Influence of hot air drying on phenolic compounds and antioxidant capacity of blueberry (Vaccinium myrtillus) fruit and leaf

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1 Journl of Applie Botny n Foo Qulity 90, (2017), DOI: /JABFQ University of Bnırm Onyei Eylül, Bnırm Votionl Shool, Deprtment of Foo Proessing, Blıkesir, Turkey 2 University of Ulug, Agriulturl Fulty, Deprtment of Foo Engineering, Burs, Turkey 3 Kurtsn Phrmeutils In., Bnırm, Blikesir, Turkey 4 University of Blikesir, Engineering n Arhitehture Fulty, Deprtment Foo Engineering, Blikesir, Turkey Influene of hot ir rying on phenoli ompouns n ntioxint pity of lueerry (Vinium myrtillus) fruit n lef Nurn Değirmenioğlu 1*, Ozn Gürüz 2, Göze Erem Krtepe 3, Reyhn Irkin 4 (Reeive Otoer 20, 2016) * Corresponing uthor Summry The present stuy ws unertken to ssess the effets of hot ir rying on phenoli ompositions, totl phenoli (TP) ontent, totl nthoynin (TA) ontent, s well s ntioxint pities of methnol extrts from lueerry (Vinium myrtillus) fruit n lef introue in Kpığ region of Turkey limte onitions. A totl of twenty-two phenoli stnrs were sreene y HPLC, totl phenols were mesure y spetrophotometri methos, ntioxint pity ws etermine using DPPH, CUPRAC, ABTS, n FRAP ssys in the lueerry fruit n lef extrts. Anlysis y HPLC revele tht fruit extrts hve ifferent phenoli profiles ue to rying proess n ontin syringi i, myrietin, nringin, (-)-epitehin, n mlviine-3-o-gluosie hlorie s the min ompouns. Lef extrts h higher resvertrol onentrtions thn fruit extrts. The TP n TA ontents grully inrese when the lueerry fruits were rie uner hot ir onition. The fresh n rie lueerry fruit n lef extrts showe similr ntioxint pity vlues. Signifint reltionships etween ntioxint pity n TP were foun. Keywors V. myrtillus; lueerry; hot it rying; fruit; lef; phenolis; ntioxint pity Introution Blueerry, perennil shru of the genus Vinium, fmily Eriee, eme well known roun the worl ue to high levels of phenoli ompouns (Ehlenfelt n Prıor, 2001; Prıor et l., 2001; Kim et l., 2010; Routry et l., 2014). These ompouns hve een reporte to hve numerous vlule helth enefits inluing super ntioxint, nti-hypertensive, nti-ieti, nti-leukemi, nti-oesity, nti-inflmmtory, n nti-miroil tivity, s well s neurotive properties, to protet ginst ner n stroke (Ehlenfelt n Prıor, 2001; Deng et l., 2014; Li et l., 2013). Blueerries re onsiere to e one of the rihest soures of phenoli ompouns n ntioxint phytohemils mong fruits n vegetles, n they ontin signifint levels of nthoynins, flvonols, flvonons, pronthoyniins, n phenoli is (Cstrejón et l., 2008; Wng et l., 2012). Ftors tht hve n impt on the totl phenoli ontent, totl nthoynins, n the ntioxint pity of lueerries fruit n leves, inlue geneti ifferenes, the ultivr type, growing lotion n seson, gronomi ftors, the egree of mturity t hrvest, n posthrvest storge onitions (Ehlenfelt n Prıor, 2001; Deng et l., 2014). Drying, s preservtion metho, is very importnt spet of foo proessing. The min funtions of rying re lowering the wter tivity, inhiiting the growth of miroorgnisms, eresing hemil retions, extening shelf life, llowing for room temperture storge, reuing trnsporttion osts with regr to refrigertion, n lso enhning visul n tste of erels, onfetions, n ke goos. Severl rying tehniques suh s sun rying, onvetion oven rying, freeze rying, mirowve rying et., hve een employe in n effort to hieve high qulity rie lueerries (Meji-Mez et l., 2008; Hmrouni-Sellmi et l., 2013). The seletion of rying methos to e use is epenent on the use of the en prout, eonomi viility, vilility of resoures, n omposition of the iomteril (Routry et l., 2014). Convetive hot ir rying is tritionl, low ost tehnique tht is wiely use to lower the wter ontent of fresh prouts t present, nevertheless it requires reltively long times n high tempertures, uses egrtion of importnt nutrients, hs shrunken n toughene rie prouts with notiele rowning, n llows for little rehyrtion ility (Sellppn et l., 2002). The im of this stuy ws to etermine the influene of lotion, prt of plnt (fruit or lef), n oven rying on the phenoli ompouns, totl phenol n nthoynin ontents, n ntioxint pities of lueerries (V. myrtillus). Mteril n methos Smple olletion n preprtion Blueerry leves n fruit grown t three ifferent lotions of Erek (se level, Blikesir, Turkey) n Kpığ (ltitues of 650 m, Blıkesir, Turkey) regions were rnomly hn-pike uring Otoer n Novemer, 2014, n trnsporte to the lortory within the sme y. Upon rrivl, the fruit n leves were hn selete n seprte into two lots of equl weight. One lot of fruit n leves were store seprtely fresh, n the other lots were firstly rie t 50 C for 2 h in n ir oven type FN 055 (NUVE, İstnul, Turkey) then oole. The two lots were then seprtely vuum pkge (VC 999/K12NA pking mhine, Verpkungssysteme AG, Herisu, Switzerln) in FMXBK polymie-polyethylene film (PO 2 =15 m 3 / m 2 /24 h t 23 C n 75% reltive humiity; Flexopk S.A. Plstis Inustry, Koropi, Greee) n store t -20 C (SF 312, Direi, Tokyo, Jpn) until further nlysis. Extrts of V. myrtillus fruit n lef were prepre oring to the metho of esrie y Ehlenfelt n Prior (2001) n Prior et l. (2001), with some moifitions. Fresh n rie fruit n leve smples (2 g) were seprtely extrte twie with 20 ml of methnol:formi i (99.5/0.5, v/v, for fruit), n etone:formi i (99.5/0.5, v/v, for lef) mixture in n ultrsoni th t room temperture (20 C) for 15 min. Extrts then were seprtely entrifuge t 3500 rpm for 10 min t 4 C in entrifuge (Sigm 3K30, UK). The superntnts were omine, fter removl of methnol n etone with rotry evportor (Heiolph Lorot 4001, Germny) uner vuum onitions t 40 C, the resiul extrts were sujete to liqui-liqui prtition with methnol:formi i (99.5/0.5, v/v, for fruit) n etone:formi i (99.5/0.5, v/v, for leves), respetively, filtere through nylon filter memrne (Sigm Z290793, pore size 0.45 μm, im. 47 mm), trnsferre to vils, n store -20 C until further nlysis.

2 116 N. Değirmenioğlu, O. Gürüz, G.E. Krtepe, R. Irkin Chemils Phenoli stnrs were otine from Fluk (St. Louis, MO, USA) (glli i; CAS: , feruli i; CAS: , resvertrol; CAS: , (+)-tehin; CAS: , (-)-epitehin; CAS: , myrietin; CAS: , kempferol; CAS: , (-)-epigllotehin; CAS: , Sigm (St. Louis, MO, USA) (queretin; CAS: , ffei i; CAS: , syringi i; CAS: , p-oumri i; CAS: , nringin; CAS: , hesperiin; CAS: , neohesperiin; CAS: , rutin hyrte; CAS: , yniin-3-oglyosie hlorie; CAS: , mlviine-3-o-glyosie hlorie; CAS: ), Alrih (St. Louis, MO, USA) (vnilli i; CAS: , trns feruli i; CAS: , 3-hyroxy-4- metoxy-innmi i; CAS: ), HWI Anlytik GmH (Ruelzheim, Germny) (hlorogeni i; CAS: ). Clirtion urves were me y iluting stok stnrs in methnol. Determintion of phenoli omposition using HPLC Phenoli ompositions were nlyse oring to previously reporte metho with moifitions in HPLC elution onitions (Sellppn et l., 2002). The phenoli extrts, phenoli stnrs n lso ll solvents were filtere through nylon filter memrne (Sigm Z290793, pore size 0.45 μm, im. 47 mm) prior to HPLC nlysis n then nlyse in HPLC hromtogrphy system (Shimzu Clss VP V.6.14 SP1, USA) equippe with Shimzu Dioe Arry Detetor (SPD-M 10A), VP n reverse-phse C18 olumn (Zorx Elipse XDB, Agilent, 4.6 mm, 150 mm, 5 μm). The temperture of the olumn oven ws set t 40 C. The wvelengths use for the quntifition of phenoli ompouns y the etetor were: 280 nm for syringi i, glli i, (+)-tehin, neohesperiin, ffei i, hesperiin, (-)-epigllotehin, (-)-epitehin, nringin, vnilli i; 320 nm for trns-feruli i, hlorogeni i, 3-hyroxy-4-metoxy-innmi i, resvertrol, p-oumri i, feruli i; 360 nm for myrietin, rutin hyrte, kempferol, queretin; n 520 nm for yniin-3-o-glyosie hlorie, mlviine-3-oglyosie hlorie. A grient elution ws employe with moil phse onsisting of methnol:wter:formi i (3.5/96.4/0.1, v/v/v, solvent A) n etonitril:formi i (98/2, v/v, solvent B) s follows: the omposition of B ws inrese from 0.5% to 7.5% fter 31 min, inrese to 10% for 9 min, n inrese to 14% for 5 min, inrese to 18% for 5 min, inrese to 30% for 10 min, inrese to 45% for 5 min, n inrese to 60% for 5 min. The omposition ws erese to 40% for 5 min. The injetion volume ws 20 μl, the flow rte ws 0.7 ml/min t room temperture, the urtion of single run ws 75 min. All phenoli is were quntifie using n externl stnr. The totl phenoli extrts n stnr ompouns were nlyze uner the sme nlysis onitions n 10 min equilirium time ws llowe etween injetions. All stnr n smple solutions were injete in triplite. Determintion of totl phenoli (TP) ontent The totl phenoli (TP) ontents of fresh n rie lueerry fruit n leve extrts were mesure y the Folin-Ciolteu metho esrie y Singleton et l. (1999), with some moifitions. Briefly, n liquot (0.5 ml) of ppropritely ilute extrts, or stnr solutions of glli i, 1.5 ml of oule istille wter n 2.5 ml Folin-Ciolteu regent, were mixe within volumetri flsks t room temperture. After 10 min, 0.25 ml of 7.5% soium ronte (1:3 ilute with oule istille wter) solution (m/v) ws e n mixe thoroughly. The sorne of the solution ws mesure using spetrophotometer (UVMesys Optizen 3220) t 750 nm fter 30 min in the rk t room temperture. Methnol ws use s the lnk n glli i ws use for lirtion of the stnr urve (0-500 mg/l). The results were expresse s mg of glli i equivlents (GAE) per kg. Eh extrt ws mesure in triplite. Determintion of totl nthoynin (TA) ontent The totl nthoynin ontent of extrts otine from lueerry fruit n leves were etermine y mens of the ph-ifferentil metho s esrie y Sellppn et l. (2002). The sorne ws mesure using spetrophotometer (UVMesys Optizen 3220) t 700 nm n t the wvelength of mximum sorption (520 nm) ginst lnk n lulte s: A = (A 520 A 700 ) ph (A 520 A 700 ) ph 4.5 Monomeri nthoynin pigment onentrtion of extrts ws lulte s yniin-3-gluosie equivlent n eh extrt ws mesure in triplite. Monomeri nthoynin pigment (mg/l) = A MW DF 1000 (ε 1) where A = sorne, MW = moleulr weight (449.2), DF = ilution ftor, ε = molr sortivity (26900). The finl onentrtion of totl nthoynins (mg/kg) ws lulte se on totl volume of extrt n weight of smple. Determintion of ntioxint pity y upri ion reuing ntioxint pity (CUPRAC) ssy Determintion of CUPRAC ws onute oring to the metho y Apk et l. (2007). One ml 10 mmol/l CuCl 2, 1 ml 7.5 mmol/l neouproine, 1 ml 1 M NH 4 A, ml extrt, n (4- ) ml H 2 O were mixe. The tues were stoppe n fter 30 min the finl sorne ws reore using spetrophotometer (UVMesys Optizen 3220) t 450 nm ginst regent lnk. A stnr urve ws prepre using ifferent onentrtions of Trolox. The lultions of the ntioxint pity of phenoli ntioxints were expresse s μmol of Trolox equivlent (TE) per grm. Eh extrt ws mesure in triplite. Determintion of ntioxint pity y DPPH (2,2-iphenyl-2- pirylhyrzyl) free ril ssy The free ril svenging pity of the lueerry fruit n leve extrts were etermine y olorimetri metho esrie y Brn-Willims et l. (1995). In rief, the ppropritely ilute extrts ( ml), methnol (4- ml), n DPPH solution (3.9 ml, 50 μm) in methnol were inute in wter th t 37 C for 30 min. After inution, the sorne ws mesure t 515 nm with spetrophotometer (UVMesys Optizen 3220). The results were lulte ginst methnol without DPPH n ompre to ifferent onentrtion of Trolox stnr urve. Eh extrt ws mesure in triplite. DPPH vlues, erive triplite nlyses, were expresse s μmol of Trolox equivlent (TE) per grm n were lulte s follows: DPPH ril svenging pity (%) = (1-[A smple /A ontrol ]) 100 Determintion of ntioxint pity y ABTS [2,2-zinois(3- ethylenzothizoline-6-sulphoni i)] ssy The ABTS metho is se on the etivtion of the ntioxint ril tion ABTS +. The ABTS metho ws performe s esrie y Re et l. (1999). ABTS ril tion (ABTS + ) ws proue y reting 7 mm ABTS solution with 2.45 mm potssium persulfte (K 2 S 2 O 8 ) queous solution n llowing the mixture to stn in the rk t room temperture for h efore use. Different onentrtions of fruit n lef extrts were mixe with 1 ml of ilute ABTS + solution n the reution of ABTS + ril ws mesure y the erese in sorne t 734 nm fter 6 min y

3 Biotive ompouns in fruits n leves of lueerry 117 using the spetrophotometer UVMesys Optizen To evelop stnr urve, stnr Trolox solution ws ilute with ethnol n e to 1 ml of the ilute ABTS + solution. The ontrols ontine the extrtion solvent inste of the test smples. Eh extrt ws mesure in triplite. The svenging pity of ABTS free ril ws lulte s: ABTS ril svenging pity (%) = (1-[A smple /A ontrol ]) 100 Determintion of ntioxint pity y FRAP ssy The FRAP ssy ws onute oring to Benzie n Strin (1996). This metho is se on n inrese of the sorne t 593 nm ue to the formtion of tripyriyl-s-trizine omplexes with Fe 2+ [TPTZ-Fe(II)] in the presene of reutive gent. The FRAP regent ws prepre y mixing TPTZ solution (10 mmol/l) in hyrohlori i (40 mmol/l) n FeCl 3 solution (20 mmol/l) mixe with 25 ml of ette uffer (0.3 mol/l, ph=3.6). An ppropritely ilute smple extrt ( μl) n FRAP regent (1- ml) were e n, the mixture n extrtion or solvent for the regent lnk were inute t 37 C for 30 min. At the en of inution, sorne ws immeitely mesure using spetrophotometer (Perkin Elmer UV/VIS Lm35) t 595 nm. Solutions of Trolox issolve in extrtion solvent, rnging from μmol/l were use for preprtion of lirtion urve. FRAP vlues, erive from triplite nlyses, n were expresse s μmol of Trolox equivlent (TE) per grm. Eh extrt ws mesure in triplite. Sttistil nlysis Sttistil ifferenes etween the t sets were etermine y two-wy Anlysis of vrine (ANOVA) using the SPSS sttistil pkge (SPSS 16.0, Chigo, IL). Differenes etween tretments tht re esrie susequently s eing signifint, were etermine t lest p<0.05. The lest signifint ifferene (LSD) test ws use to etermine ifferenes etween mens. Results n isussion Phenoli ompositions The t set for the ontents of phenoli is, flvonols, flvnones, monomeri of flvn-3-ol erivtives, nthoynins, n the stilene in the V. myrtillus fresh n rie fruit n lef extrts re given in T These ompouns n t s ntioxints n my e importnt omponents of funtionl foos. The ominnt phenoli i ws syringi i in fresh fruit extrts grown in Erek n Kpığ regions ( mg/kg FW, mg/kg FW, respetively), n their ontent ws espeilly high (p<0.05) in rie fruits. In fresh n rie V. myrtillus lef extrts, syringi, p-oumri, glli n vnilli is were the most unnt phenoli is. Some reserh hs reporte tht hlorogeni i, referre to s 5-O-ffeoylquini i (5-CQA), is onsiere mjor olourless phenoli i in lueerry fruit n lef (Hrris et l., 2007; Kim et l., 2010), n more reily ville soures of 5-CQA, even ompre to green offee ens (Kim et l., 2010). Prior et l. (2001) foun the level of hlorogeni i in lueerries to e mg/g of fresh fruit, while Hrris et l. (2007) etete this ompoun 30 times more onentrte in the lef extrt thn in fruit. Nevertheless, in this stuy high hlorogeni i levels in the fruit n lef extrts were not etermine. On the other sie, vnilli i, 3-hyroxy-4-metoxy innmi i, n feruli i were otine in fresh fruit extrts, wheres in rie fruit extrts, higher levels of vnilli i, glli i, p-oumri i, feruli i, n 3-hyroxy-4-metoxy innmi i were etermine. Aprt from hlorogeni i, the phenoli is hve een foun to e present in smller onentrtions, suh s ffei, p-oumri n feruli i (Sellppn et l., 2002). In ition, other phenols tht my e foun inlue glli, p-hyroxyenzoi, m-hyroxyenzoi, ellgi, vnilli, prototehui, gentisi, syringi, sinpi n sliyli is, n tehin, epitehin, myrietin, n kempferol (Sellppn et l., 2002; Hrris et l., 2007). In our smples otine from Erek n Kpığ, we foun some of these omponents, n most phenoli ompouns etete in this stuy were onsistent with previous reports on lueerry fruit n leves from ifferent lotions in the worl. It is elieve tht the signifint qulittive n quntittive ifferenes (p<0.05) of phenoli ompouns profiles whih hve een onfirme in lueerries, re ue to vritions in genotypes, lotions, ultivtion onitions, inrese mturity, ifferent prts of plnts exmine, stresses, orgnilly grown, extrtion methos, n ll n hve vrying effets on the level of totl nthoynins, totl phenolis n ntioxint pity (Xioyong n Luming, 2014). Also rying onitions n use ifferenes. Some reent stuies hve een omplishe on the ontent of lueerry n ilerry ntive flvonols (Sellppn et l., 2002; Hrris et l., 2007; Moze et l., 2011; Vrhovsek et l., 2012). In this stuy, fresh V. myrtillus fruit extrts ontine low flvonols ontents ( mg/kg FW), wheres rie fruit n lef extrts ontine reltively high flvonols. While myrietin ws foun to e the min flvonol ompoun in the leves, kempferol n queretin were lso etete, in greement with other stuies. Previous reserh lso etermine tht the green leves of lueerry ontine muh lrger mount of flvonois (queretin n kempferol) n hyroxyinnmi i (p-oumri n ffei i) thn fruits (Riihinen et l., 2008). Nevertheless, in this stuy the onentrtions of myrietin were higher in the fresh leves extrts thn fresh fruit extrts (p<0.05). Also in the rie fruit extrts, the onentrtions of flvonols were onsierly higher thn in the rie lef extrts. As mentione y Jkol et l. (2004) n Oszmiński et l. (2011), p-oumri i is the preursor of flvonois, n the inrese in p-oumri i onentrtion in the leves, growing uner high solr rition, n lso reflet the overll tivtion of flvonoi iosynthesis. T. 1-4 shows tht ll the flvonol ompouns h some signifint (p<0.05) level of vriility ue to ltitue, n ue to the rying proess. Rutin hyrte, whih hs een reporte in high mounts in ukwhet (Moze et l., 2011), ws foun in oth regions of V. myrtillus fruits uner investigtion, the rie fruit extrts ontining more rutin hyrte thn fresh fruit extrts n lso ompre to the fresh n rie lef extrts. Moze et l. (2011) first etete rutin in ilerry n lueerry smples (0.2 n 3.1 mg/100g FW, respetively), n lso rutin (queretin-3-o-rutinosie) ws the mjor phenoli ompouns in leves of riteye lueerry ultivte in Jpn (Li et l., 2013). Hrris et l. (2007) etermine this ompoun s 3.10 mg/100 g in V. oryoum L. fruits, wheres our results were higher thn their results. Bioflvonois like rutin n nringin hve een proven to e effiious ntioxints n re wiely istriute in fruits n vegetles. Rutin elongs to the lss of flvonols n nringin elongs to flvnones. It is well note n proven in severl stuies, tht flvonols re very tive in onveying therpeuti enefit ompre to flvonones (Akoni et l., 2011). The flvonones (nringin, hesperiin, n neohesperiin) were etete in stuie V. myrtillus extrts. Nringin is one of the most unnt flvnone in fresh n rie fruit n lef extrts in this stuy. It ws lso oserve tht nringin levels were greter in smples hrveste from high ltitue ompre to those smples originting from res t se level. Some reports hve inite tht rying methos n ffet phenoli ontents n the ntioxint pity of plnt mterils ue to rying time/temperture, light intensity, pkging, n storge time et. (Lu n Luthri, 2014). The three flvnols were foun in oth regions of V. myrtillus, the rie fruit extrts eing etter soure of (+)-tehin, (-)-epite-

4 118 N. Değirmenioğlu, O. Gürüz, G.E. Krtepe, R. Irkin T. 1: Phenoli ompouns onentrtions of fresh V. myrtillus fruits grown in Erek ve Kpığ regions (mg/kg) Phenoli ompouns E1FRF * E2FRF E3FRF K1FRF K2FRF K3FRF Glli i ± 2.12 C** ± 4.58 A ± 3.14 B ± 1.13 Dk 8.10 ± 1.10 Dl 0.87 ± 0.12 Em Vnilli i ± A ± 3.25 C ± 5.12 B ± 1.95 El 9.25 ± 1.63 Fm ± 1.98 Dk Cffei i 5.46 ± 1.23 A 4.84 ± 1.10 B 5.35 ± 1.00 A 1.87 ±0.12 Ck 1.86 ± 0.24 Ck 1.73 ± 0.23 Dl Chlorogeni i 1.25 ± 0.25 E 4.37 ± 0.85 A 1.71 ± 0.26 C 1.42 ± 0.10 Dl 1.50 ± 0.27 CDl 2.71 ± 0.45 Bk Syringi i ± 3.45 B ± 4.85 F ± 1.26 E ± 6.52 Cl ±6.54 Ak ± 3.21 Dm p-coumri i 7.63 ± 1.48 C 1.81 ± 0.12 E 4.04 ± 1.85 D ± 2.13 Bl ± 2.31 Ak 8.68 ± 1.00 Cm Feruli i ± 2.69 D 6.06 ± 0.95 E 4.13 ± 0.74 F ± 2.31 Bl ± 1.98 Ak ± 1.14 Cm Trns-feruli i 5.99 ± 1.78 D 1.89 ± 0.14 E 5.45 ± 1.23 D 8.49 ± 2.85 Cm ± 2.14 Bl ± 1.56 Ak 3-hyroxy-4-methoxy ± 4.65 D ± 4.65 C ± 2.32 E ± 3.12 BCm ± 6.54 Ak ± 3.24 Bl innmi i Myrietin ± 2.98 D ± 3.12 D ± 2.96 E ± 2.96 Cm ± 3.45 Ak ± 3.59 Bl Queretin 1.62 ± 0.12 E 4.27 ± 1.00 B 6.93 ± 1.85 A 2.48 ± 0.62 Dl 3.71 ± 0.56 Ck 2.24 ± 0.46 Dl Kempferol 1.72 ± 0.45 AB 0.94 ± 0.16 D 1.82 ± 0.15 A 0.79 ± 0.10 Em 1.01 ± 0.15 Dl 1.67 ± 0.37 Bk Rutin hyrte 3.84 ± 1.10 D 1.31 ± 0.25 F 2.07 ± 0.23 E ± 1.10 Bl ± 1.47 Ak 7.64 ± 0.96 Cm Nringin 5.50 ± 1.14 D ± 3.74 B ± 3.41 A 4.42 ± 0.65 Em 6.97 ± 0.84 Ck 6.13 ± 0.45 Dl Hesperiin 7.07 ± 1.56 C ± 2.85 B ± 2.16 A 2.40 ± 0.13 El 4.93 ± 0.16 Dk 2.35 ± 0.78 El Neohesperiin 2.50 ± 0.45 B 8.38 ± 2.14 A 8.80 ± 1.62 A 1.83 ± 0.14 CDl 1.92 ± 0.11 Ck 1.73 ± 0.15 Cm (+)-Ctehin 2.72 ± 0.41 D ± 2.48 A 9.45 ± 1.64 B 6.41 ± 0.64 Ck 1.36 ± 0.12 Em 2.63 ± 0.43 Dl (-)-Epitehin ± 3.41 D ± 1.96 E 8.63 ± 1.18 F ± 2.63 Bl ± 4.85 Ak ± 2.45 Cm (-)-Epigllotehin 6.66 ± 1.58 D ± 3.11 B ± 3.98 A ± 1.84 Cl ± 2.41 Bk ± 1.47 Cm Cyniin-3-O-gluosie ± C ± 4.23 E ± 4.85 C ± 5.23 Bl ± 6.52 Ak ± 6.21 Dm hlorie Mlviine-3-O-gluosie ± C ± 6.52 F ± 6.12 E ± 8.95 Bl ± 9.96 Ak ± 5.27 Dm hlorie Resvertrol 1.01 ± 0.05 A 0.87 ± 0.05 B 0.78 ± 0.05 B 0.70 ± 0.06 Bm 0.84 ± 0.06 Bl 1.00 ± 0.00 Ak * E: Erek, K: Kpığ, FR: Fresh, F: Fruit, 1-3: Coes of smples ollete from ifferent regions, ** Men vlues (mg/kg)±stnr evition (N=3 2) with ifferent pitl letters (A-F) in the sme row re signifintly ifferent (p<0.05) oring to ollete from ifferent region t fresh fruit. Men vlues±stnr evition (N=3 2) with ifferent lowerse (-. k-m) in the sme row re signifintly ifferent (p<0.05) oring to ollete from the sme region t fresh fruit. T. 2: Phenoli ompouns onentrtions of rie V. myrtillus fruits grown in Erek ve Kpığ regions (mg/kg (mg/kg) Phenoli ompouns E1DRF * E2DRF E3DRF K1DRF K2DRF K3DRF Glli i ± 4.78 C** ± 1.78 E ± 5.41 B ± 2.18 Bl ± 1.10 Dm ± 3.25 Ak Vnilli i ± 3.65 B ± 3.65 A ± 3.29 C ± 2.87 Em ± 2.85 Ck ± 4.58 Dl Cffei i 3.92 ± 1.85 D 2.19 ± 0.18 E 6.99 ± 1.23 C 3.80 ± 0.51 Dl ± 1.47 Ak 31.88± 1.95 Bm Chlorogeni i 1.19 ± 0.14 E 0.94 ± 0.02 F 1.66 ± 0.45 D 2.11 ± 0.11 Cm 8.04 ± 1.45 Ak 2.86 ± 0.10 Bl Syringi i ± D ± 1.85 F ± C ± Bl ± Ak ± 3.85 Em p-coumri i ± 2.14 E 9.09 ± 1.63 F ± 3.45 D ± 3.61 Bl ± 3.25 Ak ± 4.45 Cm Feruli i ± 3.16 D 7.49 ± 1.74 E ± 2.89 C ± 3.54 Ak ± 1.85 Bl ± 6.18 Bl Trns-feruli i 1.54 ± 0.14 E 1.06 ± 0.16 E 2.52 ± 0.85 D ± 4.52 Ak ± 3.85 Cm ± 2.85 Bl 3-hyroxy-4-methoxy ± 2.96 C ± 2.65 E ± 2.84 A ± 3.21 Bl ± 3.84 Ak ± 4.78 Dm innmi i Myrietin ± 4.32 D ± 2.85 E ± 9.12 A ± 9.85 Ak ± 5.45 Bl ± 6.59 Cm Queretin 2.94 ± 0.56 D 1.13 ± 0.13 E 4.67 ± 1.12 C 9.67 ± 1.25 Bl ± 1.84 Ak 4.36 ± 0.19 Cm Kempferol 8.87 ± 1.12 D 3.82 ± 0.41 E ± 1.45 C ± 7.84 Ak ± 2.14 Cm ± 1.18 Bl Rutin hyrte ± 1.45 E 8.53 ± 0.45 F ± 3.12 C ± 6.10 Ak ± 6.95 Bl ± 2.14 Dm Nringin ± 2.34 D ± 2.95 D ± 4.59 C ± 6.25 Ak ± 6.54 ABkl ± 5.61 Bl Hesperiin 2.69 ± 0.16 D 2.30 ± 0.16 D 3.55 ± 0.48 C ± 1.42 Ak ± 2.14 Ak 8.46 ± 1.26 Bl Neohesperiin 1.23 ± 0.14 DE 0.94 ± 0.05 E 1.66 ± 0.12 D 3.58 ± 0.95 Bl 2.97 ± 0.23 Cm 6.14 ± 1.14 Ak (+)-Ctehin 1.70 ± 0.19 F 5.63 ± 0.96 E ± 0.41 B 7.21 ± 1.58 Dm ± 1.45 Cl ± 4.40 Ak (-)-Epitehin ± 2.64 D ± 2.46 E ± 9.65 C ± Bl ± Ak ± 9.98 Cm (-)-Epigllotehin ± 1.97 E ± 1.85 F ± 5.12 D ± 2.52 Bl ± 6.95 Ak ± 3.48 Cm Cyniin-3-O-gluosie ± 6.45 D ± 3.47 E ± B ± Cl ± Ak ± 3.87 Fm hlorie Mlviine-3-O-gluosie ± 5.23 D ± 5.42 E ± A ± Cl ± Bk ± Em hlorie Resvertrol 1.07 ± 0.12 DE 0.97 ± 0.04 E 1.12 ± 0.14 D 1.37 ± 0.10 Cm 2.18 ± 0.12 Bl 2.58 ± 0.13 Ak * E: Erek, K. Kpığ, DR: Drie, F: Fruit, 1-3: Coes of smples ollete from ifferent regions, ** Men vlues (mg/kg)±stnr evition (N=3 2) with ifferent pitl letters (A-F) in the sme row re signifintly ifferent (p<0.05) oring to ollete from ifferent region t rie fruit. Men vlues±stnr evition (N=3 2) with ifferent lowerse (-. k-m) in the sme row re signifintly ifferent (p<0.05) oring to ollete from the sme region t rie fruit.

5 Biotive ompouns in fruits n leves of lueerry 119 T. 3: Phenoli ompouns onentrtions of fresh V. myrtillus leves grown in Erek ve Kpığ regions (mg/kg) Phenoli ompouns E1FRL * E2FRL E3FRL K1FRL K2FRL K3FRL Glli i ± 1.65 C** ± 2.14 B 6.53 ± 0.96 E ± 3.58 Ak ± 1.26 Cl 7.17 ± 1.12 Dm Vnilli i ± 6.85 B ± 6.51 C ± 3.48 E ± 2.14 Fm ± Ak ± 9.87 Dl Cffei i ± 9.78 A ± B ± 2.14 D ± 4.59 Ck ± 3.85 Dl ± 2.15 Em Chlorogeni i 9.07 ± 1.23 A 3.16 ± 0.29 B 3.41 ± 0.27 B 2.27 ± 0.52 CDl 2.20 ± 0.84 Dl 2.49 ± 0.48 Ck Syringi i ± 2.14 C ± A ± 3.56 D ± 3.65 El ±10.84 Bk ± 2.47 El p-coumri i ± 6.54 A ± 6.58 C ± 5.84 D ± 9.85 Bl ± Ak ± 3.65 Em Feruli i 5.51 ± 1.10 BC 5.23 ± 0.95 D 5.33 ± 1.10 CD 5.59 ± 1.05 BCl 5.68 ± 0.95 Bl 6.50 ± 0.98 Ak Trns-feruli i 9.98 ± 1.18 B ± 1.84 A 5.19 ± 0.56 C 4.48 ± 0.96 Dl 4.38 ± 0.28 Dl 5.27 ± 0.82 Ck 3-hyroxy-4-methoxy ± 3.87 B ± 3.58 F ± 3.98 C ± 4.12 Dl ± 6.54 Ak ± 6.14 Em innmi i Myrietin ± 6.41 B ± 4.50 D ± 6.52 C ± 9.23 Cl ± 8.59 Ak ± Dm Queretin 2.26 ± 0.25 D 0.99 ± 0.19 E 9.32 ± 1.12 C 9.97 ± 1.10 BCm ± 1.65 Ak ± 3.45 ABl Kempferol 2.56 ± 0.39 C 1.28 ± 0.25 F 9.91 ± 1.48 A 1.79 ± 0.27 Dl 1.38 ± 0.29 Em 8.78 ± 2.15 Bk Rutin hyrte 5.74 ± 0.48 D ± 1.17 A 9.81 ± 2.00 B 5.53 ± 0.52 Dl 2.46 ± 0.54 Em 8.95 ± 1.84 Ck Nringin ± 1.85 A 4.45 ± 0.98 D ± 2.58 B 5.10 ± 0.29 Dl ± 2.87 Ck ± 2.14 BCk Hesperiin ± 1.74 A 7.50 ± 1.25 C 9.56 ± 2.05 B 6.31 ± 0.84 Dl 5.89 ± 0.68 Em 7.73 ± 0.96 Ck Neohesperiin 2.81 ± 0.69 BC 2.68 ± 0.62 C 7.73 ± 2.14 A 2.36 ± 0.26 Dl 2.19 ± 0.39 Em 3.12 ± 0.25 Bk (+)-Ctehin ± 2.98 E ± 3.48 C ± 9.51 B ± 8.63 Ak ± 5.62 Dl ± 2.15 Fm (-)-Epitehin ± 2.45 B 7.06 ± 1.18 D 5.60 ± 1.10 E 2.55 ± 0.35 Fm ± 2.10 Cl ± 2.46 Ak (-)-Epigllotehin 9.84 ± 2.12 C ± 3.15 B ± 8.56 A 8.70 ± 0.95 Dk 7.23 ± 1.98 El 9.12 ± 1.10 CDk Cyniin-3-O-gluosie 1.51 ± 0.23 B 1.02 ± 0.20 C 0.96 ± 0.18 D 0.98 ± 0.27 CDl 0.97 ± 0.19 CDl ± 2.58 Ak hlorie Mlviine-3-O-gluosie 1.57 ± 0.15 B 1.10 ± 0.14 C 0.96 ± 0.12 D 1.05 ± 0.16 CDl 0.00 ± 0.00 Em ± 5.00 Ak hlorie Resvertrol 1.57 ± 0.10 E 4.11 ± 0.58 C 6.29 ± 1.10 A 3.91 ± 0.56 Dm 5.89 ± 0.84 ABk 5.31 ± 0.85 Bl * E: Erek, K: Kpığ, FR: Fresh, L: Lef, 1-3: Coes of smples ollete from ifferent regions, ** Men vlues (mg/kg)±stnr evition (N=3 2) with ifferent pitl letters (A-F) in the sme row re signifintly ifferent (p<0.05) oring to ollete from ifferent region t fresh lef. Men vlues±stnr evition (N=3 2) with ifferent lowerse (-. k-m) in the sme row re signifintly ifferent (p<0.05) oring to ollete from the sme region t fresh lef. T. 4: Phenoli ompouns onentrtions of rie V. myrtillus leves grown in Erek ve Kpığ regions (mg/kg) Phenoli ompouns E1DRL * E2DRL E3DRL K1DRL K2DRL K3DRL Glli i ± 6.98 C** ± 3.59 A ± 3.29 B ± 4.36 Bk ± 3.69 Em ± 9.62 Dl Vnilli i ± 5.89 B ± A ± 4.62 C ± 2.48 Dl ± Ck ± 2.98 Em Cffei i ± 3.21 A 5.28 ± 1.14 C ± 2.12 A 8.61 ± 1.10 Bk 4.65 ± 0.85 Dl 3.28 ± 0.68 Em Chlorogeni i 3.31 ± 0.45 B 3.23 ± 0.58 C 1.25 ± 0.26 E 4.14 ± 0.87 Ak 1.94 ± 0.34 Dm 3.25 ± 0.52 BCl Syringi i ± 5.47 A ± 6.47 D ± 3.29 B ± 6.54 Ck ± 3.85 El ± 5.20 Fm p-coumri i ± 2.85 B ± 2.19 D ± 1.23 C 8.51 ± 1.98 Em ± 2.18 Bl ± 2.16 Ak Feruli i 3.13 ± 0.95 E 3.93 ± 0.48 C 3.80 ± 0.48 CD 9.99 ± 2.57 Ak 3.50 ± 0.84 DEm 6.56 ± 1.05 Bl Trns-feruli i ± 2.14 D ± 2.43 DE ± 1.15 A ± 2.18 Bk ± 2.17 Cl ± 2.84 Em 3-hyroxy-4-methoxy ± 4.58 C ± 4.56 A ± 3.58 E ± 9.84 Bk ± 9.27 Dl ± 9.13 Fm innmi i Myrietin ± 4.32 D ± 5.42 A ± 4.51 D ± Bk ±12.17 Cl ± 7.68 Em Queretin 4.63 ± 0.47 C 2.07 ± 0.23 D 8.10 ± 1.18 B 4.27 ± 0.87 Cl 2.75 ± 0.96 Dm ± 2.34 Ak Kempferol 1.63 ± 0.28 D 3.37 ± 0.43 A 2.17 ± 0.18 B 2.20 ± 0.59 Bl 3.30 ± 0.95 Ak 2.07 ± 0.56 Bl Rutin hyrte ± 1.15 D ± 6.58 A ± 2.15 C ± 2.51 Bk ± 3.25 El 8.08 ± 1.02 Fm Nringin ± 5.21 B ± 5.53 A ± 6.41 C ± 3.26 El ± 8.26 Dk 1.14 ± 0.38 Fm Hesperiin ± 5.10 B ± 4.87 A ± 4.28 C ± 3.48 Dk 7.45 ± 2.14 Fm 9.74 ± 1.36 El Neohesperiin 6.52 ± 0.75 B 3.18 ± 0.59 C 6.06 ± 0.58 B 17.24± 2.47 Ak 2.63 ± 0.51 Cm ± 2.01 Al (+)-Ctehin ± 1.48 D ± 3.57 A ± 2.43 B ± 2.52 Cl 7.31 ± 2.18 Em ± 3.20 Bk (-)-Epitehin ± 3.47 B ± 2.15 D ± 1.64 D ± 9.52 Ak ± 6.20 Cm ± 4.02 Bl (-)-Epigllotehin ± 3.65 C ± 2.39 E ± 3.28 D ± 6.53 Bl ± 3.27 Dm ± Ak Cyniin-3-O-gluosie 0.00 ± 0.00 C 1.00 ± 0.06 AB 1.01 ± 0.17 AB 1.06 ± 0.26 Ak 1.01 ± 0.18 ABkl 0.95 ± 0.09 Bl hlorie Mlviine-3-O-gluosie 1.09 ± 0.18 AB 0.00 ± 0.00 C 1.14 ± 0.16 AB 1.20 ± 0.19 Ak 1.11 ± 0.24 ABkl 0.95 ± 0.12 Bl hlorie Resvertrol 3.32 ± 0.42 B 1.54 ± 0.28 C 1.50 ± 0.45 C 8.89 ± 2.21 Ak 8.55 ± 1.00 Ak 3.85 ± 0.45 Bl * E: Erek, K. Kpığ, DR: Drie. L: Lef, 1-3: Coes of smples ollete from ifferent regions, ** Men vlues (mg/kg)±stnr evition (N=3 2) with ifferent pitl letters (A-F) in the sme row re signifintly ifferent (p<0.05) oring to ollete from ifferent region t rie lef. Men vlues±stnr evition (N=3 2) with ifferent lowerse (-. k-m) in the sme row re signifintly ifferent (p<0.05) oring to ollete from the sme region t rie lef.

6 120 N. Değirmenioğlu, O. Gürüz, G.E. Krtepe, R. Irkin hin, n (-)-epigllotehin ompre to fresh fruit n lef n lso rie lef extrts. The flvnol ontents in fruit n lef extrts were etermine to e inresing or eresing epening on the ltitue, the re, n the hrvesting time where the smples were ollete s well s the rying proess. Drying n the rying onitions (oxygen, high temperture, length of time, without vuum, et.) my use polymeriztion retions, reue some polyphenols n their ntioxint pity, n inue the formtion of new ompouns (Kenri et l., 2012). As stte y Kenri et l. (2012) tehin oxition mehnism initilly onstruts semiquinon, whih then onverts into quinon. The quinon ompoun n ret with mino i, protein, or other polymers to proue pronthoyniin. An then pronthoyniin n egre into tehin euse pronthoyniin represents n oligomer or polymer from flvn-3-ol (tehin/epitehin). These flvnols hve lrey een etete in some erry fruit n lef (Sellppn et l., 2002; Hrris et l., 2007; Vrhovsek et l., 2012; Deng et l., 2014). In lueerry skin n flesh, elphiniin, yniin, petuniin, peoniin, n mlviin monoglyosies re the min nthoynins. Beuse of the iversity in the glyosyltion n yltion pttern, more thn 25 nthoynins hve een ientifie in lueerries (Hrris et l., 2007; Moze et l., 2011). The mounts n istriution of nthoynins in the erries iffer epening on their plnt speies, ultivtion onitions in whih the fruit hve een grown or store (e.g. light, temperture), n prouing istrits, ue to geneti ifferenes mong wil n ultivte vrieties (Ehlenfelt n Prior, 2001). Blueerry leves re y-prouts of the lueerry inustry. The leves ontin in n exessively mount of polyphenols, whih retes n opportunity for their use in the neutreutil inustry, more so thn the fruits (Ehlenfelt n Prior, 2001; Kim n Um, 2011; Li et l., 2013; Deng et l., 2014). But no nthoynins hve een foun in fresh green leves (Hrris et l., 2007; Li et l., 2013). We foun fresh n rie V. myrtillus fruit extrts with high levels of mlviin-3-o-gluosie hlorie ( mg/kg FW n mg/kg DW) n yniin-3-o-gluosie hlorie ( mg/kg FW n mg/kg DW) ollete from Turkey, respetively. As mentione erlier, fresh n rie lueerry lef extrts hve signifintly less (p<0.05) nthoynins thn fruit extrts. Nevertheless, there were numerous reports tht nthoynins were etete in lueerry leves (Hrris et l., 2007; Kim n Um, 2011). These ifferenes re possily relte to the hrvesting seson of leves. Within the nthoyniin reutse or leuonthoyniin reutse tivity, the trnsriptionl ontrol fvors the expression of the nthoyniin synthse in sun-expose leves. The erese in pronthoyniin ontent in fvor of nthoynin proution hs lso een oserve uring fruit evelopment t the point when leves turne re, tivting the iosynthesis of yniin glyosies (Jkol et l., 2004; Li et l., 2013). In previous stuy reporte y Li et l. (2013) n Riihinen et l. (2008) nthoynins were not etete in green leves of lueerry n ilerry, ut low mount ws etete in re leves. Resvertrol is type of nturl phenol n phytolexin proue nturlly y severl plnts in response to injury or when the plnt is uner ttk y pthogens suh s teri or fungi (Frémont, 2000). The ourrene of resvertrol in Vinium erries shoul not e surprising, s its ourrene in the plnt kingom ppers to e wiespre (Rimno et l., 2004). Foo soures of resvertrol inlue the skin of grpes, lueerries, rsperries, n mulerries (Jsiński et l., 2013) n the vriility in the resvertrol ontent in fruit suh s lueerries hnges ue to foo proessing or preprtion. Reently, resvertrol ws reporte y Lyons et l. (2003) t levels of pproximtely ng/g smple (Rimno et l., 2004). The resvertrol ontents of lueerries n the relte ilerry, V. myrtillus L., were ultivte in severl ifferent geogrphil regions (Lyons et l., 2003). In our stuy, resvertrol ws foun in fresh fruit n lef extrts ( mg/kg FW n mg/kg FW, respetively) from oth regions. The previous t for trns-resvertrol ontent (0.4 mg/100 g FW) in lueerry (Moze et l., 2011) gree with ours, ut it hs lso een etermine to e lower for lueerries (Wng et l., 2008). Also, s stte y Lyons et l. (2003), lueerries n ilerries were foun to ontin resvertrol n the level of this hemoprotetive ompoun in these fruits ws <10% of tht reporte for grpes. In grpes, the levels of resvertrol were foun to vry with the time of hrvest, environmentl n limti onitions, mg GAE/kg Fresh V.myrtillus fruit extrts E1FRF E2FRF E3FRF K1FRF K2FRF K3FRF Totl Phenol mg GAE/kg Drie V.myrtillus fruit extrts e e E1DRF E2DRF E3DRF K1DRF K2DRF K3DRF Totl Phenol Fresh V.myrtillus lef extrts Drie V.myrtillus lef extrts mg GAE/kg e mg GAE/kg E1FRL E2FRL E3FRL K1FRL K2FRL K3FRL E1DRL E2DRL E3DRL K1DRL K2DRL K3DRL Totl Phenol Totl Phenol Fig. 1: Totl phenoli (TP) ontents (mg GAE/kg) of fresh n rie V. myrtillus fruit n lef extrts (E: Erek, K. Kpığ, FR: Fresh, DR: Drie, F: Fruit, L: Lef, 1-3: Coes of smples ollete from ifferent regions, Different letter(s) on r inite sttistilly signifint ifferenes, p<0.05)

7 Biotive ompouns in fruits n leves of lueerry 121 n plnt evelopmentl stge (Rimno et l., 2004). These ftors possiility ffete the ifferenes in resvertrol ontents of the V. myrtillus extrts in this stuy. Totl phenol n totl nthoynin ontents The TP ontents were signifintly ifferent mong the fruit n lef V. myrtillus extrts (Fig. 1). The TP ws mg GAE/kg in fresh fruit extrts, wheres in rie fruit were etermine to e mg GAE/kg. For the fresh n rie lueerry lef extrts, their TP ontents were signifintly (p<0.05) higher thn those in fresh n rie fruits. In the lef tissues of 87 highush lueerries, the men vlues of polyphenol ontents were ~30 times higher thn oserve in fruits on FW sis (Ehlenfelt n Prior, 2001). Skupień et l. (2006) reporte tht the TP ontents in V. orymosum L. lef extrts ws mg/100 g w rie leves. In ition, the TP of the lef extrts of V. myrtillus in Erek n Kpığ regions were signifitly higher (p<0.05) thn those of lkerry leves ( mg of GAE/g of DW) n strwerry leves (55.2 mg of GAE/g of DW) reporte y Wng n Lin (2000). Li et l. (2013) n Oszmiński et l. (2011) inite tht the polyphenol ontent of the lueerry leves ws muh higher thn those of ny other leves of teste erries (lkerry, rsperry, honeyerry, n strwerry). The ifferenes in TP ontents etween fruit n lef extrts were sttistilly signifint (p<0.05). Generlly, the TP of plnt extrts n e ffete y solvent, its polrity, its onentrtion, n/or extrtion metho metho (Deng et l., 2014; Xioyong n Luming, 2014). It is lso well known tht geneti, gronomi or environmentl ftors ply importnt roles in phenoli omposition n nutritionl qulity of rops (Yng et l., 2009). When the TP ontents of these extrts re ompre with white wines, these plnts oul ontriute the sme helth enefit s those wines in terms of polyphenols. A lrge vrition ws oserve mong fruit n lef extrts for TA ontent (Fig. 2), rnging from to mg/kg (in fresh fruit), from to mg/kg (in rie fruit), from to mg/kg (in fresh lef), n from to mg/kg (in rie lef). Fresh n rie fruit smples re goo soure of nthoynin (T. 1-4), however, fresh fruit smples reore ereses/inreses in TA ontent. The TA ontent in lueerry fruit n lef extrts in this stuy ws omprle to the quntity reporte y Ehlenfelt n Prior (2001), Sellppn et l. (2002), Lohhoompol et l. (2008), n Wng et l. (2015). Results otine from severl stuies suggest tht the TP n TA ontents in lueerry fruit re influene y the ultivr, hrvest time, the growing seson, fruit mss, mturity, environmentl growing onitions, growing lotion, posthrvest storge onitions, rying proess, ifferent extrtion methos, irrition, temperture, n pthogen ttks (Routry et l., 2014). Antioxint pity Blueerry fruit n lef show high ntioxint pity, orrelte espeilly with their nthoynin n other phenoli ompouns ontent, n my e onsiere s one of the highest ntioxint soures mong fruits n vegetles (Vrhovsek et l., 2012). The vlues foun in the fresh V. myrtillus fruit extrts for ntioxint pity (Fig. 3) were μmol TE/g FW y CUPRAC, μmol TE/g FW y DPPH, μmol TE/g FW y ABTS, n μmol TE/g FW y FRAP metho, vlues lower/higher thn or lose to those foun in fresh lef extrts (Fig. 4). Also in lueerry fruits n leves stuie y Wng n Lin (2000), Ehlenfelt n Prior (2001), Sellppn et l. (2002), Li et l. (2013), n Wng et l. (2015) using the sme methos, the V. myrtillus fruit n lef extrt results otine in this stuy were similr to those stuies. Therefore, s reporte y Wng n Lin (2000) n Prk et l. (2012), euse of their high ntioxint ontent, lueerry leves n lso e e to te mixes to inrese the ntioxint pity level in the everges for greter enefits to humn helth. Previous stuies foun iret reltionship etween the ntioxint pity, n the TP n TAs ontents in lueerry fruits n leves (Ehlenfelt n Prior, 2001; Li et l., 2013). In this stuy, there ws strong orreltion etween ntioxint pity n TP ontent Fresh V.myrtillus fruit extrts Drie V.myrtillus fruit extrts mg/kg mg/kg E1FRF E2FRF E3FRF K1FRF K2FRF K3FRF E1DRF E2DRF E3DRF K1DRF K2DRF K3DRF Totl Anthoynin Totl Anthoynin Fresh V.myrtillus lef extrts Drie V.myrtillus lef extrts mg/kg mg/kg E1FRL E2FRL E3FRL K1FRL K2FRL K3FRL 10 5 E1DRL E2DRL E3DRL K1DRL K2DRL K3DRL Totl Anthoynin Totl Anthoynin Fig. 2: Totl nthoynin (TA) ontents (mg/kg) of fresh n rie V. myrtillus fruit n lef extrts (E: Erek, K. Kpığ, FR: Fresh, DR: Drie, F: Fruit, L: Lef, 1-3: Coes of smples ollete from ifferent regions, Totl nthoynins were expresse s yniin-3-gluosie equivlents, Different letter(s) on r inite sttistilly signifint ifferenes, p<0.05)

8 122 N. Değirmenioğlu, O. Gürüz, G.E. Krtepe, R. Irkin Fresh V.myrtillus fruit extrts Drie V.myrtillus fruit extrts E1FRF E2FRF E3FRF K1FRF K2FRF K3FRF E1DRF E2DRF E3DRF K1DRF K2DRF K3DRF CUPRAC CUPRAC Fresh V.myrtillus fruit extrts Drie V.myrtillus fruit extrts E1FRF E2FRF E3FRF K1FRF K2FRF K3FRF E1DRF E2DRF E3DRF K1DRF K2DRF K3DRF DPPH DPPH Fresh V.myrtillus fruit extrts Drie V.myrtillus fruit extrts 2 18,00 16,00 14,00 12,00 1 8,00 6,00 4,00 2,00 E1FRF E2FRF E3FRF K1FRF K2FRF K3FRF 2 18,00 16,00 14,00 12,00 1 8,00 6,00 4,00 2,00 E1DRF E2DRF E3DRF K1DRF K2DRF K3DRF ABTS ABTS Fresh V.myrtillus fruit extrts Drie V.myrtillus fruit extrts 9,00 8,00 7,00 6,00 4,00 3,00 2,00 1,00 E1FRF E2FRF E3FRF K1FRF K2FRF K3FRF 9,00 8,00 7,00 6,00 4,00 3,00 2,00 1,00 E1DRF E2DRF E3DRF K1DRF K2DRF K3DRF FRAP FRAP Fig. 3: Antioxint pities (μmol TE/g) of fresh n rie V. myrtillus fruit extrts (E: Erek, K. Kpığ, FR: Fresh, DR: Drie, F: Fruit, 1-3: Coes of smples ollete from ifferent regions, Different letter(s) on r inite sttistilly signifint ifferenes, p<0.05) in the rie fruit n fresh lef extrts, while wek orreltion TA ontent in fresh fruit extrts (T. 5, is ville in Supplementry mteril). These results inite tht the phenolis, rther thn the nthoynins lone, ply n importnt role in ontriuting to the whole ntioxint pity. The reson my e tht nthoynins in fruit n leves oul trnsform into other types of phenolis, whih hve higher levels of ntioxint pity, vi the rying n smple preprtion proesses. As reporte y previously stuy, in the ry heting experiment, ll phenolis inluing nthoynins in the lueerry pome were ompletely egre to smll frgments tht h no ntioxint pity. Thus, the loss of the nthoynins n other phenolis strightforwrly linke to the erese of ntioxint pity of the ry-hete pome (Bener et l., 2013). Conlusion The rying metho ffets the qulity of the en prouts suh s olor, texture, rom, long with its hemil onstituents (Routry et l., 2014). Therefore, the ontrol metho hosen uring this stuy ws fresh storge fter vuum pkging. Oven-rie fruit n lef smples retine higher/lower mounts of phenoli ontent, TP n TA oring to the egree of resistne to the rying proess of the phenoli ompouns n the growing seson, suggesting tht enviromentl growing onitions, hrvesting ltitue, n length n type of rying time use for the rie smples when ompre to the mount otine from fresh smples. Also, nthoynins, flvonols, n pronthoyniins re lote minly in the peel while hyroxyinnmtes re foun in the flesh (Goling et l., 2001), therefore, it is elieve tht they re ffete y ifferent egrees

9 Biotive ompouns in fruits n leves of lueerry 123 Fresh V.myrtillus lef extrts Drie V.myrtillus lef extrts E1FRL E2FRL E3FRL K1FRL K2FRL K3FRL E1DRL E2DRL E3DRL K1DRL K2DRL K3DRL CUPRAC CUPRAC Fresh V.myrtillus lef extrts Drie V.myrtillus lef extrts E1FRL E2FRL E3FRL K1FRL K2FRL K3FRL E1DRL E2DRL E3DRL K1DRL K2DRL K3DRL DPPH DPPH Fresh V.myrtillus lef extrts Drie V.myrtillus lef extrts E1FRL E2FRL E3FRL K1FRL K2FRL K3FRL E1DRL E2DRL E3DRL K1DRL K2DRL K3DRL ABTS ABTS Fresh V.myrtillus lef extrts 14,00 12,00 1 8,00 6,00 4,00 Drie V.myrtillus lef extrts 14,00 12,00 1 8,00 6,00 4,00 2,00 2,00 E1FRL E2FRL E3FRL K1FRL K2FRL K3FRL E1DRL E2DRL E3DRL K1DRL K2DRL K3DRL FRAP FRAP Fig. 4: Antioxint pities (μmol TE/g) of fresh n rie V. myrtillus lef extrts (E: Erek, K. Kpığ, FR: Fresh, DR: Drie, L: Lef, 1-3: Coes of smples ollete from ifferent regions, Different letter(s) on r inite sttistilly signifint ifferenes, p<0.05) from n pplie rying metho, while the ntioxint pities of the smples inrese uner the sme onitions. If omine with the other protetive methos, oven-rying prove to e suitle metho for V. myrtillus smples preservtion euse the phenoli ompouns n their funtionl properties were either inrese or t lest erese. Consequently, V.myrtillus fruit n lef n e reommene s n ition to foo omposition, to inrese the ntioxint pity, euse of their high ntioxint properties. Aknowlegements The uthors re grteful to Blikesir University Sientifi Reserh Projets Unit (Projet No: ) for the finnil support of this reserh. Referenes Akoni, R.B., Kumr, P., Annpurn, A., Pujri, M., 2011: Protetive effet of rutin n nringin o sperme qulity in streptozotoin (STZ) inue type 1 ieti rts. Irn J. Phrm. Res. 10, oi: /jpp Apk, R., Gulu, K., Demirt, B., Ozyurek, M., Celik, S.E., Bektsoglu, B., Berker, K.I., Ozyurt, D., 2007: Comprtive evlution of vrious totl ntioxint pity ssys pplie to phenoli ompouns with the CUPRAC Assy. Moleules. 12, oi: / Bener, M., Shen, Y., Apk, R., Finley, J.W., Xu, Z., 2013: Relese n egrtion of nthoynins n phenolis from lueerry pome uring therml i hyrolysis n ry heting. J. Agri. Foo Chem. 61, oi: /jf Benzie, I.F.F., Strin, J.J., 1996: The ferri reuing ility of plsm

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