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Abstract

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING HPTLC METHOD FOR SIMULTANEOUS ESTIMATION OF ASPIRIN, CLOPIDOGREL BISULPHATE AND ROSUVASTATIN CALCIUM IN BULK AND PHARMACEUTICAL DOSAGE FORMULATION.

Priyanka Agrawal, Janhavi R. Rao* and Chaitali Dhale

ABSTRACT

A sensitive, accurate, and precise stability-indicating HPTLC method has been developed for the simultaneous estimation of Aspirin (ASP), Clopidogrel Bisulphate (CLOP) and Rosuvastatin Calcium (ROS) in bulk and pharmaceutical dosage formulation. The method employed Toluene: Ethyl Acetate: Methanol: Ammonia (6:3.5:1.5:0.2 v/v/v/v) as a mobile phase and Silica Gel G 60 F254 TLC plates as stationary phase. Detection was performed at 258 nm. The Rf Value of ASP, CLOP and ROS were found to be 0.22 ±0.02, 0.88 ±0.02 and 0.16 ±0.02 respectively. The method was validated in compliance with ICH Guideline for linearity, limit of detection (LOD), limit of quantification (LOQ), precision, specificity, accuracy and robustness. The linear regression analysis (n=3) showed good linear relationship over the concentration range of 225 - 1350 ng/spot for ASP (r2=0.9999), 225 – 1350ng /spot for CLOP (r2=0.9991) and 90 – 180 ng/spot of ROS (r2=0.9992). The LOD of ASP, CLOP and ROS were found to be 1.671ng/spot, 2.211ng/spot and 0.207ng/spot respectively. The LOQ of ASP, CLOP and ROS were found to be 50.65ng/ spot, 67.01ng/spot and 6.27ng/spot respectively. The % recovery was calculated and found to be 99.27– 100.4% for ASP, 98.2%-99.17% for CLOP and 98.86-101.15% for ROS. ASP, CLOP and ROS were subjected to acidic, alkaline, oxidative, neutral and thermal degradation conditions. The degradation products obtained were well resolved from the pure drugs with significantly different Rf values. As the method could effectively separate the drugs from its degradation products, it can be used for stability-indicating analysis as well as statistical evaluation proved that the established method was accurate, specific, precise, repeatable and robust for the estimation of ASP, CLOP and ROS in bulk and pharmaceutical dosage form.

Keywords: High-performance thin-layer chromatography (HPTLC), Aspirin (ASP), Clopidogrel Bisulphate (CLOP) and Rosuvastatin Calcium (ROS), Stability indicating method development, Validation.


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