Accurate LC-ESI-MS/MS quantification of 2'-deoxymugineic acid in soil and root related samples employing porous graphitic carbon as stationary phase and a ¹³C₄-labeled internal standard.
For the first time the phytosiderophore 2'-deoxymugineic acid (DMA) could be accurately quantified by LC-MS/MS in plant and soil related samples. For this purpose a novel chromatographic method employing porous graphitic carbon as stationary phase combined with ESI-MS/MS detection in selected reaction monitoring was developed. Isotope dilution was implemented by using in-house synthesized DMA as external calibrant and ¹³C₄-labeled DMA as internal standard (concentration levels of standards 0.1-80 μM, determination coefficient of linear regression R² > 0.9995). Sample preparation involved acidification of the samples in order to obtain complete dissociation of metal-DMA complexes. Excellent matrix related LOD and LOQ depending on different experimental setups were obtained in the range of 3-34 nM and 11-113 nM, respectively. Standard addition experiments and the implementation of the internal ¹³C₄-DMA standard proved the accuracy of the quantification strategy even in complex matrices such as soil solution. The repeatability of the method, including sample preparation, expressed as short- and long term precision was below 4 and 5% RSD, respectively. Finally, application in the context of plant and soil research to samples from rhizosphere sampling via micro suction cups, from soil solutions and soil adsorption/extraction studies revealed a DMA concentration range from 0.1 to 235 μM.
Department of Chemistry, University of Natural Resources and Life Sciences-BOKU Vienna, Vienna, Austria.
Yvonne
Y
Schindlegger
Eva
E
Oburger
Barbara
B
Gruber
Walter D C
WD
Schenkeveld
Stephan M
SM
Kraemer
Markus
M
Puschenreiter
Gunda
G
Koellensperger
Stephan
S
Hann
D001383:Azetidinecarboxylic Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D002247:Carbon Isotopes / Q000032:analysis* / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods*; D004784:Environmental Monitoring / Q000379:methods; D006108:Graphite / Q000737:chemistry*; D057230:Limit of Detection; D016014:Linear Models; D018517:Plant Roots / Q000737:chemistry* / Q000378:metabolism; D012015:Reference Standards; D015203:Reproducibility of Results; D058441:Rhizosphere; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods; D014908:Triticum
D002247:Carbon Isotopes; D001383:Azetidinecarboxylic Acid; C052864:2'-deoxymugineic acid; D006108:Graphite
13C4-labeled internal standard; 2′-Deoxymugineic acid; Iron acquisition; LC-ESI-MS/MS; Rhizosphere