J Nanomedic Nanotechnol ISSN:2157-7439 JNMNT an open access journal Journal of Nanomedicine & Nanotechnology - Open Access Research Article OPEN ACCESS Freely available online doi:10.4172/2157-7439.1000102 Volume 1• Issue 1•1000102 Estimation of Metformin in Bulk Drug and in Formulation by HPTLC Shweta Havele and Sunil Dhaneshwar* Research and Development Centre in Pharmaceutical Sciences and Applied Chemistry, Poona College of Pharmacy, Bharati Vidyapeeth University, Erandwane, Pune – 411038, India Keywords: Thin layer chromatography; Pharmaceutical analysis; Antidiabetic drug; Metformin; Tablet; Bulk drug Introduction Metformin (Metformin HCl, N,N-dimethylimidodicarbonimidic diamide hydrochloride, MET, Figure 1) [1] is an oral antidiabetic drug [2]. Metformin hydrochloride is formulated as tablet dosage forms. This drug was approved by the FDA in December 1994 and has been the only clinically available drug that can significantly improve insulin sensitivity in patients that suffer from Diabetes type II (non-insulin dependent). Typically Metformin reduces basal and postprandial hyperglycemia by about 20.5% in more than 90% of the patients. Determination of MET in plasma by various analytical methods like HPLC MS or UV detector [3-11], in formulation [12] multicomponent system, HPLC [13,14]. Several other HPLC methods were also developed for the determination of metformin hydrochloride. But these methods are sophisticated, expensive and time consuming as compared to simple HPTLC method. There is a need for a simple, rapid, cost effective and reproducible method for assay of MET in its dosage forms. Therefore, it was thought of interest to develop simple, rapid, accurate, specific and precise HPTLC method for the analysis of MET in its tablet formulation. The objective of the current work is, therefore, to develop a simple HPTLC method for analysis of metformin hydrochloride in tablet formulations. Experimental Materials MET working standard was a generous gift from Ranbaxy, Indore, India. Silica gel 60 F 254 TLC plates (10 × 10 cm, layer thickness 0.2 mm, E. Merck, Darmstadt, Germany) were used as a stationary phase. All chemicals and reagents used were of analytical grade and were purchased from Merck Chemicals, India. Glycomet containing 500 mg of MET were purchased from USV limited, Biciphage containing 850 mg MET were purchased from Biochem and Bigomet containing 250 mg of MET were purchased from Genetica (Aristo). Instrumentation The HPTLC system consisted of a Camag Linomat 5 semi-automatic spotting device (Camag, Muttenz, Switzerland), a Camag twin-trough chamber (10 cm × 10 cm), Camag winCATS software 1.4.4.6337 and a 100 l Hamilton syringe. Sample application was done on precoated silica gel 60 F 254 TLC plates (10 cm × 10 cm). TLC plates were pre- washed with methanol and activated at 80°C for 5 min prior to the sample application. Densitometric analysis was carried out utilizing Camag TLC scanner 3. Preparation of standard solutions A stock solution of MET was prepared by dissolving 100 mg in 100 ml methanol (1000 g/ml). Further standard solutions were prepared by dilution of the stock solution with methanol to reach a concentration range 200-1000 ng/spot. Sample preparation Three brands of tablets Glycomet of (USV limited), Bigomet (Genetica (Aristo)) and Biciphage of (Biochem Pharmaceuticals) were selected. Twenty tablets were weighed and the average weight was calculated. The tablets were then powdered and an amount equivalent to one tablet was dissolved in 50 ml methanol. To ensure complete extraction of the drug it was sonicated for 45 min. This solution was filtered through a Whatman no. 41 paper. *Corresponding author: Sunil Dhaneshwar, Research and Development Centre in Pharmaceutical Sciences and Applied Chemistry, Poona College of Pharmacy, Bharati Vidyapeeth University, Erandwane, Pune – 411038, India, E-mail: sunil. dhaneshwar@gmail.com Received October 02, 2010; Accepted October 25, 2010; Published October 27, 2010 Citation: Havele S, Dhaneshwar S (2010) Estimation of Metformin in Bulk Drug and in Formulation by HPTLC. J Nanomedic Nanotechnolo 1: 102. doi:10.4172/2157- 7439.1000102 Copyright: © 2010 Havele S, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract A simple and sensitive, HPTLC method has been developed for the quantitative estimation of metformin in its single component tablet formulation. Metformin was chromatographed on silica Gel 60 F 254 TLC plate using ammonium sulfate (0.5%): 2-propanol: methanol in the ratio of 8.0:1.6:1.6 (v/v/v) as mobile phase. Metformin showed R f value of 0.50±0.03 was scanned at 238 nm using Camag TLC Scanner 3. The linear regression data for the calibration plot showed a good relationship with r =0.999. The method was validated for precision and recovery. The limits of detection and quantification were 95 and 200 ng/spot respectively. The developed method was successfully used for the assay of metformin tablet formulations. The method is simple, sensitive and precise; it can be used for the routine quality control testing of marketed formulations. Figure 1: Chemical structure of Metformin. NH NH N H N NH 2 H 3 C CH 3