摘要
目的开发一种高纯度烟曲霉素的分离纯化工艺,并对烟曲霉素中的主要杂质进行分离纯化、结构鉴定。方法发酵液采用全料液萃取,以烟曲霉素收率为指标筛选最佳萃取溶剂,优化萃取条件,以甲醇和二氯甲烷混合溶液进行结晶,通过HPLC对烟曲霉素晶体中的主要杂质进行分离纯化,并采用核磁共振和质谱分析鉴定杂质结构。结果以甲基叔丁基醚为萃取溶剂得到的烟曲霉素收率最高,当甲基叔丁基醚:发酵液体积比为2:1时,萃取收率大于85%,再经过两次结晶可以得到纯度95%以上的烟曲霉素晶体,分离并鉴定了烟曲霉素晶体中的两个主要杂质。结论通过对烟曲霉素提取工艺研究,确定了较优的工艺条件,该方法具有易操作、纯度高、收率高、成本低的特点,适合工业化生产,烟曲霉素中两个主要杂质是烟曲霉素类似物,通过杂质鉴定为烟曲霉素质量控制和安全性评价提供支持。
Objective To develop a separation and purification process of high purity fumagillin, and to isolate and purify the main impurities in fumagillin and identify their structures. Methods In order to screen and optimize the extraction solvent, the fermentation broth was extracted with whole broth solvent extraction(WBE), and the yield of fumagillin was used as the index. The crystallization was carried out in a mixture of methanol and dichloromethane. The main impurities in fumagillin crystals were separated and purified by HPLC, and the structures of impurities were identified by NMR and mass spectrometry. Results The extraction solvent was methyl tert-butyl ether(MTBE), and the extraction yield was higher than 85% when the volume ratio of MTBE:fermentation broth was 2:1. After two crystallization, the crystals with purity above 95% could be obtained. The two main impurities in the fumagillin crystals were separated and identified. Conclusion Through the study on the extraction technology of fumagillin, the optimum technological conditions are determined. The method has the characteristics of easy operation, high purity, high yield and low cost, which is suitable for industrial production. The two main impurities are the analogues of fumagillin. The impurity identification provides support for quality control and safety evaluation of fumagillin.
作者
李建宋
李亚
王继栋
郝之奎
薛长艳
LI Jian-song;LI Ya;WANG Ji-dong;HAO Zhi-kui;XUE Chang-yan(Taizhou Vocational&Technical College,Zhejiang 318000,China;Zhejiang Hisun Pharmaceutical Co.,Ltd,Taizhou 318000,China;School of Life Sciences,Huzhou University,Zhejiang 313000,China)
出处
《中国医药生物技术》
2020年第4期398-403,共6页
Chinese Medicinal Biotechnology
关键词
烟曲霉素
分离纯化
杂质
结构鉴定
Fumagillin
Separation and purification
Impurities
Structure identification
