首次运用高速逆流色谱(HSCCC)技术从经表观遗传试剂诱导的曲霉属真菌Aspergullus versicolor的次级代谢产物中快速分离纯化得到二苯醚类化合物diorcinol,建立了快速分离制备杂色曲霉次级代谢产物中的二苯醚类化合物的方法.本研究首先对经过表观遗传试剂诱导的菌株D J013的发酵液用乙酸乙酯浸提,萃取富集二苯醚类成分,然后以石油醚-乙酸乙酯-甲醇-水(4:5:4:5,v/v)为两相溶剂系统进行高速逆流色谱分离纯化,上相为固定相,下相为流动相,流速5.0 mL/min,实验温度25℃,转速为800 rpm,检测波长为220 nm.对所得到的目标化合物经超高效液相色谱(UPLC)纯度分析,其纯度在97％以上.通过质谱、核磁等波谱技术鉴定所分离得到的目标化合物为二苯醚类化合物diorcinol.与前期研究中采用的柱色谱法、HPLC等多种方法相结合的长达48 h的制备周期相比,高速逆流色谱法仅需55 min,效率大大提高.该结果表明,本研究建立的高速逆流色谱方法可高效高纯度获得具有抗菌活性的二苯醚类化合物,将为二苯醚化合物的进一步研究提供高效制备方法.%Preparative high speed counter-current chromatography (HSCCC) was applied for the isolation and purification of a diorcinol from the fungus Aspergillus versicolor DJ013 regulated by epigenetic manipulation for the first time using stepwise elution with the two-phase solvent system composed of petroleum ether-ether acetate-methanol-water at (4:5:4:5,v/v).The preparative HSCCC separation was performed on crude sample yielding diorcinol in a one-step separation,with purities over 97％ as determined by UPLC.The temperature was 25 ℃ and the revolution speed was 800 rpm with a flow rate of 5.0mL/min.The eluent was continuously monitored by a UV monitor at 220 nm.The structure of this compound was identified by ESI-MS and NMR.Compared with the 48-hour preparation period combined with column chromatography and HPLC,the high speed countercurrent chromatography only needs 55 min,and the efficiency is greatly improved.Our results demonstrated that HSCCC was an efficient technique to separate diorcinol,which provide an approach to solve the problem of their sample availability for drug development.
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