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Activity-Guided Isolation and Identification of Radical Scavenging Components in Gao-Cha Tea

Lu, Rui-Li, Hu, Feng-Lin, Xia, Tao
Journal of food science 2010 v.75 no.8 pp. H239
inhibitory concentration 50, fractionation, quantitative analysis, laboratory techniques, medicinal properties, medicinal plants, herbal tea, chemical structure, free radical scavengers, mass spectrometry, Acer tataricum subsp. ginnala, antioxidant activity, thin layer chromatography
Gao-Cha is a traditional Chinese health tea made from Acer ginnala. We performed a components and radical scavenging activity analysis to identify any medicinal components in this tea. High performance thin layer chromatography (HPTLC)-1,1-Diphenyl-2-picryhydrazyl (HPTLC-DPPH) assay showed that the methanolic extract contained strong radical scavengers. Quantative analysis revealed that the IC₅₀ of the extract against 1 mM DPPH was 52.7 ± 0.6 μg/mL. Bioactive-guided isolations led to procurement of 3 radical scavengers with IC₅₀s of 17.5 ± 2.1, 29.3 ± 2.5, and 21.6 ± 1.7 μg/mL, respectively. Analysis of the high resolution-electric spray ionization-mass spectrometer and ¹H, ¹³C, distortionless enhancement by polarization transfer at 135°, heteronuclear quantum coherence, correlating spectroscopy coupling, and heteronuclear multiple bond coherence (HMBC) data revealed that the compounds were methyl 3, 4, 5-trihydroxybenzoate (1), quercetin-3-O-α-rhamnopyranoside (2), and 2,6-bis (3,4,5-trihydroxybenzoyl)-aceritol (3). Bioactive combined components analysis revealed that, apart from compounds 1, 2, and 3, the tea possibly contained radical scavengers: ginnalin A (4) and B (5), 2″-O-Galloylquercitrin (6) and 3″-O-Galloyl-quercitrin (7). Compounds 2, 6, and 7 were isolated from Acer ginnala for the first time. The positions of the 2 galloyl moieties in compound 3 were unambiguously established by the HMBC spectrum for the first time.