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determination of glimepiride in rat serum by rp-hplc method测定大鼠血清的glimepiride产物的方法.pdf

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determination of glimepiride in rat serum by rp-hplc method测定大鼠血清的glimepiride产物的方法

American Journal of Analytical Chemistry , 2011, 2, 152-157 doi:10.4236/ajac.2011.22017 Published Online May 2011 (http://www.SciRP.org/journal/ajac) Determination of Glimepiride in Rat Serum by RP-HPLC Method Sujatha Samala, Sandhya Rani Tatipamula, Ciddi Veeresham University College of Pharmaceutical Sciences, Kakatiya University, Warangal, India E-mail : ciddiveeresham@yahoo.co.in Received August 27, 2010; revised November 12, 2010; accepted November 17, 2010 Abstract A simple and sensitive reverse phase high performance liquid chromatography (RP-HPLC) method was de- veloped for the determination of glimepiride in rat serum. The assay involves one step liquid-liquid extrac- tion with methanol. Gliclazide was used as an internal standard. Chromatographic separation was performed on a C18 column using a mobile phase of methanol: 10 mM phosphate buffer (80:20 v/v) adjusted to pH 3.0 with orthophosphoric acid, at a flow rate of 1.0 ml/min and UV detection at 230 nm. The retention time of glimepiride and gliclazide was found to be 5.5 and 4.0 min and separation was complete in less than 10 min. The method was validated for linearity, accuracy and precision were found to be acceptable over the range of 0.5 - 500 µg/ml for glimepiride. The method was found suitable to analyse rat serum samples for application in pharmacokinetic, pharmacodynamic, bioavailability/bioequivalence studies. Keywords: Glimepiride, Gliclazide, RP-HPLC, Rat Serum 1. Introduction (UV) detection [6], high performance liquid chromatog- raphy (HPLC) with DAD [7] and UV detection [8] and The sulfonylurea glimepiride (1-[[p-[2-(3-ethyl-4-methy derivative UV spe