High performance thin layer chromatographic method for the simultaneous quantification of glimepiride and 3-acetyl-11-keto-beta-boswellic acid along with robustness study using fractional factorial design

A simple, specific, and quantitative high-performance thin-layer chromatography method has been developed for the simultaneous determination of Glimepiride and 3-acetyl-11-keto-beta-boswellic acidin the formulation. Chromatographic development was performed using n-hexane: Ethyl acetate: Toluene: Glacial acetic acid (4:2:4:0.2, % v/v/v/v) as optimized mobile phase on TLC aluminum plate precoated withsilica gel 60 F₂₅₄ and further densitometric measurement at 242nm.The drugs were satisfactorily resolved with Rf values of 0.38±0.02, 0.65±0.02, for Glimepiride and 3-acetyl-11-keto-beta-boswellic acid, respectively. The linear ranges were found to be 100-600 and 4000-24000ng/band for Glimepiride and 3-acetyl-11-keto-beta-boswellic acid. The accuracy and precision measured were less than 2% relative standard deviation for both drugs. The sensitivity of the method in terms of limit of detection and limit of quantitation were found to be 11.22 and 34ng/band for Glimepiride and 439.92 and 1333.102ng/band for 3-acetyl-11-keto-beta-boswellic acid respectively. 2^4-1 fractional factorial design was applied for robustness study to evaluate the small difference in independent variables. Among four factors, the significant factor found was the volume of n-hexane that resulted in a change in the Rf of drugs. The proposed method was found to be accurate, precise, reproducible, robust, and specific.