Simultaneous method development and validation for Tenofovir AF, Emtricitabine, and Bictegravir by RP-UPLC

Abstract

A new, accurate RP-UPLC method has been developed and validated for the simultaneous quantification of Bictegravir (BCG), Tenofovir Alafenamide (TAF), and Emtricitabine (ETB) in pharmaceutical formulations. Chromatographic separation was achieved on an Acquity UPLC BEH C18 column (2.1 × 50 mm, 1.7 μm) using an isocratic mobile phase of 10 mM phosphate buffer (pH 3.0): acetonitrile (60:40, v/v) at a flow rate of 0.3 mL/min and detection at 260 nm. Retention times for ETB, BCG, and TAF were 2.3, 3.2, and 3.7 minutes, respectively. The method was validated per ICH Q2 (R1) guidelines and demonstrated excellent linearity (r > 0.999), precision (%RSD < 1%), and high recovery (99.8–100.6%) for all analytes. Forced degradation studies indicated the method is stability-indicating. The proposed UPLC method is suitable for routine quality control analysis of these drugs in fixed-dose combination tablets.