Development and Validation of Bioanalytical Method for Determination of Telmisartan and Hydrochlorothiazide Using HPTLC in Human Plasma

Abstract

A simple, sensitive, rapid and economic high performance thin layer chromatographic method has been developed for determination of telmisartan and hydrochlorothiazide in human plasma using paracetamol as an internal standard. The plasma sample was extracted using mixture of methanol-acetonitrile (3.0:0.1, v/v). A concentration range from 200, 400, 600, 800, 1000, 1200 ng/spots were used for calibration curve of hydrochlorothiazide and telmisartan respectively. The percent recovery of telmisartan and hydrochlorothiazide was found to be75.98 and 81.91%. The mobile phase consists of chloroform: methanol: toluene (8:2:4 v/v/v). Densitometric analysis was carried out at wavelength 278 nm. The Rf values for hydrochlorothiazide, paracetamol and telmisartan were 0.28 ± 0.05, 0.50 ± 0.05, 0.66 ± 0.05 respectively. The stability of telmisartan and hydrochlorothiazide in plasma were confirmed during three freeze-thaw cycles (?20?C), on bench during 24 hours and post preparative during 48 hours. The proposed method was validated statistically and by performing recovery study for determination of telmisartan and hydrochlorothiazide in human plasma.