超高效液相色谱-串联质谱法快速测定水产加工食品中的河豚毒素
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摘要
建立了快速测定水产加工食品中河豚毒素(tetrodotoxin,TTX)的超高效液相色谱-串联质谱(UPLC-MS/MS)分析方法。样品经0.1%乙酸水溶液提取,C18萃取剂分散固相萃取净化,超滤管过滤后进行分析。目标物TTX采用TSK-gel Amide-80亲水色谱柱进行分离,以电喷雾离子源、多反应监测(MRM)正离子模式检测,基质标准曲线校正,外标法定量。结果表明,TTX在2~200 ng/mL范围内线性良好(r2>0.999),方法的定量限为10μg/kg,加标回收率为71.2%~102%,相对标准偏差(RSD)为5.23%~12.4%。方法操作简便,准确度和精密度良好,可用于烤鱼片、鱼干等干制水产加工食品中河豚毒素的测定,具有较好的应用价值。
A method for the rapid determination of tetrodotoxin(TTX) in aquatic food using ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS / MS) has been established.TTX was extracted from sample by 0.1% acetic acid-water,and then purified by dispersive solid-phase extraction with C 18 extraction sorbent and filtered through a Mr 10000 cut-off ultrafree microcentrifuge filter.The separation of TTX was carried out on a column of TSK-gel Amide-80,and detected by positive electrospray ionization tandem mass spectrometry in the MRM mode.The matrix-matched external standard calibration curves were used for quantitative analysis.The calibration curves were linear in the range of 2-200 ng / mL for TTX,with quantification limit of 10 μg / kg.The average recoveries were between 71.2% and 102% with relative standard deviations(RSD) ranging from 5.23% to 12.4%.The proposed method was simple,rapid,accurate and applicable for the determination of TTX in aquatic food.
A method for the rapid determination of tetrodotoxin(TTX) in aquatic food using ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS / MS) has been established.TTX was extracted from sample by 0.1% acetic acid-water,and then purified by dispersive solid-phase extraction with C 18 extraction sorbent and filtered through a Mr 10000 cut-off ultrafree microcentrifuge filter.The separation of TTX was carried out on a column of TSK-gel Amide-80,and detected by positive electrospray ionization tandem mass spectrometry in the MRM mode.The matrix-matched external standard calibration curves were used for quantitative analysis.The calibration curves were linear in the range of 2-200 ng / mL for TTX,with quantification limit of 10 μg / kg.The average recoveries were between 71.2% and 102% with relative standard deviations(RSD) ranging from 5.23% to 12.4%.The proposed method was simple,rapid,accurate and applicable for the determination of TTX in aquatic food.
引文
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[16]舒静,李柏林,欧杰.离子色谱法检测酒曲发酵液中的河豚毒素[J].色谱,2011,29(2):187-190.
[17]金玉娥,马佳鸣,熊丽蓓,等.HPLC-MS法测定鲜鱼中河豚毒素[J].中国司法鉴定,2012(2):22-25.
[2]张秀尧,蔡欣欣.亲水液相色谱三重四极杆质谱联用法快速检测尿液和血浆中河豚毒素[J].分析化学,2009,12(12):1829-1833.
[3]李世敏.应用营养与食品卫生管理[M].北京:中国农业出版社,2002:196-208.
[4]苏文,薛际深,李维刚.河豚毒素的检测分析[J].防化研究,1992(1):54-64.
[5]殷芹,刘岩,宫庆礼.河豚毒素的理论及应用研究进展[J].海洋通报,2008,27(6):95-100.
[6]马群飞,林升清,王明.烤鱼片干河豚毒素中毒事件的追踪调查[J].中国卫生监督杂志,2001,8(4):145-147.
[7]周晓翠,谢光洪,刘国文,等.河豚毒素检测方法的研究进展[J].中国畜牧兽医,2008,35(7):43-47.
[8]崔建洲,申雪艳.高效液相色谱-紫外/荧光检测方法测定河豚毒素[J].色谱,2006,24(3):317.
[9]宫庆礼,黄海洲.河豚毒素的色谱检测[J].水产科学,2001,20(6):31-33.
[10]戴月,陶宁萍,刘源,等.基质固相分散高效液相色谱法检测河豚毒素[J].光谱实验室,2012,29(3):1601-1604.
[11]骆和东,贾玉珠,朱宝平.固相萃取-超过滤-液相色谱/质谱联用法测定织纹螺中的河豚毒素[J].色谱,2007,25(6):917-921.
[12]李爱峰,于仁成.液相色谱-电喷雾离子阱质谱联用分析河豚毒素[J].分析化学,2007,35(3):397-400.
[13]Nakagawa T,Jang J,Yotsu-Yamashita M.Hydrophilic interaction liquid chromatography-electrospray ionization mass spectrometry of tetrodotoxin and its analogs[J].Anal Biochem,2006(352):142-144.
[14]Hayashida M,Hayakawa H,Wada K,et a1.A column-switching LC/MS/ESI method for detecting tetrodotoxin and Aconiturn alkaloids in serum[J].Leg Med,2003(5):101-104.
[15]崔建升,陈婧.微生物传感器测定河豚毒素研究[J].海洋环境科学,2009,28(6):726-728.
[16]舒静,李柏林,欧杰.离子色谱法检测酒曲发酵液中的河豚毒素[J].色谱,2011,29(2):187-190.
[17]金玉娥,马佳鸣,熊丽蓓,等.HPLC-MS法测定鲜鱼中河豚毒素[J].中国司法鉴定,2012(2):22-25.