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Differentiation of Cirsium japonicum and C. setosum by TLC and HPLC‐MS
Author(s) -
Ganzera Markus,
Pöcher Astrid,
Stuppner Hermann
Publication year - 2005
Publication title -
phytochemical analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.574
H-Index - 72
eISSN - 1099-1565
pISSN - 0958-0344
DOI - 10.1002/pca.846
Subject(s) - chemistry , chromatography , high performance liquid chromatography , luteolin , flavonoid , acetic acid , biochemistry , antioxidant
The Chinese Pharmacopoeia indicates the use of field thistle ( Cirsium setosum ) and Japanese field thistle ( C. japonicum ) in the treatment of bleeding and inflammation. In the absence of an analytical method for the differentiation and analysis of these two species, TLC and HPLC‐MS methods have been developed for this purpose. Both species could be readily distinguished by their flavonoid pattern as revealed by TLC on silica gel layers eluted with ethyl acetate:formic acid:acetic acid:water. The quantitative determination of four flavonoids, namely hispidulin‐7‐neohesperidoside, linarin, pectolinarin and luteolin, was possible using HPLC. Their optimum separation was achieved on a C 12 column eluted with water and 0.025% trifluoroacetic acid in acetonitrile. HPLC‐MS experiments were performed to confirm peak identity. In samples of C. japonicum , pectolinarin was the major flavonoid (0.32–2.00%), followed by linarin, hispidulin‐7‐neohesperidoside and luteolin; the total flavonoid content varied from 0.81 to 3.67%. In C. setosum only one flavonoid (linarin; 1.36–2.83%) was assignable. The HPLC method was validated for linearity, limit of detection (≤1.7 ng on‐column), peak purity, repeatability (≤2.3%) and accuracy (recovery rates of spiked samples were between 99.2 and 101.6%). Copyright © 2005 John Wiley & Sons, Ltd.