Development of validated stability indicating assay method for the simultaneous estimation of hydrochlorothiazide Amlodipine besylate and Losartan potassium in combine dosage form

Abstract

A Stability indicating Reverse-Phase liquid chromatographic method for the simultaneous estimation of HCTZ, LOSA and AMLO was developed. The chromatographic assay involves the use of SUPELCO LC-8-DB column (15 cm x 4.6 mm, 5 m) with a simple mobile phase composition of Buffer (monobasic Potassium Dihydrogen phosphate of 0.025 M having pH 3.7): Acetonitrile (60:40) at a flow rate of 1mL/min with U.V detection at wavelength of 232 nm. The method showed good linearity in the concentration range of 4-40 ?g/mL for HCTZ and 2-22 ?g/mL for AMLO and 15-150 ?g/mL for LOSA. The proposed method was also successfully applied to 20 tablets of marketed formulation (Trilopace). The developed method was successfully validated as per the ICH guidelines for following parameters. Accuracy, precision, ruggedness, robustness, system suitability tests, etc. The RSD for system precision was found to be 0.89-0.49 for HCTZ, AMLO, and LOSA and for method precision 1.0-1.4 for HCTZ, AMLO, and LOSA. The average percentage recoveries 99.75, 99.88, 98.93 for HCTZ, AMLO, LOSA which was in good agreement with labeled amount of Pharmaceutical formulation. The stability indicating capacity was tested by accelerated degradation of marketed formulation in acidic (0.1 N HCL), basic (0.1 N NaOH), , Oxidative (3% H ₂ O ₂ ), Thermal (80 ⁰ C)