We have developed and validated a novel LC-MS/MS method for simultaneously analyzing amino acids, biogenic amines, and their acetylated and methylated derivatives in plants. This method involves a one-step extraction of 2-5 mg of lyophilized plant material followed by fractionation of different biogenic amine forms and exploits an efficient combination of hydrophilic interaction chromatography (HILIC), reversed phase (RP) chromatography with pre-column derivatization, and tandem mass spectrometry. This approach enables high-throughput processing of plant samples, significantly reducing the time needed for analysis and its cost. We also present a new synthetic route for deuterium-labelled polyamines. The LC-MS/MS method was rigorously validated by quantifying levels of nitrogen-related metabolites in seedlings of seven plant species including Arabidopsis, maize, and barley, all of which are commonly used model organisms in plant science research. Our results revealed substantial variations in the abundance of these metabolites between species, developmental stages, and growth conditions, particularly for the acetylated and methylated derivatives and the various polyamine fractions. However, the biological relevance of these plant metabolites is currently unclear. Overall, this work contributes significantly to the field of plant science by providing a powerful analytical tool and setting the stage for future investigations into the functions of these nitrogen-related metabolites in plants.

中文翻译：

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氮相关植物代谢物的综合 LC-MS/MS 分析

我们开发并验证了一种新型 LC-MS/MS 方法，用于同时分析植物中的氨基酸、生物胺及其乙酰化和甲基化衍生物。该方法涉及一步提取 2-5 毫克冻干植物材料，然后分馏不同的生物胺形式，并利用亲水相互作用色谱 (HILIC)、反相 (RP) 色谱与柱前衍生化的有效组合，和串联质谱分析。这种方法可以对植物样品进行高通量处理，从而显着减少分析所需的时间和成本。我们还提出了氘标记多胺的新合成路线。通过量化拟南芥、玉米和大麦等七种植物幼苗中氮相关代谢物的水平，对 LC-MS/MS 方法进行了严格验证，所有这些植物都是植物科学研究中常用的模式生物。我们的结果揭示了这些代谢物的丰度在物种、发育阶段和生长条件之间存在显着差异，特别是乙酰化和甲基化衍生物以及各种多胺组分。然而，这些植物代谢物的生物学相关性目前尚不清楚。总的来说，这项工作为植物科学领域做出了重大贡献，提供了强大的分析工具，并为未来研究植物中这些氮相关代谢物的功能奠定了基础。