Abstract
Objective: Prepare immunoaffinity column(IAC) for pre-processing Neurotoxic Shellfish Poisoning(NSP) samples, and to establish a method of IAC purification LC-MS/MS spectrometry for the NSP. Method:The IAC for the NSP was prepared by purification of PbTX-2 monoclonal antibody and agarose gel (Sepharose 4B) as carrier. The samples were extracted with 80% methanol aqueous solution, diluted with PBS buffer, purified and enriched by the IAC, and analyzed by LC-MS/MS. Meanwhile, the parameters of IAC such as sample solution and eluent were optimized. The results showed: The IAC for the NSP had a good purification effect on shellfish samples, reduced the interference of fat, protein, pigment, mucus and other impurities in shellfish matrix, and improved the recovery. In the range of 40~800 ng, the average recovery of standard addition was 82.95%~112.95%, and the relative standard deviation was 1.27%~2.94%. Conclusion: The IAC for the NSP can meet the needs of daily monitoring, with small matrix interference, strong purification effect and high sensitivity. It’s suitable for the analysis and determination of the NSP.
Publication Date
2-28-2020
First Page
84
Last Page
88
DOI
10.13652/j.issn.1003-5788.2020.02.015
Recommended Citation
Xin, DONG; Jing-miao, BAI; and Hai-yan, WU
(2020)
"The preparation and application of immunoaffinity column for neurotoxic shellfish poisoning,"
Food and Machinery: Vol. 36:
Iss.
2, Article 15.
DOI: 10.13652/j.issn.1003-5788.2020.02.015
Available at:
https://www.ifoodmm.cn/journal/vol36/iss2/15
References
[1] 李大志.以现代分析技术为基础的贝类毒素地理分布、季节变化及致毒机理研究[D].大连:中国科学院大连化学物理研究所,2005:2-10.
[2] 曹际娟,赵昕,郭皓,等.神经性贝类毒素[J].检验检疫科学,1999,6(9):56-58.
[3] 江涛,江天久.贝类体内麻痹性贝类毒素的提取方法研究[J].分析化学,2008,36(11):1 460-1 464.
[4] 李兆新,张婷婷,郭萌萌,等.贝类毒素固相吸附示踪检测技术研究进展[J].渔业科学进展,2016,37(10):162-165.
[5] 李芳,李雪梅,李献刚,等.贝类毒素检测方法研究概况[J].食品研究与开发,2015,36(23):185-186.
[6] 黄宗锈,林健,陈冠敏,等.麻痹性贝类毒素小鼠生物法检测中位数法与均数法差异性比较[J].中国卫生检验杂志,2018,18(2):369-370.
[7] 黎小正,黄玉柳,吴详庆,等.腹泻性贝类毒素生物法检测中的误差原因分析[J].微生物学杂志,2009,29(4):107-109.
[8] 刘淑娟,赵晓祥,程金平,等.快速检测软骨藻酸的间接ELisa方法[J].环境科学学报,2014,34(2):404-408.
[9] 陈剑刚,朱炳辉,梁素丹,等.固相萃取—高效液相色谱—串联质谱法测定贝类产品中麻痹性贝类毒素[J].中国卫生检验杂志,2016,26(1):4-8.
[10] 郭萌萌,彭吉星,吴海燕,等.麻痹性贝类毒素GTX1&4和GTX2&3标准样品的制备技术[J].中国渔业质量与标准,2019,9(2):62-69.
[11] 严忠雍,张小军,李奇富,等.免疫亲合净化—液相色谱—串联质谱法测定贝类中腹泻性贝类毒素[J].食品科学,2017,38(18):304-309.
[12] 岳亚军,朱波,伦丽丽,等.石房蛤毒素免疫亲和柱制备及柱效测试研究[J].食品安全质量检测学报,2019,10(14):4 566-4 571.
[13] 母清林,方杰,万汉兴,等.液相色谱—串联质谱法检测贝类产品中腹泻贝类毒素[J].分析化学研究简报,2011,39(1):111-114.
[14] 姚佳,王昕,张建新,等.免疫亲和柱—高效液相色谱法测定牛奶中氯霉素和玉米赤霉醇及其类似物[J].食品科学,2014,35(18):124-127.
[15] 岳亚军,张律,游杰,等.免疫亲和柱净化—高效液相色谱-串联质谱法测定鱼肉和肝脏中河鲀毒素[J].中国食品卫生杂志,2016,28(2):214-218.
[16] 于广磊,刘磊,于姬,等.免疫亲和色谱分离纯化PSP贝毒GTX2,3 的研究[J].中国卫生检验杂志,2011,21(10):2 424-2 426.
[17] 王里奇,于师宇,任洪林,等.抗神经性贝毒素PbTx-2单克隆抗体的制备及鉴定[J].食品科技,2011,36(4):299-303.
[18] 王莹,路晟,柳家鹏,等.黄曲霉M1免疫亲和柱的制备及使用条件优化[J].中国奶牛,2011(1):44-46.