Isolation and purification of macrocyclic components from Penicillium fermentation broth by high‐speed counter‐current chromatography |
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Authors: | Xiang Gao Rongqiang Zhuang Jiannan Guo Jian Bao Meijuan Fang Yan Liu Pengxiang Xu Yufen Zhao |
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Affiliation: | 1. Department of Chemistry and The Key Laboratory for Chemical Biology of Fujian Province, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, P. R. China;2. Department of Medicine, Medical College, Xiamen University, Xiamen, P. R. China;3. The Key Laboratory for Bioorganic Phosphorus Chemistry and Chemical Biology (Ministry of Education), Department of Chemistry, School of Sciences, Tsinghua University, Beijing, P. R. China |
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Abstract: | 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. |
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Keywords: | Brefeldin A ESI‐MS High‐speed counter‐current chromatography Macrolide antibiotics Preparative chromatography |
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