使用免疫亲和柱净化、柱后衍生化和液相色谱/荧光检测法测定橄榄油、花生油和芝麻油中的黄曲霉毒素B1、B2、G1和G2：协同研究

Determination of aflatoxins B1, B2, G1, and G2 in olive oil, peanut oil, and sesame oil using immunoaffinity column cleanup, postcolumn derivatization, and liquid chromatography/fluorescence detection: collaborative study.

作者信息

Bao Lei, Liang Chengzhu, Trucksess Mary W, Xu Yanli, Lv Ning, Wu Zhenxing, Jing Ping, Fry Fred S

机构信息

Shandong Exit-Entry Inspection and Quarantine Bureau of China, Qing Dao, China 266002.

出版信息

J AOAC Int. 2012 Nov-Dec;95(6):1689-700. doi: 10.5740/jaoacint.12-199.

DOI:10.5740/jaoacint.12-199

PMID:23451385

Abstract

The accuracy, repeatability, and reproducibility characteristics of a method using immunoaffinity column (IAC) cleanup with postcolumn derivatization and LC with a fluorescence detector (FLD) for determination of aflatoxins (AFs; sum of AFs B1, B2, G1, and G2) in olive oil, peanut oil, and sesame oil have been established in a collaborative study involving 15 laboratories from six countries. Blind duplicate samples of blank, spiked at levels ranging from 0.25 to 20.0 microg/kg for AF, were analyzed. A naturally contaminated peanut oil sample was also included. Test samples were extracted with methanol-water (55 + 45, v/v). After shaking and centrifuging, the lower layer was filtered, diluted with water, and filtered through glass microfiber filter paper. The filtrate was then passed through an IAC, and the toxins were eluted with methanol. The toxins were then subjected to RPLC-FLD analysis after postcolumn derivatization. Average recoveries of AFs from olive oil, peanut oil, and sesame oil ranged from 84 to 92% (at spiking levels ranging from 2.0 to 20.0 microg/kg); of AFB1 from 86 to 93% (at spiking levels ranging from 1.0 to 10.0 microg/kg); of AFB2 from 89 to 95% (at spiking levels ranging from 0.25 to 2.5 microg/kg); of AFG1 from 85 to 97% (at spiking levels ranging from 0.5 to 5.0 microg/kg); and of AFG2 from 76 to 85% (at spiking levels ranging from 0.25 to 2.5 microg/kg). RSDs for within-laboratory repeatability (RSD(r)) ranged from 3.4 to 10.2% for AF, from 3.5 to 10.9% for AFB1, from 3.2 to 9.5% for AFB2, from 6.5 to 14.9% for AFG1, and from 4.8 to 14.2% for AFG2. RSDs for between-laboratory reproducibility (RSDR) ranged from 6.1 to 14.5% for AF, from 7.5 to 15.4% for AFB1, from 7.1 to 14.6% for AFB2, from 10.8 to 18.1% for AFG1, and from 7.6 to 23.7% for AFG2. Horwitz ratio values were < or = 2 for the analytes in the three matrixes.

摘要

在一项由来自六个国家的15个实验室参与的协作研究中，建立了一种使用免疫亲和柱（IAC）净化、柱后衍生化和带荧光检测器（FLD）的液相色谱法测定橄榄油、花生油和芝麻油中黄曲霉毒素（AFs；AFs B1、B2、G1和G2的总和）的方法的准确性、重复性和再现性特征。分析了AF添加水平为0.25至20.0微克/千克的盲法双份空白加标样品。还包括一份天然污染的花生油样品。测试样品用甲醇 - 水（55 + 45，v/v）萃取。振荡和离心后，下层过滤，用水稀释，再通过玻璃微纤维滤纸过滤。滤液然后通过IAC，毒素用甲醇洗脱。毒素在柱后衍生化后进行反相高效液相色谱 - 荧光检测分析。橄榄油、花生油和芝麻油中AFs的平均回收率为84%至92%（加标水平为2.0至20.0微克/千克）；AFB1为86%至93%（加标水平为1.0至10.0微克/千克）；AFB2为89%至95%（加标水平为0.25至2.5微克/千克）；AFG1为85%至97%（加标水平为0.5至5.0微克/千克）；AFG2为76%至85%（加标水平为0.25至2.5微克/千克）。实验室内重复性（RSD(r)）的相对标准偏差对于AF为3.4%至10.2%，对于AFB1为3.5%至10.9%，对于AFB2为3.2%至9.5%，对于AFG1为6.5%至14.9%，对于AFG2为4.8%至14.2%。实验室间再现性（RSDR）的相对标准偏差对于AF为6.1%至14.5%，对于AFB1为7.5%至15.4%，对于AFB2为7.1%至14.6%，对于AFG1为10.8%至18.1%，对于AFG2为7.6%至23.7%。三种基质中分析物的霍维茨比值≤2。