Validation Rp–Hplc Method for Simultaneous Estimation of Hydrochlorothiazide and Valsartan in Formulating Pharmaceutical form

Abstract

A novel, simple and accurate, Reversed Phase High Performance Liquid Chromatography (RP-HPLC) method for simultaneous estimation of Hydrochlorothiazide (HCT) and Valsartan (VAL) in mixture of standard and formulation tablets was validated in this research. The absorbance maximum of drugs using UV- spectroscopy was found at (318 and 250nm) for HCT and VAL respectively in deionized water: methanol mixture (60:40 V/V) as solvent. This method involves the separation of HCT and VAL on RP - HPLC Shimadzu type LC–20 - A, Japan, and Phenomenex C-18 (150 mm × 4.6 mm I.D) The elution was done using an eluent phase composed of 0.1 M ammonium acetate (AA), acetonitrile (ACN) and methanol (MOH) in the ratio of (50:25:25 V/V) with a pH adjusted at 3.0 using acetic acid). A separation was fixed for 10 min at 270 nm using a UV-Vis - detector and 1.0 mL/min flow rate. The drugs were eluted in (1.701 and 2.850 min.) for HCT and VAL respectively. The suitable conditions such as the elution phase composition, rate of flow, pH and wavelength were studied. The linearity of the method was in the range of concentration within (0.0125 – 100 and 0.1 – 100 μg/mL) for HCT and VAL respectively, while, R2 values within (0.9966 and 0.9963), and the means of recovery were found within (99.87 and 100.35) for HCT and VAL respectively. The method was applied for the estimation of gradient active of drugs in different formulating form samples. The method accuracy was validated by the mean of recovery percentages which, were found in acceptable limit.