SURVEY

EFFECTS OF DIFFERENT ENOLOCICAL

TREATMENTS ON DISSOLVED OXYGEN
IN WINES
M. CASTELLARI, B. SIMONATO" G. B. TORNIELLII,
P. SPINELLI1 and R. FERRARINI'*
Università c1i Bologna, Dipartimento cli Scienze clcgli Alimenti, sccic di Cesena,
Via Ravennate 1020, 17023 Ceseua,Italv
D ip a rti m e;i:":.^'il'ff d i v e ron a'
H"lT:,;ì5i'i;i,lì1"
* Corresponcling iruthor: e-ma i 1 : robcrto.ferrra"ità
rini@univr.it

AI]STRACT RIASSUN'I'O

More Lhan I.OOO neaslrrenìents of Sono state ellèttuate piu di I.OOO

dissolrred oxygen in wines were made nrisurazioni di ossigeno clisciolto nel
to evaluate the influence of diffèrent vino per valut.are I'eflètto di diversi
technological practices (filtration. trzrttamenti tecnologici (filtrerzione,
centrifugation, reliigeration, tarrtrnte centrifugazione, refiigerazione, stabi-
stabilizalion, bottling and micro-ox1r- lizzazione tartarica, imbottiglianrent o
qenation) and of diflerent conclitions of e micro-ossigenazione) o cli diverse
nraturation (stainless steel. concrete conclizioni di maturazione (serbatoi
clr wooden barrels) . The micro-oxv- di acciaio, botti di cemento o di legno)
genation of more than lOO red wines sull'ossigenazione dei vini. La micro-
during storage in stainless steel tanks ossigenazione, effettuata in piu di
was ellective in increasing the dissol- lOO campioni di vino rosso. clurante
ved oxygen up to levels comparable to 1o stoccaggio in serbatoi di acciaio,

- Kcv lvorrls: clìssolvecl oxvgerr, cr-rological treatnrurts, oxl.genatior-t, oxvgLìr nlciìsurement, r.r'itrr', r,r'itrcuri'rking -

ttat. J. Food Sci. n. 3. vot. 16 - 2004 387

 those in small wooden barrels. Never-
theless, the storage temperature should
be taken into account to avoid oxygen
accumulation.
sembra essere efflcace nell'aumentare
i livelli di ossigeno disciolto al di sopra
dei valori ossenrati per le botti di legno
di piccolo volume. Tuttavia, anche la
temperatura di stoccaggio dovrebbe
essere tenuta in considerazione al fine
di evitare accumulo di ossigeno.

INTRODUCTION anthocyanins decreases (eaxroR, et aL,

1993). The co-polymerization between
The contact between wine and oxygen anthocyanins and tannins causes a lar$e
maybe considered as a critical point durin$ increase in the degree of condensation
the winemaking process, because oxygen of tannins, inducing a favorable trans-
can signilìcantly influence the wine quality formation from a sensorial point of view.
and its evolution during maturation and Therefore, mild conditions of o\Tgena-
aging. Solubili[. of oxygen fiom air into tion are generally positively regarded for
wine saturated at atmospheric pressure the production of well-aged red wines
is about 6 or 8 rt:rg/L, depending on wine (RIBEREAU-GAYON el al., 1983).
composition and temperature (SINGI.E If wines are stored in wooden barrels,
TON et aI., 1987). Phenolic interactions oxygenation occurs during barrel fìlling
(i.e. their oxidation and polyrnerization) and during opening of filled barrels. This
are initiated and are further promoted by phenomenon is due to a depression cre-
the absorption of oxygen by wine, even if ated in sealed barels during aging by
the rate of progression from monomeric to water evaporation (MouToUNEl' et ol.
polyrrreric pigment fbrms depends largely 1998). When stainless steel tanks are
upon temperature (SOMERS and EVANS, used, moderate or controlled oxygenation
r986). (micro-oxygenation) has been proposed to
Two types of phenolic condensation improve red wine quali[z (PONI'AILLEIì and
normally occur during maturation of red RIBEREAU-GAYON, I 983; MOUTOUNET and
wines. The first is described as a direct MAZAURIC. 2OOl). On the other hand, the
condensation between anthocyanins and absorption of oxygen as a consequence of
(+)-catechin, (-)-epicatechin or tannins various processing treatments (racking,
(BARANOWSKI ANd NAGEI,, I9B3; GUERTìA fìning, pumping, fìltration, treatments
and GLORIES, 1996). The second is the at low temperatures, filling) is gener-
coupled autoxidation of ethanol and ally considered negative for the sensorial
phenolic compounds that produces characteristics of white and rosé wines
acetaldehyde. Acetaldehyde can be in- IMOUTOUNET and MAZAURIC, 2OOl;
volved in the polymerization between RIBEREAU-GAYON et al., 1998).
anthocyanins and other flavonoids or The level ol'dissolved oxTgen and redox
polymeric structures already formed by potential (rH) should be considered as two
direct condensation (SOMERS and EVANS, critical parameters for the control of the
1986: TIMBERI.AKE and BRIDI-E, 1977: winemaking process or for wine treatments
ES,SAFI et al.. 1999). This pollmrerization by micro-oxygenation (VTVAS et ctL., 1992:
can improve color intensity and long- VTVAS and Ct-OzuBS, 1997). Some data has
term color stability (RIBEREAU GAYON been reported about the level of oxygen
eú al., 1983), even if the concentration of that a wine can absorb as a consequence

388 ttal. J. Food Scl. n. 3, vol. 16 - 2004

ol' diflèrent technological treatrnents, but
generally the measurements were car-
ried out on a limited number of samples
(MOUTOUNE"I and MAZAURIC, 2OO I ; VTVAS
et aI., 7997, 1995, 1996; VIVAS and cLO-
RIES, 1993). In a recent study performed
wtth a similar methodologz that was used
ir-r tlre present paper, VIDAL et ctL. (2OOl)
monitorecl the dissolut.ion of-oxJgen in
wine during a large varietv of technologi-
cal steps (i.e. pumping, fìltrertior-r, racking.
centrifugation).
The aim of this stucly r,vas to evalu
ate the influence of single technological
practices on the lerrel ol'dissolved oxygen
in wines. operating under actual win-
ery conditior-rs and on a large number
of wine samples. Various technological
t.reatments (racking, flltration. centrifu-
gation, bottling, et.c.) and rnaturation
conclitions (storage in stainless steel zrnd
concrete tanks or wooden barrels) were
consiclered. Moreover the effect. of nicro-
oxygenation on red wines during storage
in stainless steel tanks was studied in
order to verifv how this technique cotrld
increase the level of dissolved oxvgen.
MATEzuALS AND METFIODS
Wines
This survey was carried out in 68
medium and large-sized Italian wineries
using red ancl white Italian wines ol dil:
fèrent origins (cultivars Merlot, Cabernet
Sauvignon, Sangiovese, Montepulciarno.
Corvinone, Rondinella, Chardonnay, Pi
not,'frebbiano). All wirres were obtained
following the ustral winemaking proc-
esses and contained less than 0.2 ppm
of-dissolved oxygen and less than lOO
rng/L of total sullìtes belbre the begin-
ning of the trials.
Wine treatments
Red (285) and white (245) rvines were
submitted to specilìc enological treal-
ments (Table l)wjth systems, eqtripment
and procedures (to reduce air contact)
commonly used in each winery. Increase
ol-dissolved oxygen (O,,,,.) was calcularted
as the difference between the oxygen
concentration nreasured before (O,,,.,) and
inrmediately a{1er (O,,,) each treat.ment
[O,,'.= O",,.- Ou,,l'
Wine maturation
h-r this case. 56 1 red. well-structured
wines, suitable lbr aging. were consid-
ered. When dissolved oxlrgelt wiìs meas_
r-rred. all rvines had been matured for
4 to 6 rnonths in stainless steel tanks.
concrete tanks or wooden barrels of
large (250 to I,OOO L) and sma.ll (<25O
L) volumes.
Micro-oxyger-ration
Oxygen was supplied continuously with
a Microdue micro-oxygenation systenr
(JU.ClA.S., Verona, Italy) consisting of a
volumel.ric gas-pumping block equippecl
with four micro-porotrs dilfusers. In the
first trial. 118 distinct red wines were
stored in stainless steel tanks at l5'-20"C
and supplied contintrously with different
levels of oxygen (1.2 or 5 mlll-/month).
Dissolvecl oxygerì was then nreasured
after approximately three weeks ol stor-
age. In the second trial, 25 distinct red
wines were sllpplied continuously with
oxygen (3rnL/L/month) during slorage in
stainless steel tanks without temperature
control. The level of dissoh.'ed o\Tgen was
then measured everv month fion-r Seo-
tember to February.
Measurement o[ dissolvecl oxygen
Measurenrent of dissolved oxygen was
carried out using a Micro Logler O' mod.
3650/ I 13 (Orbisphere Laboratories,
Geneva. Switzerland) Calibration was
made belbre each measurement in the
air, taking into account the atmospheric
pressure and t.he wine temperature.
Ital. J. Food Sci. n. 3. vol. 16'2aa4 389
T:rblc I t)es(.ription ol-enological treatlllenls r'"'ith lhe c'orrcspondit]g nr.tmbcr ol rvitre sanlples

Treatmenl Wines Description

Racking (n=26) From tank to tank (bottom-b0ttom),

wine temperature 1 5"-20"C
Racking (n=20) From tank to tank (bottom-uo)
\^/inó lamnèrrirrro 1( -20 ll
Racking (n=1 2) From tank to tank (botlom-bottom),
wine temperalure 5"-1 0"C
/ Pumping (n=B) Different types, wine temperature 15" 20"C
5 "Barrique Batonnage" (n=23) Manual, as slow as possible, 3-5 minutes
6 "Barrique" Assembling (n=5) Mixing wines of different casks after maturation
7 Filtration (n=26) Kieselgur pressure f ilters (various commercial
brands), wine temperature 1 5'-20"C
Filtration (n=1 2) Sheet fillers, plate filters (various commercial brands),
wine temperature 1 5"-20"C
9 Filtration (n=71 ) Membrane cartridges (nominal porosity 1-0.45 mm),
wine temperature 1 5"-20'C
10 Filtration (n=7) Cross{low filtration, spiralate membrane, recycling,
rnrino iomnoratrrro 16 -20'fì

11 Centrif ugation (n=1 a) Vertical (various commercial brands),

r^rina lomnar4l rro 1( -20oll

12 Ref rigeration (n=B) With scraped sur{ace exchanger (f rom 15" to -5"C).
bef ore static tartrate stabilisation
IJ Heat exchange (n=7) From -5' t015"-20"C, after static tartrate stabilisation
14 Continuous (n=7) Contacl System (various commercial brands)
Tartaric Stabilization
tc Electrodialysis (n=1 0) INRA-patent. wine temperature 1 5"-20"C
to Filling (glass bottles) (n=1 36) Different types of f rllers, w;ne temperature 15"-20 C.
Without bottle agitation
17 Filling (glass bottles) (n=1 1 6) As above. with bottle agitation
bef ore oxygen measurement
1B Filling (brick) (n=6) Wine temperature 1 0"-20"C
19 Filling (bag in box) (n=5) Wine temperature 1 0"-20"C
20 Filling (kegs) (n=6) \A/ no fomnoretr rro ( -2Ool-
v r ,, rv rv, |,vv, urul

Cor-rnections between tanks or enologi-
cal equipment and the instmment were
made with PVC pipes; connections and
the measurenrent cell were flushed out
with nitrogen before measurenents.
Dissolved orygen was nìeasured feecling
the nreasurement cell using a peristaltic
pump (Masterflex equipped with tygoltO
tubes) at a flow ol' 150 mllmin until a
stable measllrement was reached. 'lì-tis
method was previouslv tested to verify the
absence ol orygen penneance through the
walls of the tubing. To this purpose" the
dissolved oxygen at dillerent conditions
of cell fèeding was measlrred (Table 2),
and the same oxTgen level was recorded.
These data indicate the etbsence of cross
conlamination througl-r the walls of the
ttrbing. Wines stored in glass bottles,
bricks, bag-in-box and kegs were sanpled
using specifìc der.ices to avoid contact
rvith air and pumped to the neasure-
mer-rt cell as describecl abor.e.
Statistical procedures
Evaluation of lieqtrency distribution.
non-parametric tests (Kruskal-Wallis)

390 ltal. J. Food Scl. n. 3, vol.16 - 2004

 'llrblc 2 trftèct of dilferent nrethods ol-<'ell l'cccling olì oxveen nreastrreruent

Cell feeding Tube length Time of stabilisalion Dissolved oxygen

(m) (mrnl (ppb)

Peristaltic pump 7 40 B
Peristaltic pump 1 3 8
Gravity t4 70 B

and graphical elerboration of clala were esses on the ler.'el ol' dissolved oxyeen
carried out using Sl.artistica 5.0 for Win- in wines were classifìed as -high enrich-
dows (statSoft, Inc., 'l'ulsa, oK. usA). nrent" and "low enricÌrment" treatmenrs
(Figs. 1 and 2).
Racking increasecl the level of oxvgen
RtrSULTS AND DISCUSSION in rvines with a nedian valLre of 370-375
ppb (F-ig. I, #l-2). When wine temperar-
Fronr the end o1- the alcoholic l'er- t.ure was lower t.han lO'C. el,en if care
mental.ion. wine is exposed to manv was exerted in l.ranstèrring the wines
enclìogical operalions that carn increase liom tank to tank (bottom 1o bottont),
the concentration of'dissolved oxygen. the dissolrrecl oxygen increasecl by a 3-
The ell'ects ol' single enological proc- lbld factor, with a nreclian r,'alue ol' l,2gO

F,Tll+!
r:!ùt:tú !::a,j:ri.:a

L: ti *, - I5'i{, p' g 1,1,s 1s{il': i:::'i::ii:a

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Wine Treatments

l-i8. t Ittc'reasc ol'clissoh'ccl ox-vgerì irì wirìes:ìller "tligh Enr-ichrncnl" treiìtlÌìcnts. I Retcking fbottolr
bol1om, l5'20"C);2 Rert'kingflrottoru trp. l5'-2O"C):13 R:rcking[bottonr bo1tonr5" l0-'C):6'ìi]arr-ic1rrc"
zrssetnblittq: ll Centriftrgation: l2 Rclì'igeratioD: lzl Clonlirruous'l'artrìric Stiìbilization: l6-17I.-illino
glarss l-.ottles. (Details rcported in Titl;le l).
I Meclian valrre: I 25'-75" Dcrcerìtilcs.

Ital. J. Food Sci. n. 3, vol. 16 - 2004 391

 tti::i.ì
iii:tli
ffil :l'i*, - i5'ii F,*r':,errii
i;i:i!,1
I hrlerlisrr rralr.lri
trij,:rit

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Wine Treatments
Fig. 2 - llrcreasc of dissoÌr.ecl oxvgcn irì wines:rtter "Lorv Enrichntent" treatntctìls. :l Prtrnping: 5 "Bar-
riqtlc biì t orlrl.tge": 7 I O I'ilt rer ion: 1 3- lleat c-\('hzìrìge: I 5- Electroclialysis; t B- 20 F illing (Dct ails reporte cl
1

in Terble l).
r Medi.ìrì \'iìlucl I 25'-75' per<'enlilcs.

ppb (Fig. l. #3). According to our meas-
urements, the effect of the type of pump
on t.his enrichment can be considered
negligible (Fig. 2, #4). Data reported in
this paper about the dissolulion of oxy-
gen through the studied processes are
in accord with those pre\riously reported
(VIDAL et al.. 2OOl).
The periodic argitat.ion of wine dur-
ing maturation in wooden casks (barrel
"batonnage") procluced an insigniflcanl
increase in dissolved orTgen (Fig. 2" #5).
On the contrary, mlxing wines of dillèr-
ent casks at the end of maturation (Fig. I,
#6) causecl a signilìcant oxlrgen enrich-
ment (median value ol 1,750 ppb).
Filtration is not a critical operation
in comparison wilh wine aeration, since
the nedian increase in oxygen concen-
tration was always less than 3OO ppb
(Fig. 2. #7-lO). Pressure and crossflow
filters girre the highest values, probably
because they required the addit.ion of an
aid-fìlter or partial wine recycling that
can increase Lhe contact between wine
and air. On the contrary the use of centri-
fuges (lìi{. I. H I I ) ntay cituse siqrrificant
rvine oxygen enrichment (median r.alue
of l2OO ppb).
It is commonly recognized that tartaric
stabilization can produce great orygen
absorption; as expecrted, after refiigerat-
ing to -5'C for static stabilization. there
was a meclian increase of clissolved oxy-
gen of 1,265 ppb (Fig. l, #12).
The use ofheat exchangers to recover
cooling power before bottling (Fig. 2,
#13) also produced a slight increatse
in oxvgen in wines (median l5O ppb).
Continuous systems speeded up the
process but in nrany cases produced
complete saturation of wine with
o\ygen (Fig. 1, #14). Furthermore. the
variability of results seems to point
out that many wineries had great dil'-
ficulties in optimizing the procedures
and the equipment to carry out this
process. Only the use o{-electrodialvsis

392 ttat. J. Food Sci. n.3, vol. 16 - 2004

 (Fig. 2, #15) seemed to strongly reduce
oxygen absorption, down to a median
value of 285 ppb.
The different fìlling procedures pro-
duced a variable increase of dissolved
oxygen in wines, depending on the
type of packaging. Filling of glass bot-
tles (Fig. l, # l6) remains a problem in
the winemaking process since a median
enrichment of B3O ppb of o)rygen was
monitored during this step. When the
head space atmosphere was mixed with
the wines before the measurement. the
additional oxygen enrichment was neg-
ligible (Fig. 1 , #17). This indicates that
currently available bottling equipment
was able to reduce the oxygen in the
head space atmosphere, even if other
critical factors should be evaluated in
this process. In particular, the amount
of oxygen dissolved during bottle filling
depends on the technology used (Fig.
3). It can be seen that counterpressure
units caused a considerable increase in
orTgen dissolution. The range of dis-
solved oxygen data is large, suggest-
ing that single operating conditions
can heavily infÌuence the results. A
decrease in dissolved oxygen can be
observed when vacuum counterpres-
sure units or gravity flller coupled with
nitrogen flushing are used.
On the other hand the use of "al-
ternative" containers (Fig. 2, #18-20)
produced lower increases in dissolved
oxygen (median values <100 ppb) in
comparison with those in glass bottles.
This was probably due to the intrinsic.
simplicity of the fìlling equipment usec
with these "alternative" packaging (brick,
bag in box, kegs).
Fig. 4 shows the lrequency distribu-
tion of dissolved oxygen measured in
red wines matured in four categories

5500

4500
--l-
I

c 3500
CJ
i

X I

0) l
2500
o
a
i

l-l-
I
.@

a)
a
o
1 500 It----ll
I

--l--- | I
i'l-11
I

c)

itil"tl
t_tltll
I u
t|---'i I I
500
L_*r___ri l---i o
i

l
ln! Ll
-f Min-Max
t I ll"/o- / A"/o perOenllles
-500
GRAV GRAV_N2 PR PR-VAC n Median value

Fig. 3 - Eflècts of cìiflèrent bottlirìg systems on dissolvcd oxvgen lcvels in wines.

GRAV = greì\'ity filler: GRAV \2 = gr:u.ity fìller rvith Nr: PR = counterpressurc f ìllers: PR VAC = vacuurn/
coLrnterpressure fìllers.

Ital. J. Food Scr. n. 3, vol. 16 - 2004 393

 $J

FB
fi
ilJ

UE
trftoden trreb c35+L
tt- l,t=dian = 35 pÉ
ir

Dissolued o:<ygen {ppbi

Fig. 4 F-requency cìistribulions of dissolved oxlrgen lerrels in recl rvines that 'uvcre slored in titnks or
rvooden barrrels of different volumes.
lStair-rless steel tanks n=175; Concrete tanks n=125; Wooclen barrcl [Volurrc > 25O L) n=88: Wooden
barrcl [Volurle < 25O L) n = 491

of tanks or wooden barrels. The sam- vations and the measurement condi-
ple distribution was not Gaussian, so tions (VIVAS and GLORIBS, 1997).
non-parametric tests (Kruskal-Wallis) Taking into account these data, the
were carried out. These tests revealed micro-oxygenation of wines, stored
significant differences in the level of in stainless steel tanks produced a
dissolved oxygen between all the dif significant effect (p<O.05) on the level
ferent categories of' tanks (p<0.O5). of dissolved oxygen only when 5 mL
Wines stored in wooden barrels of Or/L/nonth were supplied (Fig. 5).
small vollrme had twice as much When operating under these condi-
dissolved oxygen as those matured tions (temperature between I 5"-20"C;
in stainless steel tanks, indicating 5 mL O"/L/month) the median level of
that storage in wood barrels induced dissolved oxygen in wines was compa-
a moderate oxygenation during wine rable to the values measured in wines
maturation. The concentration of stored in small wooden barrels (Fig.
oxygen measured in wines stored in 4) . When the storage temperature was
wooden barrels appears very low in not controlled (Fig. 6), regulation oJ'
comparison with the data reported by micro-oxy$enation was necessary to
VIVAS and GLORIES (1996, 1997), but avoid excessive increases of oxygen
is in agreement with those reported concentration in the liquid phase,
by MOUToUNEI'and MAZAURIC, (2001) which could cause the development
and MOUTOUNET et aL. (1996). This of aerobic microorganisms and/or
can be related to the number of obser- deep wine oxidation.

394 ltal. J. Food Sci. n. 3. vol. 16 - 2OO4

 i:il:l

g
i*i
{}- ELI

q.l

r{
,:4i dfl

EE
"fr
Éi

;l
it'
.i! :r:l
Irri]41

IrF::lii j Ir'=df .t

ì lirl:l*;*r':-:=ltl' ] lri[_"l*,'r1,;,,rtl: f; l-ìL,l*.':]-:*r":fl-r

F-ig. 5 - Vallues of dissolvcd o{rrgen in red wines submittccl to three diffèrent levels of micro-oxygenation
at l5' 20'C. I Median r':rlue; I 25" 75" percentiles.

é:
{i-
r*, 100

t*

'3t
1-J.',
:b-16

"r*.
É!Éo
ài'\
i4
tul

=rÉ

::Lir':a'i!:.!.-.' i l'-l':Iir,l fr,1;,=.,"1s ..-- 1.,*i r..'lr:,-.r..1 ...tlrti:.,!] i ry!ig1;11tr'..

F-ig.6 - Valrres of'dissolved oxlgen in 25 recl wines submittecl 1o :r 3 ni,/L/rttonth micro-oxvgcnation

at thc ,'vinery tenrpcrature. L: Median rraltrc; I 25. 75" percentiles.

Ital. J. Food Sci. n. 3, vot. 16 - 2004 395

 CONCLUSIONS
The results of this sur-vey show that
many enological practices may produce
significant and undesirable increases in
dissolved oxygen levels in wines. Some
technical improvements could surely
ameliorate this situation, but the fact
that there is a wide range of dissolved
oxygen for several enological treatments
su[*{ests that single operating conditions
can heavily influence the results.
Micro-oxygenation may increase the
concentration of dissolved oxygen in
wines maturated in stainless steel tanks
up to levels comparable with those ob-
served in wines matured in small wood-
en barrels. The temperature of storage
should be taken into account to avoid
undesirable oxygen accumulation.
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Guerra C. and Glories Y. 1996. Róle des flavan
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I-onvaud Funel, A., p. 424. Lavoisier Tec&Doc,
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Moutounet M., Ducournau P., Chassin M.
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