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Isolation and purification of macrocyclic components from Penicillium fermentation broth by high‐speed counter‐current chromatography
Author(s) -
Gao Xiang,
Zhuang Rongqiang,
Guo Jiannan,
Bao Jian,
Fang Meijuan,
Liu Yan,
Xu Pengxiang,
Zhao Yufen
Publication year - 2010
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200900630
Subject(s) - countercurrent chromatography , chemistry , chromatography , ethyl acetate , extraction (chemistry) , high performance liquid chromatography , fermentation , partition coefficient , fraction (chemistry) , organic chemistry
In this paper, high‐speed counter‐current chromatography (HSCCC), assisted with ESI‐MS, was first successfully applied to the preparative separation of three macrolide antibiotics, brefeldin A (12.6 mg, 99.0%), 7′‐ O ‐formylbrefeldin A (6.5 mg, 95.0%) and 7′‐ O ‐acetylbrefeldin A (5.0 mg, 92.3%) from the crude extract of the microbe Penicillium SHZK‐15. Considering the chemical nature and partition coefficient ( K ) values of the three target compounds, a two‐step HSCCC isolation protocol was developed in order to obtain products with high purity. In the two‐step method, the crude ethyl acetate extract was first fractionated and resulted in two peak fractions by HSCCC using solvent system n ‐hexane/ethyl acetate/methanol/water (HEMWat) (3:7:5:5 v/v/v/v), then purified using solvent systems HEMWat (3:5:3:5 v/v/v/v) and HEMWat (7:3:5:5 v/v/v/v) for each fraction. The purities and structures of the isolated compounds were determined by HPLC, X‐ray crystallography, ESI‐MS and NMR. The results demonstrated that HSCCC is a fast and efficient technique for systematic isolation of bioactive compounds from the microbes.