液相色谱-串联质谱法检测贝类产品中的原多甲藻酸贝类毒素

Determination of azaspiracid-1 in shellfishes by liquid chromatography with tandem mass spectrometry

建立了一种贝类组织中原多甲藻酸(azaspiracid,AZA)贝类毒素主要成分AZA1的高效液相色谱-串联质谱检测方法。本方法采用甲醇-水(80:20,v/v)溶液对贝类组织中AZA1进行提取,并用MAX阴离子交换固相萃取(SPE)柱富集净化,使用AtlantisdC18(150mm×4.6mm,5.0μm)色谱柱分离,以含有50mmol/L甲酸和2mmol/L甲酸铵的乙腈-水溶液(80:20,v/v)为流动相进行等度洗脱,质谱采用选择反应监测(SRM)模式。AZA1在5min内获得完全分离,且在48.85～2442ng/L范围内线性良好,相关系数为0.9981。该方法检出限(S/N=3)为11.00pg/g,添加水平为36.64、73.27、146.54pg/g时的平均回收率为75.8%～82.5%(n=6),相对标准偏差小于10%。利用该方法对采自大连、青岛、广州水产品市场上的112个贝类样品进行了分析,发现采自大连和广州的部分贝类样品中含有AZA1。结果表明,该方法具有简单、快速、灵敏度高等特点,能充分满足贝类中AZA1检测的要求。

A liquid chromatography-tandem mass spectrometry （LC-MS/MS） method for the determination of azaspiracid-1 （AZA1） in shellfishes was described.After being extracted using methanol and water （80∶20,v/v）,the extract was cleaned-up by solid phase extraction （SPE） of MAX column,then determined by using a reversed-phase high performance liquid chromatography （HPLC） isocratic program coupled with tandem mass spectrometry in selected reaction monitoring mode （SRM）.And the extract was eluted with acetonitrile-water （80∶20,v/v） on an Atlantis dC18 column （150 mm × 4.6mm,5.0 μm） with mobile phase containing 50 mmol/L formic acid and 2 mmol/L ammonium formate.The detection limit was 11.00 pg/g.The calibration curve was linear （R2=0.998 1） in the range of 48.85-2 442 ng/L.The average recoveries of the shellfish tissue extract at three spiked levels （36.64,73.27,146.54 pg/g） were from 75.8% to 82.5% （n=6）.The relative standard derivations （RSDs） were less than 10%.The 112 shellfish samples from the local markets of Dalian,Qingdao,Guangzhou were detected by the method,and AZA1 was detected in some samples from Dalian and Guangzhou.The results showed that the method is simple,rapid,sensitive and suitable for the detection of AZA1 in shellfishes.