网状原角藻中虾夷扇贝毒素提取与纯化  

Extraction and purification of yessotoxin in the dinoflagellate Protoceratium reticulatum

作　　者：姚敬元 金薇 陈文博 宫长宝 贾宝林 隋世峰 冯少姝 韩蕾 梁玉波[1,3] 吕颂辉 YAO Jing-yuan;JIN Wei;CHEN Wen-bo;GONG Chang-bao;JIA Bao-lin;SUI Shi-feng;FENG Shao-shu;HAN Lei;LIANG Yu-bo;LV Song-hui(National Marine Environmental Monitoring Center,Dalian 116023,China;Jinan University,Guangzhou 510632,China;Dalian key laboratory of algal toxin,Dalian 116023,China;Dalian modern agricultural production development service center,Dalian 116023,China;Dalian University of Technology,Dalian 116024,China)

机构地区：[1]国家海洋环境监测中心,辽宁大连116023 [2]暨南大学,广东广州510632 [3]大连市藻毒素重点实验室,辽宁大连116023 [4]大连市现代农业生产发展服务中心,辽宁大连116023 [5]大连理工大学,辽宁大连116024

出　　处：《海洋环境科学》2021年第1期101-106,共6页Marine Environmental Science

基　　金：国家重点研发计划项目(2016YFF0201104,2017FYC1404303);国家自然科学基金项目(41576120,41276099);国家海洋局海洋赤潮灾害立体监测技术与应用重点实验室开放基金项目(MAYTHAB201605);国家海洋局近岸海域生态环境重点实验室开发基金项目(201613)。

摘　　要：通过批量培养可产生虾夷扇贝毒素的网状原角藻,对获得的藻泥细胞超声破碎后,用甲醇提取;用大孔吸附树脂(HP20)吸附网状原角藻分泌于培养液中的胞外毒素,用70%的乙醇洗脱吸附于树脂上的毒素。采用高效液相色谱-串联质谱法,在网状原角藻毒素提取液中,发现9种虾夷扇贝毒素及其类似物,其中虾夷扇贝毒素YTX占总量95.1%,41YTX占3.65%,其他类似物共占1.25%。采用高效制备液相色谱分离纯化5次,获得的虾夷扇贝毒素经高分辨质谱法确认其分子量为1142.4747,分子式为C_(55)H_(82)O_(21)S_(2),定量核磁法检测其纯度超过99%,满足虾夷扇贝毒素标准物质制备要求。Marine dinoflagellate Protoceratium reticulatum producing yessotoxins was mass cultured.Then obtained algal cells were extracted by ultrasonic crushing with methanol for yessotoxins.Macroporous adsorption resin(HP20)was used to adsorb the extracellular toxin secreted in the culture medium,and the yessotoxins adsorbed on the resin were eluted with 70%ethanol.Nine of yessotoxin and their analogues were detected by high performance liquid chromatography-tandem mass spectrometry in extracts.The toxin of yessotoxin accounted for 95.1%of the total toxin group in the extracts,derivative 41YTX accounted for 3.65%,and other trace analogues accounted for 1.25%.Yessotoxin was purified through high performance preparation liquid chromatography for 5 times.The molecular weight and molecular formula was confirmed as 1142.4747 and C_(55)H_(82)O_(21)S_(2),respectively,by high resolution mass spectrometry.The purity of yessotoxin detected by quantitative nuclear magnetic method was more than 99%,meeting the requirement for preparation of yessotoxin standard material.

关 键 词：网状原角藻 毒素提取与纯化 高纯度虾夷扇贝毒素 

分 类 号：X174[环境科学与工程—环境科学]