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Int J Mol Sci. 2014 Feb 26;15(3):3481-94. doi: 10.3390/ijms15033481.

Purification and characterization of aporphine alkaloids from leaves of Nelumbo nucifera Gaertn and their effects on glucose consumption in 3T3-L1 adipocytes.

Author information

  • 1School of Life Science, Yantai University, 30 Qinquan Road, Yantai 264005, China. chengjun-ma@163.com.
  • 2School of Life Science, Yantai University, 30 Qinquan Road, Yantai 264005, China. wangjinjun19890522@163.com.
  • 3School of Pharmacy, Yantai University, 30 Qinquan Road, Yantai 264005, China. chu_hong_mei@sina.com.
  • 4School of Life Science, Yantai University, 30 Qinquan Road, Yantai 264005, China. mfsaih67@163.com.
  • 5School of Life Science, Yantai University, 30 Qinquan Road, Yantai 264005, China. zhenhuawang@tom.com.
  • 6Key Laboratory of Tibetan Medicine Research, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810001, China. hlwang@nwipb.cas.cn.
  • 7School of Life Science, Yantai University, 30 Qinquan Road, Yantai 264005, China. lgleegang@sohu.com.

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.

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