A NOVEL RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE QUANTIFICATION OF A POTENTIAL ANTI-DIABETIC DRUG METFORMIN HYDROCHLORIDE IN TABLET DOSAGE FORM

This study was conducted to develop a simple, economical, linear, rapid method for the assay studies of Metformin HCl by RP-HPLC method and to carry out the method validation.
Methods: A simple, robust and accurate method to carryout assay of Metformin hydrochloride tablet(500 mg) by RP-HPLC method in which the stationary phase used is Shimadzu shim-pack GIST C18 column with specification (5μm ×4.6×250 mm). This method involves isocratic elution of mobile
phase containing 70% buffer and 30% acetonitrile. The buffer used for analysis is Tetra-Butyl Ammonium Hydroxide (0.002%), the flow rate was maintained at 0.5 ml/min. detection was done at 232 nm. Principal peak for Metformin was observed at 3.5 min and the runtime for each injection was set to 10 min. The standard solutions of Metformin were prepared using purified water (milli-Q water) and scanned from 190 nm to 400 nm. Sharp peaks were observed in the range of 232 nm and thus, wavelength of 232 nm was selected and used throughout the validation process.This study was conducted to develop a simple, economical, linear, rapid method for the assay studies of Metformin HCl by RP-HPLC method and to carry out the method validation. Methods: A simple, robust and accurate method to carryout assay of Metformin hydrochloride tablet(500 mg) by RP-HPLC method in which the stationary phase used is Shimadzu shim-pack GIST C18 column with specification (5μm×4.6×250 mm). This method involves isocratic elution of mobile phase containing 70% buffer and 30% acetonitrile. The buffer used for analysis is Tetra-Butyl Ammonium Hydroxide (0.002%), the flow rate was maintained at 0.5 ml/min. detection was done at 232 nm. Principal peak for Metformin was observed at 3.5 min and the runtime for each injection was set to 10 min. The standard solutions of Metformin were prepared using purified water (milli-Q water) and scanned from 190 nm to 400 nm. Sharp peaks were observed in the range of 232 nm and thus, wavelength of 232 nm was selected and used throughout the validation process. Results: Linearity graph generated was found acceptable and accurate and the graph was generated in the range of 50% to 150% concentration. The regression coefficient was found to be 0.999(acceptable range). Validation was carried out according to ICH guidelines and found to be acceptable. Conclusion: This developed method was found to be simple, robust, economical, accurate, linear and can be used in the assay of Metformin tablet using RP-HPLC. Thanuja C, Ganesh M. RP-HPLC method for the simultaneous estimation of rosiglitazone and gliclazide in tablets. E-Journal of Chemistry. 2009;6(4):1188-92. 10.5530/phm.2014.1.5 Neelima K, Prasad YR. Analytical method development and validation of metformin, voglibose, glimepiride in bulk and combined tablet dosage form by gradient RP-HPLC. Pharm. 2014;5(1):27-33. doi: 10.5530/phm.2014.1.5. 10.4137/ACI.S31506 Bhole RP, Shinde SS, Chitlange SS, Wankhede SB. A high-performance thin layer chromatography (HPTLC) method for simultaneous determination of diphenhydramine hydrochloride and naproxen sodium in tablets. Anal Chem Insights. 2015;10:47-51. doi: 10.4137/ACI.S31506, PMID 26692760. 10.1186/s43094-021-00362-9 Musmade BD, Baraskar ML, Ghodke VN, Bhope SG, Padmanabhan S, Lohar KS. Impurity profiling method development and validation of metformin hydrochloride and teneligliptin hydrobromide hydrate in their combination tablet dosage form by using RP-HPLC with UV/PDA detector. Futur J Pharm Sci. 2021;7(1). doi: 10.1186/s43094-021-00362-9. 10.1016/j.jsps.2013.12.011 Chhetri HP, Thapa P, Van Schepdael A. Simple HPLC-UV method for the quantification of metformin in human plasma with one step protein precipitation. Saudi Pharm J. 2014;22(5):483-7. doi: 10.1016/j.jsps.2013.12.011, PMID 25473337. 10.1016/j.jsps.2013.12.011 Chhetri HP, Thapa P, Van Schepdael A. Simple HPLC-UV method for the quantification of metformin in human plasma with one step protein precipitation. Saudi Pharm J. 2014;22(5):483-7. doi: 10.1016/j.jsps.2013.12.011, PMID 25473337. 10.4103/0250-474X.51947 Mubeen G, Noor K. Spectrophotometric method for analysis of metformin hydrochloride. Indian J Pharm Sci. 2009;71(1):100-2. doi: 10.4103/0250-474X.51947, PMID 20177473. 10.4103/0250-474X.56022 Arayne MS, Sultana N, Zuberi MH, Siddiqui FA. Spectrophotometric quantitation of metformin in bulk drug and pharmaceutical formulations using the multivariate technique. Indian J Pharm Sci. 2009;71(3):331-5. doi: 10.4103/0250-474X.56022, PMID 20490308. 10.1002/bmc.3707 Chunduri RH, Dannana GS. Development and validation of a reliable and rapid LC-MS/MS method for simultaneous quantification of sacubitril and valsartan in rat plasma and its application to a pharmacokinetic study. Biomed Chromatogr. 2016;30(9):1467-75. doi: 10.1002/bmc.3707, PMID 26876742. 10.22159/ijcpr.2017v9i5.22151 PS, Vidyadhara S, Sasidhar RLC, Chakravarthi KK. Method development and validation for the estimation of entecavir in bulk and pharmaceutical dosage forms by RP-HPLC. Int J Curr Pharm Sci. 2017:107-11. doi: 10.22159/ijcpr.2017v9i5.22151.