Simultaneous Estimation of Atazanavir and Ritonavir in Combined Tablet Dosage Form

Abstract

Ritonavir is an HIV protease inhibitor that interferes with the reproductive cycle of HIV. Although it was initially developed as an independent antiviral agent, Atazanavir (formerly known as BMS-232632) is an antiretroviral drug of the protease inhibitor (PI) class. Like other antiretrovirals, it is used to treat infection of human immunodeficiency virus (HIV). Scan the standard solution and test solution on UV/Visible spectrophotometer, over the spectral range 200 to 400 nm. Use diluent as blank. The UV spectrum of the test solution should exhibit maxima at the same wavelength (±2 nm) as that of a standard solution ATV and RTV show reasonably good response at 279 and 240 nm, respectively in methanol as solvent. For mobile phase mixed 85 mL of Buffer and 15 mL of HPLC grade Methanol. Filtered through 0.45 µ Millipore nylon filter and degassed. Calibration curves were plotted over concentration range of 10 – 30 µg/mL for ATV and of 5 – 15 µg/mL for RTV. Mixed standard stock solutions which contains ATV (200 µg/mL) and RTV (100 µg/mL) was diluted as mentioned. Accuracy was determined in terms of % recovery. Recovery experiments were carried out in triplicate by spiking standard drug to previously analyzed samples of the tablets. Method precision of the instrument was established by repeatedly injecting six standard solutions of ATV (20 µg/mL) and RTV (10 µg/mL) under same conditions on the same day. Intermediate precision was evaluated in terms of intra-day and inter-day precision by analysing 3 different solutions 3 times on the same day and on different days over entire concentration range for both drugs. The solution stability for both drugs in the assay method were carried out by leaving test and standard solutions in tightly capped volumetric flasks at room temperature for 24 hrs. Sample preparation was injected in triplicate and chromatograms were recorded then responses for the analyte peaks were measured and all data recorded were found complies with results.

Keywords: Ritonavir, Atazanavir, Accuracy, Recovery, Method development, validation.