Development and Validation of a Novel High Performance Thin Layer Chromatography Method for Simultaneous Estimation of Mangiferin and Berberine in an Ayurvedic Medicine

Abstract

Setebid tablet is the natural antihyperglycemic agent with powerful antioxidant action. A highperformance thin layer chromatography method was developed for simultaneous estimation of mangiferin and berberine from setebid tablets. Camag Linomat 5 sample applicator was used for application of sample. Chromatographic separation of drugs was performed over thin layered chromatography plates pre-coated with silica gel 60F254 using toluene:acetone:formic acid (3.5:5.5:1 v/v/v) as the mobile phase via a linear ascending technique in twin trough chamber. Detection and quantification were carried out at an iso-absorptive wavelength of 261 nm using thin layered chromatography scanner III. This method showed good resolution for both drugs with retention factor 0.39±0.03 and 0.80±0.03 for mangiferin and berberine, respectively. The calibration curves were linear in the range of 400-800 and 100-500 ng/spot for mangiferin and berberine and correlation coefficients (r2) were 0.9995 and 0.9991, respectively. The method was validated according to the international conference on harmonisation Q2 R1 guidelines for accuracy, precision, limit of detection, linearity, limit of quantification, robustness and specificity. The limits of detection were 26.18 and 18.50 ng/spot for mangiferin and berberine, respectively. In conclusion, the developed method was rapid, simple, reliable and specific for the identification and quantification of mangiferin and berberine.