海产品中麻痹性贝类毒素快速检测技术研究进展

麻痹性贝类毒素是我国海洋赤潮中最常见的贝类毒素之一,分布最广,危害最大,事故发生率也最高,对人类健康构成了严重威胁,加强对该类毒素的检测监控成为保障海产品安全的重要措施。传统的检测方法主要有小鼠生物检测法、液相色谱法、液相色谱-串联质谱法和酶联免疫法,这些方法均有各自的优势,但在实际应用中还缺少用于现场检测的快速筛查技术。因此,开发快速、灵敏、准确、低成本的麻痹性贝类毒素检测技术具有重要的应用价值。本文主要介绍了麻痹性贝类毒素目前开发出来的快速检测方法,主要包括免疫层析技术和生物传感器技术,对各方法的特点迚行分析。最后对未来麻痹性贝类毒素快速检测技术在实际应用中面临的主要问题迚行了评述,幵对发展趋势迚行了展望。     

麻痹性贝类毒素常规检测分析方法比较与 研究进展

麻痹性贝类毒素作为贝类产品中一种毒性最强、分布广泛的毒素,不仅严重威胁人们的身体健康,而且会造成相当大的经济损失。因此其监测检测方法的研究与改进一直是人们的研究热点。本文分析评述了麻痹性贝类毒素的三种常规检测分析方法的优缺点以及最新研究进展,并探讨了小白鼠生物法、免疫测定法和色谱联用技术作为主要的检测方法由于原理不同,结合不同的研究需求其应用的领域。其中,小白鼠生物测定法虽然概括毒性有效,但是其灵敏度低、误差大、并且需要大量活体动物而逐渐被色谱技术和免疫测定法所取代,此外,神经细胞分析法、毛细管电泳技术和表面等离子体共振传感器技术等方法也逐渐得到应用。不管怎样,这些方法由于需要专业人员、成本高等问题仍需进一步完善。     

认识麻痹性贝类毒素

正织纹螺因含有麻痹性贝类毒素而被国家禁止食用。麻痹性贝类毒素是如何产生的?如何避免?既然称其为"贝类毒素",我们平时所吃的美味贝类都安全吗?麻痹性贝类毒素是一种生物毒素麻痹性贝类毒素并非来自贝类生物体本身,而是贝类摄食有毒藻类,并经其体内蓄积、放大、转化等过程形成的具有神经肌肉麻痹作用的赤潮生物毒素。人体若误食含有此类毒素的贝类     

贝类毒素及检测方法的研究进展

针对腹泻性贝毒（DSP）、麻痹性贝毒（PSP）、神经性贝毒（NSP）和记忆缺损性贝毒（ASP）等4种主要贝类毒素，描述了其主要特征、毒性大小及毒理效应，对已有的生物分析检测方法和仪器分析技术作介绍。     

麻痹性贝类毒素常规检测分析方法比较与研究进展

麻痹性贝类毒素作为贝类产品中一种毒性最强、分布广泛的毒素,不仅严重威胁人们的身体健康,而且会造成相当大的经济损失。因此其监测检测方法的研究与改进一直是人们的研究热点。本文分析评述了麻痹性贝类毒素的三种常规检测分析方法的优缺点以及最新研究进展,并探讨了小白鼠生物法、免疫测定法和色谱联用技术作为主要的检测方法由于原理不同,结合不同的研究需求其应用的领域。其中,小白鼠生物测定法虽然概括毒性有效,但是其灵敏度低、误差大、并且需要大量活体动物而逐渐被色谱技术和免疫测定法所取代,此外,神经细胞分析法、毛细管电泳技术和表面等离子体共振传感器技术等方法也逐渐得到应用。不管怎样,这些方法由于需要专业人员、成本高等问题仍需进一步完善。     

连云港海州湾麻痹性贝类毒素中毒分析

为分析连云港地区麻痹性贝类毒素中毒特征,对连云港海州湾织纹螺进行形态鉴定,收集近年麻痹性贝类中毒资料,应用小白鼠生物测定法检测螺肉中麻痹性贝类毒素。结果连云港海州湾存有4种织纹螺,其中以半褶织纹螺含麻痹性贝类毒素量高(1.6×103μg100g螺肉),并导致连云港地区10起、银川1起麻痹性贝类中毒;1992年还发生一起由泥螺引起的麻痹性贝类中毒。中毒者均表现为下行性神经麻痹症状,重者致死。鉴于海洋污染的严重性,为保障海洋贝类的食用安全,应对贝类进行毒素等安全指标的监测。     

硅胶性能表征及其对麻痹性贝类毒素吸附研究

研究硅胶对麻痹性贝类毒素吸附性能情况,探究其作为毒素检测前处理材料的可能性。利用扫描电子显微镜、红外光谱分别表征硅胶形貌尺寸、官能团与化学键,使用差示扫描量热仪、热重分析仪分析硅胶的热稳定性,采用液相色谱-串联质谱研究硅胶对麻痹性贝类毒素标准溶液吸附情况。结果表明,所选用硅胶直径介于30～60μm,尺寸相对均一,富含硅羟基团,热稳定性好,在弱酸或中性条件下有效吸附麻痹性贝类毒素,并能在0.1%酸性条件下解吸附。鉴于硅胶自身特性及对麻痹性贝类毒素良好的吸附能力,有望用于动物性水产食品中麻痹性贝类毒素检测净化前处理,其吸附机理是表面的硅羟基与麻痹性贝类毒素分子中氨基、亚氨基之间形成氢键的作用。     

钦州湾7种经济贝类中麻痹性贝类毒素分析与安全评价

采用小鼠生物检测法和液相色谱-荧光检测法,分别对采自钦州湾的7种典型经济贝类中麻痹性贝类毒素(paralytic shellfish toxins,PSTs)组成成分与含量进行分析,同时参考我国渔政渔港监督管理局制订的贝类安全食用标准(400 MU/100 g或80μg/100 g STX_(eq))评价其食用安全性。结果如下,小鼠生物法分析表明小鼠在观察15 min内均不死亡,说明7种贝类中PSTs含量均小于400 MU/100 g或者不含有毒素;进一步对贝类样品进行液相色谱-荧光检测分析,表明7种贝类中可检测到麻痹性贝类毒素,毒素组成成分以高毒性的氨基甲酸酯类毒素为主如膝沟藻毒素4(gonyautoxin,GTX4)、膝沟藻毒素1(gonyautoxin,GTX1)、新石房哈毒素(neosaxitoxin,NEO)和石房蛤毒素(saxitxin,STX),其中异毛蚶中有最大毒素含量/毒性值,分别为0.27 nmol/g或13.1μg/100 g STX_(eq),低于安全标准80μg/100 g STX_(eq)。研究中有6种贝类可检测到毒素成分,检出率为86%。这说明,在钦州湾所采集的7种贝类中,尽管毒素含量低于食用安全标准规定的阈值,但多种贝类中仍可检测到麻痹性贝类毒素成分,因此其存在的安全性问题不容忽视,应加强该地区经济型贝类中麻痹性贝类毒素的监测,以防中毒事件的发生。     

上海市售贝类产品中麻痹性贝类毒素污染状况调查及其评价

目的 调查上海市售贝类产品中麻痹性贝类毒素污染状况。方法 2010年8月~2011年7月间, 在上海水产品批发市场进行5种贝类样品采集, 每月抽取样品24份, 全年共288份。采用生物法(SC/T 3023-2004)对其进行了麻痹性贝类毒素的检测, 其中虾夷扇贝的肠腺和肌肉(扇贝柱)进行分开测定。结果 缢蛏、菲律宾蛤仔、牡蛎、文蛤、虾夷扇贝肠腺和肌肉中麻痹性贝类毒素的含量范围分别为ND~121.5 MU/100 g、ND~113.4 MU/100 g、ND~177.7 MU/100 g、ND~124.6 MU/100 g、261.7~3363.5 MU/100 g和ND。全年麻痹性贝类毒素的平均含量分别在98.5±10.5 MU/100 g、78.6±9.3 MU/100 g、50.4±10.1 MU/100 g、40.6±14.8 MU/100 g、1242.2±974.3 MU/100 g和0。按照目前我国贝类产品主要出口国家和国际组织对麻痹性贝类毒素的限量要求进行评价, 仅仅是虾夷扇贝肠腺中麻痹性贝类毒素超标, 超标率为98%, 因此在食用扇贝时应去除其肠腺; 而其余贝类产品中麻痹性贝类毒素均在限量规定范围内。结论 上海市售贝类产品对食用的安全性不产生负面影响。     

舟山海域麻痹性贝类毒素污染情况及其2种检测方法比较

目的检测舟山东极与嵊泗枸杞2个海域养殖贻贝中的麻痹性贝类毒素(paralytic shellfish poison,PSP),比较小鼠生物测定法与酶联免疫分析法(ELISA)的测定结果。方法采用小鼠生物测定法与酶联免疫吸附法检测贝类中的麻痹性贝类毒素,并将2者的检测结果进行比较分析。结果 2种检测方法检测的麻痹性贝类毒素含量结果基本一致。5月份东极岛海域的厚壳贻贝中检出PSP((500±3.2)MU/100 g),超标率为5%;嵊泗枸杞海域贝类PSP含量较低,未超出安全食用标准。2个海域的紫贻贝PSP含量均未超出安全食用标准。结论小鼠生物法与ELISA方法的评价结果基本一致,其检测出的PSP结果可以为摄入PSP风险评估提供数据支撑。由于ELISA方法的检测成本较高,因此可采用小鼠生物法进行麻痹性贝类毒素风险监测。     

Keeping shellfish safe to eat: a brief review of shellfish toxins,and methods for their detection

Poisoning can result from the ingestion of shellfish contaminated with phycotoxins. Various types of poisoning may occur, each of which is caused by a toxin (or group of toxins) from a particular alga. Classically, the mouse bioassay has been used to detect shellfish toxins, but there is pressure, both ethical and regulatory, to move away from this. A number of techniques have been developed to replace the bioassay, including immunoassay, chromatography, pharmacological assay and tissue-culture tests. All have advantages and limitations. These methods and their potential are reviewed.     

Rapid Detection of Chemical Hazards (Toxins,Dioxins, and PCBs) in Seafood

Among the various hazards occurring in fish and seafood chemical hazards and in particular toxins (ciguatera, scombroid fish poisoning, paralytic shellfish poisoning, neurotoxic (brevetoxic) shellfish poisoning, puffer fish poisoning, diarrhetic shellfish poisoning) have an important place in food poisoning cases. On the other hand, some of the chemical hazards are often due to the pollution of the environment (heavy metals, dioxins, polychlorinated biphenyls, and halogenated aromatic hydrocarbons) and their detection is neither rapid nor facile. As a result there was a great need for developing new rapid and effective methods toward the chemical hazards determination mainly because of their high toxicity. The aim of this review is to provide the information about the new up-to-date detection techniques (Immunological, Chemical and Biochemical, and Molecular assays) in conjunction with detection limits. The latter is made possible by means of inclusion of seven comprehensive and, in most case cases, very extended tables. A reference is also made on the risk characterization of toxins as regards their importance to food contamination or poisoning.     

贝类毒素大田软海绵酸的免疫分析检测技术研究进展

大田软海绵酸(okadaicacid,OA)是一种广泛存在于贝类等生物中的海洋生物毒素,可引起人或动物的急性中毒,对食品安全和海产养殖具有严重危害。因此建立快速、可靠、灵敏的OA检测技术具有重要意义。免疫分析检测技术基于抗原抗体的结合,特异性强、灵敏度高、应用范围广,是当前检测贝类毒素OA的主要手段。本文综述了近年来针对贝类毒素OA的免疫分析检测技术,其中包括酶联免疫吸附检测、免疫层析检测、时间分辨荧光免疫检测和基于免疫传感器的检测技术等。本文着重阐述了不同免疫分析技术的原理及其在OA检测中的实际应用,同时探讨了免疫分析技术在贝类毒素OA检测方面的挑战和发展趋势,以期为开发性能更加优异的OA免疫检测技术提供研究思路。     

Contamination of shellfish from Shanghai seafood markets with paralytic shellfish poisoning and diarrhetic shellfish poisoning toxins determined by mouse bioassay and HPLC

This paper reports the results of investigations of shellfish toxin contamination of products obtained from Shanghai seafood markets. From May to October 2003, 66 samples were collected from several major seafood markets. Paralytic shellfish poisoning (PSP) and diarrhetic shellfish poisoning (DSP) toxins in shellfish samples were monitored primarily by a mouse bioassay, then analysed by HPLC for the chemical contents of the toxins. According to the mouse bioassay, eight samples were detected to be contaminated by PSP toxins and seven samples were contaminated by DSP toxins. Subsequent HPLC analysis indicated that the concentrations of the PSP toxins ranged from 0.2 to 1.9 µg/100 g tissues and the main components were gonyautoxins 2/3 (GTX2/3). As for DSP, okadaic acid was detected in three samples, and its concentration ranged from 3.2 to 17.5 µg/100 g tissues. Beside okadaic acid, its analogues, dinophysistoxins (DTX1), were found in one sample. According to the results, gastropod (Neverita didyma) and scallop (Argopecten irradians) were more likely contaminated with PSP and DSP toxins, and most of the contaminated samples were collected from Tongchuan and Fuxi markets. In addition, the contaminated samples were always found in May, June and July. Therefore, consumers should be cautious about eating the potential toxic shellfish during this specific period.     

腹泻性贝类毒素DSP的磷酸酶抑制检测法研究

对腹泻性贝类毒素DSP的快速筛选方法——磷酸酶抑制法进行了研究,分析了该方法的检测原理,并提出了该方法的筛选检出限,对标准溶液、加标回收样品及市场采样样品进行了检测,并就该方法的重复性进行验证。检测结果显示,磷酸酶抑制法检测准确率高,重复性较好,提出的筛选检出限（200μg/kg）远低于我国相关标准的规定（600μg/kg）,是一种对于腹泻性贝类毒素DSP较为理想的筛选方法。     

Comparison of HPLC and Mouse Bioassay Methods for Determining PSP Toxins in Shellfish

Paralytic shellfish poisoning (PSP) toxins in shellfish were analyzed and quantified using both a high pressure liquid chromatographic (HPLC) technique and the standard AOAC mouse bioassay. Good correlation between the two assays was obtained at or below the limit of 80 μg toxin/100g, while at higher levels of toxicity, the HPLC technique tended to underestimate total toxicity slightly. Advantages and disadvantages of both techniques are discussed.     

海洋生物毒素的分类、毒害作用机制及检测技术研究进展

近年来,海洋生物毒素引起的食源性中毒事件数量逐年上升,对人类和社会造成了较大危害。海洋生物毒素主要由藻类、浮游植物或微生物产生,可在贝类、鱼类等海洋生物体内长时间存在,具有耐热性,故加热、微波等常规加工方式无法使其失活;当其通过食物链进入人体后,极易对肝脏、细胞、心血管系统及神经系统等产生毒副作用,从而引起人体中毒。本文对海洋生物毒素的分类、毒害作用机制以及相应的检测技术进行综述分析,以期为防控海洋生物毒素对人体健康产生危害提供一定的参考价值。     

Hydrolysis of hydroxybenzoate saxitoxin analogues originating from Gymnodinium catenatum

The paralytic shellfish poisoning (PSP) toxin producer Gymnodinium catenatum produces several hydrophobic analogues of saxitoxin (STX). These are poorly studied due to their recent discovery and lack of standards. It was previously observed these hydrophobic analogues could be partially hydrolysed, loosing its benzoate moiety during alkaline oxidation to obtain fluorescent products measurable by HPLC analysis. The hydrolysis reaction was further explored to study two practical aspects. One was the indirect measurement of these compounds through its hydrolysis products: the decarbamoyl analogues of STX. The second one was to simplify standard production of decarbamoyl analogues, which are commonly found in contaminated shellfish products.