High-performance liquid chromatographic method for the simultaneous determination of HIV-1 protease inhibitors indinavir,saquinavir and ritonavir in plasma of patients during highly active antiretroviral therapy

摘要

S new high-performance liquid chromatographic method for the simultaneous determination of indinavir,saquinavir and ritonavir in human plasma is described. Quantitative recovery following liquid-liquid extraction with diethyl ether from 500 #mu3l of human plasma was achieved. Subsequently,the assay was performed with a linear gradient starting at 67 mM potassium dihydrogenphosphate-acetonitrile (65:35 to 40:60, v/v) as a mobile phase,a Phenomenex C_18 column and UV detection at 240 and 258 nm,respectively. Linear standard curves were obtained for concentrations ranging from 75 to 20 000 ng/ml for indinavir,from 10 to 6000 ng/ml for saquinavir, and from 45 to 30 000 ng/ml for ritonavir. The calculated intra- and inter-day coefficients of variation were below 6%.