高速逆流色谱法从内生真菌毕赤酵母 Ppf9 中制备分离海松酸。
Preparative separation of helvolic acid from the endophytic fungus Pichia guilliermondii Ppf9 by high-speed counter-current chromatography.
机构信息
Department of Plant Pathology, College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
出版信息
World J Microbiol Biotechnol. 2012 Mar;28(3):835-40. doi: 10.1007/s11274-011-0878-0. Epub 2011 Sep 9.
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 25°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, (1)H-NMR and (13)C-NMR spectra.
高速逆流色谱（HSCCC）首次应用于从与药用植物云南重楼相关的内生真菌毕赤酵母 Ppf9 的粗提物中制备分离羊毛甾酸。两相溶剂系统由正己烷-乙酸乙酯-甲醇-水（4.5:4.5:5.0:5.0，v/v）组成，并添加磷酸（0.2%，v/v）。分离柱的旋转速度、流动相的流速和仪器的分离温度分别为 800 rpm、3 ml min(-1)和 25°C。通过 HSCCC 在 4 小时的分离过程中，从 450 mg 的粗提物中成功获得了约 6.8 mg 的羊毛甾酸，根据 HPLC 分析，其纯度达到 93.2%。通过 MS、（1）H-NMR 和（13）C-NMR 谱对产物进行了进一步表征。
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