DEVELOPMENT AND VALIDATION OF RP- HPLC METHOD FOR METFORMIN HYDROCHLORIDE AND NATEGLINIDE IN BULK AND COMBINED DOSAGE FORM
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
2. Patel JR, Suhagia BN, Patel BH. Simultaneous spectrophotometric estimation of Metformin and Repaglinide in a synthetic mixture. Indian J Pharm Sci 2007;69:844-6.
3. Keyur B, Emanual M, Patelia, Arpit S. Simultaneous estimation of Metformin hydrochloride and repaglinide in pharmaceutical formulation by HPTLC-densitometry method. J Chromatogr Sep Tech 2013;4:166-71.
4. Dalia Rashad W. Simultaneous determination of Metformin, Nateglinide and Gliclazide in pharmaceutical preparations using micellar liquid chromatography. Int J Biomed Sci 2012;8:144â€“51.
5. Mallikarjuna Rao N, Gowri Sankar D. RP-HPLC method for simultaneous estimation and stability indicating the study of Metformin and Linagliptin in pure and pharmaceutical dosage forms. Int J Pharm Pharm Sci 2015;7:191-7.
6. Deepali AN, Vidhya K, Sunil R. Dhaneshwar validated HPLC method for simultaneous quantitation of Benfotiamine and Metformin hydrochloride in bulk drug and formulation. Int J Pharm Pharm Sci 2013;5:138-42.
7. Ramanji Reddy T, Dhachinamoorthi D, Chandra Sekhar KB. Development of RPHPLC method for Metformin and Repaglinide in rabbit plasma. Int J Pharm Pharm Sci 2012;4:311-3.
8. Ediga Sasi K, Krishna Reddy V, Chandra K. A new simple RP-HPLC method for simultaneous estimation of Metformin hydrochloride and Gliclazide tablet dosage form. Int J Pharm Biol Sci 2012;2:277-83.
9. Vani R, Vijaya Kumer B, Krisshna Mohan G. Analytical method development and validation for the determination of Sitagliptin and Metformin using reverse phase HPLC method in bulk and tablet dosage form. World J Pharm Pharm Sci 2014;3:1803-11.
10. Sasi kiran E, Krishna reddy V, Chandra K. A new simple RPHPLC method for simultaneous estimation of Metformin hydrochloride and Gliclazide tablet dosage form. Int J Pharm Biol Sci 2012;2:277-83.
11. Deepa R, Laxmanbhai J, Madhabhai M. Stability indicating HPLC method for simultaneous determination of Repaglinide and Metformin hydrochloride in pharmaceutical dosage form. Int J ChemTech Res 2011;4:500-5.
12. International conference on harmonization of technical requirements for registration of pharmaceuticals for human use. Validation of analytical procedures; Text and methodology ICH Q2 (R1); 2005.