Determination of antioxidant activities of some apple cultivars

Transcription

1 Vol. 44, 2017 (3): Hort. Si. (Prgue) Determintion of ntioxint tivities of some pple ultivrs Lütfi Pirlk*, Gülüstn Ünüvr, Nil Ersoy Selçuk University, Fulty of Agriulture, Deprtment of Hortiulture, Selçuklu, Kony, Turkey *Corresponing uthor: Astrt Pirlk L., Ünüvr G., Ersoy N. (2017): Determintion of ntioxint tivities of some pple ultivrs. Hort. Si. (Prgue), 44: This stuy ws onute in orer to etermine the ntioxint tivities of ertin pple vrieties. Glxy Gl, Srlet Spur,, n Grnny Smith were hrterize in this stuy. The ntioxint tivities mong the vrieties were etermine vi three ifferent proeures. Antiril tivity figures of the pple vrieties were etween ( Glxy Gl ) n ( Srlet Spur ). Inhiition levels of helting tivity with Fe +2 vrie etween %; Srlet Spur n extrts h the highest inhiition with 55.50% n Grnny Smith extrt h the lowest with the level. Among the extrts showing H 2 lerne effet, Grnny Smith (62.54%) n Srlet Spur (44.67%) extrts h the highest inhiition levels. In this stuy onute on pple vrieties, totl phenoli mtter levels were etermine in Srlet Spur,,, Glxy Gl n Grnny Smith. Keywors: ntiril tivity; phenolis; DPPH Apple trees hve een grown in ll ontinents exept Antrti n re well suite to the temperte limte regions n tropil regions with high ltitues (Korn, Skirvin 1984). Turkey is epte s the geneti entre for other griulturl rops s well s fruit plnts n is rih in ifferent types of flor n ultivrs of fruit trees. Turkey is the originl growth ountry of most fruit plnts s well s pple tree. In reent yers, helthy foo onsumption hs gine gret importne for humn eings. Grpes, mong the fruits, ply very importnt role ue to high ontents of flvonois, phenoli is of whih some re nti-ner, or hve nti-mutgeni n ntioxint omponents. Antioxints eliminte the moleules lle free rils tht our in the humn oy fter the metoli tivities. In the se of exess free ril proution, they hve hrmful effet on ell, n ner my our from the tion of some enzymes. Antioxints re not proue only y oy ells, ut lso re tken from foo. The funmentl nturl ntioxints tht re present in foo tht protet the humn oy from the hrmful free rils re vitmins (C, E n A), flvonois, rotenois n polyphenols. There ws foun n inverse reltionship etween the onsumption of fruit n vegetle, n ner or hert ttk (Rie-Ivens et l. 1997). Therefore, it ws suggeste tht suh risk oul e reue y onsumption of foo with high ntioxint ontent (Snr 2004). Apple fruit is very importnt foo resoure for humns n most pple fruits re onsume fresh. In some ples, they re lso onsume s ry fruit, slie in ns, syrup, mrmles n jm. In reent yers, they hve een use for pst, fruit juie n vinegr setors. Apple fruit hs minerl slts n vitmins so it is very importnt soure of foo for humn. It my e helpful in reution of risks of some iseses suh s some ners, hert prolems, sthm n ietes ue to its ontent of rih hemil ompouns. Lortory tests inite tht pple fruit onsumption inhiite expnsion of ner ells n reue the lipi oxi- 120

2 Hort. Si. (Prgue) Vol. 44, 2017 (3): tion n holesterols (Leontowiz et l. 2002). After lueerry, it hs the highest ntioxint tivities (Boyer, Liu 2004). A numer of epiemiologi stuies showe tht there is negtive reltionship etween hert-vessel or ner iseses n fruit-vegetle onsumption (Blok et l. 1992; Liu 2003). In reent yers, onsumption of foo with high ntioxint pity hs een suggeste (Hlliwell 2001); it is known tht fruits, very importnt for helthy life, hve gret mounts of ntioxints (Vinson et l. 2005). There re ifferent mounts n types of phenoli ompouns (seonry metolites), whih protet the rops from some hrmful mterils. Phenoli ompouns foun in foo re lssifie into two group nmely phenoli is (phenoli roni is) n flvonois (flvone erivtives). Phenoli ompouns re very importnt sine they re sustrte resulting in lk olouring in fruit n vegetles (Eskin et l. 1976), they ret with metl ions using olour hnges (Herrmnn 1976), they use unesire tstes in foo (Le 1984), n they ret with proteins leing to preipittion (Oh, Hoff 1987). Apple is the fruit with the seon highest phenolis ontent fter lueerry. In preventing lung ner, it is rnke thir fter lueerry n lemon. Apple lso rnke the seon in totl onentrtion of phenoli ompouns, n perhps more importntly, pples h the highest portion of free phenolis when ompre to other fruits (Sun et l. 2002). To solve the helth prolems espeilly in eveloping ountries, plnt prouts were use wiespre ue to the lrge mount of uget requirement of phrmologil prouts. There re mny plnts use for ifferent purposes y the puli in Turkey ut their sientifi tivities re still unknown. There re some stuies showing ntioxint effetiveness of plnt extrts n plnt prouts (Çon et l. 2003; Coulis et l. 2003; Tepe et l. 2006; Altun et l. 2007; Krtl et l. 2007). For this reson, pple ultivrs foun in Turkey were stuie. MATERIAL AND METHODS Glxy Gl, Grnny Smith,, Srlet Spur n pple ultivrs grfte onto M9 rootstok were use. The fruit smples were pike t the iel ommeril mturity from the Glxy Gl, Grnny Smith,, Srlet Spur n pple ultivrs t the time of hrvest. The mturity is se on the etermintion of firmness, refrtometri vlue n strh onversion. Antioxint nlysis ws performe t the lortory of Deprtment of Foo Engineering, Afyon Kotepe University. The stuy esign omprise three replites with ten fruits for eh nlysis. For investigtions, pple fruits were thinly setione from skin to the fruit entre. Determintion of ntioxint tivities Smple preprtion. 2.5 g fruit tissue from eh rie plnt smple were extrte y homogenizing in mixer (Ultr Turrx; IKA-Werke GmH & Co., Germny) tue isperser (IKA, Stufen, Germny) with 50 ml solvent (50% wter-methnol). Extrts were filtere thorough pper (Filter- L; Inoi Filters, S.A., Spin), then entrifuge t 4,000 g 3 min t 4 C. 50 ml of superntnt ws further filtere using lue n, No. 589 filter pper (Shleiher n Shuell, Green rion; Shleiher n Shuell & GmH, Germny). Determintion of free ril svenging tivity. To mesure the free ril svenging tivity of 1.1-iphenyl-1-pirylhyrzyl (DPPH) moifie ssy of Brn-Willims et l. (1995) ws use. For this purpose, ilutions in the rnge of etween 0.4 n 4 mg/ml were prepre from the juie extrtions. The ilutions etween 0.4 to 4 mg/ml were prepre with methnol. 3.9 ml DPPH solution prepre with M (molr) methnol ws e to eh 0.1 ml of ilution n ws shken well. Those smples were kept 60 min in the rk room uner room temperture. Asorne ws then mesure ginst methnol stnr t 515 nm. Smples without DPPH were use s ontrol tretment. All nlyses were rrie out in uplite. The tivity ws lulte utilizing stnr urve. The liner regression eqution ws erive y mesuring the sorption in 515 nm from seven ifferent onentrtions of DPPH ( M): A(515 nm) = (C DPPH) (R 2 = 0.961) The resiul DPPH onentrtions s % were lulte s: % Resiul DPPH = (DPPH) smple /(DPPH) The regression eqution ws evelope y iviing resiul DPPH * to DPPH * of smple uner test 121

3 Vol. 44, 2017 (3): Hort. Si. (Prgue) mei. By use of tht eqution, smple onentrtion reuing 50% of initil DPPH * onentrtion, (effiient onentrtion = EC 50 ), ws otine for rop smples. By iviing EC 50, 1/EC 50, ntiril tivity (AE) ws lulte. Clultion of Fe +2 helting tivity. Chelting tivity of pple smples were mesure y methos moifie y RIVAL et l. (2001). 1 ml of extrt with ifferent onentrtions etween 6 45 mg/ml n 3.7 ml eionize wter were mixe. A 0.1 ml 2mM FeCl 2 solution ws e, shken n kept in the rk t room tempertures for 70 min. After tht, 0.2 ml 5mM ferrozin ws e n mixe gin. The resulting Fe +2 ferrozin omplex sorne t 562 nm were mesure fter 10 minutes. In the ontrol, 1 ml wter ws use inste of smple. The helting pity of smples ws lulte s in Yen n Wu (1999): % helting pity = [1 (smple sorne/ /ontrol sorne)] 100 Mesurement of H 2 svenging. H 2 svenging pility of rops n hers n e ignose spetrometrilly (Ruh et l. 1989). For tht, 1 ml (2, 6 n 10 mg/ml) smple, 3.4 ml 0.1M phosphte uffer solution (ph 7.4) n 0.6 ml 43mM H 2 were mixe n fter 60 min the sorne of the mixture ws mesure t 230 nm. Control smples ontining no H 2 solution were prepre for eh smple. A liner regression eqution ws evelope to etermine the H 2 (mm) onentrtion. For tht, 0.6 ml 10, 15, 25, 43 n 50mM H 2 were e to the 3.4 phosphte uffer solution n their sorne ws mesure t 230 nm. The following liner regression eqution ws evelope: A(230) = C (H 2, mm) (R 2 = 0.961) Removing H 2 pity of smples ws lulte s: H 2 removing pity (%) = [1 ( H 2 of smple/ H 2 of ontrol)] 100 Clultion of totl phenoli ontent. The totl phenoli ontent of smples ws nlyse y using the Folin Ciolteu olorimetri metho (Singleton, Rossi 1965). Reings were performe spetrophotometrilly t 765 nm. Vlues were lulte s mg GA/g. Totl phenoli mtter ontent ws presente s glli i (GA) equivlent. Sttistil nlysis. The results were evlute using the R- progrm with the Dunn test t 5% signifine level in orne with the Rnomize Prel Tril esign (Anonymous 2013). RESULTS AND DISCUSSION The ntioxint pity of five pple ultivrs grown in Turkey ws mesure y four ifferent ssys to etermine the reltive level of potentilly helth-enefiil spets of eh of the ultivrs. Eh ssy mesure ifferent spet of pity from eing le to reue free rils to totl phenoli ontent. Determintion of ntioxint tivity Free ril svenging pity. Antioxint pity ws mesure y the mount of smple ple to reue the DPPH onentrtion to 50% of the initil mount with lower vlues eing n initor of high ntioxint pity. From tht vlue the ntiril effiieny or ntiril tivity ws lulte (Molyneux 2004). The present stuy showe tht ntiril tivity, AE, ws in inresing mounts: 0.545% for Grnny Smith, 0.592% for Glxy Gl, 0.675% for, 0.737% for n 0.802% for Srlet Spur (Fig. 1). Chelting tivity of Fe +2 Chelting tivity for metl n e evlute y the ompetitions etween plnt extrts n ferrozin for ining Fe +2 ions. The inhiition grph showing the metl helte potentil s perentge of plnt extrt otine from ll the pple ultivrs is shown in Fig. 2. The highest vlues were otine from Srlet Spur t 55.20% n t 53.18% while the lowest vlue ws lulte s 40.20% from Grnny Smith. H 2 svenging The oxition of H 2 y the plnt extrts ws etermine y using the metho s suggeste y Ruh et l. (1989). Although none of the extrts 122

4 Hort. Si. (Prgue) Vol. 44, 2017 (3): Srlet Spur Grny Smith Glxy Gl Fig.1. Antiril tivity for ifferent pple ultivrs (%) (LSD: ) ompletely oxiize the H 2, they i rnge from 44.67% to 62.53% (Fig. 3). Phenoli ontent e Phenoli ontents s the equivlent of glli i in reserh pples were lulte. As seen in Fig. 4, the highest phenoli ontents were foun in Srlet Spur t 26.86% n, gin, Grnny Smith h the lowest level t 18.29%. The phenoli n flvonoi ontents highly epen on pple ultivrs. Therefore, ifferent pple ultivrs hve ifferent ntioxint tivity (Boyer, Liu 2004). Apples, like other fruits, vry in hemil omposition even within the sme vriety, epening on mturity, lotion proue, n griulturl prties, s well s numerous other environmentl ftors (Lee et l. 2003). Inee, signifint vritions in phenoli ontent n ntioxint tivity were oserve mong ultivrs n even mong ifferent fruits in the sme ultivr (Imeh, Khokhr 2002). Flvonoi ontents of Golen Deliious, Reinet, Re Deliious n Grnny Smith pple ultivrs were ompre n Golen Deliious h the Srlet Spur Grnny Smith Glxy Gl Fig. 3. H 2 svenging (%) (LSD: ) lowest, n Reinet pple ultivr h the highest flvonoi ontent followe y Grnny Smith n Re Deliious pple ultivrs (Esrp n Gonzles, 1998). Drogoui et l. (2008) mesure the phenoli n sori i ontents of, Golen Deliious, Grnny Smith, Jongore, Mutsu, Strkrimson n Fyriki (lol ultivr of Greee) in pple skin. The highest ntioxint pity of 35.7 mg/g n phenoli mtter ontent of 19.9 mg/g were otine from Strkrimson n ws followe y Jongore, Mutsu, Fyriki n. The skin of Golen Deliious (13.8 mg/g) n Grnny Smith (8.4 mg/g) h the lowest ntioxint pity. In the nlysis of fruit flesh, the highest ntioxint pity ws otine from Fyriki s 11.9 mg/g or 9.8 mg/g n tht ws followe y Jongore, Mutsu n Strkrimson 5.6 mg/g n 5.4 mg/g. The lowest ntioxint pity in fruit flesh ws foun in, Golen Deliious n Grnny Smith (etween 3.7 mg/g n 3.5 mg/g). The other importnt results re tht the ntioxint pity of fruit skin ws three fol higher thn the fruit flesh n the highest sori i ontent mong the pple ultivrs ws otine from Fyriki (4.4 mg/g). Srlet Spur Grnny Smith Glxy Gl Fig. 2. Chelting tivity of Fe +2 (%) (LSD: ) Srlet Spur Grnny Smith Glxy Gl Fig. 4. Phenoli ontent (mg GA/g) (LSD:

5 Vol. 44, 2017 (3): Hort. Si. (Prgue) In the present stuy, the ntioxint pity ws foun in eresing orer in Srlet Spur, Pink Ly,, Glxy Gl n Grnny Smith pple ultivrs. All four mesurements were in greement with this orer. Apple is rih in nthoynins n phenoli mtter ontent mong the fruits n vegetles. Thus, pple fruit shoul e e to the iet menu to protet the humn oy from the oxitive stress. In ition, pple plnts re grown very wiely in Turkey n it is thought tht pple fruits hve some other possile useful effets for our helth. Tht issue shoul e reserhe in etils in orer to minimize the puli helth uget. Referenes Altun M.L., Çıtoğlu G.S., Yılmz B.S., Çon T. (2007): Antioxint properties of Viurnum opulus n Viurnum lntn growing in Turkey. Interntionl Journl of Foo Siene n Nutrition, 5: 1 6. Anonymous (2013): The R Projet for Sttistil Computing. Aville t Blok G., Ptterson B., Sur A. (1992): Fruit, vegetles, n ner prevention: review of the epiemiologil eviene. Nutrition n Cner, 18:1 29. Boyer J., Liu R.H. (2004): Apple phytohemils n their helth enefits. Nutrition Journl, 3: 5. Brn-Willims W., Cuvelier M.E., Berset C. (1995): Use of freeril metho to evlute ntioxint tivity. Leensmittel Wissenshft un Tehnologie, 28: Coulis M., Tzkou O., Verykokiou E., Hrvl C. (2003): Sreening of some Greek romti plnts for ntioxint tivity. Phytotherpy Reserh, 17: Çon T., Sltn Çıtoğlu G., Sever B., Sn M. (2003): Antioxint tivities of plnts use in tritionl meiine in Turkey. Phrmeutil Biology, 41: D Silv Porto P., Lrnjinh J., De Freits V. (2003): Antioxint protetion of low ensity lipoprotein y proyniins: struture/ tivity reltionships. Biohemil Phrmology, 66: Drogoui D.P., Mihiliis Z., Pneliis G. (2008): Peel n flesh ntioxint ontent n hrvest qulity hrteristis of seven pple ultivrs. Sienti Hortiulture, 115: Esrp A., Gonzlez M.C. (1998): High-performne liqui hromtogrphy with ioe-rry etetion for the etermintion of phenoli ompouns in peel n pulp from ifferent pple vrieties. Journl of Chromtogrhy A, 823: Eskin N.A.M., Henerson H.M., Townsen R.J. (1976): Biohemie er Leensmittel. Hüthig Verlg. Griffits H.R., Lune J., Aroum O.I., Hlliwell B. (1998): Moleulr Biology of Free Rils in Humn Diseses. Lonon, Oi Interntionl. Hlliwell B. (2001): Foo-erive ntioxints: How to evlute their importne in foo n in vivo. Hnook of Antioxints. Los Angeles, New York, CRC Press. Imeh U., Khokhr S. (2002): Distriution of onjugte n free phenols in fruits: ntioxint tivity n ultivr vritions. Journl of Agriulturl n Foo Chemistry, 50: Krmn Ş. (2008): Türkiye e yetiştirilen zı elm çeşitlerinin toplm ntioksin kpsitelerinin ve ntioksin özellik gösteren şlı ileşenlerinin krşılştırılmsı. [MS Thesis.] İstnul Üniversitesi Fen Bilimleri Enstitüsü Kimy Anilim Dlı, İstnul, Korn S.S., Skirvin R.M. (1984): Nomenlture of the Cultivte Apple. HortSiene, 19: Lee K.W., Kim Y.J., Kim D., Lee H.J., Lee C.Y. (2003): Mjor phenolis in pple n their ontriution to the totl ntioxint pity. Journl of Agriulturl n Foo Chemistry, 51: Leontowiz H., Gorinstein S., Lojek A., Leontowiz M., Ciz M., Soliv-Fortuny R., Prk Y., Jung S., Trkhtenerg S., Mrtin-Belloso O. (2002): Comprtive ontent of some iotive ompouns in pples, pehes, n pers n their influene on lipis n ntioxint pity of rts. The Journl of Nutritionl Biohemistry, 13: Liu R.H. (2003): Helth enefits of fruit n vegetles re from itive n synergisti omintions of phytohemils. Amerin Journl of Clinil Nutrition, 78(3 Suppl): 517S 520S. Miller N.J., Rie-Evns C.A. (1996): The reltive ontriutions of sori i n phenoli ntioxints to the ntioxint tivity of ornge n pple frui juies n lkurrnt rink. Foo Chemistry, 60: Molyneux P. (2004): The use of the stle free ril iphenylpiryl-hyrzl (DPPH) for estimting ntioxint tivity. Songklnkrn Journl of Siene n Tehnology, 26: Oh H., Hoff J.E. (1987): ph epenene of omplex formtion etween onense tnnins n proteins. Journl of Foo Siene, 52: Rie-Ivens C.A., Miller N.J., Pgng G. (1997): Antioxint properties of phenoli ompouns. Trens in Plnt Siene, 2: Rivl S.G., Boeriu C.G., Wihers H.J. (2001): Cseins n sein hyrolystes ntioxitive properties n relevne to lipoxygense inhiition. Journl of Agriulturl n Foo Chemistry, 49: Ruh R.J., Cheng S.J., Kluning J.E. (1989): Prevention of ytotoxiity n inhiition of intrellulr ommunition 124

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