title: Preparative isolation and purification of three rotenoids and one isoflavone from the seeds of Millettia pachycarpa Benth by high-speed counter-current chromatography.
abstract: Both analytical and preparative high-speed counter-current chromatography (HSCCC) were used to isolate and separate chemical bioactive constituents from the seeds of Millettia pachycarpa Benth, a famous traditional Chinese medicine. Three rotenoids and one isoflavone were successfully purified for the first time by HSCCC with a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (HEMWat) (1:0.8:1:0.6, v/v/v/v). The separation parameters were first performed on the analytical HSCCC and optimized conditions were then scaled up to preparative HSCCC. The separation produced 160.2 mg tephrosin, 14.6 mg 4',5'-dimethoxy-6,6-dimethylpyranoisoflavone, 109.4 mg deguelin, 6.7 mg 6a,12a-dehydrodeguelin with respective purities of 95, 93, 95, 95%, in one single run from 400 mg crude extract of the seeds of M. pachycarpa Benth. The purity of the isolated compounds was analyzed by high-performance liquid chromatography (HPLC) and their structures were identified by electrospray ionization mass spectrometry (ESI-MS); (1)H nuclear magnetic resonance ((1)H NMR) and (13)C nuclear magnetic resonance ((13)C NMR) analysis. This paper is an excellent example of the role that CCC is playing in isolating active compounds for pre-clinical trials of new chemical entities, even when scaling up between centrifuges from different manufacturers.
authors: State Key Laboratory of Biotherapy, West China Hospital and College of Chemical Engineering, Sichuan University, Chengdu 610041, China.
authors: Haoyu
authors: H
authors: Ye
authors: Lijuan
authors: L
authors: Chen
authors: Yanfang
authors: Y
authors: Li
authors: Aihua
authors: A
authors: Peng
authors: Afu
authors: A
authors: Fu
authors: Hang
authors: H
authors: Song
authors: Minghai
authors: M
authors: Tang
authors: Houding
authors: H
authors: Luo
authors: Youfu
authors: Y
authors: Luo
authors: Yongbin
authors: Y
authors: Xu
authors: Jianyou
authors: J
authors: Shi
authors: Yuquan
authors: Y
authors: Wei
mesh_terms: D002851:Chromatography, High Pressure Liquid; D002867:Chromones / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D003377:Countercurrent Distribution; D007529:Isoflavones / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D009682:Magnetic Resonance Spectroscopy; D033641:Millettia / Q000737:chemistry*; D011714:Pyrans / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D012402:Rotenone / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D012639:Seeds / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D002867:Chromones; D007529:Isoflavones; D011714:Pyrans; D012402:Rotenone; C480848:tephrosin; C107676:deguelin