Analysis of pyrrolizidine alkaloids in food – determination of the sum by high-performance thin-layer chromatography and identi-fication by planar solid phase extraction coupled with mass spectrometry

应开发两种分析替代品，作为食品中吡咯里西丁生物碱(PA)分析的筛选方法。通过简便的方法，可以测定样品中PA的总含量(总和参数)，并可以快速、经济有效地鉴定和定量单个PA。在此背景下，应从根本上研究和认识降低PA。此外，平面固相萃取-质谱仪(PSPE-MS)的液基联用应自动用于PA的分析，并为常规应用做好准备。此外，离子迁移率将在pSPE-MS分离等压PA的方法中进行研究。在用高效液相色谱-荧光检测法(HPTLC-FLD)分析总PA时，PA被还原到甘氨酸碱基上，然后衍生化成荧光产物。这一方法将适用于反式和贺利曲定类型的PA，并将覆盖PA的90%。用pSPE-MS测定完整PA的方法，在MS检测前使用直接在线偶联而不需要层析分离，将以一种非常创新的方式提供单一质谱中所有PA的鉴定和定量，因为只需要在依赖于数据库的评估方法中进行几公里。在第一步中，将开发和优化具有代表性的PA的HPTLC-FLD和PSPE-MS筛选方法，并随后应用于各种茶叶基质。在接下来的步骤中，这两种筛查方法都将适用于经常受到PA污染的食品，如香料、蜂蜜和谷类产品。不同的食物基质构成了巨大的挑战，因为与PA含量相比，基质负荷很大，而且基质非常复杂。高效和经济高效的筛选方法将为PA含量提供可靠的结果，从而提供关于PA污染食品类别的必要数据。此外，这两种方法的结合将代表一种非常有效的分析策略。这两种方法都将迅速提供关于苯丙酸污染的数据，因此，对于估计接触苯丙酸非常有价值，这是欧洲联盟迫切需要和声称的。此外，还可以对最近设置的最高水平进行监控。该项目的研究成果还将为自动化液基PSPE-MS联用的基础研究服务，为联用策略应用于各种后续项目提供依据，并为实际分析问题提供创新的解决方案。因此，几种易于执行和高效的筛选方法的开发将是有保证的。因此，所获得的结果和知识不仅将有助于PA的分析，而且将为现代食品分析提供基础和广泛的帮助。

Two analytical alternatives as screening methods for the analysis of pyrrolizidine alkaloids (PA) in food should be developed. By the easy to perform methods, the determination of the total PA content (sum parameter) in the sample and a fast and cost-effective identification and quantitation of single PA should be possible. In this context, the reduction of the PA should be fundamentally studied and understand. In addition, the liquid-based coupling of planar solid phase extraction with mass spectrometry (pSPE–MS) should be automated for the analysis of PA and completely elaborated for routine application. Furthermore, ion mobility will be investigated within the pSPE–MS approach for the separation of isobaric PA. For the analysis of the total PA by high-performance liquid chromatography with fluorescent detection (HPTLC–FLD), PA will be reduced to the necine base and derivatized to a fluorescent product. This approach will be applicable to PA of the retronecine and heliotridine type and will cover > 90% of the PA. The method for the determination of intact PA by pSPE–MS, which uses the direct online coupling without a chromatographic separation before MS detection, will offer the identification and quantitation of all PA in a single mass spectrum in a very innovative way, since only a few klicks within a database-dependent evaluation approach will be necessary. In the first step, the HPTLC–FLD and the pSPE–MS screening methods will be developed and optimized for representative PA, and subsequently, applied to various tea matrices. In following steps, both screening methods will be adapted to food that is frequently contaminated with PA, such as spices, honey, and cereal-based products. The different food matrices pose a great challenge, because the matrix load is immense compared to the PA content, and the matrices are highly complex. The efficient and cost-effective screening methods will deliver reliable results for the PA content, and thus, provide necessary data on the contamination of food categories with PA. Furthermore, the combination of both methods will represent a very efficient analysis strategy. Both methods will rapidly deliver data on the contamination with PA and, therefore, are very valuable for the estimation of the exposure to PA, which is urgently needed and claimed by the European Union. Additionally, monitoring of recently set maximum levels will be possible. The results of the project will moreover serve the fundamental research on the automated liquid-based pSPE–MS coupling, providing a basis for the coupling strategy to be applicable to various further projects, and offer innovative solutions for actual analytical questions. Thus, the development of several easy to perform and efficient screening methods will be guaranteed. Consequently, the obtained results and knowledge will not only be useful for the PA analysis, but provide fundamental and broad benefit for modern food analysis.