一种脱氧雪腐镰刀菌烯醇制备工艺的研究

Research on a Preparation Process of Deoxynivalenol

脱氧雪腐镰刀菌烯醇(DON)是镰刀菌属产生的一种真菌毒素,具有细胞毒性,可导致胚胎发育畸形,严重威胁人畜健康。为了获得大量的高纯度DON用于毒理学及脱毒研究,本研究开发了一种操作简单、经济高效的DON分离纯化方法:抽提:采用84%的乙腈水溶液从禾谷镰刀菌PH-1的大米培养物中提取DON获得DON粗毒素;净化:使用石油醚和饱和食盐水净化DON粗毒素;精制:使用乙酸乙酯:石油醚(1∶1,V/V)洗涤净化后的粗毒素,再利用乙酸乙酯和饱和食盐水之间的极性差异,通过液液分配的方式萃取DON,进一步去除杂质;纯化:使用制备型高效液相色谱纯化DON,获得DON纯品。该方法可以从每千克PH-1大米培养物中获得61.4 mg DON纯品,其优势是经过精制的DON纯度已经达到80%以上,可以满足常规研究的需求,如需通过制备液相色谱进一步纯化,可显著降低色谱柱损耗,降低经济成本。

Deoxynivalenol(DON)is a mycotoxin produced by Fusarium.It is cytotoxic and can cause embryo deformity,which seriously threatens human and animal health.To provide enough high-purity DON for scientific research in toxicology and detoxification,we developed a simple,economical and effective process for the separation and purification of DON.1.Extraction:84%acetonitrile aqueous solution was used to extract DON from the rice culture of Fusarium graminearum PH-1 to obtain DON crude extract.2.Cleaning:Petroleum ether and saturated saline aqueous solution were used to remove the impurity from the DON crude extract.3.Refining:The product obtained in step 2 was washed by ethyl acetate:petroleum ether(1∶1 of V/V).Then more impurity was removed by liquid-liquid distribution based on the polarity difference between ethyl acetate and saturated saline aqueous solution.4.Purification:Helped by preparative high performance liquid chromatography DON in high purity was obtained.With this method 61.36 mg highly purified DON was obtained from one kilogram PH-1 culture.Its main feature is that the purity of the refined DON has reached 80%,which could be directly applied in conventional scientific research.If further purification needed via preparative liquid chromatography,the column loss can be remarkably reduced,therefore the economic cost decreases.