CAJ | 학술논문

背景:失忆性贝毒属于海洋神经毒素,是一类具有神经兴奋的氨基酸类物质,其主要成分为软骨藻酸.贝类通过滤食毒藻,将软骨藻酸富集在体内,人类因食用被软骨藻类污染的贝类而中毒.目的:建立液相色谱/四极杆-行时间质谱联用技术对贝类样品中的软骨藻酸实施有效监控,以期提高贝类的食用安全性.设计:前瞻性自身对照实验.单位:上海检验检疫局动植物与食品检验检疫技术中心理化实验室.材料:MUS-lB贻贝组织标样[(36±1)μg软骨藻酸/g,批号MUS-1B0159]和软骨藻酸标准溶液(100mg/L,批号软骨藻类CS-1C 352)购自National Research Council of Canada(Halifax,Canada).实际贻贝样品取自历年在东海爆发的赤潮海域收集得到的阳性贝类,空白样品购自市集.高效液相色谱仪(Waters 2695)配有紫外检测器(Waters 2487)和四极杆-飞行时间质谱仪(QUhima,Micromass).方法:样品用甲醇-水(体积比为1:1)提取,经SAX固相萃取柱净化后,用Zorbax Eclipse XDB-18柱(250×2.1 mm,3.5 μm)色谱分离,流动相为乙腈/水含0.05%甲酸,梯度淋洗,流速0.20 mL/min.四极杆-飞行时间质谱选择电喷雾正离子模式,以[M+H]+作为前体离子进行飞行时间质谱扫描测定.主要观察指标:①检测限.②液相色谱/四极杆-飞行时间质谱法的准确度与精密度.结果:样品加标的平均回收率为87.8%～94.6%,相对标准偏差为8.4%～11.9%,检测低限为0.1μg/g.结论:液相色谱/四极杆-飞行时间质谱法作为对贝类样品中失忆性贝毒的确证分析方法具有较高的灵敏度和选择性,其应用可为贝类食用的安全性监测提供参考指标.
배경:실억성패독속우해양신경독소,시일류구유신경흥강적안기산류물질,기주요성분위연골조산.패류통과려식독조,장연골조산부집재체내,인류인식용피연골조류오염적패류이중독.목적:건립액상색보/사겁간-행시간질보련용기술대패류양품중적연골조산실시유효감공,이기제고패류적식용안전성.설계:전첨성자신대조실험.단위:상해검험검역국동식물여식품검험검역기술중심이화실험실.재료:MUS-lB이패조직표양[(36±1)μg연골조산/g,비호MUS-1B0159]화연골조산표준용액(100mg/L,비호연골조류CS-1C 352)구자National Research Council of Canada(Halifax,Canada).실제이패양품취자력년재동해폭발적적조해역수집득도적양성패류,공백양품구자시집.고효액상색보의(Waters 2695)배유자외검측기(Waters 2487)화사겁간-비행시간질보의(QUhima,Micromass).방법:양품용갑순-수(체적비위1:1)제취,경SAX고상췌취주정화후,용Zorbax Eclipse XDB-18주(250×2.1 mm,3.5 μm)색보분리,류동상위을정/수함0.05%갑산,제도림세,류속0.20 mL/min.사겁간-비행시간질보선택전분무정리자모식,이[M+H]+작위전체리자진행비행시간질보소묘측정.주요관찰지표:①검측한.②액상색보/사겁간-비행시간질보법적준학도여정밀도.결과:양품가표적평균회수솔위87.8%～94.6%,상대표준편차위8.4%～11.9%,검측저한위0.1μg/g.결론:액상색보/사겁간-비행시간질보법작위대패류양품중실억성패독적학증분석방법구유교고적령민도화선택성,기응용가위패류식용적안전성감측제공삼고지표.
BACKGROUND: Amnesic shellfish poisoning(ASP) is caused by consumption of seafood contaminated by marine neurotoxins, mainly domoic acid (DA).OBJECTIVE: To ensure the safety of shellfish consumption by determining DA in shellfish by means of liquid chromatography(LC) combined with quadrupole time-of-flight mass spectrometry(Q-TOF MS).DESIGN: A self-controlled prospective study SETTING: The experiment was carried out in the Center for Animal, Plant and Food Inspection and Quarantine, Shanghai Exit-Entry Inspection and Quarantine Bureau.MATERIALS: MUS-1B, a nominal sample of homogenized mussel tissue (containing 36 ± 1μg DA per gram, Batch No. MUS-1B 0159) and the standard DA solution (100 mg/L, Batch No. DACS-1C 352) were purchased from National Research Council of Canada (Halifax, Canada). Positive musscl samples were collected from the waters during the occurrence of red tide in China East Sea, and blank mussel samples were obtained from local seafood market. LC system (Waters 2695) combined with Q-TOF MS (Qultima, Micromass) and ultravilet (UV) detection (Waters 2487 ) was employed for analysis of ASP toxins.METHODS: The shellfish tissue samples were extracted with methanol-water (1: 1, V/V) and cleansed with a SAX solid-phase extraction cartridge. Separation was carried out on a Zorbax Eclipse XDB-18 column (250×2.1 mm, 3.5μm) with gradient elution of CAN-H20 containing 0.05% formic acid at a flow rate of 0.20 mL per minute. In a positive electrospray ionization DA was ionized effectively, and collision-induced dissociation(CID) produced predominant characteristic ions[M + H] +allowing sensitive TOF MS detection.MAIN OUTCOME MEASURES: ① Limit of quantification(LOQ); ②Accuracy and precision of LC/Q-TOF MS method.RESULTS: The average recoveries of DA spiked to tissue homogenates through the complete cleanse procedure ranged from 87.8% to 94.6%, and the relative standard deviations(RSD) ranged from 8.4% to 11.9%. TheLOQ in the shellfish tissue was 0.1 μg/g.CONCLUSION: LC/Q-TOF MS suits well the purpose of detecting and identifying ASP toxins in shellfish with high sensitivity and specificity, and may provide reference values for monitoring the safety of consumption of shellfish.