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Original Article
Year : 2018   |  Volume : 10  |  Issue : 1  |  Page : 41-47

Liquid chromatographic method for simultaneous estimation of Metformin HCl, Pioglitazone HCl and Glibenclamide in rat plasma

Separation of three analytes, namely metformin (MET), pioglitazone (PIO), and glibenclamide (GLB), was achieved within a single chromatographic run for the first time. Chromatographic method included stationary phase C18 Column (100 mm × 4.6 mm, 3.5 μm i.d.) and mobile phase consisting of ion-pair aqueous component and organic component in a gradient mode at 1 mL/min flow rate, and detection was monitored at 230 nm. Mobile-phase compositions and combinations were optimized for type and concentration of ion-pair reagents. Plasma sample preparation was based on protein precipitation by means of cold aqueous solution of 10% (w/v) trichloroacetic acid in combination with organic solvent addition. Ranitidine was used as the internal standard for MET, whereas rosiglitazone and amlodipine played the same role for PIO and GLB, respectively. Calibration curves were plotted from lower limit of quantification to 10,000 ng/mL for MET, PIO, and GLB. Weighing schemes of 1/X and 1/X2 were applied to observe goodness of fit in calibration curves. Precision was characterized by relative standard deviations below 15%. Stability analysis showed that all analytes are stable for at least 3 months when stored at −20°C. The validated method was applied for determination of MET, PIO, and GLB in pharmacokinetic study samples.
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