Development of LC/LCMS method for estimation of impurities in anti-HIV drug (Bictegravir) using Analytical Quality by Design (AQbD) approach.

Abstract

Analytical Quality by Design (AQbD) was used to construct stability indicating linked compounds using the HPLC technique of an anti-HIV medicine (Bictegravir). Process-related and degrading impurities were separated on a Zorbax SB-C8 (150×4.6) mm, 3.5 m column. The buffer pH of the mobile phase was kept at 2.5 by employing potassium dihydrogen phosphate 0.05M. Mobile phase A contains 5% methanol and 95% buffer, whereas mobile phase B contains 50% acetonitrile, 10% methanol, 25% tetrahydrofuran, and 15% water. The completed chromatographic settings were a flow rate of 1.2 mL/min, a detector wavelength of UV at 250 nm, and an injection volume of 20 µL. This approach has been verified in accordance with ICH recommendations. The approach was discovered to be particular, sensitive, linear, exact, and accurate. The limit of quantification for all contaminants was determined to be less than 0.05%, the correlation coefficient for all impurities is between 0.9996 and 1.0000, and the percent recovery for all impurities is between 91% and 108% at the LOQ level and 97% to 113% at the specification level. Forced degradation experiments in chemical and physical stress tests were performed in accordance with regulatory criteria, and the molecule was discovered to be susceptible to acid and base hydrolysis. Imp-A was the most common degrading impurity in both acid and base hydrolysis.