title: Improved stability of TMS derivatives for the robust quantification of plant polar metabolites by gas chromatography-mass spectrometry.
abstract: 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 α-alanine 2 TMS was detected 48 h after derivatization. When stored at 4 °C until injection, all TMS derivatives remained stable for 12 h; at -20 °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.
authors.affiliation: Unité de Biologie des Plantes et Innovation, Université de Picardie Jules Verne, IUT d'Amiens, Dept. GB, Avenue des Facultés, Le Bailly, 80025 Amiens Cedex, France.
authors.forename: Anthony
authors.initials: A
authors.lastname: Quéro
authors.affiliation: Unité de Biologie des Plantes et Innovation, Université de Picardie Jules Verne, 33 rue Saint Leu, 80039 Amiens Cedex, France.
authors.forename: Cyril
authors.initials: C
authors.lastname: Jousse
authors.affiliation: Unité de Biologie des Plantes et Innovation, Université de Picardie Jules Verne, 33 rue Saint Leu, 80039 Amiens Cedex, France.
authors.forename: Michelle
authors.initials: M
authors.lastname: Lequart-Pillon
authors.affiliation: Unité de Biologie des Plantes et Innovation, Université de Picardie Jules Verne, 33 rue Saint Leu, 80039 Amiens Cedex, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Gontier
authors.affiliation: Laboulet Semences S.A., 1 rue Carnot, 80270 Airaines, France.
authors.forename: Xavier
authors.initials: X
authors.lastname: Guillot
authors.affiliation: Unité de Biologie des Plantes et Innovation, Université de Picardie Jules Verne, IUT d'Amiens, Dept. GB, Avenue des Facultés, Le Bailly, 80025 Amiens Cedex, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Courtois
authors.affiliation: Unité de Biologie des Plantes et Innovation, Université de Picardie Jules Verne, IUT d'Amiens, Dept. GB, Avenue des Facultés, Le Bailly, 80025 Amiens Cedex, France.
authors.forename: Josiane
authors.initials: J
authors.lastname: Courtois
authors.affiliation: Unité de Biologie des Plantes et Innovation, Université de Picardie Jules Verne, 33 rue Saint Leu, 80039 Amiens Cedex, France. Electronic address: corinne.pau-roblot@u-picardie.fr.
authors.forename: Corinne
authors.initials: C
authors.lastname: Pau-Roblot
mesh_terms: D000596:Amino Acids / Q000032:analysis / Q000737:chemistry; D017360:Arabidopsis / Q000378:metabolism; D002241:Carbohydrates / Q000032:analysis / Q000737:chemistry; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D055442:Metabolome / Q000502:physiology*; D055432:Metabolomics / Q000379:methods*; D012639:Seeds / Q000737:chemistry; D013696:Temperature; D014297:Trimethylsilyl Compounds / Q000032:analysis* / Q000737:chemistry
chemical_list: D000596:Amino Acids; D002241:Carbohydrates; D014297:Trimethylsilyl Compounds
keywords: Amino acids; GC–MS; Metabolite profiling; TMS derivatives