Ultra-Performance Liquid Chromatographic Method for Simultaneous Quantification of HIV Non-Nucleoside Reverse Transcriptase Inhibitors and Protease Inhibitors in Human Plasma

A fast ultra-performance liquid chromatographic with diode-array detection method has been developed and validated for the determination of six protease inhibitors (indinavir, amprenavir, saquinavir, atazanavir, lopinavir, and ritonavir) and two non-nucleoside reverse transcriptase inhibitors (efavirenz and nevirapine). After liquid-liquid extraction of 0.5 mL plasma with methyl-tert-butyl ether, the analytes were separated on a ACQUITY UPLC BEH (R) C18 column (2.1 x 150 mm, p.d. 1.7 mu m) column eluted with a gradient of acetonitrile and triethylammonium phosphate buffer 5 mmol L-1 pH 3.0. The total run time was 9.5 min. Calibration curves were linear in the range 0.1 to to 10.0 mu g mL(-1). The lower limit of quantitation was 0.1 mu g mL(-1) for all drugs. Accuracy ranged from 94.9 to 103.5%. Both interday and intraday coefficients of variation were less than 7.7% for all analytes. The extraction yields were greater than 88.2%.