同时测定肉制品中的赭曲霉毒素 A、霉酚酸和伏马菌素 B(2)。
Simultaneous determination of ochratoxin A, mycophenolic acid and fumonisin B(2) in meat products.
机构信息
Department of Systems Biology, Technical University of Denmark, Building 221, 2800, Lyngby, Denmark.
出版信息
Anal Bioanal Chem. 2010 Oct;398(3):1535-42. doi: 10.1007/s00216-010-4059-z. Epub 2010 Aug 9.
Here we present a method for simultaneous determination of the fungal metabolites mycophenolic acid, ochratoxin A (OTA) and fumonisin B(2) (FB(2)) in meat products. Extraction was performed with water-acetonitrile, followed by acetone-induced precipitation of salts and proteins. Purification and identification of analytes was performed by mixed-mode reversed-phase anion-exchange chromatography in direct ion-exchange mode, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) detection. Quantification was based on isotope dilution with fully (13)C-labelled FB(2) and OTA, and matrix-spiked calibration curves. Fermented sausages inoculated with an OTA- and FB(2)-producing strain of Aspergillus niger were analysed, but no analytes were detected. Analysis of 22 retail products showed one Parma meat with a very high level of OTA contamination (56-158 microg/kg) that clearly exceeded the Italian regulatory limit of 1 microg/kg. This sample and uninfected control samples were subsequently reanalysed, and the high OTA content was verified by two other techniques: (i) LC-time-of-flight MS confirmed the accurate mass as well as chlorine isotope pattern; and (ii) sample methylation in methanol-BF(3) and subsequent LC-MS/MS provided indirect confirmation by detection of the OTA methyl ester. In the contaminated Parma ham, the high OTA level most likely originated from growth of Penicillium nordicum on the meat.
我们在此介绍一种用于同时测定肉类产品中真菌代谢产物麦考酚酸、赭曲霉毒素 A(OTA)和伏马菌素 B1(FB1)的方法。采用水-乙腈提取,然后用丙酮沉淀盐和蛋白质。通过混合模式反相阴离子交换色谱在直接离子交换模式下进行分析物的净化和鉴定,然后通过液相色谱-串联质谱(LC-MS/MS)检测。采用完全(13)C 标记的 FB1 和 OTA 进行同位素稀释和基质加标校准曲线进行定量。用产 OTA 和 FB1 的黑曲霉菌株接种发酵香肠进行分析,但未检测到分析物。对 22 个零售产品进行分析,发现一片帕尔马火腿受到 OTA 严重污染(56-158μg/kg),明显超过意大利监管限量 1μg/kg。对该样品和未感染对照样品进行了重新分析,通过另外两种技术验证了 OTA 的高含量:(i)LC-飞行时间 MS 准确确认了质量和氯同位素模式;(ii)甲醇-BF3 中的样品甲基化,然后通过 LC-MS/MS 检测 OTA 甲酯,提供了间接确认。在受污染的帕尔马火腿中,高 OTA 水平很可能来自肉上的青霉属北菌的生长。