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Application of chromatography technology in the separation of active alkaloids from Hypecoum leptocarpum and their inhibitory effect on fatty acid synthase

Author:
Zhang, Qiulong, Luan, Guangxiang, Ma, Tao, Hu, Na, Suo, Yourui, Wang, Xiaoyan, Ma, Xiaofeng, Ding, Chenxu
Source:
Journal of separation science 2015 v.38 no.23 pp. 4063-4070
ISSN:
1615-9306
Subject:
Western blotting, alkaloids, breast neoplasms, carbon, chemical structure, countercurrent chromatography, cytotoxicity, electrospray ionization mass spectrometry, fatty-acid synthase, high performance liquid chromatography, infrared spectroscopy, neoplasm cells, nuclear magnetic resonance spectroscopy, reversed-phase liquid chromatography, spectral analysis, stable isotopes
Abstract:
A method that involved the combination of pH‐zone‐refining counter‐current chromatography and semipreparative reversed‐phase liquid chromatography has been established for the preparative separation of alkaloids from Hypecoum leptocarpum. From 1.2 g of crude sample, 31 mg N‐feruloyltyramine, 27 mg oxohydrastinine, 47 mg hydroprotopine, 25 mg leptopidine, and 18 mg hypecocarpine have been obtained. The structure of the new compound, hypecocarpine, is confirmed based on the analysis of spectroscopic data, including NMR, UV, and IR spectroscopy and positive electrospray ionization mass spectrometry. The known chemical structures were characterized on the basis of ¹H and ¹³C NMR spectroscopy. The purities of the five alkaloids are all over 92.7% as determined by high‐performance liquid chromatography. The alkaloids’ cytotoxicity in breast cancer cells is assessed by using a Cell Counting Kit assay and their inhibitory effect on fatty acid synthase expression is assessed by a Western blot assay. These results suggest that leptopidine could suppress growth and induce cytotoxicity in breast cancer cells and that the cytotoxicity of leptopidine may be related to its inhibitory effect on fatty acid synthase expression.
Agid:
4791220