河北省市售贝类中脂溶性毒素的调查研究

目的调查河北省市售贝类中脂溶性贝类毒素的污染状况。方法在2018年8月—2020年4月间,对河北省市售7大类贝类样品进行采集,共354份,采用超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)进行脂溶性贝类毒素的检测。结果 354份样品中,阳性样品17份,检出率为4.8%,检出含量较低,检出品种有毛蚶、贻贝、扇贝、牡蛎,毒素组分有虾夷扇贝毒素(yessotoxin,YTX)、同源虾夷扇贝毒素(homo-YTX)、大田软海绵酸(okadaicacid,OA)、鳍藻毒素(dinophysistoxins, DTXs)(DTX1、DTX2)、蛤毒素2(pectenotoxin-2, PTX2)。结论河北省市售贝类中脂溶性贝类毒素暴露风险很低。     

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。     

麻痹性贝类毒素监测指示贝种的筛选研究

为了解深圳地区贝类麻痹性贝类毒素(paralytic shellfish poison,PSP)染毒情况,对深圳地区主要贝类养殖海域养殖的10种贝类麻痹性贝类毒素染毒情况进行调查。筛选并确定PSP监测指示贝种,进而测定不同养殖海区PSP污染情况。采用AOAC发布的生物法测定麻痹性贝类毒素含量。华贵栉孔扇贝(Chalmys nobilis)和翡翠贻贝(Perna viridis)对PSP蓄积能力强,华贵栉孔扇贝的PSP含量较翡翠贻贝高。确定深圳贝类养殖海域PSP监测指示贝种为华贵栉孔扇贝和翡翠贻贝,全年对两种贝类的PSP监测结果表明PSP含量超过有关标准规定的安全限量的情况时有发生。本研究为实施有效的监测预警管理制度提供了科学依据。     

海南岛近岸贝类中麻痹性毒素污染情况调查研究

目的 为了调查清楚海南岛近岸海域麻痹性贝类毒素(paralytic shellfish poisons ,PSP)的污染状况。方法 对不同季节不同地域不同成分的毒素并进行调查研究。于2019年9月至2020年8月期间,在海南岛近岸海域设立6个采样点,涵盖海南东西南北不同区域,采集17种贝类共269份样本,利用液相色谱质谱法对样品进行麻痹性贝类毒素PSP的毒素测定和分析。结果 显示海南岛海域的贝类组织中PSP的含量检出率为15.2 %,结果均低于我国的警戒值（80 μgSTXeq/100g）。贝类中PSP的检出率随季节变化明显,以冬季最高,夏季次之,秋季最低;贝类中PSP的污染状况有明显的地域差别,东方、琼海、儋州、文昌海域的PSP污染状况较严重;贝类中检出的6种PSP成分,分别为漆沟藻毒素2（GTX2）、漆沟藻毒素3（GTX3）、漆沟藻毒素5（GTX5）、新石房蛤毒素（neoSTX）、脱氨甲酰基膝沟藻毒素（dcGTX2）和脱氨甲酰基石房哈毒素（dcSTX）。结论 海南岛近岸PSP的检出率不高,污染状况不是很严重,但还应长期监管,确保食用安全。     

超高效液相色谱-串联质谱法同时检测贝类中13种麻痹性贝类毒素

目的 建立超高效液相色谱-串联质谱法可以同时进行测定双壳贝类中13种麻痹性贝类毒素(paralytic shellfish poisoning toxins, PSTs)的方法。方法 用乙酸水溶液提取贝类中的PSTs,经乙酸乙酯除杂,然后用石墨化碳黑固相萃取柱净化,采用UPLC BEH Amide色谱柱 (100 mm×2.1 mm,1.7 μm)分离,电喷雾正、负离子多反应监测模式(MRM)同时检测,基质匹配曲线外标法定量。结果 13种PSTs的线性方程相关系数(r)均大于0.998,定量限为2 ~20 μg/kg,贻贝基质的平均回收率为92%~122%,牡蛎基质的平均回收率为105%~127%,扇贝基质的PSTs平均回收率为107%~144%,相对标准偏差均小于10%。将该方法应用于100份贝类样品中PSTs的检测,共有30份样品、5种PSTs检出,总毒性当量(STXeq)为10.0 ~159 μg/kg。结论 本研究方法灵敏度高,具有良好的重现性,可以准确定量定性13种PSTs ,适用于贝类中PSTs的日常监测。     

一起贝类膝沟藻毒素中毒调查分析

目的了解膝沟藻毒素中毒流行病学特征,探讨预防和救治麻痹性贝毒中毒的措施;方法对中毒者进行调查,采集样品、观察藻种并计数密度,检测菲律宾蛤仔毒力,测定麻痹性贝毒毒素成分;结果养殖菲律宾蛤仔的海水中亚历山大藻密度为5×105细胞/L,病人食剩的生、熟菲律宾蛤仔毒力分别为16 603 MU/100 g和4 035 MU/100 g,样品中均检出GTX1,GTX2,GTX3,GTX4。结论菲律宾蛤仔引起的中毒事件是一起贝类膝沟藻毒素中毒。     

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。     

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。     

基质分散固相萃取净化-亲水液相色谱-串联质谱法检测织纹螺与贝类中河鲀毒素

目的针对海产品常见中毒原因分析需求,建立基质分散固相萃取净化-亲水液相色谱-串联三重四极杆质谱(HILIC-MS/MS)快速定性定量检测织纹螺和贝类中河鲀毒素的新方法。方法 1.0 g样品经0.1%乙酸溶液沸水浴提取后,用50 mg亲水亲油平衡填料(HLB)和5 mg石墨化碳黑(GCB)吸附剂吸附净化,最后经乙腈蛋白沉淀后过0.22μm聚四氟乙烯(PTFE)滤膜,亲水液相色谱柱(150 mm×2.0 mm,3μm)分离,电喷雾离子化,选择反应监测(SRM)模式检测,基质匹配外标法定量。结果在2.0~40.0 ng/ml浓度范围内,河鲀毒素呈现良好的线性关系,相关系数r~2≥0.999;以3倍基线噪声所对应的浓度为检出限时,河鲀毒素的方法检出限可达10.0μg/kg;在25、100和200μg/kg的加标水平时,方法回收率为74.2%~87.9%,相对标准偏差为2.3%~9.1%。应用本方法对浙江沿海地区市售织纹螺和贝类样品进行检测,15份织纹螺中有14份检出河鲀毒素,检出率为93.3%,含量范围为0.04~15.75 mg/kg,60份贝类样品均未检出河鲀毒素。结论该检测方法准确、快速、易操作,能满足典型海产品中河鲀毒素的公共卫生应急检测或日常监测要求。     

高效液相色谱-串联质谱法测定双壳类水产品麻痹性贝类毒素

目的采用Supelco ENVI-Carb柱净化双壳类水产品中的麻痹性贝类毒素(PSP),建立高效液相色谱-串联质谱(HPLC-MS/MS)法检测双壳类水产品中的PSP,为水产品中的PSP检测提供方法依据。方法选用色谱柱TSK-GEL Amide-80(2.0 mm×250 mm,5μm),以2 mmol/L甲酸铵-50 mmol/L甲酸水和95%乙腈水(含2 mmol/L甲酸铵-50 mmol/L甲酸)为流动相,采用梯度洗脱进行分离。样品用1%乙酸溶液进行提取,上清液加入氨水后(pH=4.0)经Supelco ENVI-Carb柱净化,将洗脱液抽干收集,上机测定。多重反应监测(MRM)模式检测。结果PSP的线性范围为8.1~705.0μg/kg,检出限为10~35μg/kg,回收率在47.0%~91.3%之间。结论本方法提取效果好、基质效应小,适用于双壳类水产品中麻痹性贝类毒素的痕量检测。     

秦皇岛海域食用贝类重金属污染情况分析

为了解秦皇岛海域食用贝类中重金属的污染水平及其存在的健康风险。采用电感耦合等离子体发射质谱法(ICP-MS)和原子荧光光谱法测定了该地区15种常见贝类肌肉和内脏中As(无机砷和总砷)、Pb(铅)、Cr(铬)、Cd(镉)、Hg(甲基汞)的含量,并采用污染指数法、系统聚类法评价了各重金属的污染特征和分类。结果表明:海虹(贻贝,Mytilus edulis)、栉孔贝(Chlamys farreri)、生蚝(Ostrea gigas thunberg)和毛蚶子(Scapharca subcrenata)等的重金属综合污染(MPI)较高。根据NY 5073-2006 无公害食品水产品中有毒有害物质限量标准进行重金属评价。秦皇岛海域食用贝类六种重金属含量最高为Cd,均值为1.129 mg/kg,超出NY 5073-2006 无公害食品水产品中有毒有害物质限量标准,污染较严重,总As和无机As均值分别为0.161、0.021 mg/kg。Cr和Hg均值分别为0.107和0.013 mg/kg,均未受污染,Pb均值为0.188 mg/kg,污染较轻。但部分贝类Cd污染比较严重。其中内脏污染比肌肉部分严重,尤其是大小海螺(Busycon canaliculatu)。因此,需进一步研究秦皇岛海域可食用贝类中Pb和Cd的污染来源并进行有效防治。     

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.     

First paralytic shellfish poison (PSP) infestation of bivalves due to toxic dinoflagellate Alexandrium tamiyavanichii, in the southeast coasts of the Seto Inland Sea, Japan

The mussel Mytilus edulis and the cultured ark shell Anadara broughtonii in the southeast coasts of the Seto Inland Sea were contaminated with paralytic shellfish poison (PSP) following the appearance of the dinoflagellate Alexandrium tamiyavanichii in early December 1999. A. tamiyavanichii plankton collected around the Straits of Naruto on December 3, 1999 showed PSP toxicity, of which 83 mol% was accounted for by GTX2, GTX3 and GTX4. Its specific toxicity was 112.5 fmol/cell, and one MU was equivalent to 7,200 cells. Toxicity values at the beginning of toxification were 4.7 MU/g for the ark shell and 7.3 MU/g for the mussel. In the former, the value remained at almost 4 MU/g, resulting in prohibition of marketing for about two months. In the latter, it sharply decreased to less than 4 MU/g. These bivalves collected during the toxification period were dissected into five tissues, mantle, adductor muscle, hepatopancreas, gills and "others", and submitted to high-performance liquid chromatography (HPLC). The cultured ark shell accumulated GTX2, GTX3 and STX as major components and GTX1, GTX4, GTX5, neoSTX, dcSTX and PX1-3 (C1-C3) as minor ones. The amount of GTX3 decreased with time, while STX tended to increase. At the early stage of PSP toxification, toxins were accumulated in the gills and "others", most of which were quickly detoxified. On the other hand, PSP of the toxified mussel consisted of GTX4 as a main component, and GTX1, GTX2, GTX3, GTX5, STX and PX1-2 (C1-C2) as minor ones. Its toxin composition pattern was similar to that of the ingested causative plankton. Its total toxin decreased soon after disappearance of the dinoflagellate. During the decrease of toxicity, PSP tended to be retained in the hepatopancreas, resulting in accumulation of 50 mol% of total toxin.     

Analysis of paralytic shellfish toxins and their metabolites in shellfish from the North Yellow Sea of China

Samples of toxic scallop (Patinopecten yessoensis) and clam (Saxidomus purpuratus) collected on the northern coast of China from 2008 to 2009 were analysed. High-performance liquid chromatography with post-column oxidation and fluorescence detection was used to determine the profile of the main paralytic shellfish poisoning (PSP) toxins in these samples and their total toxicity. Hydrophilic interaction liquid ion chromatography with mass spectrometric detection confirmed the toxin profile and detected several metabolites in the shellfish. Results show that C1/2 toxins were the most dominant toxins in the scallop and clam samples. However, GTX1/4 and GTX2/3 were also present. M1 was the predominant metabolite in all the samples, but M3 and M5 were also identified, along with three previously unreported presumed metabolites, M6, M8 and M10. The results indicate that the biotransformation of toxins was species specific. It was concluded that the reductive enzyme in clams is more active than in scallops and that an enzyme in scallops is more apt to catalyse hydrolysis of both the sulfonate moiety at the N-sulfocabamoyl of C toxins and the 11-hydroxysulfate of C and GTX toxins to produce metabolites. This is the first report of new metabolites of PSP toxins in scallops and clams collected in China.     

贝类毒素检测中两种筛选方法的比较研究

目的 采用传统小鼠生物法(MBA)和酶联免疫吸附法(ELISA)对贝类样品中四类毒素进行检测, 为不同要求下建立或选择准确的贝类毒素检测快速筛选方法提供参考。方法 分别采用MBA和ELISA检测腹泻性贝毒(DSP)和麻痹性贝毒 (PSP), 并采用ELISA检测记忆缺损性贝毒(ASP) 和神经性贝毒(NSP)。结果 对2009~2011年8种67份贝类样品进行检测, 结果表明: 两种测试方法在实际应用中对DSP、PSP检测结果不存在差异, 检测结果有很好的吻合性。使用ELISA法对自制ASP、NSP模拟阳性样品进行检测, 均测得ASP、NSP, 检测结果满意。结论 两种筛选方法在贝类毒素检测中均有其应用空间。实验室可根据不同情况选择合适的检测方法。     

天津市市售贝类腹泻性贝类毒素的调查研究

目的 对2011年2月-2012年1月天津市售双壳经济贝类腹泻性贝毒(DSP)污染状况进行为期一年的抽样,调查其食用安全性.方法 分别采用小鼠生物法和液相色谱-串联质谱法(LC/MS-MS)对可食部分进行检测,测定毒素含量及分析毒素组成.结果 在所抽查的10种103个贝类样品中,只有产自河北唐山的毛蚶中5月和8月采集的样品中有2个呈阳性且均超标.对阳性样品进行LC/MS-MS检测,检出DTX-1和YTX毒素.结论 腹泻性贝毒在天津市场范围内检出率及含量整体水平不高,但应提示有关部门应在春季预防,在夏季加强对食用贝类的监测和宣传,以保证市民健康安全.     

Matrix effect on paralytic shellfish toxins quantification and toxicity estimation in mussels exposed to Gymnodinium catenatum

Paralytic shellfish toxins were quantified in whole tissues of the mussel Mytilus galloprovincialis exposed to blooms of the dinoflagellate Gymnodinium catenatum in Portuguese coastal waters. A validated liquid chromatography method with fluorescence detection, involving pre-chromatographic oxidation was used to quantify carbamoyl, N-sulfocarbamoyl and decarbamoyl toxins. In order to test for any matrix effect in the quantification of those toxins, concentrations obtained from solvent and matrix matched calibration curves were compared. A suppression of the fluorescence signal was observed in mussel extract or fraction in comparison to solvent for the compounds dcGTX2 + 3, GTX2 + 3 and GTX1 + 4, while an enhancement was found for C1 + 2, dcSTX, STX, B1, dcNEO and NEO. These results showed that a matrix effect varies among compounds. The difference of concentrations between solvent and matrix matched calibration curves for C1 + 2 (median = 421 ng g?1) exceeded largely the values for the other quantified compounds (0.09-58 ng g?1). Those differences were converted into toxicity differences, using Oshima toxicity equivalence factors. The compounds C1 + 2 and dcNEO were the major contributors to the differences of total toxicity in the mussel samples. The differences of total toxicity were calculated in ten mussel samples collected during a 10-week blooming period in Portuguese coastal lagoon. Values varied between 53 and 218 μg STX equivalents kg?1. The positive differences mean that the estimated toxicity using solvent calibration curves exceed the values taking into account the matrix. For the toxicity interval 200-800 μg STX equivalents kg?1 an increase was found between 44 and 28%.