Abstract

DEVELOPMENT AND VALIDATION OF A HPTLC METHOD FOR SIMULTANEOUS ESTIMATION OF KAEMPFEROL AND GLYCYRRHIZIC ACID

Vishakha R. Niman*, Varsha M. Jadhav and Vilas Kadam

ABSTRACT

A simple, sensitive, precise, specific, and accurate high-performance thin layer chromatography (HPTLC) method was developed for simultaneous estimation of Kaempferol and Glycyrrhizic acid from the extract of Nyctanthes arbor-tristis and Glycyrrhiza glabra. Chromatographic separation of drugs was performed over TLC plates pre-coated with silica gel 60F254 using toluene: acetone: formic acid (4.5:4.5:1 v/v/v) as the mobile phase via a linear ascending technique. Detection and quantification were carried out at an iso- absorptive wavelength of 264 nm. This method showed good resolution for both drugs with retention factor 0.29±0.03 and 0.75±0.03 for Kaempferol and Glycyrrhizic acid, respectively. The calibration curves were linear in the range of 100-700 and 100-700 ng/spot for Kaempferol and Glycyrrhizic acid and correlation coefficients (r2) were 0.9926 and 0.9942, respectively. The method was validated according to the International Conference on Harmonisation (ICH) Q2 R1 guidelines for accuracy, precision, and the limit of detection, linearity, the limit of quantification, robustness, and specificity. The limits of detection were 32.19 and 29.52 ng/spot for Kaempferol and Glycyrrhizic acid, respectively. In conclusion, the developed method was rapid, simple, reliable, and specific for the identification and quantification of Kaempferol and Glycyrrhizic acid.

Keywords: High-performance thin layered chromatography, Simultaneous estimation, Kaempferol and Glycyrrhizic acid.