Development and validation of an HPTLC method for estimation of coronarin D in hedychium coronarium rhizome

Abstract

Rhizome extracts of Hedychium coronarium are widely used as phytotherapeutics. As of date, there is no documented study on the standardization of H. coronarium extract, and the following research is an effort in this direction. Coronarin D is an important bioactive compound present in H. coronarium which shows chemopreventive activity against cancer. H. coronarium extracts were assessed for coronarin D content for the first time. The extraction was checked using different solvents: n-hexane, acetone, and methanol. Coronarin D was separated on silica gel 60F254 high-performance thin-layer chromatography (HPTLC) plates by isocratic gradient method using n-hexane-ethyl acetate (80:20 v/v) as mobile phase. Densitometric quantification was performed at 231 nm in absorption mode. This method gave a well-defined peak at Rf 0.20 corresponding to coronarin D. The method was validated using International Conference on Harmonization (ICH) guidelines in terms of precision, repeatability, and accuracy. Linearity range of coronarin D was 200-1000 ng spot-1 with a correlation coefficient of R2 ± SD = 0.9987 ± 2.62% in the concentration range of 200-1000 ng spot-1. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 35 and 115 ng, respectively. Accuracy of the method was checked by recovery studies conducted at three different concentration levels, and the average percentage recovery was found to be 98.22 % for coronarin D. Among the different solvents, acetone produced maximum extraction efficiency of coronarin D. The proposed HPTLC method can be applied for robust identification and quantitative determination of coronarin D in H. coronarium extracts.