A Feasibility Study into the Production of a Mussel Matrix Reference Material for the Cyanobacterial Toxins Microcystins and Nodularins.
A Feasibility Study into the Production of a Mussel Matrix Reference Material for the Cyanobacterial Toxins Microcystins and Nodularins.
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DOI：
10.3390/toxins15010027
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发表时间：
2022-12-30
期刊：
影响因子：
4.2
通讯作者：
Lewis AM
中科院分区：
文献类型：
--
作者：
Turner AD;Beach DG;Foss A;Samdal IA;Løvberg KLE;Waack J;Edwards C;Lawton LA;Dean KJ;Maskrey BH;Lewis AM
由某些种类的蓝藻自然产生的微囊藻毒素和节藻毒素已被发现在鱼类和贝类等水产食品中积累，对海产品消费者的健康构成风险。出于风险管理的目的，监测这类生物体中的毒素需要提供经认证的基质参考物质，以帮助方法开发、验证和常规质量保证。因此，这项研究的目标是制备一种由一系列微囊藻毒素类似物和节瘤蛋白引起的贻贝组织标准物质。通过LC-MS/MS(LC-MS/MS)确认，在材料中加入了9个目标类似物，并使用LC与非目标高分辨率质谱仪(LC-HRMS)联用检测到另外15个类似物。采用LC-MS/MS、两种不同的酶联免疫吸附试验(ELISA)和氧化裂解法定量3-甲氧基-2-甲基-4-苯基丁酸(MMPB)。使用不同方法量化的浓度之间的相关性是不同的，可能与检测交叉反应的不同以及每种方法检测结合毒素的能力不同有关。由四种独立测试方法测定的总可溶性毒素的普遍浓度为2425±575微克/千克湿重。除可自由提取的可溶物(57±9%)外，结合性毒素的平均含量为43±9%。所生产的标准物质在冰箱中储存六个月时是均匀和稳定的，没有任何生产后稳定剂应用。因此，已经制作了氰化毒素贝类标准物质，证明了为大量氰化毒素开发经认证的海产品基质标准物质的可行性，并可为氰化毒素风险监测、管理和缓解提供有价值的未来资源。
Microcystins and nodularins, produced naturally by certain species of cyanobacteria, have been found to accumulate in aquatic foodstuffs such as fish and shellfish, resulting in a risk to the health of the seafood consumer. Monitoring of toxins in such organisms for risk management purposes requires the availability of certified matrix reference materials to aid method development, validation and routine quality assurance. This study consequently targeted the preparation of a mussel tissue reference material incurred with a range of microcystin analogues and nodularins. Nine targeted analogues were incorporated into the material as confirmed through liquid chromatography with tandem mass spectrometry (LC-MS/MS), with an additional 15 analogues detected using LC coupled to non-targeted high resolution mass spectrometry (LC-HRMS). Toxins in the reference material and additional source tissues were quantified using LC-MS/MS, two different enzyme-linked immunosorbent assay (ELISA) methods and with an oxidative-cleavage method quantifying 3-methoxy-2-methyl-4-phenylbutyric acid (MMPB). Correlations between the concentrations quantified using the different methods were variable, likely relating to differences in assay cross-reactivities and differences in the abilities of each method to detect bound toxins. A consensus concentration of total soluble toxins determined from the four independent test methods was 2425 ± 575 µg/kg wet weight. A mean 43 ± 9% of bound toxins were present in addition to the freely extractable soluble form (57 ± 9%). The reference material produced was homogenous and stable when stored in the freezer for six months without any post-production stabilization applied. Consequently, a cyanotoxin shellfish reference material has been produced which demonstrates the feasibility of developing certified seafood matrix reference materials for a large range of cyanotoxins and could provide a valuable future resource for cyanotoxin risk monitoring, management and mitigation.
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