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海产品贝毒素大田软海绵酸ELISA及HPLC-MS/MS检测方法的研究
  • ISSN号:1002-6630
  • 期刊名称:《食品科学》
  • 时间:0
  • 分类:TS254.7[轻工技术与工程—水产品加工及贮藏工程;轻工技术与工程—食品科学与工程]
  • 作者机构:[1]吉林大学人兽共患病研究所,人兽共患病教育部重点实验室,吉林长春130062, [2]吉林出入境检验检疫局,吉林长春130062, [3]福清出入境检验检疫局,福建福清350300
  • 相关基金:基金项目:国家自然科学基金项目(30671762);国家质检总局项目(2003IK044;2008IK003)
中文摘要:

为了检测海产品贝毒大田软海绵酸,保障其食用安全,利用活泼酯法将小分子OA与载体蛋白偶联,免疫BALB/c小鼠,细胞融合技术建立分泌抗OA的杂交瘤细胞株,并对其各种特性进行分析;小鼠腹水法大量生产抗体,纯化后建立ELISA方法,同时建立OA的高效液相色谱-串联质谱(HPLC-MS)检测法,并对部分市售海产品进行了实际检测,ELISA方法标准曲线为y=-34.212x+83.49,相关系数为0.9784,线性范围0.4~25μg/L,灵敏度0.18μg/L;HPLC-MS/MS检测方法标准曲线y=193.07x-780.6(Q1/Q3:m/z827.4~m/z723.5)和y=83.021x-335.6(Q1/Q3:m/z827.4~m/z809.5),R2均为0.9991,线性范围10~800μg/L,灵敏度小于2μg/L,平均RSD为4.34%。在检测的实际样品中两种样品ELISA呈阳性反应,其中一种经过了HPLC-MS/MS的验证。所建立的ELISA及HPLC-MS检测方法均可用于海产品腹泻性贝毒OA限量标准检测,为进出口海产品OA标准方法的建立提供实验基础。

英文摘要:

The aim of this experiment was to establish rapid detection methods of okadaic acid (OA) in shellfish in order to ensure edible safety. The OA was coupled with human IgG as immunity antigen and with BSA as detection antigen by active ester method. The Balb/c mice were inoculated with the prepared antigen. The hybridoma secreting monoclonal antibody against OA was constructed by the cell-fusion technology and the specialities of the monoclonai antibody were analyzed. The ELISA method was established for detecting OA after ascites was prepared and purified. At the same time, the HPLC-MS/MS method was also set up. Some shellfish samples were detected by the two methods. The linear regression equation of ELISA method is y =- 34.212x + 83.49 (y is optical density at 490-nm wavelength, and x is OA concentration,μg/L) with the determination coefficient of 0.9784, which shows good linearity over the concentration range of 0.4 to 25 μg/L, and the detection limit to OA is 0.18μg/L. The linear regression equations of HPLC-MS/MS method are y = 193.07x - 780.6 (Q1/Q3: m/z 827.4-m/z 723.5) and y = 83.021x - 335.6 (Q1/Q3: m/z 827.4-m/z 809.5), where y is peak height, and x is OA concentration, μg/L. Both the equations have the same determination coefficients of 0.9991, show good linearity over the concentration range of 10 to 800 μg/L. The detection limit to OA is less than 2 μg/L and the stand average RSD is 4.34%. Two shellfish samples were the positive result by ELISA method, and one of them was verifed by HPLC-MS/MS. The two methods both can be used for detecting OA and provide experimental foundation to establish standard detection method for OA in shellfish.

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期刊信息
  • 《食品科学》
  • 北大核心期刊(2011版)
  • 主管单位:中国商业联合会
  • 主办单位:北京市食品研究所
  • 主编:孙勇
  • 地址:北京西城区禄长街头条4号
  • 邮编:100050
  • 邮箱:foodsci@126.com
  • 电话:010-83155446-8006
  • 国际标准刊号:ISSN:1002-6630
  • 国内统一刊号:ISSN:11-2206/TS
  • 邮发代号:2-439
  • 获奖情况:
  • 国家“双效”期刊,1986年原商业部重大成果三等奖,1997年国内贸易部优秀科技期刊三等奖,第三届中国出版政府奖提名奖,第三届中国出版政府奖提名奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),美国工程索引,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),英国英国皇家化学学会文摘,英国食品科技文摘,中国北大核心期刊(2000版)
  • 被引量:115579