Simultaneous determination of six HIV protease inhibitors (amprenavir, indinavir, lopinavir, nelfinavir, ritonavir and saquinavir), the active metabolite of nelfinavir (M8) and non-nucleoside reverse transcriptase inhibitor (efavirenz) in human plasma by high-performance liquid chromatography

Abstract

We report the development of a simple, economical and reliable chromatographic method for the simultaneous determination of six HIV protease inhibitors (PIs; amprenavir, indinavir, lopinavir, nelfinavir, ritonavir and saquinavir), the active metabolite of nelfinavir (M8) and the non-nucleoside reverse transcriptase inhibitor (NNRTI; efavirenz) in human plasma. After extraction from plasma with an ethyl acetate-acetonitrile mixture, the analytes were separated on a phenyl column with a gradient of acetonitrile and phosphate solutions, and detected at three ultraviolet wavelengths. Calibration curves were linear over the range 0.025-15 microg/mL for saquinavir and 0.05-15 microg/mL for the other analytes. The accuracies ranged from -6.9% to +7.6%, and the intra-assay and inter-assay precisions were <9.2 and <11.8%, respectively. Our method, covering most of the PIs and NNRTIs currently used, facilitates ready therapeutic drug monitoring in hospital laboratories.