对虾体内4种真菌毒素超声萃取LC-MS/MS检测及其蓄积分布特征Ultrasonic-assisted Extraction of Four Mycotoxins in Shrimp Simultaneously Determined by LC-MS/MS and the Distribution Characteristics of Mycotoxin Accumulation in Shrimp Tissues
王雅玲,邓义佳,王润东,孙力军,邓旗,王小博,廖建萌
摘要(Abstract):
【目的】建立超声辅助萃取高效液相色谱-串联质谱法(Liquid chromatography-tandem mass spectrometry,LC-MS/MS)同步检测对虾各组织及血液中黄曲霉毒素(AflatoxinB1,AFB1)、T-2毒素(T-2)、赭曲霉毒素A(Ochratoxin A,OTA)与脱氧雪腐镰刀菌烯醇(Deoxynivalenol,DON)。【方法】取对虾肌肉、头胸部、虾壳、肝胰腺、肠道和血液样品,采用乙腈/水(体积比85∶15)为提取溶剂,20℃超声辅助萃取60 min,正己烷脱脂,以MycoSpinTM 400柱净化,采用液相色谱-串联质谱法进行检测,用Hypersil Gold-C18色谱柱分离;以流动相甲醇和5 mmol/L的乙酸铵(含0.1%甲酸)进行梯度洗脱,流速为0.25 mL/min;采用正负离子扫描模式,外标法定量。【结果】4种真菌毒素在各自线性范围内线性良好,相关系数(r~2)均大于0.99,定量限为0.3~3.0μg/kg,回收率为75.0%~105.4%,相对标准偏差(Relative standard deviation, RSD)为2.1%~14.5%,通过超声辅助萃取条件优化,显著提高了对虾组织中真菌毒素的提取率。【结论】LC-MS/MS方法前处理简单、检测速度快、净化效果好、结果准确、可靠,适合对虾各组织中多种真菌毒素的快速检测。真菌毒素在对虾不同组织中的蓄积特征,AFB1、T-2、OTA和DON均主要蓄积于对虾肠道与血液中,蓄积率分别达到43%~52%和31%~45%,头部、肌肉、肝胰腺和外壳蓄积率均低于10%,并非真菌毒素残留的靶向组织。
关键词(KeyWords): 真菌毒素;液相色谱-串联质谱(LC-MS/MS);超声辅助萃取;凡纳滨对虾;蓄积特征
基金项目(Foundation): 国家自然科学基金面上项目(31871898);; 广东省现代农业产业技术体系创新团队建设专项资金项目(2019KJ149,2019KJ151);; 广东海洋大学科研启动经费资助项目(R19050)
作者(Author): 王雅玲,邓义佳,王润东,孙力军,邓旗,王小博,廖建萌
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