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Development of HPTLC Method for Determination of Amlodipine Besylate and Olmesaratan Medoxomil Using Human Plasma by Liquid Liquid Extraction

Ambadas. R Rote and Sadhana.K. Kande

A simple, sensitive, rapid and economic high performance thin layer chromatographic method has been developed for determination of amlodipine besylate and olmesaratan medoxomil in human plasma by liquid liquid extraction using paracetamol as an internal standard. The plasma sample was extracted using mixture of methanol: acetonitrile (3.0: 0.1). A concentration range from 500-3000 ng/spot of amlodipine besylate and olmesaratan medoxomil was used for calibration curve respectively. The percent recovery of amlodipine besylate and olmesaratan medoxomil was found to be 90.41 and 90.64 % respectively. The mobile phase consists of chloroform: methanol (9:1, v/v). Densitometric analysis was carried out at wavelength 254 nm. The Rf values for amlodipine besylate, olmesaratan medoxomil and paracetamol was 0.50 ± 0.05, 0.26 ± 0.05 and 0.66 ± 0.05 respectively. The stability of amlodipine besylate and olmesaratan medoxomil in plasma was confirmed during three freeze-thaw cycles at ‘-20°C’, on bench during 24 h and post preparative during 48 h. The proposed method was validated statistically and by performing recovery study for determination of amlodipine besylate and olmesaratan medoxomil in human plasma by liquid liquid extraction.