An analytical method using liquid chromatography tandem mass spectrometry has been developed for the determination of neurotoxic shellfish poisoning (NSP) toxins (brevetoxins) in shellfish. The tested parameters included extraction solvent, clean-up method, mobile phase and mass spectrometric detection parameters including multiple reaction monitoring (MRM) conditions. The toxins were extracted with 80% methanol in water and cleaned up using Strata-X SPE cartridges. Chromatographic separation of brevetoxins (PbTx-1, PbTx-2 and PbTx-3) was carried out on a C18 column (100 mm × 2.1 mm, 2.6 μm) with gradient elution. The developed method was validated according to the requirements of SANTE 11945/2015. The average recoveries of the brevetoxins ranged from 75.9% to 105.8%, the intra-day precisions were in the range of 0.4–8.6%, the inter-day precisions were in the range of 0.7–6.3% and the limits of quantification (LOQs) were 25 μg/kg for each toxin. The validated method was successfully applied to real shellfish samples, including mussel, oyster and blood cockle. Overall, the method has shown excellent sensitivity and reproducibility, and it could be used as a reference method for the monitoring of individual brevetoxins in shellfish samples.

已经开发了一种使用液相色谱串联质谱法的分析方法，用于测定贝类中的神经毒性贝类中毒（NSP）毒素（短毒素）。测试的参数包括萃取溶剂，净化方法，流动相和质谱检测参数，其中包括多反应监测（MRM）条件。用80％的甲醇水溶液提取毒素，并使用Strata-X SPE柱进行净化。在C18色谱柱（100 mm×2.1 mm，2.6μm）上用梯度洗脱法对短毒素（PbTx-1，PbTx-2和PbTx-3）进行色谱分离。根据SANTE 11945/2015的要求对开发的方法进行了验证。brevetoxins的平均回收率在75.9％至105.8％之间，日内精确度在0.4-8.6％的范围内，日间精度在0.7–6.3％的范围内，每种毒素的定量限（LOQ）为25μg/ kg。经过验证的方法已成功应用于包括贝类，牡蛎和血蛤在内的真实贝类样品。总体而言，该方法显示出极好的灵敏度和可重复性，可作为监测贝类样品中单个短毒素的参考方法。