Background

Chemical derivatization is a common technique in liquid chromatography-mass spectrometry (LC-MS) metabolomics used to improve the ionizability and chromatographic properties of metabolites in complex biological samples. This process facilitates better detection and separation of a wide array of compounds. The reagent 2-(4-boronobenzyl) isoquinolin-2-ium bromide (BBII), developed as a glucose labeling reagent for matrix-assisted laser desorption/ionization MS, enhances ionization for glucose and other hydroxyl metabolites. Its quaternary ammonium group increases ionization efficiency, and its rapid reaction time simplifies pretreatment procedures.

Results

We developed a novel post-column derivatization (PCD) method using BBII to boost the detection sensitivity of hydroxyl metabolites in LC-MS. By optimizing this BBII PCD approach with 14 hydroxyl-containing compounds, we were able to detect previously undetectable metabolites such as glucose, ribose, and long-chain alcohols. Sensitivity enhancements for these metabolites ranged from 1.1 to 42.9-fold. Applying this method to metabolic profiling of hydroxyl metabolites in the DBTRG-05MG glioblastoma cell line, with and without treatment with the new drug MFB [1-(4-chlorobenzyl)-2-(5-methyl-2-furfurylideneamino)benzimidazole], revealed several hydroxyl metabolites with significantly reduced levels post-treatment.

Significance and Novelty

This study presents a new BBII PCD method that substantially improves the detection sensitivity of hydroxyl metabolites in LC-MS. This innovative approach is highly valuable for untargeted metabolomics studies in biological and clinical research, offering a robust tool for identifying metabolite changes and advancing our understanding of metabolic processes in disease and therapeutic contexts.

中文翻译：

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一种用于增强羟基代谢物检测的灵敏柱后衍生化方法

 背景

化学衍生化是液相色谱-质谱 （LC-MS） 代谢组学中的一种常用技术，用于改善复杂生物样品中代谢物的电离性和色谱特性。该工艺有助于更好地检测和分离各种化合物。试剂 2-（4-硼苄基）异喹啉-2-溴化铵 （BBII） 作为基质辅助激光解吸/电离 MS 的葡萄糖标记试剂开发，可增强葡萄糖和其他羟基代谢物的电离。其季铵基团提高了电离效率，其快速反应时间简化了预处理程序。

 结果

我们开发了一种使用 BBII 的新型柱后衍生化 （PCD） 方法，以提高 LC-MS 中羟基代谢物的检测灵敏度。通过使用 14 种含羟基化合物优化这种 BBII PCD 方法，我们能够检测以前无法检测到的代谢物，例如葡萄糖、核糖和长链醇。这些代谢物的灵敏度增强范围为 1.1 至 42.9 倍。将该方法应用于 DBTRG-05MG 胶质母细胞瘤细胞系中羟基代谢物的代谢谱分析，有和没有用新药 MFB [1-（4-氯苄基）-2-（5-甲基-2-糠酰亚氨基）苯并咪唑]处理，揭示了几种羟基代谢物在处理后水平显着降低。

 意义和新颖性

本研究提出了一种新的 BBII PCD 方法，该方法显著提高了 LC-MS 中羟基代谢物的检测灵敏度。这种创新方法对于生物学和临床研究中的非靶向代谢组学研究非常有价值，为识别代谢物变化和促进我们对疾病和治疗环境中代谢过程的理解提供了强大的工具。