Department of Chemical and Physical Health Risks, Brussels, Belgium.
DNAGensee, Le Bourget Du Lac Cedex, France.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2021 Jun;38(6):962-977. doi: 10.1080/19440049.2021.1889043. Epub 2021 Mar 29.
Pyrrolizidine alkaloids (PAs) and their corresponding N-oxides (PANOs) are natural protoxins biosynthesised by many plant species and are responsible for occasional fatal intoxication outbreaks due to the consumption of contaminated food. However, only in rare cases has the origin of the contamination been determined. Although their presence has been studied in many matrices, occurrence data on popular widely used cooking herbs, seeds, spices and leafy vegetables is very scarce. Therefore, a systematic study on the occurrence of PA/PANOs contaminations in these popular herbal items, available on the Belgian market, was performed, by means of a validatedtargeted LC-MS/MS analysis, followed by multilocus DNA metabarcoding to track back the origin of the contamination for seven highly to moderately contaminated samples. Our results clearly indicate that 21% of the seed spices and 25% seed-based aromatic mixes contain an amount higher than 400 μg of the 30 summed targeted PAs and PANOs per kg, the value which is currently under discussion by the European member states to be set as a maximum threshold. For both the herbs and the herb-based mixes only 7% of analysed samples exceeded these levels. As a proof of concept, multilocus DNA metabarcoding was performed on six highly contaminated samples, belonging to each subtype matrix, containing high levels of heliotrine type of PA/PANOs. Each time the analysis demonstrated the presence of DNA from a plant species belonging to the plant genus . Moreover, a contaminated leafy vegetable sample, containing solely senecionine type PA/PANOs, contained DNA from . Taken together, it can be stated that the proposed combinatory chemical and molecular techniques could be used to verify if a PA/PANO(s) contamination occurred in these popular cooking items and to pinpoint the origin the contamination, which is pivotal in the case of a detrimental intoxication or intoxication outbreak.
吡咯里西啶生物碱（PAs）及其相应的 N-氧化物（PANOs）是许多植物物种生物合成的天然前毒素，由于食用受污染的食物，偶尔会导致致命的中毒爆发。然而，只有在极少数情况下确定了污染的来源。尽管已经在许多基质中研究了它们的存在，但关于在比利时市场上流行的烹饪香草、种子、香料和叶菜类蔬菜中 PA/PANO 污染的发生数据非常稀缺。因此，通过验证的靶向 LC-MS/MS 分析，对这些在比利时市场上流行的草药制品中的 PA/PANO 污染进行了系统研究，然后通过多基因 DNA 代谢组学对 7 个高度到中度污染的样本进行追溯污染来源。我们的研究结果清楚地表明，21%的种子香料和 25%的基于种子的芳香混合物中含有超过 400μg/kg 的 30 种目标 PA 和 PANO 之和，这一数值目前正在由欧洲成员国讨论，以设定为最大阈值。对于草药和基于草药的混合物，只有 7%的分析样本超过了这些水平。作为概念验证，对属于六份高度污染样本的 6 份高度污染样本进行了多基因 DNA 代谢组学分析，这六份样本都属于含有高水平海利毒素型 PA/PANO 的亚基质。每次分析都证明了属于植物属的植物种的 DNA 存在。此外，一份仅含有山莴苣型 PA/PANO 的污染叶菜类蔬菜样本中，含有 DNA 。综上所述，可以说，所提出的组合化学和分子技术可用于验证这些流行的烹饪制品是否发生了 PA/PANO 污染，并确定污染的来源，这在发生有害中毒或中毒爆发时至关重要。
