Method development and validation of simultaneous estimation of metformin and glibenclamide in combined tablet dosage form by RP -HPLC method

Abstract

A new simple, accurate, rapid and precise isocratic high performance liquid chromatographic (HPLC) method was developed and validated for the determination of Metformin and Glibenclamide in tablet formulation. The proposed HPLC method utilizes Develosil ODS HG-5 RP C18, 250 mm x 4.6 mm I.D; 5 μm with a flow rate of 1.0 mL/min, mobile phase consisting of Methanol : Acetate buffer (pH=3.0) = 75:25 (v/v) at a detection wavelength 256 nm. The method was validated in terms of accuracy, precision, linearity, limits of detection, limits of quantitation, and robustness. This optimized method has been successively applied to pharmaceutical formulation and no interference from the tablet excipients was found.  Retention times of Metformin and Glibenclamide were found to be 2.24 min, and 3.28 min with a tailing factor 1.30, 1.29and 2603, 3534 as theoretical plates respectively which are within the limits. All the parameters were validated according to the ICH guidelines and found to be within limits. Limit of detection (LOD) and Limit of quantification (LOQ) were estimated from the signal-to-noise ratio. The LOD values of Metformin and Glibenclamide were found to be 0.062 and 0.018 µg/mL respectively. Metformin and Glibenclamide LOQ’s were found to be 0.19, and 0.056µg/mL respectively. Linearity ranges for Metformin and Glibenclamide were 2-10 µg/mL, and 3-15 µg/mL respectively. Percent recovery study values of Metformin and Glibenclamide were found to be within 98-100 %. This new method was successfully developed and validated as per ICH guidelines, can be utilized for the quantitative estimation of Metformin and Glibenclamide in pharmaceutical dosage forms.