Curculigo orchiodes GAERTN. is a tiny herbal plant widely distributed in China, India, Malaya, Japan and Australia. Its rhizome, known as "Xianmao" in China, is a common kidney-Yang-reinforcing and anti-aging traditional Chinese medicine. It has the properties of warming kidney, invigorating Yang, expelling cold and eliminating dampness, and is used to treat impotence, enuresis, cold sperm, cold pain of back and knee, and numbness of the limb. 1,2) In India, the tuberous root of this plant is also considered to be tonic, alterative, demulcent, diuretic, and restorative, and is used as a poultice for itch and skin disease.3) Previous phytochemical investigations on the rhizomes of this species revealed the presence of curculigoside, 4,5) curculigoside B, 5,6) curculigoside C, 5) orcinol glycosides, 4) 2,6-dimethoxyl benzoic acid, curculigines A-C, 7) curculigol, 8) curculigosaponins A-M, 9) 1,3,7-trimethylxanthine, daucosterol, 4) and aliphatic longchain ketones. 10) Apart from curculigoside, which showed high phagocytic activity, 4) the bioactivities of these compounds from this plant were not clear. Venukumar and Latha reported that the methanol extract of C. orchioides served as an effective hepatoprotection agent with significant antioxidative activity, but the responsible antioxidative components are still unknown.
11)In our screening program of searching for antioxidative natural substances from traditional Chinese medicine, the ethanol extract of C. orchioides showed some antioxidative activities. Fractionation of this extract with a D-101 resin column chromatography confined the antioxidative activities to the 20% and 40% ethanol eluates. The concentrations exhibited 50% inhibition (IC 50 ) on hydroxyl radicals and superoxide anion radicals were 3.42 and 4.17 mg/ml for the 20% eluate, and 1.56 and 2.48 mg/ml for the 40% eluate, respectively. Further bioassay-guided analysis led to the isolation of 8 phenols and phenolic glycosides from these two active eluates of the C. orchioides extract. In this paper, we report the isolation and structural elucidation of a new orcinol glyco- , and superoxide anion radicals produced by xathine/xanthine oxidase systems, and compared with epigallocatechin gallate (EGCG), a known antioxidant.
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Results and DicussionThe ethanol extract of the plant was re-suspended in water and partitioned with petroleum ether (60-90°C). The water phase was filtered and the filtrate was loaded onto a D-101 resin column and then eluted sequentially with H 2 O followed by 20%, 40%, 60%, 80% and 95% aqueous EtOH. The fractions eluted from 20% and 40% EtOH were separately chromatographed on silica gel and Sephadex LH-20 to afford compounds 1-3 and compounds 4-8, respectively. Of the 8 compounds elucidated, the seven known compounds were subsequently identified as orcinol glucoside (1)4) curculigoside (4), curculigoside B (5), curculigoside C (6), 5) 2,6-dimethoxyl benzoic acid (7), 7) and syringic acid (8), 12,13) by comparison of spectral data with the data reported in the literature. The pu...