Development and Validation of a HPLC-UV Method for Antihypertensive Drug Amlodipine in Simulated Human Plasma Application to Protein Binding Studies

Abstract

A simple, sensitive, fast, and economical HPLC method was developed and validated for estimation of Amlodipine Antihypertensive Drug in simulated Human plasma. The validated HPLC method was used to quantify the concentration of selected actives in ultrafiltrate. Optimum separation conditions were obtained with Thermo β-basic C18 (100x 4.6 mm i.d., 5µ) column with mobile phase consisting of 10 mM Monobasic phosphate (pH 3.0) and acetonitrile (ACN) in gradient mode with column oven temperature maintained at around 25° C and elution monitored by a UV detector at 239nm. Protein precipitation was employed to extract the selected analyte form human plasma. The recoveries were more than 90%. The accuracy of samples for six replicate measurements was found within limit. The precision of samples for six replicate measurements was found within limit. The validated method was applied for quantitation of selected analytes in ultrafiltrate from protein binding experiments. The extraction efficiency of AMLO was found to be about 88.52%.