Tangen ial- low memb ane cla i ica ion o Mal a (Vi is ini e a L.) wine:
incidence on chemical composi ion and senso ial exp ession
Ma in P odano 1*, Ma ga i a Azna 2, Juan M. Cabellos3, Visi ación Vacas3, F ancisco López4,
Ma ía T. He nández5and Ma ía I. Es ella5
1Ins i u o de In es igación en Ciencias de la Alimen ación (CIAL) (CEI, CSIC-UAM), Calle Nicolás Cab e a, 9,
Ciudad Uni e si a ia de Can oblanco, 28049 Mad id, Spain
2Depa amen o de Química Analí ica, Uni e sidad de Za agoza, Campus Rio Eb o, Ma ía de Luna 3,
50018 Za agoza, Spain
3Ins i u o Mad ileño de In es igación y Desa ollo Ru al, Ag a io y Alimen a io (IMIDRA), C a. A-2, km 38.2,
inca “El Encín”, 28800 Alcalá de Hena es (Mad id), Spain
4Depa amen d’Enginye ia Química, Facul a d’Enologia, Uni e si a Ro i a i Vi gili, A . Paisos Ca alans, 26,
43007 Ta agona, Spain
5Ins i u o de Ciencias y Tecnología de Alimen os y Nu ición (ICTAN, CSIC), Calle Juan de la Cie a, 3, 28006
Mad id, Spain
*Co esponding au ho ([email p o ec ed])
Mal a whi e wine (Vi is ini e a L.) was cold se led (CSW) and cla i ied by angen ial- low memb ane il a ion
(TFMF). A 500 kDa molecula mass cu -o memb ane was used. Fil a ion lux o 49-48 L/hm2was achie ed a
ansmemb ane p essu e o 0.7 ba . The ea men p oduced a comple ely cla i ied wine wi h u bidi y o 0.11 NTU,
bu also a 10.3 % loss o p o eins, which could be ela ed o he dec ease o some la ou compounds. The CSW and
he memb ane il e ed wines (MFW) we e assessed by means o hei a oma and phenolic composi ion, as well as
hei senso y p ope ies. The esul s showed ha he gene al physicochemical pa ame e s and mos o he analysed
phenolic compounds we e no o sligh ly (up o 7.6 %) a ec ed by he TFMF p ocess. Ne e heless, he ea men
p oduced an impo an loss o some key a oma compounds: up o 43 % o a y acid and alcohol es e s and up o
26 % o highe alcohols. Mos a ec ed we e a oma species wi h highe molecula masses and lowe pola i ies.
Senso y analysis con i med he global dec ease in wine a oma. TFMF ea men p oduced also an inc ease o 52 %
o he wine benzaldehyde con en .
Mal a wine, cla i ica ion, angen ial- low il a ion, ola ile composi ion, phenol composi ion, ch oma og aphy
BSA: bo ine se um albumin, CSW: cold se led wine, ΔNTU: ne u bididy a e hea es , HPLC: high pe o mance
liquid ch oma og aphy, log P: oc anol-wa e pa i ion coe icien , Lp: memb ane hyd aulic pe meabili y, MFW:
memb ane il e ed wine, NTU: nephelome ic u bidi y uni s, OAV: odou ac i i y alue,
Q : eed low, OT: odou h eshold, PS: polysul one, PTM: ansmemb ane p essu e, PVDF: poly inylidene luo ide,
TFMF: angen ial low memb ane il a ion, TPI: o al phenol index
ABSTRACT
KEYWORDS
ABBREVIATIONS
Recei ed: 29 Ap il 2010
y
Accep ed: 13 No embe 2019
y
Published: 21 No embe 2019
DOI:10.20870/oeno-one.2019.53.4.2480
VINE AND WINE
OPEN ACCESS JOURNAL
725
OENO One 2019, 4, 725-739 © 2019 In e na ional Vi icul u e and Enology Socie y - IVES
INTRODUCTION
Cla i ica ion is an essen ial uni ope a ion o
winemaking. I imp o es no only appea ance o
wines, bu also is e y impo an o hei
physicochemical and mic obial s abili y. This
s a emen is especially heigh ened o whi e
wines, as hey ha e o be clea , wi h u bidi y
lowe han 1 NTU (Ribé eau-Gayon e al.,
2006a). Pos - e men a ion cla i ica ion is usually
ca ied ou a a ious s ages, which can include
na u al o o ced se ling, inning and/o
il a ion. As na u al se ling can ex end oo long
in ime, o ced se ling (cold o cen i ugal) is
p e e ed o indus ial p oduc ion. Al e na i ely,
ining wi h ben oni es, p o eins (egg whi es,
gela ine, caseina es, e c.), silica gel o annins
con ibu es also o speed up he se ling
ea men (Ribé eau-Gayon e al., 2006b; Li a e
al., 2015). In mos o he cases, cla i ica ion ends
up wi h a s age o il a ion. The classic
ea men includes a combined il a ion wi h
cellulose-based il e pla es o dia omaceous
ea hs and ine polishing and mic obial
s abiliza ion o he wine wi h 0.45 o 1.2 µm
po e sizes plea ed memb ane il e s (Hidalgo
Togo es, 2002). In all cases, hese il e aids a e
use ul un il hey clog, which means ha hei use
gene a es was es and subsequen ly,
en i onmen al p oblems.
An al e na i e o hese ea men s wi h a
g owing accep ance in he winemaking indus y
is he angen ial- low (TFMF) o c oss- low
memb ane il a ion. In his case, he gene a ed
u bulen wine eed low d ags insoluble
pa icles om he memb ane su ace and e u ns
hem back o he eed essel, a oiding hei
deposi ion on he memb ane su ace and/o
po es. In he p ac ice, some ine pa icles and/o
colloidal ma e ial build up a ine gel laye on he
memb ane su ace (concen a ion pola iza ion)
(Bake , 2004) ha p oduces a subsequen
dec ease o il a ion lux and memb ane ouling
(El Rayess e al., 2011). Ne e heless,
memb anes employed in angen ial- low
il a ion a e use ul du ing long pe iods (mon hs
and yea s) hanks o he eco e y o he
memb ane pe meabili y a e chemical cleaning
(P odano e al., 2013). Nowadays, memb ane
manu ac u e s o e a wide ange o memb ane
ma e ials (o ganic polyme and mine al sin e ed
laye s), designs (spi al wound, hollow ib e o
capilla y, mul ichannel, e c.) and po e sizes,
which allow many op ions o choice o any
conc e e applica ion. Howe e , his di e si y
also p oduces some con usion o he use s
because o he speci ici y o each applica ion and
he di icul ies o p edic hei e ec . In mos
cases, o cla i ica ion o whi e wines, p oduce s
o e hyd ophilic memb ane ma e ials (cellulose
es e s, poly inylidene luo ide (PVDF),
polysul one (PS), polye he sul one, e c.),
compac memb ane designs (hollow ib e o
spi al wound) and memb anes wi h po e sizes
be ween 0.2 and 5 µm (El Rayess e al., 2011).
Fo whi e wine cla i ica ion, some ul a il a ion
memb anes a e also a ailable. In gene al,
memb anes wi h smalle po e sizes (0.1 o 0.22
µm) gi e be e cla i ica ion e ec s ( u bidi ies
< 0.5 NTU) and mo e s able il a ion luxes, as
he sizes o hei po es a e conside ably smalle
han he sizes o he mos abundan pa icles in
wines and g ape-de i ed p oduc s (Se ano e al.,
1992; Ribé eau-Gayon e al., 2006a; P odano e
al., 2013). The p oblem o such ine cla i ica ion
is ha an impo an pa o he colloidal ma e
can be emo ed also (Ulb ich e al., 2009; El
Rayess e al., 2011; Obe hols e e al., 2013),
leading o dec ease o wine la ou in ensi y. I is
no ewo hy o men ion ha mos bibliog aphic
s udies dealing wi h CFMF conside only
senso y o gene al physicochemical e alua ion
o he cla i ica ion (Feuilla e al., 1987; Flo es e
al., 1991; Se ano e al., 1992; Bu on e al.,
2014) o p oblems ela ed o memb ane s abili y:
ouling, oulan s o memb ane ma e ials (Ve nhe
e al., 1999; Ribé eau-Gayon e al., 2006a).
S udies ela ed o he e ec o TFMF on wine
la ou componen concen a ions and he
in ensi y o hei pe cep ion a e sca ce and in he
cases whe e hey we e a ailable, da a abou he
TFMF p ocess we e no a ailable (Se ano e al.
1984, Flo es e al., 1991; Se ano and Pae zold,
1994). Tha is why he main aim o his s udy
was o e alua e he junc ion o p ocess
pa ame e s, a oma and phenolic composi ion and
senso y e alua ion o a young whi e wine be o e
and a e ine cla i ica ion wi h one o he
memb anes wi h igh es po es a ailable on he
ma ke , he 500 kDa molecula mass cu -o
Romicon memb ane.
Ano he objec i e o his s udy was he chemical
cha ac e iza ion o a wine, p oceeding om he
li le known c . Mal a (Vi is ini e a L.) g apes
(VIVC 7254). This is a whi e a ie y, which
p oduces ui y wines (San os e al., 2004). I s
cul i a ion is admi ed only in o he
denomina ions o o igin ‘Vinos de Mad id’ and
‘La Mancha’. Due o he sca ce in o ma ion
Ma in P odano e al.
© 2019 In e na ional Vi icul u e and Enology Socie y - IVES OENO One 2019, 4, 725-739
726
abou his a ie y in he li e a u e, i s
winemaking quali ies and isual simila i ies o
Ai én (VIVC 157), he mos wildly g own
a ie y o he same egion, i was almos
unknown be o e he las wo decades (Hue a e
al., 1998; San os e al., 2004). Now, his a ie y
is gene ically cha ac e ised and ecognized as
au och hone (Ibáñez e al., 2003; Cabello e al.,
2012). In his sense, he p esen s udy is he i s
mos consis en e o o chemical
cha ac e isa ion o an au hen ic Mal a wine.
MATERIALS AND METHODS
1. G apes
An amoun o 1424 kg o echnologically ipe
g apes (22.3 % o o al soluble subs ances) om
Vi is ini e a L., c . Mal a was ha es ed a he
expe imen al ineya d o he Ins i u o Mad ileño
de In es igación y Desa ollo Ru al, Ag a io y
Alimen a io (IMIDRA), a m ‘El Encín’, Alcalá
de Hena es (Mad id), Spain. G apes we e
handpicked and placed in igid s ackable plas ic
boxes in amoun s o 18 o 20 kg/box o p e en
g ape smashing. A e ha es ing, hey we e
s o ed o e nigh in a cooling chambe a 5 ºC o
main aining hei quali y un il c ushing.
2. Chemicals and eagen s
SO2and analy ical g ade N2 (99.9999 %) we e
pu chased om Ca bu os me álicos (Co nellà de
Llob ega , Spain), a a ic acid wi h 99.5 %
pu i y was om TIDSA (Fuen idueña de Tajo,
Spain) and NaOH and NaClO3we e om
Scha lab (Ba celona, Spain).
Demine alized wa e wi h elec ical conduc i i y
o 7 o 8 µS/cm was ob ained by a e e se
osmosis uni om Tesacua S.L. (Mad id, Spain).
MilliQ-g ade wa e was p oduced in a MilliQ®
In eg al 3 pu i ica ion sys em (Me ck Millipo e,
Bille ica, USA).
Fe michamp ade ma c Saccha omyces
ce e isae yeas s we e acqui ed om Oenob ands
(Mon pellie , F ance).
3. Winemaking
The 1424 kg o g apes we e c ushed and
des emmed in a model NDC18 c ushe (Della
To ola, T e iso, I aly). The ob ained g ape mash
was squeezed in a pilo pneuma ic p ess (Eno
Mundi, Za agoza, Spain) o 3 h and SO2was
added o i o ob ain 80 mg/L o o al SO2. An
amoun o 250 kg o g ape pomace and 1050 L
o g ape mus we e ob ained. The g ape mus
(wi h a densi y (ρ20ºC) o 1.095 g/mL, o al
acidi y o 4.21 g o a a ic acid/L and pH o
3.84) was u he e ige a ed o 4 ºC and kep a
his empe a u e o o e nigh o se ling cou se
solids. Abou 750 L o he clea pa o he mus
we e ans e ed o a 1000 L s ainless s eel essel
and 1.5 g/L o a a ic acid we e added o hem o
inc ease acidi y. Nex , 150 g o Saccha omyces
ce e isae yeas s (Fe michamp om Gis -
b ocades, F ance) we e ac i a ed in 1 L o
dilu ed wi h wa e mus and inocula ed in he
mus o ini ia e e men a ion. Fe men a ion was
ca ied ou a 16-18 ºC un il he o al educing
suga con en dec eased o 1.6 g/L (i.e. 29 days).
Then he clea pa o he wine was ans e ed
( i s acking) o a a iable olume ank wi h
loa ing lid and was le o se le o ano he
26 days. Subsequen ly, he clea pa o he wine
was ans e ed o ano he essel (second
acking) and was le o se le a 4 ºC. Two
addi ional ackings we e done a he 65- h and he
80- h days o winemaking and a e ha , he
cla i ied wine was s o ed a 12 o 16 ºC o
5 mon hs.
4. Tangen ial- low memb ane il a ion
Wine cla i ica ion was ca ied ou by a low
p essu e labo a o y TFMF uni , p o ided wi h a
100 L eed ank, a iable low pe is al ic pump
(Mas e lex I/P, model 7591-07, Cole-Pa me
Ins umen Co., Ve non Hills, Illinois, USA),
inle p essu e gauge (0 o 5 ba ), 1” ex e nal
diame e single ca idge o hollow ib e
ul a il a ion memb anes (RomiconTM, Koch
Memb ane Sys ems, Wilming on, MA, USA),
ou le p essu e gauge (0 o 5 ba ), needle
p essu e con ol al e and a 100 L ecipien
essel. Memb anes we e om hyd ophilic
polysul one, 500 kDa nominal molecula mass
cu o , 0.09 m2o o al il a ion su ace,
1.09 mm inne ib e diame e and 0.45 m o ib e
leng h. Comple ely new memb anes we e used in
his ea men .
Be o e s a ing il a ion, he memb ane ca idge
was condi ioned by consecu i e washings wi h
demine alised wa e a 20, 50 and 20 ºC. An
amoun o 100 L o he cold se led Mal a wine
(CSW) was submi ed o ine cla i ica ion by he
al eady desc ibed TFMF uni . Fil a ion was
ca ied ou by he ba ch concen a ion mode,
ecycling he e en a e in he eed ank and
collec ing he il a e in o sepa a e essel. The
eed low (Q ) was se a 660 L/h (linea eloci y
© 2019 In e na ional Vi icul u e and Enology Socie y - IVESOENO One 2019, 4, 725-739 727
o 2.9 m/s), ansmemb ane p essu e (PTM) o
0.7 ba and empe a u e o 14 o 20 ºC (no
ex e nal e ige a ion). Fil a ion was s opped
when 55 L o cla i ied wine we e p ocessed, ha
co esponds o 17 h o il a ion.
5. Wine sampling
The cla i ied (memb ane il e ed) wine (MFW)
and 10 L o he un il e ed CSW we e bo led wo
days a e il a ion and s o ed a he IMIDRA’s
wine cella a 11-15 ºC un il hei ins umen al
and senso ial analyses. All analyses we e
comple ed in he nex h ee mon hs.
6. Memb ane chemical cleaning
A e il a ion, memb anes we e washed
subsequen ly wi h wa e a 20 ºC o 30 min, and
a 50 ºC o 30 min. Regene a ion was ca ied
ou wi h NaOH solu ion (pH 11) a 50 ºC o 1 h,
NaOH solu ion (pH 11) + 0.1 % sodium
hypochlo i e (NaClO) a 50 ºC o 30 min.
NaOH was emo ed buy displacemen wi h
wa e a 20 ºC un il pH became neu al.
7. Memb ane hyd aulic pe meabili y
de e mina ion
Memb ane hyd aulic pe meabili y (Lp) was
de e mined be o e (L0p) and du ing il a ion
(LFp), as well as a e chemical cleaning (LCp) by
plo ing he co esponding wa e (wine) lux
alues, measu ed a Q o 660 L/h and 20 ºC,
e sus he applied TMP, acco ding o Cassano
e al. (2011).
8. Physicochemical analysis
Hyd ogen ion ac i i y (pH) was measu ed
di ec ly by a FE20/EL20 pH-me e (Me le
Toledo, Spain). To al soluble subs ances (TSS)
we e de e mined by di ec measu emen o wine
samples by hand-held e ac ome e A ago
(model HSR500, Japan) in he in e al o 0 o
32 % (°B ix). Tu bidi y (haze) was de e mined
by a model D-112 u bidime e in he in e al o
0 o 800 NTU (Dinko Ins umen s, Ba celona,
Spain). Ti a able acidi y was de e mined
acco ding o he me hod o he In e na ional
O ganisa ion o Vine and Wine (OIV) (OIV,
2015). To al SO2was de e mined iodome ically
as a sum o he ee and combined SO2,
acco ding o OIV (OIV, 2009a). To al educing
suga s we e de e mined acco ding o he me hod
o OIV (OIVb). Alcohol concen a ion was
measu ed by ae ome y, acco ding o he me hod
o OIV (OIV, 2009c). To al phenρol index (TPI)
o he cla i ied wine was de e mined di ec ly by
measu emen o he abso p ion o dilu ed (1/100)
wi h wa e samples a 280 nm. TPI o non-
cla i ied wine samples we e measu ed a e
il a ion o he dilu ed (1/100) samples h ough
a PVDF memb ane il e wi h a 0.45 µm po e
size. Oc anol-wa e pa i ion coe icien (log P)
was calcula ed by he so wa e MOE (Chemical
Compu ing G oup ULC). To al p o ein con en
was assessed by B ad o d’s me hod using
Coomassie b illian blue eagen as desc ibed in
Salaza e al. (2007) and was exp essed as mg/L
o bo ine se um albumin (BSA) (Bio-Rad
Labo a o ies Inc, Richmond, CA, USA).
P o ein s abili y was ca ied ou by hea es o
bo h MFW and CSW wines, acco ding o Li a e
al., (2015). In b ie , 10 mL o wine samples we e
il e ed h ough a egene a ed cellulose
memb ane il e wi h a po e size o 0.45 μm
(Wha man, England) and hei u bidi ies we e
measu ed by u bidime y. Samples we e hen
hea ed a 80 °C o 2 h in a wa e ba h. A e
incuba ion, hey we e cooled unde ap wa e ,
placed in ice and le in e ige a o o o e nigh
a 4 °C. Finally, samples we e homogenized by
gen le shaking and hei u bidi y was measu ed
again. Wines we e conside ed o be p o ein
uns able when he di e ence be ween haze
be o e and a e hea ing (ne haze) was g ea e
han 2 NTU.
A oma compounds we e analysed by
Pu ge&T ap-GC-MS ollowing he me hodology
de eloped by Azna and A oyo (2007). The ap
(Voca bTM 3000) con ained di e en adso ben s.
Fi s ly, he samples we e dilu ed wi h milliQ-
g ade wa e 1/20 ( / ) o a oid sa u a ion o he
ap by e hanol and possible p oblems due o
oaming. An aliquo o 5 mL was pu ged wi h
helium a low a e o 40 mL/min du ing 20 min
and deso bed a 250 ºC du ing 10 min. Twen y
one ola ile compounds we e analysed:
10 es e s, 7 alcohols, 2 acids, 1 aldehyde and
1 lac one. Iden i ica ion o he compounds was
pe o med by compa ison o hei LRI (linea
e en ion index) alue and mass spec um wi h
hose o pu e e e ence compounds.
Quan i ica ion was ca ied ou by ex e nal
calib a ion cu es, o each o hese compounds.
Odou h eshold (OT) alues o he iden i ied
a oma compounds we e aken om Gu h, 1997,
López e al., 2002; Azna e al., 2003; Cha ez e
al., 2007 and Escude o e al., 2007. Odou
ac i i y alues (OAV) we e quan i ied, di iding
Ma in P odano e al.
© 2019 In e na ional Vi icul u e and Enology Socie y - IVES OENO One 2019, 4, 725-739
728
he concen a ion o each a oma compound by i s
OT alue.
Indi idual phenolic compounds we e de e mined
by HPLC, acco ding o Fe nández de Simón e
al., 2003.
Losses o each measu ed pa ame e be o e and
a e TFMF we e calcula ed acco ding o he
nex equa ion:
Loss (%) = 100 (CCSW-CMCW)/CCSW,
whe e CCSW is he concen a ion o he measu ed
pa ame e in he cold se led wine be o e TFMF
and CMCW is he concen a ion o he same
pa ame e in he il a e o he memb ane
cla i ied wine (MCW).
Resul s we e epo ed as mean alue ± s anda d
de ia ion (SD) o iplica e analysis o each
sample.
9. Desc ip i e senso y analysis
Desc ip i e senso y analysis was ca ied ou by a
panel wi h en ained judges om IMIDRA.
This panel has been o med since 2003 by
wen y i e as e s a e o y aining sessions
(se en mon hs). Selec ion and o ma ion o
as e s was ca ied ou acco ding o he
in e na ional s anda d ISO 8586:2012. Se en een
la ou desc ip o s we e gene a ed using
38 la ou compounds as desc ibed by A oyo e
al. (2009) and Balboa-Lagune o e al. (2011).
All o hem we e p esen ed o he panellis s a
h eshold concen a ion, as well as wi h
inc easing concen a ions in a con ol syn he ic
wine. Then, as e s we e ained wi h he same
compounds added o eal wines om Ai en
g apes a di e en concen a ions. Today, he
IMIDRA’s senso y panel is o med by nine
pe manen and i e a iable as e s. Tas e
sessions a e ca ied ou usually once o wice pe
week.
Senso y e alua ions we e ealized unde ISO
s anda ds ela ed o me hodology (ISO
3972:2011), senso y analysis ocabula y (ISO
5492:2008) and as ing oom (ISO 8589:2007).
Amoun s o 20 o 30 mL o wine we e se ed in
coded s anda d wine as ing glasses a
empe a u e o a ound 18 ºC. Judges exp essed
hei e alua ion o each ol a i e ( ui y, banana,
alcoholic, o - la ou , oxidised a oma, mic obial
a oma and o e all a oma in ensi y and quali y)
and as e (alcoholic cha ac e , acidi y, ui iness,
ege al/g ass, bi e , body, sal y and global as e
quali y) desc ip o wi hin a scale om 1 (low
in ensi y) o 10 (high in ensi y). Analysis was
ca ied ou a duplica e.
10. S a is ical analysis
All analy ical da a we e subjec ed o analysis o
a iance ANOVA (SPSS, Chicago, Illinois, USA),
using he Duncan’s es o he es ima ion o
signi ican di e ences be ween samples wi h
p obabili y o 99.5 %.
RESULTS AND DISCUSSION
1. Winemaking
In gene al, he main physicochemical pa ame e s
o he s udied Mal a wine accomplish he
es ablished o wines om denomina ion o
o igin ‘Vinos de Mad id’ speci ic cha ac e is ics
(Hue a e al., 1998; Gil e al., 2006; Co de o-
Bueso e al., 2013; Co de o-Bueso e al., 2016):
d y wine wi h mode a e acidi y, ela i ely low
pH and phenolic con en and mode a e o high
ui y la ou (Table 1). Howe e , wha
de e mines he singula i y o his g ape a ie y
om echnological poin o iew was i s high
suscep ibili y o oxida ion. This phenomenon
can be pe cei ed easily, as i has a di ec e ec
on wine colou and a oma: colou changes om
yellow/g eenish o g een/g ey and he
cha ac e is ic ui y odou dec eases, whe eas
some unpleasan oxida i e la ou s a ise. Once
s a ed oxida ion, i is di icul o s op i . Tha is
why much ca e should be aken o a oid i .
Gene ally, se ling and cla i ica ion should be
ca ied ou in he sho es possible ime, a as
possible as low empe a u es, a oiding
manipula ions causing ae a ion and concen a-
ions o ee SO2lowe han 30 mg/L.
2. Tangen ial- low memb ane cla i ica ion
2.1. E ec on he main p ocess pa ame e s
The kine ics o he il a ion lux du ing he
memb ane cla i ica ion p ocess is shown in
Figu e 1.
Fil a ion s a ed a lux o 52 L/hm2and
inished in 17 h a 48 L/hm2. This is a ela i ely
low il a ion a e, which can be a ibu ed
mos ly o he low ope a ing PTM applied in his
ea men . Li e a u e da a (Se ano e al., 1992)
show luxes o 57 o 170 L/hm2(a PTM o 0.7 o
1.3 ba ) du ing cla i ica ion o whi e wines and
e e o conside ably highe ini ial lux declines
o newly used memb anes wi h losses o mo e
© 2019 In e na ional Vi icul u e and Enology Socie y - IVESOENO One 2019, 4, 725-739 729

han 30 L/hm2in 1 h (El Rayess e al., 2011).
The obse ed e y small ini ial lux decline o
he il e ed he e wine can be a ibu ed mos ly o
he low load o suspended solids, esul ing a e
he exhaus i e cold se ling ca ied ou be o e
ine il a ion ( he cold se led wine u bidi y was
24 NTU, Table 1).
Figu e 2 shows ha he hyd aulic pe meabili y
o he s udied memb ane dec eased down o
139 L/hm2ba du ing he il a ion p ocess, wha
means mo e han 75 % loss o pe meabili y.
A e chemical cleaning, hyd aulic pe meabili y
g ew up o 516 L/hm2ba , wha co esponds o a
conside able loss o 20 % o he ini ial wa e
lux. The chemical egene a ion p ocedu e was
epea ed again, bu i did no imp o e he
hyd aulic pe meabili y o he memb ane, which
means ha his loss was i e e sible.
3. Quali y o cla i ica ion
Quali y o cla i ica ion should be unde s ood as
he ex en o emo al o he solid phase om a
suspension and is ela ed di ec ly o he u bidi y
o he il a e. Good cla i ica ion o whi e g ape
mus s and wines is conside ed when hei
u bidi y’s a e lowe han 1 NTU (Ribé eau-
Gayon e al., 2006a). In his sense, he used he e
500 kDa polysul one memb ane p oduced an
excellen cla i ica ion o he ea ed wine, gi ing
a u bidi y o 0.11 NTU (Table 1).
Ne e heless, quali y c i e ia should conside
also a maximum e en ion o he senso y
p ope ies and he genuine chemical composi ion
o he cla i ied wine. In his con ex and wi h
e e ence o he basic physicochemical
pa ame e s o he wine, he 500 kDa memb ane
did no p oduce any signi ican e ec on mos o
he measu ed pa ame e s (Table 1). Only mino
diminishing (up o 5.5 %) o he o al wine
acidi y and o al phenol index (TPI) we e ound,
due mos p obably o he loss o some ola ile
acids, such as ace ic and hexanoic acids (Table 2)
and some hyd oxycinnamic acids ( ans-Fe a ic
and Ca eic), (Table 3), possibly associa ed o he
10.3% loss o he wine p o ein (colloidal)
ac ion.
The ea men p oduced also a signi ican d op o
he hea induced u bidi y (ΔNTU) ( om 2.2 o
0.88 NTU), which is indica i e o some haze
s abiliza ion e ec (ΔNTU<2). Ne e heless, his
inding has no big ele ance o he wine p o ein
s abili y, as he un il e ed wine (CSW) had a
ΔNTU alue e y close o he es ablished limi
(2.2). Indeed, he accumula ed expe ience wi h
Ma in P odano e al.
© 2019 In e na ional Vi icul u e and Enology Socie y - IVES OENO One 2019, 4, 725-739
730
FIGURE 1. Pe mea e lux kine ics o Mal a
wine du ing cla i ica ion by he 500 kDa hollow
ib e memb ane (Q 660 L/h, PTM 0.7 ba ,
empe a u e 14 °C).
TABLE 1. Main physicochemical pa ame e s o he s udied cold se led (CSW) and memb ane il e ed
(MFW) wines.
Compound CSW MFW
pH 3.34 ± 0.02a 3.31 ± 0.02a
To al acidi y (g/L) 5.6 ± 0.2a5.2 ± 0.1b
Reducing suga s (g/L) 1.6 ± 0.4a1.6 ± 0.2a
TPI 5.74 ± 0.01a5.47 ± 0.02b
E hanol (%, / ) 12.84 ± 0.03a12.81 ± 0.04a
Glyce ol (g/L) 7.85 ± 0.02a7.86 ± 0.04a
F ee SO2 (mg/L) 45 ± 3a42 ± 2a
To al p o eins (mg/L o BSA) 9.56 ± 0.15a 8.58 ± 0.12b
Tu bidi y (NTU) 24.0 ± 0.3a 0.11 ± 0.07b
Ne u bididy a e hea es (!NTU) 2.20 ± 0.08a 0.88 ± 0.02b
bDi e en le e s wi hin he same ow indica e signi ican di e ences (p ≤ 0.05).
Mal a winemaking a he IMIDRA’s wine cella
shows ha hese wines ha e high p o ein haze
s abili y and usually do no need ben oni e
cla i ica ion (no published da a).
Mos ele an was he e ec o his ea men on
he senso y p ope ies o he s udied wines.
Figu e 3 shows he incidence o he TFMF
p ocess on he main ol a i e pa ame e s and
Figu e 4, on he main as e pa ame e s.
Visual e alua ion o he un il e ed Mal a wine
showed ha i was eiled o clea wi h
mode a ely in ense yellow/g eenish colou (da a
no shown). The ol ac i e phase e ealed a high
ui y a oma wi h a clea p edominance o
banana, and a li le bi lowe in ensi ies o
pineapple, apples and pea s. Alcohol odou also
was mode a ely app eciable. Nega i e odou
cha ac e is ics, such as oxida ion, chemical and
mic obial o - la ou s we e ound a e y low
le els. The as e phase showed a p edominan
ui y la ou wi h clea exp ession o banana
a oma on he backg ound o pineapple, apple and
pea nuances. Alcohol as e was mode a e and
acidi y, balanced. P obably, he mos impo an
pa icula i y o his wine was he low in ensi y
sal y as e, which appea ed a a la e s age o he
as ing, oge he wi h low in ensi y bi e ness.
These ine sal y and bi e as es a e e y speci ic
o his a ie y and can be used, oge he wi h he
high banana a oma, o i s di e en ia ion. Fou
o he en as e s de ec ed also he p esence o
ace ic acid a oma in he wine.
On he o he hand, he TFMF p ocess p oduced a
no iceable dec ease o he global a oma quali y
and in ensi y (11.4 o 14.5 %, espec i ely),
which was exp essed mainly wi h loss o
ui iness (15.3 %) and especially, wi h he main
ui y componen s: banana (13.2 %), apple and
pineapple a oma (da a no shown in his igu e).
Howe e , he p ocess p oduced also some
posi i e e ec s, educing a small ex en s some
o he nega i e ol a i e pa ame e s, mainly hose
ela ed o oxida ion (Figu e 3), ege able/
he baceous (Figu e 4) and ace ic acid odou s
(Table 2), bu hey we e a e y low de ec ion
le els. Wi h espec o he es o he s udied
as e pa ame e s, he il a ion p ocess dec eased
he global quali y and ui iness o he wine, bu
his e ec was no so clea ly de ined by judges,
as hose co esponding o he a oma. In gene al,
he 500 kDa polysul one memb ane p oduced an
© 2019 In e na ional Vi icul u e and Enology Socie y - IVESOENO One 2019, 4, 725-739 731
FIGURE 2. Hyd aulic pe meabili y o he
500 kDa hollow ib e memb ane be o e (L0P) and
du ing il a ion (LFP) and a e chemical cleaning
(LCP) (Q 660 L/h, empe a u e 20 ºC). FIGURE 3. Main ol a i e pa ame e s o he
s udied cold se led (CSW) and memb ane
il e ed (MFW) wines.
' ' ' ' ' ' ' ' ' ' ' '
' ' ' '
' ' ' ' ' ' ' ' ' ' ' '
' ' ' '
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
FIGURE 4. Main as e pa ame e s o he s udied
cold se led (CSW) and memb ane il e ed (MFW)
wines.
' ' ' ' ' ' ' ' ' ' ' '
' ' ' '
' ' ' ' ' ' ' ' ' ' ' '
' ' ' '
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
'' ' ' ' '
''''
'' ' ' '' ' ' ' ' ' '
''''
Ma in P odano e al.
© 2019 In e na ional Vi icul u e and Enology Socie y - IVES OENO One 2019, 4, 725-739
732
CSW MCW ACloss OT OAVCSW OAVMCW OAVloss Log P (o/w) MM
(mg/L) (mg/L) (%) (mg/L) (-) (-) (%) (-) (Da)
E hyl bu y a e ui y, apple 0.334 ± 0.011 0.298 ± 0.016 11 0.020 17 15 11 1.44 116
E hyl iso ale a e ui y, banana 0.003 ± 0.001 0.002 ± 0.014 16 0.003 0.92 0.77 16 1.80 130
Isoamyl ace a e banana, swee , ui y 6.058 ± 0.511 4.060 ± 0.323 33 0.030 202 135 33 1.94 130
E hyl hexanoa e ui y, pineapple 0.465 ± 0.045 0.302 ± 0.038 35 0.005 93 60 35 2.33 144
Hexyl ace a e swee , ui y 0.176 ± 0.005 0.108 ± 0.007 39 0.002 88 54 39 2.47 144
E hyl oc anoa e ui y, banana, pea 0.402 ± 0.066 0.228 ± 0.056 43 2 201 114 43 3.21 172
E hyl decanoa e g ape 0.095 ± 0.034 0.060 ± 0.029 37 0.200 0.48 0.30 37 3.90 200
E hylphenyl ace a e lo al, lilac 0.403 ± 0.012 0.237 ± 0.015 41 0.250 1.6 0.95 41 2.27 164
Die hyl succina e ui y, melon 2.207 ± 0.460 1.740 ± 0.389 21 1250 0.002 0.001 21 1.26 174
E hyl lac a e ui y, bu e y 15.2 ± 0.4 12.8 ± 0.3 9.0 150 0.1 0.09 16 0.17 118
S es e s 25.3 19.8 22
1-P opanol soap, a y 61.1 ± 1.8a63.6 ± 2.1a-4.1 314 0.19 0.20 -4.1 0.33 60
2-Me hyl p opanol soap, a y 14.3 ± 1.2a14.6 ± 0.1a-2.1 40 0.36 0.37 -2.1 0.91 74
1-Bu anol soap, a y 0.766 ± 0.031a0.752 ± 0.027a1.8 160 0.005 0.005 1.8 0.84 74
Isoamyl alcohol ancid, usel cheese 147 ± 4a146 ± 5a0.5 30 4.89 4.87 0.5 1.22 88
1-Hexanol ege al, g ass, bi e 1.53 ± 0.05a1.51 ± 0.04a1.1 8 0.19 0.19 1.0 1.88 102
2-Phenyl e hanol lo al, oses 25.4 ± 0.8 17.3 ± 1.0 32 14 1.81 1.23 32 1.50 122
S highe alcohols 250 244 8.9
Ace ic acid inega 443 ± 11 383 ± 15 13 200 2.2 1.9 13 -0.08 60
Hexanoic acid acid, cheese, a y 4.63 ± 0.32 3.73 ± 0.28 19 0.420 11 8.9 19 1.72 116
S ca boxylic acids 448 387 13
Linalool ci us, lo al, esh 0.004 ± 0.001 0.004 ± 0.001 18 0.015 0.29 0.23 18 2.13 154
!-Bu y olac one swee , ca amel 4.24 ± 0.08a 4.35 ± 0.013a-2.6 20 0.21 0.22 -2.6 -0.64 86
Benzaldehyde bi e , almond 0.113 ± 0.019 0.171 ± 0.025 -52 3000 0.04 0.06 -52 1.48 106
Compound
A oma desc ip ion
TABLE 2. A oma composi ion o Mal a wine be o e (cold se led wine, CSW) and a e memb ane il a ion (memb ane cla i ied wine, MCW),
assessed by CG-MS, a oma compound loss (ACloss) a e il a ion, odou h eshold (OT), odou ac i i y alues o CSW and MCW (OAVCSW and
OAVMCW), OAV loss (OAVloss) a e il a ion, oc anol-wa e pa i ion coe icien o he s udied molecules (logP (o/w)) and hei molecula masses
(MM).
aThe same supe sc ip in he same ow means ha he e is no signi ican di e ence be ween alues (p ≤ 0.05).
app eciable educ ion o he ui y la ou o he
s udied Mal a wine.
4. Incidence on a oma composi ion
A oma compounds ha e key con ibu ion o he
senso y p ope ies o wines. Twen y one ola ile
compounds we e iden i ied and quan i ied in
bo h, he cold se led (CSW) and memb ane
cla i ied (MCW) Mal a wines by GC-MS
analysis (Table 2). Among hem, mos di e se
and impo an o he a oma exp ession o he
s udied wines was he g oup o he a y acid and
alcohol es e s wi h a o al amoun o 25.3 mg/L.
The calcula ed odou ac i i y alues (OAV)
showed ha Isoamyl ace a e and E hyl oc anoa e
had he main impo ance o he global a oma,
con ibu ing wi h swee , ui y, banana and pea
a omas. These esul s ag ee wi h he a oma ic
no es ound du ing he senso y analysis. The
E hyl hexanoa e, Hexyl ace a e and E hyl
bu y a e had an impo an addi ional con ibu ion
o he main a oma bouque wi h esh apple,
pineapple, s awbe y and he baceous
conno a ions. The e hylphenyl ace a e had a
ela i ely low concen a ion, bu enough o gi e
some lo al no es o he whole a oma. An
impo an numbe o highe alcohols we e also
de e mined. They we e ound a highe amoun s
(250 mg/L), bu had lowe con ibu ion o he
global wine a oma because o hei lowe OAVs.
Among hem, only he Isoamyl alcohol and
2-Phenyle hanol had ol a i e signi icance, gi ing
usel and lo al conno a ions, espec i ely. O he
quan i a i ely impo an componen s, bu wi h
lowe odou signi icance we e he Hexanoic and
Ace ic acids. The i s one is known o impa
unwan ed a y a oma o wines. I s occu ence
gene ally is ela ed o he pa ial hyd olysis o
E hyl hexanoa e and a highe alues is
indica i e o he ini ia ion o p ocesses o
oxida ion. The low alues de e mined he e o
Hexanoic acid alida e he good s a e o he wine
and mean ha i has no con ibu ion o he wine
a oma. Ace ic acid was majo ola ile compound
wi h an amoun o 443 mg/L. E en i p esence is
usually associa ed wi h ace ic acid bac e ia
ac i i y, amoun s lowe han 500 mg/L has s ill
pleasan odou con ibu ion o he wine a oma.
Expe judges e use wines wi h con en s o
ace ic acid highe han 600 mg/L (Ede , 2006).
Wi h espec o he memb ane cla i ied wine
(MCW), i can be seen ha he TFMF p oduced
an impo an loss o mos o he de e mined
a oma compounds. Mos a ec ed was he g oup
o a y acid and alcohol es e s, which ha e he
highes con ibu ion o he global wine a oma.
Among hem E hyl oc anoa e, E hylphenyl
ace a e, Hexyl ace a e, E hyl decanoa e, E hyl
hexanoa e, Isoamyl ace a e, Die hyl succina e
and E hyl iso ale a e dec eased wi hin he
in e al o 43 o 16 %. The a oma ic alcohol
2-Phenyle hanol and he e pene Linalool should
be included also in o his g oup because o hei
conside able loss o 32 and 18%, espec i ely
and because o hei posi i e aid o wine a oma.
The loss o hese compounds can be di ec ly
ela ed o he dec ease o he pe cep ion o
ui iness and a he banana, apple, pea and
pineapple conno a ion o he ea ed wine and
con i med quan i a i ely he esul s al eady
ob ained om he senso y analysis. Con a y o
his gene al e ec , he loss o 13 and 19 %,
espec i ely, o ace ic and hexanoic acids could
be conside ed i no posi i e, a leas , no
nega i e, as bo h compounds con ibu e o he
wine unwan ed odou s. Ne e heless, i should
be men ioned also, ha a conside able dec ease
o his acids could be impo an o he global
a oma equilib ium, because hey ac also, as a
coun e pa in he hyd olysis o he
co esponding o hem es e s (Be and, 1981;
Edwa ds e al.. 1990). Howe e , i can also be
seen ha he il a ion p ocess did no a ec
mos o he highe alipha ic alcohols, inc easing
hus hei con ibu ion wi h soap, a y, ancid
and usel o e he weakened ui y a omas o he
memb ane cla i ied wine. I is in e es ing o no e
ha hese compounds ha e he smalles
molecula masses (up o 102 Da, Table 2), which
explain qui e easonably he ob ained esul s.
5. Incidence on phenol composi ion
Phenolic compounds ha e impo an
con ibu ion o he senso y (acid and bi e as e),
mou h eel (as ingency) and unc ional ( edox)
p ope ies o wines. Fou een phenolic
compounds we e iden i ied and quan i ied in he
s udied he e Mal a wines by e e sed phase
HPLC analysis (Table 3).
As i can be seen om Table 3, he o al amoun
o indi idual phenolic compounds in he s udied
Mal a wine was ela i ely low: 46.5 mg/L
(20.7 mg/L (Table 3) + 25.8 mg/L o 2-Phenyl
e hanol and E hylphenyl ace a e, Table 2). This
is mos p obably due o he p ope a ie y and/o
o he winemaking p ocess ca ied ou in his
ial, in which only ee unning g ape juice was
used o e men a ion. Mos di e s was he
© 2019 In e na ional Vi icul u e and Enology Socie y - IVESOENO One 2019, 4, 725-739 733