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-nucleoside reverse transcriptase inhibitors. We describe a fast, simple isocratic reversed-phase high-performance liquid chromatography method with a 30-mm long column for assaying nevirapine in human serum. After deproteinization of 200 mul serum samples with 50% trichloroacetic acid, the supernatant was injected into a reversed-phase C-18 column, using 10 mM phosphate buffer (pH 5)-acetonitrile (82:18, v/v) as the mobile phase. Peak detection was performed at 240 nm. Nevirapine retention time was 2 min. The method was validated over 0.1-10 mug/ml and the assay was linear over this concentration range (r(2)>0.998). Within- and between-day precisions were less than 5.4%. The lower limit of quantification was 0.1 mug/ml. Nevirapine in human serum samples 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.