title: Separation of flavonoids from Millettia griffithii with high-performance counter-current chromatography guided by anti-inflammatory activity.
abstract: Millettia griffithii is a unique Chinese plant located in the southern part of Yunnan Province. Up to now, there is no report about its phytochemical or related bioactivity research. In our previous study, the n-hexane crude extract of Millettia griffithii revealed significant anti-inflammatory activity at 100 μg/mL, inspiring us to explore the anti-inflammatory constituents. Four fractions (I, II, III, and A) were fractionated from n-hexane crude extract by high-performance counter-current chromatography with solvent system composed of n-hexane/ethyl acetate/methanol/water (8:9:8:9, v/v) and then were investigated for the potent anti-inflammatory activity. Fraction A, with the most potent inhibitory activity was further separated to give another four fractions (IV, V, VI, and B) with solvent system composed of n-hexane/ethyl acetate/methanol/water (8:4:8:4, v/v). Compound V and fraction B exhibited remarkable anti-inflammatory activity with nitric oxide inhibitory rate of 80 and 65%, which was worth further fractionation. Then, three fractions (VII, VIII, and IX) were separated from fraction B with a solvent system composed of n-hexane/ethyl acetate/methanol/water (8:1:8:1, v/v), with compound VIII demonstrating the most potent inhibitory activity (80%). Finally, the IC50 values of compound V and VIII were tested as 38.2 and 14.9 μM. The structures were identified by electrospray ionization mass spectrometry and(1)H and (13)C NMR spectroscopy.
authors.affiliation: State key laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu, China.
authors.forename: Huan
authors.initials: H
authors.lastname: Tang
authors.forename: Bo
authors.initials: B
authors.lastname: Wu
authors.forename: Kai
authors.initials: K
authors.lastname: Chen
authors.forename: Heying
authors.initials: H
authors.lastname: Pei
authors.forename: Wenshuang
authors.initials: W
authors.lastname: Wu
authors.forename: Liang
authors.initials: L
authors.lastname: Ma
authors.forename: Aihua
authors.initials: A
authors.lastname: Peng
authors.forename: Haoyu
authors.initials: H
authors.lastname: Ye
authors.forename: Lijuan
authors.initials: L
authors.lastname: Chen
mesh_terms: D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003377:Countercurrent Distribution; D004305:Dose-Response Relationship, Drug; D005419:Flavonoids / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors / Q000494:pharmacology; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D033641:Millettia / Q000737:chemistry*; D009569:Nitric Oxide / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D010936:Plant Extracts / Q000737:chemistry* / Q000494:pharmacology; D021241:Spectrometry, Mass, Electrospray Ionization; D013329:Structure-Activity Relationship
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D005419:Flavonoids; D008070:Lipopolysaccharides; D010936:Plant Extracts; D009569:Nitric Oxide
keywords: Anti-inflammatory activity; High-performance counter-current chromatography; Millettia griffithii; Nitric oxide production