Acylcarnitines (ACs) play important roles in the fatty acid β-oxidation and are considered as diagnostic markers for many diseases. Accurate determination of ACs remains challenging due to their low abundance, high structure diversity, and limited availability of standard compounds. In this study, microwave-assisted Tmt-PP (p-[3,5-(dimethylamino)-2,4,6-triazine] benzene-1-sulfonyl piperazine) derivatization was utilized to facilitate the liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS) determination of ACs. The result indicated that Tmt-PP labeling enables the prediction of the retention time and MS response of ACs and enhances their MS response up to 4 times. The introduction of the microwave during the derivatization procedure greatly improved the reaction efficiency, demonstrated by the shortened reaction time from 90 to 1 min. Furthermore, we applied a strategy named quantitative analysis of multi-components by a single marker (QAMS) for the assay of 26 ACs with only 5 AC standards, solving the standard availability issue to a large extent. The established workflow was applied to discover dysregulated ACs in xenograft colon cancer mice, and the quantification results were highly comparable with traditional methods where there were the corresponding standards for each AC. Our study demonstrated that chemical derivatization-based LC–MS/MS integrated with the QAMS strategy is robust for the identification and quantification of ACs and has great potential in targeted metabolomics study.

中文翻译：

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使用基于集成 Tmt-PP 衍生化的 LC-MS/MS 对酰基肉碱进行绝对定量，并通过单一标记策略对多组分进行定量分析

酰基肉碱（ACs）在脂肪酸β-氧化中起重要作用，被认为是许多疾病的诊断标志物。由于 AC 丰度低、结构多样性高以及标准化合物的可用性有限，准确测定 AC 仍然具有挑战性。在本研究中，微波辅助 Tmt-PP ( p-[3,5-(二甲氨基)-2,4,6-三嗪]苯-1-磺酰基哌嗪)衍生化用于促进液相色谱与串联质谱联用 (LC-MS/MS) 测定活性炭。结果表明，Tmt-PP 标记能够预测 ACs 的保留时间和 MS 响应，并将其 MS 响应提高 4 倍。在衍生化过程中引入微波极大地提高了反应效率，反应时间从 90 分钟缩短到 1 分钟就证明了这一点。此外，我们应用了一种名为通过单一标记物进行多组分定量分析 (QAMS) 的策略，用于仅 5 个 AC 标准的 26 个 AC 的测定，在很大程度上解决了标准可用性问题。已建立的工作流程用于发现异种移植结肠癌小鼠中失调的 AC，定量结果与传统方法具有高度可比性，传统方法每个 AC 都有相应的标准。我们的研究表明，基于化学衍生化的 LC-MS/MS 与 QAMS 策略相结合，对于 AC 的鉴定和定量非常有效，并且在靶向代谢组学研究中具有巨大潜力。