A method for concurrent diazomethane synthesis and substrate methylation in a 96-sample format
- 9 September 2010
- journal article
- Published by Springer Science and Business Media LLC in Nature Protocols
- Vol. 5 (10), 1619-1626
- https://doi.org/10.1038/nprot.2010.119
Abstract
In the emerging field of metabolomics, there is an increasing need for improving sample derivatization reactions for gas chromatographic-mass spectral analysis of metabolites with large numbers of samples. This protocol details the safe direct derivatization of organic acids using diazomethane in a 96-sample format. Diazomethane is a highly reactive gas that readily forms methyl esters with carboxylic functionalities, with minimal side products or nonvolatile reaction residues. However, diazomethane's reactivity and explosive potential make it hazardous to store and work with. In this procedure, diazomethane is generated in situ and used concurrently to methylate up to 96 samples simultaneously, thus reducing concerns about reagent stability and obviating the need for storage of solutions of the highly reactive gas. Once the diazomethane generator has been assembled, processing 96 samples takes 2-3 h using this procedure.Keywords
This publication has 16 references indexed in Scilit:
- A high-throughput method for the quantitative analysis of auxinsNature Protocols, 2010
- Extending the breadth of metabolite profiling by gas chromatography coupled to mass spectrometryTrAC Trends in Analytical Chemistry, 2008
- A high-throughput method for the quantitative analysis of indole-3-acetic acid and other auxins from plant tissueAnalytical Biochemistry, 2008
- Metabolome analysis by capillary electrophoresis–mass spectrometryJournal of Chromatography A, 2007
- Chemical derivatization and mass spectral libraries in metabolic profiling by GC/MS and LC/MS/MSJournal of Experimental Botany, 2004
- Development of a Continuous Process for the Industrial Generation of Diazomethane1Organic Process Research & Development, 2002
- Recent advances in non-silylation derivatization techniques for gas chromatographyJournal of Chromatography A, 1999
- Study of the reaction of diazomethane with dansylated amino acid derivatives by gas chromatography/mass spectrometryJournal of Mass Spectrometry, 1990
- Convenient apparatus for the generation of small amounts of diazomethaneJournal of Chromatography A, 1984
- Esterification of Fatty Acids with Diazomethane on a Small ScaleAnalytical Chemistry, 1960