Antioxidant properties of European cranberrybush fruit (Viburnum opulus var. edule)

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 4468 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. Molecules 2010, 15 4469 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 4470 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. Molecules 2010, 15 4471 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 4472 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 4473 (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 4474 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. 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