Chemical identification strategies using liquid chromatography-photodiode array-solid-phase extraction-nuclear magnetic resonance/mass spectrometry

The identification of metabolites in biochemical studies is a major bottleneck in the proliferating field of metabolomics. In particular in plant metabolomics, given the diversity and abundance of endogenous secondary metabolites in plants, the identification of these is not only challenging but also essential to understanding their biological role in the plant, and their value to quality and nutritional attributes as food crops. With the new generation of analytical technologies, in which liquid chromatography (LC)-mass spectrometry (MS) and nuclear magnetic resonance (NMR) play a pioneering role, profiling metabolites in complex extracts is feasible at high throughput. However, the identification of key metabolites remains a limitation given the analytical effort necessary for traditional structural elucidation strategies. The hyphenation of LC-solid phase extraction (SPE)-NMR is a powerful analytical platform for isolating and concentrating metabolites for unequivocal identification by NMR measurements. The combination with LC-MS is a relatively straightforward approach to obtaining all necessary information for structural elucidation. Using this set-up, we could, as an example, readily identify five related glycosylated phenolic acids present in broccoli (Brassica oleracea, group Italica, cv Monaco): 1,2-di-O-E-sinapoyl-β-gentiobiose, 1-O-E-sinapoyl-2-O-E-feruloyl-β-gentiobiose, 1,2-di-O-E-feruloyl-β-gentiobiose, 1,2,2'-tri-O-E-sinapoyl-β-gentiobiose, and 1,2'-di-O-E-sinapoyl-2-O-E-feruloyl-β-gentiobiose.

 

Authors: 
S. Moco, J. Vervoort
Authors from the NMC: 
DOI: 
10.1007/978-1-61779-594-7_17
Pages: 
287-316
Book: 
Plant Metabolomics
Publisher: 
Springer
Published in: 
Plant Metabolomics
Date of publication: 
August, 2012
Status of the publication: 
Published/accepted