超高压液相色谱-高分辨质谱快速筛查和确证食用贝类中多种原多甲藻酸贝类毒素

Rapid Profiling and Confirmation of Azaspiracids in Edible Shellfishes by Ultra High Performance Liquid Chromatography-High Resolution Mass Spectrometry

建立超高压液相色谱-高分辨质谱快速筛查和确证贻贝、牡蛎、蚌类、扇贝等食用贝类及其制品中3种天然形式的原多甲藻酸贝类毒素的检测方法。样品中的原多甲藻酸采用乙腈-水体系均质提取，应用改进型QuEChERS技术净化，在乙腈-水（含5mmol／L醋酸铵和0．1％甲酸）体系经AcquityHSST3柱（150mm×2．1mm，1.8μm）梯度洗脱，实现了3种原多甲藻酸贝类毒素的基线分离。该方法基于电喷雾正离子模式，采用高分辨质谱一级全扫描和数据依赖扫描，对食用贝类及其制品样品中的原多甲藻酸贝类毒素进行检测。3种贝类毒素的定量限均为10μg／kg（RSN〉10）；在10-500μg／L范围内线性关系良好，相关系数（R^2）均大于0．99。应用该方法对国内外多个地区的贝类产品进行了筛查及确证，其中部分样品检出原多甲藻酸贝类毒素。该方法灵敏度高，重复性好，操作简便、快捷，适用于食用贝类及其制品中多种原多甲藻酸贝类毒素的筛查分析。

An ultra high performance liquid chromatography-high resolution mass spectrometric method （UHPLC-LTQ/ Orbitrap） was developed for the rapid profiling and confirmation of 3 natural azaspiracids （AZAs） （AZA-1, AZA-2 and AZA-3） in edible shellfishes including mussels, oysters, clams and scallops as well as related products. Samples were homogeneously extracted with acetonitrile-water. The diluted supematants were then purified with a modified QuEChERS method. The separation was performed on an Acquity HSS T3 column （150 mm ×2.1mm,1.8μm） by gradient elution with acetonitrile in water containing 5 mmol/L ammonium acetate and 0.1% formic acid. The high-resolution mass spectrometry was carried out by means of electrospray ionization in positive ion mode （ESI＋）. The AZAs were detected by high-resolution MS under full scan and data-dependent scan modes, respectively. The limits of quantification （LOQ, RSN〉 10） were 10 μg/kg for all the 3 AZAs. The calibration curves showed good linearity within the concentration range of 10 μg/L to 500μg/L with correlation coefficients （R2） more than 0.99. Shellfish samples from home and abroad were profiled and confirmed by the established method. AZAs were found in some samples. Therefore this method is easy, sensitive, reproducible and efficient, and can be applied to profile and confirm AZAs in edible shellfishes and related products.