High-pressure liquid chromatographic (HPLC) determination of indomethacin in plasma.
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A simple and rapid determination method for nitazoxanide (NTZ), an antiprotozoal agent, was developed using reverse-phase HPLC and Ultra Performance Liquid Chromatography™ (UPLC™). Only six minutes gradient condition for NTZ analysis using UPLC was achieved. The mobile phase consisted of a mixture of phosphate buffer (pH 6.0) and acetonitrile. The repeatability (relative standard deviation (RSD), n = 6) and the correlation coefficient from linearity (the range from 80% to 120% of amount) were 0.25% and 0.9963 for UPLC and 0.15% and 0.9988 for HPLC, respectively. The quantitative values of NTZ in tablets were 103.2% for HPLC and 98.7% for UPLC. The RSDs of quantitative values of sample solution were calculated to be 4.06% to 4.64% for HPLC and 0.15% to 0.36% for UPLC.
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Objective: To establish a HPLC UV method for the determination of hydroxycamptothecin in rabbit plasma. Method: After acid treatment,plasma samples were precipitated with methanol and then centrifugated. 20μL of supernatant was injected and measured by HPLC UV method. The Chromatographic column was Lichrospher C 18 column:(250 mm×4.6 mm,5μm). Methanol-0.01 mol/L phosphate buffer(pH4.0) (60 ∶ 40) was served as mobile phase at 1.0mL/min. Detection wavelength was 384 nm. Results: The retention time of hydroxycamptothecin was 5.0min. A good linearity was shown in the concentration range of 50～5000 ng/mL.The lowest detection concentration was 25 ng/mL.The recovery was between 99.88% and 103.13%. The intra day RSD was less than 3 27% and the inter day RSD was less than 7.25%. Conclusion: This method was simple,practical and accurate. It could be applied in pharmacokinetic study of hydroxycamptothecin.
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Norisoboldine is an alkaloid and has been identified as the major active component in Linderae Radix. A novel method for quantitative analysis of norisoboldine in Linderae Radix using ultra performance liquid chromatography (UPLC) with UV detection was developed for quality control. A similar and conventional HPLC protocol was simultaneously developed for comparison purposes. Chromatographic separation of norisoboldine was performed on an Acquity UPLC BEH C18 column (50 mm × 2.1 mm ID, 1.7 µm) for UPLC and on a Waters X-Bridge C18 column (250 mm × 4.6 mm ID, 5.0 µm) for HPLC. In the UPLC method, the average recovery was 97.3% (n = 9, RSD 1.1%); the limit of detection (LOD) and limit of quantification (LOQ) were 0.151 and 0.302 ng, respectively. In the HPLC method, the average recovery was 98.9% (n = 9, RSD 1.7%); the LOD and LOQ were 1.51 and 3.02 ng, respectively. The UPLC method provided a shorter analysis time, a better sensitivity in detection and quantitation, and less solvent was used in comparing to the HPLC method. The amount of norisoboldine detected in each Linderae Radix sample from different sources were in good agreement in both HPLC and UPLC methods.
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OBJECTIVE To establish an high performance liquid chromatography (HPLC) assay for the determination of concerttration of vancomycin in human plasma using norvancomycin as internal standard.METHODS The pretreatment of the blood samples was simply sedimentation with 10% trichlomacetic acid.Then the samplewas separated on Agilent C18 column with a mixture of methanol-acetonitrile-0.05mol·L-1 potassium dihydrogenp hosphate(15∶2∶83,pH3.2).The UV wavelengthw as 236nm.The flow rate was 1.0 mL·min-1.The temperature of the column was 35℃.RESULTS A linearity was obtained from 1.3mg to 83.2mg·L-1 of vancomycin with good correlation(r0.999).The average recovery was 100.6%.The RSD for intra-day and inter-day assays were less than 4%.CONCLUSION It appears to be a rapid,convenient and accurate method for determination of vancomycin in plasma.
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Objective:To develop an HPLC method for determination of paraquat in rat plasma to provide evidence for the therapy and prognosis of paraquat-poisoned patients.Methods:The rat plasma samples were deproteimized with 35%(v/v) perchlorld acid.A Diamonsil TM C18 column (250 mm×4.6 mm,5 μm) with the mobile phase consisted of 0.1 mol·L-1 phosphate buffer (75 mmol·L-1 sodium heptanesulfonate)-acetonitrile (88∶12),adjusted pH to 3.0 by triethylamine was used to separate paraquat in biological samples,and the detection occurred at 258 nm.Result:A good linearity was obtained in the concentration range of 20-5000 ng·mL-1,the average recovery was 87.5%,the intra-day precision (RSD) was less than 10%.The concentration of paraquat in rat plasma was (360.3±130.9)ng·mL-1 after ig. 50 mg·kg-1 paraquat 8 hours,and paraquat couldn't be detected in rat plasma at the third day.Conclusion:The described method was proved to be sensitive,rapid and accurate,can be applied in identification and determination of paraquat in rat plasma.
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OBJECTIVE The pharmacokinetics of cucurbitacins in mice was studied, and the HPLC method was established for the separation and quantitative determination of cucurbitacins B in plasma.METHOD Column: Hypersil ODS C 18 (150 mm×4.6 mm, 5 μm); column temperature: 30℃;mobile phase: a mixture of methanol(MeOH) and water(72∶28); flow rate: 0.5 mL·min -1 ; detection wavelength: 228 nm; aliquot injected into the HPLC system: 20 μL.RESULTS The recovery of the assay was 94.11%～99.77% with the intra-day and inter-day RSD less than 12.12% and 15.63%,respectively.The detection limit was 4ng. The lowest detection concentration was 50 ng·mL -1 in plasma.CONCLUSON This method was found to be simple, rapid ,sensitive and accurate for the determination of CuCB in plasma.
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The purpose of these studies was to develop a high performance liquid Chromatographie (HPLC) assay for plasma levels of indomethacin.A reversed phase (C 18 Sep-Pak) cartridge was used to process plasma for absorption of indomethacin, the internal Standard, and impurities.The recovery of plasma indomethacin and the added internal Standard was quantitative from 0.5 ml of plasma.The assay was linear from 50 g/l to 10 mg/1 without concentration of the effluent from the Sep-Pak cartridge.The intra-assay coefficients of Variation, for ten injections each to calibrate points at 0.15, 0.30, 0.50 and 1.00 mg/1, were 8.15%, 6.29%, 5.47% and 5.39%, respectively and for duplicates of duplicate calibration points were 7.51%, 6.32%, 4.41%, and 2.05%, respectively.The inter-assay coefficients of Variation were 8.49%, 6.48%, 5.10%, and 2.22%, respectively.The sensitivity of the assay can be increased by a 3 -5 fold concentration of the effluent from the Sep-Pak and preliminary experiments have indicated that äs little äs 100 of starting plasma can be utilized in the assay.The assay can be used to determine the concentration of indomethacin in small volumes of plasma.Since Cig Sep-Pak cartridges were employed to remove contaminating substances, sensitivity and reproducibility were both high while column longevity and efficiency were excellent. Die Verwendung von Reversed Phase-Patronen (C 1S ) för die Aufarbeitung von Plasma zur Analyse von Indomethacin durch Hochleistuhgsflüssigchromatographie
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