Two different botanical sources, Eclipta alba and Wedelia calendulacea are used as “Bhringaraja” in the Ayurvedic s ystem o f medicine. In the present study, an effort has been made to evaluate different sources by using high-performance thin-layer chromatography (HPTLC) as an analytical tool. Wedelolactone, one of the primary constituents of these plants, was taken as the marker compound for the evaluation. An HPTLC method was developed and validated for the evaluation of different sources of “Bhringaraja”. The chromatographic system was developed using silica gel 60 F254 HPTLC plates with the mobile phase toluene–chloroform–ethyl alcohol–formic acid (5:4:1:0.5, v/v). Linearity was found between the concentration ranges of 80 to 280 ng spot−1 (R2: 0.9994), and limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.36 ng spot−1 and 1.09 ng spot−1. The study has shown the presence of wedelolactone at the concentration of 0.26% w/w and 0.05% w/w in E. alba and W. calendulacea, respectively, whereas it was absent in another closely related species, Wedelia trilobata. At the same time, all the three plants were subjected to evaluation of quality-control parameters as per the World Health Organization (WHO) guidelines. Certain parameters such as foaming index, total alkaloids, and total bitter principles were significantly different in the three plants. Hence, the present HPTLC method development, and the validation and evaluation of quality-control parameters would be helpful in the standardization of individual plants and their formulations.
Authors:L. Sathiyanarayanan, A. R. Paradkar, and K. R. Mahadik
A sensitive, accurate, and robust high-performance thin-layer chromatographic (HPTLC) method has been established for simultaneous analysis of wedelolactone (WED) and asiaticoside (ASI) in Eclipta alba and Centella asiatica Linn., respectively. Chromatography was performed on silica gel with toluene-acetone-methanol-formic acid 3.0:2.0:2.0:0.05 (υ/υ) as mobile phase. Densitometric scanning at 317 nm for WED and at 530 nm, after derivatisation with 10% methanolic sulphuric acid, for ASI was used. The method was validated in accordance with the guidelines of the International Conference on Harmonization (ICH). RF values of 0.26 and 0.75 were obtained for ASI and WED, respectively. The linear ranges were 50–250 and 150–550 ng per band for WED and ASI, respectively, with good correlation coefficients (r2 = 0.999 and 0.9989, respectively). Accuracy was 99.29% and 99.45% for WED and ASI, respectively. The method was found to be precise, robust, and suitable for routine quality-control analysis of plant extracts and polyherbal formulations.