Objective: The present study aimed to develop stress indicating reverse phase high-performance liquid chromatography (RP-HPLC) method for the estimation of Bictegravir, Emtricitabine and Tenofovir alafenamide in pharmaceutical dosage form and validated in accordance with ICH guidelines. Method: The optimized conditions for the developed RP-HPLC method are Kromasil C18 (250 X 4.6mm, 5µ ) column maintained at 30ºC with a mobile phase consisting of 0.01N Buffer (KH2PO4) and Acetonitrile in the

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... o 60:40%v/v on isocratic mode at flow rate 1.0ml/min. The sample was detected at 272nm. Results: The retention time of Bictegravir, Emtricitabine and Tenofovir alafenamide was found to be 2.9, 2.2 and 3.3min respectively. The developed method was validated for accuracy, precision, specificity, ruggedness, robustness and solution stability. The method obeyed Beer's law in the concentration range of 12.5µg/ml-75µg/ml for Bictegravir, 50µg/ml-300µg/ml for Emtricitabine and 6.25µg/ml -37.5µg/ml for Tenofovir alafenamide with a correlation coefficient of 0.999 for BIC, FTC and TAF respectively. Forced degradation studies were conducted by exposing the drug solution to various stress conditions such as acidic, basic, peroxide, neutral, photolytic and thermal conditions. The net degradation was found to be within the limits, indicating that the drug is stable in stressed conditions. Conclusion: The developed method for the estimation of Bictegravir, Emtricitabine and Tenofovir alafenamide can be utilized for the routine analysis of pharmaceutical dosage form. Keywords: Stress indicating, Method development, Bictegravir, Validation, RP-HPLC.