高速逆流色谱法快速分离花脸香蘑中细胞毒活性化合物

Rapid is olation of cytotoxic compound from Lepista sordida by high-speed countercurent chromatography

首次应用高速逆流色谱技术(High-speed countercurrent chromatography,HSCCC),选择石油醚-乙酸乙酯-甲醇-水(9:1:9:1,v/v)作为溶剂系统对花脸香蘑固体发酵乙酸乙酯粗提物进行了分离纯化,在2 mL/min流速、900 r/min转速和25℃分离条件下,固定相保留率达到76.7%,从100 mg粗提物中一次性分离得到活性化合物4.3 mg,纯度为95.6%,回收率为97%,并通过MS和NMR等波谱技术鉴定所分离得到的活性化合物为1-羟基-6-甲基-9,10-蒽醌。细胞毒活性结果表明,1-羟基-6-甲基-9,10-蒽醌对人肺癌A549、人骨肉瘤MG63、人结肠癌Sw480和人肝癌Hep G2细胞具有中等细胞毒活性,IC_(50)分别为48.2、114.8、108.1、112.3μg/mL。1-羟基-6-甲基-9,10-蒽醌为首次从花脸香蘑中分离获得。

The crude sample of solid cultures of the fungus Lepista sordida was purified by high speed countercurrent chromatography with petroleum ether/ethyl acetate/methanol/water(9:1:9:1, v/v) as the optimized two-phase solvent system. Under the flow rate 2.0 mL/min, the roltation speed 900 r/min,separation temperature 25 ℃, the retention rate of the stationary phase reached 76.7%. 4.3 mg aliquot of 1-hydroxy-6-methyl-9,10-anthraquinone was separated from 100 mg of the crude sample. The purity of 1-hydroxy-6-methyl-9,10-anthraquinone was 95.6% with the recovery of 97%, as determined by high performance liquid chromatrography. The chemical structure was identified primarily by NMR and MS experiments. 1-hydroxy-6-methyl-9,10-anthraquinone showed modest cytotoxic activities against four human tumor cells. 1-hydroxy-6-methyl-9,10-anthraquinone was isolated from the solid cultures of the fungus Lepista sordida for the first time.