Original Articles: 2012 Vol: 4 Issue: 6
Determination of nevirapine in human plasma by HPLC
Nevirapine is a non–nucleoside Reverse Transcriptase inhibitor, antiviral agent. A simple, selective, and sensitive high performance liquid chromatography method for the determination of Nevirapine in human plasma was developed. Dicloxacillin was used as an internal standard. The method utilizes simple protein precipitation as the sample preparation technique using acetonitrile as precipitating agent. The samples were analyzed on HiQ sil C18 HS column with the mobile phase containing mixture of methanol: phosphate buffer 0.01M (pH 3.0, adjusted with glacial acetic acid) (70: 30% v/v). The calibration curve was linear (r2>0.99) through the range of 0.2-1.2µg/ml. The lower limit of quantification was found to be 0.2 µg/ml. % R.S.D. was less than 4% for intra- and inter-day precision. The mean recovery was found to be 96.50% for Nevirapine. Nevirapine in plasma samples was stable when tested for parameters as per EMEA and US CDER guidelines. The method showed acceptable values for accuracy, precision, recovery, sensitivity and stability. The method is well suited for routine analysis of Nevirapine in human plasma and can further be extended for pharmacokinetic studies.