Quantitative bioanalytical and analytical methods for estimation of ivabradine hydrochloride in pure and pharmaceutical dosage form

Abstract

Introduction: New analytical and bioanalytical methods were developed for the estimation of Ivabradine hydrochloride in bulk and pharmaceutical dosage form by UV spectrophotometry and high-performance liquid chromatography (HPLC) technique. Objective: The primary objective of the study is to develop a new RP-HPLC method for estimation of Ivabradine Hydrochloride in pure and formulation and to develop a bioanalytical method for analysis of Ivabradine Hydrochloride in biological samples. The methods were validated as per International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use and USFDA guidelines respectively. Methods: In Reversed-phase (RP)-HPLC method developed telmisartan is the internal standard used. After liquid–liquid extraction, the analyte and the internal standard were chromatographed on Waters 125A (10 µ, 300 × 3.9 mm) C18 column using 20-µL injection volume with a run time of 15 min. An isocratic mobile phase consisting of acetonitrile and ammonium acetate buffer pH 7.8 (60:40% vol/vol) is used to separate drug and internal standard. In Spectrophotometric bioanalytical method developed for the estimation of ivabradine hydrochloride in pure and pharmaceutical dosage form. The solvent system used is absolute methanol and detected at the wavelength of 287 nm. The solvent system used is absolute methanol and detected at the wavelength of 287 nm. Results: The Analytical method is validated according to ICH guidelines over the range of 2–16 μg/ mL, showing accuracy, precision, selectivity, and robustness. For Bioanalytical method the linearity is established in the range of 500–3500 ng/mL with the regression coefficient of r2 = 0.9994. The validated spectrophotometric method is used successfully to study ivabradine hydrochloride in rat plasma and also quantitative determination in marketed tablets. Conclusion: The proposed methods were successfully applied for the quantitation of ivabradine hydrochloride in pharmaceutical dosage form with good recovery and reproducibility.