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Semi-targeted UPLC -MS analysis of phenylpropanoids in Arabidopsis

Semi-targeted UPLC -MS analysis of phenylpropanoids in Arabidopsis. Ji ří Grúz, L G R. Why phenylpropanoids?. they affect growth, development and reproduction (Huang et al., 2010). Why phenylpropanoids?.

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Semi-targeted UPLC -MS analysis of phenylpropanoids in Arabidopsis

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  1. Semi-targeted UPLC-MSanalysisof phenylpropanoids in Arabidopsis Jiří Grúz, LGR

  2. Why phenylpropanoids? • they affect growth, development and reproduction (Huang et al., 2010)

  3. Why phenylpropanoids? • they may be involved in cell proliferation and differentiation (Tamagnone et al., 1998; Tsukagoshi et al., 2011) • they are induced by cytokinins (Deikman et al., 1995)

  4. Pros fast analysis of all detectable metabolites in a simple run effective selection of metabolites which account for the most variability in the dataset most suitable for abundant compounds, such as phenylpropanoids Cons low sensitivity with many instrumentations complicated use of internal standards more technical replicatesneeded (pooled samples), lower column lifetime challenging data processing Semi-targeted analysis (vs. targeted)

  5. CRITICAL! Semi-targeted UPLC analysis Statistical software, multivariate analysis (MVA) Biological replicates Data export Pooled samples Data filtering, chromatogram building & deconvolution peak detection & alignment Extraction Randomiz., UPLC-MS Raw data export

  6. 3D chromatogram in ESI(-) mode

  7. Differential analysisof WT vs. pal1 by UPLC-MS

  8. Looking for compounds which acount for the most variability

  9. Kaempferol- -Coum-Rham-Rham Kaempferol- -Rham-Glc ?? Kaempferol- -Rham-Glc-Rham Looking for compounds which acount for the most variability

  10. What is it useful for? • to compare “all” metabolites in various genotypes or treatments • to find compounds which are affected the most • to identify and quantify interesting candidates

  11. Thank you

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