Analytical and Bioanalytical Method for Quantification of Pure Azilsartan, Not its Salts by RP-HPLC

Abstract

A simple, specific and accurate reverse phase liquid chromatographic method was developed for determination of Azilsartan in its tablet dosage form as well as from biological fluid like plasma. The determination was carried out on ODS Hypersil C18 column (4.6 mm x 250 mm 5µm) column using a mobile phase of acetonitrile: water (pH 4) [60: 40 %v/v]. The flow rate was 1 mL/min with detection at 249 nm. The linearity response of the HPLC system for Azilsartan was obtained over the range 1-64 µg/mL. Ambroxol (25µg/mL) was used as internal standard. Retention time for Azilsartan and ambroxol were found to be 3.74 and 1.84 minutes respectively. The correlation co-efficient (R2 value) for Azilsartan was found to be 0.992. The proposed method was successfully used for quantification of Azilsartan in Azilva tablets and plasma. The method was validated as per ICH Q2B (Analytical) and USFDA (Bio-analytical) guidelines. The results of analysis have been validated statistically and by recovery studies. The method is found useful for quantification of Azilsartan in marketed formulations as well as from biological fluids and can be applied for quantification of Azilsartan in preclinical and clinical studies.