Phenolic content, antioxidant capacity, radical oxygen species scavenging and lipid peroxidation inhibiting activities of extracts of five black chokeberry (Aronia melanocarpa (Michx.) Elliot) cultivars

Jou nal o Medicinal Plan s Resea ch Vol. 4(22), pp. 2431-2437, 18 No embe , 2010 A ailable online a h p://www.academicjou nals.o g/JMPR ISSN 1996-0875 ©2010 Academic Jou nals Full Leng h Resea ch Pape Phenolic con en , an ioxidan capaci y, adical oxygen species sca enging and lipid pe oxida ion inhibi ing ac i i ies o ex ac s o i e black chokebe y (A onia melanoca pa (Michx.) Ellio ) cul i a s O aka Rop1*, Ji i Mlcek1, Tunde Ju iko a2, Magdalena Valsiko a3, Ji i Socho 4, Voj ech Reznicek4 and Daniela K ama o a5 1Depa 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. 2Depa 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 ak Republic. 3Ho icul u e and Landscape Enginee ing Facul y, Slo ak Uni e si y o Ag icul u e, Tulipano a 7, 949 76 Ni a, Slo ak Republic. 4Depa 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 o Ag icul u e and Fo es y in B no, Val icka 337, 691 44 Lednice, Czech Republic. 5Depa 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. Accep ed 29 Sep embe , 2010 Black chokebe ies (A onia melanoca pa (Michx.) Ellio ) a e species, he ui o which a e conside ed o be one o he mos aluable sou ces o phenolic subs ances and he esea ch in he ield o he ui u iliza ion as a ood supplemen is mo e han opical. Ne e heless, as a as his species is conce ned, in pa icula cul i a s he e a e di e ences which a e caused by bo h hei o igin and gene ic p edisposi ions. The e o e, he aim o ou measu emen was o compa e i e widely g own cul i a s o his plan , namely ´A on´ which is Danish in o igin, ´Fe ödi´ which is Hunga ian in o igin, ´Hugin´ which is o Swedish o igin, ´Ne o´ which is o Czech o igin and ´Viking´ which is Finnish in o igin. The highes con en s o o al phenolics (TPC) we e obse ed in he cul i a ´Viking´ wi h he alue o 12.85 g ams o gallic acid/kg FM and in he cul i a ´Ne o´ he alue eached 11.12 g ams o gallic acid/kg FM. Fu he mo e, in bo h men ioned cul i a s he alues o o al an ioxidan capaci y (TAC) we e he highes . The co ela ion coe icien be ween TPC and TAC was 2 = 0.943. The con en o chlo ogenic and neochlo ogenic acid showed an in luence on TAC ( 2 = 0.788 and 2 = 0.940). Fo compa ison, sca enging ac i i y o eac i e oxygen species (supe oxide anion, hyd oxyl adical and ni ic oxide) was de e mined by using 25% me hanolic ex ac s o ui o pa icula cul i a s. In addi ion, an ioxidan po en ial was assessed using he a li e slice model. Simila ly, he highes alues o sca enging ac i i y we e ound in he cul i a s ´Viking´ and ´Ne o´ which seem o be mos p omising o u he u iliza ion in alimen a y and pha maceu ical p ac ices. Key wo ds: Chokebe y, phenolics, an ioxidan capaci y, ROS, lipid pe oxida ion. INTRODUCTION Chokebe y (A onia melanoca pa (Michx.) Ellio ) is a membe o he Rosaceae amily and ui a e ac ually a pome (Wal he and Schnell, 2009). Nowadays, he ui a e highligh ed wi h espec o hei po en ial as a ood colo an (B idle and Timbe lake, 1997) and as a sui able ood supplemen in ela ion o s eng hening human immuni y sys em (Jakobek e al., 2007). Pe haps he mos impo an cons i uen s ound in chokebe ies a e he phenolic compounds which a e esponsible o many medicinal p ope ies (Kulling and Rawel, 2008). In he 2432 J. Med. Plan . Res. ui , chlo ogenic and neochlo ogenic acids a e dominan among he a oma ic acids (Slimes ad e al., 2005). High con en s o cyanidin-3-a abinoside, cyanidin-3- galac oside and (-)epica echin a e also ypical o chokebe ies (Skupien and Oszmianski, 2007). ROS a e gene a ed in many edox p ocesses in human body and o en induce oxida i e damage o biomolecules. When he gene a ion o ROS induced by a ious s imuli in he o ganism exceeds he an ioxidan capaci y o he o ganism, i will lead o a a ie y o pa hophysiological p ocesses (Yang e al., 2008). ROS can cause oxida ion o biomolecules such as DNA, p o ein and enzymes (Wu and Ng, 2008). ROS can cause oxida ion o biomolecules such as DNA, p o ein and enzymes. ROS also a ack lipids o ini ia e ee adical chain eac ions and cause lipid pe oxida ion (Wu and Ng, 2008). An ioxidan capaci y o phenolics agains lipid pe oxida ion is e y e ec i e and was desc ibed by many au ho s e.g (Takao e al., 1994; Bane jee e al., 2005; G acia-Alonso e al., 2004). In compa ison wi h o he ui species, ela i ely high alues o an ioxidan capaci y we e epo ed in chokebe y ui (Kulling and Rawel, 2008). An ioxidan p ope ies a e asc ibed o a high con en o polyphenols (Wal he and Schnell, 2009). The aim o ou wo k was o measu e TPC, TAC and he con en o cyanidin-3- a abinoside, cyanidin-3-galac oside, (-)epica echin, chlo ogenic and neochlo ogenic acids as impo an phenolic cons i uen s in black chokebe y ui . To suppo he esul s, e iciency o pa icula chokebe y cul i a ex ac s on adical oxygen species (ROS) sca enging ac i i y (hyd oxyl adical, ni ic oxide and supe oxide anion) was measu ed. An ioxidan po en ial was also assessed using he a li e slice model and lipid pe oxida ion sca enging ac i i y was measu ed. The wo k is o p o ide wi h use ul in o ma ion when compa ing he mos widesp ead cul i a s o his ui species (pa icula ly as a as TPC, TAC and ROS a e conce ned). MATERIALS AND METHODS Desc ip ion o locali y F ui we e ha es ed in expe imen al o cha ds o Tomas Ba a Uni e si y in Zlin wi hin he pe iod o 2008 – 2010. These o cha ds a e si ua ed in he sou h-wes e n pa o he Whi e Ca pa hians *Co esponding au ho . E-mail: op@ .u b.cz. Tel: +420 576 031 129. Fax: + 420 577 210 172. Abb e ia ions: AAE, asco bic acid equi alen s; DPPH, 2,2- Diphenyl-1-pic ylhyd azyl; FM, esh mass; GAE, gallic acid; ROS, adical oxygen species; TAC, o al an ioxidan capaci y; TPC, o al phenolic con en . nea Zlin, he Czech Republic. The a e age al i ude is 340 m abo e sea le el, and he mean annual empe a u e and p ecipi a ion a e 7.9°C and 760 mm, espec i ely. The soil ype was classi ied as he Meso ophic Cambisol (Anonymous, 2007). Collec ion and p ocessing o samples o chemical analyses F ui we e ha es ed in ull ipeness (H ico sky, 2002) om h ee plan s o each cul i a unde s udy in he cou se o Augus . 40 andomly chosen ui om each plan we e mixed oge he and used o analyses ( ha is, al oge he 120 pe each cul i a ). F ui o indi idual cul i a s we e p ocessed immedia ely a e he ha es (no la e han wi hin wo days). Ha es ed ui we e pu éed in a mixe and he a e age sample was ob ained by di iding in o qua e s. Each pa ame e was measu ed in i e eplica ions. The esul s we e exp essed as a e age o a h ee-yea expe imen . The ollowing cul i a s o black chokebe ies we e analyzed – ´A on´ which is Danish in o igin, a Hunga ian cul i a ´Fe ödi´, ´Hugin´ which is o Swedish o igin, ´Ne o´ which is a Czech cul i a and ´Viking´ which is Finnish in o igin (Kulling and Rawel, 2008). Sample p epa a ion Ex ac ion was pe o med acco ding o he me hod desc ibed by Kim e al. (2003) and modi ied acco ding o Ba os e al. (2007), using he ollowing p ocedu e: 10 g o a esh sample we e homogenized o 10 seconds in 100 mL o me hanol. The esul ing pas e was placed in o E lenmeye lasks (120 mL) and le o s and in a wa e ba h wi h he empe a u e o +25°C o a pe iod o 24 hou s. The esidue was hen ex ac ed wi h wo addi ional po ions o me hanol. The combined me hanolic ex ac s we e e apo a ed a 40˚C o d yness and edissol ed in me hanol a a concen a ion o 100 mg/mL, and s o ed a 4˚C o u he use. To al phenolic con en assay To measu e o al con en s o phenolic subs ances, 0.5 mL o he sample was aken and dilu ed wi h wa e in a 50 mL olume ic lask. The ea e , 2.5 mL o Folin-Ciocal eau eagen and 7.5 mL o a 20% solu ion o sodium ca bona e we e added. The esul ing abso bance was measu ed in he spec opho ome e LIBRA S6 a he wa eleng h o 765 nm agains a blind sample, which was used as e e ence. The esul s we e exp essed as g o gallic acid/kg o esh mass (FM) (Kim e al., 2003). An ioxidan capaci y by he DPPH es assay The DPPH es (2,2-Diphenyl-1-pic ylhyd azyl) was done acco ding o he me hod o B and-Williams e al. (1995) wi h some modi ica ions (Thaipong e al., 2006). The s ock solu ion was p epa ed by dissol ing 24 mg o DPPH wi h 100 mL o me hanol and hen s o ed a -20°C un il needed. The wo king solu ion was ob ained by mixing 10 mL o he s ock solu ion wi h 45 mL o me hanol o ob ain he abso bance o 1.1 ± 0.02 uni s a 515 nm using he spec opho ome e LIBRA S6. F ui ex ac s (150 µL) we e allowed o eac wi h 2.850 µL o he DPPH solu ion o 1 h in he da k. Then, he abso bance was aken a 515 nm. The an ioxidan capaci y was calcula ed as a dec ease in abso bance alue using he o mula: (%) = (A0 – A1/A0) x 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 esul s o abso bance 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)/kg FM (Rupasinghe e al., 2006). Phenolic con en assay Fo he measu emen o cyanidin-3-a abinoside, cyanidin-3- galac oside, (-)epica echin, chlo ogenic and neochlo ogenic acids he s o ed me hanolic ex ac was dilu ed a hund ed imes wi h ACS wa e , which is deionized wa e supplied by Sigma-Ald ich and il a ed h ough a memb ane il e Nylon (0.45 µm Nylon il e disk). The ins umen used o phenolic analysis consis ed o a sol en deli e y pump (ESA Inc., Model 582), gua d cell (ESA Inc., Model 5010A, wo king elec ode po en ial K1 = 600 mV, K2 = 650 mV), ch oma og aphic column - Model Supelcosil LC8 (150.0 × 4.6 mm), 5 µm pa icle size and an elec ochemical de ec o (Coulochem III). Ch oma og aphic condi ions we e cons an : 30°C, as a mobile phase me hanol was used:H2O:H3PO4 = 99:0.5:0.5, ( il a ed h ough a il e Nylon, 0.2 µm), he ype o elu ion was isoc a ic, he low a e o he mobile phase was 1.1 mL/min (Miki, 1981). The con en o phenols was calcula ed as mg/kg o esh mass. 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 25% ex ac s we e p epa ed in phospha e bu e (20 mmol/L, pH 7.4). The hyd oxyl adical sca enging ac i i y was assayed acco ding o he me hod o Ghiselli e al. (1998). 1 mL o he ex ac was mixed wi h 0.8 mL o a eac ion bu e (phospha e bu e , 20 mmol/L, pH 7.4, deoxy ibose, 1.75 µmol/L, i on ammonium sulpha e, 0.1 µmol/L and EDTA, 0.1 µmol/L). 0.1 ml o H2O2 (0.01 mol/L) was hen added o he eac ion solu ion. The solu ion was incuba ed o 10 min a 37°C p io o he addi ion o 0.5 mL o 1% hioba bi u ic acid and 1 mL o 2.8% ichlo ace ic acid. 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 (appa a us LIBRA S6). The assay o ni ic oxide sca enging ac i i y was done acco ding o he me hod desc ibed by G een e al. (1982). 1 mL o he ex ac was mixed wi h 1 mL o he eac ion solu ion con aining sodium ni op usside (10 mmol/L) in phospha e bu e (20 mmol/L, pH 7.4). The incuba ion a 37°C o 1 h ollowed and 0.5 mL o aliquo was hen mixed wi h 0.5 mL o G iess eagen . The abso bance was measu ed a 540 nm. The supe oxide anion sca enging ac i i y was done acco ding o he me hod desc ibed by Beissenhi z e al. (2004) and i is based on he educ ion o cy och ome c. 1 mL o he ex ac was mixed wi h 1 mL o he solu ion con aining xan hine oxidase (0.07 U/mL), xan hine (100 µmol/L) and cy och ome c (50 µmol/L). A e incuba ion a 20°C 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 hyd oxyl adical sca enging ac i i y, ni ic oxide sca enging ac i i y and supe oxide anion sca enging ac i i y we e calcula ed as ollows: Sca enging ac i i y (%) = (A0 – A1/A0) x 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. Lipid pe oxida ion inhibi ion ac i i y The inhibi ion o lipid pe oxida ion was assayed by he me hod o Anup e al. (2006). 5 µg o a li e we e homogenized in 20 mL o T is-HCl bu e (50 mmol/L, pH 7.6). 0.1 mL o he li e homogena e was incuba ed wi h he sample (0.2 mL o a 25% ex ac ), 0.1 mL o KCl (30 mmol/L), 0.1 ml o FeSO4 (0.16 mmol/L) and 0.1 mL o asco bic acid (0.06 mmol/L) a 37°C o 1 h. The ea e , 1 mL o 1% hioba bi u ic acid (TBA) and 1 mL o 15% ichlo ace ic acid Rop e al. 2433 we e added. The inal solu ion was hea ed a 100°C 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 pm. 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 pe o med by subs i u ing T is- HCl bu e (50 mmol/L, pH 7.6) 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) x 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. 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 (Snedeco and Coch an, 1967). 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 . RESULTS F om ou esul s one can see high a iabili y among he ui o pa icula cul i a s. In case o TPC he highes a e age alues we e ob ained in he ´Viking´ cul i a (12.85 g GAE/kg FM), ollowed by he ´Ne o´ (11.12 g GAE/kg FM) and he ´Hugin´ (9.68 g GAE/kg FM) cul i a s. On he con a y, in he ´Fe ödi´ and ´A on´ cul i a s he alues we e only 7.78 g GAE/kg FM and 7.89 g GAE/kg FM, espec i ely. Simila ly, he alues o TAC co esponded o he measu ed con en s. Fo example, in he ´Viking´ cul i a , TAC was 15.96 g AAE/kg FM and in he ´Ne o´ cul i a he alue was 15.96 g AAE/kg FM. On he o he hand, he lowes amoun o TAC was ound in he ui o he ´Fe ödi´ cul i a wi h he alue being 8.89 g AAE/kg FM (Table 1). The co ela ion coe icien be ween TPC and TAC was 2 = 0.948, y = 1.5194x - 2.9191. In case o pa icula polyphenols s a is ical signi ican di e ences we e obse ed among he cul i a s. The alues o hei con en s anged om 941.82 mg/kg FM ( he ´A on´ cul i a ) o 1553.29 mg/kg FM ( he ´Viking´ cul i a ) in cyanidin-3-a abinose; in cyanidin-3- galac oside om 1010.80 mg/kg FM ( he ´A on´ cul i a ) o 1203.56 mg/kg FM ( he Viking´ cul i a ); in case o (-) epica echin he alues we e om 467.35 mg/kg FM ( he ´A on´ cul i a ) o 862.50 mg/kg FM ( he ´Ne o´ cul i a ). The lowes alues o chlo ogenic and neochlo ogenic acids we e measu ed in he ´Fe ödi´ cul i a (1131.15 mg/kg FM and 845.04 mg/kg FM´). The ch oma og am ep esen ing he pa icula peaks is shown in Figu e 1. The co ela ion coe icien be ween an ioxidan capaci y and chlo ogenic acid was 2 = 0.788, y = 0.0075x – 0.1052. On he con a y, in case o neochlo ogenic acid he co ela ion coe icien was 2 = 0.940, y = 0.0258x –1.3268. Fo compa ison, he co ela ion 2434 J. Med. Plan . Res. Table 1. TPC and TAC con en s – in case o TPC he esul s a e exp essed as g ams o gallic acid/kg FM, in case o TAC he esul s a e exp essed as g ams o AAE/kg FM. Cul i a TPC TAC A on 7.89 ± 0.35a 9.02 ± 0.56a Fe ödi 7.78 ± 0.39a 8.89 ± 0.71a Hugin 9.68 ± 0.44b 11.15 ± 0.88b Ne o 11.12 ± 0.57c 15.32 ± 1.12c Viking 12.85 ± 0.56c 15.96 ± 0.95c 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. coe icien s be ween TAC and cyanidin-3-a abinoside, cyanidin-3-galac oside and (-)epica echin we e 2 = 0.958, y = 0.0116x – 1.6738; 2 = 0.903, y = 0.0397x – 3.1910; and 2 = 0.948, y = 0.0190x – 0.7533, espec i ely. In e es ing esul s we e gi en in case o sca enging e ec on ROS o 25% black chokebe y ui me hanolic ex ac s. Compa ing he pa icula esul s shown in Table 3, high a iabili y be ween he cul i a s is e iden . Gene ally, he lowes alues o sca enging ac i i y we e eached ( he esul s a e lis ed in o de o hyd oxyl adical, ni ic oxide, supe oxide anion and lipid pe oxida ion) in he ´Fe ödi´ cul i a (22.08, 27.59, 21.24, 12.05%) and ha o he ´A on´ (25.01, 28.42, 22.22, 12.57%). Highe alues we e measu ed in he ui o he ´Hugin´ cul i a (31.12, 33.10, 30.48, 16.19%) and he highes sca enging ac i i y was obse ed in he ´Ne o´ cul i a (33.51, 37.30, 35.96, 19.22%) and he ´Viking´ cul i a (34.15, 41.46, 36.92, 19.81%). DISCUSSION Rega ding o 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 he cul i a a iabili y, which is qui e ypical o ui (Kopec, 1998). This a iabili y became e iden in case o TPC, TAC, ROS sca enging ac i i y and also lipid pe oxida ion ac i i y. 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 7.78 o 12.85 g o gallic acid/kg FM. These con en s co espond o he alues which we e ound ou in black chokebe ies by Zheng and Wang (2003). Mo eo e , some au ho s no iced high alues o phenols in black chokebe y ui , e en highe in compa ison wi h ou esul s (Ben enu i e al., 2004; Oszmianski and Wojdylo, 2005). Fo mos ui species lowe con en s a e ypical. Fo example, in apples he common con en is be ween 0.6 o 2.1 g o gallic acid/kg FM (V ho sek e al., 2004) o in plums i is 2.2 o 5.0 g o gallic acid/kg FM (Rop e al., 2009). An ioxidan capaci y was de e mined by he DPPH es and i was he highes in he cul i a ´Viking´ wi h he alue o 15.96 g o AAE/kg FM. In o he ui species, including apples (Hube and Rupasinghe, 2009), small be ies and s one ui (Chen e al., 2006), he alues o an ioxidan capaci y a e mos ly much lowe . Fo example, in che ies he e a e alues o up o 0.9 g o AAE/kg FM (Usenik e al., 2008) and in plums he alues can each up o 6 g o AAE/kg FM (Rop e al., 2009). As a as s a is ical e alua ion o he esul s is conce ned, he highes alues o co ela ion coe icien be ween an ioxidan capaci y and he o al amoun o phenolic subs ances we e ob ained (in case o he DPPH es 2 = 0.948, y = 1.5194x – 2.9191). Many au ho s no ice a high co ela ion be ween TPC and an ioxidan capaci y in ui (Thompson and Chao analiki , 2003; E cisli and O han, 2008). In e es ing esul s we e ecei ed in case o he con en s o pa icula phenols. While in case o cyanidin-3-glucoside he di e ences in i s con en we e sligh among he cul i a s (Table 2), in o he phenols he di e ences we e s a is ically signi ican . The highes con en s we e demons a ed in case o chlo ogenic acid ( om 1131.15 o 1960.72 mg/kg FM). This acid is an impo an phenol ound in black chokebe y ui (Skupien and Oszmianski, 2007). Zheng and Wang (2003) d aw a en ion o he ac ha chlo ogenic acid is he mos impo an an ioxidan in black chokebe y, which was also con i med in ou measu emen . The me hanolic ex ac s (25%) o black chokebe y ui showed mode a e inhibi o y abili y on hyd oxyl adical (34.15%), ni ic oxide (41.46%), supe oxide anion (36.92%) and lipid pe oxida ion (19.81%) – he cul i a ´Viking´. ROS a e known o cause aging, cance and many o he mal-e ec s on he human body and implica ed in pa hogenesis o se e al diseases (Sumanon e al., 2004). In his s udy, he black chokebe y ui ex ac was e alua ed o high abili y o sca enge hyd oxyl adical using he deoxy ibose deg ada ion assay. The cul i a s ´Ne o´ and ´Viking´ gene ally seemed o ha e sca enging ac i i ies o ROS including ni ic oxide. Sca enging ac i i y o supe oxide anion in he ex ac s o pa icula cul i a s was also demons a ed in he xan hine/xan hine oxidase sys em. Using ex ac s o black chokebe y ui was mo e e ec i e han in o he ui species, e.g. mulbe y (Bae and Suh, 2007), apples (Ma ei e al., 2007) o ui o Rop e al. 2435 1 2 3 4 5 6 7 Figu e 1. Ch oma og am ep esen ing he peaks o phenolics – he ´Viking´ cul i a ( he peak 2 = cyanidin-3-a abinoside, he peak 4 = cyanidin-3-galac oside, he peak 5 = chlo ogenic acid, he peak 6 = neochlo ogenic acid; he peak 7 = (-) epica echin. Table 3. Sca enging e ec o 25% black chokebe y ui me hanol ex ac s (pe cen age o inhibi ion). Cul i a Hyd oxyl adical Ni ic oxide Supe oxide anion Lipid pe oxida ion A on 25.01 ± 0.89a 28.42 ± 0.25a 22.22 ± 0.29a 12.57 ± 0.30a Fe ödi 22.08 ± 0.58b 27.59 ± 0.29b 21.24 ± 0.37b 12.05 ± 0.24a Hugin 31.12 ± 0.68c 33.10 ± 0.31c 30.48 ± 0.40c 16.19 ± 0.21b Ne o 33.51 ± 0.67d 37.30 ± 0.28d 35.96 ± 0.36d 19.22 ± 0.20c Viking 34.15 ± 0.51d 41.46 ± 0.31e 36.92 ± 0.38e 19.81 ± 0.26c 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. 2436 J. Med. Plan . Res. Table 2. The con en o phenolics (mg/kg FM). Cul i a cyanidin-3 a abinoside cyanidin-3-galac oside (-)epica echin chlo ogenic acid neochlo ogenic acid A on 941.82 ± 18.25a 1010.80 ± 34.25a 467.35 ± 4.56a 1254.20 ± 51.18a 883.31 ± 26.11a Fe ödi 972.90 ± 24.56a 1074.41 ± 31.50a 560.22 ± 9.74b 1131.15 ± 24.36b 845.04 ± 19.52a Hugin 1015.04 ± 29.01b 1071.91 ± 39.83a 647.08 ± 6.09c 1845.19 ± 21.12c 965.81 ± 27.41b Ne o 1420.17 ± 24.12c 1181.15 ± 28.20b 862.50 ± 8.70d 1931.45 ± 40.01d 1057.00 ± 25.30c Viking 1553.29 ± 35.44d 1203.56 ± 31.31b 844.44 ± 10.10d 1960.72 ± 37.11d 1156.59 ± 28.02d 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. P unus species (Jung e al., 2002). 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 (Wang e al., 2009). 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 and cell damage (Wang e al., 2008). The highes alues o lipid pe oxida ion ac i i ies we e obse ed in he cul i a s ´Ne o´ and ´Viking´, which con i ms biological e iciency o hese cul i a s in compa- ison wi h o he cul i a s e alua ed in his s udy. Conclusion In ou measu emen , e y high con en s o polyphenolic subs ances and o al an ioxidan capaci y we e obse ed in he black chokebe y ui . Fu he mo e, he e was a high co ela ion be ween polyphenolic subs ances and an ioxidan capaci y. In addi ion, high e iciency became e iden in eac i e oxygen species and lipid pe oxida ion sca enging ac i i ies. F om his poin o iew, he ui o black chokebe y ha e unique p ope ies in compa ison wi h o he ui species. They can become a aluable sou ce o nu i- ionally impo an subs ances in 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. 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