IMPROVING THE TECHNOLOGIES FOR THE PRODUCTION OF WHITE WINES IN THE

CONDITIONS OF AN INDIVIDUAL HOUSEHOLD IN THE COMMUNE OF COTNARI

 2. Materials and methods
2.1. Grape samples and winemaking.
Feteasca albă grapes were harvested on 30
september 2019, at full maturity from Cotnari
vineyard, that together with Tokay- Hungary,
Rheingau-Germany, Champagne-France
located at the northern limit of economic
cultivation of vines (45-50 º northern latitude).
The grapes were processed in an different
way, not by classic white wine technology; the
difference was that ripe grapes were macerated
for four hour of the solid grape parts with the
must beforehand. The treatments applied to
grapes, must and wine were divided into three
distinct parts.
Pre-fermentative treatments: additions 50
mg/L of sulfur anhydride on grapes in crusher—
destemmer for preventing the oxydation of must
and the dose 2 g/ hL pectolytic enzymes
(Zymoclaire PG®, SODINAL) (enzymes were
diluted with warm water 1:10) for juice
extraction during pressing and limpidity, both
less on traditionally vinified grapes. Five
Saccharomyces yeast at a dose 25 g/hL were
inoculated in five samples of must ( Fermactive
AP®,SODINAL - V1, Fermactive Blanc
Aromatique ®,SODINAL -V2, Zymaflore ® X5,
LAFFORT -V3, Zymaflore ® X16,LAFFORT –
V4, Zymaflore ® Delta,LAFFORT – V5, and no
yeast – V6).
Fermentative treatments: At the beginning
and at the half of fermentation were added
nutrient (V1,V2,V3,V4,V5 - Fermactive Blanc
Aromatique ®,SODINAL) at the dose 30 g/ hL
and glutathione (Fresharom ®,LAFFORT) at the
dose 30 g/hL that limits oxidation phenomena
and to preserve flavors. In the trird part of
fermentation were inoculated β-glucosidase
enzymes (Zymovarietal AromaG ®,SODINAL)
with hydrolytic activities intended to improve
the extraction of fermentation precursors. The
temperature of samples during fermentation
fluctuated between 18 and 25ºC.
The fermentation was stopped after 14 days by
is adding 90 mg/L of liquid solution of sulfur
dioxide of 6% concentration at the same time as
the temperature of the wine was lowered to 10
ºC; less for V6 at which the fermentation
stopped spontaneously on the eleventh day
without any intervention.
Post – fermentative treatsments: All
samples was treated with sodium bentonite
(Bentogran ®,AEB) used as fining agent at dose
20 g/hL.
The samples were protected from light and
heat and kept 21 days without any intervention.
All the doses administered are in correlation
with usage specification of oenological products
and with the legislation in force.
2.2. Physical-chemical parameters were
performed according to the International
Organization of Vine and Wine Compendium
methods of analysis: alcoholic strength, total
sugar, free and total sulfur dioxide, total and
volatile acidity, malic acid, density, total dry
extract, non-reducing extract and pH, after the
decarbonisation operation for each sample.
Therefore the Dujardin Salleron ® 2000
distiller was used to determine the alcoholic
strength. The value of the ethyl alcohol content
of the samples is expressed as a percentage of
volume at the temperature of 20 °C.
The determination of total sugar it was carried
out by the Luff-Shoorl method, by dosing the excess
of cupric ions, the Luff Shoorl method, has as a
principle the reaction between the cupric ion and the
reducing sugars, from which results copper oxide and
aldonic acid.
The determination of free sulfur dioxide was
carried out by the iodometric method and is based on
the direct oxidation of free sulfur dioxide with iodine
The determination of total sulfur dioxide in wine
by iodometric method is based on the principle to
release of sulfur dioxide combined with sugars and
acetaldehyde with hydroxide sodium and sulfuric
acid and then a titration with iodine in the presence of
the starch.
The determination of the total dry extract was
done by calculation according to the Tabarié formula.
To calculate the relative density of the aqueous
solution of the extract, corresponding to the
difference between the relative density of the wine
sample and the relative density of the hydroalcoholic
mixture with the same alcohol concentration as of the
wine sample. Based on the relative density thus
calculated, it is deduced the corresponding total dry
extract content from the calculation tables.
The pH was determined with the WTW Ino
Lab® 740 pH meter, which was previously calibrated
with standard solutions of pH 4.01 and 7 at 20 ° C.
For determination of the relative density was
carried out by the densimetric method, the samples
having 20 ºC.
The total acidity was determined by the
titrimetric method which consists of titration
(neutralization) of the wine sample with a NaOH
solution of normal and known factor in the presence
of phenolphthalein as an indicator, after prior
elimination of CO₂.
The analysis of volatile acidity was carried
out by titrimetric method by titration of volatile
acids separated from wine by entrainment with
120 100
100
78
80
Liters
60
40
20
0
Samples with enzymatic treatsment
Misture
3. Results and discussion
in an acidic medium.
water vapor and rectification of vapors,
expressed as a percentage volume at the
temperature of 20°C.
The analysis of malic acid parameter in the
samples it was was carried out using a high
performance liquid chromatography technique. In this
sense a liquid chromatograph Shimadzu series
Proeminence® LC20 was used.
For determination of some esters from samples
a HS20-GC-MS was used. The system uses as carrier
gas the nitrogen and the adsorption material for the
stationary phase is the type of resin 2, 6-diphenylene
oxide. For the concentrating process of the samples
was used a method that takes volatile compounds
from the vapor atmosphere and performs the
concentration of the vapors on the stationary phase.
2.3. Reagents and standard solutions
were an analytical grade and were purchased
from Merck, Germany.
2.4 Evaluation of the volumes of must
obtain was made by analyzing the quantity of
liquid obtained by pressing the misture formed
by must and solid grapes parts (pomace). It can
be shown in Figure 1.
Figure 1. The quantity of must resulted
from pressed grapes (L)
100
69
Samples without enzymatic
treatsment
Must
3.1. Concerning adding pectolytic
enzymes at the start of maceration can facilitate
extraction of must and clarification. Pectinase
activities that can efficiently hydrolyse the pectic
substances present in the juice, allowing
sedimentation of the suspended solid particles.
Due to their ability to degrade cell walls,
pectinases are frequently used to improve the
extraction of skin. The use of pectolytic enzymes
may cause an increase in methanol as a result of
the pectin esterase activity (P.Ribereau-Gayon
et.al, 2006). This treatment also improves the
organoleptical characters (notably structure) of
the wine (Canal Llauberès, 1992). When
enzymatic treatments are applied during
maceration, colour remains more stable during
storage (Salinas et al., 2003; Cayla, 2007).
In samples treated with pectolytic enzymes, a
higher quantity of must was observed, more
precisely 78% from the misture compared to
untreated one which had a 69 % must yeld.
3.2. The physical-chemical parameters of
the wine samples are presented in Table 1.
Comparing the wine samples obtain from
Fetească albă were found major difference
concerning amounts of sugar, total acidity ,
volatile acidity, amount of malic acid and a
big difference between sample V1 and
sample V6 traditionall separately vinified.
Concerning the alcohol strengh
parameter, all samples had more then 11%
vol. alcohol, but can be noticed the
difference between sample V1(12,34%
vol.alc.) and sample V6 (11,2% vol.alc.),
both of musts with 210 g/L of sugar before
fermentation process; also at sample V6 can
be observed that for 1 % vol. alc. it took
more than 17 g/L of sugar. So , there is a
premise that in unsulfited must were
Kloeckera yeasts cells (in their metabolism
are needed 21 grams of sugar to produce 1
milliliter alcohol).
Sulfur dioxide is very important in
microbiological stability, taste, color and
limpidity. Only molecular sulfur dioxide has a
Residual sugars parameter presented
values between V1(2,1 g/L) and V4(16,7
g/L), therefore selected yeasts in the
second one had a slower multiplication
and an “unfriendly” medium.
Also for sample V6, are seen 8,2 g/L
sugars, but the fermentation process had
stopped before the fourteenth day that
means the whole fermentation process was
stucked.
The traditional procedure has been to
allow endemic yeast to conduct
fermentation, therefore a high rate of
indigenous yeast multiplication involved a hight
yeasts population that consumed much of
assimilable nitrogen, and towards the end of
fermentation it no longer existed in the must.
Concerning total acidity of samples
parameters fluctuated between 6,35 g/L
(V5) and 7,0 g/L (V6) , expressed in tartaric
acid.
This is an important acid because
contributes to a wine’s taste, stability, color
and in small quantity makes it unstable (high
pH causes high bacterial activity and wine
degradation). Higher parameter of V6 (0,5
g/L) volatile acidity expressed in acetic acid
show a high presence of acetic bacteria
due to lack of sulfur dioxide does not have an
inhibitory effect like in the other samples
(V1,V2,V3,V4,V5). Over time wine could reach
value 1,5 g/L volatile acidity and could no
longer be consumed.
Malic acid contained identified in samples
vary between 0,17 g/L (V3) and 0 g for V6. So
,that indicate a high presence of
Schizosaccharomyces Lindner yeasts cells or a
malolactic bacteria wich gave malolactic
fermentation.
Lower pH levels situated between 3,18
(V6) and 3,28 (V3) , makes wine stable, and do
not alow the growth of microorganism.; pH
values must be less than 3.4 for white wines to
be stable according to the “Treatise of
Oenology” (Valeriu D.Cotea).
protective role and represents up to 2% of free
sulfur dioxide quantity value.
 Small quantities of free SO2 obtained prevent fermentation or activity of
makes all samples instables to the microorganisms. Acording to the literature all
refermentation and significant accumulations of the other samples wines requires 45-50 mg/L
hydrogen sulfide, less V1 sample (33mg/L) , free SO2 to prevent refermentations.
being a dry wine , 25-30 mg/L it is sufficient to

Table 1 : Physical-chemical parameters of obtained samples

Sample Alc. Reducing Free Total Total Volatile Malic Relative T.D.E N.E. pH
(% substance SO2 SO2 acidity acidity ac. density (g/L) (g/L)
vol.) (g/L) (mg/L) (mg/L) (g (g (g/L) (g/L)
tartaric acetic
ac./L) ac.L)
V1 12,34 2,1 33 76 6,5 0,22 0,2 0,9928 19,6 17,5 3,24
V2 11,95 6,8 21 64 6,4 0,21 0.21 0,9939 25 18,2 3,28
V3 11,83 8,9 22 69 6,42 0,23 0.17 0,9973 27,8 18,9 3,26
V4 11,51 16,7 21 75 6,45 0,18 0.19 0,9986 36,2 19,5 3,24
V5 11,65 11,9 20 70 6,35 0,33 0.23 0,9964 31 19,1 3,25
V6 11,2 8,2 6 12 7 0,6 0 0,9929 25,6 17,4 3,18

3.2. Gas-chromatographic analysis

The gas-chromatographic analysis of Fetească
albă samples showed the presence of 9 esters
which are resulting in fermentation process, and
after that and improve organoleptic properties
and body of the studied wines. They could be
seen the predominance of the compounds that
give the wines the vegetal, fruity and floral
aromas. Table 2 shows the parameters of the
esters that appear in the wine samples.

Table 2. Esters identifified in the analyzed wine samples(mval eg/L)

Esters V1 V2 V3 V4 V5 V6
Isoamyl 23,58±0,14 22,13±0,12 14,22±0,17 12,23±0,11 10,13±0,11 8.21±0,12
acetate
Ethyl 7213,15±0,06 8058,42±0,06 5443,24±0,06 6454,13±0,09 5998,23±0,11 4582,12±0.04
octanoate
Ethyl 2214,43±0,09 2021,23±0,08 2111,17±0,22 2276,21±0,18 1983,27±0,57 997,87±0,33
decanoate
Diethyl 59,54±0,19 48,73±0,19 49,76±0,29 50,79±0,45 56,72±0,34 62,14±0,26
succinate
2-Phenethyl 40,16±0,07 47,12±0,23 45,18±0,64 30,33±0,03 41,16±0,44 20,98±53
acetate
Ethyl laurate 90,67±0,02 87,78±0,43 70,38±0,25 101,58±0,33 132,43±0,05 158,23±0,17
Isopropyl 14,23±0,73 13,44±0,12 15,57±0,09 11,54±0,03 12,12±0,07 13,13±0,41
myristate
Ethyl 10,04±0,34 12,34±0,07 15,57±0,56 14,89±0,21 16,01±0,23 7,32±0,15
palmitate
Ethyl oleate 114,34±0,06 99,89±0,22 110,43±0,37 89.98±0,08 188,67±0,54 110,12±0,31

Analyzing Table 2 it can be observed that some
esters such as: isoamylacetate, ethyl octanoate, ethyl
decanoate, diethyl succinate, 2-phenethyl acetate,
ethyl laurate, isopropyl myristate, ethyl palmitate and
ethyl oleate.
Variation in their concentrations during ageing
is different; esters may increase or decrease during
storage due to chemical esterification or hydrolysis
towards chemical equilibria (Simpson, 1979; Ramey
and Ough, 1980; Zoecklein et al., 1999).
Due to their esterase activities, yeasts form
various esters. Isoamyl acetate is the most important
acetate of higher alcohols ester which improve the
wine with banana aroma. In bigger quantity can be
seen in samples V1 and V2 and sample V6 had a
lower quantity.
Ethyl octanoate and ethyl decanoate are
considered as highly positive flavour attributes
of young wine aroma and introduce floral and
fruity flavour notes (Díaz-Maroto et al., 2005).
In samples V1 and V2 can be observed
significant quantities less for both esters in
sample V6 .
Diethyl succinate is an ethyl ester of
succinic acid and this had small parameters
value because because this type of ester is
characteristic of aged wines. In big quantities it
gives the wine notes of wax and dust.
2-Phenethyl acetate is one of esters which
give to the wine floral, sweet and fruity notes.
Sample V6 contain only 20,98 mg/L, less than
the other samples.
Ethyl laureate is a laureate ester formed by
the esterification between ethanol and laureate.
This ester enriches the wines in floral, fruity,
wood and grass aromas. It can be seen in lower
Chart Title
vegetal
texture10 8 mineral
unctuous 6 citric
phenolic 5 ripe fruits
6 2 2 4
bitter 5 2 exotic fruits
4 Minerality could be observed in V6 sample and
sweet 4 0 7 dried fruits
0 may be due to a higher potassium content.
acid
9 2 2 7 green fruits
5
oak tree hay
honey 7 8wild flowers
spicy mud
quantities in V2 sample (87,78 meq/L) and V3
sample (70,38 meq/L) and in opposition, in high
quantities in V6 sample (158,23 meq/L)
Regardind greasy, fatty, oily notes of
studied wines which are characterized by the
presence of isopropyl myristate it seems that
there are not too many variations between
samples parameters. V4 sample includes only
11,54 meq/L.
For V3 and V5 samples they could be
noticed higher quantity parameters of ethyl
palmitate ester which attributes to the wines a
vanilla balsamic nunace.
Ethyl oleate is a fatty acid ester formed by
the condensation of oleic acid (a fatty acid) and
ethanol. This ester is formed during yeasts
autolysis and gives to the V5 sample fatty and
oily notes.
3.3. Sensory characteristics of analyzed wines
are are given especially by the gustatory and
olfactory senses. Aroma profile is given by the
presence of alcohols, esters, acids and tannins in
wines. The use of different yeasts selected
concomitantly with the use of glucosidasic
enzymes highlighted various aromatic profiles
from fruity, floral notes, representing olfactory
descriptors, to mineral notes and even mud or
fatty notes ,representing taste descriptors .
Concerning of flavor preservation,
glutathione treatment had the expected effect
because the descriptors in the samples could be
easily identified.
In figure 2 are given some descriptors and
their variability from sample to sample.
V1,V2, V4 samples showed exotic fruity
characteristic notes.
V1 ,V6 samples showed characteristic floral notes.
.
Figure 2. Comparative organoleptic analysis of
wines.

V1 V2 V3
v4 v5 v6
 In V6 sample less desirable notes could be noticed in the wine like : dried fruits, mud, vegetal and
bitterness.
The wine from V2 sample showed a structural and onctuous character V3 and V5 samples showed
citric notes.

CONCLUSIONS

1. By applying enzymatic treatments at the start of maceration can

facilitate extraction of must, and β-glucosidase enzymes treatments
improves the extraction of fermentation precursors.
2. Treatments with sulfur dioxide reduce the oxidation, inhibits
the endogenous yeasts activity and bacteria degradation, prevents
refermentations , the formation of hydrogen sulfide and preserve wine
quality.
3. Traditionally obtained wines are unstable concerning
microbiological activity and may have organoleptic defects.
4. The use of selected yeasts makes a substantial contribution to
the production of quality white wines by the fact that fermentations
does not stop spontaneously and improves the organoleptic
characteristics of wines.