多种产毒藻混合暴露制备麻痹性贝类毒素基体标准物质原料技术  

Technological Advances in Preparing Raw Materials of Paralytic Shellfish Toxin Matrix Standards by Mixed Exposure to Multiple Toxic Dinoflagellates

作　　者：赵辉辉 沙珍霞 王潇潇 郑关超 谭志军[2,4] 吴海燕[2] ZHAO Huihui;SHA Zhenxia;WANG Xiaoxiao;ZHENG Guanchao;TAN Zhijun;WU Haiyan(College of Life Sciences,Qingdao University,Qingdao 266071,China;Key Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality,Ministry of Agriculture and Rural Affairs,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071,China;Institute of Aquatic Biotechnology,College of Life Sciences,Qingdao University,Qingdao 266071,China;State Key Laboratory of Mariculture Biobreeding and Sustainable Goods,Yellow Sea Fisheries Research Institute,Chinese Academy of Fishery Sciences,Qingdao 266071,China)

机构地区：[1]青岛大学生命科学学院,山东青岛266071 [2]农业农村部水产品质量安全检测与评价重点实验室、中国水产科学研究院黄海水产研究所,山东青岛266071 [3]青岛大学生命科学学院水生生物技术研究院,山东青岛266071 [4]海水养殖生物育种与可持续产出全国重点实验室、中国水产科学研究院黄海水产研究所,山东青岛266071

出　　处：《渔业科学进展》2024年第5期213-223,共11页Progress in Fishery Sciences

基　　金：国家重点研发计划(2022YFC3105203);国家自然科学基金(32072329);现代农业产业技术体系专项资金(CARS-49);中国水产科学研究院基本科研业务费(2023TDT6)共同资助。

摘　　要：麻痹性贝类毒素(paralytic shellfish toxins,PSTs)在我国乃至全球引发了严峻的生态和食品安全风险,加强该类毒素的安全监控以确保消费者安全成为全球共识。现有检测技术中,高效液相色谱-串联质谱技术因其高通量、高灵敏度的优势成为国际优先发展技术,但符合国际限量目标的参考物质获取成为发展中国家开展风险监测的核心难题。基于此,本研究通过比较6株产毒藻的单细胞产毒量、藻密度和毒素组分,筛选出的4株产毒藻所产毒素可以覆盖我国海域的主要PSTs,平均藻细胞密度达4.0×10^(6)~4.0×10^(7) cells/L。通过4株产毒藻单一暴露实验发现,贻贝中的PSTs含量能够满足国际限量目标,最高蓄积转化率可达83.1%,且藻与贝中毒素组成基本一致。此外,其中的链状裸甲藻可在贻贝体内代谢转化形成dcSTX,并在5 d达到29.6%的占比。根据上述规律,确定混合暴露实验时链状亚历山大藻(Alexandrium catenella)、链状裸甲藻(Gymnodinium catenatum)、塔玛亚历山大藻(A.tamarense)和微小亚历山大藻(A.minutum)投喂比例低密度组为1∶1∶1∶4,高密度组为1∶1.6∶2.4∶8,其中,高密度组的dcNEO、dcSTX、NEO等3种毒素的占比高于低密度组。4株产毒藻混合暴露获得含NEO、dcSTX、dcNEO、GTX1、GTX4、GTX2、GTX3、GTX5、dcGTX2、dcGTX3、C1和C2共12种PSTs,总毒性分别为(535.0±5.6)μg STXeq/kg和(2636.0±12.4)μg STXeq/kg的基体标准物质原料。本研究表明,产毒藻混合暴露可以获得稳定可控的12种PSTs毒素组分,为制备可用于产业监管及行业服务的基体标物提供技术支撑。Paralytic shellfish toxins(PSTs)pose significant ecological and food safety hazards in China and the world.Consequently,there is a global consensus to intensify safety monitoring of these toxins to safeguard consumers.In recent years,the occurrence of harmful algal blooms(known as“red tides”)has increased due to human activities and climate change.The contamination risk posed by PSTs is a pressing concern.This problem poses significant ecological and food safety risks in China and the world,leading to a global consensus on strengthening the safety monitoring of PSTs to ensure consumer safety.Several countries and regions,including the European Union,the United States,and Canada,have established strict regulatory limits(800μg STXeq/kg)for PSTs and implemented monitoring programs.Currently,the main methods for detecting and monitoring PSTs include enzyme-linked immunosorbent assay,mouse bioassay,high-performance liquid chromatography,and liquid chromatography-tandem mass spectrometry(LC-MS/MS).Among them,LC-MS/MS has gained international attention due to its high throughput and sensitivity,allowing detailed analysis of toxin profiles in contaminated shellfish.However,the complexity of aquatic food matrices often introduces interference during detection,affecting the accuracy and precision of the results.Using matrix reference materials as quality controls during the analysis can effectively mitigate the errors caused by matrix effects.In recent years,research has been conducted in China on the preparation techniques of matrix reference materials.However,existing techniques have limitations,such as unstable raw materials,limited toxin types,and lack of commercial production.There is still a lack of PST standard materials in China that possess traceability,certification,and physical specimens.To meet the quality control and validation needs for PST detection and analysis,and to enhance the regulation of shellfish quality and safety,the preparation of various PST matrix reference materials has become a top priori

关 键 词：贻贝 麻痹性贝类毒素 产毒藻 混合暴露 基体标准物质 

分 类 号：S917[农业科学—水产科学]