Profiling polar and semipolar plant metabolites throughout extraction processes using a combined solution-state and high-resolution magic angle spinning NMR approach.
In metabolomic analyses, care should be exercised as to which metabolites are extracted from the sample and which remain in the residue; the remaining metabolites are typically discarded following the extraction process. In this study, nuclear magnetic resonance (NMR)-based metabolomics was used to visualize plant metabolite profiles throughout a series of repeated extraction processes. Metabolites remaining in the extraction residues of (13)C-labeled Arabidopsis thaliana were recovered by repeated extraction using methanol-d(4) and deuterium oxide. The soluble extracts and residual pellets from each step of the extraction process were analyzed by both solution-state and high-resolution magic angle spinning NMR. Metabolic profiling based on chemical shifts in two-dimensional (1)H-(13)C heteronuclear single-quantum coherence spectra allowed the elucidation of both structural and chemical properties. In addition to the metabolite profile, there appears to be a relationship between metabolite structure and behavior throughout the repeated extraction process. These approaches suggest that metabolites are not always extracted in a single step and that the distribution of metabolites in an extraction scenario cannot be predicted solely on the basis of solubility or polarity. The composition of all metabolites in cells influences the solubility of each metabolite; thus, particular attention should be paid because changes in only a portion of the metabolites could influence the entire metabolite profile in a solvent extract.
Sekiyama
Yasuyo
Y
RIKEN Plant Science Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama 235-0045, Japan.
Chikayama
Eisuke
E
Kikuchi
Jun
J
D017360:Arabidopsis / Q000378:metabolism*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*