1. THIS PROJECT HAS RECEIVED FINANCIAL SUPPORT FROM THE EUROPEAN SOCIAL FUND AND FROM GOVERNMENT OF THE CZECH REPUBLIC Michal Kříha Supervisor: Ing. David Šebela

2. Changes in the chemical composition of VItis Venifera in time

3. catechinresveratrol caftaric acid quercetin malvidine

4.  Can the phenolic compounds be detected in vivo?  Is there a difference between single grapes in the whole cluster?

5. Upper side Middle side Lower side

6. Spectrometer SM 9000 (psi.cz) Anthocyanin indices (Gitelson and Merzlyak, 2001) Reflectance

7. Fluorescence spectrophotometer Jobin Yvon (horiba.com)  We used emission spectra in different excitation wavelenght (230nm, 260nm, 280nm, 305nm, 320nm, 355nm, 390nm, 530nm, 630nm) acoording to absorption maxima of phenolic compounds

8. Spectrometer Lambda 35 (perkinelmer.com) AA = A 529 -(0,288*A 650 ) m = Mw*c (mg/g) c - concentration B - lenght of cuvette ε - extinction koeff. 30.000 (for anthocyanins) Mw of anthocyanins (484,8g/mole) (Sims and Gamon, 2002) Corrected anthocynanin absorbance

9. Very low anthocyanin content in comercially available varieties. Eg. Cabernet Sauvignon contains 1300mg/kg of the sample (Draghici at al., 2011)

10. (Exc. 280nm, emission 310nm)(Exc. 320nm, emission 420nm) CATECHIN (Flavan 3-ols)CAFTARIC ACID (Hydroxicynnamic acid)

11. pHSugar content %Anth. content % The biggest content of anth. was measured in red wine. The biggest sugar content was detected in upper sides of the clusters. They were in the most advanced state of the ripening process.

12. + Phenolics can be detected in vivo using: (exc. 280nm/em.310nm) Catechin (exc. 320nm/em.420nm) Caftaric acid - Not limited by phenolic compounds concentration. + Anthocyanin content can be detected in vivo using reflectance parameters. - Limited by the anthocyanin concentration. + Higher sugar content cause was detected in upper sides of the clusters. We suppose it is because the upper side was in more advanced state of maturity. - We have to verify it because it depends on the position of the whole cluster.

13.  I would like to acknowledge to my project leader David Šebela for everything he helped me with.  And I would also like to thank to whole organisation team of Summer schools.

14. Draghici L. et al. (2011) Evolution of polyphenolic compounds during maturity of Cabenet Sauvignon grapes from Dealu Mare vineyard. OU annals of Chemistry 22(1): 15-20 Gitelson, A.A. et al. (2001) Optical Properties and Nondestructive Estimation of Anthocyanin Content in Plant Leaves. Photochemistry and Photobiology, 74(1): 38-45 Lichtenthaler, HK and AR Wellburn (1983) Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochemical Society Transactions 11: 591 - 592. Sims D.A. and Gamon J.A. (2002) Relationships between leaf pigment content and spectral reflectance across a wide range of species, leaf structures and developmental stages. Remote Sensing of Environment 81, 337-354