Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, David Keir Building, Stranmillis Road, Belfast, BT9 5AG, UK.
Anal Bioanal Chem. 2014 Jan;406(3):757-70. doi: 10.1007/s00216-013-7488-7. Epub 2013 Dec 11.
Pyrrolizidine alkaloids (PAs) are a group of plant secondary metabolites with carcinogenic and hepatotoxic properties. When PA-producing plants contaminate crops, toxins can be transferred through the food chain and cause illness in humans and animals, most notably hepatic veno-occlusive disease. Honey has been identified as a direct risk of human exposure. The European Food Safety Authority has recently identified four groups of PAs that are of particular importance for food and feed: senecionine-type, lycopsamine-type, heliotrine-type and monocrotaline-type. Liquid or gas chromatography methods are currently used to detect PAs but there are no rapid screening assays available commercially. Therefore, the aim of this study was to develop a rapid multiplex ELISA test for the representatives of three groups of alkaloids (senecionine, lycopsamine and heliotrine types) that would be used as a risk-management tool for the screening of these toxic compounds in food and feed. The method was validated for honey and feed matrices and was demonstrated to have a detection capability less than 25 μg/kg for jacobine, lycopsamine, heliotrine and senecionine. The zinc reduction step introduced to the extraction procedure allows for the additional detection of the presence of N-oxides of PAs. This first multiplex immunoassay for PA detection with N-oxide reduction can be used for the simultaneous screening of 21 samples for >12 PA analytes. Honey samples (n = 146) from various origins were analysed for PA determination. Six samples were determined to contain measurable PAs >25 μg/kg by ELISA which correlated to >10 μg/kg by LC-MS/MS.
吡咯里西啶生物碱（PA）是一组具有致癌和肝毒性的植物次生代谢物。当产生 PA 的植物污染农作物时，毒素可以通过食物链转移，导致人类和动物患病，尤其是肝静脉闭塞性疾病。蜂蜜已被确定为人类接触的直接风险。最近，欧洲食品安全局确定了四类对食品和饲料特别重要的 PA：山金车型、藜芦碱型、千里光型和野百合碱型。目前使用液相或气相色谱法来检测 PA，但没有商业上可用的快速筛选分析方法。因此，本研究旨在开发一种用于三种生物碱（山金车型、藜芦碱型和千里光型）代表物的快速多重 ELISA 测试，该测试将作为筛选食品和饲料中这些有毒化合物的风险管理工具。该方法已针对蜂蜜和饲料基质进行了验证，并且在检测雅各宾碱、藜芦碱、千里光碱和山金车碱的能力方面，检测限小于 25μg/kg。在提取过程中引入的锌还原步骤可以额外检测到 PA 的 N-氧化物。这是首次具有 N-氧化物还原功能的 PA 多重免疫检测法，可用于同时筛选 21 个样本中的>12 种 PA 分析物。对来自不同来源的 146 份蜂蜜样品进行了 PA 测定。通过 ELISA 测定有 6 个样品中含有可测量的>25μg/kg 的 PA，与 LC-MS/MS 测定的>10μg/kg 相吻合。
