Simple and rapid determination of nevirapine in human serum by reversed-phase high-performance liquid chromatography

Nevirapine is an antiretroviral agent belonging to the class of non-nucleos ide reverse transcriptase inhibitors. We describe a fast, simple isocratic reversed-phase high-performance liquid chromatography method with a 30-mm l ong column for assaying nevirapine in human serum. After deproteinization o f 200 mul serum samples with 50% trichloroacetic acid, the supernatant was injected into a reversed-phase C-18 column, using 10 mM phosphate buffer (p H 5)-acetonitrile (82:18, v/v) as the mobile phase. Peak detection was perf ormed at 240 nm. Nevirapine retention time was 2 min. The method was valida ted over 0.1-10 mug/ml and the assay was linear over this concentration ran ge (r(2)>0.998). Within- and between-day precisions were less than 5.4%. Th e lower limit of quantification was 0.1 mug/ml. Nevirapine in human serum s amples was stable for 2 days at 20-25 degreesC, 15 days at 4 degreesC and 3 months at -20 degreesC. (C) 2001 Elsevier Science B.V. All rights reserved .