1. In oduc ion
In ecen yea s, many pape s ha e paid a en ion o he
bioac i e compounds, pa icula ly o he an ioxidan ac i i y
o polyphenolic compounds in ood and be e ages, due
o hei posi i e e ec on he human body. As consume s
ha e become mo e conscious o he heal h bene i s
o phenolic compounds and hei an ioxidan ac i i ies
ia he con en ional media, he be e age indus ies
ha e ecognized new ma ke ing oppo uni ies o hei
p oduc s. The e o e, he phenolic compounds and hei
an ioxidan capaci y in oods and be e ages become an
impo an quali y pa ame e , especially in niche ma ke s
conce ned wi h heal h bene i s.
Wine is a widely consumed be e age in he wo ld,
wi h housands o yea s o adi ion. I is an excellen
sou ce o a ious classes o polyphenols. The
phenolic compounds a e esponsible o he senso y
cha ac e is ics, pa icula ly colo , as ingency, bi e ness
and a oma [1,2]. The phenolic compounds in ed wine
exhibi a boa d spec um o bene icial pha macological
p ope ies, belie ed o be ela ed o hei an ioxida i e
p ope ies. An i-a he ogenic, an i- umou , an i-ulce , and
an i-in lamma o y ac i i ies ha e all been demons a ed
by he consump ion o ed wine and ed wine phenolic
compounds [3-8]. As one o he winemaking p ocedu es,
he phenolic compounds o he wine g ape a e one o
he mos impo an aspec s ha de e mine wine quali y.
A la ge numbe o published pape s ha e ocused on he
essen ial con ibu ions o phenolic compounds p o iles o
wine quali y and senso y p ope ies [1,2].
The phenolic p o iles in wine depend on he phenolic
compounds p esen in he g apes, he ex ac ion
pa ame e s, winemaking echnologies as well as
e men a ion empe a u e, yeas s ain, p ocessing
enzymes, cap managemen , and alcohol concen a ion
[9-11]. On he o he hand, he phenolic compounds
o g apes a e a ec ed by many ac o s such as ag o
Cen al Eu opean Jou nal o Chemis y
* E-mail: [email p o ec ed]
1Depa men o Biochemis y and Food Analysis,
Tomas Ba a Uni e si y in Zlín, CZ-762 72 Zlín, Czech Republic
2Ins i u e o Chemis y, Depa men o Analy ical Chemis y,
Masa yk Uni e si y in B no, CZ-62500 B no, Czech Republic
Badam se seg Soyollkham1, Pa el Valášek1, Mi osla Fiše a1, Vlas imil Fic1,
Vlas imil Kubáň1,2, Ignác Hoza1
To al polyphenolic compounds con en s (TPC),
o al an ioxidan ac i i ies (TAA) and HPLC
de e mina ion o indi idual polyphenolic
compounds in selec ed Mo a ian
and Aus ian wines
Resea ch A icle
Abs ac :
© Ve si a Sp. z o.o.
Recei ed 9 No embe 2010; Accep ed 23 Ma ch 2011
Keywo ds: Phenolic compounds • Wine • To al an ioxidan ac i i y • To al phenolic con en • HPLC/UV-VIS
Wine samples (G üne Vel line (GV) and Zweigel (ZW) om ou di e en geog aphical egions o Aus ia and Czech Republic) we e
analyzed o de e mine hei o al phenolic con en (TPC) by applying he Folin-Ciocal eau me hod, o al an ioxidan ac i i y (TAA)
by FRAP ( e ic educing an ioxidan powe ) and DPPH (1,1-diphenyl-2-pic yl-hyd azyl) assays, and o iden i y and quan i y ele en
phenolic compounds using a HPLC/UV-VIS me hod.
Cen . Eu . J. Chem. • 9(4) • 2011 • 677-687
DOI: 10.2478/s11532-011-0045-3
677
To al polyphenolic compounds con en s (TPC), o al an ioxidan
ac i i ies (TAA) and HPLC de e mina ion o indi idual polyphenolic
compounds in selec ed Mo a ian and Aus ian wines
echnical p ocesses, gene ic a ia ion, ma u i y, clima ic
and geog aphical condi ions [12-14]. O he ac o s
ha in luence he ex en o phenolic ex ac ion a e he
molecula weigh , size and ype o phenolic molecules,
he su ace a ea, he concen a ion g adien , o he
empe a u e ea men s including g ape and mus
eezing and he mo- ini ica ion, and ac o s ha a ec
cell pe meabili y, such as pec oly ic enzyme selec ion
[15]. Also, he en i onmen al condi ion ( empe a u e,
annual p ecipi a ion le els, al i ude and geochemical
cha ac e is ics) can a ec he g apes ma u a ion
and consequen ly he concen a ion o hei phenolic
compounds.
Many pape s dealing wi h phenolic compounds o
wine and g apes and hei o al an ioxidan capaci y
ha e been published. Howe e , li le a en ion has been
paid o compa ing he phenolic compounds o wine
g apes om di e en o igins in Mo a ian wine, as well
as compa ing he phenolic con en s and an ioxidan
ac i i ies o phenolic compounds. Fla onoids, phenolic
acids, la onols and ans- es e a ol and o he g oups
o compounds could be key agen s o he an ioxidan
ac ion on he human me abolism pa hway, he eason
why we a e quali ying he wines om a nu i ional poin
o iew.
This s udy de e mines he o al con en o phenolics,
iden i ies and quan i ies indi idual phenolic compounds
and de e mines he o al an ioxidan ac i i y in wine
samples collec ed om ou di e en geog aphical
egions o Aus ia and Czech Republic ( wo wine ies
in Aus ia – Poysdo and G oss ieden hal; and wo
wine ies in he Czech Republic - Velké Bílo ice and
Velké Hos ě ádky - Bošo ice). The s udy assesses he
in luence o di e en geog aphical condi ions on he
phenolic composi ion and e alua es he ela ionship
be ween an ioxidan po en ial and he phenolic con en
o Mo a ian and Aus ian wine.
2. Expe imen al P ocedu e
2.1. Ins umen a ion
A single beam lash scan diode a ay spec opho ome e
co e ing 330-800 nm Bioch om Lib a S6 (Bioch om
L d, Camb idge, UK) was used o spec opho ome ic
assays. The Ul iMa e® 3000 HPLC sys em consis ed o
Ul iMa e 3000 RS pump, Ul iMa e 3000 RS au osample ,
Ul iMa e 3000 RS column compa men and Ul iMa e
3000 RS diode a ay de ec o (Dionex Co., Sunny ale,
CA, USA). Ch oma og aphic sepa a ion was ca ied ou
on Supelcosil LC-18-DB column (250×4.6 mm, 5 µm,
Supelco, USA) a 30°C by g adien elu ion wi h a mobile
phase con aining sol en A (5% / aqueous ace oni ile
(ACN) acidi ied wi h 0.35 mL i luo oace ic anhyd ide
(TFAA) and sol en B (50% / aqueous ace oni ile
acidi ied wi h 0.25 mL TFAA). Run ime was 30 min and
he low a e was 1 mL min-1.
2.2. Chemicals
Folin-Ciocal eau eagen , gallic acid, 2,4,6- is-(-2-
py idyl)-s- iazine (TPTZ) and 2,2-diphenyl-1-pic yl-
hyd azyl (DPPH) we e ob ained om Sigma-Ald ich
(S einheim, Ge many). A s anda d solu ion o DPPH
c = 0.20 mol L-1 was p epa ed in me hanol. A wo king
DPPH solu ion was p epa ed a c = 100 µmol L-1
con aining an ace a e bu e o pH 4.3 wi h a a io o 1:2
(DPPH:bu e ). Tannin was ob ained om Me ck KGaA
(Da ms ad , Ge many). Phenolic e e ence s anda ds
including gallic acid, ca echin, anillic acid, ca eic acid,
p-couma ic acid, e ulic acid, sinapic acid, u in, cinnamic
acid, que ce in and es e a ol we e pu chased om
Ald ich (Zwijnd ech , Belgium). Lab-Scan ace oni ile
(ACN) was ob ained om POCH A.S. (Gliwice, Poland).
O he chemicals and eagen s o eagen g ade pu i y
we e pu chased om Pen a, Ch udim and/o Lachema,
B no, (bo h Czech Republic). All solu ions we e p epa ed
wi h deionised (DI) wa e (Aquaosmo ic, Tišno , Czech
Republic).
2.3. Sampling
A o al o 32 wine samples including 16 whi e and 16 ed
wines (all wines om he 2009 in age) we e collec ed
be o e bo ling in Vinopol L d. (Velké Bílo ice, Czech
Republic). Wines we e made om G üne Vel line (GV)
and Zweigel (ZW) wine g apes g own in ou di e en
geog aphical egions o Aus ia and Czech Republic; wo
wine ies in Aus ia (Poysdo and G oss ieden hal) and
wo wine ies in he Czech Republic (Velké Bílo ice and
Velké Hos ě ádky - Bošo ice). Fou locali ies in mu ual
dis ances (mo e han 40 km) we e selec ed o sampling
o minimize he isks posed by wea he condi ions.
The Aus ian ineya ds o he wine ies we e loca ed
in wo dis ic s. Poysdo is loca ed in Aus ia’s la ges
wine-g owing a ea called “Wein ie el” (16.650 ha).
G oss ieden hal is embedded in he wine-g owing a ea
called “Wag am” (2.800 ha). In he Czech Republic, he
ineya ds a e loca ed in he Mo a ian sub-dis ic called
“Velkopa lo ická“ (5.200 ha). G apes we e ha es ed in
ou epe i ion uni s (see Fig. 1) om each ineya d; wine
samples we e made by each epe i ion g ape samples.
Table 1 shows a lis o analyzed wine samples.
The clima ic condi ions o he wo wine-g owing
a eas in Aus ia show nonsigni ican di e ences.
Poysdo (225 m abo e sea le el) and G oss ieden hal
(277 m abo e sea le el) a e a ec ed by he Pannonian
clima e ha is cha ac e ized by ho d y summe s and
678
B. Soyollkham e al.
cold win e s. The annual a e age empe a u e is 9.1°C
and he annual p ecipi a ion is 480 mm. The geology
o he egion shows ha Poysdo (embedded in he
Vienna basin) and G oss ieden hal (embedded in he
Molasse basin) is cha ac e ized by loess and loess-
clay. The pedology o he ineya ds di e s be ween
he wo wine-g owing a eas; in Poysdo he soil ype
is a i gin soil and in G oss ieden hal i is a b own soil.
In he Czech Republic, he ineya ds in Velké Bílo ice
(185 m abo e sea le el) and Velké Hos ě ádky - Bošo ice
(215 m abo e sea le el) a e a ec ed by he con inen al
clima e wi h an occasional in luence o A lan ic clima e.
The annual a e age empe a u e is 9.4°C and he
annual p ecipi a ion is 510 mm. The geology o he wine-
g owing a ea is cha ac e ized by calca eous clay, ma l
and sands one. Vi gin soil cha ac e izes he soil ype o
he ineya ds .
2.4. Sampling s a egy and wine making
p ocedu es
Sample p ocedu es we e pe o med in acco dance
wi h he ollowing c i e ia in di e en geog aphical
egions o he expe imen al ields: i) 400 g ape ines
pe expe imen al ield, ii) 4 epe i ion uni s, iii)
c op age 6 – 25 yea s, i ) dis ance be ween ows
2 – 2.5 m, ) applica ion o ull mechaniza ion and
iennial dep h-loosene , i) b eak ou o g ape ines
8 – 10 pe in age.
Acco ding o a s anda d winemaking p ocedu e,
he e is no wood con ac in he expe imen al cella o
Vinopol. A e c ushing 100 kg o g apes, diammonium
phospha e, SO2 and Saccha omyces ce e isiae s ain
we e added o each wine . The g ape skin was sepa a ed
om he juice using s anda d p essing p ocedu e.
P essed juice was added o he 50-L glass bo les and
e men ed by s anda d p ocedu e a 8°C, and he cap
was punched down wo imes pe day. Wines we e cold-
s abilized o se e al weeks a 8°C and il e ed using
ce amic il e s.
2.5. Me hods
2.5.1. Folin-Ciocal eau me hod
The TPC was de e mined acco ding o he Folin-
Ciocal eau me hod [16]. B ie ly, a 0.025 mL sample was
mixed wi h 1 mL o he 10- old dilu ed Folin-Ciocal eau
eagen and allowed o s and o 3 min. Then 5 mL o
200 g L-1 sodium ca bona e (Na2CO3) was added, and
a inal olume was made up o 50 mL wi h DI wa e .
Each sample was measu ed spec opho ome ically
a 765 nm a e 30 min o s anding agains he blank.
Fi e-poin calib a ion was s ic ly linea (R2>0.9999)
in he concen a ion ange 0 - 250 mg L-1 wi h annin
as he s anda d. The eg ession equa ion was
A = 0.0018 c + 0.0028. The de e mined alues
we e exp essed as annin equi alen s (TE, mg L-1).
All samples we e analyzed as iplica es. S anda ds and
samples gleaned highly epea able esul s.
2.5.2. DPPH adical sca enging ac i i y [17,18]
A mix u e o an undilu ed sample (0.1 mL) wi h
a 10 mL wo king DPPH solu ion was measu ed
immedia ely a 515 nm agains a me hanol blank (AC(0)
).
The mix u e was hen incuba ed a oom empe a u e
and in he da k o 30 minu es and was again measu ed
spec opho ome ically a 515 nm (AA( )). The gallic acid
(GA) calib a ion cu e was plo ed as a unc ion o he
pe cen age o he DPPH adical sca enging ac i i y.
The measu emen was compa ed o he s anda d
calib a ion cu e, and he ee adical sca enging
ac i i ies we e exp essed as mic omoles o gallic acid
equi alen s (GAE) pe millili e o sample (µmol mL-1).
The calib a ion cu e was s ic ly linea (A = 855.59,
c = 16.015, R2 = 0.9980, whe e A is abso bance alue,
c is he concen a ion o gallic acid in s anda d solu ions)
in he concen a ion in e al 0.02 – 0.08 µmol mL-1 gallic
acid. The µmol mL-1 inhibi ion o he DPPH adical
caused by a wine sample we e de e mined acco ding
o he ollowing o mula: (AC(0) – AA( ))/AC(0) × 100, whe e
AC(0) is he abso bance o he sample a = 0 min and AA( )
is he abso bance o sample a = 30 min). All samples
we e analyzed as iplica es.
Table 1. Lis o analysed wine samples
Sample
Code*
Type o
wine
Vineya ds
(dis ic ) Wine ies
SGV Whi e Velkopa lo ická Velké Bílo ice
SZW Red
PGV Whi e Velkopa lo ická Velké Hos ě ádky - Bošo ice
PZW Red
OGV Whi e Wein ie el Poysdo
OZW Red
BGV Whi e Wag am G oss ieden hal
BZW Red
* GV – G uene Vel line , ZW – Zweigel , a ia ions 1 – 4 acc. Fig. 1
we e collec ed
Figu e 1. Plan o he expe imen al ields o sampling om
indi idual expe imen al ineya ds in Czech Republic (S, P)
and Aus ia (O, B).
679
To al polyphenolic compounds con en s (TPC), o al an ioxidan
ac i i ies (TAA) and HPLC de e mina ion o indi idual polyphenolic
compounds in selec ed Mo a ian and Aus ian wines
2.5.3. Fe ic ion educing an ioxidan powe (FRAP)
The educing ac i i y o he samples was de e mined by
he FRAP me hod [19]. A 0.1 µmol L-1 s anda d solu ion
o gallic acid (GA) was p epa ed in H2O. The oxidan
in he FRAP assay was p epa ed by mixing 5 mL o
10 mmol L-1 2,4,6- ipy idyl-s- iazine (TPTZ) in wa e ,
50 mL o ace a e bu e pH 3.6, and 5 mL o FeCl3•H2O
(20 mmol L-1). A sample (25 µL) was added o
he 4 mL eagen . Abso bance was measu ed
spec opho ome ically a 593 nm (A0min). Then
he sample solu ion was allowed o s and a oom
empe a u e and in he da k o 10 min and measu ed
again a 593 nm (A10min). The di e ence o abso bances
(ΔA = A10min - A0min) o he eac ion mix u e was
calcula ed and ela ed o ΔA o a Fe(II) s anda d
solu ion. The di e ence in abso bance ΔA was linea ly
p opo ional o he concen a ion o he an ioxidan and
indica ed inc eased educing powe . The measu emen
was compa ed o a calib a ion cu e o he p epa ed
gallic acid solu ion, and hen inal esul s we e exp essed
as mic omoles o gallic acid equi alen s (GAE) pe
millili e o he sample (µmol mL-1). The calib a ion cu e
was s ic ly linea (A = 1.0800 c + 0.0072, R2 = 0.9999,
whe e A is he abso bance alue, c is he concen a ion
o gallic acid in s anda d solu ions) in he concen a ion
in e al 0.02 – 0.1 µmol mL-1 gallic acid. All samples
we e analyzed as iplica es.
2.5.4. HPLC analysis o phenolic composi ion
The indi idual phenolic compounds we e quan i ied
using a HPLC me hod [20] using g adien elu ion wi h
he mobile phase con aining sol en A (5% / aqueous
ACN acidi ied wi h 0.35 mL i luo oace ic anhyd ide
(TFAA) and sol en B (50% / aqueous ACN acidi ied
wi h 0.25 mL TFAA). The UV de ec o was se a 205,
210, 275 and 375 nm. The wine sample was il e ed
using 0.45 µm po e size Nylon memb ane il e 13 mm
(FFNN1345-100, G onus, SMI labHu L d., Maisemo e,
Glouces e shi e, UK) using il e de ices (Millipo e,
Bed o d, MA, USA) be o e injec ing. The injec ion
olume was 20 µL. Indi idual phenolic compounds we e
iden i ied by compa ing e en ion imes and UV spec a
o he co esponding s anda d compounds. Da a we e
quan i ied using he co esponding calib a ion cu es o
he indi idual s anda d compound.
3. Resul s and Discussion
In his s udy, a o al o 32 wine samples including 16
whi e and 16 ed wines, which we e made om he
G üne Vel line and Zweigel g ape a ie ies, we e
selec ed o de e mine he o al phenolic con en s (TPC)
and he o al an ioxidan ac i i y (TAA). G üne Vel line
is a a ie y o whi e wine g ape g own p ima ily in Aus ia
and in he Czech Republic. Zweigel is a ed wine g ape
a ie y ha is he mos widely g own in Aus ia oday.
3.1. To al phenolic con en s
The di e en a ia ions o ed and whi e wine samples
we e es ed o TPC in ou se s o analyses. The TPC
a ied om 218 o 328 mg L-1, a e aging 263 mg L-1,
o he ou whi e wine samples SGV and om 1182 o
1232 mg L-1, a e aging 1216 mg L-1, o he ou ed wine
samples SZW om Velké Bílo ice. The o al phenolic
con en s anged om 268 o 283 mg L-1, a e aging
274 mg L-1 o PGV samples and om 564 o
729 mg L-1, a e aging 651 mg L-1 o ed wine samples
PZW om Velké Hos ě ádky - Bošo ice. Samples PGV-3
and PZW-3 showed a high con en o o al phenolics;
he same as SGV-3 and SZW-3. I can be assumed ha
he high eading o he phenolic con en s o he g ape
samples depends on he loca ion o he ineya d whe e
he g apes g ew, hei shel e om he wind, in ensi y
o sunligh adia ion as well as shaded o non shaded
clus e s and o he ac o s.
In he nex se o analyses, wine samples om
Aus ian egions we e es ed ( ou ed wines and ou
whi e wines). The TPC a ied om 260 o 304 mg L-1,
a e aging 275 mg L-1, o he whi e wine samples OGV
and om 824 o 878 mg L-1, a e aging 846 mg L-1,
o he ed wine samples OZW collec ed in Poysdo .
The co esponding alues om 117 o 210 mg L-1,
a e aging 179 mg L-1, in whi e wine BGV, and om 1068 o
1184 mg L-1, a e aging 1122 mg L-1 TE, in ed wine BZW
om G oss ieden hal we e ob ained. In his se , he
highes TPC o he ed wines was ound in OZW-4 and
o he whi e wines was ound in OGV-3 samples.
Figs. 2 and 3 show (see also Table 2) he TPC in
whi e and ed wine samples. The mean alues o he
o al phenolic con en s in whi e wine samples we e
de e mined in he in e al om 263 o 275 mg L-1 TE,
excep o he BGV (179 mg L-1 TE). Ou alues o whi e
wines we e abou 50% highe compa ed o he esul s
published by Simone i e al. [21] and app oxima ed
he alues published by Komes e al. [22], Sánche -
Mo eno e al. [23], Heinonen e al. [24], Jewell [25] and
S e ana o e al. [26]. The TPC o eigh whi e wines om
Sou h Mo a ia we e de e mined by S a il e al. [27] and
alues o TPC we e app oxima ely 40-60 % lowe han
ou esul s. Fig. 2 shows he an ioxidan ac i i y o ou
di e en a ia ions g own in ou di e en ineya ds,
a o al o 16 whi e wine samples. As shown in his
igu e, he highes TPC we e ound o wine samples O
(275 mg L-1) g own in Wein ie el, Aus ia and
P (274 mg L-1) g own in Velké Hos ě ádky - Bošice,
Czech Republic.
680
B. Soyollkham e al.
The a e age alues o TPC in ed wine samples
anged om 651 (PZW) o 1216 (SZW) mg L-1 TE.
These esul s we e in he ange o he p e iously
summa ized [27-29] da a (824-4059 mg L-1) by C ozie
e al. [28], excep o he PZW samples (564-729 mg L-1).
Mo eo e , TPC in ed wine we e app oxima ely 40-75%
lowe compa ed o he esul s published by selec ed
au ho s [21-28,30,31].
The TPC o 16 ed wine samples we e compa ed
wi h each o he . The esul s a e shown in Fig. 3. The
highes TPC we e de e mined o ed wine samples
S (mean alue o ou a ia ions, 1216 mg L-1) g own
in Velkopa lo ická, he Czech Republic and o ed
wine samples B (mean alue o ou a ia ions,
1122 mg L-1) g own in Wag am, Aus ia. Howe e , he ed
wine samples showed a low con en o o al phenolics
o wine samples O which we e g own in Wein ie el,
Aus ia.
Gene ally, he o al phenolic con en s o whi e wine
samples we e obse ed a ela i ely high le els (117-
328 mg L-1), bu hey we e s ill a leas 5- imes lowe
han he ed wines (564-1216 mg L-1). In he whi e wine
samples, he lowes TCP (117 mg L-1) was de e mined in
wine samples B g own in Wag am, bu in con adic ion,
o ed wine samples he high con en was ound in wine
samples B g own in he same egion. These esul s
indica ed ha he geog aphical o igin, a e age annual
empe a u e, annual le els o p ecipi a ion and pedology
s ongly in luenced he wine quali y and quan i y.
3.2. An ioxidan ac i i y o wines
The o al an ioxidan ac i i y alues o he wine samples,
using he DPPH and FRAP me hods, a e gi en in
Figs. 4 and 5. Wine samples showed high TAA, mainly
hose wi h high le els o o al phenolic con en s.
Acco ding o he FRAP alues de e mined in whi e
wines, he TAA can be quali ied in dec easing o de
(GAE mmol L-1): OGV-3 (0.50) ~ OGV-4 (0.50) > PGV-3
(0.43) ~ SGV 3 (0.43). These esul s indica ed ha
OGV and SGV ha e a highe an ioxidan ac i i y han
PGV and BGV whi e wines. The highes an ioxidan
ac i i y was ound in BGV-2 and BGV-1 ed wines
(3.8-3.5 mmol L-1, i.e., 7 o 11- imes highe han ha
in whi e wines). The lowes alue, less han hal o
he highes alue, was ound in PZW-4 ed wine
(1.4 mmol L-1). The a e age an ioxidan ac i i y alues
in each ineya d de e mined by he FRAP me hod we e
0.43 (SGV), 0.4 (PGV), 0.49 (OGV) and 0.38 (BGV)
in whi e wines, mo eo e , 2.9 (SGV), 1.53 (PGV),
1.73 (OGV) and 3.46 (BGV) mmol L-1 in ed wines,
espec i ely.
Fu he mo e, acco ding o he DPPH a e age alues
de e mined in whi e wines, he o al an ioxidan ac i i y
can be quali ied in his dec easing o de (GAE mmol L-1):
PGV (0.35) > OGV (0.32) > BGV-3 (0.3) > SGV (0.29).
These esul s we e in disag eemen wi h he alues
o he highes an ioxidan ac i i y in whi e wines which
we e de e mined by FRAP me hod. Howe e , esul s o
he highes an ioxidan ac i i y in ed wine was ound in
BGV-2 ed wine (2.01 mmol L-1, i.e., 5- o 8- imes highe
han ha in whi e wines). Also,, he lowes alue, less
han hal o he highes alue, was ound in he PZW and
OZW ed wine samples (0.67 and 0.68 mmol L-1). These
esul s we e in ag eemen wi h he alues de e mined by
he DPPH me hod. The mean alues o he an ioxidan
ac i i ies in each ineya d de e mined by he DPPH
me hod we e 0.43 (SGV), 0.4 (PGV), 0.49 (OGV)
and 0.38 (BGV) mmol L-1 in whi e wines, mo eo e ,
2.9 (SGV), 1.53 (PGV), 1.73 (OGV) and 3.46 (BGV)
mmol L-1 in ed wines, espec i ely.
I was di icul o compa e ou alues o he TAA wi h
he li e a u e da a. The majo i y o au ho s ha e used
a ious me hods such as inhibi ion o lipid oxida ion, he
ORAC me hod, he ABTS me hod, he DPPH me hod
wi h he e alua ion o EC50 ( he sample concen a ion
necessa y o educe he emaining DPPH by 50%).
On he o he hand, i is possible o pa ially e alua e,
some o he alues de e mined by he FRAP, DPPH
me hods using he T olox s anda d, e.g. S a il e al.
[27]. Fo he DPPH me hod, alues we e de e mined
Figu e 3. To al phenolic con en s in 16 ed wines om indi idual
expe imen al ineya ds in Czech Republic (S, P)
and Aus ia (O, B). SDs alues see Table 2.
Figu e 2. To al phenolic con en s in 16 whi e wine samples om
indi idual expe imen al ineya ds in Czech Republic
(S, P) and Aus ia (O, B). SDs alues see Table 2.
681

To al polyphenolic compounds con en s (TPC), o al an ioxidan
ac i i ies (TAA) and HPLC de e mina ion o indi idual polyphenolic
compounds in selec ed Mo a ian and Aus ian wines
Table 2.
Phenolic
compounds
Sample
No
Whi e wines Red wines
SGV PGV OGV BGV SZW PZW OZW BZW
15.70 ± 0.00 5.79 ± 0.01 5.53 ± 0.01 5.71 ± 0.02 14.5 ± 0.03 8.93 ± 0.01 14.2 ± 0.02 16.8 ± 0.08
25.66 ± 0.00 5.67 ± 0.31 6.08 ± 0.01 6.01 ± 0.02 14.7 ± 0.01 9.46 ± 0.02 12.9 ± 0.02 17.4 ± 0.04
Gallic acid 35.71 ± 0.00 5.80 ± 0.05 5.83 ± 0.01 5.44 ± 0.02 14.2 ± 0.05 9.65 ± 0.08 14.4 ± 0.02 16.8 ± 0.04
45.04 ± 0.31 5.71 ± 0.01 6.19 ±0.01 5.36 ± 1.00 14.2 ± 0.05 9.23 ± 0.01 14.1 ± 0.02 16.9 ± 0.02
A e age 5.52 ± 0.32 5.74 ± 0.06 5.90 ± 0.29 5.63 ± 0.29 14.4 ± 0.23 9.31 ± 0.31 13.9 ± 0.68 16.9 ± 0.30
15.01 ± 0.03 9.90 ± 0.0 4.63 ± 0.01 6.46 ± 0.07 23.5 ± 14.85 23.5 ± 0.03 25.6 ± 0.04 20.2 ± 0.28
28.08 ± 0.02 10.57 ± 0.01 4.21 ± 0.01 8.38 ± 0.05 38.7 ± 0.01 13.8 ± 0.02 25.0 ± 0.01 22.8 ± 0.05
Ca echin 310.11 ± 0.07 11.44 ± 0.76 6.21 ± 2.23 7.00 ± 0.03 29.7 ± 0.46 24.9 ± 0.47 23.9 ± 0.04 25.1±0.82
46.77 ± 0.02 10.69 ± 0.09 6.79 ± 0.03 6.63 ± 0.04 29.7 ± 0.46 24.1 ± 0.02 24.8 ± 0.03 24.7 ± 0.04
A e age 7.49 ± 2.15 10.65 ± 0.63 5.46 ± 1.23 7.11 ± 0.87 30.4 ± 6.24 21.5 ± 5.21 24.8 ± 0.70 23.2 ± 2.24
11.02 ± 0.02 - - 1.02 ± 0.02 3.11 ± 0.02 - 1.68 ± 0.01 1.14 ± 0.01
20.93 ± 0.01 - 0.99 ± 0.03 0.75 ± 0.0 2.71 ± 0.03 1.25 ± 0.02 1.62 ± 0. 0 1.11 ± 0.0
Vanillic acid 30.72 ± 0.00 - 1.19 ± 0.01 - 2.36 ± 0.00 1.28 ± 0.04 1.68 ± 0.02 1.45±0.02
40.81 ± 0.04 - - 0.88 ± 0.0 2.36 ± 0.00 1.37 ± 0.01 1.64 ± 0.02 1.09 ± 0.01
A e age 0.87 ± 0.13 - 1.09 ± 0.14 0.88 ± 0.13 2.63 ± 0.35 1.3 ± 0.06 1.65 ± 0.03 1.19 ± 0.16
1ND 0.14 ± 0.01 0.13 ± 0.01 ND 2.81 ± 0.21 1.40 ± 0.01 2.35 ± 0.01 0.56 ± 0.0
2ND - 0.19 ± 0.02 0.60 ± 0.02 2.64 ± 0.01 0.75 ± 0.01 1.53 ± 0.01 0.49 ± 0.0
Ca eic acid* 3- 2.10 ± 0.39 3.58 ± 0.0 0.61 ± 0.01 2.34 ± 0.17 1.68 ± 0.01 2.28 ± 0.01 0.52±0.004
40.05 ± 0.00 - 0.23 ± 0.01 0.34 ± 0.01 2.34 ± 0.17 10.4 ± 0.33 2.13 ± 0.01 4.08 ± 0.01
A e age - - 1.03 ± 1.69 0.51 ± 0.15 2.53 ± 0.23 3.6 ± 4.59 2.07 ± 0.37 1.41 ± 1.77
12.26 ± 0.00 - 2.31 ± 0.13 ND 4.05 ± 0.00 2.40 ± 0.01 2.92 ± 0.02 ND
22.31 ± 0.00 - - ND 4.41 ± 0.00 2.40 ± 0.01 2.91 ± 0.01 ND
Fe ulic acid 3- - - ND 4.04 ± 0.08 2.42 ± 0.01 2.92 ± 0.15 ND
42.27 ± 0.00 - - ND 4.04 ± 0.08 2.42 ± 0.01 2.86 ± 0.01 ND
A e age 2.28 ± 0.02 - . - 4.13 ± 0.18 2.41 ± 0.01 2.90 ± 0.02 ND
12.56 ± 0.01 2.71 ± 0.01 2.58 ± 0.02 2.71 ± 0.08 7.09 ± 0.06 3.65 ± 0.01 5.23 ± 0.05 4.25 ± 0.05
22.48 ± 0.00 2.52 ± 0.01 2.50 ± 0.02 2.62 ± 0.01 9.23 ± 0.04 5.22 ± 0.01 5.28 ± 0.03 5.58 ± 0.01
Sinapic acid 32.47 ± 0.02 2.59 ± 0.01 2.49 ± 0.14 2.74 ± 0.01 3.94 ± 0.02 3.03 ± 0.00 4.92 ± 0.03 4.18±0.10
42.50 ± 0.00 2.56 ± 0.04 2.63 ± 0.01 7.40 ± 1.20 2.76 ± 0.01 5.36 ± 0.01 4.19 ± 0.05
A e age 2.50 ± 0.04 2.61 ± 0.01 2.53 ± 0.04 2.67 ± 0.05 6.91 ± 0.19 3.66 ± 1.10 5.19 ± 0.19 4.55 ± 0.68
13.43 ± 0.01 3.29 ± 0.01 3.56 ± 0.01 3.37 ± 0.00 3.95 ± 0.01 10.4 ± 0.08 6.93 ± 0.01 5.34 ± 0.01
23.44 ± 0.01 3.32 ± 0.01 3.46 ± 0.02 3.45 ± 0.01 5.12 ± 0.00 9.49 ± 0.09 3.83 ± 0.01 5.82 ± 0.01
Ru in 33.64 ± 0.01 3.32 ± 0.01 3.41 ± 0.17 3.41 ± 0.04 3.94 ± 0.02 7.19 ± 0.09 3.56 ± 0.01 6.11±0.02
43.40 ± 0.00 3.31 ± 0.01 3.50 ± 0.01 3.49 ± 0.01 3.95 ± 0.02 8.84 ± 0.04 4.51 ± 1.61 7.25 ± 0.05
A e age 3.47 ± 0.01 3.31 ± 0.01 3.48 ± 0.06 3.43 ± 0.05 4.24 ± 0.58 8.97 ± 1.35 4.51 ± 1.61 6.13 ± 0.81
1- ND 0.93 ± 0.02 0.86 ± 0.01 1.19 ± 0.00 0.84 ± 0.01 1.00 ± 0.01 1.42 ± 0.01
2- ND ND 0.56 ± 0.48 0.86 ± 0.00 0.85 ± 0.00 0.96 ± 0.02 1.38 ± 0.03
Res e a ol 30.96 ± 0.01 - 0.93 ± 0.01 - 0.97 ± 0.14 - 1.05 ± 0.002 1.29±0.01
40.84 ± 0.01 - 0.88 ± 0.03 - 0.97 ± 0.14 - 0.98 ± 0.01 1.32 ± 0.0
A e age 0.90 ± 0.08 - 0.91 ± 0.02 0.71 ± 0.21 0.99 ± 0.13 0.84 ± 0 0.99 ± 0.03 1.35 ± 0.05
1- ND ND 4.90 ± 0.17 ND ND
2- ND ND 4.57 ± 0.09 ND ND
Cinnamic acid 3- - ND 4.66 ± 0.02 ND ND
4- ND ND 4.18 ± 0.57 - ND
A e age - - - 4.57 ± 0.29 - ND ND
15.47 ± 0.04 5.43 ± 0.51 5.14 ± 0.32 5.71 ± 0.02 3.63 ± 0.01 3.58 ± 0.00 3.57 ± 0.03 3.65 ± 0.01
25.20 ± 0.14 5.13 ± 0.28 5.01 ± 0.18 6.01 ± 0.02 3.59 ± 0.00 3.57 ± 0.02 3.62 ± 0.02 3.67 ± 0.01
Que ce in** 35.02 ± 0.16 9.39 ± 0.61 4.92 ± 0.17 5.44 ± 0.02 2.14 ± 0.00 2.04 ± 0.00 3.56 ± 0.001 3.63±0.003
45.05 ± 0.01 4.63 ± 0.01 4.69 ± 0.09 5.36 ± 1.00 2.14 ± 0.01 3.57 ± 0.00 3.59 ± 0.004 3.69 ± 0.0
A e age 5.18 ± 0.21 6.14 ± 2.18 4.94 ± 0.18 5.63 ± 0.29 2.87 ± 0.84 3.19 ± 0.76 3.58 ± 0.02 3.66 ± 0.02
*ca eic acid – de ec ed a 275 nm, **que ce in - de ec ed a 375 nm, p-couma ic acid was no de ec ed in any sample, ND – no de ec ed
Con en o phenolic compounds in wine samples de e mined by HPLC in G uene Vel line (GV) and Zweigel ebe (ZW) o ou ineya ds
(P, S, O B) in Czech Republic (P, S) and Aus ia (O, B) collec ed om 4 di e en sampling loca ions in each ineya d measu ed in
iplica es.
682
B. Soyollkham e al.
wi h in e als app oxima ely 0.6-2.9 mmol L-1 o
whi e wines and alues wi h in e al app oxima ely
3-9 mmol L-1 (TE) o ed wines. These alues we e abou
3-15 imes highe han ou alues, and, o he FRAP
me hod, ou alues we e lowe by 30-90% in ed wines,
and 50-80% in whi e wines, espec i ely [27]. Bee e
al. [32] p esen ed he alues o DPPH in he ange om
0.8-1.06 mmol L-1 TE o whi e wines. Ou alues we e
lowe han one hal compa ed o hese alues. The
di e ences could be caused by he di e en s anda d
used in he expe imen s.
3.3. Indi idual phenolic con en
The ollowing compounds we e iden i ied in he wine
samples (see Fig. 6): he phenolic acids (i.e., gallic acid,
anillic acid, ca eic acid, p-couma ic acid, e ulic acid,
sinapic acid and cinnamic acid), ca echin, es e a ol,
que ce in and u in. Con en s o he indi idual phenolics
in wines om ou di e en wine-p oducing sub-
egions o he Czech Republic and Aus ia a e epo ed
in Table 2.
Figu e 4. To al an ioxidan ac i i ies in whi e wine samples de e mined by he DPPH and FRAP me hods (GAE mmol L-1, SDs see Table 2)
Figu e 5. To al an ioxidan ac i i ies in ed wine samples de e mined by he DPPH and FRAP me hods (GAE mmol L-1, SDs see Table 2)
683
To al polyphenolic compounds con en s (TPC), o al an ioxidan
ac i i ies (TAA) and HPLC de e mina ion o indi idual polyphenolic
compounds in selec ed Mo a ian and Aus ian wines
The esul s ob ained con i med a a ia ion in he
phenolic con en s among wines es ed due o hei
di e en geog aphical o igin. A compa ison o hese
esul s wi h li e a u e alues was di icul because o he
sample p epa a ion and because he ch oma og aphic
condi ions signi ican ly di e ed. Mo eo e , he con en
da a o indi idual phenolics we e limi ed o a ew
compounds and samples in his s udy. Howe e , he
ob ained esul s we e in ag eemen wi h he alues
epo ed in li e a u e. Gallic acid was he mos abundan
phenolic compound (mean 5.69 mg L-1) in whi e wines;
he highes le el (6.19 mg L-1) was ound in OGV-4
sample om he Wein ie el ineya d, while he lowes
amoun (5.04 mg L-1) o gallic acid was ound in SGV-4
sample om he Velkopa lo ická ineya d. Resul s we e
compa ed o he p e iously published da a by Malo aná
e al. [33], Ras ija e al. [16], and Komes e al. [22],
wi hin he concen a ion ange o gallic acid ( om 5.16 o
28.3 mg L-1) de e mined in samples om he Cana y
Islands, (0.7-8.4 mg L-1) ound in samples om C oa ia and
2.63 mg L-1 om Zago je, espec i ely. Gallic acid (mean
13.1 mg L-1) in ed wine was 3- o 5- imes
and 5 imes lowe han he esul s published o Tu kish
and I alian wines, espec i ely [34,35]. Ca echin,
wi h a mean concen a ion o 7.6 mg L-1, was he
second mos abundan phenolics in whi e wines and
wi h 24.5 mg L-1 also in ed wines; his was om 3-
o 10- imes highe han he C oa ian wines esul s
(mean 2.86 mg L-1) and simila o (mean 25.1 mg L-1) he
esul s o Tu kish ed wines, espec i ely [34,35]. The
highes amoun o anillic acid was ound (in a e age
2.64 mg L-1) o SZW ed wine and he lowes was
(a e age alue 0.87 mg L-1) o SGV (whi e wine). High
alues o anillic acid, anging om 4.66 o 5.22 mg L-1,
we e de ec ed in some ed wines om Tu kish egions
[34]. Mo eo e , low alues o his acid, anging om
0.05 o 0.28 mg L-1, we e ound in Spanish wines [36].
Ca eic acid anged om 0.01 o 10.4 mg L-1 in whi e and
ed wines. These esul s we e simila o esul s in Spain
wines (4.09 mg L-1) and I alian ed wines ( anged om
2.5 o 17.9 mg L-1), espec i ely [35,36]. p-Couma ic acid
and cinnamic acids we e de ec ed excep ionally in some
samples in much lowe amoun s, bu i was di icul o
quan i y hei exac concen a ions. The a e age alues
o e ulic acid anged om 2.28 o 2.31 mg L-1 in whi e
wines and 2.41 - 4.13 mg L-1 in ed ones. These esul s
aligned wi h he esul s by Komes e al. [22], ( anged
om 1.88 o 3.2 mg L-1). Mean concen a ions o sinapic
acid we e 2.55 mg L-1 in whi e wines and 5.07 mg L-1 in
ed wines, espec i ely. Amoun s o u in and que ce in
anged om 3.29 o 10.4 mg L-1 and om 2.04 o
9.39 mg L-1 in whi e and ed wines, espec i ely. Ou
esul s we e in ag eemen wi h alues ob ained by
Malo aná e al. [33] and Ras ija e al. [16]. Res e a ol,
a compound wi h mul iple heal h bene i s, was ound
in all wine samples, excep o PGV. Amoun s we e
compa able wi h he concen a ion anges ound in he
li e a u e [16,37].
The o al phenolic con en signi ican ly co ela ed
wi h he an ioxidan ac i i y and con en s o indi idual
phenolic compounds in he wines in es iga ed
(see Fig. 7 as an example) i he spec um o he
phenolics ( hei ela i e abundance) and ma ix o a
sample we e simila . Fo ins ance, he o al con en s o
Figu e 6. HPLC-UV-VIS DAD ch oma og am o mix u e o polyphenols s anda ds a 210 nm, Supelcosil LC-18-DB column, 30°C, g adien elu ion:
sol en A (95% ( / ) ace oni ile acidi ied wi h 0.35 mL TFAA) and sol en B (50% ( / ) aqueous ace oni ile acidi ied wi h 0.25 mL TFAA),
injec ion olume 10 µL, low a e o 1 mL min-1.
684
B. Soyollkham e al.
phenolic compounds and he an ioxidan ac i i y (DPPH
me hod) alues o S a ia ions g own in Velkopa lo ická,
P a ia ions g own in Velké Hos ě ádky - Bošo ice, O
a ia ions g own in Wein ie el and B a ia ions g own
in Wag am showed a good co ela ion. These esul s
ag ee wi h he ela ionship be ween he o al phenolic
con en and he o al an ioxidan po en ial alues o
wines (R2 = 0.9876) by Minussi e al. [30]. Mo eo e ,
in SGV and SZW, he o al an ioxidan ac i i ies o
wines in es iga ed co ela ed well wi h gallic acid
(R2 = 0.95), ca echin (R2 = 0.81), anillic acid (R2 = 0.97),
e ulic acid (R2 = 0.94), sinapic acid (R2 = 0.73), u in
(R2 = 0.56), que ce in (R2 = 0.62) and es e a ol
(R2 = 0.39), espec i ely. In PGV and PZW, he o al
an ioxidan ac i i ies o wines in es iga ed well co ela ed
wi h gallic acid (R2 = 0.98), ca echin (R2 = 0.74),
sinapic acid (R2 = 0.37), u in (R2 = 0.93) and que ce in
(R2 = 0.35), espec i ely. In OGV and OZW, he o al
an ioxidan ac i i ies o wines in es iga ed co ela ed wi h
gallic acid (R2 = 0.97), ca echin (R2 = 0.98), anillic acid
(R2
= 0.99), ca eic acid (R2
= 0.44), u in (R2
= 0.19), que ce in
(R2 = 0.88) and es e a ol (R2 = 0.22), espec i ely.
In BGV and BZW, he o al an ioxidan ac i i ies o
in es iga ed wines co ela ed wi h gallic acid (R2 = 0.95),
ca echin (R2 = 0.91), anillic acid (R2 = 0.37), sinapic acid
(R2 = 0.91), u in (R2 = 0.92), que ce in (R2 = 0.74) and
es e a ol (R2 = 0.82), espec i ely. Ou esul s we e in
ag eemen wi h some in es iga ions p esen ed in he
li e a u e. Acco ding o F ankel e al. [38], he ela i e
an ioxidan ac i i y o 20 selec ed Cali o nian wines
co ela ed wi h o al con en s o phenolics in wines
(R2 = 0.94), and concen a ion o gallic acid (R2 = 0.92),
ca echin (R2 = 0.75), que ce in (R2 = 0.68), ca eic acid
(R2 = 0.63) and u in (R2 = 0.50). Sánchez-Mo eno e al.
[39] obse ed ha he ee adical-sca enging ac i i y
o gallic acid was he highes ; annic acid, ca eic [acid,
que ce in and u in ac i i ies we e in e media e and
e ulic acid and es e a ol we e he lowes .
4. Conclusions
The highes TPC alues we e ound o ed wine
samples (1230 mg L-1) g own in Velkopa lo ická, Czech
Repuiblic. The lowes TPC alue (564 mg L-1) was ound
o ed wine samples g own in Wein ie el (app oxima ely
hal o he alue), Aus ia.The TPC alues o whi e wine
samples we e ela i ely high bu se e al imes lowe
han compa ed o ed wines. The lowes TPC alue was
ound in he whi e wine g own in Wag am in con as o
he ed wine samples om he same egion.
The highes TAA by he FRAP me hod was ound in
ed wines (3.5 - 3.8 mmol L-1, i.e., 7 o 11- imes highe
han ha in whi e wines) while he lowes TAA alue
(1.41 mmol L-1), lowe han hal o he highes alue,
was ound in a ed wine g own in Velké Hos ě ádky
- Bošo ice. The highes TAA by applying he DPPH
me hod was ound in a ed wine (2.0 mmol L-1, i.e., s ill
5- o 8- imes highe han ha ound in whi e wines).
Gallic acid was he mos abundan phenolic
compound (mean 5.7 mg L-1) in whi e wines; he highes
le el (6.2 mg L-1) was ound in a whi e wine sample om
Wein ie el, while he lowes amoun (5.04 mg L-1) o gallic
acid was ound in a sample om he Velkopa lo ická
egion. Ca echin, wi h a mean concen a ion
o 7.6 mg L-1, was he second mos abundan phenolics
in whi e wines, and wi h 24.5 mg L-1, also in ed wines.
The highes concen a ion o anillic acid was ound
(in a e age 2.6 mg L-1) o ed wines and he lowes
was (a e age alue 0.9 mg L-1) o SGV whi e wines.
The ca eic acid concen a ion anged om 0.01 o
10.4 mg L-1 in whi e and ed wines. p-couma ic acid and
cinnamic acids we e de ec ed in some samples in much
lowe amoun s bu i was no possible o exac ly quan i y
hei concen a ions. The a e age concen a ions
o e ulic acid and sinapic acid we e 2.3 mg L-1 and
2.6 mg L-1 in whi e wines and 2.4 - 4.1 mg L-1 and
5.1 mg L-1 in ed wines, espec i ely. Amoun s o u in
and que ce in anged om 3.3 o 10.4 mg L-1 and om
2.0 o 9.4 mg L-1 in whi e and ed wines, espec i ely.
Acco ding o he esul s, i was concluded ha i) he
o al con en o phenolics and o al an ioxidan ac i i y/
o al an ioxidan capaci y and he concen a ion o
phenolic compounds could be a indica o o possible
iden i ica ion o wines’ geog aphical o igin, ii) he o al
con en s o phenolic compounds signi ican ly co ela ed
wi h he an ioxidan ac i i y and he con en s o indi idual
phenolic compounds i he spec um o phenolics ( ela i e
abundance) and he ma ix we e simila , iii) he gallic acid
was he mos abundan compound; annic acid, ca eic
acid, que ce in and u in ac i i ies we e in e media e;
and he e ulic acid and es e a ol showed he lowes
in luence on he ee adical-sca enging ac i i y.
Figu e 7. Rela ionship be ween he o al polyphenolic compounds
con en s (TPC) and an ioxidan ac i i y (TAA) by DPPH
me hod alues o SGV and SZW (as an example). (♦)
whi e wine (●) ed wine
685