Development and Validation of RP-HPLC Method for Determination of Eprosartan Mesylate in Rat Plasma: Application to Preclinical Pharmacokinetic Study

Abstract

A selective and sensitive reverse phase high performance liquid chromatography (RP-HPLC) method was developed and validated for the determination of Eprosartan mesylate in rat plasma. A single step protein precipitation was carried out with acetonitrile to deproteinized plasma sample. A chromatographic separation was achieved on a Phenomenox, Gemini C18 (250x4.6 mm, 5 μm) column using acetonitrile and water (45:55) as a mobile phase with a pH adjusted to 3.4 with orthophsophoric acid (85%) at a flow rate 1 mL/min. The intensity of peak was monitored at 235 nm. The proposed chromatographic condition shows good symmetry and resolution of peaks. The retention time of Eprosartan mesylate and Olemesartan medoxamil (Internal standard) were appropriately 2.2 ± 0.5 and 3.1 ± 0.5 min, respectively. The validation studies performed as per ICH guidelines indicated high degree of accuracy, precision, with good degree of sensitivity and robustness of the proposed method. Furthermore, no interference was observed with plasma suggesting its utility for the pharmacokinetic analysis and bioavailability study of eprosartan mesylate in rat plasma. Key words : Eprosartan mesylate, Reverse phase high performance liquid chromatography, Pharmacokinetic analysis, Protein Precipitation, Validation, ICH guidelines.