title: Isolation and purification of series bioactive components from Hypericum perforatum L. by counter-current chromatography.
abstract: Counter-current chromatography (CCC) combined with pre-separation by ultrasonic solvent extraction was successively used for the separation of series bioactive compounds from the crude extract of Hypericum perforatum L. The petroleum ether extract was separated by the solvent system of n-heptane-methanol-acetonitrile (1.5:0.5:0.5, v/v) and n-heptane-methanol (1.5:1, v/v) in gradient elution, yielding a phloroglucinol compound, hyperforin with HPLC purity over 98%. The ethyl acetate extract was separated by using the solvent system composed of hexane-ethyl acetate-methanol-water (1:1:1:1 and 1:3:1:3, v/v) in gradient through both reverse phase and normal phase elution mode, yielding a naphthodianthrone compound, hypericin with HPLC purity about 95%. The n-butanol extract was separated with the solvent system composed of n-butanol-ethyl acetate-water (1:4:5 and 1.5:3.5:5, v/v) in elution and back-extrusion mode, yielding two of flavones, rutin and hyperoside, with HPLC purity over 95%. HPLC-MS, reference sample and UV spectrum were selectively used in separation to search for target compounds from HPLC-DAD profiles of different sub-extracts. The structures of isolated compounds were further identified by ESI-MS, ¹HNMR and ¹³CNMR.
authors.affiliation: Beijing Key Lab of Plant Resource Research and Development, Beijing Technology and Business University, Beijing 100037, China. caoxl@th.btbu.edu.cn
authors.forename: Xueli
authors.initials: X
authors.lastname: Cao
authors.forename: Qiaoe
authors.initials: Q
authors.lastname: Wang
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Ge
authors.initials: G
authors.lastname: Bai
authors.forename: Hong
authors.initials: H
authors.lastname: Ren
authors.forename: Chunming
authors.initials: C
authors.lastname: Xu
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Ito
mesh_terms: D000873:Anthracenes; D001643:Bridged Bicyclo Compounds / Q000737:chemistry / Q000302:isolation & purification; D005591:Chemical Fractionation; D002851:Chromatography, High Pressure Liquid; D003377:Countercurrent Distribution / Q000379:methods*; D044948:Flavonols / Q000737:chemistry / Q000302:isolation & purification*; D020902:Hypericum / Q000737:chemistry*; D013058:Mass Spectrometry; D019906:Nuclear Magnetic Resonance, Biomolecular; D010569:Perylene / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification; D010696:Phloroglucinol / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification; D010936:Plant Extracts / Q000737:chemistry*; D013729:Terpenes / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D000873:Anthracenes; D001643:Bridged Bicyclo Compounds; D044948:Flavonols; D010936:Plant Extracts; D013729:Terpenes; D010569:Perylene; C004965:hypericin; D010696:Phloroglucinol; C001654:hyperforin