EFFECT OF RIPENNING ON PHENOLIC

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COMPOUND OF VITIS VINIFERA L.
logo/name
Assoc Prof dr AntoanelaSEED
PopescuGRAPES FROM MURFATLAR
Assist Prof dr Nicoleta Matei
*
VINEYARD
DETERMINED BY A
Assoc Prof dr Georgeta Pavalache
HPLC/DAD
Faculty of Pharmacy, OvidiusSIMPLE
University, Constanta,
RomaniaMETHOD
1

Faculty of Applied Sciences and Engineering, Ovidius University,

Constanta,
Romania
Introduction
2

Results

Identification
and
quantitative
determination
of
the
active
principles of the test solution was
performed
by
comparing
chromatograms
of
standard
mixture (Fig. 1, 2) and the solution
analyzed
(Figure
3-5).
The
reproducibility of the method was
determined by the square Figure
of 3. HPLC chromatogram of first sample (Pinot Noir 27.08.2012)
correlation coefficients (Figure 6).

14.457

15.793

Concerning the total wine growing area,

Romania ranges among the first group of
ten countries of the world. To date, there
has been published research on the
chemical quality of wines produced in
Romania, but there has been no studies
about the phenolic content of grapes seeds
from our knowledge.
This
work
aimed
at
determining
polyphenolic compounds in grape seeds
during ripening of three different varieties
by a HPLC/DAD method. Grape seeds from
three different varieties (Pinot Noir, Coarna
neagra,
Cabernet
Sauvignion)
were
collected in August and September 2012,
during three different times of ripening,
from
Murfatlar, Constanta district, Romania.
m
A
U
2250
The seed extracts were obtained with
2000
ethanol
50% after two different extractions:
1750
maceration
7 day at room temperature, 6 h
1500
cisresveratrol
reflux.
1250
transresveratrol
1000
750
500
250
0
in
0 Figure
2.5 1. HPLC
5 chromatogram
7.5
10 of trans-resveratrol
12.5
15 after
17.5 exposure
20 m
to UV

Figure 2 Chromatogram of standard mixture

Figure 4. HPLC chromatogram of second sample (Pinot Noir 10.09.2012)

Figure 5. HPLC chromatogram of 17-th sample (Coarna Neagra
04.09.2012)

The active principles determined were higher in the samples obtained by

maceration of Pinot Noir and Cabernet Sauvignon varieties and smallest of
Coarna Neagra variety. The maceration process was more efficient than
refluxing. This fact was observed by methyl gallic acid and ferullic acid
which are present in maceration samples and are not in refluxing samples.
Ellagic acid is in excess in samples obtained by maceration when gallic acid
is in excess in some samples obtained by refluxing.

Figure 6 The total polyphenol content of seed grapes

Research
A HPLC system (Agilent 1100/1200) with
quaternary pump, DAD, autosampler and
5m Zorbax Eclipse XDB-C18 (250 x
4,6mm, Agilent Technologies) column, was
used for analysis of phenolic compounds.
Also,
total
polyphenol
content
was
determined spectrometrically using FolinCiocalteus method. The absorbance was
measured
at
725
nm
in
UV-VIS
spectrophotometer Jasco V-630.
HPLC analysis of seed extracts showed that
all extracts contain high levels of total
phenolic
compounds.
The
following
phenolic compounds were founded: trans
resveratrol, cis resveratrol, chlorogenic
acid, caffeic acid, cinnamic acid, vanillin,
gallic acid. The retention times were
between 0.9900.025minutes for gallic
acid
and
15.8670.007minutes
for
cinnamic acid. The amount of polyphenols
in grape seeds reaches the value of
8523.66mg ellargic acid/100g product of

Figure 9. The content of two polyphenols in grape

seeds

Figure 7.The content of six polyphenols in grape seeds

The content of polyphenols is higher

in samples obtained by maceration.
Polyphenols
content
increases
during ripening Pinot Noir and
Coarn neagr and maximum midripening for Cabernet Saugvignon
(figures 7 and 8). Total polyphenol
content was between 1.1489
0.0118 and 2.4659 0.0204
mg/100g.

Conclusion
By HPLC method was founded: trans resveratrol, chlorogenic acid, caffeic
acid, gallic acid, ferulic acid, 3 methyl gallic acid, p coumaric acid, ellagic
acid. Both methods of producing a polyphenol concentrates can be
considered good and can result in the production methods of the
standardized extracts of some active principles, and particularly resveratrol.
Polyphenol content varies differently depending on cooking time for
different varieties. Polyphenol content also vary depending on ripening time
and variety.