An efficient RP-HPLC-PDA Method for Estimating Dolutegravir and Lamivudine in Combined Pharmaceutical Formulations using a Box-Behnken Design Approach

Abstract

This study used a quality-by-design (QbD) method to build a novel, high-performance reverse-phase fluid chromatography (RP-HPLC) system with diode array detection (DAD) for the simultaneous measurement of dolutegravir (DLG) and lamivudine (LMV) tablets. In accordance with the International Council for Harmonization’s (ICH) requirements, chromatographic conditions were adjusted utilizing the Box-Behnken design (BBD), and the resultant technique was verified for linearity, system appropriateness, precision, accuracy, sensitivity, robustness, and stability of the solution. At retention durations of 2,271 and 3,431 minutes, LMV and DLG peaks were separated using a C-18 column with 150 x 4.6 mm and 5 μm particles. The mobile phase was 0.1% orthophosphoric acid (OPA): acetonitrile (ACN) (50:50, v/v) at a flow rate of 0.8 mL/min and a pH of 3 at 25°C. Peaks were seen at 254 nm and the volume of sample injection was 20L. The percent strength of the commercially available tablet is 98.89 and 98.76 for LMV and DLG, respectively, using a standard calibration curve. The developed RP-HPLC technique’s stability is shown by the suggested RP-HPLC method’s capacity to identify LMV and DLG in the existence of their degradation products. As per ICH requirements, the findings of the validation parameters for linearity, system appropriateness, accuracy, precision, robustness, and sensitivity were all within acceptable limits. It is simple to use, accurate, efficient, and reasonably priced to employ the innovative RP-HPLC technology with BBD application for QbD.