Abstract

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING HPLC METHOD FOR ESTIMATION OF RELATED SUBSTANCES IN IVABRADINE HYDROCHLORIDE IN SOLID ORAL DOSAGE FORM

Selva Kumar P* and Rajagopal K

ABSTRACT

Objective: A simple, accurate, linear, precise, specific, robust, rugged and stability indicating gradient high performance liquid chromatography (HPLC) method has been developed for estimation of related substances in Ivabradine hydrochloride from solid oral dosage form. Methods: The chromatographic separation was obtained using a mobile phase containing 20 mM ammonium acetate buffer pH 5.8 and methanol on Phenomenex Luna Phenyl-Hexyl column (250 × 4.6 mm, 5 μm), ambient temperature with UV detection at 285 nm at a flow rate of 1.0 mL/Minute. Results: The stability indicating capability of the method was proven by subjecting the drug to stress conditions as perICH recommended test conditions. The peak purity plots show that the Ivabradine hydrochloride peak is homogeneous and that there are no co eluting peaks indicating that the method is stability indicating and specific. The % RSD value was 0.81%, 1.44% and 2.19% for Impurity-B, Impurity-A and Impurity-3 respectively for method precision. Ruggedness of the method is indicated by the overall RSD values of 2.75%, 1.25% and 3.29% for Impurity-B, Impurity-A and Impurity-3 respectively. The linearity of response for Ivabradine hydrochloride and its related substances was determined with correlation coefficient of greater than 0.997. The LOQ for Ivabradine and the known impurities was at a level below 0.04%. The LOD for Ivabradine and the known impurities was at a level below 0.01%. The method has shown good, consistent recoveries for Impurity-B (90.99–107.70%), Impurity-A(98.77-106.22%) and Impurity-3 (94.01-104.36%). Conclusion: The analytical method validation data showed excellent results for precision, linearity, specificity, accuracy, ruggedness and robustness. The present method can be successfully used for routine quality control and stability studies.

Keywords: Force Degradation, Ivabradine hydrochloride, Related substances, RP- HPLC, Stability Indicating.