THIS PROJECT HAS RECEIVED FINANCIAL SUPPORT FROM THE EUROPEAN SOCIAL FUND AND FROM GOVERNMENT OF THE CZECH REPUBLIC

Michal KříhaSupervisor: Ing. David Šebela

Changes in the chemical composition of VItis Venifera in time

catechin

resveratrol

caftaric acidquercetin

malvidine

Can the phenolic compounds be detected in vivo?

Is there a difference between single grapes in the whole cluster?

Upper side

Middle side

Lower side

Spectrometer SM 9000 (psi.cz)

Anthocyanin indices

700550800

11

RRRARI

599

500

699

600Rei

i

i

i

Ri

Ri

Green

d

(Gitelson and Merzlyak, 2001)

Reflectance

Fluorescence spectrophotometerJobin 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

Spectrometer Lambda 35 (perkinelmer.com)

AA = A529-(0,288*A650)

m = Mw*c (mg/g)c - concentrationB - lenght of cuvetteε - extinction koeff. 30.000 (for

anthocyanins)Mw of anthocyanins (484,8g/mole)

(Sims and Gamon, 2002)

Corrected anthocynanin absorbance

y = -0,1851x + 1,173R² = 0,1044

y = 1,0665x + 2,5851R² = 0,5048

0

0,5

1

1,5

2

2,5

3

3,5

4

0,000 0,100 0,200 0,300 0,400 0,500 0,600 0,700 0,800

AR

I

Anth content(mg/g)

White Grape

Red Grape

Lineární (White Grape)

Lineární (Red Grape)

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

y = 8598.4x - 1455.1R2 = 0.792

0

500

1000

1500

2000

2500

3000

0.15 0.20 0.25 0.30 0.35 0.40 0.45 0.50

OD280nm

Ave

rage

Fl.s

igna

l 304

-316

nm, e

xc. 2

80nm

,

White Grape

Red Grape

Linear (all)

y = 10660x - 0,3553R² = 0,2948

0

500

1000

1500

2000

2500

3000

0,05 0,10 0,15 0,20 0,25

Ave

rag

e F

l.sig

nal 4

20-4

24nm

, exc

. 32

0nm

,

OD320nm

White Grape

Red Grape

Lineární (All)

(Exc. 280nm, emission 310nm) (Exc. 320nm, emission 420nm)

CATECHIN (Flavan 3-ols) CAFTARIC ACID (Hydroxicynnamic acid)

3,1

3,2

3,3

3,4

3,5

3,6

3,7

3,8

Bup Bmiddle Bdown Cup Cmiddle Cdown

pH

pH

0

2

4

6

8

10

12

14

16

Bup Bmiddle Bdown Cup Cmiddle CdownS

ug

ar c

on

ten

t %

Sugar Content %

0

0,05

0,1

0,15

0,2

0,25

0,3

0,35

0,4

0,45

0,5

Bup Bmiddle Bdown Cup Cmiddle Cdown

An

th. C

ont

ent (

mg

/l)

Anth. Content

pH Sugar 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.

+ 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.

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.

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