文章摘要
林华娟,秦小明,章超桦,长岛裕二.扇贝中麻痹性毒素的提取与分离纯化[J].水产学报,2010,34(3):428~434
扇贝中麻痹性毒素的提取与分离纯化
Extraction and purification of paralytic shellfish poison from scallop
投稿时间:2009-10-04  修订日期:2009-12-09
DOI:10.3724/SP.J.1231.2010.06637
中文关键词: 扇贝  麻痹性毒素(PSP毒素)  分离纯化  膝沟藻毒素(GTXs)
英文关键词: scallop  paralytic shellfish poison (PSP)  purification  gonyatoxins (GTXs)
基金项目:农业部948项目(2006-G42);现代农业产业技术体系建设专项
作者单位E-mail
林华娟 广东海洋大学 huajuanlin@163.com 
秦小明 广东海洋大学  
章超桦 广东海洋大学  
长岛裕二 日本东京海洋大学  
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中文摘要:
      为了获得麻痹性毒素标准品,以毒化扇贝为试验材料,对扇贝中的麻痹性毒素(PSP毒素)进行了提取和分离纯化。以酸性80%乙醇溶液反复提取麻痹性毒素,得到了总毒性为6170 MU的毒素粗提液。毒素粗提液经过超滤、Bio-Gel P-2凝胶柱层析和Bio-Rex 70离子交换柱层析等二次层析柱分离纯化后得到以膝沟藻毒素群(GTXs)为主的纯化PSP毒素。HPLC分析结果显示,扇贝中主要含有膝沟藻毒素GTX3,GTX1、GTX2、GTX3、和GTX4的组成比例约为2∶4∶14∶1(以HPLC上的峰面积比例计)。纯化后的毒素可以作为标准物质用于GTX1-4的HPLC分析。
英文摘要:
      HPLC analysis for paralytic shellfish poison (PSP) in shellfish foods has now become a current method in the world, in which PSP standard is necessary. Unfortunately, commercial PSP standard has now become difficult for handling in china, as some purification technique of PSP has not been still breached, and the PSP substance has now been recognized into the list of chemical weapons. Preparing independently some PSP standard is necessary and will be a good way for solving the standard substance problem in china.  In this paper, extraction and purification of PSP was discussed, using a poisoned scallop sample as a material. The total toxicity of the crude PSP extract solution was 6 170 MU, while using acid 80% ethanol as an extractant. From the crude extract, PSP was purified via ultrafiltration, Bio-Gel P-2 column chromatography and Bio Rex 70 column chromatography, and a purified toxin with amount of 2 100 MU was obtained. The result of chromatography showed the toxin mainly contained gonyautoxin group (GTX1-4). HPLC analysis showed that all of the peaks is very sharp and clear, in which the ratio of GTX1, GTX2, GTX3 and GTX4 was 2∶4∶14∶1 (calculated as the ratio of peak area on HPLC), indicating that GTX3 is the major PSP composition in the poisoned scallop sample, and the purified toxin is available using as a standard for HPLC analysis. From the results of experiments, the technique for preparing PSP standard is effective and successful. In the other hand, the stability of the purified toxin was discussed, through comparing the HPLC analysis between the native toxin and that of the one treated with 0.1 mol/L HCl.
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