Phytoestrogenic activity of Aceriphyllum rossii and rapid identification of phytoestrogens by LC-NMR/MS and bioassay-guided isolation

Aceriphyllum rossii Engler (Saxifragaceae) has been used as a nutritious food in Korea, but only minimal studies on this plant have been undertaken. We have examined the estrogenic effect of A. rossii and have discovered its potent constituents. To identify the estrogenic activity of A. rossii, we measured the transactivation of the estrogen-responsive element (ERE) in MCF-7 cells transfected with an ERE-containing construct after treatment of the extract and fractions of A. rossii. A. rossii showed a potent estrogenic activity and gallic acid ethyl ester; quercetin and kaempferol were identified as major constituents of the active fractions by liquid chromatography-nuclear magnetic resonance spectroscopy/mass spectrometry. We confirmed quercetin and kaempferol are the major active constituents after activity-guided isolation of seven compounds: gallic acid ethyl ester (1), quercetin (2), kaempferol (3), quercetin-3-O-(6aEuro(3)-galloyl)-beta-d-glucopyranoside (4), quercetin-3-O-beta-d-glucopyranoside (5), kaempferol-3-O-(6aEuro(3)-galloyl)-beta-d-glucopyranoside (6), and kaempferol-3-O-beta-d-glucopyranoside (7). These results show A. rossii could be a possible candidate for the treatment of postmenopausal symptoms.