Stability Indicating RP-HPLC Method Development and Validation for the estimation of Clopidogrel bisulphate.

A simple, sensitive, accurate and specific stability-indicating high-performance liquid chromatographic method was developed and validated for the estimation of clopidogrel bisulphate in bulk. In the present study, extensive testing of clopidogrel bisulphate in different stress conditions were carried out as per the ICH guidelines Q1A (R2). The system consisted of a pump (Shimadzu, prominence, LC20AD), with 20µl sample injector, along with a PDA (Shimadzu, prominence, SPDM20A) detector at a wavelength of 254nm. Data was compiled using Shimadzu LC Solution software. The degraded products formed under various stress conditions were separated successfully from the drug by using a PHENOMENEX C8 Column (150 x 4.6mm, 5µm) with binary gradient conditions. Acetonitrile: phosphate buffer of pH 2.0 was used as mobile phase at flow rate of 1.2ml/min. Clopidogrel bisulphate was exposed to various stress conditions like oxidation, hydrolysis, photolysis and neutral decomposition. Clopidogrel bisulphate, which was found to degrade considerably in acidic, photo and oxidative conditions, was found to be stable in alkaline and neutral conditions. Apart from the formation of minor degraded products under accelerated conditions, the drug was reasonably stable in solid state. A good linear relationship over the concentration range of 150-500µg/ml was shown. Validation of the method was carried out as per the ICH guidelines. The method developed was found to be precise, accurate, specific and selective. Clopidogrel bisulphate showed degradation in 5M Hydrochloric acid at 80oC, in 3% hydrogen peroxide for 5min the drug showed around 35% of degradation, when exposed to sunlight for 15 min, formed around 25-30% of degradation products. Statistical analysis shows that the method is reproducible and selective for the estimation of clopidogrel bisulphate in dosage form.