Analytical Method Development and Validation for Simultaneous Estimation of Metformin, Pioglitazone and Glimepiride in Combined Tablet Dosage Form by using RP-HPLC

M.D. Fiaz*, Dr. R. Vani, Dr. Iffath Rizwana
Post graduate student, Deccan School of Pharmacy, Hyderabad, Telangana, India.
University of Technology, Osmania University, Hyderabad, Telangana, India.

A B S T R A C T
Objective: To develop and validate a suitable method for the simultaneous estimation of Metformin, Pioglitazone and Glimepiride in Tablet dosage form by using RP-HPLC method. Methods: The chromatographic separation was performed on ZORBAX SB-PHENYL, 250X 4.6mm. The mobile phase was prepared by mixing Ammonium acetate and methanol in the ratio of (90:10%) v/v that run isocratically at the flow rate of 1.0ml/min. and detection of all the eluents is carried out by UV Detector. Results: The Rt of Metformin, Pioglitazone and Glimepiride were found to be 4.754 min., 6.390 min. and 9.948 min. respectively. Method was found to be linear over the range of 250-750 μg/ml for Metformin HCl, 7.5-22.5 μg/ml for Pioglitazone HCl and 0.5-1.5 μg/ml for Glimepiride. Percentage recoveries of Metformin, Pioglitazone and Glimepiride were obtained in the range of 101%, 100% and 99.66% respectively. The limit of detection of Metformin HCl, Pioglitazone HCl and Glimepiride is 2.203g/ml, 0.828g/ml and 0.007g/ml respectively. The limit of Quantification of Metformin HCl, Pioglitazone HCl and Glimepiride is 7.345g/ml, 0.2760g/ml and 0.024g/ml respectively. Conclusion: A new sensitive, simple, and stability indicating high performance layer chromatographic (HPLC) method has been developed and validated for determination of Metformin, Pioglitazone and Glimepiride. The proposed method can may be used for routine determination of Metformin, Pioglitazone and Glimepiride stability.
Keywords: Metformin, Pioglitazone and Glimepiride, RP-HPLC, UV Detector.