Accep ed Manusc ip
Ti le: Mic owa e adia ion and p o ein addi ion modula e
hyd a ion, pas ing and gel heological cha ac e is ics o ice
and po a o s a ches
Au ho s: Ma ina Villanue a, Bea iz De Lamo, Joanna
Ha asym, Felicidad Ronda
PII: S0144-8617(18)30950-0
DOI: h ps://doi.o g/10.1016/j.ca bpol.2018.08.052
Re e ence: CARP 13949
To appea in:
Recei ed da e: 7-6-2018
Re ised da e: 6-8-2018
Accep ed da e: 11-8-2018
Please ci e his a icle as: Villanue a M, De Lamo B, Ha asym J, Ronda F,
Mic owa e adia ion and p o ein addi ion modula e hyd a ion, pas ing and gel
heological cha ac e is ics o ice and po a o s a ches, Ca bohyd a e Polyme s (2018),
h ps://doi.o g/10.1016/j.ca bpol.2018.08.052
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1
Mic owa e adia ion and p o ein addi ion modula e hyd a ion, pas ing
and gel heological cha ac e is ics o ice and po a o s a ches
Ma ina Villanue a1, Bea iz De Lamo1, Joanna Ha asym1, 2, Felicidad Ronda1*
1 Depa men o Ag icul u e and Fo es y Enginee ing, Food Technology, College o
Ag icul u al and Fo es y Enginee ing, Uni e si y o Valladolid, A . Mad id, 44, 34004
Palencia, Spain.
2 Bio-Re Lab, Depa men o Bio echnology and Foods Analysis, Ins i u e o
Chemis y and Food Technology, Facul y o Enginee ing and Economics, W ocław
Uni e si y o Economics, W ocław, Poland.
* Co esponding au ho : [email p o ec ed]; Tel: +34979108339; Fax: +34979108200
G aphical abs ac
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Highligh s:
 The swelling powe o po a o s a ch inc eased a e he mic owa e ea men
 Mic owa e ea men dec eased he swelling powe o ice s a ch
 The p o ein p esence changed he impac o mic owa es on s a ch p ope ies
 Mic owa e adia ion al e ed he pas ing p ope ies o po a o s a ch
 Mic owa e ea men inc eased iscoelas ic moduli o po a o s a ch gels
Abs ac
This s udy e alua ed o i s ime he e ec o Mic owa e (MW) adia ion on sys ems
based on po a o and ice s a ches supplemen ed wi h 5% o calcium caseina e (CA) o
soy p o ein isola e (SPI). The goal o his ea men was he physical modi ica ion o
hese s a ch-based sys ems o p o ide ing edien s o new unc ionali ies. The hyd a ion
and pas ing p ope ies as well as gel iscoelas ic ea u es we e e alua ed. Dynamic
oscilla o y heological es s we e used. The e ec o MW ea men (MWT) depended
on he s a ch bo anical o igin and was signi ican ly a ec ed by p o ein p esence and
ype. MWT o s a ch+p o ein blends e ealed he mos no able changes when SPI was
added. Adding i o ice s a ch dec eased swelling powe (-45%), al e ed iscome ic
p o iles and ein o ced gel s uc u e wi h impo an inc eases in bo h iscoelas ic
moduli (+160%-G’ and +58%-G’’). In blends wi h po a o s a ch, MWT inc eased wa e
abso p ion capaci y (+115%), and dec eased wa e solubili y index (-82%). MWT o
p o ein-po a o blends p omo ed gel s abili y, dec eased hei pas ing p o iles and
esul ed in enhanced iscoelas ic moduli (+483-G’ and 243%-G’’). MWT combined
wi h p o ein addi ion allows designing s a ch-based oods wi h ailo ed p ope ies.
Keywo ds: Mic owa e; unc ional p ope ies; gel heology; po a o s a ch; ice s a ch
1. In oduc ion
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The na i e o m o s a ch has limi ed applica ions in he ood indus y and is o en
modi ied physically, chemically o enzyma ically o imp o e unc ional p ope ies.
Physically modi ied s a ch is conside ed a na u al ma e ial and a highly sa e ing edien ,
much sa e and mo e p edic able han chemically modi ied s a ch. So a , he usage o
physically modi ied s a ch in ood is no es ic ed by legisla ion, which is conside ed
an ou s anding ad an age compa ed o chemically- o enzyma ically-modi ied s a ch
applica ions (Klein e al., 2013).
S a ch, as a mac o-cons i uen o many aw ma e ials and p ocessed oods, con ibu es
o hei nu i ional and unc ional cha ac e is ics, as well as plays he pi o al ole as a
ex u e modi ie (Walkens öm and He mansson, 1998) and in luences many quali y
a ibu es o p ocessed oods (Chen e al., 2015). Rice and po a o s a ches a e among he
mos commonly used o ex u e de elopmen , pa icula ly in glu en- ee (GF) p oduc s.
Du ing mic owa e (MW) adia ion exposu e, ene gy is deli e ed di ec ly o he
ma e ial, hea ing he en i e sample in bulk a a as e hea ing a e han con en ional
hea ing (Bilbao-Sáinz e al., 2007). The e o e, MW adia ion-induced hea ing is sui able
o modi ying s a ches physically and imp o ing hei unc ional p ope ies. In
mois ened s a ches, he in e molecula s uc u e changes om MW-deli e ed ene gy
lead o changes in wa e abso p ion abili y, solubili y and swelling powe , as well as in
s a ch gela iniza ion, syne esis and pas e iscosi y (B aso eanu and Nem anu, 2014).
The e a e many ac o s ela ed o bo h s a ch cha ac e is ics (bo anical o igin, wa e
con en , densi y, dielec ic p ope ies, e c.) and mic owa e i adia ion condi ions
( equency, powe and exposu e ime) ha signi ican ly a ec how g anula s a ch
esponds o MW ea men (MWT) (B aso eanu and Nem anu, 2014). Za a eze e al.
(2010) showed ha mic owa e i adia ion a ec ed he pas ing p ope ies o high-
amylose ice s a ches mo e in ensely han in hose wi h lowe amylose con en .
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Ve meylen e al. (2006) concluded ha highe ea men empe a u es and mois u e
con en s gene ally caused he la ges changes in s a ches. As applied empe a u e and
mic owa e powe le el inc eased, he peak iscosi y o po a o s a ch dec eased (Nadiah
e al., 2015), while he gela iniza ion empe a u e inc eased (Nadiah e al., 2015;
Ve meylen e al., 2006). Villanue a e al. (2018a) concluded ha he MW adia ion
abso p ion capaci y o ice lou exhibi ed a posi i e quad a ic ela ionship wi h wa e
con en , while he lou d y ma e did no show any adia ion abso p ion capaci y. They
also obse ed changes in physical p ope ies o ea ed ice lou , such as ice lou
pa icle mo phology o he c ys allini y/amo phous egion a io.
Beyond physico-chemical and unc ional conside a ions, s a chy glu en- ee oods a e
o en conside ed nu i ionally poo compa ed o whea -based coun e pa s due o hei
de icien p o ein/s a ch a io (Villanue a e al., 2015). The addi ion o p o ein is a iable
al e na i e o imp o e he nu i ional p ope ies o hese p oduc s. P o eins a e also
known o modi y s uc u e and ex u e in oods (Villanue a e al., 2018b). Soy p o eins
isola e (SPI) and calcium caseina e (CA) ha e been in es iga ed in GF applica ions
because o hei e ec on nu i ion and s uc u e o p oduc s (K upa-Kozak e al., 2011;
Ronda e al., 2014). S a ch-p o ein in e ac ions a ec he heological, pas ing,
gela iniza ion, ex u al and physicochemical p ope ies o ood sys ems (Gallaghe e
al., 2003; Ronda e al., 2011; Ronda e al., 2014; Villanue a e al., 2015). As a as we
know, how MWT a ec s p o ein-s a ch mix u es has no been s udied so a , in spi e o
how impo an endogenous p o eins seem o be in he e ec o hea -mois u e ea men
(HMT) on ea ed- lou p ope ies (Puncha-a non and U apap, 2013). As p o ein and
s a ch blends can se e as model sys ems o he s udy o mo e complex ma ices such
as lou , he impac o hei in e ac ions on changes appea ing in MW ea ed samples
needs o be be e unde s ood. The main objec i e o his s udy was o in es iga e he
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combined e ec o mic owa e adia ion and p o ein addi ion (5% CA o SPI) on he
hyd a ion, pas ing p ope ies and gel iscoelas ic p ope ies o po a o and ice s a ches..
The ul ima e goal o his s udy is o p o ide s a ch-based ing edien s wi h ailo ed
unc ional p ope ies.
2. Ma e ial and me hods
2.1. Ma e ials
Rice s a ch and po a o s a ch we e supplied by Fe e Alimen ación S.A. (Ba celona,
Spain). Soybean p o ein isola e (SPI) Sup o 500-E IP (pu i y ~ 90%) was ob ained om
P o eedo a hispano-holandesa S.A. (Ba celona, Spain), and calcium caseina e (CA)
(pu i y ~ 96%) om A mo P o éines (Sain -B ice-en-Coglès, F ance). P o ein added in
all he s a ch+p o ein blends was 5 g/100 g blend. Twel e di e en samples we e
in es iga ed: na i e and MW- ea ed ice and po a o s a ches alone and blended wi h 5
g/100 g o CA o SPI p o eins.
2.2. Mic owa e ea men o s a ch samples
Samples we e ea ed by MW adia ion a ini ial mois u e con en o 30%. The l
mois u e con en o he samples was measu ed ollowing he Ame ican Associa ion o
Ce eal Chemis s (AACC) 44–19 me hod and he wa e necessa y o adjus o 30%
mois u e con en was sp ayed on o he samples while hey we e mixed in a Bea Teddy
Mixe (Bea 5L Teddy, Swansea, UK) o 15 min. Samples we e hen he me ically
sealed in bags and held 24 h a 4 ± 2°C o he mois u e equilib a ion.
Hyd a ed samples (100.0 g each) we e hea ed in a SHARP R-342 (Osaka, Japan)
mic owa e o en in a cylind ical polye hylene con aine closed wi h needle-punched,
mic owa e-sa e ood g ade ilm. The homogenous dis ibu ion o adia ion on he
sample is o c ucial impo ance. So, he con aine was s i ed cons an ly o ensu e a
uni o m ene gy and empe a u e dis ibu ion du ing ea men as desc ibed in
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Villanue a e al. (2018b). MW adia ion equency was 2450 MHz and he powe ,
900W. MW was applied in cycles o 20 s adia ion wi h 40 s down ime, o a o al o 32
min and 640 s o MWT; a e wa ds, samples we e le o cool o 5 minu es.
Agglome a es o med du ing ea men we e manually disin eg a ed in a labo a o y
mo a o < 0.5 mm. Sample empe a u e du ing MWT was measu ed wi h Tes o e m
he mome e s ips o di e en scales, and 0.5ºC accu acy (Ins umen os Tes o S.A.,
Ba celona, Spain), as desc ibed in Villanue a e al. (2018a). Each measu emen was
made in duplica e. The empe a u e e olu ion cu es ob ained o he po a o and ice
s a ch sys ems s udied we e equi alen o hose epo ed o 30% mois ened- ice lou
in ou p e ious wo k (Villanue a e al., 2018a). The empe a u e eached and
main ained a e 4 min o ea men was 157±5ºC o all samples.
2.3. Amylose con en
The amylose con en o he s a ch samples was de e mined using a colo ime ic assay
ki o amylose/amylopec in a io de e mina ion in s a ch (Megazyme In e na ional
I eland L d., I eland), acco ding o he manu ac u e ’s p ocedu e based on Gibson e al.,
(1997) me hod. The me hod de e mines he soluble amylose in ace a e/sal solu ion
suscep ible o α-amylase/ amyloglucosidase diges ion. Each sample was analysed a
leas in duplica e.
2.4. Hyd a ion p ope ies
Wa e abso p ion capaci y (WAC) o he samples was de e mined by he cen i uga ion
me hod desc ibed by Abebe e al., (2015). Two g ams o sample (ws) we e mixed wi h
20 mL o dis illed wa e in 50 mL cen i uge ubes. The dispe sions we e hold a oom
empe a u e o 30 min wi h occasionally o exed (Heidolph Reax, Schwabach,
Ge many) and ollowed by cen i uga ion o 30 min a 3000×g (The mo Fishe
Scien i ic, Wal ham, USA). The supe na an was emo ed and weighed (ws+w) and
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esul s we e exp essed as g ams o wa e e ained pe g am o sample. Wa e abso p ion
index (WAI) and wa e solubili y index (WSI) we e measu ed wi h sligh modi ica ion
o he me hod used by Abebe e al. (2015). Each sample o 2.5 g (w0) was dispe sed in
30 ml o dis illed wa e in a ed cen i uge ubes. A e cooking o 10 min in a 90ºC
wa e ba h, samples we e cooled o oom empe a u e and cen i uged a 4000xg o 10
min. The supe na an was pou ed in o a p e-weighed e apo a ing capsule o de e mine
he solid con en and he sedimen was weighed (wss). The weigh o soluble solids was
eco e ed by e apo a ing he supe na an o e nigh a 110ºC (wds). WAC, WAI, WSI
and swelling powe (SP) we e calcula ed using he ollowing equa ions:
𝑊𝐴𝐶(𝑔
𝑔) = 𝑤𝑠+𝑤−𝑤𝑠
𝑤𝑠 (1)
𝑊𝐴𝐼 (𝑔
𝑔) = 𝑤𝑠𝑠
𝑤0 (2)
𝑊𝑆𝐼( 𝑔
100𝑔) = 𝑤𝑑𝑠
𝑤0×100 (3)
𝑆𝑃(𝑔
𝑔) = 𝑤𝑠𝑠
(𝑤0−𝑤𝑑𝑠) (4)
All esul s we e e e ed o d y ma e o a oid he e ec o di e en wa e con en in
he samples.
2.5. The mo iscous es : Viscome ic p o ile
Following AACC In e na ional Me hod 76-21.01 S anda d 2, iscome ic p o iles o
MW- ea ed and un ea ed samples we e ob ained using a Kinexus P o+ heome e
(Mal e n Ins umen s L d., Mal e n, UK) supplied wi h s a ch pas ing cell and
con olled by Space so wa e. Samples o ice s a ch (3 g, 14% mois u e-based) o
po a o s a ch (2 g, 14% mois u e-based) wi h o wi hou added p o ein we e ans e ed
o a canis e whe e 25 mL±0.1 mL o dis illed wa e was added. Each s a ch suspension
was equilib a ed a 50 ºC o 1 min, hea ed o 95 ºC a a a e o 6 ºC/min, main ained a
95 ºC o 5 min, hen cooled o 50 ºC a a a e o 6 ºC/min, and main ained a 50 ºC o 2
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min. Paddle speed was se a 960 pm o he i s 10 s and hen 160 pm o he es o
he analysis. Each sample was analyzed a leas in duplica e. Pa ame e s calcula ed om
he pas ing p o iles we e pas ing empe a u e (PT), peak iscosi y (PV), ough iscosi y
(TV), b eakdown (BD), inal iscosi y (FV), se back (ST) and peak ime.
2.6. Rheological measu emen s
Dynamic oscilla o y es s o he wel e po a o and ice s a ch gels we e pe o med wi h
Kinexus P o+ heome e (Mal e n Ins umen s L d., Mal e n, UK) wi h pa allel pla e
geome y (40 mm diame e ) o se a ed su ace and wi h 1 mm o wo king gap. The gel
samples we e made ollowing he same p ocedu e desc ibed o he mo iscous es s.
Di e en concen a ions o ice and po a o s a ches we e used o p epa e hei gels, so
ha he gels p oduced om na i e s a ches we e o simila consis encies. Jus a he end
o he pas ing es he gel was emo ed om he canis e and placed be ween he pla es,
he sample excess was emo ed and he sample was le o es o 5 min o allow
elaxa ion. Tempe a u e was s abilized a 25ºC wi h a Pel ie pla e con olle . S ess
sweeps we e pe o med om 0.1 o 500 Pa a a cons an 1 Hz equency. F equency
sweeps we e ca ied ou om 10 o 1 Hz in he linea iscoelas ic egion (a a cons an
alue o 1 Pa). F equency sweep da a we e i ed o po en ial equa ions as desc ibed by
Ronda e al. (2014). All gels we e p epa ed a leas in duplica e and heological es s
we e also pe o med in duplica e.
2.7. S a is ical analysis
S a g aphics Cen u ion .6 (Bi s eam, Camb idge, MN, USA) was used o mul i ac o
analysis o a iance (ANOVA) o he da a. The Leas Signi ican Di e ence es was
used o e alua e signi ican di e ences (p < 0.05) be ween samples.
3. Resul s and discussion
3.1. Amylose con en
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high humidi y hea ing pe iod wi h a d y hea ing pe iod could explain he in e media e
beha iou ha we obse ed in ice s a ch wi h espec o ha ob ained by Puncha-a non
and U apap (2013) and Qiu e al. (2016).
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Table 2. E ec o mic owa e ea men and p o ein p esence on he pas ing p ope ies o s a ch samples.
S a ch
P o ein
MW
ea men
PT(ºC)
PV (Pa·s)
TV ( Pa·s )
BD ( Pa·s )
FV ( Pa·s )
ST ( Pa·s )
Peak ime (s)
Rice
0
0
74.6
ab
2.55
c
1.25
c
1.30
b
2.24
c
0.99
d
665
d
1
73.9
a
2.73
d
1.23
c
1.49
c
2.29
c
1.06
e
627
ab
CA
0
75.8
b
2.09
a
1.02
a
1.08
a
1.82
a
0.80
b
650
c
1
74.6
ab
2.27
b
1.11
b
1.16
a
2.00
b
0.88
c
635
b
SPI
0
73.9
a
2.32
b
1.03
a
1.30
b
1.72
a
0.69
a
618
a
1
74.3
a
2.68
cd
1.35
d
1.33
b
2.31
c
0.96
d
620
a
Analysis o a iance and signi icance (p- alues)
Fac o 1 (p o ein ype)
*
***
***
***
***
***
***
Fac o 2 (MW ea men )
ns
***
***
**
***
***
***
Fac o 1x2
ns
ns
***
ns
***
***
**
Po a o
0
0
70.0
a
6.88
e
2.06
b
4.82
d
2.84
ab
0.78
a
358
a
1
76.2
b
1.85
b
2.11
b
0.00
a
3.70
b
1.59
b
687
c
CA
0
71.4
a
3.15
d
1.66
a
1.49
c
2.34
a
0.67
a
518
b
1
79.0
c
1.60
a
1.59
a
0.00
a
2.43
a
0.84
a
860
d
SPI
0
70.5
a
2.72
c
1.50
a
1.22
b
2.39
a
0.89
ab
538
b
1
78.1
c
1.92
b
2.01
b
0.00
a
3.29
ab
1.28
ab
762
cd
Analysis o a iance and signi icance (p- alues)
Fac o 1 (p o ein ype)
**
***
**
***
*
ns
**
Fac o 2 (MW ea men )
***
***
*
***
*
*
***
Fac o 1x2
ns
***
**
***
ns
ns
ns
P o ein: 0: wi hou p o ein; CA: 5% calcium caseina e; SPI: 5% soy p o ein isola e. MW ea men : 0: wi hou ea men ; 1: wi h ea men .
PT: pas ing empe a u e, PV: peak iscosi y, TV: ough iscosi y, BD: b eakdown, FV: inal iscosi y, ST: se back.
The di e en le e s in he co esponding column wi hin each s a ch ype indica e s a is ically signi ican di e ences be ween means a p<0.05.
Analysis o a iance and signi icance: *** p<0.001. ** p<0.01. * p<0.05. ns: no signi ican .
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0
10
20
30
40
50
60
70
80
90
100
0
0,5
1
1,5
2
2,5
3
0 200 400 600 800 1000 1200 1400
Tempe a u e (ºC)
Viscosi y (Pa·s)
Time (s)
0
10
20
30
40
50
60
70
80
90
100
0
1
2
3
4
5
6
7
8
0 200 400 600 800 1000 1200 1400
Tempe a u e (ºC)
Viscosi y (Pa·s)
Time (s)
Figu e 1. Pas ing p o iles o na i e and modi ied samples o ice s a ch (A) and po a o
s a ch (B). S a ch wi hou p o ein a e ep esen ed by , wi h 5% calcium caseina e
by , and wi h 5% soy p o ein isola e by . Mic owa e- ea ed p o ein- ee
samples a e ep esen ed by , mic owa e- ea ed samples wi h 5% calcium
caseina e by and wi h 5% soy p o ein isola e by . The empe a u e p o ile is
ep esen ed by in he second axis.
B)
A)
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The low peak ime o na i e po a o s a ch inc eased signi ican ly wi h MWT, eaching
he peak alues o ice s a ch. Na i e po a o s a ch showed a much highe BD han ice
s a ch, 4.82 e sus 1.30 Pa·s (Table 2). A low BD alue means high pas e s abili y
e sus hea and shea , and is ela ed o g anule igidi y and high lipid con en (Singh e
al., 2003). Adding p o ein inc eased he pas e s abili y o po a o s a ch, dec easing he
BD alue by 69% and 75% o CA and SPI, espec i ely. Howe e , in he case o ice
s a ch, only CA had an e ec and i was much sligh e . MWT ha dly a ec ed he BD
alue o he ice s a ch samples. Howe e , he ea men did educe po a o s a ch BD o
ze o, ega dless o he p esence o p o eins. This e lec s he abili y o MWT o inc ease
po a o s a ch gel s abili y e sus hea ing and shea ing. This beha iou has al eady been
obse ed in ube s a ches (po a o and apioca) when hey we e mois ened and he mally
modi ied ei he by con ec ion hea ans e (Klein e al., 2013) o MW adia ion
(Nadiah e al., 2015). ANOVA analysis (Table 2) e ealed ha p o ein ype and MW
ea men had a singula ly s ong e ec on PT, PV and BD alues o ice s a ch, wi hou
any mu ual in e ac ion o hese ac o s. Howe e , po a o s a ch was highly suscep ible,
no only o p o ein ype supplemen a ion o MWT bu also o he double in e ac ion o
he wo ac o s on PV and BD.
The inal iscosi y (FV) o he gels o med a e subsequen cooling dec eased in
p esence o ei he animal o ege al p o ein. The same had p e iously been epo ed by
Ronda e al. (2014). The e ec was mo e p onounced o ice s a ch gels han o po a o
gels. MWT inc eased he FV o ice+p o ein gel, while po a o s a ch FV was
una ec ed. ANOVA analysis showed ha each o he main ac o s in luenced inal
iscosi y, bu he double e ec was ound only in ice s a ch.
Se back iscosi y (ST), mainly ela ed o he amylose leached om s a ch g anules and
hei endency o eo ganise a e gela inisa ion (Miles e al., 1985), was equi alen o
bo h na i e s a ches in ou s udy. Adding p o eins signi ican ly (p<0.001) dec eased his
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alue in ice s a ch blends, while po a o alues emained una ec ed. MWT inc eased
ST in all ice s a ch-based o mula ion and p o ein- ee po a o s a ch. In he case o
po a o s a ch he inc ease was om 0.78 o 1.59 Pa·s. The ANOVA esul s con i med
ha he simple and double- ac o e ec s on ice s a ch ST we e highly signi ican (p <
0.001). Howe e , excep o MWT, he e we e almos no e ec s on po a o s a ch.
3.4. Gel heological p ope ies
The iscoelas ic p ope ies o gel samples we e s udied by dynamic oscilla o y es s,
applying a sinusoidal s ess o he samples. The ob ained iscoelas ic moduli as a
unc ion o s ess a e shown in Figu e 2. The pa ame e s ob ained om i ing he
mechanical spec a o powe law a e summa ized in Table 3. The high R2 alues
ob ained, always abo e 0.998 o all samples, demons a e ha he sys ems s udied
adjus ed well o he powe law model. S ess sweeps made i possible o es ablish he
linea iscoelas ic egion (LVR) by iden i ying he maximum s ess (τmax) ha samples
could ole a e conse ing hei s uc u e. These alues a e also included in Table 3. The
elas ic modulus, G’, o gels made om un ea ed s a ches s a ed o dec ease a 108 and
267 Pa o ice and po a o s a ch, espec i ely. This indica ed ha s a ch sample
s uc u es had di e en esis ances o s ess-induced up u e. Acco ding o Villanue a e
al. (2018b), po a o gels p ese ed hei s uc u e beyond ice s a ch gels, meaning ha
po a o gels had good mechanical esis ance and ha dness. Adding p o eins o ice s a ch
gels educed hei esis ance o b eakage and led o alues o τmax 36% and 49% lowe
han hose o p o ein- ee gels o CA and SPI, espec i ely. The e ec on po a o s a ch
gels di e ed depending on p o ein ype: CA dec eased τmax by 19%, while SPI inc eased
i by 14%, compa ed o p o ein- ee po a o gels. MWT inc eased he s eng h o ice
gels, al hough his e ec was only signi ican in SPI-en iched ice gels. The e ec o
MW adia ion on po a o gels was he opposi e and s onge , being much mo e
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ema kable in he p esence o p o eins: τmax dec eased by 60% in gels wi hou p o ein,
88% in CA p o ein-en iched gels and 72% in SPI-en iched gels.
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Figu e 2. S ain sweeps o ice (A) and po a o (B) s a ch gels wi hou p o ein addi ion (A1 and B1), wi h 5% calcium caseina e (A2 and B2) o 5%
soy p o ein isola e (A3 and B3). Non- ea ed samples a e ep esen ed wi h ci cles and MW- ea ed samples wi h iangles. The elas ic modulus, G’,
is ep esen ed using solid symbols and he iscous modulus, G’’, wi h open symbols.
Rice A1
1
10
100
1000
0,1 110 100 1000
G', G'' (Pa)
S ess (Pa)
1
10
100
1000
0,1 1 10 100 1000
G', G'' (Pa)
S ess (Pa)
Po a o B1
B2
A3
A2
B3
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Table 3. E ec o mic owa e ea men and p o ein p esence on he dynamic heological cha ac e is ics o s a ch gels.
S a ch
P o ein
MW
ea men
G’1 (Pa)
a
G’’1 (Pa)
b
( an δ)1
c
max (Pa)
Rice
0
0
52
ab
0.17
b
14
ab
0.36
a
0.27
bc
0.18
ab
108
c
1
74
bc
0.15
a
17
ab
0.35
a
0.23
ab
0.20
b
119
c
CA
0
41
a
0.18
b
12
a
0.36
a
0.29
c
0.18
a
69
ab
1
47
a
0.17
b
13
ab
0.35
a
0.27
bc
0.18
ab
85
b
SPI
0
39
a
0.19
b
12
a
0.36
a
0.30
c
0.17
a
55
a
1
88
c
0.14
a
19
b
0.35
a
0.22
a
0.20
b
105
c
Analysis o a iance and signi icance (p- alues)
Fac o 1 (p o ein ype)
ns
ns
ns
ns
ns
ns
**
Fac o 2 (MW ea men )
**
**
*
ns
**
*
**
Fac o 1x2
ns
ns
ns
ns
ns
ns
*
Po a o
0
0
53
a
0.181
e
16
a
0.29
b
0.31
e
0.11
ab
267
d
1
421
d
0.077
a
51
b
0.22
a
0.12
a
0.15
c
101
b
CA
0
46
a
0.184
e
14
a
0.28
b
0.31
e
0.10
a
216
c
1
171
b
0.141
c
39
b
0.28
b
0.23
c
0.14
bc
26
a
SPI
0
53
a
0.164
d
14
a
0.28
b
0.27
d
0.12
abc
304
d
1
309
c
0.099
b
48
b
0.23
a
0.15
b
0.14
bc
84
b
Analysis o a iance and signi icance (p- alues)
Fac o 1 (p o ein ype)
***
***
ns
*
***
ns
**
Fac o 2 (MW ea men )
***
***
***
**
***
**
***
Fac o 1x2
***
***
ns
*
***
ns
ns
P o ein: 0: wi hou p o ein; CA: 5% calcium caseina e; SPI: 5% soy p o ein isola e. MW ea men : 0: wi hou ea men ; 1: wi h ea men .
G’1, G’’1, and ( an δ)1, ep esen he elas ic and iscous moduli and he loss angen a a equency o 1 Hz. The a, b and c exponen s quan i y he dependence deg ee o dynamic
moduli and he loss angen wi h he oscilla ion equency, ω. τmax ep esen s he maximum s ess ha GF ma ices can ole a e in he LVR.
The di e en le e s in he co esponding column wi hin each s a ch ype indica e s a is ically signi ican di e ences be ween means a p<0.05.
Analysis o a iance and signi icance: *** p<0.001. ** p<0.01. * p<0.05. ns: no signi ican .
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In all samples, G’ and G” inc eased wi h equency, G’ being highe han G”, which
indica ed ha elas ic cha ac e p e ailed o e iscous ac o (Xie e al., 2013). The gel
iscoelas ic moduli G1’ and G1’’ and he loss angen a 1 Hz we e he same o he wo
na i e s a ches (po a o and ice) ega dless o p o ein p esence o ype. Howe e , MWT
always inc eased gel consis ency, his impac was dependen on p o ein p esence and
ype and s a ch ype. The e ec o MWT on gel heological p ope ies was also s ongly
dependen on he bo anical o igin o he s a ches. Bo h iscoelas ic moduli inc eased in
MW- ea ed po a o s a ch samples, while only SPI addi ion a ec ed he moduli in
ea ed ice ones. The G1’ and G1’’ inc eases om MWT o po a o gels we e ela ed o
he gela iniza ion le el and he ne wo k de elopmen o chains leached om he s a ch
g anules du ing he MW ea men (Xie e al., 2013). As a consequence o MWT, he
inc ease in he elas ic modulus was always mo e no iceable han in he iscous
modulus. Consequen ly, MWT was always accompanied by a d op in he loss angen ,
which mean a ein o cemen o he elas ic cha ac e o gels made om MW- ea ed
s a ches o s a ch+p o ein blends compa ed o he un ea ed coun e pa s. Simila esul s
ha e been epo ed o he mally-modi ied po a o s a ch (G yszkin e al., 2014). Tan δ
alue was e y low o all samples (always < 0.4), which co esponded wi h a well
c oss-linked ne wo k. Adding SPI dec eased he gel loss angen o po a o s a ch, bu
ice gels we e una ec ed by p o ein addi ion. Villanue a e al. (2018b) epo ed ha he
heological p ope ies o ice and po a o s a ch gels o i ied wi h egg albumen o SPI
depended signi ican ly on he double in e ac ion (s a ch x p o ein), concluding ha he
same p o ein exe ed a di e en e ec depending on he s a ch sou ce. The alues o he
“a” exponen we e always below he exponen “b”. This mean ha G’’ inc eased wi h
equency as e han G’. This yielded an inc ease in an  wi h equency, which could
also be con i med om he posi i e alue o exponen “c” (Table 3). Consequen ly, gels
inc eased hei iscous beha iou wi h equency.
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4. Conclusions
MWT has been shown o be success ul in modi ying s a ches and hei p o ein mix u es
and has p o en o be e icien in al e ing he physical p ope ies o p oduc s made om
hem. I is pa icula ly use ul in he de elopmen o p o ein-en iched GF p oduc s. The
esul s showed ha he e ec o MWT is dependen on s a ch sou ce and p o ein ype.
MWT changed he hyd a ion p ope ies and enhanced wa e abso p ion index and
swelling powe in po a o samples, while i dec eased hem in ice s a ch samples,
ega dless o ype o p o ein added. MW adia ion in luenced mainly he pas ing
p ope ies o po a o s a ch, inc easing pas ing empe a u e and se back, and dec easing
peak iscosi y and b eakdown. The ea men inc eased he iscoelas ic moduli
signi ican ly and dec eased he loss angen o po a o gels bo h wi h and wi hou
p o eins. In gene al, he inclusion o p o eins inc eased WAC, WAI, SP and WSI,
dec eased he iscosi y o gels and inc eased hei s abili y, being he e ec mo e
ma ked o SPI inco po a ion.
This esea ch helps unde s and he changes ha occu du ing MW ea men s o model
sys ems and can help design and imp o e he quali y o new p oduc s. MW-assis ed
hea ing is an inno a i e lou ea men me hod and can be used as an al e na i e o
chemical modi ica ion.
Acknowledgemen s
The au ho s a e g a e ul o he inancial suppo o he Spanish Minis e io de Economía
y Compe i i idad and he Eu opean Regional De elopmen Fund (FEDER) (AGL2015-
63849-C2-2-R) and o he Jun a de Cas illa y León/FEDER VA072P17. Joanna
Ha asym (MSCA) hanks he Eu opean Commission o he Indi idual Fellowship
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