液相色谱-串联质谱法测定双壳类水产品中原多甲藻酸类贝类毒素

Determination of Azaspiracids Shellfish Poison in Bivalve Aquatic Product by Liquid Chromatography Tandem Mass Spectrometry

建立了双壳类水产品中原多甲藻酸贝类毒素Azaspiracid 1(AZA1)、Azaspiracid 2(AZA2)和Azaspir-acid 3(AZA3)的液相色谱-电喷雾串联质谱检测方法。选用具有基质代表性的扇贝、牡蛎和杂色蛤为研究对象,样品经80%甲醇-水混合溶液提取,正己烷脱脂、氯仿反提萃取并旋转蒸发浓缩后,MAX混合型阴离子交换反相吸附固相萃取柱净化,Luna C18色谱柱(150 mm×2.0 mm,5μm,Phenomenex公司)反相液相色谱法分离,乙腈和0.1%甲酸溶液组成的流动相梯度洗脱,正离子扫描,多反应监测(MRM)模式下电喷雾串联质谱法测定和确证,外标法定量。结果表明,3种原多甲藻酸类贝类毒素均在4 min内出峰,检出限均为1.5μg/kg,在0.3～30μg/L范围内线性良好,平均回收率大于80%,相对标准偏差(RSD)小于12%(n=6)。

A method was established for the simultaneous determination of three main kinds of azaspiracids （AZAs） shellfish poison, such as azaspiraeid 1 （AZA1）, azaspiracid 2 （AZA2）, azaspiracid 3 （AZA3） in bivalve aquatic product by liquid chromatography tandem mass spectrometry. The matrix representative shellfish including scallop, oyster and clam were extracted with 80% methanol-water solution, defatted by hexane, reextracted by chloroform subsequently, and then enriched by rotary evaporation before purified by solid phase extraction on a MAX solid phase extraction cartridge which contains a kind of mixed-mode sorbent with reversed-phase and anion-exchange functionalities. The analytes were isolated by reversed phase high performance liquid chromatography on a Luna C18 column （150 mm x 2.0 mm, 5 ~m） and gradient eluted with acetonitrile-0.1% formic acid at a flow rate of 1.0 mL/min. AZAs were determined with the multiple reaction monitoring （MRM） mode by tandem mass spectrometry with electrospray detector through positive ion scanning. Quantitative analysis method was external standard method. The experimental results indicated that the retention time of AZAs was all within 4 rain and the limit of quantification （ LOQ ） of them were all 1.5 ~g/kg respectively, the calibration curves were linear in the range from 0.3 ~g/L to 30 p.g/L, the average recoveries at spiked concentration of 1.5, standard deviations （RSDs） were less than 12% （ 60and 120 Ixg/kg were more than 80% and the relative n=6）.