1. Semi-targeted UPLC-MS analysis of 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. Semi-targeted analysis (vs. targeted) 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 replicates needed (pooled samples), lower column lifetime - challenging data processing

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

6. 3D chromatogram in ESI(-) mode

7. Differential analysis of WT vs. pal1 by UPLC-MS

8. Looking for compounds which acount for the most variability

9. Kaempferol- -Rham-Glc Kaempferol- -Rham-Glc-Rham Kaempferol- -Coum-Rham-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