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DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR THE DETERMINATION OF ROSIGLITAZONE IN HUMAN PLASMA
Ahmed Yusuf, Syed N. Alvi and Muhammad M. Hammami*
ABSTRACT A simple reversed-phase high performance liquid chromatographic (RP-HPLC) assay for the determination of rosiglitazone level in human plasma was developed and validated. After acidification with 1 M HCL, rosiglitazone and glibencamide (internal standard, IS) were extracted with a mixture of methylene chloride and hexane (50:50, v:v), efficiently separated at room temperature on a Nova-pak C18 cartridge with retention times of 6.4 and 5.2 minutes, respectively, and detected with multi-wave length fluorescence detector. The mobile phase consisted of 0.01 M dibasic potassium hydrogen phosphate (pH 6.5) and acetonitrile (65:35, v:v) and was delivered at a flow rate of 2.0 ml/min. No interference in blank plasma or by commonly used drugs was observed; and the detection limit of rosiglitazone in plasma was 2.5 ng/ml. Calibration curves were linear (R2 ≥ 0.9988) over the range of 5–800 ng/ml, and inter-day coefficient of variation (CV) was ≤ 7.7%. Mean extraction recovery of rosiglitazone and the IS was ≥ 91%, whereas inter-day bias of rosiglitazone measurement was (-3.2% to 5.3%). The method was used to assess the stability of rosiglitazone in human plasma under various conditions in the clinical laboratory. Further, it was successfully employed to measure rosiglitazone level in samples obtained from a healthy volunteer. Keywords: Rosiglitazone, Glibencamide, Human plasma, HPLC. [Download Article] [Download Certifiate] |