title: Purification and characterization of aporphine alkaloids from leaves of Nelumbo nucifera Gaertn and their effects on glucose consumption in 3T3-L1 adipocytes.
abstract: Aporphine alkaloids from the leaves of Nelumbo nucifera Gaertn are substances of great interest because of their important pharmacological activities, particularly anti-diabetic, anti-obesity, anti-hyperlipidemic, anti-oxidant, and anti-HIV's activities. In order to produce large amounts of pure alkaloid for research purposes, a novel method using high-speed counter-current chromatography (HSCCC) was developed. Without any initial cleanup steps, four main aporphine alkaloids, including 2-hydroxy-1-methoxyaporphine, pronuciferine, nuciferine and roemerine were successfully purified from the crude extract by HSCCC in one step. The separation was performed with a simple two-phase solvent system composed of n-hexane-ethyl acetate-methanol-acetonitrile-water (5:3:3:2.5:5, v/v/v/v/v). In each operation, 100 mg crude extracts was separated and yielded 6.3 mg of 2-hydroxy-1-methoxyaporphine (95.1% purity), 1.1 mg of pronuciferine (96.8% purity), 8.5 mg of nuciferine (98.9% purity), and 2.7 mg of roemerine (97.4%) respectively. The chemical structure of four aporphine alkaloids are identified by means of electrospray ionization MS (ESI-MS) and nuclear magnetic resonance (NMR) analysis. Moreover, the effects of four separated aporphine alkaloids on insulin-stimulated glucose consumption were examined in 3T3-L1 adipocytes. The results showed that 2-hydroxy-1-methoxyaporphine and pronuciferine increased the glucose consumption significantly as rosiglitazone did.
authors.affiliation: School of Life Science, Yantai University, 30 Qinquan Road, Yantai 264005, China. chengjun-ma@163.com.
authors.forename: Chengjun
authors.initials: C
authors.lastname: Ma
authors.affiliation: School of Life Science, Yantai University, 30 Qinquan Road, Yantai 264005, China. wangjinjun19890522@163.com.
authors.forename: Jinjun
authors.initials: J
authors.lastname: Wang
authors.affiliation: School of Pharmacy, Yantai University, 30 Qinquan Road, Yantai 264005, China. chu_hong_mei@sina.com.
authors.forename: Hongmei
authors.initials: H
authors.lastname: Chu
authors.affiliation: School of Life Science, Yantai University, 30 Qinquan Road, Yantai 264005, China. mfsaih67@163.com.
authors.forename: Xiaoxiao
authors.initials: X
authors.lastname: Zhang
authors.affiliation: School of Life Science, Yantai University, 30 Qinquan Road, Yantai 264005, China. zhenhuawang@tom.com.
authors.forename: Zhenhua
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, China. hlwang@nwipb.cas.cn.
authors.forename: Honglun
authors.initials: H
authors.lastname: Wang
authors.affiliation: School of Life Science, Yantai University, 30 Qinquan Road, Yantai 264005, China. lgleegang@sohu.com.
authors.forename: Gang
authors.initials: G
authors.lastname: Li
mesh_terms: D041721:3T3-L1 Cells; D000085:Acetates / Q000737:chemistry; D000097:Acetonitriles / Q000737:chemistry; D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism; D000475:Alkenes / Q000737:chemistry; D000818:Animals; D001060:Aporphines / Q000302:isolation & purification* / Q000494:pharmacology*; D002851:Chromatography, High Pressure Liquid; D003377:Countercurrent Distribution; D005947:Glucose / Q000378:metabolism*; D007328:Insulin / Q000494:pharmacology; D009682:Magnetic Resonance Spectroscopy; D000432:Methanol / Q000737:chemistry; D051379:Mice; D031653:Nelumbo / Q000737:chemistry*; D018515:Plant Leaves / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D014867:Water / Q000737:chemistry
chemical_list: D000085:Acetates; D000097:Acetonitriles; D000475:Alkenes; D001060:Aporphines; D007328:Insulin; D012997:Solvents; D014867:Water; C081279:aporphine; C007650:ethyl acetate; C117224:1-hexene; D005947:Glucose; D000432:Methanol; C032159:acetonitrile