Validated Stability-Indicating Assay UHPLC Method for Simultaneous Estimation of Doravirine, Lamivudine and Tenofovir Disoproxil Fumarate in Pure Material and Pharmaceutical Matrix

Abstract

An efficient, robust, and specific stability-indicating UHPLC assay method was acquainted for simultaneous determination of antiretroviral agents doravirine (DOV), lamivudine (LVD), and tenofovir disoproxil fumarate (TDF). The determination of cited agents was performed on UPLC system using UPLC BEH C18 (150 mm × 2.1 mm) with 1.7 µm particle size column at ambient temperature using a solvent system in a proportion of (60:40 % v/v) acetonitrile: potassium dihydrogen orthophosphate buffer; pH 4.5 ± 0.2 was adjusted with 0.1 % OPA. The PDA detector was subjected to a wavelength of 235 nm. DOV, LVD, and TDF were experienced in various stress procedures of forced degradation experiments, as recommended by the ICH, to assess the established method's stability-indicating practicability. Using the optimized procedure, selected agents were successfully distinguished from the peaks of their respective degradation products. Furthermore, the calibration curves for DOV, LVD, and TDF in over wide concentration ranges of 2–12μg/mL, 5–30μg/mL, and 5–30μg/mL, respectively, with determination coefficients (r2) of 0.997, 0.9994, and 0.9994. The method validation assays had less than 2 % relative standard deviations for accuracy, precision, repeatability, and robustness.