Simultaneous determination of chlorogenic and caffeic acid in Siegesbeckia orientalis L. (Xi Xian) by a validated high-performance thin-layer chromatographic method

Abstract

Secondary metabolites are an integral part of the plant kingdom which plays a vital role in development, growth, and defense. These are also very helpful for humans against different disorders. Phenols due to their strong antioxidant activity have a central role in conferring different types of biological activities. The present study aims at the simultaneous determination of chlorogenic acid and caffeic acid in the methanolic extracts of leaf samples of Siegesbeckia orientalis L. Simultaneous elution of both compounds has been attained by using toluene-ethyl acetate-formic acid-methanol (6:6:1.6:0.6 v/v) as the mobile phase, and densitometric evaluation was done at 366 nm. The linear regression data for the calibration curve of both compounds show good linear relationship in the concentration range of 200-600 ng. The proposed method has been validated in terms of limit of detection, limit of quantification, specificity, precision, and accuracy studies. The present high-performance thin-layer chromatographic method has been applied for the quantification of both compounds in the various accessions of S. orientalis collected from different localities of North India. Maximum amount of chlorogenic acid was found in the Taluka accession, while caffeic acid was found in the Rarhi accession collected from Uttarakhand state.