CHANGES IN THE CHEMICAL COMPOSITION OF
VITIS VINIFERA DURING THE RIPENING PROCESS.
Michal Kříha
Supervisor: Ing. David Šebela
THIS PROJECT HAS RECEIVED FINANCIAL SUPPORT FROM THE
EUROPEAN SOCIAL FUND AND FROM GOVERNMENT OF THE CZECH
REPUBLIC
1. INTRODUCTION
Changes in the chemical composition of VItis Venifera in time
1. INTRODUCTION
PHENOLIC COMPOUNDS IN VITIS VENIFERA
catechin
caftaric acid
malvidine
resveratrol
quercetin
2. AIMS


Can the phenolic compounds be detected in vivo?
Is there a difference between single grapes in the
whole cluster?
3. MATERIALS AND METHODS
3. 1. NONINVASIVE METHODS
Upper side
Middle side
Lower side
3. MATERIALS AND METHODS
3. 2. NONINVASIVE METHODS
Spectrometer SM 9000 (psi.cz)
Reflectance
Anthocyanin indices
 1
1 

ARI  R800  

R
R
700 
 550
i  699
Re d

Green
 Ri
i  600
i 599
 Ri
i 500
(Gitelson and Merzlyak, 2001)
3. MATERIALS AND METHODS
3. 2. NONINVASIVE METHODS

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
3. MATERIALS AND METHODS
3. 3. CHEMICAL ANALYSIS
Corrected anthocynanin absorbance
AA = A529-(0,288*A650)
m = Mw*c (mg/g)
c - concentration
B - lenght of cuvette
ε - extinction koeff. 30.000 (for
anthocyanins)
Mw of anthocyanins
(484,8g/mole)
Spectrometer Lambda 35 (perkinelmer.com)
(Sims and Gamon, 2002)
4. RESULTS
4. 1. ESTIMATION OF PHENOLIC COMPOUND IN VIVO (REFLECTANCE)
4
y = 1,0665x + 2,5851
R² = 0,5048
3,5
3
ARI
2,5
2
1,5
White Grape
1
Red Grape
y = -0,1851x + 1,173
R² = 0,1044
0,5
Lineární (White Grape)
Lineární (Red Grape)
0
0,000
0,100
0,200
0,300
0,400
0,500
Anth content(mg/g)
0,600
0,700
0,800
Very low anthocyanin content in comercially available varieties.
Eg. Cabernet Sauvignon contains 1300mg/kg of the sample
(Draghici at al., 2011)
4. RESULTS
4. 1. ESTIMATION OF PHENOLIC COMPOUND IN VIVO (FLUORESCENCE)
CAFTARIC ACID (Hydroxicynnamic acid)
3000
Average Fl.signal 420-424nm, exc.
320nm,
Average Fl.signal 304-316nm, exc. 280nm,
CATECHIN (Flavan 3-ols)
3000
2500
y = 10660x - 0,3553
R² = 0,2948
2500
y = 8598.4x - 1455.1
R2 = 0.792
2000
2000
1500
1500
1000
1000
White Grape
Red Grape
500
White Grape
500
Red Grape
Linear (all)
0
0.15
Lineární (All)
0
0.20
0.25
0.30
0.35
0.40
OD280nm
(Exc. 280nm, emission 310nm)
0.45
0.50
0,05
0,10
0,15
OD320nm
0,20
(Exc. 320nm, emission 420nm)
0,25
4. RESULTS
pH
Sugar content %
3,8
3,7
pH
Anth. content %
16
0,5
14
0,45
Sugar Content %
0,4
Sugar content %
pH
3,5
3,4
3,3
Anth. Content (mg/l)
12
3,6
10
8
6
4
Anth. Content
0,35
0,3
0,25
0,2
0,15
0,1
3,2
2
0,05
0
3,1
Bup
Bmiddle
Bdown
Cup
Cmiddle
Cdown
0
Bup
Bmiddle
Bdown
The biggest sugar content was
detected in upper sides of the
clusters. They were in the most
advanced state of the ripening
process.
Cup
Cmiddle
Cdown
Bup
Bmiddle
Bdown
Cup
Cmiddle
Cdown
The biggest content of anth.
was measured in red wine.
5. CONCLUSIONS
+ Anthocyanin content can be detected in vivo using reflectance parameters.
- Limited by the anthocyanin concentration.
+ Phenolics can be detected in vivo using:
(exc. 280nm/em.310nm) Catechin
(exc. 320nm/em.420nm) Caftaric acid
- Not limited by phenolic compounds 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.
ACKNOWLEDGEMENTS


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.
REFERENCES
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

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