液相色谱-四极杆/线性离子阱复合质谱测定双壳贝类中麻痹性贝类毒素

摘要

We developed a new method for simultaneous identification and detection of 13 paralytic shellfish toxins (PSTs) in bivalve mollusks using liquid chromatography coupled with quadrupole/linear ion trap tandem mass spectrometry (LC-QTrap/MS).Samples were extracted with acetic acid aqueous solution,and cleaned up in ENVI-Carb solid-phase extraction column.Separation of the 13 PSTs were performed on an HILIC (hydrophilic interaction liquid chromatography) with multiple reaction monitoring (MRM)-information-dependent acquisition (IDA) experiment-enhanced product ion (EPI) scan in mass spectrometry acquisition.The calibration curves are linear well with correlation coefficient over 0.99.The overall detection limit of the method is 62.0μg for STX eq/kg;specifically,the limit for STX,dcSTX,NEO,dcNEO,and GTX5 was 10.0μg/kg;12.0μg/kg for GTX1;4μg/kg for GTX3,GTX4,C2 and dcGTX3,and 131μg/kg for C1 and dcGTX2.The average spiked recoveries for 13PSTs were (79.6±10.4)％-(98.8±6.54)％.This method could reduce shellfish matrix suppression effect,simplify pre-treatment,reduce the times of dilution,and increase the sensitivity,during which isomers could be separated from baseline.The proposed method therefore is recommended to identify and detect the 13PSTs in bivalve mollusks.%采用液相色谱-四极杆/线性离子阱复合质谱,建立了双壳贝类中13种麻痹性贝类毒素的定性确证和定量分析方法.样品经乙酸水溶液提取,石墨化碳黑固相萃取净化,亲水性液相色谱柱(hydrophilic interaction liquid chromatography,HILIC)分离;质谱采集使用独有的多反应监测-信息依赖性采集-增强子离子扫描模式.13种目标物线性范围相关系数不低于0.99,检出限为62.0μg STX eq/kg,其中石房蛤毒素(Saxitoxin,STX)、新石房蛤毒素(Neosaxitoxin,NEO)、脱氨甲酰基石房蛤毒素(Decarbamoylsaxitoxin,dcSTX)、脱氨甲酰基新石房蛤毒素(Decarbaoylneosaxitoxin,dcNEO)以及N-磺酰氨甲酰基类毒素5(Gonyautoxin-5,GTX5)为10.0μg/kg,膝沟藻毒素1&2(Gonyautxins-1&-2,GTX1&2)为12.0μg/kg,膝沟藻毒素3&4 (Gonyautxins-3&-4,GTX3&4)、N-磺酰氨甲酰基类毒素2(N-Sulfocarbamoylgonyautoxin-2,C2)和脱氨甲酰基膝沟藻毒素3(Decarbamoylgonyautoxins-3,dcGTX3)为4上g/kg,N-磺酰氨甲酰基类毒素1(N-Sulfocarbamoylgonyautoxin-1,C1)和脱氨甲酰基膝沟藻毒2(Decarbamoylgonyautoxins-2,dcGTX2)为13μ g/kg.基质加标平均回收率为(79.6±10.4)％—(98.8±6.54)％.该方法能够有效降低贝类基质抑制效应,简化前处理过程并通过减少样品稀释倍数来显著提高方法灵敏度,使同分异构体达到基线分离,适用于双壳贝类中麻痹性贝类毒素的监控分析.