A rapid and sensitive method for the simultaneous analysis of aliphatic and polar molecules containing free carboxyl groups in plant extracts by LC-MS-MSReport as inadecuate




A rapid and sensitive method for the simultaneous analysis of aliphatic and polar molecules containing free carboxyl groups in plant extracts by LC-MS-MS - Download this document for free, or read online. Document in PDF available to download.

Plant Methods

, 5:17

First Online: 25 November 2009Received: 31 July 2009Accepted: 25 November 2009

Abstract

BackgroundAliphatic molecules containing free carboxyl groups are important intermediates in many metabolic and signalling reactions, however, they accumulate to low levels in tissues and are not efficiently ionized by electrospray ionization ESI compared to more polar substances. Quantification of aliphatic molecules becomes therefore difficult when small amounts of tissue are available for analysis. Traditional methods for analysis of these molecules require purification or enrichment steps, which are onerous when multiple samples need to be analyzed. In contrast to aliphatic molecules, more polar substances containing free carboxyl groups such as some phytohormones are efficiently ionized by ESI and suitable for analysis by LC-MS-MS. Thus, the development of a method with which aliphatic and polar molecules -which their unmodified forms differ dramatically in their efficiencies of ionization by ESI- can be simultaneously detected with similar sensitivities would substantially simplify the analysis of complex biological matrices.

ResultsA simple, rapid, specific and sensitive method for the simultaneous detection and quantification of free aliphatic molecules e.g., free fatty acids FFA and small polar molecules e.g., jasmonic acid JA, salicylic acid SA containing free carboxyl groups by direct derivatization of leaf extracts with Picolinyl reagent followed by LC-MS-MS analysis is presented. The presence of the N atom in the esterified pyridine moiety allowed the efficient ionization of 25 compounds tested irrespective of their chemical structure. The method was validated by comparing the results obtained after analysis of Nicotiana attenuata leaf material with previously described analytical methods.

ConclusionThe method presented was used to detect 16 compounds in leaf extracts of N. attenuata plants. Importantly, the method can be adapted based on the specific analytes of interest with the only consideration that the molecules must contain at least one free carboxyl group.

List of abbreviationsGC-MSgas chromatography-mass spectrometry

ESIelectrospray ionization

MRMmulti reaction monitoring

FFAfree fatty acids

IAAindole-3-acetic acid

IBAindole-3-butyric acid

ICAindole-3-carboxylic acid

ABAabscisic acid

JAjasmonic acid

JA-Ilejasmonyl-isoleucine

OPDA12-oxo-phytodienoic acid

dnOPDAdinor-OPDA

SAsalicylic acid

Traumatic acid2E-dodecendioic acid

Traumatin12-oxo-10E-dodecenoic acid

Traumatol12-hydroxy-9Z-dodecenoic acid

Royal jelly acid10-hydroxy-trans-2-decenoic acid

GAgibberellin.

Electronic supplementary materialThe online version of this article doi:10.1186-1746-4811-5-17 contains supplementary material, which is available to authorized users.

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Author: Mario Kallenbach - Ian T Baldwin - Gustavo Bonaventure

Source: https://link.springer.com/



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