Molecules 2010, 15, 4467-4477; doi:10.3390/molecules15064467
molecules
ISSN 1420-3049
www.mdpi.com/jou nal/molecules
A icle
An ioxidan P ope ies o Eu opean C anbe ybush F ui
(Vibu num opulus a . edule)
O aka Rop 1,*, Voj ech Reznicek 2, Magdalena Valsiko a 3, Tunde Ju iko a 4, Ji i Mlcek 1 and
Daniela K ama o a 5
1 Depa men o Food Technology and Mic obiology, Facul y o Technology, Tomas Ba a Uni e si y
in Zlin, Names i T. G. Masa yka 275, 762 72 Zlin, Czech Republic; E-Mail: [email p o ec ed] (J.M.)
2 Depa men o B eeding and P opaga ion o Ho icul u al Plan s, Facul y o Ho icul u e, Mendel
Uni e si y in B no, Val icka 337, 691 44 Lednice, Czech Republic;
E-Mail: [email protected] (V.R.)
3 Depa men o Vege ables-P oduc ion, Ho icul u e and Landscape Enginee ing Facul y, Slo ak
Uni e si y o Ag icul u e in Ni a, Tulipano a 7, 949 76 Ni a, Slo akia;
E-Mail: magd[email p o ec ed] (M.V.)
4 Depa men o Na u al and In o ma ics Sciences, Facul y o Cen al Eu opean S udies, Cons an ine
he Philosophe Uni e si y in Ni a, D azo ska 4, 949 74 Ni a, Slo akia;
E-Mail: [email p o ec ed] (T.J.)
5 Depa men o Food Biochemis y and Analysis, Facul y o Technology, Tomas Ba a Uni e si y in
Zlin, Names i T. G. Masa yka 275, 762 72 Zlin, Czech Republic;
E-Mail: [email p o ec ed]b.cz (D.K.)
* Au ho o whom co espondence should be add essed; E-Mail: [email p o ec ed];
Tel.: + 420-576-031129, Fax: + 420-577-210172.
Recei ed: 5 May 2010; in e ised o m: 22 June 2010 / Accep ed: 22 June 2010 /
Published: 23 June 2010
Abs ac : In he li e a u e he e is li le a ailable in o ma ion conce ning Eu opean
c anbe ybush ui (Vibu num opulus a . edule). This plan can be cul i a ed, e en in
ha sh clima ic condi ions, because o i s low en i onmen al demands, and i is possible o
ha es he ui e en in he snow co e . The aim o his s udy was o de e mine he con en
o polyphenolics, an ioxidan ac i i y, la onoids and i amin C in he ui o h ee
cul i a s ´Lening adskaya o bo naya´, ´Souzga´ and ´Taezny ubiny´ o his species. In he
case o polyphenolics, high con en s [up o 8.29 g o gallic acid/kg o esh mass (FM)]
we e obse ed. The 1,1´-diphenyl-2-pic ylhyd azyl (DPPH) and 2,2´-azinobis-3-e hyl-
OPEN ACCESS
Molecules 2010, 15
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benz hiazino-6-sulphonic acid (ABTS) es s we e applied o de e mine an ioxidan ac i i y,
which was also high in compa ison wi h o he ui species. The co esponding co ela ions
be ween he polyphenolic con en and an ioxidan ac i i y we e in case o he DPPH es
2 = 0.88 and o he ABTS es 2 = 0.98. Fo compa ison, he sca enging ac i i y owa ds
eac i e oxygen species (supe oxide anion, hyd oxyl adical and ni ic oxide) was
de e mined by using a 25% ui ex ac o pa icula cul i a s. An ioxidan e iciency was
also assessed using he a li e slice model. Fu he mo e, he con en s o la onoids and
i amin C we e assayed, gi ing alues o 4.89 g/kg and 1.64 g/kg FM, espec i ely. The
wo k should con ibu e o he popula iza ion o his species as a p omising c op plan in
human nu i ion.
Keywo ds: Eu opean c anbe ybush; phenolics; an ioxidan ac i i y; la onoids;
asco bic acid
1. In oduc ion
The species Vibu num opulus L. belongs o he Cap i oliaceae plan amily [1]. The ui s o
Vibu num opulus a . edule (Eu opean c anbe ybush) and a . ilobum Ma sh. (Ame ican
c anbe ybush) a e edible [2]. While he Ame ican c anbe ybush is na i e o No h Ame ica [3], he
plan s o Eu opean c anbe ybush a e ound in Eu ope, No h Asia and No h A ica, and also in he
cen al zone o Russia [4]. The be ies o Eu opean c anbe ybush a e as ingen , he e o e, hey a e
seldom consumed di ec ly, and he ui juice is he bes known p oduc . I is a adi ional d ink in he
Cen al Ana olia egion (Tu key) [5]. Because o i s s ong as ingen as e [4], he ui lesh can be
used in making mixed ui juices [6]. Eu opean c anbe ybush ui is also used locally o p epa ing
jam, jelly, swee mea and ma malade. Since he be ies ha e a lo o na u al pec ins in ea ly au umn
(consump ion ipeness), added pec ins a e no needed o p oduce he p oduc s men ioned abo e [7].
Wha is also in e es ing is ha du ing he win e mon hs i is also possible o ha es he ui s because
o hei esis ance o cold wea he [8]. Al hough he e is a dec ease in he con en o bioac i e
subs ances, he amoun s o d y ma e and suga s inc ease. The e o e, he ui can become a sou ce o
esh ui ound in na u e co e ed in snow [9].
In Russia he be ies a e used in adi ional and olk medicine [7]. The ui a e a sou ce o a ious
biochemical componen s such as asco bic acid and la onoids [10]. The con en o phenolic
compounds has an in lunce on he high an ioxidan ac i i y o be ies [11]. Sagdic e al. [12] no iced
an imic obial ac i i y o me hanolic ex ac s o ui . Simila ly, an imic obial ac i i y was desc ibed in
he case o Eu opean c anbe ybush ui seed oil [13].
Due o he lack o in o ma ion in he li e a u e, he aim o his wo k was o de e mine o al phenolic
con en (TPC), o al an ioxidan ac i i y (TAA), o al la onoid con en (TFC), sca enging ac i i y o
eac i e oxygen species (ROS) – ni ic oxide, supe oxide anion and hyd oxyl adical, sca enging
ac i i y o lipid pe oxida ion as well as asco bic acid con en (AAC) in Vibu num opulus a . edule
ui . Th ee cul i a s o Eu opean c anbe ybush we e used, namely ´Lening adskaya o bo naya´,
´Souzga´ and ´Taezny ubiny´ which a e all Russian in o igin.
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2. Resul s and Discussion
The ui o Eu opean c anbe ybush a e unique because o hei composi ion and a high con en o
subs ances ha ing an ioxidan ac i i y [6]. In ou measu emen e y high con en s o polyphenolics
we e obse ed. The alues o hei con en s anged om 6.80 o 8.29 g ams o gallic acid/kg FM.
These con en s we e highe han he alues ound in Eu opean c anbe ybush by Akbulu e al. [14].
These au ho s obse ed 3.25 g o gallic acid/kg FM. Fo mos ui species lowe con en s a e ypical
[15]. Fo example, in apples he common con en is be ween 0.6 o 2.1 g ams o gallic acid/kg FM [16]
and in plums i is 2.2 o 5.0 g ams o gallic acid/kg FM [17]. The esul s o he measu emen s
pe o med in he h ee cul i a s showed a iabili y o he o he chemical compounds. In case o
i amin C, i s con en s anged om 1.01 o 1.64 g ams/kg FM. The ch oma og am wi h he ma ked
e en ion ime is shown in Figu e 1. In compa ison wi h abula ed da a o o he ui species his alue
is high, ne e heless, i does no each he alues which a e obse ed in, o example, sea-buck ho n
(Hippophae hamnoides L.), whe e an ex eme alue o 12 g ams o asco bic acid/kg FM can be ound
[15], as poin ed ou byE cisli e al., o example [18]. In e es ingly, i amin C was he only ac o
whe e s a is ical signi icance was demons a ed be ween he yea s (see Table 1).
Figu e 1. Ch oma og am ep esen ing he peak o asco bic acid ( e en ion ime 1.872 min.).
Table 1. To al la onoid and asco bic acid con en s (g/kg FM) o ui s o di e en
cul i a s o Eu opean c anbe ybush (Vibu num opulus a . edule Ma sh.), n = 25.
Cul i a
Yea
TFC
AAC
Lening adskaya
o bo naya
2007 3.85 ± 0.17
a
1.14 ± 0.11
a
2008 3.96 ± 0.16
a
1.35 ± 0.09
b
2009 4.06 ± 0.20
a
1.47 ± 0.08
b
Souzga 2007 3.14 ± 0.21
b
1.01 ± 0.10
a
2008 3.44 ± 0.14
b
1.12 ± 0.06
a
2009 3.25 ± 0.15
b
1.50 ± 0.10
b
Taezny ubiny 2007 4.54 ± 0.25
c
1.64 ± 0.08
b
2008 4.75 ± 0.27
c
1.15 ± 0.11
a
2009 4.89 ± 0.21
c
1.50 ± 0.05
b
Di e en supe sc ip s in each column indica e he signi ican di e ences in he mean a P < 0.05.
Molecules 2010, 15
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Eu opean c anbe ybush ui a e a epo ed o be a good sou ce o la onoids [19], which was also
con i med in ou measu emen s. The con en o la onoids anged om 3.14 o 4.89 g/kg FM (Table
2). In o he ui lowe alues a e common [20].
Table 2. Sca enging e ec o Eu opean c anbe ybush ui (Vibu num opulus a . edule
Ma sh.) ex ac (25%) on ni ic oxide (pe cen age o inhibi ion), supe oxide anion
(pe cen age o inhibi ion), hyd oxyl adical (pe cen age o inhibi ion) and lipid
pe oxida ion (pe cen age o inhibi ion), n = 25.
Cul i a Yea
Ni ic oxide
(%)
Supe oxide
anion (%)
Hyd oxyl adical
(%)
Lipid
pe oxida ion (%)
Lening adskaya
o bo naya
2007
24.16 ± 0.26
a
27.11 ± 0.56
a
22.02 ± 0.85
a
12.71 ± 0.56
a
2008
23.84 ± 0.32
a
27.60 ± 0.39
a
21.86 ± 0.79
a
12.95 ± 0.29
a
2009
24.31 ± 0.29
a
27.55 ± 0.50
a
21.90 ± 0.69
a
13.16 ± 0.28
a
Souzga
2007
22.11 ± 0.15
b
25.16 ± 0.49
b
19.40 ± 0.80
b
11.20 ± 0.36
b
2008
22.10 ± 0.28
b
25.44 ± 0.50
b
19.65 ± 0.75
b
11.62 ± 0.38
b
2009
21.89 ± 0.30
c
25.13 ± 0.45
b
19.61 ± 0.81
b
11.51 ± 0.40
b
Taezny ubiny
2007
25.19 ± 0.31
d
28.50 ± 0.38
c
23.94 ± 0.74
c
13.78 ± 0.51
ac
2008
25.44 ± 0.24
d
27.95 ± 0.30
ac
23.78 ± 0.68
c
13.90 ± 0.37
c
2009
25.37 ± 0.26
d
28.15 ± 0.41
c
23.98 ± 0.80
c
13.45 ± 0.42
ac
Di e en supe sc ip s in each column indica e he signi ican di e ences in he mean a P < 0.05.
Conside ing he ac ha all cul i a s we e g own unde iden ical condi ions and in he same
locali y, i is possible o conclude ha one can clea ly see cul i a a iabili y, which is qui e ypical o
ui [21]. This a iabili y became pa icula ly e iden in case o o al an ioxidan ac i i y (see Table
3), whe e wo me hods we e used o i s measu emen [ he 1,1´-diphenyl-2-pic ylhyd azyl (DPPH) and
2,2´-azinobis-3-e hylbenz hiazino-6-sulphonic acid (ABTS) es s].
Table 3. To al phenolic con en (g ams o gallic acid/kg FM) and an ioxidan ac i i y
(g ams o AAE/kg FM) o ui s o di e en cul i a s o Eu opean c anbe ybush
(Vibu num opulus a . edule Ma sh.), n = 25.
Cul i a Yea TPC TAA (DPPH
es )
TAA (ABTS
es )
Lening adskaya
o bo naya
2007 7.20 ± 0.22a 9.21 ± 0.16a 9.90 ± 0.14a
2008 7.27 ± 0.15a 9.28 ± 0.17a 9.95 ± 0.18a
2009 7.29 ± 0.20a 9.34 ± 0.10a 9.98 ± 0.17a
Souzga
2007 6.80 ± 0.15b 8.58 ± 0.15b 9.10 ± 0.20b
2008 6.86 ± 0.12ab 8.66 ± 0.15b 9.17 ± 0.18b
2009 6.80 ± 0.15b 8.55 ± 0.30b 9.14 ± 0.23b
Taezny ubiny
2007 8.14 ± 0.14c 9.59 ± 0.13c 10.94 ± 0.31c
2008 8.31 ± 0.21c 9.79 ± 0.19c 11.12 ± 0.26c
2009 8.29 ± 0.22c 9.75 ± 0.14c 11.02 ± 0.25c
Di e en supe sc ip s in each column indica e he signi ican di e ences in he mean a P < 0.05.
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Highe an ioxidan ac i i y was de e mined by he ABTS es . In he ´Souzga´ cul i a i was 9.14
g ams o AAE/kg FM in a h ee-yea a e age, in he ´Lening adskaya o bo naya´ cul i a i was 9.94
g ams o AAE/kg FM. An ioxidan ac i i y was he highes in he ´Taezny ubiny´ cul i a wi h a
h ee-yea a e age alue o 11.01 g ams o AAE/kg FM. In o he ui species, including small be ies
and s one ui , he alues o an ioxidan ac i i y a e usually much lowe [15]. Fo example, in che ies
alues o up o 0.9 g ams o AAE/kg FM a e seen [22] and in plums he alues can each up o 6 g ams
o AAE/kg FM [17]. As a as s a is ical e alua ion o he esul s is conce ned, he highes alues o
he co ela ion coe icien be ween an ioxidan ac i i y and he o al amoun o phenolic subs ances
we e ob ained (in case o he DPPH es 2 = 0.88, y = 1.2158x - 3.7383; in case o he ABTS es
2 = 0.98, y = 0.7646x - 0.2328). Many au ho s ha e no iced a high co ela ion be ween TPC and
an ioxidan ac i i y in ui [16,23].
The ui ex ac s (25%) showed mode a e inhibi o y abili y on ni ic oxide (21.89–25.44%),
supe oxide anion (25.13–28.50%), hyd oxyl adical (19.40–23.94%) and lipid pe oxida ion
(11.20–13.90%). Eu opean c anbe ybush ui was mo e e ec i e han o he ui species, e.g.,
mulbe y [24] o apples [25]. ROS a e gene a ed in li ing o ganisms du ing me abolism. Excessi e
amoun s o ROS may be ha m ul because hey can ini ia e biomolecula oxida ions which lead o cell
inju y [26]. ROS a e implica ed in he pa hophysiology o diseases, such as cance , heuma oid
a h i is, ci hosis and a e ioscle osis as well as in he degene a i e p ocess associa ed wi h ageing
[27]. Lipid pe oxida ion is o en caused by ROS as an oxida i e al e a ion o polyunsa u a ed a y
acids. In a biological sys em, lipid pe oxida ion gene a es a numbe o deg ada ion p oduc s and i is
ound o be an impo an cause o cell memb ane des uc ion [28].
3. Expe imen al
3.1. Desc ip ion o g owing locali y
F ui we e ha es ed in an expe imen al gene- und o cha d o Mendel Uni e si y in B no o e he
pe iod om 2007–2009. This o cha d is si ua ed in he a ea o he illage called Zabcice,
app oxima ely 20 km sou h o B no, in he Czech Republic. The al i ude is 184 m. The a e age annual
empe a u e and a i y-yea a e age sum o p ecipi a ion a e 9 ºC (du ing he g owing season 15.6 ºC)
and 553 mm (du ing he g owing season 356 mm), espec i ely. The soils a e classi ied as gleyed
allu ial soils de eloped on he Holocene calci e ous sedimen s wi h a ma ked accumula ion o o ganic
compounds. As a as he ex u e is conce ned, he opsoil is loamy and he subsoil clayey-loamy [29].
3.2. Sample p ocessing
F ui we e ha es ed in Sep embe om i e plan s o each cul i a unde s udy wi hin he pe iod o
consump ion ipeness [7] ( hus each yea had 5 eplica es). Twen y andomly chosen ui s om each
ee we e used o analyses (i.e., al oge he 100 pe each cul i a ). The cul i a s ´Lening adskaya
o bo naya´, ´Souzga´ and ´Taezny ubiny´ we e used. Thei co esponding ui colou s a e in case o
´Lening adskaya o bo naya´ and ´Taezny ubiny´ b igh ed, and as a as he ´Souzga´ cul i a is
conce ned, i s colou is a he da k ed.

Molecules 2010, 15
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The ui o indi idual ees we e p ocessed a e he ha es (no la e han wi hin wo days).
Ha es ed ui s we e pu éed in a mixe and an a e aged sample was ob ained by di iding in o
qua e s. Each pa ame e was measu ed in i e eplica es om he ui aken om each ee o
pa icula cul i a s (n = 25).
3.3. To al phenolic con en (TPC) and an ioxidan ac i i y (TAA) assay
The ex ac ion was pe o med acco ding o he me hod desc ibed by Kim e al. [21], using 10 g o a
esh sample homogenized o 10 seconds in an ex ac ion mix u e o hyd ochlo ic acid-me hanol-
wa e in a a io o 2:80:18. Folin-Ciocal eau eagen was used o measu emen o TPC. A sample (0.5
mL) was aken and dilu ed wi h wa e in a 50 mL olume ic lask. The ea e , Folin-Ciocal eau
eagen (2.5 mL) and 20-pe cen solu ion o sodium ca bona e (7.5 mL) we e added. The esul ing
abso bance was measu ed on a LIBRA S6 spec opho ome e a a wa eleng h o 765 nm agains a
blank sample, which was used as e e ence. The esul s we e exp essed as g ams o gallic acid
(GAE)/kg o esh mass (FM).
An ioxidan ac i i y was measu ed using he ABTS (2,2´-azinobis-3-e hylbenz hiazino-6-sulphonic
acid) me hod desc ibed by Sulc e al. [30]. ABTS (54.9 mg) we e dissol ed in phospha e bu e
(20 mL, pH 7.0; c = 5 mmol/L) and ac i a ed o he ABTS+ ca ion adical o m by means o an
addi ion o o MnO2 (1 g). The esul ing solu ion was in e mi en ly s i ed o an ac i a ion pe iod o
30 min. The ea e , he solu ion was cen i uged o 5 min. and a 7,000 .p.m. and il e ed h ough a
sy inge il e (0.25 µm) and a po ion o he il a e (2 mL) was dilu ed wi h phospha e bu e o an
abso bance o 0.500 ± 0.01, which was measu ed a he wa eleng h o 734 nm. A e he abso bance
was measu ed, he ui ex ac (0.5 mL) was added and he new abso bance alue was measu ed a e
20 minu es.
The DPPH (2,2-diphenyl-1-pic ylhyd azyl) assay was done acco ding o he me hod o B and-
Williams e al. [31] wi h some modi ica ions [32]. The s ock solu ion was p epa ed by dissol ing
DPPH (24 mg) in me hanol (100 mL) and hen s o ed a −20 °C un il needed. The wo king solu ion
was ob ained by mixing he s ock solu ion (10 mL) wi h me hanol (45 mL) o ob ain an abso bance o
1.1 ± 0.02 uni s a 515 nm using he LIBRA S6 spec opho ome e . The ui ex ac s (150 µL) we e
allowed o eac wi h he DPPH solu ion (2,850 µL) o 1 hou in he da k. The ea e , he abso bance
was aken a 515 nm. The an ioxidan ac i i y was calcula ed as a dec ease in he abso bance alue
using he o mula:
An ioxidan ac i i y (%) = (A0 – A1/A0) × 100%
whe e A0 is he abso bance o he con ol (wi hou he sample) and A1 is he abso bance o he mix u e
con aining he sample. The abso bance esul s we e con e ed using a calib a ion cu e o he s anda d
and exp essed in asco bic acid equi alen s (AAE) [33].
3.4. Reac i e oxygen species sca enging ac i i y assay
Fo he measu emen o eac i e oxygen species ac i i y a 25% ui ex ac was p epa ed in
phospha e bu e (c = 50 mmol/L, pH 7.0). The hyd oxyl adical sca enging ac i i y was assayed
acco ding o he me hod by Ghiselli e al. [34]. The ex ac (1 mL) was mixed wi h eac ion bu e
Molecules 2010, 15
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(0.8 mL, KH2PO4·KOH, c = 0.2 mol/L, pH 7.4; deoxy ibose, c = 1.75 µmol/L; i on ammonium
sulpha e, c = 0.1 µmol/L; and EDTA, c = 0.1 µmol/L). H2O2 (0.1 mL, c = 0.01 mol/L) was hen added
o he eac ion solu ion. The solu ion was incuba ed o 10 min. a 37 ºC p io o he addi ion o 1%
hioba bi u ic acid (0.5 mL) and 2.8% ichlo ace ic acid (1 mL). The mix u e was boiled o 10 min
and cooled apidly. The abso bance o he mix u e was measu ed a 532 nm (LIBRA S6 appa a us).
The ni ic oxide sca enging ac i i y assay was done acco ding o he me hod desc ibed by G een e
al. [35]. The ex ac (1 mL) was mixed wi h he eac ion solu ion (1 mL) con aining sodium
ni op usside (c = 10 mmol/L) in phospha e bu e (c = 50 mmol/L, pH 7.0). Incuba ion a 37 ºC o
1 h ollowed and an aliquo (0.5 mL) was hen mixed wi h G iess’ eagen (0.5 mL). The abso bance a
540 nm was measu ed.
The supe oxide anion sca enging ac i i y assay was done by he me hod desc ibed by Beissenhi z
e al. [36] and i is based on he educ ion o cy och ome c. The ex ac (1 mL) was mixed wi h a
solu ion (1 mL) o con aining 0.07 uni s pe mL o xan hine oxidase, xan hine (c = 100 µmol/L) and
cy och ome c (c = 50 µmol/L). A e he incuba ion a 20 ºC o 3 min, he abso bance a 550 nm was
de e mined.
All es s we e pe o med in iplica e. The sca enging ac i i ies o hyd oxyl adical, ni ic oxide and
supe oxide anion we e calcula ed as ollows:
Sca enging ac i i y (%) = (A0 – A1/A0) × 100%
whe e A0 is he abso bance o he con ol (wi hou he sample) and A1 is he abso bance o he mix u e
con aining he sample.
3.5. Lipid pe oxida ion inhibi ion ac i i y
The inhibi ion o lipid pe oxida ion was assayed by he me hod by S i as a a e al. [37]. Ra li e
(5 µg) was homogenized in T is-HCl bu e (20 mL, c = 40 mmol/L, pH 7.0). The li e homogena e
(0.1 mL) was incuba ed wi h he sample (0.2 mL o a 25% ex ac ), KCl (0.1 mL, c = 30 mmol/L),
FeSO4 (0.1 mL, c = 0.16 mmol/l) and asco bic acid (0.1 mL, c = 0.06 mmol/L) a 37 ºC o 1 h.
The ea e , 1% hioba bi u ic acid (TBA, 1 mL) and 15% ichlo ace ic acid (1 mL) we e added. The
inal solu ion was hea ed a 100 ºC in a boiling wa e ba h o 15 min, cooled wi h ice o 10 min, and
hen cen i uged a 5,000 .p.m. o 10 min. The abso bance o he supe na an was measu ed a 532
nm, using he LIBRA S6 spec opho ome e . The blank was made up by subs i u ing T is-HCl bu e
(c = 50 mmol/L, pH 7.0) o he sample. The inhibi ion pe cen age o he o ma ion o TBA- eac i e
subs ances was calcula ed as:
Inhibi ion ac i i y (%) = (A0 – A1/A0) × 100%
whe e A0 is he abso bance o he con ol (wi hou he sample) and A1 is he abso bance o he mix u e
con aining he sample.
3.6. To al la onoid con en assay
The o al la onoid con en was de e mined acco ding o Single on e al. [38]. In a 10 mL
Eppendo ube, he ui ex ac (0.3 mL), 30% e hanol (3.4 mL), NaNO2 (0.15 mL, c = 0.5 mol/L)
Molecules 2010, 15
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and AlCl3·6H2O (0.15 mL, c = 0.3 mol/L) we e added and mixed. A e 5 min, NaOH
(1 mL, c = 1 mol/L) was added, and he mix u e was measu ed a he wa eleng h o 506 nm. The o al
la onoid concen a ion was calcula ed om a calib a ion cu e using u in as he s anda d. The esul s
we e exp essed in g/kg FM.
3.7. Asco bic acid con en assay
The de e mina ion o asco bic acid con en was ca ied ou by a modi ica ion o he me hod by Miki
[39]. The sample (5 g) was ex ac ed in an ex ac ion mix u e (me hanol-H2O-H3PO4 in he a io
99:0.5:0.5). The HPLC sys em used o asco bic acid analysis consis ed o a Supelcosil LC8
ch oma og aphic column (150.0 × 4.6 mm; 5 µm), a Coulochem III elec ochemical de ec o wi h a
gua d cell (ESA Inc., Model 5010A, wo king elec ode po en ial 600 mV and 650 mV), a sol en
deli e y pump (ESA Inc., Model 582). Ch oma og aphic condi ions we e: empe a u e 30 °C, mobile
phase a mix u e o me hanol-H2O-H3PO4 (99:0.5:0.5), isoc a ic elu ion, low a e 1.1 mL/min. The
e en ion ime o asco bic acid was 1.9–2.0 min. The con en o asco bic acid was calcula ed as g·kg-1
o FM.
3.8. S a is ical analysis
The da a ob ained we e analyzed s a is ically by he analysis o a iance (ANOVA) and Tukey´s
mul iple ange es o compa ison o means [40]. Co ela ion unc ions we e calcula ed using he
s a is ical package Unis a , . 5.1 and O ice Excel® Mic oso .
4. Conclusions
In ou measu emen , e y high con en s o polyphenolic subs ances and asco bic acid we e
obse ed in he ui o Eu opean c anbe ybush (Vibu num opulus a . edule Ma sh.). Fu he mo e,
he e was a high co ela ion be ween hese subs ances and an ioxidan ac i i y. F om his poin o
iew, he ui o Eu opean c anbe ybush ha e unique p ope ies in compa ison wi h o he ui
species. They could become a aluable sou ce o nu i ionally impo an subs ances o human
nu i ion. Mo eo e , hey can ha e a signi ican in luence on s eng hening human immuni y and he
p e en ion o many diseases. The wo k deals wi h cul i a a iabili y, which is desc ibed spo adically
in li e a u e and new knowledge is gained on ROS and lipid pe oxida ion sca enging ac i i y o his
ui . In his way he knowledge can suppo immedia e u iliza ion in bo h heo e ical and p ac ical
a eas – in b eeding wo k, selec ion, and e e yday g owing p ac ice.
Acknowledgemen
This wo k was kindly suppo ed by unding om he Minis y o Educa ion, You h and Spo s o
he Czech Republic (g an no. MSM 7088352101).
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