Estimation of Metformin Hydrochloride, Gliclazide and Pioglitazone Hydrochloride: A UDDD-HPTLC Densitometry Method Validation Study

A simple and new UDDD-HPTLC method has been developed and validated for their estimation of MET, GLZ and PIO in bulk and combined tablet dosage form. Metformin (MET) is chemically, 1-carbamimidamido-N-N-dimethyl- methanimidamide. It is an oral anti-diabetic drug from the biguanide class. It is the first-line drug for the treatment of type-2 diabetes, particularly in overweight and obese people and those with normal kidney function and evidences suggest it may be the best choice for the people with heart failure.

The chromatographic separation of these drugs was carried out on precoated TLC plates silica gel 60F254by two mobile phases consisting of Ammonium Sulphate: Methanol: Acetonitrile: Water (4:3:2:1) for MET and PIO and Toluene: Ethyl Acetate: Formic Acid (6:4:0.5) for GLZ respectively for ideal separation and good resolution. The densitometric detection and quantification were carried out at 237 nm for MET and 200 nm for GLZ and PIO. The validation parameters were strictly followed as per the ICH guidelines.

Chromatographic separation of the standard solution of MET, GLZ and was performed. Briefly, the spot of the standard solution was applied on TLC plates. The TLC plates were developed by linear ascending development by using various solvents such as acetone, benzene, chloroform, ethyl acetate, methanol and toluene. The linearity range was obtained at 3000-8000ng/spot, 360-960 ng/spot, 90-240 ng/spot for MET, GLZ and PIO with r2value>0.999. The other parameters such as precision, reproducibility, robustness were efficiently obtained within the limits. The proposed method was successfully applied for simultaneous determination of MET, GLZ and PIO in the commercial formulation.

In simultaneous estimation, the different polarity of drugs makes it more cumbersome to develop and validate any chromatographic method. In comparison to HPLC, the suggested UDDD-HPTLC method is novel, less expensive, simpler, quicker, and more flexible. This technique can be used for routine quality control analysis because it enables simultaneous estimation of all API on a single TLC plate with a single application.