建立一种高效液相色谱-串联质谱法测定羽扇豆中生物碱的方法。

Development of an HPLC-MS/MS Method for the Determination of Alkaloids in Lupins.

机构信息

Department of Bioscience and Technology for Food, Agriculture and Environment, University of Teramo, Via Renato Balzarini 1, 64100 Teramo, Italy.

Department of Chemistry, Sapienza University of Rome, Piazzale Aldo Moro 5, 00185 Rome, Italy.

出版信息

Molecules. 2023 Feb 5;28(4):1531. doi: 10.3390/molecules28041531.

Lupin alkaloids (LAs) represent a class of toxic secondary metabolites in plants, in particular in spp.; they are produced as a defense mechanism due to their strong bitter taste and are very dangerous for human and animals. In this work, a sensitive and reliable high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analytical method for the identification and quantification of thirteen lupin alkaloids was developed and validated according to FDA guidelines. Efficient extraction and clean-up steps, carried out by solid-phase extraction, were finely tuned on the basis of the characteristics of the analytes and lupin samples, providing good selectivity with minimized matrix interference. The effectiveness of the method was proven by the satisfactory recovery values obtained for most of the analytes and a matrix effect ≤23% for all tested levels. In addition, a sensitive and reliable determination of the target compounds was obtained; LOQs were between 1 and 25 µg Kg, i.e., below the requested maximum levels (<200 mg Kg). The method was applied to evaluate the LAs profile in different batches of raw L. samples, varying in size and across farming treatments.

羽扇豆生物碱（LAs）是植物中一类有毒的次生代谢物，特别是在 spp. 中；它们是作为一种防御机制产生的，因为它们的强烈苦味，对人类和动物非常危险。在这项工作中，根据 FDA 指南，开发并验证了一种用于鉴定和定量十三种羽扇豆生物碱的灵敏可靠的高效液相色谱-串联质谱（HPLC-MS/MS）分析方法。基于分析物和羽扇豆样品的特性，对高效提取和净化步骤（固相萃取）进行了精细调整，从而具有良好的选择性，最大限度地减少了基质干扰。该方法通过获得大多数分析物令人满意的回收率值以及所有测试水平的基质效应均≤23%，证明了其有效性。此外，还获得了对目标化合物的灵敏和可靠的测定；LOQs 在 1 至 25 µg Kg 之间，即低于要求的最大水平（<200 mg Kg）。该方法应用于评估不同批次的生羽扇豆 L. 样品中的 LAs 分布，大小和种植处理各不相同。