Optimization of a GC-MS Injection-Port Derivatization Methodology to Enhance Metabolomics Analysis Throughput in Biological Samples

• Published in: Journal of proteome research 2022-11, Vol.21 (11), p.2555-2565
• Main Authors: Foguet-Romero, Elisabet, Samarra, Iris, Guirro, Maria, Riu, Marc, Joven, Jorge, Menendez, Javier A., Canela, Núria, DelPino-Rius, Antoni, Fernández-Arroyo, Salvador, Herrero, Pol
• Format: Article
• Language: English
• Subjects: Acids; Amino Acids; Gas Chromatography-Mass Spectrometry - methods; Humans; Indicators and Reagents; Metabolomics - methods
• ISSN: 1535-3893; 1535-3907
• DOI: 10.1021/acs.jproteome.2c00119

Advances in metabolomics analysis and data treatment increase the knowledge of complex biological systems. One of the most used methodologies is gas chromatography-mass spectrometry (GC-MS) due to its robustness, high separation efficiency, and reliable peak identification through curated databases. However, methodologies are not standardized, and the derivatization steps in GC-MS can introduce experimental errors and take considerable time, exposing the samples to degradation. Here, we propose the injection-port derivatization (IPD) methodology to increase the throughput in plasma metabolomics analysis by GC-MS. The IPD method was evaluated and optimized for different families of metabolites (organic acids, amino acids, fatty acids, sugars, sugar phosphates, etc.) in terms of residence time, injection-port temperature, and sample/derivatization reagent ratio. Finally, the method’s usefulness was validated in a study consisting of a cohort of obese patients with or without nonalcoholic steatohepatitis. Our results show a fast, reproducible, precise, and reliable method for the analysis of biological samples by GC-MS. Raw data are publicly available at MetaboLights with Study Identifier MTBLS5151.