Bio analytical method development and validation for simultaneous estimation of atazanavir and cobicistat by RP-HPLC in human plasma
A simple, speedy, sensitive, accurate and validated isocratic RP-HPLC/UV bio analytical method was developed for the concurrent quantitative estimation of atazanavir and cobicistat in human plasma along with darunavir as an internal standard. Drugs were extracted by simple protein precipitation procedure using acetonitrile for deprotonation. Quantification of atazanavir and cobicistat was achieved on analytical C18 column (250 mm x 4.6 mm; 5µm) using a mobile phase comprising of buffer and acetonitrile in the proportion of 65:35 (v/v), pumped with a flow rate of 1 ml/min. The eluents were detected at a wavelength of 240 nm and atazanavir, cobicistat and darunavir were detected at retention times of 5.92, 6.75 & 3.95min, respectively. Calibration plots for atazanavir and cobicistat in spiked plasma were linear in range of 0.3-10 & 0.15-5 µg/ml, respectively. The method was validated for various parameters as per standard guidelines and it was found to be convenient for successful bio analysis of atazanavir and cobicistat in human plasma.
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