High performance liquid chromatography was coupled with UV detection for simultaneous quantification of lopinavir （LPV） and ritonavir （RTV） in human plasma. This assay was sensitive, accurate and simple, and only ...High performance liquid chromatography was coupled with UV detection for simultaneous quantification of lopinavir （LPV） and ritonavir （RTV） in human plasma. This assay was sensitive, accurate and simple, and only used 200μL of plasma sample. Samples were liquid-liquid extracted, and diazepam was used as an internal standard. The chromatographic separation was achieved on a C18 reversed-phase analytic column with a mobile phase of acetonitrile-sodium dihydrogen phosphate buffer （10 mmol L-1, pH 4.80） （60：40, v/v）. UV detection was conducted at 205 nm and the column oven was set at 40℃. Calibration curves were constructed between 0,5-20 μg mL-1 for LPV and 0.05-5 μg mL-1 for RTV. The relative standard deviations were 2.16%-3.20% for LPV and 2.12%-2.60% for RTV for intra-day analysis, and 2.34%-4.04% for LPV and 0.31%-4.94% for RTV for inter-day analysis. The accuracy was within 100%＋10%. The mean extraction recoveries were 79.17%, 52.26% and 91.35% for RTV, LPV and diazepam, respectively. This method was successfully applied to human plasma samples from patients orally administered a salvage regimen of lopinavir-ritonavir tablets.展开更多
基金supported by the National Key Technologies R&D Program for the 11th Five-year Plan (Grant No. 2008ZX10001-006)the Key Clinical Program of the Ministry of Health 2010-2012
文摘High performance liquid chromatography was coupled with UV detection for simultaneous quantification of lopinavir （LPV） and ritonavir （RTV） in human plasma. This assay was sensitive, accurate and simple, and only used 200μL of plasma sample. Samples were liquid-liquid extracted, and diazepam was used as an internal standard. The chromatographic separation was achieved on a C18 reversed-phase analytic column with a mobile phase of acetonitrile-sodium dihydrogen phosphate buffer （10 mmol L-1, pH 4.80） （60：40, v/v）. UV detection was conducted at 205 nm and the column oven was set at 40℃. Calibration curves were constructed between 0,5-20 μg mL-1 for LPV and 0.05-5 μg mL-1 for RTV. The relative standard deviations were 2.16%-3.20% for LPV and 2.12%-2.60% for RTV for intra-day analysis, and 2.34%-4.04% for LPV and 0.31%-4.94% for RTV for inter-day analysis. The accuracy was within 100%＋10%. The mean extraction recoveries were 79.17%, 52.26% and 91.35% for RTV, LPV and diazepam, respectively. This method was successfully applied to human plasma samples from patients orally administered a salvage regimen of lopinavir-ritonavir tablets.
