiral stationary phases (CSPs) allowed us to separate for the first time the conformational enantiomers of clobazam and to observe peak coalescence-decoalescence phenomena due to concomitant separation and interconversion processes occurring on the same time scale. Clobazam showed temperature dependent dynamic high-performance liquid chromatography (HPLC) profiles with interconversion plateaus on the two CSPs indicative of on-column enantiomer interconversion. (enantiomerization) in the column temperature range between Tcol = 10°C and Tcol = 30°C, whereas on-column interconversion was absent at temperature close to or lower than Tcol = 5°C. Computer simulation of exchange-deformed HPLC profiles using a program based on the stochastic model yielded the apparent rate constants for the on-column enantiomerization and the corresponding free energy activation barriers. At Tcol = 20°C the averaged enantiomerization barriers, ΔG(‡), for clobazam were found in the range 21.08-21.53 kcal mol(-1) on the two CSPs. The experimental dynamic chromatograms and the corresponding interconversion barriers reported in this article are consistent with the literature data measured by DNMR at higher temperatures and in different solvents.

  16. Validation of stability indicating high performance liquid chromatographic method for estimation of Desloratadine in tablet formulation

    Directory of Open Access Journals (Sweden)

    R.B. Patel

    2017-02-01

    Full Text Available A simple, sensitive and specific stability indicating high performance liquid chromatographic (HPLC method for the estimation of Desloratadine was developed and validated. Desloratadine was separated and quantitated on Inertsil ODS-3 V column (250 mm length, 4.6 mm id, 5 μm particle size using a mixture of methanol–phosphate buffer of pH 7.0 (70:30 v/v as a mobile phase and at a flow rate of 1.0 mL/min. Quantification was achieved with an UV detector at 254 nm over the concentration range of 5–75 μg/mL. The applied HPLC method allowed the separation and quantification of Desloratadine with good linearity (r2 = 0.999 in the studied concentration range. Limit of detection and limit of quantification were found to be 1.28 μg/mL and 3.89 μg/mL, respectively. The method was validated as per the International Conference on Harmonization (ICH guidelines. Desloratadine stock solution was subjected to different stress conditions. The degraded product peaks were well resolved from the pure drug peak with significant difference in their retention time values. Stressed samples were assayed using developed HPLC method. Statistical analysis of the data showed that the method is precise, accurate, reproducible, and selective for the analysis of Desloratadine. The method was successfully applied to the estimation of Desloratadine in tablet dosage form.

  17. Chromatographic behavior of small organic compounds in low-temperature high-performance liquid chromatography using liquid carbon dioxide as the mobile phase.

    Science.gov (United States)

    Motono, Tomohiro; Nagai, Takashi; Kitagawa, Shinya; Ohtani, Hajime

    2015-07-01

    Low-temperature high-performance liquid chromatography, in which a loop injector, column, and detection cell were refrigerated at -35ºC, using liquid carbon dioxide as the mobile phase was developed. Small organic compounds (polyaromatic hydrocarbons, alkylbenzenes, and quinones) were separated by low-temperature high-performance liquid chromatography at temperatures from -35 to -5ºC. The combination of liquid carbon dioxide mobile phase with an octadecyl-silica (C18 ) column provided reversed phase mode separation, and a bare silica-gel column resulted in normal phase mode separation. In both the cases, nonlinear behavior at approximately -15ºC was found in the relationship between the temperature and the retention factors of the analytes (van't Hoff plots). In contrast to general trends in high-performance liquid chromatography, the decrease in temperature enhanced the separation efficiency of both the columns.

  18. Incorporation of ionic liquid into porous polymer monoliths to enhance the separation of small molecules in reversed-phase high-performance liquid chromatography.

    Science.gov (United States)

    Wang, Jiafei; Bai, Ligai; Wei, Zhen; Qin, Junxiao; Ma, Yamin; Liu, Haiyan

    2015-06-01

    An ionic liquid was incorporated into the porous polymer monoliths to afford stationary phases with enhanced chromatographic performance for small molecules in reversed-phase high-performance liquid chromatography. The effect of the ionic liquid in the polymerization mixture on the performance of the monoliths was studied in detail. While monoliths without ionic liquid exhibited poor resolution and low efficiency, the addition of ionic liquid to the polymerization mixture provides highly increased resolution and high efficiency. The chromatographic performances of the monoliths were demonstrated by the separations of various small molecules including aromatic hydrocarbons, isomers, and homologues using a binary polar mobile phase. The present column efficiency reached 27 000 plates/m, which showed that the ionic liquid monoliths are alternative stationary phases in the separation of small molecules by high-performance liquid chromatography.

  19. 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.

  20. 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

    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. 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. 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. 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.

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

    Directory of Open Access Journals (Sweden)

    Abid Kamal

    2015-01-01

    Full Text Available 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.

  2. [Study on fluorescence labeling and determination of polypeptide (PC2~PC6) by high performance liquid chromatography].

    Science.gov (United States)

    Li, Jing-xi; Gao, Li-jie; Cao, Wei; Zheng, Li; Chen, Jun-hui; Xu, Xiu-li; Wang, Xiao-ru

    2014-12-01

    This study was based on the thiol groups (-SH) of PC2~PC6, which could be reacted with the Monobromobimane (mBBr), in order to get polypeptide derivatives with fluorescent signal. A new method was developed for measuring the Polypeptides by high performance liquid chromatography with fluorescence detector, then the chromatographic conditions of HPLC was optimized; meawhile the reaction proportion of PCs and mBBr was identified by Trap-MS. The results showed that, the reaction proportion of PCs and mBBr was 1:1, the polypeptide derivatives had good stability; the five compounds separation was better, and the peak time focused on the 16.6~22.0 min; the linear correlation coefficient of PC2, PC3, PC4, PC5 and PC6 was >0.9991, and the limits of quantification were 0.3, 0.05, 0.3, 0.5 and 0.8 mg · L(-1) respectively, the recovery rate was 83.0%-102.0%; the method was reproducible, RSD<2%, this method for measuring the peptide compounds was rapid and accurate.

  3. [Determination of alpha-arbutin, beta-arbutin and niacinamide in cosmetics by high performance liquid chromatography].

    Science.gov (United States)

    Cheng, Peng; Chen, Meilan; Zhu, Yan

    2010-01-01

    A high performance liquid chromatography (HPLC) method for the determination of two optical isomers of arbutin (alpha-arbutin and beta-arbutin) and niacinamide in cosmetics was developed. The samples were extracted by the mixture of salt water and chloroform (2:1, v/v). The separation was performed on an ODS-BP column (200 mm x 4.6 mm, 5 microm, Elite) with methanol-water (10:90, v/v) as the mobile phase at a flow rate of 0.5 mL/min and 25 degrees C. The detection wavelength was set at 220 nm and the sample injection volume was 20 microL. There were good linear relationships between the mass concentration and the peak areas of alpha-arbutin, beta-arbutin and niacinamide in the ranges of 0.07-50, 0.06-50 and 0.05-50 mg/L, respectively. The relative standard deviations (RSDs, n = 7) of alpha-arbutin, beta-arbutin and niacinamide were 1.65%, 1.73% and 1.33%, respectively. The proposed method has been applied for the determination of alpha-arbutin, beta-arbutin and niacinamide in cosmetics with recoveries of 91.7%-109.6%. This method is rapid, simple and suitable for the detection of whitening ingredients in cosmetic.

  4. Optimized high performance liquid chromatography-ultraviolet detection method using core-shell particles for the therapeutic monitoring of methotrexate$

    Institute of Scientific and Technical Information of China (English)

    Milagros Montemurro; María M. De Zan n; Juan C. Robles

    2016-01-01

    Methotrexate (MTX) is an antineoplastic drug, and due to its high toxicity, the therapeutic drug mon-itoring is strictly conducted in the clinical practice. The chemometric optimization and validation of a high performance liquid chromatography (HPLC) method using core–shell particles is presented for the determination of MTX in plasma during therapeutic monitoring. Experimental design and response surface methodology (RSM) were applied for the optimization of the chromatographic system and the analyte extraction step. A Poroshell 120 EC-C18 (3.0 mm ? 75 mm, 2.7μm) column was used to obtain a fast and efficient separation in a complete run time of 4 min. The optimum conditions for the chroma-tographic system resulted in a mobile phase consisting of acetic acid/sodium acetate buffer solution (85.0 mM, pH¼4.00) and 11.2%of acetonitrile at a flow rate of 0.4 mL/min. Selectivity, linearity, accuracy and precision were demonstrated in a range of 0.10–6.0 mM of MTX. The application of the optimized method required only 150 mL of patient plasma and a low consumption of solvent to provide rapid re-sults.

  5. A VALIDATED REVERSE PHASE HIGH PERFORMANCE LIQUID CHROMATOGRAPHIC METHOD FOR THE DETERMINATION OF DOXOFYLINE IN PURE AND PHARMACEUTICAL FORMULATIONS

    Directory of Open Access Journals (Sweden)

    Lanka Venkateswararao

    2013-02-01

    Full Text Available A novel reverse phase high performance liquid chromatographic method (RP-HPLC was developed and validated for the determination of doxofylline (DXF in pharmaceutical formulations. Agilent 1100 series HPLC instrument equipped with variable wavelength programmable UV-Visible detector and a chromosil C18 (250mm x 4.6mm, 5μm column with an auto injector was used for the present investigation. A volume of 20µL of working standard of concentration 25µg/mL was injected into the chromatographic system, the component was separated by using the mobile phase of the mixture of potassium dihydrogen phosphate buffer of pH 5.5 and acetonitrile in the ratio 75:25(v/v at a flow rate of 1.0 mL/min and the detection of the components was carried out at a wavelength of 275nm. Chemstation software was used for the data acquisition through out the analysis. The system suitable parameters such as number of theoretical plates, tailing factor and resolution were found to be satisfactory. The retention time of the component was found to be at 4.814 minutes. The proposed method was validated in terms of precision, accuracy, linearity, limit of detection, limit of quantification, robustness and ruggedness. The developed method was found to be rapid, simple and sensitive hence it could be used as an alternative method in assay of the doxofylline in any pharmaceutical industries.

  6. Simultaneous determination of niflumic acid and its prodrug, talniflumate in human plasma by high performance liquid chromatography.

    Science.gov (United States)

    Jang, Dong-Jin; Park, Jeong-Sook; Ko, Hye-Ran; Jee, Jun-Pil; Kim, Jin-Ki; Kim, Sung Tae; Kim, Chong-Kook

    2005-01-01

    A high-performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of niflumic acid and its prodrug, talniflumate, in human plasma. Niflumic acid and talniflumate were eluted isocratically with methanol-water (73:27, v/v, adjusted to pH 3.5 by acetic acid) at a fl ow rate of 1 mL/min. Indomethacin was used as an internal standard. Signals were monitored by an UV detector at 288 nm. Retention times of indomethacin, niflumic acid and talniflumate were 5.9, 7.2 and 13.5 min, respectively. Calibration plots were linear over the range 50-5000 ng/mL for niflumic acid and 100-5000 ng/mL for talniflumate. The limits of quantitation were 50 ng/mL for niflumic acid and 100 ng/mL for talniflumate. The intra- and inter-day relative standard deviations (RSD) of niflumic acid and talniflumate were less than 10% and the accuracies were higher than 90%. This method is rapid, sensitive and reproducible for the determination of niflumic acid and talniflumate in human plasma.

  7. [Simultaneous determination of canthaxanthin and Sudan Red in salted duck egg yolk by reversed-phase high performance liquid chromatography].

    Science.gov (United States)

    Wang, Quanlin; Shi, Pingping; Zhang, Shufen; Shen, Jian; Fu, Xiao

    2007-11-01

    A reversed-phase high performance liquid chromatographic (RP-HPLC) method for the simultaneous determination of canthaxanthin, Sudan Red I , Sudan Red II , Sudan Red III and Sudan Red IV in salted duck egg yolk has been developed. These canthaxanthin and Sudan Red dyes were extracted with a mixture of acetonitrile, methanol, and chloroform (1 : 0.5 : 0.5, v/v). The extract was dried by a rotary evaporator. The residue was transferred to a graduated tube and diluted to 10 mL with acetonitrile. The canthaxanthin and Sudan Red dyes were separated on a XDB-C18 column (250 mm x 4.6 mm), with acetonitrile-water (95:5, v/v) as mobile phase. They were then detected at 478 nm - 520 nm - 471 nm at different retention times by UV/Vis spectrophotometer. The recoveries of sudan I , sudan II, sudan II, sudan IV, and canthaxanthin were 97.34%, 89.56%, 90.98%, 93.63% and 95.15% respectively. The relative standard deviations (RSDs) were 4.7%, 4.3%, 5.1%, 4.9% and 3.1%, respectively. The method is simple, rapid and accurate.

  8. Separation and purification of uridine diphosphate-glucuronosyltransferases by chromatofocusing on a high-performance liquid chromatograph.

    Science.gov (United States)

    Takanashi, H; Homma, H; Matsui, M

    1989-06-01

    A rapid method for the separation and purification of uridine diphosphate-glucuronosyltransferases (GT) was developed with the use of chromatofocusing on a high-performance liquid chromatograph. GT isoenzymes solubilized from hepatic microsomes of Wistar rats were separated on a Mono P column, a pre-packed column for chromatofocusing. Using 4-nitrophenol, testosterone and androsterone as substrates, four fractions with different GT activities were separated in a pH gradient from 9.5 to 7.0. Two isoenzymes, testosterone GT and androsterone GT were purified to apparent homogeneity. They were eluted at pH 8.9 and 8.0 and had subunit molecular weight values of 50000 and 52000, respectively. Approximately 10 mg of solubilized microsomal proteins was applied and the elution was completed within 2 h. Addition of N-nitrodiethylamine, an in vitro activator of GT activity, enhanced the GT activity toward 4-nitrophenol in the three fractions. This chromatographic analysis confirmed the absence of androsterone GT isoenzyme in LA Wistar rats, a mutant strain in terms of androsterone glucuronidation.

  9. High Performance Liquid Chromatography for the Determina- tion of Metoclopramide in Pharmaceutical Preparations Using Pre-column Derivatization with Fluorescamine

    Institute of Scientific and Technical Information of China (English)

    Tatar Ulu, Sevgi

    2012-01-01

    A simple, rapid and sensitive high performance liquid chromatography (HPLC) method was developed for the determination of metoclopramide in pharmaceutical preparation. The method is based on the derivatization of metoclopramide with fluorescamine. The separation was achieved on a C18 column using methanol-water (70 : 30, V/V) mobile phase. Fluorescence detector was used at the excitation and emission of 403 and 485 nm, respectively. The method was validated for linearity, limit of detection, limit of quantification, precision, accuracy, recovery, robust- ness and system suitability. The assay was linear over the concentration range of 100-2000 ng/mL. The mean recovery was 100.37%. The proposed method was successfully applied to the assay of metoclopramide in tablet preparation. The preparation was also analyzed with an official method and statistical comparison by t- and F-tests revealed that there was no significant difference between the results of the two methods with respect to mean values and standard deviations at the 95% confidence level.

  10. Determination of methylmercury in the tuna tissue by High performance liquid chromatography with Inductively Coupled Plasma Mass Spectrometry

    Institute of Scientific and Technical Information of China (English)

    LV Lei; Jiang Zhi-gang; Li Qun; Zhang Shuai

    2009-01-01

    This study mainly applied one kind oftechnologies to determine the methylmercury in the tuna tissue with high performance liquid chromagraphy-inductively coupled plasma nlass spectrometry.We choosed the optmaum conditiom including that the mobile phase Was water with 5% methanol,2-mercaptoethanol,0.06mol/Lammomum acetate and pH6.8.The sample amount was 1OOuL and the flow rate was 0.4mL/min.The optimum checking conditions is that RF power was 1500W,integral time was 0.3s sampling depth Was 8.0mm,velocity of carrier gas Was 0.75L/min and velocity of auxiliary gas Was 0.40L/min.In this con~tiom.the calibration CUrVe was linear when the concentra tions od methylmercury was in the range of 0-4ug/mL(r=0.9996),the detection limit can achieve 9ng/L,RSD is 8.2%,and the addition recovery is 75%-97%.The results show this method is simple,rapid and sensitive and can be used for the daily analysis of tuna samples.

  11. Simultaneous Determination of Epinephrine, Noradrenaline and Dopamine in Human Serum Samples by High Performance Liquid Chromatography with Chemiluminescence Detection

    Institute of Scientific and Technical Information of China (English)

    CHEN, Fu-Nan; ZHANG, Ying-Xue; ZHANG, Zhu-Jun

    2007-01-01

    A simple, rapid and accurate high performance liquid chromatographic (HPLC) technique coupled with chemiluminescence (CL) detection was developed for the simultaneous determination of epinephrine (E),noradrenaline (NA) and dopamine (DA). It was based on the analyte enhancement effect on the CL reaction between luminol and potassium ferricyanide. The effects of various parameters, such as potassium ferricyanide concentration, luminol concentration, pH value and component of the mobile phase on chromatographic behaviors of the analytes (E, NA and DA) were investigated. The separation was carried out on C18 column using the mobile tions, E, NA and DA showed good linear relationships in the range of 1×10-8-5×10-6, 5.0×10-9-1.0× 10-6 and 5.0×10-9-1.0×10-6 g/mL respectively. The detection limits for E, NA and DA were 4.0×10-9, 1.0×10-9 and 8.0×10-10 g/mL. The proposed method has been applied successfully to the analysis of E, NA and DA in human serum samples.

  12. Determination of Flavonoids and Anthocyanins in Nitraria tangutorum by High Performance Liquid Chromatography Coupled with Tandem Mass Spectrometry.

    Science.gov (United States)

    Zhe, Gao; Ying-Chun, Wang; Yan-Xu, Chang

    2016-01-01

    Using high-performance liquid chromatography coupled with diode array detection and electrospray ionization tandem mass spectrometry (HPLC-DAD-MSn) method, qualitative and quantitative analysis of flavonoids of stems, leaves, fruits and seeds, and anthocyanidin of fresh fruits in Nitraria tangutorum were performed. A total of 14 flavonoid components were identified from the seeds of N. tangutorum including three quercetin derivatives, three kaempferol derivatives, and eight isorhamnetin derivatives. A total of 12, 10, and 7 flavonoid components were identified from leaves, stems, and fruits of N. tangutorum, respectively; all were present in seeds also. The total content of flavonoids in leaves was the highest, up to 42.43 mg/g·dry weight. A total of 12 anthocyanidin components were identified from the fresh fruits of N. tangutorum, belonging to five anthocyanidin. The total content of anthocyanidin in fresh fruits was up to 45.83 mg/100 g· fresh weight, of which the acylated anthocyanidin accounted for 65.7%. The HPLC-DAD-MS(n) method can be operated easily, rapidly, and accurately, and is feasible for qualitative and quantitative analysis of flavone glycosides in N. tangutorum.

  13. Optimisation of the separation of herbicides by linear gradient high performance liquid chromatography utilising artificial neural networks.

    Science.gov (United States)

    Tran, Anh T K; Hyne, Ross V; Pablo, Fleur; Day, W Roy; Doble, P

    2007-02-28

    An artificial neural network (ANN) was employed to model the chromatographic response surface for the linear gradient separation of 10 herbicides that are commonly detected in storm run-off water in agricultural catchments. The herbicides (dicamba, simazine, 2,4-D, MCPA, triclopyr, atrazine, diuron, clomazone, bensulfuron-methyl and metolachlor) were separated using reverse phase high performance liquid chromatography and detected with a photodiode array detector. The ANN was trained using the pH of the mobile phase and the slope of the acetonitrile/water gradient as input variables. A total of nine experiments were required to generate sufficient data to train the ANN to accurately describe the retention times of each of the herbicides within a defined experimental space of mobile phase pH range 3.0-4.8 and linear gradient slope 1-4% acetonitrile/min. The modelled chromatographic response surface was then used to determine the optimum separation within the experimental space. This approach allowed the rapid determination of experimental conditions for baseline resolution of all 10 herbicides. Illustrative examples of determination of these components in Milli-Q water, Sydney mains water and natural water samples spiked at 0.5-1mug/L are shown. Recoveries were over 70% for solid-phase extraction using Waters Oasis((R)) HLB 6cm(3) cartridges.

  14. Simultaneous determination of quercetin, kaempferol and isorhamnetin accumulated human breast cancer cells, by high-performance liquid chromatography.

    Science.gov (United States)

    Wang, Yi; Cao, Jiang; Weng, Jian-Hua; Zeng, Su

    2005-09-01

    Quercetin, kaempferol and isorhamnetin are the most important constituents in ginkgo flavonoids. A simple, rapid and sensitive high-performance liquid chromatography method was developed to simultaneously determine quercetin, kaempferol and isorhamnetin absorped by human breast cancer cells. Cells were treated with ginkgo flavonols and then lysed with Triton-X 100. The flavonols in the samples were measured by RP-HPLC with a C18 column after a simple extraction with a mixture of ether and acetone. The mobile phase contained phosphate buffer (pH 2.0; 10 mM) tetrahydrofuran, methanol and isopropanol (65:15:10:20, v/v/v/v). The ultraviolet detector was operated at 380 nm. The calibration curve was linear from 0.1 to 1.0 microM (r > 0.999) for each flavonol. The mean extraction efficiency was about 70%. The recovery of the assay was between 98.9 and 100.6%. The limit of detection was 0.01 microM for quercetin and kaempferol and 0.05 microM for isorhamnetin. The limit of quantitation was 0.1 microM (R.S.D.isorhamnetin in human breast cancer Bcap37 and Bcap37/MDR1 cells.

  15. Phytochemical profile and nutraceutical potential of chia seeds (Salvia hispanica L.) by ultra high performance liquid chromatography.

    Science.gov (United States)

    Martínez-Cruz, Oliviert; Paredes-López, Octavio

    2014-06-13

    Chia seeds (Salvia hispanica) were analyzed for total phenolic compounds, antioxidant activity, and quantification of phenolic acids and isoflavones by ultra high performance liquid chromatography (UHPLC), in order to obtain a phenolic phytochemical profile. The total phenolic concentration was 1.8-fold higher than previous reports and the antioxidant activity using DPPH (2,2-diphenyl-1-picrylhydrazyl) radical assay showed 68.83% inhibition, which was higher than the values reported previously for chia and different plant foods. Additionally, a simple, reproducible and rapid UHPLC method was proposed for the analysis of phenolic acids and isoflavones in chia. The method demonstrated to perform well with regard to linearity, limits of detection and quantification, precision, accuracy, and sensitivity. The detection limits ranged from 0.05 to 0.4ng/mL and the recovery percentage from 23.62 to 162.48%. With this method the major compounds identified and quantified were: rosmarinic acid 0.92, protocatechuic ethyl ester 0.74, caffeic acid 0.02, gallic acid 0.01, and daidzin 0.006mg/g seed. In brief, this study demonstrates that chia could be considered a seed with high antioxidant capacity and novel isoflavone source that can be incorporated in human diet.

  16. [Determination of apomorphine, sildenafil and alprostadil in medicines for erectile dysfunction by high performance liquid chromatography-mass spectrometry].

    Science.gov (United States)

    Xu, Yuanjin; Xu, Guiping

    2005-11-01

    A high performance liquid chromatography-mass spectrometry (LC-MS) analytical method for illicit drugs, apomorphine, sildenafil and alprostadil, in medicines for erectile dysfunction has been developed. The samples were extracted with methanol using ultrasound-assisted extraction. The chromatographic separation was performed on a Zorbax Eclipse XDB-C18 column using acetonitrile-0.5% formic acid aqueous solution as mobile phase. The three compounds were identified by retention time and m/z and quantified by peak area. The results demonstrated that the linear ranges were 50.0 - 5 000.0 microg/L, 10.0 - 1 000.0 microg/L, 40.0 - 4 000.0 microg/L, with detection limits of 20.0, 4.0, 10.0 microg/L for apomorphine, sildenafil and alprostadil, respectively. The average recoveries and the relative standard deviations were 89% - 95% and 9.5% - 11%. The method is simple, rapid, accurate and suitable for the simultaneous determination of these drugs in medicines for erectile dysfunction.

  17. [Determination of phthalate plasticizers in foods by high performance liquid chromatography with gel permeation chromatographic clean-up].

    Science.gov (United States)

    Zhang, Chunyu; Wang, Hui; Zhang, Xiaohui; Ma, Zhongqiang; Deng, Wanmei; Hu, Ke; Ding, Mingyu

    2011-12-01

    A method of gel permeation chromatography-high performance liquid chromatography (GPC-HPLC) was established for the simultaneous determination of 5 main phthalate plasticizers in foods (edible oil, instant noodles, fried pastries, Saqima, etc.). The samples were extracted with petroleum ether in an ultrasonator, purified by a GPC column, and analyzed by HPLC. The chromatographic separation was achieved on a Labtech-C18 column (250 mm x 4.6 mm, 5 microm) using acetonitrile and water mixture as the mobile phases in a gradient elution mode. The developed method exhibited a linear correlation coefficient of more than 0.997 and the detection limits of 3.25 - 13.4 microg/L. The spike recoveries were between 70.4% and 113.6% with the relative standard deviations (RSDs, n = 3) of 0.3% - 5.8% at the spiked level of 50 mg/L. This method is simple, rapid and practical, and can be used for the simultaneous determination of PAEs in grease food samples.

  18. Improved proteomic discovery by sample pre-fractionation using dual-column ion-exchange high performance liquid chromatography.

    Science.gov (United States)

    Havugimana, Pierre C; Wong, Peter; Emili, Andrew

    2007-02-15

    Clinically relevant biomarkers are urgently needed for improving patient diagnosis, risk stratification, prognosis and therapeutic treatments. There is a particularly compelling motivation for identifying protein-based indicators of early-stage disease for more effective interventions. Despite recent progress, the proteomic discovery process remains a daunting challenge due to the sheer heterogeneity and skewed protein abundances in biofluids. Even the most advanced mass spectrometry systems exhibit limiting overall dynamic ranges and sensitivities relative to the needs of modern biomedical applications. To this end, we report the development of a robust, rapid, and reproducible high performance ion-exchange liquid chromatography pre-fractionation method that allows for improved proteomic detection coverage of complex biological specimens using basic tandem mass spectrometry screening procedures. This form of sample simplification prior to global proteomic profiling, which we refer to collectively as 'fractionomics', increases the number and diversity of proteins that can be confidently identified in tissue and cell lysates as compared to the straight analysis of unfractionated crude extracts.

  19. [Determination of amygdalin content in Semen Armeniacae Amarum by high-performance liquid chromatography with evaporative light-scattering detection].

    Science.gov (United States)

    Wu, Zhao-hui; You, Wen-wei; He, Feng

    2005-12-01

    High-performance liquid chromatography with evaporative light-scattering detection (HPLC-ELSD) was employed for determination of amygdalin content in Semen Armeniacae Amarum. The detection was performed with the column of Hypersil-ODS (4.6 mm x 250 mm, 5 microm) and column temperature of 25 degrees C. The mobile phase was methanol-water (70:30) with flow rate of 0.5 ml/min. Evaporative light-scattering detector was used and the drift tube temperature was set at 98 degrees Celsius with the gas flow rate of 3.2 L/min. A standard curve was generated, which was linear in the range of 1.0-15.1 microg for amygdalin content (r=0.999 9). The average recovery of amygdalin was 99.0% with RSD of 2.9% (n=5). Besides simplicity and rapidness, the method yields accurate and reproducible results and can therefore be used in the quality control of Semen Armeniacae Amarum.

  20. 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 ...