Department of Chemistry, DBA Center, ETSEA, University of Lleida, 25003, Lleida, Spain.
Scientific Technical Service TCEM, University of Lleida, 25003, Lleida, Spain.
Metabolomics. 2021 Feb 6;17(2):22. doi: 10.1007/s11306-021-01771-w.
The metabolomic profile is an essential tool for understanding the physiological processes of biological samples and their changes. In addition, it makes it possible to find new substances with industrial applications or use as drugs. As GC-MS is a very common tool for obtaining the metabolomic profile, a simple and fast method for sample preparation is required.
The aim of this research was to develop a direct derivatization method for GC-MS to simplify the sample preparation process and apply it to a wide range of samples for non-targeted metabolomic analysis purposes.
One pot combined esterification of carboxylic acids with methanol and silylation of the hydroxyl groups was achieved using a molar excess of chlorotrimethylsilane with respect to methanol in the presence of pyridine.
The metabolome profile obtained from different samples, such as bilberry and cherry cuticles, olive leaves, P. aeruginosa and E. coli bacteria, A. niger fungi and human sebum from the ceruminous gland, shows that the procedure allows the identification of a wide variety of metabolites. Aliphatic fatty acids, hydroxyfatty acids, phenolic and other aromatic compounds, fatty alcohols, fatty aldehydes dimethylacetals, hydrocarbons, terpenoids, sterols and carbohydrates were identified at different MSI levels using their mass spectra.
The metabolomic profile of different biological samples can be easily obtained by GC-MS using an efficient simultaneous esterification-silylation reaction. The derivatization method can be carried out in a short time in the same injection vial with a small amount of reagents.
代谢组学谱是理解生物样本生理过程及其变化的重要工具。此外，它还可以发现具有工业应用或用作药物的新物质。由于 GC-MS 是获取代谢组学谱的常用工具，因此需要一种简单、快速的样品制备方法。
本研究旨在开发一种 GC-MS 的直接衍生化方法，以简化样品制备过程，并将其应用于广泛的样品，用于非靶向代谢组学分析。
在吡啶存在下，使用相对于甲醇的摩尔过量的三甲基氯硅烷，实现了羧酸与甲醇的一锅法联合酯化和羟基的硅烷化。
从不同样品（如越橘和樱桃表皮、橄榄叶、铜绿假单胞菌和大肠杆菌细菌、黑曲霉真菌和人耳垢中的皮脂）获得的代谢组学谱表明，该方法允许鉴定出多种代谢物。使用其质谱图，在不同的 MSI 水平上鉴定了脂族脂肪酸、羟基脂肪酸、酚类和其他芳香族化合物、脂肪醇、脂肪醛二甲缩醛、烃类、萜类、甾醇和碳水化合物。
使用高效的同时酯化-硅烷化反应，通过 GC-MS 可以轻松获得不同生物样本的代谢组学谱。衍生化方法可以在少量试剂的同一个进样瓶中短时间内进行。
