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Cinc. ecnol. Aliment., Campinas, 28(3): 702-708, jul.-set. 2008702

ISSN 0101-2061 Cincia e ecnologia de Alimentos

Recebido para publicao em 6/9/2007Aceito para publicao em 22/1/2008 (002827)1 Departamento de Alimentos e Nutrio Experimental, Universidade de So Paulo USP, Av. Pro. Lineu Prestes, 580, Bloco 14, CEP 05508-900, So Paulo - SP, Brasil,

E-mail: [email protected] Empresa de Pesquisa Agropecuria de Minas Gerais EPAMIG, Ncleo Tecnolgico EPAMIG Uva e Vinho, Fazenda Experimental Caldas FECD, Av. Santa Cruz, 500,

CP 33, CEP 37780-000, Caldas - MG, Brasil*A quem a correspondncia deve ser enviada

Physico-chemical characterization and bioactive compounds

of blackberry fruits (Rubus sp.) grown in Brazil

Caracterizao sico-qumica e de compostos bioativos em amora-preta (Rubus sp.) cultivada no Brasil

Neuza Mariko Aymoto HASSIMOO1, Renata Vieira da MOA2,Beatriz Rosana CORDENUNSI1*, Franco Maria LAJOLO1

1 IntroductionFruit and vegetable intake has proven to reduce the develop-

ment o a considerable number o chronic diseases, such as can-cer and cardiovascular diseases. Te protective eect evidencedin laboratory and epidemiological studies has been associatedwith a variety o nutrient and non-nutrient constituents, beingmany o them characterized by their antioxidant properties(PIEA, 2000). Among the compounds with high antioxi-dant capacity are ascorbic acid, tocopherols, -carotene andphenolic compounds, especially avonoids (DREWNOWSKI;GOMEZ-CARNERO, 2000).

Anthocyanins, the glycoside orm o the anthocyanidins,

belong to a class o avonoids which present water-soluble pig-ments responsible or the orange, red and blue colors o manyruits, vegetables, leaves, owers and roots. Anthocyanidinsvary with dierent hydroxyl or methoxyl substitutions in theirbasic structure, avylium (2-phenylbenzopyrillium). Only sixo the 16 anthocyanidins identied in natural products occur

requently and in many dierent vegetables and ruits: cyanidin,pelargonidin, delphinidin, peonidin, petunidin and malvidin(WU; PRIOR, 2005a; b). Anthocyanins are highly studied com-pounds because o their eects on human health as antioxidantcompounds, as shown in a number o in vitro and in vivo studies(KHKNEN; HEINONEN, 2003), or their anti-inammatoryeect modulating ciclooxygenase 1 and 2 enzymes (ALL et al.,2004) and their inhibitory eect on the development o somecancer cells (HAGIWARA; YOSHINO; ICHIBARA, 2002).

Among the small ruits group, including strawberry, rasp-berry, blueberry and blackberry crops, blackberry appears as

the richest source o anthocyanins, with cyanidin content rang-ing rom 93 to 280 mg.100 g1 resh weight depending on thecultivar (CONNOR et al., 2002), ollowed by black raspberry(up to 197 mg), blackberry (153 mg) and strawberry (32 mg)(WANG; LIN, 2000). Blackberry plants were introduced inBrazil by the Estao Experimental de Pelotas (RS), renamed

Resumo

Cinco cultivares de amora-preta (Rubus sp.) oram avaliadas quanto a sua capacidade antioxidante, perl de compostos bioativos e composiosico-qumica. Os nveis de cido ascrbico total, presentes na orma de cido desidroascrbico, variaram entre 9,8 a 21,4 mg.100 g1 (b.u.).Os principais avonides presentes nas cinco cultivares oram: a antocianina cianidina (66 a 80% do total de avonides); o avan-3-olepicatequina; e os avonis quercetina e traos de caenerol. As cinco cultivares apresentaram alta capacidade antioxidante quando avaliadaspelo sistema de co-oxidao -caroteno/cido linolico, similar ao antioxidante sinttico BH, na concentrao de 50 M. A cultivar Guaraniapresentou os maiores teores de avonides totais, antocianina total, cianidina e de capacidade antioxidante, enquanto que a cultivar Caigangueapresentou alto contedo de vitamina C e de enlicos totais. Estas duas cultivares tambm apresentaram uma boa correlao SS/A. Assim,a amora-preta pode ser considerada uma boa onte de compostos bioativos e assim contribuir com a manuteno do estado de sade.Palavras-chave: Rubus; avonides; antocianinas; capacidade antioxidante.

Abstract

Five blackberry cultivars (Rubus sp.) were evaluated or antioxidant capacity, bioactive compounds and composition. Ascorbic acid levels,consisting o dehydro-ascorbic acid, ranged rom 9.8 to 21.4 mg.100 g1 resh weight. Cyanidin (66 to 80% o total avonoids), epicatechin,quercetin and traces o kaemperol were the main avonoids ound in all cultivars. Te ve cultivars presented high antioxidant capacity inthe -carotene/linoleic acid system, with inhibition similar to the synthetic antioxidant BH, at a 50 M concentration. Caingangue cultivarpresented high vitamin C and total phenolics content, while Guarani had the highest cyanidin, total anthocyanin and total avonoids levelsand also the highest antioxidant capacity. Tese cultivars also presented good SS/A ratios. From the data, at a quantitative level, blackberrycan be considered a good source o bioactive compounds, as well as potentially benecial to human health.Keywords:Rubus; avonoids; anthocyanin; antioxidant capacity.

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Hassimotto et al.

EMBRAPA Clima emperado in the 1970s, and since thenits production has increased in the states o Rio Grande doSul, So Paulo and Minas Gerais, with the introduction andadaptation o new cultivars. Blackberry (Rubus eubatus) has ashrub rame with erect or trailing stems. Te blackberry ruits,weighing around 4 to 7 g, have an acid to sweet-acid taste andare not berries, but an aggregate ruit composed o many smaller

ruits called drupes. Blackberry, being easy to handle and havinglow production costs even concerning chemicals, is an alterna-tive crop or amily agriculture. Te resh ruits, besides beingvery nutritious, rich in minerals, pro-vitamin A, vitamin B andcalcium, also contain bioactive compounds, such as ellagic acidand anthocyanins (ANUNES et al., 2003).

Te antioxidant capacity o the dietary intake o ruitsand vegetables changes considerably according to the pres-ence o diverse compounds with antioxidant capacity, but theamount o anthocyanins present in them seems to be important(HASSIMOO; GENOVESE; LAJOLO, 2005). Among theBrazilian vegetables already studied, mulberry (Morus nigra),aa (Euterpe oleracea), jambolo (Syzygium cumini), purplelettuce (Lactuca sativa), purple cabbage (Brassica oleracea)and commercial rozen blackberry (Rubus sp.) pulp appear asimportant anthocyanin sources (HASSIMOO; GENOVESE;LAJOLO, 2005).

Species o the Rubus genus are known as natural sourceso anthocyanins with high antioxidant eect (DEIGHONet al., 2000). Due to the increasing interest in blackberry con-sumption and the knowledge that the nutritional compositionvaries according to the cultivar, this work aimed at evaluatingthe phenolic and anthocyanin content, antioxidant capacityand physicochemical quality o ve blackberry cultivars grownin Brazil.

2 Materials and methods

Blackberry ruits (Rubus eubatus) rom the cultivars upy,Guarani, Caingangue, Brazos and Seleo 97 were harvestedat the germplasm bank o Estao Experimental de Caldas(EPAMIG/FECD), Minas Gerais, Brazil. Te ripe ruits (blackcolor) were harvested during the 2003/2004 harvest season.Aer harvest, the ruits were selected, separating those withinjuries. Tey were then washed in chlorinated water, drainedand rozen, and stored at 20 C until analyses. Each sampleconsisted o ruits harvested at the beginning, middle and endo the season. Frozen ruits rom each sample were macerated

with liquid nitrogen using a mortar and pestle and immediatelysubmitted to analyses o ascorbic acid, avonoids, total phenolicsand the in vitro antioxidant assay.

Physicochemical characteristics: Frozen ruit samples werethawed at room temperature and homogenized in a mixer.Following seeds removal, 10 g o pulp rom each sample werepoured into a 150 mL vial and 80 mL o distilled water wasadded. Te vials were placed in a water bath at 80 C or 2 hours.Te extract was transerred to 100 mL volumetric asks andltered through a cheese cloth. itratable acidity was deter-mined by titration with 0.1 N NaOH to a pink color using 1%phenolphthalein as indicator and expressed as g.100 g1 citric

acid. Te pH o the diluted pulp o each sample was determinedusing a pH-meter and soluble solids (Brix) were determinedusing a handheld reractometer. Moisture and ashes were de-termined in the undiluted pulp, according to AOAC methods(1995). otal carbohydrates were determined in the diluted pulpaccording to Dubois et al. (1956). Soluble sugars were extractedthree times with 80% ethanol at 80 C. Aer centriugation, the

supernatants were combined and the ethanol was evaporatedunder vacuum. Te soluble sugar content was analyzed by highperormance liquid chromatography with pulse amperometricdetection (HPLC-PAD Dionex, Sunnyvale, Caliornia, USA),using a PA

1column (Dionex, Sunnyvale, Caliornia, USA) in an

isocratic run o 18 mM NaOH during 25 minutes. otal solublesugars were taken as the sum o glucose, ructose and sucrosevalues. All samples were analyzed in triplicate and the resultswere subjected to statistical analysis o variance. reatmentmeans were separated using the ukey test (p < 0.05).

otal anthocyanins: An aliquot o 2 g o undiluted pulpwas extracted with 20 mL acidied methanol (1% HCl, v/v),in a shaker, or 30 minutes, at room temperature, and storedovernight at 4 C. Te extract was ltered and washed severaltimes until complete removal o pigments and the volume wasadjusted to 100 mL in volumetric asks. Aer appropriate dilu-tion with acidied methanol, samples were stored in the darkor 2 hours and the absorbance was determined at 532 nm ina spectrophotometer. Anthocyanin concentration (expressedas mg pigment.g1 pulp) was determined using the molecu-lar weight (449.2) and molar absorbance (26,900) values orcyanidin-3-glucoside (LIMA et al., 2003; PERKINS-VEAZIE;COLLINS, 2002).

Ascorbic Acid (AA) content was determined according tothe method o Rizzolo et al. (1984). AA was extracted with meta-

phosphoric acid (0.3% w/v) and analyzed by reversed-phaseHPLC, in a Hewlett-Packard 1100 system, with an auto-samplerand a quaternary pump, coupled to a diode array detector.Te column used was a -Bondapack (300 mm x 3.9 mm i.d.,Waters, Milord, MA, USA) column, and elution (ow rate o1.5 mL/minute) was perormed in isocratic conditions with0.2 M sodium acetate/acetic acid buer (pH 4.2), monitoredat 262 nm. otal AA was estimated aer reduction o dehydro-ascorbic acid (DHA) with 10 mM dithiothreitol.

Flavonoid identication and quantication was perormedin duplicate according to the method o Hassimotto, Genoveseand Lajolo (2007), with slight modiications. he sample

was extracted three times in 100 mL methanol/water/aceticacid (70:30:5 v/v), or 2 minutes (Brinkmann homogenizer,Polytron-Kinematica GmbH, Kriens-Luzern, Sweden), in anice bath. Te homogenate was ltered under reduced pressurethrough lter paper (Whatman n 06). Te extract was concen-trated until methanol elimination, under vacuum, at 40 C, ina rotary evaporator (Rotavapor RE 120, Bchi, Flawil, Sweden)and made up with distilled water or application in solid-phaseextraction (SPE) columns. An aliquot o the extract was passedthrough polyamide (CC 6, Macherey-Nagel, Germany) col-umns (1 g/6 mL), previously conditioned with methanol anddistilled water. Impurities were washed out with distilled waterand retained avonoids were eluted with methanol containing

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Blackberry ruit composition

0.1% HCl. Te ow rate through the columns was controlledby means o a vacuum maniold Visiprep 24 DL (Supelco,Belleonte, PA, USA). Te eluate was evaporated to drynessunder reduced pressure at 40 C, redissolved in methanol: aceticacid (99:5 v/v), and ltered through a 0.22 m tetrauoroethyl-ene (PFE) lter (Millipore Ltd., Bedord, MA, USA), prior toHPLC analysis. Identication and quantication o avonoids

were achieved using analytical reversed-phase HPLC, in a Hewl-ett-Packard 1100 system, with an auto-sampler and quaternarypump, coupled to a diode array detector (DAD). Te columnused was a Prodigy 5 ODS-3 reversed-phase silica (250 mm4.6 mm i.d., Phenomenex Ltd.) column. Te solvents were (A)water/tetrahydrouran/triuoroacetic acid (98:2:0.1 v/v) and(B) acetonitrile, and the solvent gradient consisted o 8% Bat the beginning, 10% at 5 minutes, 17% at 10 minutes, 25%at 15 minutes, 50% at 25 minutes, 90% at 30 minutes, 50%at 32 minutes, 8% at 35 minutes (run time, 35 minutes). Elu-ates were monitored at 270, 370 and 525 nm. Flow rate was1 mL/minute, column temperature was 30 C, and injectionvolume was 5-20 L. Samples were injected in triplicate, and

lavonoids were quantiied using external standards. Peakidentication was perormed by comparison o retention timesand spectra with the standards and the library spectra. Resultswere expressed as milligrams aglycone per 100 g Fresh Weight(FW) (quercetin and cyanidin, Extrasyntese, Genay, France;epicatechin, Sigma, St. Louis, MO, USA), as mean standarddeviation (SD).

otal phenolics were measured in duplicate sampleso each extract according to the method o Zielinski andKozolwska (2000), using the Folin-Ciocalteu reagent and gal-lic acid as standard. Te results were expressed as Gallic AcidEquivalents (GAE).

In vitro antioxidant assay: Te antioxidant capacity othe extracts obtained rom the analysis o total phenolics wasdetermined according to the -carotene bleaching method(HASSIMOO; GENOVESE; LAJOLO, 2005). An aliquot(20 L) o-carotene solution (2 mg.mL1 in chloroorm) wasadded to a ask containing 40 L linoleic acid, 1.0 mL chloro-orm and 0.4 mL ween 40, and mixed. Te chloroorm wasevaporated to dryness under nitrogen. Aer this, oxygenateddistilled water (100 mL) was added and the mixture was shaken.Aliquots (100 L) o the extract were added to 2.9 mL o the-carotene solution in a cuvette and mixed. Te absorbance othe solution at 470 nm was measured immediately and aer2 hours o incubation in a water bath at 50 C using a Hewllet

Packard 8453 spectrophotometer. Te control consisted o100 L methanol instead o the sample extract. Antioxidantcapacity was calculated as percent inhibition relative to thecontrol.

3 Results and discussion

Physicochemical characteristics o the ive blackberrycultivars analyzed are described in able 1. Moisture contentwas around 90%, with dierences among cultivars being lowerthan 3%. Ash was close to 0.3% o total resh ruit weight inall cases. In terms o total soluble solids (Brix), Guarani andSeleo 97 cultivars had the highest values, while Brazos and

upy cultivars presented the lowest soluble solids contents,which resulted in low SS/A ratios (4.0 and 5.2, respectively),when compared to the mean SS/A ratio o 5.8 obtained orthe other cultivars (able 1). Te best SS/A ratio (7.4) wasobserved or Seleo 97 cultivar.

Soluble sugars ranged rom 2.7 to 4.8%, and were composedmainly o ructose and glucose (able 2), except or Seleo 97

cultivar that showed small amounts o sucrose (0.7 g.100 g1resh ruit). Glucose and ructose concentrations varied amongcultivars, but the glucose/ructose ratio was between 1.2 and 1.4or all cultivars. Te values or glucose and ructose were similarto other so ruits (SEYMOUR, 1996).

Vitamin C, total phenolics and total anthocyanins: Tetwo orms o ascorbic acid with vitamin C biological activi-ties were analyzed, the reduced orm (L-ascorbic acid AA)and the oxidized orm (L-dehydro-ascorbic acid DHA). Ingeneral, DHA content in vegetables represents less than 10%o total vitamin C (LEE; KADER, 2000), but in the ve black-berry cultivars studied, the reduced orm was not detected, only

the oxidized orm. otal ascorbic acid content (AA) rangedrom 9.86 mg.100 g1 FW (cv. Brazos) to 21.36 mg.100 g1 FW(cv. Caingangue) (able 3), which represent amounts highenough to consider blackberry as a good source o vitamin C.Te absence o AA in all cultivars under study may be relatedto the ast oxidation o vitamin C and absence o de novosynthesis o ascorbic acid during development or ripening.otal phenolics ranged rom 341 to 499 mg.100 g1 FW andCaingangue cultivar stood out with the highest values. otalanthocyanins determined by the colorimetric method rangedrom 116 to 194 mg.100 g1 FW (able 3).

Flavonoids and antioxidant capacity: Flavonoid content inblackberry cultivars ranged rom 123 to 213 mg.100 g1 resh

ruit (able 4). Anthocyanins were the main avonoids in black-berry ruits composition. Cyanidin derivatives (3 or 4 peaks)were the pigments detected in the ve cultivars, representing66-80% o the avonoids. Using a standard, the main cyanidinderivative peak was identied as cyanidin-3-O-glucoside. Tehighest cyanidin concentrations were detected in Guaraniand Caingangue cultivars (about 150 mg.100 g1), valueshigher than those detected or domestic and wild speciesgrown in China (1-118 mg.100 g1) (DEIGHON et al.,2000). Also, data shown or blackberry ruits are superior toanthocyanin contents ound in other red ruits, such as straw-berry (30 to 60 mg.100 g1, CORDENUNSI et al., 2003), purpleSurinam cherry (16.23 mg.100 g1, LIMA et al., 2005) or acerola(3.79 to 59.74 mg.100 g1, LIMA et al., 2003).

otal anthocyanin content determined by spectrophotometry(able 3) was higher when compared to cyanidin content deter-mined by HPLC. In general, the dierent color shades deter-mined by anthocyanins depend on the aglycone type, glycosila-tion, pH and co-pigmentation with organic anions (SAFFORD,1990). Te combination o these actors contributes to theobserved inal color, which does not represent the HPLCquantication o anthocyanin. Tus, color and color intensityo each colored vegetable are not necessarily associated with adistinct chromophore (aglycone), but with a combination odierent intrinsic and extrinsic molecular characteristics, which

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Hassimotto et al.

may produce an innite variety o color shades (HARBORNE,1988). Moreover, Guarani cultivar showed the highest rateso cyanidin and total anthocyanins (156 and 194 mg.100 g1,respectively).

Besides cyanidin glycosides, epicatechin was also detectedin concentrations ranging rom 20 to 63 mg.100 g1. HPLC-DADdetected the presence o cyanidin, avan-3-ol epicatechin,quercetin and kaemperol (Figure 1).

According to Wang and Lin (2000), ripe black raspberry andblackberry ruits, on the basis o their wet weight, had higherantioxidant activities than red raspberries or strawberries. Ac-cording to these authors, black raspberry and blackberry containhigh amounts o cyanidin glycosides, a strong antioxidant,whereas strawberries are rich in pelargonidin 3-glucoside andascorbic acid, which are weak antioxidants, due to structuraldierences between the two anthocyanidins.

Te oxidation o-carotene was inhibited in all cultivars,with an inhibition rate ranging rom 66 to 76% (able 3).Tese values are close to those obtained by the syntheticantioxidant butylated hydroxytoluene (BH) (inhibition o

77.6% at 50 M), which characterizes the blackberry ruitsas strong antioxidants. According to these results, Guaranicultivar had the highest antioxidant capacity (inhibition o76 3%) and anthocyanin content (194 5 mg.1001 FW).However, the highest total phenolic and vitamin C contents(499 4 and 21 2 mg.1001 FW, respectively) were ound inCaingangue cultivar.

In this study, a linear correlation was observed betweencyanidin content and antioxidant capacity (r = 0.65). Accord-ing to data by Arabbi, Genovese and Lajolo (2004), the linearcorrelation between avonoids and antioxidant capacity waslower than 0.1. Tis may have happened due to dierences in

Table 1. Physicochemical characteristics o ve Brazilian blackberry cultivars. Dierent letters in columns denote signicant dierences accordingto the ukey test (p < 0.05).

Cultivar Ash (%) Moisture (%) pH SS (Brix) A (% citric acid) Carbohydrates (%) SS/A

Caingangue 0.29 0.006AB 90.88 0.08C 3.42 0.002A 7.60 0.47C 1.26 0.03D 5.04 0.06B 6.03

Brazos 0.31 0.016A 93.25 0.38A 3.31 0.003D 6.19 0.11D 1.54 0.02A 3.07 0.11D 4.01

upy 0.24 0.009D 91.37 0.20B 3.23 0.005F 6.93 0.13CD 1.33 0.02C 4.72 0.17BC 5.21

Guarani 0.29 0.012AB 90.47 0.26C 3.30 0.006D 9.23 0.15B 1.47 0.03B 5.92 0.16A 6.28

Seleo 97 0.25 0.007CD 91.59 0.28B 3.33 0.004C 9.32 0.36AB 1.26 0.01D 4.90 0.10B 7.39n.d.: not detected; and values are means SD (n = 3).

Table 2. otal soluble sugars and soluble sugars (g.100 g1 FW) o ve Brazilian blackberry cultivars. Dierent letters in columns denote signicantdierences according to the ukey test (p < 0.05).

Cultivar Glucose Fructose Sucrose otal soluble sugars

Caingangue 2.44 0.01B 1.91 0.09B n.d. 4.35 0.20C

Brazos 1.69 0.08A 1.15 0.09A n.d. 2.75 0.17A

upy 2.53 0.07B 2.02 0.04B n.d. 4.56 0.11C

Guarani 2.08 0.01B 1.70 0.09B n.d. 3.77 0.22B

Seleo 97 2.60 0.02B 2.11 0.02B 0.7 0.03 4.78 0.35C

n.d.: not detected; and values are means SD (n = 3).

Table 3. otal phenolic, vitamin C and anthocyanin contents (mg.100 g1 FW) and antioxidant capacity (percentage o inhibition) o ve Brazilianblackberry cultivars. Dierent letters in columns denote signicant dierences according to the ukey test (p < 0.05).

Cultivar otal phenolics Antioxidant capacity Vitamin C Anthocyanins

Caingangue 499 4D 71 4A 21.0 2C 125.6 0.8B

Brazos 341 22A 66 2A 9.9 0.7A 133.0 3B

upy 373 17A,B 71 2A 14.0 1B 116.0 2A

Guarani 427 8C 76 3A 11.9 0.9A 194.0 5C

Seleo 97 408 14B,C 72 2A 15.6 0.5B 116.0 2A

Values are means SD.

Table 4. Flavonoid content (mg.100 g1 FW) o ve Brazilian blackberry cultivars, expressed as aglycone. Dierent letters in columns denotesignicant dierences according to the ukey test (p < 0.05).

Cultivar Quercetin Kaemperol Cyanidin Epicatechin otal

Caingangue 19.0 2.0C r 142.0 6.0C 52.0 3.0B 213.29

Brazos 8.0 1.0A r 91.0 2.0A 49.0 5.0B 149.55

upy 7.8 0.3A r 94.9 0.2A 20.7 0.8A 123.29

Guarani 13.3 0.7B r 157.0 3.0C 63.0 2.0C 233.34

Seleo 97 9.0 0.1A r 122.0 1.0B 52.0 3.0B 182.97

Values are means SD; r: trace.

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the composition o phenolics between vegetal extracts and their

reaction with Folin-Ciocalteu reagent (KHKNEN et al.,

1999). Moreover, values o antioxidant capacity may change

according to the methodology chosen.

Flavonoid antioxidant capacity is related mainly to struc-

tural characteristics, such as the presence o an o-diphenil group

in the B ring, the presence o a 3-hydroxyl group connected to a

double bond between C2C

3and adjacent to a 4-oxo unction in

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Figure 1. Chromatographic prole o avonoids rom blackberry cultivars obtained by HPLC-DAD (270 nm). Cyanidin derivative (cy), quercetinderivative (querc), kaempherol derivative (kaemp), epicatechin (epicat).

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CONNOR, A. M. et al. Changes in ruit antioxidant activity amongblueberry cultivars during cold-temperature storage. Journal ofAgricultural and Food Chemistry,v. 50, n. 4, p. 893-89, 2002.

CORDENUNSI, B. R. et al. Eects o temperature on the chemicalcomposition and antioxidant activity o three strawberry cultivars.Food Chemistry, v. 91, n. 1, p. 113-121, 2003.

DEIGHON, N. et al. Antioxidant properties o domesticated and wild

Rubus species. Journal of the Science of Food and Agriculture,v. 80, n. 9, p. 1307-1313, 2000.

DREWNOWSKI, A.; GOMEZ CARNERO, C. Bitter taste,phytonutrients, and the consumer: a review. American Journal ofClinical Nutrition,v. 72, n. 6, p. 1424-1435, 2000.

DUBOIS, M. et al. Colorimetric method or determination o sugarsand related substances. Analytical Chemistry,v. 28, n. 3, p. 350,1956.

HAGIWARA, A.; YOSHINO, H.; ICHIBARA, . Prevention bynatural ood anthocyanins, purple sweet potato color andred cabbage color, o 2-amino-1-metil-6-enilimidazol-[4,5b]piridina associated coorectal carcinogenesis in rats initiated with1,2-dimethylhydrazine. Journal of Toxicology Science, v. F27,n. 1, p. 57-68, 2002.

HARBORNE, J. B. Flavonoids: Advances in research since 1980. NewYork: Chapman and Hall Ltd, 1988.

HASSIMOO, N. M. A.; GENOVESE. M. I.; LAJOLO, F. M.Antioxidant activity o dietary vegetables, ruits and commercialrozen ruits pulp. Journal of Agricultural and Food Chemistry,v. 53, n. 8, p. 2928-2935, 2005.

______. Identication and characterization o anthocyanins rom wildmulberry ( Morus nigra L.) growing in Brazil. Food Science andTechnolology International, v. 13, n. 1, p. 17-25, 2007.

KHKNEN, M. P. et al. Antioxidant activity o plant extractscontaining phenolic compounds. Journal of Agricultural and FoodChemistry,v. 47, n. 10, p. 395-3962, 1999.

KHKNEN, M. P.; HEINONEN, M. Antioxidant activity oanthocyanins and their aglycons. Journal of Agricultural and FoodChemistry,v. 51, n. 3, p. 628-633, 2003.

LEE, S. K.; KADER, A. A. Preharvest and postharvest actors inuencingthe vitamin C content o horticultural crops. Postharvest Biologyand Technology, v. 20, n. 3, p. 207-220, 2000.

LIMA, V. L. A. G. et al. Avaliao do teor de antocianinas em polpa deacerola congelada proveniente de rutos de 12 dierentes aceroleiras(Malpighia emarginata D.C.). Cincia e Tecnologia de Alimentos,v. 23, n. 1, p. 101-103, 2003.

LIMA, V. L. A. G.; MELO, E. A.; LIMA, D. E. S. Eeito da luz e datemperatura de congelamento sobre a estabilidade das antocianinasda pitanga roxa. Cincia e Tecnologia de Alimentos, v. 25, n. 1,

p. 92-94, 2005.PERKINS VEAZIE, P.; COLLINS, J. K. Quality o erect-type blackberry

ruit ater short intervals o controlled atmosphere storage.Postharvest Biology and Technology, v. 25, n. 2, p. 235-239,2002.

PIEA, P. G. Flavonoids as Antioxidants. Journal of NaturalProducts,v. 63, n. 7, p. 1035-1042, 2000.

RICE-EVANS, C.; MILLER, N. J.; PAGANGA, G. Structure-antioxidantactivity relationships o avonoids and phenolic acids. Free RadicalBiology and Medicine,v. 20, n. 7, p. 933-956, 1996.

RIZZOLO, A.; FORNI, E.; POLESELO, A. HPLC assay o ascorbic acidin resh and vegetables. Food Chemistry,v. 14, n. 3, p. 189-199,1984.

the C ring, and the hydroxylation pattern, mainly C5

and C7

inthe A ring (SEERAM; NAIR, 2002). Te absence or replacemento some o these structural characteristics reduces or inhibitsantioxidant capacity. According to Rice-Evans et al. (1996),avonoids showing the highest EAC values (rolox equivalentantioxidant capacity) in the ABS system are: quercetin avonol,which shows all the structural characteristics mentioned above

(4.72 0.10 mM); cyanidin and delphinidin anthocyanidins,which show the same hydroxyl vicinity arrangement o quercetin(4.4 0.1 mM), and catechin esteried to gallic acid (epicatechingallate and epigallocatechin gallate). In this latter case, thehigh EAC values are a reex o the contribution o gallic acid(4.93 0.02 and 4.75 0.06 mM, respectively). In vegetables,avonoids are mainly ound as glycoside derivatives. Becauseo this, changes in antioxidant capacity may be expected inrelation to their aglycones. Apparently, an important criterionto predict a high antioxidant capacity in vegetable oods istheir anthocyanin content. Foods rich in anthocyanins nor-mally show high antioxidant capacity, when compared to thosewithout this class o avonoids (HASSIMOO; GENOVESE;

LAJOLO, 2005). Based on these acts, anthocyanins are believedto be the main phenolic compounds responsible or the highantioxidant capacity shown by blackberry ruit extracts, sincea high correlation between cyanidin content and antioxidantcapacity was observed.

4 Conclusion

Among the ve blackberry cultivars studied, Cainganguecultivar showed the highest vitamin C and total phenolicscontents. However, Guarani cultivar showed the highest valuesor antioxidant capacity, total avonoids, anthocyanins and cya-nidin, in addition to physicochemical characteristics adequate

or in natura consumption. Tese data conrm the generalafrmation that the regular ingestion o blackberry ruits maycontribute to an important intake o antioxidant polyphenols.

Acknowledgments

Tis study was accomplished in collaboration with theEstao Experimental de Caldas (EPAMIG/FECD) and the sup-port o the Conselho Nacional de Desenvolvimento Cientfco eTecnolgico (CNPq), Fundao de Amparo Pesquisa do Estadode So Paulo (FAPESP) and Fundao de Coordenao deApereioamento de Pessoal de Nvel Superior(CAPES). Authorsalso wish to thank Mrcia de Moraes, Lcia Justino da Silva and

Dr. nia Shiga or technical support.

References

ANUNES, L. E. C.; DUARE FILHO, J.; SOUZA, C. M. Conservaops-colheita de rutos de amoreira-preta. Pesquisa AgropecuriaBrasileira, v. 38, n. 3, p. 413-419, 2003.

AOAC. Official methods of analysis. 16 ed. Washington,DC: Association o Ofcial Analytical Chemists, 1995.

ARABBI, P. R.; GENOVESE, M. I; LAJOLO, F. M. Flavonoids invegetable oods commonly consumed in Brazil and estimatedingestion by the Brazilian population. Journal of Agricultural andFood Chemistry,v. 52, n. 5, p. 1124-1131, 2004.

8/7/2019 a29v28n3

7/7



developmental stage. Journal of Agricultural and Food Chemistry,v. 48, n. 2, p. 140-146, 2000.

WU, X.; PRIOR, R. L. Systematic identication and characterizationo anthocyanins by HPLC-ESI-MS/MS in common oods in theUnited States: Fruits and berries. Journal of Agricultural and FoodChemistry,v. 53, n. 7, p. 2589-2599, 2005a.

WU, X.; PRIOR, R. L. Identiication and characterization oanthocyanins by high-perormance liquid chromatography-electrospary ionization-andem mass spectrometry in commonoods in the United States: vegetables, nuts, and grains. Journalof Agricultural and Food Chemistry, v. 53, n. 8, p. 3101-3113,2005b.

ZIELINSLI, H; KOZOLWSKA, H. Antioxidant activity and totalphenolics in selected cereal grains and their dierent morphologicalractions. Journal of Agricultural and Food Chemistry,v. 48, n. 6,p. 2008-2016, 2000.

SEERAM, N. P.; NAIR, M. G. Inhibition o lipid peroxidation andstructure-activity-related studies o the dietary constituents

anthocyanins, anthocyanidins, and catechins. Journal of

Agricultural and Food Chemistry, v. 50, n. 19, p. 5308-5312,

2002.

SEYMOUR, G. B.; AYLOR, J. E.; UCKER, G. A. (Eds.). Biochemistry

of fruit ripening. London: Chapman & Hall, 1996.

SAFFORD, H. A. Flavonoid metabolism. Boca Raton: CRC Press,1990. 298p.

ALL, J. M. et al. art cherry anthocyanins suppress inammation-

induced pain behavior in rat. Behavioral Brain Research, v. 153,

n. 1, p. 181-188, 2004.

WANG, S. Y.; LIN, H. S. Antioxidant activity in ruits and leaves o

blackberry, raspberry, and strawberry varies with cultivar and

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