Abstract
A two-step analytical procedure to identify and quantify relative metabolic abundances in plants subjected to external stimuli is reported. Exploratory factor analysis of UV–Vis spectral fingerprints obtained from statistical simplex-centroid design extracts are used to determine optimized solvent mixtures that characterize specific metabolic changes in plants. PDA-UV–vis analysis is carried out on the most promising solvent extracts in which spectral characteristics of specific chromatographic peaks are used for metabolic class identification and their chromatographic peak heights provide measures of relative metabolic abundances. The procedure is applied to changes in pheophytin a, caffeine, theobromine and caffeoyl derivatives abundances in yerba-mate plants aimed at sexual dimorphism differentiation. Pheophytin a abundances in male plants are found to be 60% higher than in females independent of light access conditions and differing harvest periods. Caffeine abundances in shaded plants were about four times higher in males than females while theobromine was notably absent in only male species. Caffeine and theobromine abundances in plants with high light access showed large changes depending on the harvest time, spring or winter. The difference in male and female caffeoyl derivatives abundances are much greater in shaded plants, between 50 and 70% larger than in plants grown in clearings, 10 and 30%.
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