title: Ethyl acetate-n-butanol gradient solvent system for high-speed countercurrent chromatography to screen bioactive substances in okra.
abstract: High-speed countercurrent chromatographic separation (HSCCC) possesses the property of zero-loss of sample, which is very useful for the screening of bioactive components. In the present study, the ethyl acetate-n-butanol gradient HSCCC solvent system composed of n-hexane-ethyl acetate-n-butanol-water was investigated for the screening of bioactive substances. To screen the antiproliferative compounds in okra extract, we used the stationary phase ethyl acetate-n-butanol-water (1:1:10) as the stationary phase, and eluted the antiproliferative components by 6-steps of gradient using mobile phases n-hexane-ethyl acetate (1:2), n-hexane-ethyl acetate (1:4), n-hexane-ethyl acetate (0:4), n-butanol-ethyl acetate (1:4) n-butanol-ethyl acetate (1:2), n-butanol-ethyl acetate (2:2), and n-butanol-ethyl acetate (2:1). The fractions collected from HSCCC separation with the gradient solvent system were assayed for antiproliferative activity against cancer cells. Bioactive components were identified: a major anti-cancer compound, 4'-hydroxy phenethyl trans-ferulate, with middle activity, and a minor anti-cancer compound, carolignan, with strong activity. The result shows that the gradient solvent system is potential for the screening of bioactive compounds from natural products.
authors: Institute of Food Chemistry, Zhejiang Gongshang University, 149 Jiaogong Road, Hangzhou, Zhejiang 310012, China.
authors: Hao
authors: H
authors: Ying
authors: Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China.
authors: Heyuan
authors: H
authors: Jiang
authors: Institute of Food Chemistry, Zhejiang Gongshang University, 149 Jiaogong Road, Hangzhou, Zhejiang 310012, China.
authors: Huan
authors: H
authors: Liu
authors: Institute of Food Chemistry, Zhejiang Gongshang University, 149 Jiaogong Road, Hangzhou, Zhejiang 310012, China.
authors: Fangjuan
authors: F
authors: Chen
authors: Institute of Food Chemistry, Zhejiang Gongshang University, 149 Jiaogong Road, Hangzhou, Zhejiang 310012, China. Electronic address: qizhendu@163.com.
authors: Qizhen
authors: Q
authors: Du
mesh_terms: D028483:Abelmoschus / Q000737:chemistry*; D000970:Antineoplastic Agents / Q000032:analysis / Q000302:isolation & purification* / Q000494:pharmacology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003377:Countercurrent Distribution / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D010936:Plant Extracts / Q000032:analysis / Q000302:isolation & purification* / Q000494:pharmacology; D012997:Solvents / Q000737:chemistry
chemical_list: D000970:Antineoplastic Agents; D010936:Plant Extracts; D012997:Solvents
keywords: Anti-cancer; Bioactive compounds; Gradient HSCCC solvent system; Okra; Screening