be e ages
Re iew
The mal and Non-The mal Physical Me hods o
Imp o ing Polyphenol Ex ac ion in Red Winemaking
Ma cos Maza 1,2 , Ignacio Ál a ez 2and Ja ie Raso 2,*
1Depa amen o de Ciencias Enológicas y Ag oalimen a ias, Facul ad de Ciencias Ag a ias, Uni e sidad
Nacional de Cuyo, M5528AHB Mendoza, A gen ina
2
Tecnolog
í
a de los Alimen os, Facul ad de Ve e ina ia, Ins i u o Ag oalimen a io de A ag
ó
n-IA2, Uni e sidad
de Za agoza-CITA, c/Miguel Se e , 177, 50013 Za agoza, Spain
*Co espondence: j aso@uniza .es; Tel.: +34-9-7676-2675
Recei ed: 10 May 2019; Accep ed: 18 July 2019; Published: 1 Augus 2019


Abs ac :
Mace a ion- e men a ion is a c i ical s age in he elabo a ion o high-quali y ed wine.
Du ing his s age, he solid pa s o he g ape be ies emain in con ac wi h he e men ing mus
in o de o ex ac polyphenols mainly loca ed in he g ape skin cells. Ex ac ed polyphenols ha e
a conside able impac on senso y p ope ies (colo , la o , as ingency, and bi e ness) and on he
aging beha io o ed wine. In o de o ob ain wines wi h a su icien p opo ion o hose compounds,
long mace a ion imes a e equi ed. The p esence o he solid pa s o he g apes du ing ed wine
e men a ion in ol es se e al p oblems o he wine ies such as p oduc ion capaci y educ ion, highe
ene gy consump ion o con olling he e men a ion empe a u e and labo and ene gy consump ion
o pe iodically pump he g ape mus o e he skin mass. Physical echniques based on hea ing such
as he mo ini ica ion and lash expansion a e cu en ly being applied in wine ies o imp o e he
ex ac ion o polyphenols and o educe mace a ion ime. Howe e , hese echniques p esen a se ies
o p oblems de i ed om he hea ing o he g apes ha a ec wine quali y. A se ies o ecen s udies
ha e demons a ed ha non- he mal inno a i e echnologies such as pulsed elec ic ields (PEF)
and ul asound may ep esen e ec i e al e na i es o hea ing o assis ing polyphenol ex ac ion.
In e ms o gene al p oduc quali y and ene ge ic equi emen s, his e iew compa es hese he mal
and non- he mal physical echnologies ha aim o educe mace a ion ime.
Keywo ds: ed wine; he mo ini ica ion; lash- elease; pulsed elec ic ields; ul asound
1. In oduc ion
Red wine is ob ained om he mus o ed g apes ha unde goes e men a ion oge he wi h he
solid pa s o he g ape be ies. In his s ep, known as mace a ion- e men a ion, suga s o he mus a e
con e ed in o e hanol by yeas , and polyphenolic compounds a e ex ac ed mainly om he g ape
skin and he seeds.
Mace a ion- e men a ion is he mos c i ical s age in he ed winemaking p ocess. I is essen ial o
ob aining high quali y ed wines, bu is also he one ha equi es he mos ene gy and wo k o ce. I is
es ima ed ha abou 64.3% o he o al ene gy needed o p oduce a li e o wine is consumed du ing
he mace a ion- e men a ion s age [
1
]. Polyphenols a e key ac o s in ed wine, since hey a e in ol ed
in i s senso y p ope ies (colo , la o , as ingency, and bi e ness) [
2
], in i s aging beha io , and in
bene icial heal h e ec s a ibu ed o mode a e wine consump ion [
3
]. In adi ional ed winemaking,
in o de o ob ain a inal p oduc wi h high polyphenol con en , he solid pa s o he g ape pomace
emain in con ac wi h he mus du ing he en i e alcoholic e men a ion p ocess (7–10 days), o e en
o e a longe pe iod o ime. Al hough maximum an hocyanin con en and colo in ensi y is al eady
achie ed du ing he i s days o mace a ion [
4
,
5
], he ex ac ion o p ocyanidins and o he la onoids,
Be e ages 2019,5, 47; doi:10.3390/be e ages5030047 www.mdpi.com/jou nal/be e ages
Be e ages 2019,5, 47 2 o 18
which ha e signi ican impac on o he senso y a ibu es such as as ingency and mou h eel, equi es
longe mace a ion pe iods [
6
,
7
]. As hese compounds a e mainly loca ed in he seeds, i s ex ac ion
equi ed he p esence o e hanol o diso ganize he ou e lipidic cu icle su ounding he seeds [
8
].
On he o he hand, in ed winemaking: a oma ic p ecu so s esponsible o he a ie al a omas in
wines a e ex ac ed om he solid pa s o g ape ba ie s, along wi h polyphenolic compounds.
The necessi y o main aining he solid pa s o he g ape be ies in con ac wi h he e men ing
mus leads o se e al issues aced by wine ies in he ed winemaking p ocess [
9
]. I is es ima ed ha
app oxima ely 20% o he e men a ion anks a e occupied by he solid pa s, esul ing in a educ ion
o he e ec i e olume o he anks and, as a consequence, o a wine y’s p oduc ion capaci y. This
issue becomes especially signi ican a he peak o ha es ing, when he e men a ion-mace a ion anks’
p oduc ion capaci y may be exceeded. O he nega i e side e ec s o longe mace a ion pe iods a e
ela ed wi h he di icul y o con olling he empe a u e inc emen as a consequence o he e men ing
ac i i y o he yeas s when he solid pa s a e p esen in he e men a ion anks, as well as wi h he
labo o ce and ene gy consump ion equi ed o pe iodically pump he wine o e he skin mass ha
ises o he op o he e men a ion anks [10].
Di e en s a egiesha ebeenadop ed inwine ies oenhance heex ac ion o phenolic compounds
and o educe he du a ion o he mace a ion- e men a ion s age in ed winemaking [
11
,
12
]. Physical
echnologies based on hea ing, such as he mo ini ica ion and lash expansion, a e cu en ly being
applied in wine ies o his pu pose [
13
]. They p esen a se ies o p oblem such as he di icul y
in ol ed in s abilizing he colo , he loss o a ie al a omas h ough empe a u e inc emen , and he
consump ion o high quan i ies o ene gy [
14
,
15
]. A se ies o s udies ha e ecen ly demons a ed
ha non- he mal inno a i e echnologies such as pulsed elec ic ields and ul asound may ep esen
e ec i e al e na i es o hea ing in he a emp o imp o e polyphenol ex ac ion [
16
–
19
]. This e iew
compa es he mal and non- he mal physical echnologies ha aim o educe mace a ion ime in e ms
o equipmen complexi y, ene ge ic equi emen s, and o e all quali y o he ed wine.
2. The mal Technologies o Imp o ing Polyphenol Ex ac ion
Al hough he hea ing o ed g apes in o de o educe mace a ion has been in es iga ed since he
ea ly 20 h cen u y, he p ocess was no comme cially adop ed un il he 1970s, when indus ial hea ing
sys ems we e de eloped o ha pu pose [20].
In gene al e ms, he p ocess consis s in hea ing g apes o o e 70
◦
C o a pe iod o ime anging
om a ew minu es o se e al hou s. As a consequence o hea ing, he cell en elopes o he g ape skins
a e b aked down, he eby acili a ing he subsequen elease o polyphenols (mainly an hocyanins)
ha a e loca ed inside he cells in o he liquid phase [
21
]. Hea ing also dena u es enzymes such as
polyphenol oxidase, he eby p e en ing b owning. In ac , hea ing was o iginally used o p e en
laccase ac i i y in g apes con amina ed wi h he mold Bo y is cine ea [22].
Al hough gene ally hea ing o he g apes be o e e men a ion is called “ he mo ini ica ion”,
di e en p e- e men a ion hea ing p ocesses a e cu en ly being applied in wine ies. These echniques
can be classi ied in o wo g oups, depending on whe he he cooling o he g apes, simila ly o hea ing,
is conduc ed using hea exchange s, o whe he he cooling is conduc ed in o a acuum chambe .
The i s kind o p ocess is designa ed as “ he mo ini ica ion”, along wi h i s a ia ions, known as
“p e- e men a ion ho mace a ion” (MPC), and “sho - ime-high- empe a u e ea men wi h wa m
mace a ion” (KZHE). The second g oup in ol es he echnique called he mo- lash, lash d
é
en e
o lash- elease.
Be e ages 2019,5, 47 3 o 18
2.1. The mo ini ica ion, MPC, and KZHE
2.1.1. Desc ip ion o he Techniques
The mo ini ica ion, MPC, and KZHE a e p e- e men a i e hea ing echniques; hey all ha e in
common ha he empe a u e o he g ape mash does no inc ease abo e 85
◦
C, and ha hea ing and
cooling a e conduc ed in hea exchange s [23].
In he mo ini ica ion, hea ing up o a ound 70
◦
C is conduc ed o a pe iod o ime o less han
one hou , a e which he g ape mash is p essed o sepa a e he solid pa s and pe o m e men a ion
as o whi e wine. I hea ing a he same empe a u e is ex ended o a longe pe iod o ime (up o
24 h), and he e men a ion is conduc ed in he p esence o absence o he solid phase, he p ocess
is called MPC (“p e- e men a ion ho mace a ion”). A a ia ion o MPC is he p ocess de eloped in
Ge many called KZHE (“sho - ime-high- empe a u e ea men wi h wa m mace a ion”). In he la e ,
e men a ion is conduc ed in he absence o solids a e main aining he g apes a a ound 45
◦
C o
6–10 h a e ha ing hea ed hem o a ound 85 ◦C o 2 min.
2.1.2. Equipmen
The simples and mos inexpensi e hea exchange s used o hea g apes be o e e men a ion a e
ube-in- ube hea exchange s. To p e en blocking p oblems in his hea exchange s, i is equi ed he
applica ion o he ea men o he en i e mix o juice and solid pa s. To sa e ene gy, i is ecommended
o ea he solid pa s a e p e-d aining in o de o minimize he quan i y o ma e ial ha needs o be
hea ed and cooled. In his case, i is ecommended o use a sc aped-su ace hea exchange wi h a
o a ing sha ha imp o es hea ans e o he p oduc . This app oach pe mi s o p ocess he g ape
mash wi h a mode a e deg ee o p e-d aining while a oiding blocking issues.
Di e en app oaches ha e been de eloped o sa e ene gy in he hea ing o he g ape mash by
eco e ing hea . In such sys ems, incoming well-mixed c ushed g apes wi hou any p e-d aining a e
p e-hea ed oge he wi h he c ushed g apes ha ha e al eady been hea ed. In hese sys ems, and in
o de o a oid blocking, spi al hea exchange s o hea exchange s wi h a sec ion o ec angula o
pa allel ec angula channels a e p e e ed.
An al e na i e o he abo e-desc ibed con inuous single pass me hod is o hea he g ape mash
wi h a ube-in- ube hea exchange while eci cula ing hem on a ank. This app oach, gene ally used
in smalle wine ies, esul s in slowe and mo e he e ogeneous hea ing.
Fo ans o ma ion he suga o mus in o e hanol by yeas s du ing e men a ion, empe a u es
be ween 20 and 30
◦
C a e equi ed. The e o e, a e he hea ing pe iod, i is necessa y o cool down he
g ape mass p io o e men a ion. The cooling s ep is conduc ed wi h hea exchange s simila o hose
ha a e used o hea ing.
Fluids used in his ype o equipmen a e ho wa e o s eam o hea ing, and cold wa e o glycol
o cooling.
In gene al, such ins alla ions used o p e- e men a i e hea ing occupy a conside able a ea wi hin
he wine y. The space is equi ed o he hea exchange sys ems as well as o he acili ies designed o
hea and cool he luids.
2.1.3. Impac o he T ea men in he Composi ion o Wine
The main objec i e in using hese p e- e men a ion hea ing echniques is o speed up he ex ac ion
o polyphenols om he g ape skins wi h he pu pose o elimina ing o educing he mace a ion s age.
Howe e , he cha ac e is ics o he inal wine ob ained wi h such hea ed g apes may be a ec ed [
24
,
25
].
As a consequence o hea ing, wild yeas popula ions a e inac i a ed, hus equi ing he addi ion
o mic obial s a e s o igge e men a ion. Gene ally, alcoholic e men a ion is ini ia ed wi hou
p oblems a e p e- e men a ion hea ing. Occasionally a mo e ab up e men a ion han in adi ional
e men a ion is obse ed, p obably ela ed o he elease o nu ien s om he solid pa s o he g apes
as a consequence o hea ing [
26
]. A signi ican inc ease in suga concen a ion, pH, amino acids, and
Be e ages 2019,5, 47 4 o 18
ammonium in he mo ini ied Ca ignan mus was epo ed [
27
]. Bac e ial popula ions o lac ic as well
as ace ic bac e ia a e also inac i a ed, esul ing in wines wi h low ola ile acid con en . To al acidi y
o wine is no usually a ec ed by p e- e men a ion hea ing. Al hough a mo e ele a ed ex ac ion o
ca ions and anions as a consequence o g ape hea ing has been desc ibed, hey p ecipi a e as sal s o
a a ic acid, hus ul ima ely lea ing wines hus ob ained in he same condi ion as un ea ed wines [
28
].
P e- e men a ion hea ing, in which he solid pa s o he g apes a e p essed and e men a ion is
conduc ed in he liquid phase, has he main objec i e o enhancing he ex ac ion o colo om he
skins. The colo inc emen is a consequence o he apid ex ac ion o an hocyanins.
While an hocyanins a e ex ac ed since he i s momen s o e men a ion, la anols equi e he
p esence o e hanol o be ex ac ed.
Picca do and Gonz
á
lez-Ne es [
29
] epo ed ha he ex ac ion o an hocyanins a e
he mo ini ica ion was p ac ically immedia e. As consequence he an hocyanin concen a ion and
he colo in ensi y in he i s days o e men a ion we e 21% and 45% highe , espec i ely, han in
con ol. Mos s udies o he he mo ini ica ion echnique ha e been conduc ed wi h Pino noi due
o he di icul y o ex ac ing an hocyanins om ha g ape a ie y. I has been epo ed ha he
an hocyanin quan i y in he Pino noi a ie y eached a maximum a he onse o e men a ion, wi h a
concen a ion 2 o 3 imes highe han in adi ional e men a ion. A d as ic dec ease in an hocyanins
was obse ed, howe e , owa ds he end o e men a ion [
30
]. S udies conduc ed a labo a o y scale
ha e demons a ed he deg ada ion o an hocyanins due o empe a u e [
27
,
31
]. An hocyanin con en
was a ec ed by he mo ini ica ion when he ea men was e y p olonged, o abo e 70 ◦C.
Conce ning he e ec o p e- e men a ion hea ing on a oma, i has been epo ed ha wines ha e
a s anda dized senso y p o ile o en desc ibed by oenologis s as “banana yogu ” [
32
]. Fo example,
a ie al a oma ic compounds wi h g een peppe a omas (me hoxypy azines) dec eased in Cabe ne
Sau ignon wines when hey we e he mo- ea ed [
33
]. Ge oy e al. [
31
] epo ed ha a hea
ea men a 70
◦
C o wo hou s induced a signi ican loss o se e al g ape-de i ed a oma compounds
( e penols, no isop enoids and some phenols) associa ed wi h an inc ease in
α
- e pineol, guaiacol
and 2,6-dime hoxyphenol, sugges ing he mal deg ada ion. When he mo ini ica ion was applied o
Ca ignan wine a wo di e en empe a u e le els, 50
◦
C and 75
◦
C, and wi hin wo di e en ime
in e als, 30 min and 3 h, he e ec o empe a u e on a oma composi ion was g ea e han ha o
hea ing ime. Wines ob ained om g apes ea ed a 50
◦
C had highe concen a ions o ge aniol,
β-ci onellol, β-damascenone, and 3-me cap ohexanol, in mos cases [27].
Al hough he mo ini ica ion ein o ces an hocyanin ex ac ion, he wines he eby ob ained a e
known o lack colo s abili y and s uc u e. An hocyanins can dec ease due o enzyma ic hyd olysis [
34
],
o combina ion wi h p o eins, o o e- ixa ion wi h solid pa s such as he skin [
35
] and yeas s [
36
].
Since no alcohol is p esen a he ime o hea ing, he wine does no con ain su icien le els o annin
o s abilize uns able an hocyanins and o p o ide s uc u e. As a consequence, wines ob ained by
he mo ini ica ion a e no usually used o aging, bu comme cialized as able wine o e e yday use.
Finally, since annin ex ac ion is much mo e dependen on inc easing e hanol con en o encou age
i s solubiliza ion, one app oach o ob ain a highe ex ac ion o polyphenolic compounds consis s
in e men ing g apes a e hea ing wi h solid pa s o he g apes, as in s anda d ini ica ion wi h
sho e mace a ion ime. This al e na i e was ound o inc ease o al phenolic index, colo in ensi y
and an hocyanins con en in wine 58%, 25% and 45%, espec i ely [29].
2.2. Flash Release
2.2.1. Desc ip ion o he Technique
The p ocess called “ lash elease” o “ lash d
é
en e” consis s in apidly hea ing he g apes a
empe a u es be ween 85–95
◦
C by a di ec injec ion o s eam. G apes a e hen in oduced in o a
acuum ha ins an ly apo izes he wa e , he eby cooling he ea ed g apes and weakening hei
skin cell en elopes by boiling he wa e inside he cells [
37
]. This e ec on he skin cells enhances
Be e ages 2019,5, 47 5 o 18
ex ac abili y in subsequen e men a ion p ocess ha may be conduc ed wi h o wi hou he solid
pa s o he g apes. A modi ica ion o his p ocess is called “hal ” lash d
é
en e [
38
]. I uses a weake
acuum o cool he g ape mash o a ound 50 ◦C ins ead o 30 ◦C.
2.2.2. Equipmen
Flash elease o lash expansion equipmen consis s o a hea exchange and a acuum chambe .
In he hea exchange , he s eam is di ec ly injec ed o he g ape mash. G ape mash is con inuously
mo ed by wo hollow s em auge s h ough which he s eam en e s in o he acuum chambe . Since he
chambe is unde nega i e p essu e (20–25 hPa), he wa e ins an ly e apo a es, while he g ape mash
is simul aneously cooled. The es ima ed amoun o e apo a ed wa e anges be ween 6 o 10% [
39
].
I is condensed in a condense connec ed wi h he acuum chambe , and einco po a ed in o he g ape
mash o ally o pa ially, depending on he amoun o wa e in a gaseous s a e added o he g ape
mash du ing he hea ing p ocess. The lash elease sys em equi es a boile o p oduce wa e apo o
apid hea ing.
2.2.3. Impac o he T ea men in he Composi ion o Wine
I has been epo ed ha he yeas popula ion lag phase be o e s a ing e men a ion is sligh ly
sho e when he g ape mash is ea ed by lash elease, p obably because he ea men has igge ed
he elease o some yeas nu ien s [40].
Cha ac e is ics o wines ob ained by lash elease can be modula ed by conduc ing e men a ion
in liquid phase, o by keeping he solid pa s o he g apes in con ac wi h he liquid phase o di e en
pe iods o ime. I has been obse ed ha lash elease inc eases he ex ac ion o la anols and
la onols om skins a he han om seeds. The e o e, when e men a ion is ca ied ou wi hou
he skins, he concen a ion o annins wi h espec o an hocyanins is low, as in wines ob ained ia
adi ional p e- e men a ion hea ing. The des abiliza ion o g ape skin cell en elopes seems o acili a e
he ex ac ion o annins loca ed in he acuoles o he hypode mal cells o he g ape skins. Howe e ,
he p opo ion o hose annins in he esul ing wine is low compa ed wi h he annins coming om
he seeds, which equi e he p esence o e hanol o be ex ac ed and also a mo e mace a ion ime [
41
].
Mo el-Salmi e al. [
13
] in es iga ed he phenolic ex ac ion kine ics du ing he
mace a ion- e men a ion o G enache mus p e iously ea ed by lash elease. They obse ed
ha he amoun o a ious amilies o phenolic compounds was highe a he beginning o he
e men a ion p ocess in he lash elease ea ed mus han in con ol. On he o he hand, while he
le els o ca echins, la onols, and p oan hocyanidins inc eased du ing e men a ion o lash elease
ea ed mus s, he concen a ion o hyd oxycinnamic acids emained cons an and an hocyanins
dec eased du ing he i s day, and hen hey emained cons an . The inc emen in concen a ion o
galloyla ed uni s inc eased h oughou e men a ion, e lec ing he g adual ex ac ion o seed annins as
he e hanol le el inc eased. The e o e, al hough he e ec o lash- elease on g ape skin cell en elopes
is mo e d as ic han ha o o he p e- e men a ion hea ing echniques, a con ac pe iod o he solid
pa s o he g apes wi h he mus du ing e men a ion a e ea men is equi ed in o de o ob ain
s uc u ed wines wi h la ge amoun s o polyphenols. A he o he ini ica ion p ocess, he wine
ob ained wi h G enache g apes ea ed by lash elease had a o al phenolic index and a colou in ensi y
14% and 9% highe han he con ol wine espec i ely.
The e ec o lash elease on he ex ac ion o a oma ic compounds and a oma p ecu so s has been
also in es iga ed [42]. As compa ed o wines ob ained by o he p e- e men a ion hea ing echniques,
wines ob ained wi h lash elease main ain hei a ie al a oma ic p o ile. The ea men inc eases
he le els o a y acid e hyl es e s and
β
-ionone in G enache wines. On he o he hand, i has been
obse ed ha lash elease may educe he con en o C6 compounds esponsible o he baceous
a omas [43]. This e ec is especially in e es ing when he wines a e elabo a ed wi h g apes ha ha e
no eached hei op imal s age o ma u i y.

Be e ages 2019,5, 47 6 o 18
Wines o di e en a ie ies such as G enache, Ca ignan, Sy ah, and Mou ed e ob ained wi h
lash expansion echnique we e p e e ed o con ol wines in a senso y analysis, especially when he
con ac ime o he solid pa s o he g apes wi h he e men ing mus was ex ended [44].
3. Non-The mal Techniques o Imp o ing Polyphenol Ex ac ion
Non- he mal echnologies ha e been one o he mos equen ly in es iga ed opics in he ield o
ood p ocessing o e he las decades [
45
]. The “non- he mal” concep e e s o a g oup o echnologies
whose e ec s in oods a e simila o hose caused by hea ing, albei a empe a u es lowe han he ones
used in he mal p ocessing. Some o hese ea men s may in ol e hea due o he gene a ion o in e nal
ene gy (e.g., esis i e hea ing du ing PEF). Howe e , hey a e classi ied as non- he mal, because hey
can elimina e o signi ican ly educe he applica ion o high empe a u es in ood p ocessing, he eby
a oiding he dele e ious e ec s o hea on he la o , colo , and nu i i e alue o oods.
The eme gence o non- he mal echnologies can lead o high quali y p oduc s while sa ing ene gy
by imp o ing hea ing e iciency. Mos o hese echnologies a e locally clean p ocesses and he e o e
appea o be mo e en i onmen - iendly, wi h less en i onmen al impac han adi ional ones [
46
].
No el p ocessing echnologies a e inc easingly a ac ing he a en ion o ood p ocesso s, since hey
can p o ide ood p oduc s wi h imp o ed quali y and a educed en i onmen al oo p in , while
educing p ocessing cos s and imp o ing he p oduc s’ added alue.
Due o hei special mechanism o ac ion, pulsed elec ic ields and high-in ensi y ul asound a e
among he non- he mal echnologies ha ha e been mos in es iga ed wi h he pu pose o imp o ing
polyphenol ex ac ion in wine ies.
3.1. Pulsed Elec ic Fields (PEF)
3.1.1. Desc ip ion o he Technique
PEF p ocessing consis s in he in e mi en applica ion o sho du a ion pulses (ms-
µ
s) o high
ol age (kV) o a p oduc loca ed be ween wo elec odes. The applied ex e nal ol age gene a es an
elec ic ield whose s eng h depends no only on ol age in ensi y, bu also on he dis ance be ween
he elec odes. When exposed o a su icien ly s ong elec ic ield, he cell memb ane unde goes a
phenomenon called elec opo a ion, consis ing in he inc emen o cell en elope pe meabili y as a
consequence o he o ma ion o po es in he cy oplasma ic memb ane [47].
I he in ensi y o he elec ic ield is no high enough, o i he exposu e o he elec ic ield
is su icien ly b ie , he memb ane can spon aneously e u n o i s ini ial s a e and emains iable
( e e sible elec opo a ion). Howe e , in ense elec ic ields o longe exposu es can cause i e e sible
elec opo a ion [
48
]. Re e sible elec opo a ion is a p ocedu e ha is ypically used in molecula biology
and in clinical bio echnological applica ions o gain access o he cy oplasm o he in oduc ion o
deli e y
in i o
o d ugs, oligonucleo ides, an ibodies, plasmids, e c. Howe e , he main applica ions
o PEF in he ood indus y aim o cause i e e sible elec opo a ion o he cell memb anes. I has
been demons a ed ha i e e sible modi ica ion o he pe meabili y o cell memb anes can inac i a e
ege a i e cells o mic oo ganisms, enhance mass ans e in di e en ope a ions o he ood indus y
(e.g., ex ac ion o in acellula componen s o in e es , dehyd a ion, in usion o compounds in o he
cells, e c.), and modi y ood s uc u e [49,50].
3.1.2. Equipmen
Basic componen s o an appa a us o he applica ion o PEF a e a pulse gene a o and a ea men
chambe . The pulse gene a o is a Ma x gene a o o squa e wa e o m pulses wi h a di ec cu en
powe supply which con e s al e na ing cu en o di ec cu en line ha is used, in u n, o cha ge
a se o capaci o s a high ol age. When he high ol age swi ch (a high-powe solid-s a e swi ch)
is opened, he capaci o s a e cha ged. I he high-powe swi ch is hen closed, all he elec ical
ene gy s o ed in he capaci o s is deli e ed o he ea men chambe . The swi ching sys em pe mi s
Be e ages 2019,5, 47 7 o 18
he con olled discha ge o he capaci o in he o m o pulses o e y sho du a ion a e y high
equencies ( eaching hund eds o pulses pe second).
Du ing PEF p ocessing, a liquid ood o pumpable p oduc is passed h ough a ea men
chambe whe e i is subjec ed o sho pulses o high ol age. The ea men chambe consis s o wo
elec odes made o a conduc ing ma e ial such as s ainless s eel o i anium; hey a e sepa a ed by
an insula ing ma e ial, which o ms an enclosu e con aining he ood ma e ial. Di e en ypes o
ea men chambe s ha e been designed o minimize he e ec o elec olysis as well as co osion.
The wo mos impo an ea men chambe designs ha a e p esen ly conside ed o he comme cial
applica ion o PEF a e pa allel elec ode and co-linea con igu a ions. The la e con igu a ion is he one
habi ually used o p ocessing c ushed g apes a e des emming, wi h he pu pose o elec opo a ing
he cy oplasmic memb ane o g ape skin cells o acili a e he ex ac ion o polyphenols du ing he
mace a ion- e men a ion s age. The co-linea ea men chambe consis s o an elec ically insula ing
ube h ough which he g ape mash lows. The elec odes a e loca ed in he middle (high ol age)
and on ei he side o he chambe (g ound). They consis o wo me al pipes ha also se e as
he en ance and exi o he luid. The ci cula sec ion o his co-linea con igu a ion acili a es
i s ins alla ion in wine y ci cula ion pipes used o anspo c ushed and des emmed g apes o he
e men a ion-mace a ion anks (Figu e 1) [51].
The lack o eliable and iable indus ial-scale equipmen has limi ed he comme cial exploi a ion
o PEF in he ood indus y o many yea s. Howe e , ecen de elopmen s in pulse powe gene a o s
ha e enabled he design o PEF equipmen wi h cha ac e is ics ha can mee indus ial s anda ds in
e ms o eliabili y and wo kloads [52].
Be e ages 2019, 5, x FOR PEER REVIEW 7 o 18
he con olled discha ge o he capaci o in he o m o pulses o e y sho du a ion a e y high
equencies ( eaching hund eds o pulses pe second).
Du ing PEF p ocessing, a liquid ood o pumpable p oduc is passed h ough a ea men
chambe whe e i is subjec ed o sho pulses o high ol age. The ea men chambe consis s o wo
elec odes made o a conduc ing ma e ial such as s ainless s eel o i anium; hey a e sepa a ed by an
insula ing ma e ial, which o ms an enclosu e con aining he ood ma e ial. Di e en ypes o
ea men chambe s ha e been designed o minimize he e ec o elec olysis as well as co osion.
The wo mos impo an ea men chambe designs ha a e p esen ly conside ed o he comme cial
applica ion o PEF a e pa allel elec ode and co-linea con igu a ions. The la e con igu a ion is he
one habi ually used o p ocessing c ushed g apes a e des emming, wi h he pu pose o
elec opo a ing he cy oplasmic memb ane o g ape skin cells o acili a e he ex ac ion o
polyphenols du ing he mace a ion- e men a ion s age. The co-linea ea men chambe consis s o
an elec ically insula ing ube h ough which he g ape mash lows. The elec odes a e loca ed in he
middle (high ol age) and on ei he side o he chambe (g ound). They consis o wo me al pipes
ha also se e as he en ance and exi o he luid. The ci cula sec ion o his co-linea con igu a ion
acili a es i s ins alla ion in wine y ci cula ion pipes used o anspo c ushed and des emmed g apes
o he e men a ion-mace a ion anks (Figu e 1) [51]
The lack o eliable and iable indus ial-scale equipmen has limi ed he comme cial
exploi a ion o PEF in he ood indus y o many yea s. Howe e , ecen de elopmen s in pulse
powe gene a o s ha e enabled he design o PEF equipmen wi h cha ac e is ics ha can mee
indus ial s anda ds in e ms o eliabili y and wo kloads [52].
Figu e 1. Flow cha o g ape p ocessing wi h PEF echnology. (A) des emming; (B) p og essi e ca i y
pump; (C) co-linea ea men chambe ; (D) high ol age elec ode; (E) g ound elec ode; (F)
e men a ion ank.
3.1.3. Impac o he T ea men in he Composi ion o Wine
As compa ed wi h hea ing echniques, o he imp o emen o ex ac ion o polyphenols by PEF
equi es o main ain he solid pa s o he g apes in con ac wi h he liquid phase o di e en pe iods
o ime [53]. The e o e, he e ec o PEF ea men on cell skin en elopes seems o be less agg esi e
Figu e 1.
Flow cha o g ape p ocessing wi h PEF echnology. (
A)
des emming; (
B
) p og essi e
ca i y pump; (
C
) co-linea ea men chambe ; (
D
) high ol age elec ode; (
E
) g ound elec ode; (
F
)
e men a ion ank.
Be e ages 2019,5, 47 8 o 18
3.1.3. Impac o he T ea men in he Composi ion o Wine
As compa ed wi h hea ing echniques, o he imp o emen o ex ac ion o polyphenols by PEF
equi es o main ain he solid pa s o he g apes in con ac wi h he liquid phase o di e en pe iods
o ime [
53
]. The e o e, he e ec o PEF ea men on cell skin en elopes seems o be less agg esi e
han ha o echniques based on hea ing [
54
]. Tes s ca ied ou by di e en au ho s on di e en g ape
a ie ies ag ee ha PEF ea men nei he a ec s he e men a ion p ocess no he physicochemical
p ope ies o he esul ing ed wine. E hanol con en , pH, ola ile acidi y and o al acidi y in he wines
ob ained wi h g apes ea ed by PEF we e simila o con ol wines [53,55].
The elec opo a ion o cell g ape skins by he applica ion o PEF accele a es and inc eases he
ex ac ion o phenolic compounds du ing he mace a ion- e men a ion s age in he ini ica ion o
ed g apes [
56
]. Di e en s udies ha e shown ha , a e he same mace a ion ime han in con ol
wine, PEF ea men ein o ces oenological pa ame e s by a a e o 10% o 60%, depending on he
ex ac ion o polyphenols (colo in ensi y, o al an hocyanin con en , and o al polyphenol con en ) in
he mace a ion- e men a ion s age [53].
Pu
é
olas e al. [
57
] showed ha PEF echnology can help educe mace a ion imes. Cabe ne
Sau ignon wine ob ained om PEF- ea ed g apes (5 kV/cm, 150
µ
s, and 3.67 kJ/kg) p esen ed highe
colo in ensi y, o al an hocyanin con en , and o al polyphenol con en alues, al hough he du a ion o
he mace a ion o he g apes ea ed by PEF was 48 h sho e han o con ol wines. E olu ion du ing
aging o he wine ob ained om g apes ea ed by PEF was simila o con ol wine. The di e ences
in colo in ensi y, o al an hocyanin con en , and o al polyphenol con en obse ed a he end o
e men a ion be ween con ol wine and he wine ob ained om PEF- ea ed g apes we e main ained
a e aging he wine in bo le o oak ba els [
58
]. De e mina ion o indi idual polyphenols by means
o high-pe o mance liquid ch oma og aphy (HPLC) highligh ed ha he wines ob ained by PEF
ea men s did no show di e ences in e ms o he p opo ion o di e en polyphenols, hus indica ing
ha PEF ea men did no selec i ely ex ac phenolic compounds om g ape skins. L
ó
pez-Al a o
e al. [
59
] epo ed ha he con en o es e a ol, one o he mos esea ched phenols in wine due o
i s bene icial p ope ies, inc eased by a p opo ion o 200, 60 and 50% in Temp anillo, Ga nacha and
G aciano, espec i ely, when he g apes we e ea ed wi h PEF be o e mace a ion- e men a ion.
Ene ge ic equi emen s o he elec opo a ion o cells o g ape skins a e lowe han 10 kJ/kg; as a
consequence, he ea men causes an inc emen o less han 2
◦
C in g ape mash empe a u e. This
low impac allows he ob ained wines o main ain hei a ie al cha ac e [
57
]. Some expe imen s
ha e shown ha PEF ea men s encou age he di usion o a oma ic compounds ound in he skin,
as well as o a oma ic p ecu so s [
60
]. PEF ea men did no inc ease he concen a ion o C6 amily
compounds associa ed wi h he baceous a omas in wines ob ained om Ga nacha, Temp anillo, and
G aciano a ie ies [60]. The ea men signi ican ly inc eased mono e penoid compounds, and a had
posi i e e ec on he concen a ion o
β
-ionone, o al es e s, and benzenoid compounds in G enache
wine. Howe e , he ola ile composi ion o Temp anillo and G aciano wines was no a ec ed by PEF.
Senso y analysis did no de ec any d awbacks in Cabe ne Sau ignon wines ob ained wi h g apes
ea ed by PEF. Luengo e al. [
51
] compa ed G enache wines ea u ing simila enological pa ame e s
in e ms o polyphenol con en ob ained, on he one hand, wi h PEF ea ed g apes and 7 days o
mace a ion and, on he o he hand, wi h un ea ed g apes and 14 days o mace a ion. Compa ed
wi h con ol wine, panelis s p e e ed he wine ob ained wi h g apes ea ed by PEF and a sho e
mace a ion pe iod.
3.2. Ul asound
3.2.1. Desc ip ion o he Technique
Acous ic wa es o a speci ic equency lying abo e he de ec ion h eshold o human hea ing (i.e.,
o e 16–18 kHz) a e designa ed as ul asound. Ul asound is di ided in o wo ca ego ies, acco ding
o he equency ange and he in ensi y o ul asonic wa es. The i s g oup, commonly known
Be e ages 2019,5, 47 9 o 18
as high-in ensi y ul asound, ea u es low equency and high in ensi y (20–100 kHz; >10 W/cm
2
).
The second g oup, commonly called diagnos ic ul asound, uses high equency and low powe
(>100 kHz; <1 W/cm2).
When high-in ensi y ul asound passes h ough a liquid medium, a phenomenon called acous ic
ca i a ion occu s [
61
]. Ca i a ion consis s in he implosion o bubbles o med in liquid media when he
local p essu e in he expansion phase alls below apo p essu e. Du ing he implosion, i is es ima ed
ha high empe a u es and p essu es a e eached in e y small spo s and e y sho pe iods o ime:
liquid je s o up o 280 m/s a e likewise gene a ed. These phenomena b ough abou by ca i a ion a e
esponsible o e ec s a ibu ed o high-in ensi y ul asound, such as he inc emen o mass ans e ,
o he b eakage o cells o mic oo ganisms, o o plan o animal issues [
62
]. Ul asound may he e o e
enhance he ex ac ion o polyphenols om he solid pa s o g apes in ed winemaking by b eaking
up he cells, and by acili a ing he di usion o polyphenols om he cells o he mus [63].
3.2.2. Equipmen
An appa a us o he gene a ion o ul asound consis s in a powe supply and a ansduce .
The powe supply con e s al e na ing cu en line ol age o equencies o o e 20 kHz elec ical
ene gy. This high- equency elec ical ene gy is ed o a ansduce , whe e i is con e ed o mechanical
ib a ions a he same equency as he ans o med elec ical cu en . The physical concep unde lying
he ansduce is he piezoelec ic e ec : he p ope y o ce ain ma e ials causes hem o change shape
when an elec ic cu en is applied o hem. An ul asound ansduce con ains a hin disk, squa e, o
ec angle o piezoelec ic ce amic placed be ween wo elec odes which expand and con ac when
subjec ed o al e na ing ol age. The con e e ib a es in a longi udinal di ec ion and ansmi s he
mo ion o he solu ion, he eby causing ca i a ion [61].
A powe ul asound sys em has ecen ly been de eloped o p ocessing des emmed and c ushed
g apes in con inuous low. The equipmen consis s o a hexagonal s ainless-s eel pipe in o which he
ansduce is welded (Figu e 2). The leng h o he pipes con aining he ansduce s is a iable,
depending on he ins alla ion’s p ocessing capaci y, which can each up o en ons pe hou .
The ca i a ion caused by he ul asound ea men p o okes he des uc ion o he cells o he
solid pa s o he g apes, he eby leading o he elease o polyphenols.
Be e ages 2019, 5, x FOR PEER REVIEW 9 o 18
he local p essu e in he expansion phase alls below apo p essu e. Du ing he implosion, i is
es ima ed ha high empe a u es and p essu es a e eached in e y small spo s and e y sho
pe iods o ime: liquid je s o up o 280 m/s a e likewise gene a ed. These phenomena b ough abou
by ca i a ion a e esponsible o e ec s a ibu ed o high-in ensi y ul asound, such as he inc emen
o mass ans e , o he b eakage o cells o mic oo ganisms, o o plan o animal issues [62].
Ul asound may he e o e enhance he ex ac ion o polyphenols om he solid pa s o g apes in ed
winemaking by b eaking up he cells, and by acili a ing he di usion o polyphenols om he cells
o he mus [63].
3.2.2. Equipmen
An appa a us o he gene a ion o ul asound consis s in a powe supply and a ansduce . The
powe supply con e s al e na ing cu en line ol age o equencies o o e 20 kHz elec ical ene gy.
This high- equency elec ical ene gy is ed o a ansduce , whe e i is con e ed o mechanical
ib a ions a he same equency as he ans o med elec ical cu en . The physical concep
unde lying he ansduce is he piezoelec ic e ec : he p ope y o ce ain ma e ials causes hem o
change shape when an elec ic cu en is applied o hem. An ul asound ansduce con ains a hin
disk, squa e, o ec angle o piezoelec ic ce amic placed be ween wo elec odes which expand and
con ac when subjec ed o al e na ing ol age. The con e e ib a es in a longi udinal di ec ion and
ansmi s he mo ion o he solu ion, he eby causing ca i a ion [61].
A powe ul asound sys em has ecen ly been de eloped o p ocessing des emmed and c ushed
g apes in con inuous low. The equipmen consis s o a hexagonal s ainless-s eel pipe in o which he
ansduce is welded (Figu e 2). The leng h o he pipes con aining he ansduce s is a iable,
depending on he ins alla ion’s p ocessing capaci y, which can each up o en ons pe hou . The
ca i a ion caused by he ul asound ea men p o okes he des uc ion o he cells o he solid pa s
o he g apes, he eby leading o he elease o polyphenols.
Figu e 2. Flow cha o g ape p ocessing wi h ul asound echnology. (A) des emming; (B)
p og essi e ca i y pump; (C) ul asound ea men zone; (D) ansduce ; (E) e men a ion ank.
3.2.3. Impac o he T ea men in he Composi ion o Wine
Figu e 2.
Flow cha o g ape p ocessing wi h ul asound echnology. (
A
) des emming; (
B
) p og essi e
ca i y pump; (C) ul asound ea men zone; (D) ansduce ; (E) e men a ion ank.
Be e ages 2019,5, 47 16 o 18
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