Simultaneous Estimation of Lamivudine, Stavudine and Nevirapine by RP-HPLC in Tablet Formulation

A simple, precise, reliable, rapid and reproducible reversed-phase high performance liquid chromatography (RP-HPLC) method was developed and validated for the simultaneous estimation of lamivudine (3TC), stavudine (D4T) and nevirapine (NVP) present in multicomponent dosage forms. Gradient chromatography using Inertsil-ODS-3 (C18) Column (5 μm, 250mm x 4.60mm) eluted with two mobile phase components: mobile phase A comprising of 0.1% OPA and mobile phase B (methanol) with a flow rate of 1.0 mL/min and a detection wavelength at 254nm. Parameters such as linearity, precision, accuracy, recovery, specificity and ruggedness are studied as reported in the International Conference on Harmonization guidelines. The retention times for lamivudine, stavudine and nevirapine were 2.6, 4.08, and 10.92 min respectively. The calibration curves were linear (r² > 0.9997 for all three compounds). The linearity range and percentage recoveries for 3TC, D4T and NVP are 30–150, 6–30, 40–200μgml⁻¹and 100.95, 107.46, 106.74% respectively. The correlation coefficients for all components are close to 1. The relative standard deviations for three replicate measurements in three concentrations of samples in tablets are always less than 2%.