固相萃取/超高效液相色谱-串联质谱法测定贝类中3种短裸甲藻毒素
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摘要
建立了同时测定贝类中3种短裸甲藻毒素的超高效液相色谱-串联质谱法。样品采用丙酮提取、C18固相萃取柱净化;以5 mmol/L乙酸铵(含0. 1%甲酸)-乙腈为流动相,ACQUITYTMUPLC BEH C18色谱柱进行分离,在多反应监测模式下进行测定,外标法定量。结果表明,3种目标物在1. 0～100μg/L质量浓度范围内线性良好,相关系数(r~2)不小于0. 996,方法定量下限为3. 0μg/kg,回收率为80. 0%～87. 5%,相对标准偏差为1. 3%～5. 4%。该方法前处理简单,灵敏度高,稳定性好,适用于贝类中3种短裸甲藻毒素的同时测定。
An ultra performance liquid chromatography-tandem mass spectrometric( UPLC-MS/MS) method was developed for the determination of 3 kinds of brevetoxins( BTX) in shellfish. The samples were extracted with acetone,and purified on a C18 solid phase extraction column. The separation of the analytes was performed on an ACQUITYTMUPLC BEH C18 column with 5 mmol/L ammoumium acetate containing 0. 1% formic acid-acetonitrile as mobile phase,and detected in multiple reaction monitoring( MRM) mode. The external standard method was used for quantitation. Results showed that there were good linear relationships for BTXs in the range of 1. 0-100 μg/L with correlation coefficients( r~2) not less than 0. 996. The quantitation limits of the method were 3. 0 μg/kg. The recoveries for the brevetoxins ranged from 80. 0% to 87. 5% with relative standard deviations of 1. 3%-5. 4%. The method was suitable for the determination of brevetoxins in shellfish with advantages of simple processing,high sensitivity and good stability.
An ultra performance liquid chromatography-tandem mass spectrometric( UPLC-MS/MS) method was developed for the determination of 3 kinds of brevetoxins( BTX) in shellfish. The samples were extracted with acetone,and purified on a C18 solid phase extraction column. The separation of the analytes was performed on an ACQUITYTMUPLC BEH C18 column with 5 mmol/L ammoumium acetate containing 0. 1% formic acid-acetonitrile as mobile phase,and detected in multiple reaction monitoring( MRM) mode. The external standard method was used for quantitation. Results showed that there were good linear relationships for BTXs in the range of 1. 0-100 μg/L with correlation coefficients( r~2) not less than 0. 996. The quantitation limits of the method were 3. 0 μg/kg. The recoveries for the brevetoxins ranged from 80. 0% to 87. 5% with relative standard deviations of 1. 3%-5. 4%. The method was suitable for the determination of brevetoxins in shellfish with advantages of simple processing,high sensitivity and good stability.
引文
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[7] GB/T 5009. 261-2016. Determination of Neurotoxic Shellfish Poisons in Shellfish. National Standards of the People's Re-public of China(贝类中神经性贝类毒素的测定.中华人民共和国国家标准).
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[10] Wang L Q. Preparation of Monoclonal Antibody Against Brevetoxin and Establishment of ELISA Detected Method. Chang-chun:Jilin University(王里奇.短裸甲藻毒素单克隆抗体的制备及其ELISA检测方法的建立.长春:吉林大学),2011.
[11] Zhou Y,Pan F G,Li Y S,Zhang Y Y,Zhang J H,Lu S Y,Ren H Y,Liu Z S. Biosens. Bioelectron.,2009,24(8):2744-2747.
[12] Hua Y S,Cole R B. Anal. Chem.,2000,72(2):376-383.
[13] Nozawa A,Tsuji K,Ishida H. Toxicon,2003,42(1):91-103.
[14] Ishida H,Nozawa A,Hamano H,Naoki H,Fujita T,Kaspar H F,Tsuji K. Tetrahedron Lett.,2004,45(1):29-33.
[15] Shin C,Hwang J Y,Yoon J H,Kim S H,Kang G J. Food Control,2018,91(9):365-371.
[2] Bie C C,Li F M,Li Y Y,Wang Z Y. Environ. Sci.(别聪聪,李锋民,李媛媛,王震宇.环境科学),2012,33(2):442-447.
[3] Li F M,Zhao W,Li Y Y,Tian Z J,Wang Z Y. Environ. Sci.(李锋民,赵薇,李媛媛,田志佳,王震宇.环境科学),2012,33(1):233-238.
[4] Li Y Y. The Effect of Allelochemicals on Brevetoxin B Content of Gymnodinium breve. Qingdao:Ocean University of China(李媛媛.化感物质对短裸甲藻产生毒素的影响.青岛:中国海洋大学),2012.
[5] Zhang J H,Zhou Y,Zhang Y Y,Li Z H,Wang X R,Liu J Q. Animal Husbandry&Veterinary Medicine(张俊辉,周玉,张媛媛,李兆辉,王心蕊,刘静秋.畜牧与兽医),2009,41(10):5-7.
[6] Tian R Y. Screening and Preliminary Application of a ss DNA Aptamer for Detection of Brevetoxin-2. Changchun:JilinUniversity(田瑞云.短裸甲藻毒素-2适配子的制备及在检测中的初步应用.长春:吉林大学),2014.
[7] GB/T 5009. 261-2016. Determination of Neurotoxic Shellfish Poisons in Shellfish. National Standards of the People's Re-public of China(贝类中神经性贝类毒素的测定.中华人民共和国国家标准).
[8] Dickey R W,Plakas S M,Jester E L E,EI Said K R. Harmful Algae,2002,1(3):300-302.
[9] Wu H Y,Guo M M,Tan Z J,Li Z X,Zhai Y X. Chinese Fishery Quality and Standards(吴海燕,郭萌萌,谭志军,李兆新,翟毓秀.中国渔业质量与标准),2011,1(1):31-39.
[10] Wang L Q. Preparation of Monoclonal Antibody Against Brevetoxin and Establishment of ELISA Detected Method. Chang-chun:Jilin University(王里奇.短裸甲藻毒素单克隆抗体的制备及其ELISA检测方法的建立.长春:吉林大学),2011.
[11] Zhou Y,Pan F G,Li Y S,Zhang Y Y,Zhang J H,Lu S Y,Ren H Y,Liu Z S. Biosens. Bioelectron.,2009,24(8):2744-2747.
[12] Hua Y S,Cole R B. Anal. Chem.,2000,72(2):376-383.
[13] Nozawa A,Tsuji K,Ishida H. Toxicon,2003,42(1):91-103.
[14] Ishida H,Nozawa A,Hamano H,Naoki H,Fujita T,Kaspar H F,Tsuji K. Tetrahedron Lett.,2004,45(1):29-33.
[15] Shin C,Hwang J Y,Yoon J H,Kim S H,Kang G J. Food Control,2018,91(9):365-371.
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