Validated stability-indicating high performance thin layer chromatographic method for determination of Ivabradine hydrochloride in bulk and marketed formulation: An application to kinetic study

Abstract

A sensitive, selective, precise and accurate stability-indicating high-performance thin layer chromatographic method for analysis of Ivabradine hydrochloride (IH) an anti anginal agent, both as a bulk drug and in formulations was developed and validated according to ICH guideline. Densitometric analysis of IH was carried out in the absorbance mode at 287nm using ethyl acetate: 0.389M ammonium acetate in methanol (1:5, v/v) as solvent system. This system was found to give compact spots for IH at an Rf value of 0.36±0.01. Moreover, IH was subjected to acid and alkali hydrolysis, oxidation, accelerated humidity/temperature, wet heat treatment, and photo degradation. The drug undergoes degradation under mainly acidic and basic conditions. Also the degraded products were well resolved from the pure drug with significantly different Rf values. Linearity was found to be in the range of 1200–2800ng/band. The LOD and LOQ for IH were 255.86ng/band and 775.33ng/band, respectively. “Bartlett’s test” and “Lack of fit” applied on peak area for linearity, additionally proved validity of the developed method. Good accuracy and precision were obtained as revealed from %RSD value less than 2. Similarly, no interference was observed from common excipients in tablet formulation as well as degradation product, indicating specificity of the method. As the method could effectively separate the drug from its degradation product, it can be employed as a stability-indicating one. Moreover, proposed method was also utilized to investigate the kinetics of acidic degradation process at different temperatures and first order rate constant, half-life, shelf-life and activation energy were calculated.