A New Validated RP-HPLC Method for the Estimation of Darunavir Ethanolate in Bulk and Tablets

Abstract

Objective: Darunavir ethanolate is the last United States Food and Drug Administration approved protease inhibitor (PI). The drug is used along with other PIs (ritonavir/cobicistat) for the effective management of human immunodeficiency virus (HIV) HIV-1 infection. Darunavir ethanolate exerts its action by binding noncompetitively to HIV protease enzyme. The main objective of the present research work was to develop a new profound and novel reverse-phase high-performance liquid chromatography (RP-HPLC) for the estimation of darunavir ethanolate. Methods: As per the guidelines of the Food and Drug Administration and International Council for harmonization, the method was validated. The HPLC analysis was performed on the waters 2695 equipped with symmetry C18 column 3.5 μm, 150 mm × 4.6 mm, with a mixture of acetonitrile:0.1% phosphate buffer (50:50 V/V) as the mobile phase, at the flow rate of 1 ml/min. The total run time was 5 min and the detection was performed at the wavelength (λ) of 262 nm. Results: The retention time for darunavir ethanolate was found to be 2.269 min. The standard curves were obtained with R2 0.9997 and linear at the concentration range of 8–120 μg/ml. The limit of quantitation and limit of detection for darunavir ethanolate were found to be 0.08 μg/ml and 0.8 μg/ml, respectively. The method’s accuracy was tested by percentage recovery tests and found to be 100.3%. The results obtained were within the accepted standards for linearity, accuracy, precision, specificity, and robustness. Conclusion: The proposed RP-HPLC method can be applied for the routine analytical estimation of darunavir ethanolate in bulk and tablet formulations.