DEVELOPMENT AND VALIDATION OF STABILITY INDICATING HPTLC METHOD FOR ESTIMATION OF GLIMEPIRIDE AND METFORMIN HYDROCHLORIDE

An Accurate, sensitive, precise and stability indicating high performance thin layer chromatographic method has been developed and validated for the estimation of Glimepiride and Metformin hydrochloride. The method was developed using TLC aluminium plates precoated with silica gel 60F₂₅₄ as the stationary phase and 0.5% Ammonium Sulfate: Methanol (7.5:2.5 v/v) as mobile phase. Densitometric analysis of Glimepiride and Metformin hydrochloride was carried out in the absorbance mode at 228 nm. The two drugs were satisfactorily resolved with R_F values 0.73 and 0.45 for Glimepiride and Metformin hydrochloride respectively.  Linearity was found to be 600-2100 ng/band for Glimepiride and 200-700 ng/band for Metformin hydrochloride. The method was found to be accurate, precise, and robust according to acceptance criteria. The limit of detection (LOD) was found to be 0.05 ng/band and 0.32 ng/band for Glimepiride and Metformin hydrochloride respectively. The limit of quantification (LOQ) was found to be 0.16 ng/band and 0.96 ng/band for Glimepiride and Metformin hydrochloride respectively. The proposed HPTLC method was successfully applied for the forced degradation study of Glimepiride and Metformin hydrochloride. Forced degradation study was carried out for both the drugs in oxidative condition 0.3% H2O2 for 30mins, in acidic condition 0.1 M HCl at 80˚C for 60min, in basic condition 0.01 M NaOH at 80˚C for 30 min, in thermal condition 60˚C for 24 hours and in photolytic condition for 24 hours. Degradation products were well separated by proposed method