超高效液相色谱-串联质谱法测定双壳贝类中9种腹泻性贝类毒素  

Determination of nine diarrhetic shellfish poisonings in bivalve molluscs by ultra performance liquid chromatography-mass/mass spectrometry

作　　者：卢嘉丽 郑仁锦[1] 管秋美 黄宏南[1,2] 林守二 杨艳[1] 

机构地区：[1]福建省疾病预防控制中心福建省人兽共患病研究重点实验室,福建福州350001 [2]福建医科大学药学院,福建福州350004

出　　处：《中国食品卫生杂志》2017年第6期690-694,共5页Chinese Journal of Food Hygiene

基　　金：福建省自然科学基金资助项目(2015J01296)

摘　　要：目的建立超高效液相色谱-串联质谱(UPLC-MS/MS)检测方法,测定双壳贝类样品中9种腹泻性贝类毒素(DSP)。方法样品经100%甲醇提取过滤后,经ACQUITY UPLC BEH C_(18)(2.1 mm×100 mm,1.7μm)色谱柱分离,乙腈-0.01%氨水为流动相,流速为0.2 ml/min,柱温为30℃,采用电喷雾电离和正负离子分段扫描,在多反应监测(MRM)模式下测定游离态的毒素(原多甲藻酸1、原多甲藻酸2、原多甲藻酸3、大环内酯类扇贝毒素2、软海绵酸、鳍藻毒素1、鳍藻毒素2、虾夷扇贝毒素和类虾夷扇贝毒素)。当软海绵酸、鳍藻毒素1或鳍藻毒素2任一种或多种毒素呈阳性时,须采用碱水解将酯化的软海绵酸、鳍藻毒素1和鳍藻毒素2转化为游离态形式后,用UPLCMS/MS测定其水解态毒素含量,外标法定量。结果 9种DSP在3个添加水平的平均回收率为85.9%~95.3%,相对标准偏差(RSD)为2.76%~4.18%,检出限(LOD)为4.4~9.9μg/kg,定量限(LOQ)为14.7~33.0μg/kg。结论该方法操作简便、灵敏度高,适用于DSP的日常检测。Objectice Determination method of nine diarrhea shellfish poisons by ultra performance liquid chromatography-mass/mass spectrometry（ UPLC-MS/MS） in bivalve molluscs was established. Methods The samples were extracted by methanol and separated on a reversed phase ACQUITY UPLC BEH C_（18）（ 2. 1 mm × 100 mm,1. 7 μm）column with acetonitrile-0. 01 % ammonium hydroxide as the mobile phase,with a flow rate at 0. 2 ml/min and the column temperature of 30 ℃. The free toxins（ AZA1, AZA2, AZA3, PTX2-b, OA, DTX1, DTX2, YTX and h YTX） in the samples were detected under positive and negative multiple reactions monitoring（ MRM） mode through polarity switching between time segments. When any one or more toxins of OA,DTX1 or DTX2 were positive in the sample,the esterified OA,DTX1 and DTX2 were converted to the free form by alkaline hydrolysis and their hydrolyzed toxins were determined by UPLC-MS/MS and quantified by external standard method. Results The average recoveries ranged from 85. 9 % to95. 3 % at three spiked concentrations with the relative standard deviation（ RSD） of 2. 76 % to 4. 18 %. The detection limit（ LOD） ranged from 4. 4 to 9. 9 μg/kg. The quantification limit（ LOQ） ranged from 14. 7 to 33. 0 μg/kg. Conclusion The method was highly sensitive and simple, and was suitable for daily detection of diarrhea shellfish poisons in bivalve molluscs.

关 键 词：超高效液相色谱-串联质谱法 双壳贝类 腹泻性贝类毒素 食品污染物 食品安全 检测方法 

分 类 号：O657.63[理学—分析化学] TS254.7[理学—化学]