固相萃取-高效液相色谱-串联质谱法测定贝类产品中麻痹性贝类毒素  

Determination of paralytic shellfish poisoning toxins in shellfish by high performance liquid chromatography-tandem mass spectrometry with solid phase extraction

作　　者：陈剑刚[1] 朱炳辉[2] 梁素丹 张瑰[1] 吴西梅[2] 

机构地区：[1]珠海市疾病预防控制中心,广东珠海519060 [2]广东省疾病预防控制中心,广东广州511430

出　　处：《中国卫生检验杂志》2016年第1期4-8,共5页Chinese Journal of Health Laboratory Technology

基　　金：珠海市科技计划项目(2013D0401990022)

摘　　要：目的建立了贝类产品中石房蛤毒素(STX)、新石房蛤毒素(neo STX)、膝沟澡毒素(GTX1&4)、脱氨甲酰基类毒素(包括dc STX、dcneo STX和dc GTX2&3)8种麻痹性贝类毒素的固相萃取-高效液相色谱-串联质谱测定方法。方法匀浆贝组织以0.1 mol/L乙酸提取,HLB固相萃取小柱净化,10 000 MW超滤离心管离心纯化。采用TSK gel Amide-80柱分离,以乙腈-2 mmol/L乙酸铵溶液(均含0.1%甲酸)为流动相进行梯度洗脱,以正离子多反应监测模式检测,基质标准校正外标法定量。结果 8种麻痹性贝类毒素的定量限为10μg/kg^50μg/kg;在各自相应浓度范围内线性关系良好,相关系数>0.990;2种添加水平的平均加标回收率为75.5%~103.0%;相对标准偏差为4.3%~13.2%。应用建立的方法对多份贝类样品进行分析,均未检出目标组分。结论方法选择性高、灵敏、准确,适用于贝类产品中麻痹性贝类毒素的确证及定量分析。Objective A solid phase extraction coupled to high performance liquid chromatography -tandem mass spectrometry （HPLC -MS/MS） method has been developed for the determination of 8 paralytic shellfish poisoning（PSP） toxins, including Saxitoxin （ STX）, Neo saxitoxin （ neoSTX）, knee ditch bath （ GTX1 ＆4）, toxin removal of ammonia formyl toxoid （ including dc- STX, dcneoSTX and dc GTX2＆3）. Methods Homo genised tissues were extracted with 0.1 mol/L HAc, and resulting extracts were cleaned - up on oasis HLB solid phase extraction column and filtered through a 10 000 MW uhra - filter centrifuge tube. The target components were separated and gradient elnted on a TSK gel Amide - 80 column using a mobile phase of MeCN - 2 mmol/L ammonium acetate, both phase containing 0. 1% formic acid, and simultaneously quantified by operating in positive ions under multiple reactions monitoring mode. The matrix - matched external standard calibration curves passing through extraction and clean -up procedures were carried out for the quantitative analysis. Results The limits of quantification ranged from 10 μg/kg to 50 μg/kg for 8 PSP toxins. The good linearity was obtained in the respective concentration ranges, with the correlation coefficients 〉 0.990. The average recoveries for toxins at two concentration levels were among 75.5% - 103.0%, and their relative standard deviation varied among 4.3% - 13.2%. The samples were analyzed in the application of the method, and the target components were not detected. Conclusion The method was selective, sensitive and accurate, and was suitable for confirmation and quantitative analysis of PSP toxins in shellfish.

关 键 词：高效液相色谱-串联质谱法 麻痹性贝类毒素 固相萃取 贝类产品 

分 类 号：O657.63[理学—分析化学]