Determination of atazanavir and other antiretroviral drugs (indinavir, amprenavir, nelfinavir and its active metabolite M8, saquinavir, ritonavir, lopinavir, nevirapine and efavirenz) plasma levels by high performance liquid chromatography with UV detection

A global method is proposed for therapeutic drug monitoring of atazanavir, a novel protease inhibitor and of all other protease inhibitors (PI) and non nucleoside reverse transcriptase inhibitors (NNRTI) which are currently used to treat HIV patients. All drugs are extracted after a liquid-liquid extraction and separated on a Cl 8 column with a binary gradient elution except lopinavir which is separated without this gradient. The absorbance is measured at 259 nm except for lopinavir (205 nm) and nevirapine (320 nm). This method is specific, accurate, precise (the intra-day and inter-day imprecision and inaccuracy are lower than 15%) and the limits of quantitation (0.40 mg/L for nevirapine, 0.10 mg/L for indinavir, 0.10 mg/l for M8, 0.05 mg/L for amprenavir, 0.10 mg/L for nelfinavir, 0.10 mg/L for saquinavir, 0.10 mg/L for ritonavir, 0.10 mg/L for efavirenz, 0.10 mg/L for atazanavir and 0.20 mg/L for lopinavir) are consistent with trough plasma concentrations allowing to use this method for therapeutic drug monitoring of PI and NNRTI. (C) 2004 Elsevier B.V. All rights reserved.