Depósi o de in es igación de la Uni e sidad de Se illa
h ps://idus.us.es/
“This is an Accep ed Manusc ip o an a icle published by Else ie in LWT-
FOOD SCIENCE AND TECHNOLOGY on 2013, a ailable
a : h ps://doi.o g/10.1016/j.lw .2012.04.021.”
Employmen o di e en p ocesses o he p oduc ion o s awbe y inega s:
1
E ec s on an ioxidan ac i i y, o al phenols and monome ic an hocyanins
2
C. Ubeda1, R. Callejón1, C. Hidalgo2, M.J. To ija2, A.M. T oncoso1, M.L. Mo ales1*
3
1Á ea de Nu ición y B oma ología. Facul ad de Fa macia. Uni e sidad de Se illa.
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C/ P. Ga cía González nº2, E- 41012. Se illa. Spain.
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2Depa amen o de Bioquímica y Bio ecnología. Facul ad de Enología. Uni e si a
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Ro i a i Vi gili. C/ Ma cel·lí Domingo s/n. E- 43007. Ta agona. Spain.
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*Co esponding au ho : [email p o ec ed]; Tel.: 34-954-556760; Fax.: 34-954-233765
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Abs ac
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The use o s awbe y su pluses o he p oduc ion o added alue p oduc s seems o be
10
a good solu ion choice o a oid he was e o his ui . We p oduced s awbe y inega s
11
h ough double e men a ion (alcoholic and ace ous) om h ee di e en ha es s o
12
F aga ia x ananassa a . Cama osa. The objec i e was o s udy he e olu ion o
13
an ioxidan ac i i y, o al phenols and monome ic an hocyanins du ing he inega
14
p oduc ion p ocess. These pa ame e s inc eased when sulphu dioxide and pec oly ic
15
enzymes we e added o subs a es. Inocula ion wi h he Saccha omyces ce e isiae s ain
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RP1 p oduced wines wi h hal he an hocyanins wi h espec o he spon aneous
17
e men a ions. The use o wood ba els, pa icula ly che y wood ba els, had a posi i e
18
e ec on all he pa ame e s de e mined. All measu ed pa ame e s dec eased du ing he
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double e men a ion p ocess. In gene al, he ace i ica ion s age led o a high loss o
20
an ioxidan compounds. Mo eo e , he p oduc ion o hese inega s a a semi-pilo scale
21
yielded inal commodi ies wi h he bes alues o an ioxidan ac i i y, o al phenols and
22
monome ic an hocyanins compa ing wi h he inega s ob ained in 2008 and 2009
23
ha es .
24
Keywo ds: an ioxidan ac i i y; monome ic an hocyanins; s awbe y; inega ; wine.
25
2
1. In oduc ion
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S awbe ies a e a widely esea ched ui o hei nu i ional and heal h bene i s as
27
well as hei o ganolep ic p ope ies. This ui is ich in i amins, mine als, ib e and
28
phy ochemicals. In addi ion, s awbe ies con ain po en ially bioac i e compounds and
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a e a g ea sou ce o phenolic compounds such as la onoids and phenolic acids (Aaby,
30
K., Sk ede, G., & W ols ad, R. E, 2005; Mää ä-Riihinen, K. R., Kamal-Eldin, A., &
31
Tö önen, A. R, 2004; See am, N. P., Lee, R., Scheulle , H. S., & Hebe , D, 2006). All
32
o hese phenolic compounds ha e been shown o p e en oxida i e p ocesses,
33
pa icula ly hose caused by eac i e oxygen species (ROS) (Aaby, K., Ekebe g, D., &
34
Sk ede, G, 2007; Ce ezo, A. B., Cue as, E., Win e hal e , P., Ga cia-Pa illa, M. C., &
35
T oncoso, A. M, 2010a). These compounds make s awbe ies a highly an ioxidan ui
36
(Aaby e al., 2005; Wol e, K. L., Kang, X., He, X., Dong, M., Zhang, Q., & Liu, R. H,
37
2008) wi h po en ial heal h bene i s. Among he nume ous heal hy p ope ies desc ibed
38
in he li e a u e a e an i-p oli e a i e e ec s on cance cells (Meye s, K. J., Wa kins, C.
39
B., P i s, M. P., & Liu, R. H, 2003; Olsson, M. E., Ande sson, C. S., O edsson, S.,
40
Be glund, R. H., & Gus a sson, K, 2006) and he an ioxidan and an i-in lamma o y
41
e ec s ha ha e been shown o educe ca dio ascula disease isk ac o s in se e al
42
p ospec i e coho s udies (Hannum, 2004).
43
Acco ding o he la es da a om he FAO (FAOS a , FAO 2011), Spain is he second-
44
la ges s awbe y p oduce in he wo ld; a la ge po ion o his p oduc ion is ha es ed
45
in Huel a (Andalucía). E e y yea , pa o he c op is disca ded o a ious easons,
46
including size o de o ma ions o he be ies, o o e p oduc ion which leads o
47
su pluses. Because inega is gene ally an inexpensi e p oduc , i s p oduc ion equi es
48
low-cos aw ma e ials, such as sub-s anda d ui and seasonal ag icul u al su pluses
49
(Solie i & Giudici, 2009). In addi ion, he e is a g owing demand o ui inega s,
50
3
which a e sold as a heal h ood (Shau-mei & Chang, 2009). The use o s awbe ies o
51
second quali y, which a e s ill sui able o human consump ion, o p oduc ion heal hy
52
inega s wi h special o ganolep ic nuances may be a good me hod o educe losses due
53
o disca ding he ui .
54
Fo his pu pose, we ha e p oduced s awbe y inega s using second-quali y
55
s awbe ies employing wo-s age e men a ion and assessed di e en condi ions and
56
ea men s. The aim o his wo k was o e alua e he changes in he an ioxidan ac i i y
57
(AA), o al phenols index (TPI) and o al monome ic an hocyanins (TA) du ing he
58
p oduc ion p ocess o s awbe y inega . In addi ion, an adequa e ex ac ion me hod o
59
pe o m hese de e mina ions was designed.
60
2. Ma e ials and me hods
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2.1. Chemicals
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The eagen s ace one, me hanol, Folin-Ciocal eu eagen , e hanol, di-po assium
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hyd ogen phospha e (anhyd ous), sodium di-hyd ogen phospha e 1-hyd a e, po assium
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chlo ide, sodium ace a e and sodium ca bona e (anhyd ous) we e pu chased om Me ck
65
(Da ms ad , Ge many). Fluo escein sodium and gallic acid we e supplied om Fluka
66
(Mad id, Spain). 6-hyd oxy-2,5,7,8- e ame hylch oman-2-ca boxylic acid (T olox),
67
2,2’-azobis (2-me hylp opionamidine) dihyd ochlo ide (AAPH) and 2,2’-diphenyl-1-
68
pic ylhyd azyl (DPPH) we e pu chased om Sigma-Ald ich (S einheim, Ge many).
69
2.2. Samples
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Fo he op imisa ion o he ex ac ion p ocess, we used s awbe ies (F aga ia ananassa
71
a . cama osa) acqui ed a he ma ke . The ui was c ushed in ou labo a o y,
72
dis ibu ed in o ambe glass lasks and ozen a -20º C.
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Fo he p oduc ion o he inega s, we employed h ee di e en ba ches o s awbe ies
74
(F aga ia ananassa a . cama osa) om he Huel a a ea (Spain), co esponding o
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4
h ee ha es s: 2008, 2009 and 2010. The p oduc ion p ocesses we e pe o med in he
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labo a o ies o he Dep o Biochemis y and Bio echnology, Facul y o Oenology, Uni
77
Ro i a i Vi gili (Ta agona). In 2008 and 2009, he subs a e employed we e pu ees
78
p epa ed in he labo a o y using a bea e . In 2010, we used a comme cial pu ee p o ided
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by he Hudisa Company (Huel a). Sulphu dioxide (60 mg/L), suc ose and wo ypes o
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pec oly ic enzymes (Depec il ex a-ga de FCE® and Depec il cla i ica ion® om Ma in
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Viala e Oenologie, Epe nay, F ance), bo h a a concen a ion o 15 mg/L, we e added
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o he pu ee. A e his poin , he p ocedu es we e sligh ly di e en in each ha es .
83
2008 ha es
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One po ion o he s awbe y pu ee was p essed o s udy he e ec o wo ypes o
85
s a ing subs a es (semi-solid and liquid) (Table 1). Six glass con aine s we e illed
86
wi h 6 L o ui subs a e ( ou pu ees and wo liquids). Hal o he con aine s o each
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ype o subs a e we e inocula ed wi h he yeas Saccha omyces ce e isiae QA23 a a
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concen a ion o 2x106 cells/mL, and spon aneous alcoholic e men a ion was allowed
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o occu in he o he hal . All wines we e spon aneously ace i ied keeping i in he same
90
con aine s. Two inal ea men s we e es ed in inega s: pas eu iza ion o
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cen i uga ion. The a e age ace ic deg ees in he 2008 s awbe y inega s we e 4.8.
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2009 ha es
93
Fo he inega p oduc ion in 2009, eigh glass essels we e illed wi h 6 L o
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s awbe y pu ee each. Hal o hese essels we e inocula ed wi h he yeas s ain
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Saccha omyces ce e isiae RP1, isola ed du ing he 2008 spon aneous alcoholic
96
e men a ion, and spon aneous alcoholic e men a ion was allowed o occu in he o he
97
hal . All o he wines ob ained om he inocula ed alcoholic e men a ion we e mixed
98
and dispensed in h ee di e en ypes o con aine s: a glass essel and oak o che y
99
wood ba els. Samples we e hen inocula ed wi h a s ain o ace ic acid bac e ia isola ed
100

5
om he 2008 ace i ica ion. Wines om he spon aneous alcoholic e men a ion we e
101
p ocessed in he same way and le o ace i y spon aneously. The inega s ob ained
102
we e pas eu ised. Inocula ed inega s om he 2009 ha es eached an ace ic deg ee o
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5.5 (glass con aine ), 6.6 (oak ba el) and 6.3 (che y ba el).
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A po ion o he pu ee om he 2009 s awbe ies was concen a ed by hea ing in a
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wa e ba h a 80ºC du ing 10 hou s, o es ano he me hod o inc easing he suga
106
con en ; he esul ing p oduc was a cooked mus (Table 1). The suc ose inal
107
concen a ion was 140 g/L. One li e o his subs a e was e men ed by a spon aneous
108
p ocess and one li e was inocula ed wi h he RP1 s ain o yeas . The inocula ed wines
109
(IWs) we e ace i ied wi h he same ace ic acid bac e ia isola ed in 2008, and he
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spon aneous wines (SWs) we e le o ace i y spon aneously.
111
2010 ha es
112
In his ha es , he pec oly ic enzymes added we e Rohapec ® (12 mg/hL) and he pH
113
was adjus ed o 3.5 wi h 2 g/L CaCO3. In his case, 45 L o pu ee we e e men ed in a
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s ainless s eel con aine on a semi-pilo scale, a e inocula ion wi h S. ce e isiae RP1.
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The ace ous e men a ion was pe o med in a che y wood ba el. The inega had an
116
ace ic deg ee o 6.3.
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All inega s om 2009 and 2010 ha es we e pas eu ized as inal ea men .
118
Fo y-one samples, aken h oughou hese p oduc ion p ocesses, we e analysed. The
119
codes and cha ac e is ics o he samples a e shown in Table 1. In addi ion, i e
120
comme cial inega s we e also analysed o ca y ou compa a i e s udies: Ace o
121
Balsamico, ed wine and whi e wine inega s, apple inega and she y inega .
122
2.3. Sample-ex ac ion p ocedu e
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The consis ency o he samples (pu ees, wines and inega s) made i necessa y o
124
es ablish an ex ac ion sys em p io o analysis. The me hod employed was based on he
125
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ex ac ion p ocedu es designed and op imised p e iously by Ubeda, Hidalgo, To ija,
126
Mas, T oncoso & Mo ales (2011a). Twen y g ams o sample we e mixed in a beake
127
wi h 40 ml o ex ac o 10 min while shaking a 800 pm. The sample was hen
128
subjec ed o ul asonica ion ollowed by a cen i uga ion a 4000 pm o 15 min. The
129
supe na an was eco e ed, and he pelle was e-ex ac ed wi h 40 ml o sol en
130
ollowing he same p ocedu e. Bo h ex ac s we e subsequen ly mixed, and he o ganic
131
sol en was emo ed unde acuum. Finally, he ex ac was il e ed, and MilliQ wa e
132
was added o a inal olume o 15 ml. E e y ex ac ion was pe o med in duplica e. We
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es ed di e en condi ion o ge he maximum alues o AA, TPI and TA as well as
134
economy o sol en used and ime. Thus, he pa ame e s s udied o selec he bes
135
ex ac ion condi ions we e: ype o sol en (ace one, me hanol o e hanol), pe cen age
136
o sol en (80% o 100%) and ul asonic ex ac ion ime (15, 25, 35 o 50 min).
137
2.4. Assays and Me hods
138
2.4.1. ORAC-FL assay
139
The Oxygen Radical Abso bance Capaci y assay (ORAC-FL) was pe o med in a Black
140
96-well mic opla e, ollowing he p ocedu e desc ibed in Ubeda e al. (2011a). This
141
assay was conduc ed in a Mul i-de ec ion pla e eade (Syne gy HT, Ve mon , USA)
142
loca ed a he Cen e o Resea ch, Technology and Inno a ion a he Uni e si y o
143
Se ille (CITIUS). All eac ion assays we e pe o med in iplica e. Resul s we e
144
exp essed as µmol T olox equi alen s (TE)/kg o sample.
145
2.4.2. DPPH adical sca enging assay
146
To de e mine he adical sca enging capaci y, he DPPH assay desc ibed by B and-
147
Williams, Cu elie , & Be se (1995) was used. Fo his es , we used an UV/Vis
148
spec opho ome e U-2800 Digilab coupled o a Pel ie hemos a ic sys em (Hi achi,
149
7
Tokyo, Japan). Resul s we e exp essed as µmol T olox equi alen s (TE)/kg o sample.
150
The assays we e pe o med in iplica e.
151
2.4.3. To al Phenols Index
152
This pa ame e was de e mined in iplica e, using he Folin-Ciocal eu me hod
153
ollowing he p ocedu e desc ibed in Wa e house (2001). Resul s we e exp essed as mg
154
gallic acid/L.
155
2.4.4. To al monome ic an hocyanins
156
The de e mina ion o o al monome ic an hocyanin con en (TA) was measu ed
157
ollowing he pH-di e en ial me hod desc ibed in Gius i & W ols ad (2001). TA was
158
exp essed as pela gonidin-3-glucoside (Plg-3-glu), which is he majo an hocyanin in
159
s awbe y ui wi h a λ is-max a 510 nm (Swain, 1965). Two bu e s we e p epa ed:
160
po assium chlo ide bu e pH=1 (0.025 M), and sodium ace a e bu e pH=4.5 (0.4 M).
161
We measu ed he abso bance a 510 and 700 nm agains a cu e e illed wi h dis illed
162
wa e as a blank.
163
We hen calcula ed he abso bance o he dilu ed sample (A) as ollows:
164
A= (A 510 -A 700) pH 1.0 - (A 510 -A 700) pH 4.5
165
The monome ic an hocyanin concen a ion in he o iginal sample was calcula ed using
166
he ollowing o mula:
167
TA[Plg-3-glu (mg/L)] = (A × MW × DF × 1000) / (ε×1)
168
Whe e
169
A = Sample abso bance
170
MW= Molecula weigh o Plg-3-glu (487.5)
171
DF= Dilu ion ac o
172
ε= Abso p ion coe icien o Plg-3-glu (17330)
173
8
The esul s we e exp essed as mg Plg-3-glu/kg o sample.
174
2.5. S a is ical analysis
175
All s a is ical analysis was pe o med using he S a is ica e sion 7.0 so wa e package
176
(S a so , Tulsa, USA).
177
3. Resul s and discussion
178
3.1. Selec ion o he bes ex ac ion condi ions
179
Se e al ac o s, such as sol en composi ion, ime o ex ac ion, empe a u e, pH, solid-
180
o-liquid a io and pa icle size, may signi ican ly in luence solid-liquid ex ac ions
181
(Azizah, A. H., Ruslawa i, N. M. N., & Tee, T. S, 1999; Pinelo, M., Del Fabb o, P.,
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Manzocco, L., Nunez, M J., & Nicoli, M. C, 2005). In ou case, he pa ame e s ha
183
we e e alua ed o de e mine he bes ex ac ion condi ions we e he ype o sol en , he
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sol en -wa e a io and ul asonica ion ime. The c i e ia used o selec he ex ac ion
185
pa ame e s we e he maximum alues o an ioxidan ac i i y, o al phenols,
186
an hocyanins and ime and sol en sa ings.
187
The ype o sol en is one o he mos in luen ial a iables in he ex ac ion p ocess. We
188
es ed ace one, e hanol and me hanol. The ex ac ion wi h me hanol ga e he wo s
189
esul s in all he assays. As shown in Figu e 1, ace one yielded he highes alues o
190
DPPH (8327 µmol T olox equi alen s (TE)/kg) and TPI (2090 gallic ac. mg/kg), wi h
191
signi ican di e ences in his las pa ame e . Howe e , we ob ained he bes esul s o
192
he ORAC assay (24329 µmol TE/kg) and o he TA de e mina ion (26.78 mg Plg-3-
193
glu/kg) using e hanol, bu no signi ican di e ences we e ound be ween hese alues
194
and hose wi h ace one (26.30 mg Plg-3-glu/kg). Hen íquez, C., Ca asco-Pozo, C.,
195
Gomez, M., B unse , O., & Speisky, H, (2008) epo ed ha he an ioxidan ac i i y o
196
s awbe y ex ac s ob ained wi h ace one/wa e was highe han ha wi h e hanol/wa e
197
and aqueous ex ac s. Taking in o accoun his and o he s udies (Ga cia-Vigue a, C.,
198
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Bosso, A., & Guai a, M. (2008). S udy o some ac o s in ol ed in e hanal p oduc ion
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911−917.
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B and-Williams, W., Cu elie , M. E., & Be se , C. (1995). Use o a ee adical me hod
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o e alua e an ioxidan ac i i y. LWT-Food Science and Technology, 28, 25-30.
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Cano-López, M., Pa do-Minguez, F., López-Roca, J. M., & Gómez-Plaza, E. (2006).
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E ec o mic ooxygena ion on an hocyanin and de i ed pigmen con en and ch oma ic
355
cha ac e is ics o ed wines. Ame ican Jou nal o Enology and Vi icul u e, 57, 325–331.
356
Ce ezo, A. B., Tes aye, W., To ija, M. J., Ma eo, E., Ga cia-Pa illa, M. C., &
357
T oncoso, A. M. (2008). The phenolic composi ion o ed wine inega p oduced in
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ba els made om di e en woods. Food Chemis y, 109, 606-615.
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Ce ezo, A. B., Cue as, E., Win e hal e , P., Ga cia-Pa illa, M. C., & T oncoso, A. M.
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(2010a). Isola ion, iden i ica ion, and an ioxidan ac i i y o an hocyanin compounds in
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Cama osa s awbe y. Food Chemis y, 123, 574-582.
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Ce ezo, A. B., Cue as, E., Win e hal e , M., Ga cia-Pa illa, M. C., & T oncoso, A. M.
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(2010b). An hocyanin composi ion in Cabe ne Sau ignon ed wine inega ob ained by
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subme ged ace i ica ion. Food Resea ch In e na ional, 43, 1577-1584.
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Del eil, D., Feuilla , M., Guilloux-Bena ie , M., Sapis, J. C. (2000). Los inos blancos
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Ga cia-Vigue a, C., Za illa, P., & Tomás-Ba be án, F. A. (1998). The use o ace one as
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an ex ac ion sol en o an hocyanins om s awbe y ui . Phy ochemical Analysis, 9,
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274-277.
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Gius i M. M., & W ols ad, R. E. (2001). Cha ac e iza ion and measu emen o
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an hocyanins by UV–Visible spec oscopy. In R. E. W ols ad, T. E. Ac ee, E. A.
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Häkkinen, S. H., Heinonen, I. M., Kä enlampi, S. O., Mykkänen, H. M., Ruuskanen, J.,
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& Tö önen, A. R. (1999). Sc eening o selec ed la onoids and phenolic acids in 19
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be ies. Food Resea ch In e na ional, 32, 345–353.
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Hannum, S. M. (2004). Po en ial impac o s awbe ies on human heal h: a e iew
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o he science. C i ical Re iews in Food Science and Nu i ion, 44, 1-17.
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Hen íquez, C., Ca asco-Pozo, C., Gomez, M., B unse , O., & Speisky, H. (2008).
380
Slow and as - eac ing an ioxidan s om be ies: hei e alua ion h ough he
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FRAP (Fe ic Reducing An ioxidan Powe ) assay. Ac a Ho icul u ae, 777, 531-
382
536.
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Klopo ek, Y., O o, K., & Boehm, V. (2005). P ocessing s awbe ies o di e en
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p oduc s al e s con en s o i amin C, o al phenolics, o al an hocyanins, and
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an ioxidan capaci y. Jou nal o Ag icul u al and Food Chemis y, 53, 5640-5646.
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Lee, J., & W ols ad, R. E. (2004). Ex ac ion o an hocyanins and polyphenolics om
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bluebe y-p ocessing was e. Jou nal o Food Science, 69, 564-573.
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Mää ä-Riihinen, K. R., Kamal-Eldin, A., & Tö önen, A. R. (2004). Iden i ica ion and
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quan i ica ion o phenolic compounds in be ies o F aga ia and Rubus species ( amily
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Rosaceae). Jou nal o Ag icul u al and Food Chemis y, 52, 6178-6187.
391
Meye , A. S. (2002). Enhanced ex ac ion o an ioxidan phenols om wine and
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juice p ess esidues ia enzyma ic polysaccha ide hyd olysis. F ui P ocessing, 12,
393
29-33.
394
Meye s, K. J., Wa kins, C. B., P i s, M. P., & Liu, R. H. (2003). An ioxidan and
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an ip oli e a i e ac i i ies o s awbe ies. Jou nal o Ag icul u al and Food
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Chemis y, 51, 6887-6892.
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Mo a a, A., Gomez-Co do es, M. C., Colomo, B., & Sua ez, J. A. (2005). Cell wall
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an hocyanin adso p ion by di e en Saccha omyces s ains du ing he e men a ion o
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Vi is ini e a L. c G aciano g apes. Eu opean Food Resea ch and
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Technology, 220, 341-346.
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Olsson, M. E., Ande sson, C. S., O edsson, S., Be glund, R. H., & Gus a sson, K.
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(2006). An ioxidan le els and inhibi ion o cance cell p oli e a ion in i o by
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ex ac s om o ganically and con en ionally cul i a ed s awbe ies. Jou nal o
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Ag icul u al and Food Chemis y, 54, 1248-1255.
405
Pinelo, M., Del Fabb o, P., Manzocco, L., Nunez, M J., & Nicoli, M. C. (2005).
406
Op imiza ion o con inuous phenol ex ac ion om Vi is ini e a byp oduc s. Food
407
Chemis y, 92, 109-117.
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See am, N. P., Lee, R., Scheulle , H. S., & Hebe , D. (2006). Iden i ica ion o phenolic
409
compounds in s awbe ies by liquid ch oma og aphy elec osp ay ioniza ion mass
410
spec oscopy. Food Chemis y, 97, 1-11.
411
Shau-mei, A., & Chang R. C. (2009). Taiwan ui inega . In L. Solie i, & P. Giudici
412
(Eds.), Vinega s o he Wo ld (pp. 223-242). Milan: Spinge -Ve lag I alia.
413
Solie i, L., & Giudici, P. (2009). Vinega s o he wo ld. In L. Solie i, & P. Giudici
414
(Eds.), Vinega s o he Wo ld (pp. 1-16). Milan: Spinge -Ve lag I alia.
415
Swain, T. (1965) Analy ical me hods o la onoids. In T. W. Goodwin (Ed.),
416
Chemis y and Biochemis y o Plan Pigmen s (pp. 543-544). London: Academic P ess.
417
Ubeda, C., Hidalgo, C., To ija, M. J., Mas, A., T oncoso, A. M., Mo ales, M. L.
418
(2011a). E alua ion o an ioxidan ac i i y and o al phenols index in pe simmon
419
inega s p oduced by di e en p ocesses. LWT-Food Science and Technology, 44,
420
1591-1596.
421
18
Ubeda, C., Callejón, R. M., Hidalgo, C., To ija, M. J., Mas, A., T oncoso, A. M.,
422
Mo ales, M. L. (2011b). De e mina ion o majo ola ile compounds du ing he
423
p oduc ion o ui inega s by s a ic headspace gas ch oma og aphy–mass spec ome y
424
me hod. Food Resea ch In e na ional 44, 259–268.
425
Ve beys , L., Oey, I., Van de Plancken, I., Hend ickx, M., & Van Loey, A. (2010).
426
Kine ic s udy on he he mal and p essu e deg ada ion o an hocyanins in s awbe ies.
427
Food Chemis y, 123, 269-274.
428
Wol e, K. L., Kang, X., He, X., Dong, M., Zhang, Q., & Liu, R. H. (2008). Cellula
429
an ioxidan ac i i y o common ui s. Jou nal o Ag icul u al and Food Chemis y, 56,
430
8418-8426.
431
Xu, Q., Tao, W., & Ao, Z. (2007). An ioxidan ac i i y o inega melanoidins. Food
432
Chemis y, 102, 841-849.
433
434
435
436
437
438
439
440
441
442
443
444
445
446
19
447
448
Figu e cap ions
449
Figu e 1. ORAC, DPPH (le axis) and TPI ( igh axis) alues o he di e en
450
ex ac ion sol en s es ed in s awbe ies acqui ed a he ma ke . The ba s in he same
451
assay wi h di e en le e s show signi ican di e ences (p<0.05) (ORAC assay: a, b, c;
452
IPT: A, B, C; DPPH es : α, β, γ).
453
Figu e 2. E ec o sol en pe cen ages. a) ORAC and DPPH alues. b) TPI and
454
TA alues o s awbe ies acqui ed a he ma ke . The ba s in he same assay wi h
455
di e en le e s show signi ican di e ences (p<0.05) (ORAC and TPI assays: a, b;
456
DPPH and TA es s: A, B).
457
Figu e 3. E ec o di e en ul asonica ion imes a) ORAC and DPPH
458
alues. b) TPI and TA alues o s awbe ies acqui ed a he ma ke . The
459
ma ke s in he same assay wi h di e en le e s show signi ican di e ences (p<0.05)
460
(ORAC and TPI assays: a, b, c; DPPH and TA es s: A, B, C).
461
Figu e 4. Compa ison o ORAC, DPPH (le axis) and TPI ( igh axis) alues o
462
s awbe y inega s wi h comme cial a ie ies. Sample codes: F9MCV (mean alue o
463
all inega s om cooked mus ), F9V (mean alue o all inega s om 2009 ha es )
464
and F8V (mean alue o all inega s om 2008 ha es ).
465
466
467
468
469
470
471
20
Figu e 1.
472
473
474
475
476
477
478
479
480
481
482
483
484
485
486
Figu e 2.
487
488
489
Figu e 3.
490
491
492
493
494
495
496
497
a
b
a
α
ß
α
C
B
A
0
5000
10000
15000
20000
25000
30000
Ace one Me hanol E hanol
µmol TE/kg R
0
500
1000
1500
2000
2500
mg gallic acid/kg
b
a
B
A
0
5000
10000
15000
20000
25000
80% 100%
µmol TE/Kg
0
1000
2000
3000
4000
5000
6000
7000
8000
9000
µmol TE/Kg
a
b
A
B
0
500
1000
1500
2000
2500
80% 100%
mg gallic acid/Kg
0
5
10
15
20
25
30
mg Plg-3-glu/Kg
a)
b)
a)
b)
b
b
a
a
C
B
A
A
22000
23000
24000
25000
26000
27000
28000
29000
15 25 35 50
µmol TE/Kg
5000
5500
6000
6500
7000
7500
8000
8500
9000
9500
µmol TE/Kg
a
b
c
d
A
B
A
A
2000
2100
2200
2300
2400
2500
2600
2700
15 25 35 50
mg gallic acid/Kg
40
45
50
55
60
65
70
mg Plg-3-glu/Kg
a)
b)

21
Figu e 4.
498
499
500
501
502
503
504
505
506
507
508
509
510
511
512
513
514
515
516
517
518
519
520
521
522
523
0
5000
10000
15000
20000
25000
30000
35000
40000
45000
Balsamic
F9MCV
F10VI
F9V
F8V
Apple
She y wine
Red wine
Whi e wine
µmol TE/kg
0
500
1000
1500
2000
2500
3000
mg gallic acid/kg
32
Table 1. Samples desc ip ion.
Ha es
T ea men
Pu ee
Sample
T ea men
Sample
subs a e
Alcoholic
e men a ion
( ime)
Wine Sample
Ace i ica ion
( ime)
T ea men o
Recipien
Vinega sample
2008
C ushed
F8P1
SO2
Pec oly ic
enzymes
Suc ose (50 g/L)
F8P2
Inocula ed
(4 days)
F8WI1- F8WI4
Spon aneous
(2 mon hs)
Cen i uga ion
F8VIC1-F8SVIC2
Pas eu iza ion
F8SVIP1-F8SVIP2
Spon aneous
(5 days)
F8WE1- F8WE4
Cen i uga ion
F8SVEC1-F8SVEC2
Pas eu iza ion
F8SVEP1-F8SVEP2
-
F8P2
P essing
F8L
Inocula ed
(4 days)
F8LWI
-
-
-
Spon aneous
(5 days)
F8LWE
2009
C ushed
F9P1
SO2
Pec oly ic
enzymes
Suc ose (75 g/L)
F9P2
Inocula ed
(5 days)
F9WI1- F9WI4
Inocula ed
(2 mon hs)
glass essel
F9SVIG
oak ba el
F9SVIO
che y ba el
F9SVIX
Spon aneous
(8 days)
F9WE1- F9WE4
Spon aneous
(2 mon hs)
glass essel
-
oak ba el
-
che y ba el
-
Hea ing
Concen a ed
F9MC
Inocula ed
(7 days)
F9MCWI1-F9MCWI2
Inocula ed
(5 mon hs)
glass essel
F9MCVI1-F9MCVI2
Spon aneous
(7 days)
F9MCWE1-F9MCWE2
Spon aneous
(2.5 mon hs)
glass essel
F9MCVE1-F9MCVE2
2010
C ushed
F10P1
SO2
Pec oly ic
enzymes
Suc ose (65 g/L)
CaCO3
F10P2
Inocula ed
(4 days)
F10WI
Inocula ed
(1.5 mon hs)
che y ba el
F10VI
33
Table 2. Changes in 2008 samples on ORAC, DPPH, TPI and TA du ing s awbe y inega p oduc ion (a e age±s anda d de ia ion).
1
Samples
ORAC
(μmol TE/kg)
DPPH
(μmol TE/kg)
TPI
(mg gallic acid /kg)
TA
(mg plg-3-glu/kg)
Subs a es
F8P1
21792 ± 221
8327 ± 99
2090 ± 10
26.3 ± 0.8
F8P2
26714 ± 910a
10116 ± 88a
2298 ± 0a
69 ± 0a
F8L
20642 ± 111b
5907 ± 516b
1615 ± 33b
43 ± 0b
Wines
F8LWE
12757 ± 267b,c
2837 ± 59b,c
868 ± 29b,c
12.2 ± 0.2b
F8LWI
13497 ± 227b,c
2898 ± 129b,c
858 ± 13b,c
17.9 ± 0.2b,d
F8SWE1
25314 ± 650
8200 ± 58b
1907 ± 26
13.1 ± 0.7b
F8SWE2
24696 ± 70
7879 ± 70b
1773 ± 32
12.9 ± 0.6b
F8SWE3
25458 ± 403
7689 ± 82b
1757 ± 45
12.4 ± 0.7b
F8SWI1
27987 ± 1227b
7241 ± 35b,d
1670 ± 9b,d
16 ± 0b,d
F8SWI2
25451 ± 429b
8004 ± 35b,d
1584 ± 19b,d
18.0 ± 0.3b,d
F8SWI3
23745 ± 15b
6515 ± 67b,d
1548 ± 6b,d
17.3 ± 0.6d
Vinega s
F8SVE1C
9202 ± 390b
3256 ± 205b
769 ± 13b
0.4 ± 0.0b
F8SVE1P
9849 ± 413b
3368 ± 352b
774 ± 23b
0.5 ± 0.1b
F8SVE2C
9215 ± 338b
3210 ± 129b
781 ± 0b
1.1 ± 0.2b
F8SVE2P
10869 ± 190b
3252 ± 234b
683 ± 10b
0.6 ± 0.0b
F8SVI1C
10139 ± 341b,e
3227 ± 117b
751 ± 16b
1.3 ± 0.0b
F8SVI1P
11611 ± 89b,e
3388 ± 64b
744 ± 6b
0.9 ± 0.1b
F8SVI2C
11054 ± 40b,e
3260 ± 246b
694 ± 16b
0.8 ± 0.1b
F8SVI2P
11082 ± 86b,e
3380 ± 76b
712 ± 9b
1 ± 0b
34
Sample codes a e loca ed in Table 1.
a Signi ican di e ences (p<0.05) wi h espec o he ini ial ui pu ee (ANOVA).
b Signi ican di e ences (p<0.05) wi h espec o he sample om which was p oduced (ANOVA).
c Signi ican di e ences (p<0.05) wi h espec o semisolid wines ob ained wi h simila alcoholic
p ocess (spon aneous o inocula ed) (ANOVA).
d Signi ican di e ences (p<0.05) wi h espec o spon aneous p ocess (ANOVA).
e Signi ican di e ences (p<0.05) wi h espec o he inega s ob ained om spon aneous wines (ANOVA).
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17