青岛市售贝类海洋生物毒素监测及其健康风险评估

发布时间：2018-03-16 03:16

本文选题：液相色谱串联三重四级杆(LC-MS/MS)　切入点：亲水性贝类毒素　出处：《青岛大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:1.本研究课题建立14种亲水性贝类毒素和11种亲脂性贝类毒素的测定方法。2.测定当前青岛市市售牡蛎、扇贝等贝类海产品中亲水性贝类毒素和亲水性贝类毒素的种类和含量,分析其时间变化规律,并进行风险评估。方法:1.14种亲水性贝类毒素和11种亲脂性贝类毒素测定方法的建立1%乙酸加热提取样品,再由石墨化炭黑固相萃取柱净化后,液相色谱-串联质谱测定麻痹性贝类毒素,外标法定量。样品组织经100%甲醇提取,C18固相萃取柱净化后,直接进样用于游离态的亲脂性贝类毒素以及软骨藻酸测定,基质加标校正,外标法定量。2.贝类海产品中贝类毒素的测定采集贝类样品包括牡蛎、杂色蛤、缢蛏、虾夷扇贝、栉孔扇贝、贻贝、毛蚶、海螺、钝拟蟹守螺、香螺和香波螺等样品,12批次共计400个,样品分成两部分,一部分利用1%乙酸加热提取,石墨化炭黑固相萃取柱净化;另一部分则利用甲醇提取,C18固相萃取柱净化提取,3.膳食消费量调查按24小时膳食回顾法问卷调查进行膳食习惯调查,对青岛市某区的1104名居民进行调查,连续三天调查居民贝类食品消费量,记账法记录消耗量数据。4.结果分析方法计算采集样品中贝类毒素种类和含量,分析不同时间样品中贝类毒素的含量分布情况。采用日暴露法,计算膳食摄入贝类毒素的日膳食暴露量并进行风险评估。结果:1所建立的方法25种贝类毒素检出限为4ug/kg~711ug/kg,精密度为0.8%~7.6%,平均回收率为85.2%~105.5%。2检测样品中贝类毒素种类和含量,其时间变化规律以及风险评估2.1检测样品中贝类毒素种类和含量,其时间变化规律本研究检测12批次共计400份样品,其中有73份有检出,以亲脂性贝类毒素为主,其检出率为18.50%。其中,主要检出成分有软海绵酸(okadaic acid,OA)、蛤毒素(pectenotoxin-2,PTX-2)和虾夷扇贝毒素YTXs(Yesssotoxin),其他种类的贝类毒素尚未检出。检出样品中以扇贝和贻贝为主,其余种类样品未检出。贝类样品的阳性检出率随时间变化而呈现一定差异,5-8月份贝类样品中腹泻性贝类毒素检出率最高。OA在8月份检出率最高,达29.20%,YTX在8月份检出率最高,达26.80%,PTX-2在7月份检出率最高,达35.20%,其中PTX-2最高含量为7.189μg/kg,尚未检测出其他贝类毒素。2.2风险评估食用贝类体中的亲脂性贝类毒素OA、PTX-2、YTX日暴露(摄人)量分别为0.48、0.88、0.65,按照膳食调查食用量水平下,尤其是夏秋季有可能会出现OA急性中毒事件,长期大剂量暴露其中有可能会威胁健康。食用含有YTX的贝类暴露量均在安全范围内,暂时没有其他危险影响。结论:1.所建立方法稳定,回收率符合实验要求,操作较为简单,灵敏度和精密度满足检测确证。2.不同种贝类中毒素种类和含量不同,不同季节中贝类毒素含量不同。3.大多数贝类中存在贝类毒素污染的问题,根据检测的目前市面贝类样品中毒素的含量小于欧盟限量值,对人们生活暂时不构成危害风险,但后续长期受到污染对人类健康是否存在影响有待进一步研究。
[Abstract]:Objective: a.2. method for determination of 1. this paper establishes 14 kinds of hydrophilic and lipophilic shellfish toxins 11 kinds of shellfish toxin determination currently sold in Qingdao City oysters, species and content such as hydrophilic Scallop in Shell shellfish shellfish toxins and hydrophilic shellfish toxins, analyze the time variation and risk assessment. Methods: extraction sample build 1% acetic acid heating method for the determination of 1.14 kinds of hydrophilic and lipophilic shellfish toxins 11 kinds of shellfish toxins, then by graphite carbon black solid phase extraction column purification, determination of paralytic shellfish toxin liquid chromatography tandem mass spectrometry, and quantified by external standard method. The sample tissue was extracted with 100% methanol, purified by C18 solid phase extraction column, directly the sample for free and lipophilic shellfish toxin domoic acid determination, matrix calibration, external standard method for quantitative.2. of shellfish toxins in shellfish seafood were collected in shellfish samples including oyster, Clam, Sinonovacula constricta, Yesso Scallop in Shell Scallop in Shell, Chlamys, mussel, clam, conch, blunt quasi cerithium, conch and shampoo snail samples, a total of 12 batches of 400 samples, divided into two parts, one part with 1% acetic acid extraction, graphitized carbon black solid phase extraction column purification; the other part is using methanol extraction, C18 solid phase extraction column purification extraction, 3. dietary consumption survey in 24 hour dietary recall questionnaire for dietary habits survey, conducted a survey of 1104 residents in a district of Qingdao City, three consecutive days of investigation of resident shellfish consumption accounts record data analysis of the results of.4. method to calculate the sample in shellfish species and content consumption, content distribution analysis of shellfish toxin in different time samples. By day exposure, dietary intake calculation of shellfish toxin exposure and risk assessment. Results: 1 of 25 Kind of shellfish toxin detection limit was 4ug/kg~711ug/kg, the precision is 0.8%~7.6%, the average recovery rate of 85.2%~105.5%.2 in the samples of shellfish toxin type and content, the time variation of risk assessment and 2.1 samples of shellfish toxin types and content of this study is to detect the time variation of the total of 12 batches of 400 samples, of which 73 samples are detected the lipophilic shellfish toxins, and the detection rate of 18.50%. among them, the main components of detection of okadaic acid (okadaic acid, OA), saxitoxin (pectenotoxin-2, PTX-2) and mizuhopecten Scallop in Shell toxin YTXs (Yesssotoxin), other types of shellfish toxins have not been detected. The detection of samples to Scallop in Shell and mussels, other species the samples were not detected in shellfish samples. The positive rate of change with time and showed some differences in the detection of diarrhetic shellfish poison in the highest rate of.OA in August was 5-8 in shellfish samples The highest rate reached 29.20%, the highest detection rate of YTX in August, PTX-2 in July reached 26.80%, the highest positive rate was 35.20%, the highest content of PTX-2 was 7.189 g/kg, has not yet been detected in other shellfish toxin.2.2 risk assessment of edible shellfish in lipophilic shellfish toxins OA, PTX-2, YTX (intake) amount of exposure are 0.48,0.88,0.65, the level of consumption in accordance with the dietary survey, especially there may be OA in acute poisoning in summer and autumn, the long-term high dose exposure which may be a threat to health. Food containing YTX shellfish exposure in the safe range, no effect. Other dangerous conclusion: 1. the method is stable, the recovery rate in line with the experimental requirements, the operation is simple, sensitivity and precision meet the test confirmed.2. of different types and different hormone content of shellfish poisoning, shellfish toxin content in different seasons have different.3. most in shellfish The pollution problem of shellfish poison is based on the detection of toxins in the market shellfish, which is less than the European limit. It does not pose a risk to people's lives. But whether the long-term pollution is affecting human health needs further study.

【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R155.5

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