Comprehensive LC-MS/MS analysis of nitrogen-related plant metabolites

Author:

Ćavar Zeljković Sanja12ORCID,De Diego Nuria1ORCID,Drašar Lukáš13,Nisler Jaroslav1ORCID,Havlíček Libor3,Spíchal Lukáš1,Tarkowski Petr12ORCID

Affiliation:

1. Czech Advanced Technology and Research Institute, Palacky University , Šlechtitelů 27, 78371 Olomouc , Czech Republic

2. Centre of the Region Haná for Biotechnological and Agricultural Research, Department of Genetic Resources for Vegetables, Medicinal and Special Plants, Crop Research Institute , Šlechtitelů 29, 78371 Olomouc , Czech Republic

3. Isotope Laboratory, Institute of Experimental Botany, The Czech Academy of Sciences , Vídeňská 1083, CZ-14220, Prague , Czech Republic

Abstract

Abstract We have developed and validated a novel LC-MS/MS method for simultaneously analyzing amino acids, biogenic amines, and their acetylated and methylated derivatives in plants. This method involves a one-step extraction of 2–5 mg of lyophilized plant material followed by fractionation of different biogenic amine forms, and exploits an efficient combination of hydrophilic interaction liquid chromatography (HILIC), reversed phase (RP) chromatography with pre-column derivatization, and tandem mass spectrometry (MS). This approach enables high-throughput processing of plant samples, significantly reducing the time needed for analysis and its cost. We also present a new synthetic route for deuterium-labeled polyamines. The LC-MS/MS method was rigorously validated by quantifying levels of nitrogen-related metabolites in seedlings of seven plant species, including Arabidopsis, maize, and barley, all of which are commonly used model organisms in plant science research. Our results revealed substantial variations in the abundance of these metabolites between species, developmental stages, and growth conditions, particularly for the acetylated and methylated derivatives and the various polyamine fractions. However, the biological relevance of these plant metabolites is currently unclear. Overall, this work contributes significantly to plant science by providing a powerful analytical tool and setting the stage for future investigations into the functions of these nitrogen-related metabolites in plants.

Funder

ERDF Programme Johannes Amos Comenius, and by Ciencia de Frontera CONAHCYT-Mexico

Publisher

Oxford University Press (OUP)

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