DEVELOPMENT AND VALIDATION OF RP- HPLC METHOD FOR METFORMIN HYDROCHLORIDE AND NATEGLINIDE IN BULK AND COMBINED DOSAGE FORM
Keywords:Keywords, Metformin hydrochloride, Nateglinide, RP-HPLC, Method development, Validation
Objective: To develop an accurate, precise and linear RP-HPLC method for simultaneous quantitative estimation of Metformin hydrochloride and Nateglinide in tablets and validate as per ICH guidelines.
Methods: The method used a reverse phase column, Inertsil C18-ODS 3V (250Ã—4.6 mm, 5 Âµm), a mobile phase comprising of phosphate buffer (pH 4.0): Acetonitrile: methanol (30:60:10) flow rate of 1.0 ml/min and a detection wavelength of 221 nm using a UV detector.
Results: The developed method resulted in elution of Metformin hydrochloride at 2.45 min and Nateglinide at 4.21 min. The calibration curves were linear (rÂ²=0.999) in the concentration range of 60-140 Âµg/ml and 14.4-33.2 Âµg/ml for Metformin hydrochloride and Nateglinide respectively. The percentage recoveries were found to be 99.59-101.36 for Metformin hydrochloride and 98.43-101.38 for Nateglinide. The LOD was found to be 2.18 Âµg/ml and 1.55 Âµg/ml for Metformin hydrochloride and Nateglinide respectively. LOQ was found to be 8.52Âµg/ml and 4.69Âµg/ml for Metformin hydrochloride and Nateglinide respectively.
Conclusion: A simple, accurate, precise, linear and rapid RP-HPLC method was developed for simultaneous quantitative estimation of Metformin hydrochloride and Nateglinide in bulk and pharmaceutical formulation and validated as per ICH guidelines. Hence, the method holds good for the routine analysis of Metformin hydrochloride and Nateglinide in various pharmaceutical industries as well as in academics.
Keywords: Metformin hydrochloride, Nateglinide, RP-HPLC, Method development, Validation
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