Liquid Chromatographic–Mass Spectrometric Determination of Ezetimibe in Human Plasma With Validation Approach

Context: Ezetimibe (EZ) is a hydroxymethylglutaryl CoA reductase inhibitor, which is classified as an anti-hyperlipidemic drug. Aim: The current research aims to describe a high throughput double liquid extraction technique to extract EZ from human plasma. Materials and Methods: The quantification of free and total EZ in human plasma was achieved by the creation of a straightforward, quick, and highly sensitive mass detection method by applying the liquid chromatographic technique. Using methyl tertiary-butyl ether and tolterodine tartrate as an internal standard, a double liquid extraction approach was adopted to take the analytes out from samples of human plasma. Results: Reconstituted samples were then chromatographed using 80:20, v/v combination of acetonitrile and ammonium acetate 5 mM as the mobile phase at a flow rate of 0.5 mL/min on a reversed-phase column (C18, 25 mm × 4.6 mm, 5 µm from Waters). For both free and total EZ, standardization curves for the concentration range were linear (r2 > 0.99) through a range of 0.1–15.0 and 1.0–150.0 ng/mL, respectively. Precision and accuracy procedures conducted inside and between days yielded results that fell within acceptable bounds. Conclusion: Analyzing a large number of plasma samples per day was made feasible by a 0.5-min run time for each sample. Human plasma samples were successfully analyzed using the current method, which can be used as a quick, accurate, and appropriate for further regular sample assessment in pharmacokinetic research. Key Messages: Our study has determined successfully ezetimibe both in total and free forms in human plasma by applying the validated liquid chromatography–mass spectrometric method.