High performance liquid chromatographic method for determination of ezetimibe in pharmaceutical formulation tablets

Ezetimibe (Fig. 1) is an anti hyperlipidemic and is usually categorized as HMG CoA reductase inhibitor (1-2). Ezetimibe belongs to a class of lipid lowering compounds that selectively inhibits intestinal absorption of cholesterol and related phytosterols .It potentially inhibits the transport of cholesterol across the intestinal walls there by reducing plasma cholesterol (3). Ezetimibe coadministered with HMG CoA reductase inhibitors is licensed for the treatment of primary hypercholesterolemia in patients, and for homozygous familial hypercholesterolemia (4). Various methods have been reported for estimation of ezetimibe in pharmaceutical formulations which includes the muse of spectrophotometry (5‐12), Capillary Zone Electrophoresis (13), HPLC (14‐17), and LC -MS (1819) methods. Eranda was analysed the ezetimibe by HPLC but theirs method have a long time for run time for HPLC (16). Pawer applied the HPLC method but this method was a low recovery of ezetimibe(5). Sistla used the HPLC method for analysis of ezetimibe but this method was not high accuracy (14). Singh used for analysis of ezetimibe from HPLC method but this method was not simple and no precision (15). Although these methods were sufficiently sensitive, they were not suitable for most laboratories to perform studies involving samples in high through-put for therapeutic monitoring. The problems of these methods are, the long analysis time, large volume of sample, or low extraction recovery may not meet the requirement for high throughput, speed and sensitivity in bio sample analysis for quantitative analysis. As a result, a simple method that can determine ezetimibe in tablets formulation was required. Present studies involves Abstract Ezetimide belongs to a class of lipid lowering compounds that selectively inhibits intestinal absorption of cholesterol and related phytosterols. The purpose of this study is to establish a reliable and quick method for the assignment of ezetimibe in tablets form by high performance liquid chromatography with ultraviolet detection (HPLC-UV). A rapid and sensitive HPLC method has been developed for determination of ezetimibe in tablets formulation. Mobile phase was composed of acetonitrile-ammonium acetate (10 mM, pH 3.0), 75:25 (v/v) with a flow rate of 1 ml/min. The eluted peaks were detected by a UV detector was set at wavelength of 240 nm. The method results in excellent separation with good resolution of analyte. Standard curves were linear (r = 0.996) over the wide ezetimibe concentration range of 10-60.0 μg mL with acceptable accuracy and precision. The limits of detection (LOD) and quantitation (LOQ) of the method were 5 and 10 μg/ml, respectively. The average drug recovery was 95.3% throughout the linear concentration range. Statistical assessment of various in vitro dissolution parameters and assay results was also conducted to establish if there were any significant difference among them. The validated HPLC method has been used successfully to study ezetimibe. Due to simplicity, rapidity and accuracy of the method, we believe that the method will be useful for routine quality control analysis.