Development and validation of an HPLC-UV method for simultaneous determination of zidovudine, lamivudine, and nevirapine in human plasma and its application to pharmacokinetic study in human volunteers

摘要: A simple, rapid, and sensitive high performance liquid chromatographic method with UV detection has been developed validated according to the FDA guidelines for quantitation of zidovudine (ZDV), lamivudine (LMV), nevirapine (NVR) in human plasma. The sample was prepared by simple liquid-liquid extraction. Chromatographic separation carried out a Hypersil BDS, C(18) column (250 mm × 4.6 mm; 5 µm particle size) mobile phase composition 0.1 M ammonium acetate buffer 0.5% acetic acid, v/v methanol (40:60, v/v) at flow rate 0.85 ml min(-1) where detector set 270 nm total run time 10 min which is very short simultaneous estimation three analytes linear over concentration range 50-3000, 50-2000 10-3000 ng ml(-1) lower limit quantifications (LLOQ) 50, 10 ng ml(-1) ZDV, LMV, NVR, respectively. Accuracy precision values both within-run between-run obtained from six different sets quality control (QC) samples along LLOQ analyzed separate occasions all ranged 94.47-99.71% 0.298-3.507%, Extraction recovery plasma above 90.16%. In stability tests, were stable during storage assay procedure. successfully applied quantitative determination pharmacokinetic study 12 healthy volunteers.