期刊
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
卷 28, 期 3, 页码 835-840出版社
SPRINGER
DOI: 10.1007/s11274-011-0878-0
关键词
Preparative separation; Helvolic acid; Endophytic fungus; Pichia guilliermondii Ppf9; High-speed counter-current chromatography
资金
- Hi-Tech R&D Program of China [2011AA10A202]
- National Basic Research Program of China [2010CB126105]
- National Natural Science Foundation of China [30871662, 31071710]
High-speed counter-current chromatography (HSCCC) was applied for preparative separation of helvolic acid from the crude extract of the endophytic fungus Pichia guilliermondii Ppf9, associated with the medicinal plant Paris polyphylla var. yunnanensis for the first time. The two-phase solvent system consisted of n-hexane-ethyl acetate-methanol-water (4.5:4.5:5.0:5.0, v/v) appending with phosphoric acid (0.2%, v/v) was employed. The revolution speed of the separation column, flow rate of the mobile phase and separation temperature of the apparatus were 800 rpm, 3 ml min(-1) and 25A degrees C, respectively. About 6.8 mg of helvolic acid was successfully obtained from 450 mg of the crude extract by HSCCC within 4 h separation procedure, and its purity reached to 93.2% according to the HPLC analysis. The product was further characterized by MS, H-1-NMR and C-13-NMR spectra.
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