Abstract Analysis of marine biotoxins in high-lipid bivalves has several limitations due to the coexistence of lipid matrices, low biotoxin concentrations, and the different physiochemical properties of the various biotoxins. In this study, a sensitive and reliable method to simultaneously determine six diarrhetic shellfish poisoning (DSP) toxins (five acidic and one neutral biotoxins) in oysters and mussels was developed based on UHPLC-MS/MS-multiple ion reaction monitoring (MRM) using time segment polarity switching. For effective sample pretreatment, three protocols including syringe filtration, Strata™-X solid phase extraction (SPE), and freezing lipid filtration combined with SPE (FLF + SPE) were evaluated, and FLF + SPE method was shown to be the most effective with high lipid removal efficiency. Aging effects of the UHPLC mobile phase on MS detection sensitivity and retention time shift of DSP toxins were carefully examined. The developed method provided good recoveries ranging from 81.0 to 119.6%, showing no significant matrix effect and assay was linear with coefficients of determination above R 2 > 0.99 for all analytes. Validation tests using a certified reference material (CRM) and spiking DSP toxins (OA and DTX-1 at 1.07 μg/g, and DTX-2 at 0.86 μg/g) showed this method to be acceptable, with a relative standard deviation of

中文翻译：

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UHPLC-ESI-MS/MS 在时间段极性切换模式下可靠定量高脂双壳类动物中痕量腹泻性贝类中毒毒素：三种提取方法的比较

摘要 由于脂质基质共存、生物毒素浓度低以及各种生物毒素的不同理化性质，高脂双壳类海洋生物毒素的分析存在一些局限性。在本研究中，基于 UHPLC-MS/MS-多离子反应监测 (MRM)，开发了一种灵敏可靠的方法，可同时测定牡蛎和贻贝中的六种腹泻性贝类中毒 (DSP) 毒素（五种酸性生物毒素和一种中性生物毒素）。时间段极性切换。对于有效的样品预处理，评估了三种方案，包括注射器过滤、Strata™-X 固相萃取 (SPE) 和冷冻脂质过滤结合 SPE (FLF + SPE)，结果表明 FLF + SPE 方法最有效。去脂效率高。仔细研究了 UHPLC 流动相对 MS 检测灵敏度和 DSP 毒素保留时间偏移的老化影响。所开发的方法提供了 81.0% 到 119.6% 的良好回收率，没有显示出显着的基质效应，并且所有分析物的测定系数高于 R 2 > 0.99 时呈线性。使用有证参考物质 (CRM) 和加标 DSP 毒素（OA 和 DTX-1 为 1.07 μg/g，DTX-2 为 0.86 μg/g）的验证测试表明该方法是可以接受的，相对标准偏差为