Validated Hptlc Method For Simultaneous Determination Of Metformin Hydrochloride And Glibenclamide In Combined Dosage Form

Abstract

This paper describes a simple, accurate and precise HPTLC method for simultaneous determination of Metformin Hydrochloride and Glibenclamide in a bulk as well as in the combined dosages form. During optimization of Mobile phases different solvents viz. butanol, methanol and triethylamine were tried. However resolution between metformin hydrochloride and Glibenclamide was not proper. A mobile phase of Methanol: Water: 0.4 % sodium sulphate in water (7: 5:11) was developed for the densitometric determination of the drug products. Aluminum plate coated with the silica Gel 60 F254 was used as stationary phase. UV spectra were taken in the range of 190 nm to 400 nm. Densitometric evaluation of the separated bands of Metformin hydrochloride and Glibenclamide was performed at 232 nm and at 238 nm respectively. The Rf values of Metformin hydrochloride and Glibenclamide were 0.27 and 0.80 respectively. Developed method was Linear over the range of 250 ng to 1750 ng/spot and 250 ng to 1750 ng /spot of Metformin hydrochloride and Glibenclamide respectively. Precision of the method was evaluated by calculating RSD for peak response by inerday and intraday analysis. Results were Interday RSD of peak response=1.52 % and Intraday RSD of peak response=1.90 % for Metformin hydrochloride. Similarly for Glibenclamide interday RSD of peak response=1.67% and Intraday RSD of peak response = 0.92 %. Accuracy was determined in terms of percentage recovery at three concentration levels for Metformin hydrochloride and for Glibenclamide. The results for metformin hydrochloride at three concentration levels are 95.79%, 97.84 % and 96.06 %. In case of Glibenclamide the results are 96.0 %, 97.03 % and 96.53 %. Specificity was proved by spectral analysis of Metformin hydrochloride and Glibenclamide. There was no any interference of methanol being used as a diluents and mobile phase at the Rf values of Metformin hydrochloride and Glibenclamide. Standard spectrum and sample spectrum of Metformin hydrochloride and Glibenclamide were overlain to confirm Rf values. ICH