Résumé
Objective To investigate fingerprint analysis and quantification of Rhizoma Drynaria. Methods HPLC fingerprints were carried out at 30° on a Zirchrom C18 column (4.6 mm×200 mm, 5 µm). The mobile phase was methanol (A) and phosphoric acid aqueous solution (pH3.6, B). Gradient programmer was performed in linear gradient The chromatogram was monitored at a wavelength of 283 nm throughout the experiment and the chromatographic peaks were obtained, ranging from 200 nm to 400 nm. The injection volume was 10 µl. Then quantitative analyses were carried out at 30$ on a Kromasil C18 column (250 mm×4.6 mm,5 µm). Results In fingerprint analysis, plant materials from 16 regions were analyzed under the optimized HPLC conditions and 12 peaks were selected as characteristic peaks Compared with the reference standards, 6 major chromatographic peaks were characterized and identified, with sum of peaks area over 70% according to area normalization method. Additionally, the similarity analysis and HCA analysis were performed and the results got mutual authentication In quantitative analysis, the four compounds showed good regression (r>0.9995) within test ranges and the recovery of the method was in the range of 97.9%-103.7%. Conclusion The results revealed that the method of reference chromatographic fingerprints combined with multiple compounds determination could be used as an efficient strategy for systematic quality evaluation of Rhizoma Drynariae.
Résumé
Objective: To investigate the caffeic acid compounds from the roots of Arctium lappa and their neuroprotective activity. Methods: The compounds were isolated by column chromatography over silica gel, octadecylsilane (ODS) chemically bonded silica gel, Sephadex LH-20, and AB-8 macroporous resin coupled with preparative HPLC. Their structures were elucidated by spectroscopic analysis and their neuroprotective activity against glutamate-induced neurotoxicity was evaluated in SH-SY5Y cells by MTT assay. Results: Eight caffeic acid compounds were isolated from the roots of A. lappa, which were identified as 1,5-O-dicaffeoyl-3-O-(4-malic acid methyl ester)-quinicacid (1), 3,5-O-dicaffeoyl-quinic acid methyl ester (2), 3,4-O-dicaffeoyl-quinic acid methylester (3), 4,5-O-dicaffeoyl-quinic acid methyl ester (4), (2E)-1,4-dimethyl-2- [(4-hydroxyphenyl)methyl]-2-butenedioicacid (5), chlorogenic acid methyl ester (6), caffeic acid mtheyl ester (7), and 3,4,3',4'-tetrahydroxy-δ-truxinate (8). In vitro, these compounds showed the neuroprotective activity against glutamate-induced neurotoxicity on different levels. Conclusion: The neuroprotective activity of the roots in A. lappa against glutamate-induced neurotoxicity is related to the caffeic acid compounds. Compound 1 is a new compound; Compound 5 is a new natural compound; Compounds 2-4 and 6-7 are isolated from A. lappa for the first time; Compound 8 is isolated from the plants of Arctium L. for the first time.