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Abstract：
In this paper, an LC-MS/MS method for the simultaneous identification and quantification of cyanotoxins with hydrophilic and lipophilic properties in edible bivalves is presented. The method includes 17 cyanotoxins comprising 13 microcystins (MCs), nodularin (NOD), anatoxin-a (ATX-a), homoanatoxin (h-ATX) and cylindrospermopsin (CYN). A benefit to the presented method is the possibility for the MS detection of MC-LR-[Dha7] and MC-LR-[Asp3] as separately identified and MS-resolved MRM signals, two congeners which were earlier detected together. The performance of the method was evaluated by in-house validation using spiked mussel samples in the quantification range of 3.12-200 µg/kg. The method was found to be linear over the full calibration range for all included cyanotoxins except CYN for which a quadratic regression was used. The method showed limitations for MC-LF (R2 = 0.94), MC-LA (R2 ≤ 0.98) and MC-LW (R2 ≤ 0.98). The recoveries for ATX-a, h-ATX, CYN, NOD, MC-LF and MC-LW were lower than desired (<70%), but stable. Despite the given limitations, the validation results showed that the method was specific and robust for the investigated parameters. The results demonstrate the suitability of the method to be applied as a reliable monitoring tool for the presented group of cyanotoxins, as well as highlight the compromises that need to be included if multi-toxin methods are to be used for the analysis of cyanotoxins with a broader range of chemical properties. Furthermore, the method was used to analyze 13 samples of mussels (Mytilus edulis) and oysters (Magallana gigas) collected in the 2020-2022 summers along the coast of Bohuslän (Sweden). A complementary qualitative analysis for the presence of cyanotoxins in phytoplankton samples collected from marine waters around southern Sweden was performed with the method. Nodularin was identified in all samples and quantified in bivalve samples in the range of 7-397 µg/kg. Toxins produced by cyanobacteria are not included in the European Union regulatory monitoring of bivalves; thus, the results presented in this study can be useful in providing the basis for future work including cyanotoxins within the frame of regulatory monitoring to increase seafood safety.
摘要：
在本文中,提出了一种LC-MS/MS方法，用于同时鉴定和定量可食用双壳类动物中具有亲水和亲脂特性的氰基毒素。该方法包括17种氰基毒素，包括13种微囊藻毒素（MC），nodularin（NOD），Anatoxin-a(ATX-a),高抗毒素（h-ATX）和圆柱精子素（CYN）。所提出的方法的一个好处是MS检测MC-LR-[Dha7]和MC-LR-[Asp3]作为单独识别和MS分辨的MRM信号的可能性。之前一起发现的两个同源物。通过在3.12-200µg/kg的定量范围内使用加标的贻贝样品进行内部验证来评估该方法的性能。除了使用二次回归的CYN外，发现该方法在所有包含的氰基毒素的整个校准范围内都是线性的。该方法显示了MC-LF的局限性(R2=0.94)，MC-LA(R2≤0.98)和MC-LW(R2≤0.98)。ATX-a的回收，h-ATX,CYN,点头,MC-LF和MC-LW低于预期(<70%)，但稳定。尽管存在一定的局限性,验证结果表明，该方法对所研究的参数具有特异性和鲁棒性。结果表明，该方法适合用作所提供的氰基毒素组的可靠监测工具，以及强调如果要使用多毒素方法来分析具有更广泛化学性质的氰基毒素，则需要包括的妥协。此外，该方法用于分析在2020-2022年夏季沿Bohuslän（瑞典）海岸收集的13种贻贝（Mytilusedulis）和牡蛎（Magallanagigas）样品。使用该方法对从瑞典南部海域收集的浮游植物样品中氰基毒素的存在进行了补充定性分析。在所有样品中都鉴定出Nodularin，并在7-397µg/kg范围内的双壳类样品中进行了定量。蓝细菌产生的毒素不包括在欧盟对双壳类动物的监管监测中；因此，本研究的结果有助于为未来的工作提供基础，包括在监管监测框架内的氰基毒素，以提高海鲜安全。
