Food Science and Nutrition Department, XaRTA (Catalonian Reference Network on Food Technology), Campus de l'Alimentació de Torribera, University of Barcelona, Av/ Prat de la Riba, 171, 08921 Santa Coloma de Gramenet, Spain; Mass Spectrometry Laboratory/Organic Pollutants, IDAEA-CSIC, c/ JordiGirona, 18-26, 08034 Barcelona, Spain.
Mass Spectrometry Laboratory/Organic Pollutants, IDAEA-CSIC, c/ JordiGirona, 18-26, 08034 Barcelona, Spain.
J Chromatogr A. 2014 Feb 7;1328:16-25. doi: 10.1016/j.chroma.2013.12.071. Epub 2013 Dec 31.
A multitoxin method has been developed for quantification and confirmation of lipophilic marine biotoxins in mussels by liquid chromatography coupled to high resolution mass spectrometry (HRMS), using an Orbitrap-Exactive HCD mass spectrometer. Okadaic acid (OA), yessotoxin, azaspiracid-1, gymnodimine, 13-desmethyl spirolide C, pectenotoxin-2 and Brevetoxin B were analyzed as representative compounds of each lipophilic toxin group. HRMS identification and confirmation criteria were established. Fragment and isotope ions and ion ratios were studied and evaluated for confirmation purpose. In depth characterization of full scan and fragmentation spectrum of the main toxins were carried out. Accuracy (trueness and precision), linearity, calibration curve check, limit of quantification (LOQ) and specificity were the parameters established for the method validation. The validation was performed at 0.5 times the current European Union permitted levels. The method performed very well for the parameters investigated. The trueness, expressed as recovery, ranged from 80% to 94%, the precision, expressed as intralaboratory reproducibility, ranged from 5% to 22% and the LOQs range from 0.9 to 4.8pg on column. Uncertainty of the method was also estimated for OA, using a certified reference material. A top-down approach considering two main contributions: those arising from the trueness studies and those coming from the precision's determination, was used. An overall expanded uncertainty of 38% was obtained.
已开发出一种多毒素方法，用于通过液相色谱-高分辨质谱（HRMS）联用技术，使用轨道阱-Exactive HCD 质谱仪，对贝类中的脂溶性海洋生物毒素进行定量和确证。将冈田酸（OA）、鳍藻毒素-1、短裸甲藻素-1、裸甲藻毒素-G、13-去甲基短裸甲藻素 C、扇贝毒素-2 和膝沟藻毒素 B 作为每个脂溶性毒素组的代表性化合物进行分析。建立了 HRMS 鉴定和确证标准。研究并评估了碎片和同位素离子以及离子比，以用于确证目的。对主要毒素的全扫描和碎片谱进行了深入的特征描述。准确度（准确性和精密度）、线性、校准曲线检查、定量限（LOQ）和特异性是为方法验证建立的参数。验证在当前欧盟允许水平的 0.5 倍进行。该方法在研究的参数方面表现非常出色。真实度（表示为回收率）在 80%至 94%之间，精密度（表示为实验室内部重现性）在 5%至 22%之间，LOQ 在 0.9 至 4.8pg 之间。还使用认证参考物质对 OA 进行了方法不确定度的评估。采用自上而下的方法，考虑了两个主要贡献：来自真实度研究的贡献和来自精密度确定的贡献。得到了 38%的总扩展不确定度。
