Improved stability of TMS derivatives for the robust quantification of plant polar metabolites by gas chromatography-mass spectrometry
Improved stability of TMS derivatives for the robust quantification of plant polar metabolites by gas chromatography-mass spectrometry
复制标题
DOI：
10.1016/j.jchromb.2014.08.040
复制
发表时间：
2014-11-01
影响因子：
3
通讯作者：
Pau-Roblot, Corinne
中科院分区：
文献类型：
--
作者：
Quero, Anthony;Jousse, Cyril;Pau-Roblot, Corinne
植物代谢物谱分析通常由甲氧基三甲基硅基（TMS）衍生物的GC-MS进行。这一技术是可靠的，可以对电子电离产生的光谱进行库搜索。然而，最近的文章描述了与一些TMS衍生物的低稳定性相关的问题。这限制了GC-MS用于需要大量定性和定量分析的代谢组学研究。本工作的目的是确定可以提高TMS衍生物稳定性的实验条件。这将有助于代谢组学方法所需的大规模实验设计的分析。为了获得良好的重复性，研究了分析过程中样品的取样条件和保存温度。从一个小瓶中多次注射一个样品导致高变化，而从不同的小瓶中注射一个样品改善了分析。然而，在注射前，一些氨基酸TMS衍生物在自动进样器的储存过程中被降解。衍生48 h后，仅检测到初始量的10%的谷氨酰胺3 TMS和谷氨酸3 TMS和66%的α -丙氨酸2 TMS。在4℃保存至注射前，所有TMS衍生物在12 h内保持稳定；在-20℃下，它们在72小时内保持稳定。根据所有这些结果的综合，提出了详细的分析方法。它能够对极性代谢物进行可靠的定量分析，有助于进一步使用GC-MS进行植物代谢组学研究。(C) 2014 Elsevier B.V.版权所有
Plant metabolite profiling is commonly carried out by GC-MS of methoximated trimethylsilyl (TMS) derivatives. This technique is robust and enables a library search for spectra produced by electron ionization. However, recent articles have described problems associated with the low stability of some TMS derivatives. This limits the use of GC-MS for metabolomic studies that need large sets of qualitative and quantitative analyses. The aim of this work is to determine the experimental conditions in which the stability of TMS derivatives could be improved. This would facilitate the analysis of the large-scale experimental designs needed in the metabolomics approach. For good repeatability, the sampling conditions and the storage temperature of samples during analysis were investigated. Multiple injections of one sample from one vial led to high variations while injection of one sample from different vials improved the analysis. However, before injection, some amino acid TMS derivatives were degraded during the storage of vials in the autosampler. Only 10% of the initial quantity of glutamine 3 TMS and glutamate 3 TMS and 66% of alpha-alanine 2 TMS was detected 48 h after derivatization. When stored at 4 degrees C until injection, all TMS derivatives remained stable for 12 h; at -20 degrees C, they remained stable for 72 h. From the integration of all these results, a detailed analytical procedure is thus proposed. It enables a robust quantification of polar metabolites, useful for further plant metabolomics studies using GC-MS. (C) 2014 Elsevier B.V. All rights reserved.