HPTLC Method for Simultaneous Estimation of Lamivudine, Tenofovir Disoproxil Fumarate, and Doravirine

Abstract

Objective: The objective of this study was to develop and validate an HPTLC method for the simultaneous estimation of Lamivudine, Tenofovir disoproxil fumarate, and doravirine. The method is aimed to provide reliable and efficient quantification of these drugs. Methods: The chromatographic separation of drugs was performed on aluminum plates coated with silica gel 60 F254. Samples were spotted on the plate as a 6 mm wide band using a linomat applicator and a 100 µl syringe. The mobile phase used was a mixture of ethyl acetate, methanol, and chloroform (07:02:01 % v/v/v). Densitometric scanning at 226 nm was conducted using a Deuterium lamp as the radiation source, and the data were analyzed using win CATS software. The method was validated following the ICH Guideline ICH Q2 (R1). Results: The optimized method lead to the resolution of drugs with the Rf values of doravirine (0.75±0.02), Tenofovir disoproxil fumarate (0.57±0.02), and lamivudine (0.37±0.02). Doravirine exhibited a linear range of 500-1500 ng/band with a favorable linear equation and regression coefficient of 0.999. Tenofovir disoproxil fumarate and lamivudine showed a linear range of 1500-4500 ng/band, and both compounds displayed a linear relationship with a regression coefficient of 0.997. The method's accuracy was assessed through recovery studies, and the LOD and LOQ were determined for each drug. Conclusion: The optimized HPTLC method was validated in this study, following the ICH Q2 (R1) guidelines, it demonstrates its efficacy for the quantitative analysis of Doravirine, Tenofovir disoproxil fumarate, and lamivudine. The method offers reliable quantification of these compounds in a combined dosage form and can be used for routine analysis in pharmaceuticals.