High throughput LC–MS/MS method for simultaneous determination of zidovudine, lamivudine and nevirapine in human plasma

A selective and sensitive high performance liquid chromatography–tandem mass spectrometry method has been developed and validated for simultaneous determination of zidovudine (ZDV), lamivudine (3TC) and nevirapine (NVP) in human plasma. After Solid phase extraction (SPE), analytes and ISTDs were run on Peerless Basic C18 column with an injection volume of 3 μL and run time of 3.0 min. An isocratic mobile phase of 0.1% formic acid in water:methanol (15:85, v/v) was used with positive mass spectrometric detection. The method was validated over a concentration range of 5–1500 ng/mL for ZDV and 3TC and over the concentration range of 10–3000 ng/mL for NVP. The intraday and interday precision and accuracy across four validation runs were ranged from 1.6 to 10.1% and 93.8 to 110.8% respectively.