Development and Validation of an Assay for Bisoprolol Fumarate and Hydrochlorothiazide Multicomponent Drug

Abstract

A simple, precise, accurate and economical Reverse phase HPLC with a detection UV, method was developed and validated for the simultaneous determination of Bisoprolol fumarate and Hydrochlorothiazide in pharmaceutical formulations. Better separation was achieved using Cyanide column (250mm × 4.6mm, 5μm), maintained at 30 ͦ C. The mobile phase was composed of buffer: methanol in the ratio of 82:18 (v/v), (the buffer was 1% solution of tetra butyl ammonium hydroxide, adjusted to pH = 5.0 by acetic acid). Isocratic elution was used with a flow rate of 0.9 ml/min, injection volume was 10μ, and effluents were monitored by UV (228nm). The retention time of Bisoprolol fumarate and Hydrochlorothiazide was 4.7min and 7.7min, respectively. The method was tested for linearity over a concentration range of (40-160) μg/ml and (100-400) μg/ml, for Bisoprolol fumarate and hydrochlorothiazide, respectively; the correlation coefficient (R2), was found to be 0.9998 and 0.9999, respectively. The method successfully passed all validation tests stipulated in the validation protocol of International Conference on Harmonization (ICH) and united State Pharmacopeia (USP).