超高效液相色谱-串联质谱法测定织纹螺中腹泻性贝类毒素的实验研究

Determination of three diarrheic shellfish toxins in Nassariidae by ultra performance liquid chromatography-mass/mass spectrometry

目的建立织纹螺组织中3种腹泻性贝类毒素（diarrhetic shellfish toxins,DSTs）：大田软海绵酸（okadaic acid,OA）;鳍藻毒素-1（dinophysistoxin-1,DTX-1）;鳍藻毒素-2（dinophysistoxin-2,DTX-2）的超高效液相色谱-串联质谱（UPLC-MS/MS）检测方法。方法样本经甲醇∶水（4∶1,V/V）溶液提取,经正己烷脱脂和C18固相萃取柱净化,经ACQUITY UPLC BEH C18（1.7μm,2.1mm×100mm）色谱柱分离,乙腈-甲酸溶液（10 mmoL/L）为流动相,流速为0.2 ml/min,柱温为30℃,负离子扫描,在多反应监测（MRM）模式下进行定性与定量分析。结果 OA、DTX-1、DTX-2在（0.01-0.20）mg/L范围内线性关系良好,3个添加水平的平均回收率为80.3%-93.6%,相对标准偏差（RSD）为2.13%-4.23%。其定量限（LOQ）为6.93μg/kg、3.3μg/kg、4.95μg/kg。结论利用此方法发现福建织纹螺中含有腹泻性贝类毒素,方法灵敏度高,操作简便,适用于实际样本的分析检测。

Objective To determine three diarrheic shellfish toxins（DSTs）by using ultra performance liquid chromatography-mass/mass spectrometry（UPLC-MS/MS）in Nassariidae. Methods Nassariidae samples were collected and extracted by methanol and water（4∶1,V/V）.Nassariidae extraction solution was processed with n-hexane and then purified by C18solid-phase extraction column.The three DSTs including Okadaic acid（OA）,Dinophysistoxin-1（DTX-1）and Dinophysistoxin-2（DTX-2）were separated on a reversed phase ACQUITY UPLC BEH C18（1.7μm,2.1mm×100mm）column using acetonitrile-10mmoL/L formic acid in water as mobile phase,with a flow rate at 0.2ml/min and a column temperature of 30℃and then were qualified and quantified by ultra performance liquid chromatography-mass/mass spectrometry under multiple reaction monitoring（MRM）. Results The method showed a good linearity of three DSTs in the range of0.01-0.2mg/L.The average recoveries ranged from 80.3%to 93.6%at three spiked concentrations with the relative standard deviation of 2.13%to 4.23%.The quantification limits（LOQ）（S/N≥10）of the method for three DSTs were 6.93μg/kg,3.3μg/kg and 4.95μg/kg,respectively. Conclusions By the established method,DSTs are detected in Nassariidae in Fujian,the method is highly sensitive and simple,and is suitable for determination of DST in many kinds of samples.