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Participants/Project Abstracts

Summer 2006 Bioanalytical Science REU Participants

Thao Nguyen , Ohio Wesleyan University

METABONOMICS STUDIES OF ARABIDOPSIS THALIANA USING LC-MS AND LC-NMR

Thao P. Nguyen , Christiana E. Merrywell and Cynthia K. Larive

Department of Chemistry, University of California-Riverside, CA 92521 and Ohio Wesleyan University, Delaware, OH, 43015

Metabonomics, the study of relative changes in levels of endogenous metabolites due to a pertubation, was used to study A. thaliana under the effects of sort ing in hibitors ( Sortins ), to understand the metabolic pathways through which these compounds exert their effects. Sortin-1 and Sortin-2 were previously found to cause reversible defects in vacuole biogenesis and root development (Zouhar et al., 2004). A. thaliana seedlings were grown in agar containing various doses of Sortin-1 or Sortin-2 . The statistical analysis of 1 H NMR spectra of whole extracts was performed using Principle Component Analysis (PCA), with the scores plots representing the variance between samples and the loadings plots highlighting the peaks that account most of the variance. To aid in the identification of unknown peaks, a control plant was extracted in 50:50 acetonitrile/water, 0.1% formic acid. This extract was subjected to a reverse-phase HPLC separation and spectra of the metabolites obtained using loop-storage/transfer LC-NMR and LC-MS. These coupled techniques showed significant advantages for the qualitative identification of metabolites over conventional 1 H NMR analysis of whole extracts because metabolites were grouped into classes of similar hydrophobicity and simpler NMR spectra are obtained. Essential amino acids, sugars, and many other key metabolites were among those identified.

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