Development and validation of stability-indicating HPLC method for simeltaneous determination of Lamivudine, Tenofovir, and Dolutegravir in bulk and their tablet dosage form

Abstract

ObjectiveA Simple, accurate, specific and rugged reverse phase liquid chromatographic method was developed for the simultaneous estimation of Lamivudine, Tenofovir, and Dolutegravir in bulk and tablet dosage form. MethodsA reverse phase gradient program has been developed to separate the all four active ingredients. The ingredients present in different concentrations and chromatographic behavior 0.05 M Phosphate buffer pH 6.2 ± 0.05 adjusted with dilute potassium hydroxide solution, Acetonitrile was used as mobile phase. A gradient programing has been done, on a reverse phase C18 column (250 × 4.6 mm, 5 micron) with a flow rate 1 mL/min, monitored at 260 nm. ResultsThe mean retention times of Lamivudine, Tenofovir, and Dolutegravir were found to be 2.8, 5.2 and 11.5 min respectively. Linearity of Lamivudine, Tenofovir, and Dolutegravir was found to be 27–162 μg/mL, 27–162 μg/mL and 4.5–28 μg/mL respectively. ConclusionThe proposed method was validated in terms of Linearity, Range, Accuracy, Precision, Specificity, Robustness and stability studies and the method is successfully applies to the estimation of Lamivudine, Tenofovir, and Dolutegravir in combined tablet dosage form.