DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ESTIMATION OF LAMIVUDINE, TENOFOVIR AND EFAVIRENZ BY UPLC

Sk. Madeesh, Y. Ismail and V. Gunasekaran

A simple, accurate, rapid and precise isocratic reversed-phase high-performance liquid chromatographic method has been developed and validated for simultaneous determination of lamivudine, tenofovir and efavirenz in tablets. The chromatographic separation was carried out on a BEH symmetry C18 (50×4.6mm, 1.7 μm) column with a mixture of methanol: phosphate buffer pH 3.0 adjusted with o-phosphoric acid (65:35, v/v) as mobile phase; at a flow rate of 0.3 mL/min. The retention times for LAM, TEN and EFA were observed to be 0.432, 0.657, 2.281 min, respectively. Calibration plots were linear (r 2 >0.999) over the concentration range of 10-50 μg/mL for LAM and TEN; and 20-100 μg/mL for EFA. The method was validated for accuracy, precision, specificity, linearity, and sensitivity. The proposed method was successfully used for quantitative analysis of tablets. No interference from any component of pharmaceutical dosage form was observed. Validation studies revealed that method is specific, rapid, reliable, and reproducible. The high recovery and low relative standard deviation confirm the suitability of the method for routine determination of lamivudine, tenofovir and efavirenz in tablets.