Quantification of Amine- and Alcohol-Containing Metabolites in Saline Samples Using Pre-extraction Benzoyl Chloride Derivatization and Ultrahigh Performance Liquid Chromatography Tandem Mass Spectrometry (UHPLC MS/MS)
Quantification of Amine- and Alcohol-Containing Metabolites in Saline Samples Using Pre-extraction Benzoyl Chloride Derivatization and Ultrahigh Performance Liquid Chromatography Tandem Mass Spectrometry (UHPLC MS/MS)
复制标题
使用预提取苯甲酰氯衍生化和超高效液相色谱串联质谱 (UHPLC MS/MS) 定量盐水样品中的胺和醇代谢物
DOI：
10.1021/acs.analchem.0c03769
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发表时间：
2021
影响因子：
7.4
通讯作者：
Kujawinski, Elizabeth B.
中科院分区：
文献类型：
--
作者：
Widner, Brittany;Kido Soule, Melissa C.;Ferrer-González, Frank Xavier;Moran, Mary Ann;Kujawinski, Elizabeth B.
溶解的代谢产物为微生物系统提供营养、能量和化学信号。然而，海洋系统中这些过程的全部范围和程度尚不清楚，主要是由于方法不足，包括使用普通固相萃取树脂对小的极性化合物的萃取效果不佳。在这里，我们利用提取前衍生化和高效液相色谱电喷雾电离串联质谱（UHPLC-ESI-MS/MS）来检测和定量海水和盐水培养基中的目标溶解代谢物。通过伯胺、仲胺和醇官能团，用苯甲酰氯衍生化二甲双胍，并使用由13 C6标记的苯甲酰氯制备的稳定同位素标记的内标物（SIL-IS）进行定量。我们优化了衍生化，提取和现场和文化样品的样品制备和评估基质衍生的偏差。我们优化了73种常见代谢物的定量方法，其中50种由于提取效率低而无法在不衍生化的情况下定量。在73种代谢物中，66种在培养基或海水中被鉴定，其中45种被定量。该衍生化方法灵敏（检测限= pM至nM）、快速（每个样品约105分钟）和高通量。
Dissolved metabolites serve as nutrition, energy, and chemical signals for microbial systems. However, the full scope and magnitude of these processes in marine systems are unknown, largely due to insufficient methods, including poor extraction of small, polar compounds using common solid-phase extraction resins. Here, we utilized pre-extraction derivatization and ultrahigh performance liquid chromatography electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) to detect and quantify targeted dissolved metabolites in seawater and saline culture media. Metabolites were derivatized with benzoyl chloride by their primary and secondary amine and alcohol functionalities and quantified using stable isotope-labeled internal standards (SIL-ISs) produced from13C6-labeled benzoyl chloride. We optimized derivatization, extraction, and sample preparation for field and culture samples and evaluated matrix-derived biases. We have optimized this quantitative method for 73 common metabolites, of which 50 cannot be quantified without derivatization due to low extraction efficiencies. Of the 73 metabolites, 66 were identified in either culture media or seawater and 45 of those were quantified. This derivatization method is sensitive (detection limits = pM to nM), rapid (∼5 min per sample), and high throughput.