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.