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2016•BioanalysisRequires access

Investigation of The Derivatization Conditions for GC–MS Metabolomics of Biological Samples

Georgios Moros, Anastasia Chrysovalantou Chatziioannou, Helen Gika, Νikolaos Raikos, Georgios Theodoridis

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Abstract

AIM: Metabolomics applications represent an emerging field where significant efforts are directed. Derivatization consists prerequisite for GC-MS metabolomics analysis. METHODS: Common silylation agents were tested for the derivatization of blood plasma. Optimization of methoxyamination and silylation reactions was performed on a mixture of reference standards, consisting of 46 different metabolites. Stability of derivatized metabolites was tested at 4°C. RESULTS: Optimum results were achieved using N-methyl-N-(trimethylsilyl)trifluoroacetamide. Methoxyamination at room temperature for 24 h followed by 2-h silylation at high temperature lead to efficient derivatization. CONCLUSION: Formation and stability of derivatives among metabolites differ greatly, so derivatization should be studied before application in metabolomics studies.

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AIM: Metabolomics applications represent an emerging field where significant efforts are directed. Derivatization consists prerequisite for GC-MS metabolomics analysis. METHODS: Common silylation agents were tested for the derivatization of blood plasma. Optimization of methoxyamination and silylation reactions was performed on a mixture of reference standards, consisting of 46 different metabolites. Stability of derivatized metabolites was tested at 4°C. RESULTS: Optimum results were achieved using N-methyl-N-(trimethylsilyl)trifluoroacetamide. Methoxyamination at room temperature for 24 h followed by 2-h silylation at high temperature lead to efficient derivatization. CONCLUSION: Formation and stability of derivatives among metabolites differ greatly, so derivatization should be studied before application in metabolomics studies.

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Available abstract

AIM: Metabolomics applications represent an emerging field where significant efforts are directed. Derivatization consists prerequisite for GC-MS metabolomics analysis. METHODS: Common silylation agents were tested for the derivatization of blood plasma. Optimization of methoxyamination and silylation reactions was performed on a mixture of reference standards, consisting of 46 different metabolites. Stability of derivatized metabolites was tested at 4°C. RESULTS: Optimum results were achieved using N-methyl-N-(trimethylsilyl)trifluoroacetamide. Methoxyamination at room temperature for 24 h followed by 2-h silylation at high temperature lead to efficient derivatization. CONCLUSION: Formation and stability of derivatives among metabolites differ greatly, so derivatization should be studied before application in metabolomics studies.

Key concepts: Derivatization, Silylation, Trimethylsilyl, Metabolomics, Chemistry, Chromatography, BSTFA, Gas chromatography–mass spectrometry

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