[关键词]
[摘要]
本文主要研究分子层析柱对河鲀鱼肝脏中河豚毒素分离纯化效果,为河豚毒素分离纯化工艺提供参考。本实验以假睛东方鲀肝脏为原料,对肝脏中的TTX进行提取,然后利用柱层析技术,确定了中性氧化铝柱、活性炭柱、CM HF和D152弱酸性阳离子交换树脂柱的上样液最适pH、吸附率、回收率。并根据四种柱子的吸附率和净化回收率的结果综合评定,其中D152弱酸性阳离子交换柱吸附率和回收率分别为91.97%和93.98%,且该柱子纯化河豚毒素的效果好,损失率低,最后得到产物多,因此选择D152柱作为分离纯化河豚毒素粗提液纯化柱。最后以假睛东方鲀肝脏为原料,将肝脏匀浆得到的粗提液经D152柱纯化分离后,结晶析出得到河豚毒素粗品,纯度为80%。此研究为河豚毒素分离纯化工艺提供有益参考。
[Key word]
[Abstract]
The separation and purification effect of chromatographic column on tetrodotoxin was studied, which provided a reference for the separation and purification process of tetrodotoxin. The raw material for this experiment was the liver of Takifugu pseudommus, TTX was extracted from the liver, and the column chromatography technology by the loading of neutral alumina column, activated carbon column, CM HF and D152 weak acid cation exchange resin column was used to determine theoptimal pH, adsorption rate and recovery rate of liquid. According to the results of the adsorption rate and purification recovery rate of the four columns, the adsorption rate and recovery rate of the D152 weakly acidic cation exchange column were 91.97% and 93.98%, respectively, and the column was effective in purifying tetrodotoxin with low loss rate. Finally, by D152 column the crude tetrodotoxin extract was separated and purified. The raw material was Takifugu pseudommus liver. The crude extract obtained from liver homogenization was purified and separated by D152 column, and then was crystallized to obtain crude tetrodotoxin with a purity of 80%. This study provides a useful reference for the separation and purification process of tetrodotoxin.
[中图分类号]
[基金项目]
国家重点研发计划项目(2017YFC1600701)