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Antioxidant compounds and total antioxidant activity in fruits of acerola from cv. Flor Branca, Florida sweet and BRS 366/Compostos antioxidantes e atividade antioxidante total em frutos das variedades Flor Branca, Florida sweet e BRS 366 de aceroleira.


Fruit consumption has been extensively associated to decreases in cardiovascular disease and cancer risks supported by substantial epidemiological evidence (SILVA et al., 2004; AGUDO et al., 2007;). This beneficial effect is mainly due to the presence of antioxidants that neutralize or scavenge reactive species or free radicals thereby, reducing oxidative damage to the cell metabolism. The antioxidant cell defense system is constituted of enzymes that act in concert with non-enzymatic compounds through synergistic or antagonistic interactions contributing to the equilibrium between radical production and elimination.

Previous works showed that acerola presented high levels of both enzymes and nonenzymatic compounds with antioxidant activity (OLIVEIRA et al., 2011; OLIVEIRA et al., 2012), implying therefore, a great potential for human health promotion and for extension of postharvest conservation period. As stated by Lacan and Baccou (1998) when studying melon varieties, those fruit with higher antioxidant levels were able to be stored for longer periods, representing an extension in shelf-life. However, Menichini et al. (2009) reported that quality and quantity of such bioactive compounds are directly influenced by harvest stage, genotype, climate and cultivation techniques.

In Brazil, growers' interest in acerola increased mainly after 1988, when consumers worldwide became aware of its awesome vitamin C levels. Then and initially, the main concern was to establish new varieties which were adapted to the country's northeast region soil and climatic conditions resulting in high yields of quality fruit. Therefore, Embrapa Agroindustria Tropical and others Brazilian research institutions started on breeding programs which resulted in different acerola varieties as Flor Branca and BRS 366, which were launched and recommended for commercial planting by the former institution based on performance of their morphological characteristics, production and fruit quality. Besides those varieties developed in Brazil, others were also imported from abroad as the extraordinarily sweet-flavored 'Florida Sweet' which was established by North-American researchers in Florida and Puerto Rico, EUA.

As other climacteric fruit with a fast ripening metabolism, acerola should be harvested at physiological maturity and may be stored under ambient conditions for only four days or up to 12 days if refrigerated (12[degrees]C) and under PVC film modified storage atmosphere (MOURA et al., 2007). Such short postharvest life, requires that acerola growers have clearly defined which market they will aim at; as for vitamin C and polyphenol industrial extraction where fruit are processed into powder, capsules or concentrated forms for foodstuff supplementation or as for fresh, frozen fruit or pulp consumers. Based on this data and hoping that information on antioxidant properties at different ontological stages will help producers and food technologists to identify which cultivar and/or maturity stage are most adequate for their need, this work aimed to study the changes in the antioxidant metabolites and in antioxidant capacity during development of acerola fruit from cv. Flor Branca, BRS 366 and Florida Sweet.


Fruit material

Acerola clones: Flor Branca, BRS366 and Florida Sweet were obtained from Embrapa Agroindustria Tropical Experimental Field at Pacajus-CE, Brazil, where they were harvested at different stages according to skin color and size as shown in Figure 1: immature fruits with green color (I), physiologically mature with green color and maximum size (II), breaker--turning red (III) and full red ripe (IV). After harvest, fruits were selected based on homogeneity in color/size and absence of defects, washed in tap water and divided in samples that were evaluated as four replications with 500 g each. Fruit mass ranged from 3-8 g. Fruit samples were then, processed using a domestic blender Walita0, the processed pulp was stored at -20[degrees]C and within a 30 day period, was further analyzed as following.


2,6-Dichloro-indophenol (DFI), Folin-Ciocalteu Reagent, Formic acid, Acetonitrile, Anthrone, Cyanidin 3-rhamnoside, Quercetin 3-rhamnoside, Pelargonidin 3-rhamnoside, 2,2-Azinobis-3-ethylbenzthiazoline-6-sulphonic acid radical cation ([ABTS.sup.*+]), 6-Hydroxy-2,5,7,8tetramethylchroman-2-carboxylic acid (Trolox), Ethylenediaminetetracetic acid (EDTA), Nitroblue tetrazolium chloride (NBT) and Hydrogen peroxide ([H.sub.2][O.sub.2]) were purchased from Sigma Chemical Co. (St. Louis, MO).

Quality parameters and non-enzymatic antioxidants

Titratable acidity (TA) of acerola pulp was evaluated as determined by AOAC (2005) using an automatic titrator (Mettler-Toledo[R] DL12, Columbus-USA) and results were expressed as % of malic acid. The pH was measured using an automatic pHmeter (Labmeter[R] PHS-3B, Sao Paulo-Brazil) as recommended by AOAC (2005). Soluble solids (SS) content was determined by refractometry as described by AOAC (2005) using a digital refractometer (ATAGO[R] N1, Kirkland-USA) with automatic temperature compensation. The results were expressed in Brix (concentration of sucrose w/w). The ratio between soluble solids and titratable acidity (SS/TA) was also calculated. Total soluble sugar content was determined by the Anthone method as described by Yemn and Willis (1954) and results were expressed as percentage, %.

The total vitamin C was determined by titration with 0.02% 2,6-dichloro-indophenol (DFI) as method by Strohecker and Henning (1967). One gram of pulp was diluted to 100 mL of 0.5% oxalic acid and homogenized. Then, 5 mL of this solution was diluted to 50 mL with distilled water and titrated and results were expressed as mg.100 [g.sup.-1] FW (fresh weight). Anthocyanins and yellow flavonoids were extracted and determined as described by Francis (1982). One gram of pulp was extracted with a 95% ethanol/1.5 N HCl (85:15) solution, vortexed for 2 min and then, brought to 50 mL with the extracting solution. Protected from the light, the mixture was refrigerated at 4[degrees]C for 12 hours, then filtered on Whatman0 N.1 paper and the filtrate was gathered. The absorbance of the filtrate was measured at 535 nm for the total anthocyanin content using an absorption coefficient of 98.2 mol/cm and at 374 nm for the total yellow flavonoid content using an absorption coefficient of 76.6 mol/cm. Both results were expressed as mg.100 [g.sup.-1] FW.

The total phenol content was measured by a colorimetric assay using Folin-Ciocalteu reagent as described by Ainsworth and Gillespie (2007). Before the colorimetric assay, the samples were subjected to extraction in 50% methanol and 70% acetone as described by Larrauri et al. (1997). Extracts were added to 1 mL Folin Ciocalteau reagent (1 N), 2 mL [N.sub.a2]C[O.sub.3] at 20% and 2 mL of distilled water and absorbance was monitored at 700 nm and the results were calculated from a standard curve of gallic acid 98% (0-50 pg) and expressed as gallic acid equivalents (GAE) mg.100 [g.sup.-1] FW.

Phenols were also determined by liquid chromatography (LC) coupled to mass spectrometer (MS). For extraction, 300 mg of freeze-dried acerola pulp was suspended in 5 mL of 40% methanolic solution, vortex-mixed for 1 min and sonicated for 60 min prior to centrifugation at 2500 g for 10 min at 20[degrees]C. The supernatant was filtered (0.45 [micro]m) and then submitted to chromatographic analysis. The LC DAD-ESI/MS was a Varian[R] ProStar system HPLC (Walnut Creek, CA) coupled with a diode array detector (DAD) and a 500-MS IT mass spectrometer (Varian). A Symmetry[R] C18 column (5.0 [micro]m, 250 x 4.6 mm) was used at a flow rate of 400 [micro]L.[min.sup.-1]. The column oven temperature was set at 30[degrees]C. The mobile phase consisted of a combination of A (0.1% formic acid in milli-Q water) and B (0.1% formic acid in acetonitrile). The gradient varied linearly from 10 to 26% B (v/v) in 40 min, to 65% B at 60 min and finally, to 100% B at 70 min and then, held at 100% B for 75 min. The DAD was set at 340, 270 and 512 nm for real-time read-out and UV/VIS spectra from 190 to 650 nm were continuously collected.

Mass spectra were simultaneously acquired using electrospray ionization in the positive and negative ionization modes (PI and NI) at a fragmentation voltage of 80 V for the mass range of 200-1000 (m/z). A drying gas pressure of 35 psi, nebulizer gas pressure of 40 psi, a drying gas temperature of 370[degrees]C, capillary voltages of 3500 V for PI and NI, and spray shield voltages of600 V were used. The LC system was directly coupled to the MS without stream splitting. Compound identification was primarily based on mass spectrometric data for molecular ions and MS-MS product ions and on published observations for phenolics in fruits and vegetables. Quantification was performed on the basis of UV-Vis data. The UV-visible detector was set to collect the signal at 512 nm for cyanidin 3-rhamnoside and pelargonidin 3-rhamnoside and 340 nm for quercetin 3-rhamnoside. External standard curves were used and results were expressed as equivalents (mg).100 [g.sup.-1] of dry matter (DM).

Total antioxidant activity

The total antioxidant activity (TAA) was determined using 2,2-azinobis-3-ethylbenzthiazoline6-sulphonic acid radical cation ([ABTS.sup.*+]) method as described by Rufino et al. (2006). Before the colorimetric assay, the samples were subjected to a procedure of extraction in 50% methanol and 70% acetone (LARRAURI et al., 1997). Once the radical was formed, the reaction was started by adding 30 [micro]L of extract in 3 mL of radical solution, absorbance was measured (734 nm) after 6 min and the decrease in absorption was used to calculate the total antioxidant activity (TAA). A calibration curve was prepared and different trolox concentrations (standard trolox solutions ranging from 100 to 2000 [micro]M) were also evaluated against the radical. Antioxidant activity was expressed as trolox equivalent antioxidant capacity (), [micro]mol TEAC. [g.sup.-1] FW.

Activity of antioxidant enzymes

Two grams of fruit pulp were homogenized in 10 mL of 0.1 M potassium phosphate buffer (pH 7.0) containing 0.1 mM ethylenediaminetetracetic acid (EDTA) for 1 min, followed by centrifugation at 3248 g for 40 min at 4[degrees]C (YANG et al., 2009). The supernatant fraction was used as a crude extract for the enzyme activity assays and all the procedures were performed at 4[degrees]C. The total protein content was determined according to Bradford (1976) using bovine serum albumin (BSA) as a standard.

Superoxide dismutase (SOD, EC activity was determined spectrophotometrically based on inhibition of the photochemical reduction of nitroblue tetrazolium chloride (NBT) (BEAUCHAMP; FRIDOVICH, 1971; GIANNOPOLITIS; RIES, 1977). The reaction mixture absorbance was measured by the Spectrum SP 2000UV Spectrophotometer at 560 nm and one unit of SOD activity (UA) was defined as the amount of enzyme required to cause a 50% reduction in the NBT photo-reduction rate. Thus, results were expressed as UA.m[g.sup.-1] P (protein).

Catalase (CAT, EC activity was measured according to Beers and Sizer (1952). The reaction started by adding the enzyme extract, then, the decrease in hydrogen peroxide ([H.sub.2][O.sub.2]) was monitored through absorbance at 240 nm and quantified by its molar extinction coefficient (36 M.[cm.sup.-1]). One unit of CAT activity (UA) was defined as the amount of enzyme required to decompose [H.sub.2][O.sub.2] and results were expressed as [micro]mol [H.sub.2][O.sub.2]. [min.sup.-1].[mg.sup.-1] P.

Ascorbate peroxidase (APX, EC activity was assayed according to Nakano and Asada (1981). Enzyme activity was measured using the molar extinction coefficient for ascorbate (2.8 mM.[cm.sup.-1]), considering that 1 mol of ascorbate is required for a reduction of 1 mol [H.sub.2][O.sub.2]. Results expressed as pmol [H.sub.2][O.sub.2].[min.sup.-1].[mg.sup.-1] P.

Statistical analysis

The experimental design was completely randomized in factorial 3 x 4 (cultivars x harvest stage) with four replications with 500 g each. The data obtained was subjected to analysis of variance (ANOVA) using a computer program (SISVAR 3.01) and the averages were compared by the Tukey test at 5% probability (GOMES, 1987).


Quality and non-enzymatic antioxidants

During the ripening (stages III and IV), the soluble solids content was significantly higher and the greatest values were found for 'Florida Sweet' fruit (9.46 Brix), while the lowest were for 'BRS 366' (6.33 Brix) (Table 1). The opposite behavior was observed for titratable acidity which decreased significantly and at stage IV, was lowest in 'Florida Sweet' fruit (0.61%) and highest in 'BRS 366' (1.06%) although, pH values only slightly changed (Table 1). These results may be explained as organic acids are converted into sugars by gluconeogenesis, as observed by the increase in sugar content in Table 1, or are consumed as substrates of respiratory process. Corroborating with the above results, stage IV 'Florida Sweet' fruit presented the highest SS/ TA ratio (15.42) (Table 1). It is noteworthy the exceptionally high sugar content found for ripe 'Florida Sweet' acerola fruit, 6.03%, highlighting its great potential for fresh consumption.

The changes in non-enzymatic antioxidant compounds during acerola development may is shown in Table 2. Vitamin C content decreased more than 40% during ripening when 'BRS 366' fruit showed the greatest values (1363 mg.100 [g.sup.-1]) and 'Florida Sweet', the lowest (862 mg.100 [g.sup.-1]). During development, acerola fruit 'BRS 366' presented higher vitamin C levels, especially at stage I, 2534 mg.100 [g.sup.-1]. Acerolas fruit cv. BRS 366 and Florida Sweet showed contrasting behavior regarding their sugar (Table 1) and vitamin C contents; as the one with highest vitamin level accumulated less sugar ('BRS 366') and the one with the sweetest fruits had a lower vitamin C concentration ('Florida Sweet'). Despite the observed decline, the evaluated acerola varieties still presented high vitamin C contents compared to other fruits as orange, guava and cashew apple with 62.50, 135.70 and 279.37 mg.100 [g.sup.-1], respectively (COUTO; CANNIATTI-BRAZACA, 2010; SILVA et al., 2010; LOPES et al., 2012).

The polyphenol content of 'Florida Sweet' and Flor Branca acerolas fruit decreased in more than 60%, during ripening. 'BRS 366' fruit maintained the highest levels with 4338 and 2631mg GAE.100 [g.sup.-1] at stages I and IV, respectively. The decrease in polyphenols content during ripening reduces fruit astringency leading to a desirable sensory quality and may be explained by polymerization or oxidation by polyphenoloxidase enzyme activity (SHWARTZ et al., 2009). Much though, the phenolic content in acerola is still high when compared to other fruits as papaya with 445 mg GAE.100 [g.sup.-1], pineapple with 298 mg GAE.100 [g.sup.-1], tamarind with 122.2 mg GAE.100 [g.sup.-1] and soursop with 1491 mg GAE.100 [g.sup.-1] (ALMEIDA et al., 2011; SOUSA et al., 2012). The results found for vitamin C and polyphenol content indicates 'BRS 366' acerola fruit as a good source of bioactive compounds to be employed by the nutraceutical industry, especially at physiological maturity (stage I).

Among the polyphenols evaluated in acerola fruits, the anthocyanins and yellow flavonoids are the most abundant and anthocyanin content increased drastically with ripening, especially in 'Flor Branca' fruit reaching 12.37 mg.100 [g.sup.-1], at stage IV A previous work with other acerola varieties showed anthocyanin contents ranging from 0.19 to 47.4 mg.100 g-1 (OLIVEIRA et al., 2012). The main anthocyanin were determined by HPLC-DAD-ESI/MS as two peaks were detected (Figure 2). Peaks 1 and 2 of mass spectrum show the most intense molecular ions m/z 287 and m/z 271 corresponding to cyanidin 3-rhamnoside and pelargonidin 3-rhamnoside, respectively. The latter one was detected only in ripe (stage IV) 'Florida Sweet' and 'Flor Branca' fruits and the greatest levels were observed in ripe 'Flor Branca' fruit, 520.76 mg.100 [g.sup.-1] DM of cyanidin and 97.04 mg.100 [g.sup.-1] DM of pelargonidin, which may be directly related to the total anthocyanin content (Table 2).

According to De Rosso et al. (2008), cyanidin 3-rhamnoside was also the main anthocyanin found in acerola from Waldy Cati 30 and Olivier varieties reaching 78% of its total content. Brito et al (2007) also identified cyanidin 3-rhamnoside (70%) and pelargonidin 3-rhamnoside (30%) as the main anthocyanins in acerola. Thus, it is possible to concluded the sharp increase in the anthocyanin levels makes it the most important phenol present in ripe acerola (stage IV), as was also stated by Hanamura, Uchida and Aoki (2008).

Unlike anthocyanins, yellow flavonoid content increase slightly during ripening (stage IV) being highest in 'Flor Branca' fruit (9.82 mg. 100 [g.sup.-1]). Thereby, acerola cv. Flor Branca stands out in terms of flavonoids, anthocyanins and vitamin C contents which together must influence the soluble solids content found in stage IV fruit (Table 1), since the sugar levels were low (Table 1).

In all acerola varieties, the most abundant yellow flavonoid identified was quercetin 3-rhamnoside, taking into account the molecular ion at m/z 303 (Figure 3). Ripe stage IV acerola showed the highest levels of quercetin, especially for 'Flor Branca' and 'BRS 366' fruit with 33.72 and 27.81 mg.100 [g.sup.-1] DM, respectively. Oliveira et al (2012) found similar contents of quercetin in ripe 'BRS 237' acerola fruits, 33.49 mg.100[g.sup.-1] DM.

The yellow flavonoid quercetin is a powerful antioxidant accumulated during ripening in acerola, adding to its nutritional bioactive properties. According to Hanamura, Kawagishi and Hagiwara (2005), the cyanidin-3-rhamnoside showed a strong neutralizing capacity of the superoxide radical ([O.sub.2.sup.-]), similar to that presented by quercetin and the authors explained that antioxidant activity is strongly correlated the number of B-ring hydroxyls of the structure of polyphenols. However and despite quercetin being present throughout acerola fruit development, its content was much lower when compared to cyanidin in stage IV fruits explaining the red color characteristic of ripe acerola fruit.

Antioxidant enzyme activity

Activity of all antioxidant enzymes evaluated in acerola pulp fruit decreased with development, but especially at ripening between III to IV (Table 3). The activity of SOD decreased significantly, but more drastically in 'Flor Branca' fruit from 3330.50 in stage I to 373.09 UA.[mg.sup.-1]P in stage IV. SOD belongs to a class of metalloenzymes that catalyze the degradation of superoxide anion ([O.sub.2.sup.-]) into [H.sub.2][O.sub.2] and oxygen ([O.sub.2]) and it is considered the first line of defense against reactive oxygen species (ROS) (HUANG et al., 2007).

In plant cells, the second defense line of the antioxidant enzymatic system is constituted of both CAT and APX which are involved with [H.sub.2][O.sub.2] scavenging, however APX uses ascorbate as the electron donor for [H.sub.2][O.sub.2] neutralization. CAT activity was the highest among the evaluated enzymes for the three acerola fruit varieties and it also declined significantly with development. Acerola fruit cv. Flor Branca presented the most drastic reduction in CAT activity, over 90% reaching 555.57 gmol [H.sub.2][O.sub.2].[min.sup.-1]. [mg.sup.-1] P. As for the other two evaluated enzymes, APX activity was higher in 'Flor Branca' fruit clone which decreased from 26.04 in stage I to 5.2 [micro]mol [H.sub.2][O.sub.2]. [min.sup.-1].[mg.sup.-1]P, in stage IV APX activity was much lower than CAT and as both enzymes use [H.sub.2][O.sub.2] as substrate, this result implies that CAT is possibly the main [H.sub.2][O.sub.2] scavenger in acerola fruit, which was also reported by Oliveira et al (2012).

The enzymatic antioxidant defense system present is responsible for the removal of free radicals or ROS leading to protection against oxidative stress and therefore, increasing its postharvest conservation potential. Wang and Chen (2010) reported that fruits with better antioxidant enzyme systems showed a reduction in cell membrane damage which was related to longer periods of postharvest conservation due to a delay in senescence. Thereby, the data here presented indicates that 'Flor Branca' acerola fruit has a greater potential for postharvest conservation due to its higher antioxidant enzymatic activity.

Total antioxidant activity

TAA was significantly reduced for the three acerolas fruit evaluated during development (Table 3), but the steepest fall was found after physiological maturity from stage I to II. In ripe (stage IV) acerola fruit, 'BRS 366' showed the highest TAA with 42.36 [micro]M TEAC.[g.sup.-1]FW, which can be justified by its high vitamin C and polyphenols contents (Table 2). However, variables may contribute differently to AAT as observed for 'Flor Branca' acerola fruit, which showed high contents of all phenolics evaluated although, comparatively low levels of vitamin C, but still presented a high TAA.

Oliveira et al (2011) studied the correlation between antioxidant compounds and TAA and reported that for acerola clone II 47/1, TAA was strongly correlated to SS, as well as polyphenols and vitamin C contents. However, for 'BRS 235' acerola fruit, TAA and vitamin C were negatively correlated with SS, polyphenols and anthocyanins. These authors proposed a compensation system among antioxidants of non-enzymatic and enzymatic nature in acerola fruit, so those varieties with higher antioxidant enzyme activity showed lower nonenzymatic antioxidant levels and vice-versa.





Ripe 'Florida Sweet' acerola fruit presented an exceptionally high sugar content highlighting its potential for fresh consumption. Polyphenol and vitamin C contents decreased more than 40% during ripening, however and despite of the observed decline, the evaluated acerola still presented high contents of such bioactive compounds. Based on the results found for bioactive compounds and for total antioxidant activity, physiologically mature 'BRS 366' acerola fruit seems the best option for the bioactive compounds processing industry. Among the polyphenols evaluated in acerola fruit, cyanidin 3-rhamnoside was the main anthocyanin and quercetin 3-rhamnoside was the most abundant yellow flavonoid. Activity of all evaluated antioxidant enzymes decreased especially at ripening, although 'Flor Branca' acerola fruit kept the highest activity levels, indicating a greater potential for postharvest conservation which, therefore, may be targeted for further studies on postharvest storage. APX activity was much lower than CAT which implies that the latter enzyme is possibly the main [H.sub.2][O.sub.2] scavenger in acerola fruit. Finally, it can be concluded that with ripening, the antioxidant metabolism of acerola fruit is suppressed due to the reduction in vitamin C, in total phenolics and in activity of antioxidant enzymes reflecting in a decrease of total antioxidant activity.


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(1) Trabalho 410-13). Recebido em: 16-10-2013. Aceito para publicacao em: 05-02-2014.

(2) Mestre em Fitotecnia, UFC, Av. Mr. Hull 2297, Campus do Pici, Fortaleza-CE, Brasil. E-

(3) Pesquisado, Embrapa Agroindustria Tropical, Fortaleza-CE, Brasil. E-mails:,

(4) Prof. Adjunta, UFC, Av. Mr. Hull 2297, Campus do Pici, Fortaleza-CE, Brasil; E-mail:
TABLE 1--Changes in postharvest quality during development of
different acerola fruit varieties. Pacajus-CE, 2011.

CV         Stage      SS        TA (%     SS/TA    Sugars      pH
                   (Brix)       malic                (%)

'Florida     I       7.06Ab     0.90Ba   7.80Ab    3.59Ab    3.55Ac
Sweet'      II       7.76Bb     0.84Ba   9.24Bb    3.77Ab    3.56Ac
            III      9.10Cc     0.83Ba   10.96Cb   5.72Bb    3.56Ab
            IV       9.46Cc     0.61Aa   15.42Db   6.03Bb    3.68Bc
'Flor        I       5.50Aa     1.15Bb   4.78Aa    1.60Aa    3.29ABb
Branca'     II       5.63Aa     1.14Bb   4.95Aa    1.55Aa    3.27ABb
            III      6.40Bb     1.19Bb   5.39Aa    2.47Ba    3.23Aa
            IV       6.76Bb     0.97Ab   6.99Ba    2.75Ba    3.32Bb
             I       6.66Cb     1.46Cc   4.56Aa    2.11ABa   3.19Aa
'BRS 366'   II      5.96ABa     1.24Bc   4.80ABa   1.72Aa    3.17Aa
            III      5.46Aa     1.17Bb   4.67Aa    2.48Ba    3.20Aa
            IV      6.33BCa     1.06Ac   5.98BCa   2.32Ba    3.18Aa

For each variable, different UPPERCASE letter indicates statistical
difference at p<0.05 between stages and lowercase letter indicates
statistical difference at p <0.05 between clones, according to
Tukey's test.

TABLE 2--Changes in antioxidants compounds during development
of different acerola fruit varieties. Pacajus-CE, 2011.

CV         Stage     Total      Total Phenols   Anthocyanin
                   Vitamin C    (mg GAE.100      (mg. 100
                    (mg. 100     [g.sup.1])     [g.sup.-1])

             I     1501.17Ca      4019.42Ca       2.29Aa
Florida     II     1273.31Ba      2124.11Ba       2.40ABa
Sweet'      III    1239.60Ba      2126.93Ba       2.85Ba
            IV      862.86Aa      1740.75Ab       6.34Cb
             I     1966.44Cb      4338.89Db       2.55Aab
Flor        II     1672.85Bb      2539.3Cb        2.49Aa
Branca      III    1629.91Bc      2010.79Ba       3.23Ba
            IV     1104.57Ab      1561.67Aa       12.37Cc
             I     2534.06Cc      4203.72Db       2.78Ab
BRS 366'    II     1799.46Bc      3279.51Cc       2.41Aa
            III    1328.40Ab      2465.18Ab       3.32Ba
            IV     1363.70Ac      2631.34BC       5.07Ca

CV         Cyanidin   Pelargonidin     Yellow        Quercetin
                                     Flavonoids       (mg.lOO
           (mg. 100 [g.sup.-1] DW)    (mg. 100     [g.sup.-1] DW)

            0.49Aa         nd          7.26Ba         10.44Aa
Florida     9.47Aa         nd          5.99Aa          4.26Aa
Sweet'     44.87Ba         nd          6.01Aa          5.23Aa
           198.52Ca      15.66a        7.36Ba         12.10Aa
            1.07Aa         nd          8.17Bb         16.73Aab
Flor       21.63Aa         nd          7.05Ab         12.73Ab
Branca     111.84Bb      2.26A         6.62Aab        ll.llAab
           520.76Cb     97.04Bb        9.82Cc         33.72Bb
            1.15Aa         nd          8.08Bb         22.5 lAb
BRS 366'   17.98Aa         nd          6.99Ab         16.34Ab
           97.38Bb         nd          7.28Ab         21.50Ab
           143.65Ca        nd          8.08Bb         27.81Ab

For each variable, different UPPERCASE letter indicates statistical
difference at p<0.05 between stages and lowercase letter indicates
statistical difference at p <0.05 between clones, according to
Tukey's test.

TABLE 3--Changes in activity of antioxidant enzymes and in total
antioxidant activity (TAA) during development of different
acerola fruit varieties. Pacajus-CE, 2011

                     SOD      CAT               APX         TAA
          Stage   (UA.[mg.      [micro]mol [H.sub.2]     ([micro]M
                  sup.-1]     [O.sub.2], .[min.sup.-1].    TEAC.
                     P)           [mg.sup.-1] P)         [g.sup.-1]

Florida     I     2027.05Ca   7255.66Cb       23.73Cb     104.96Ca
Sweet      II     1261.16Ba   3535.07Bb       15.11Bb     36.28Ba
           III    1053.06Ba   2563.50Bb       29.76Cb     42.64ABa
           IV     532.09Ac    844.97Aa        4.75Aab     32.93Aa
Flor        I     3330.50Cb   9544.43Cc       26.04Bb     120.93Ca
Branca     II     1147.89Bb   2336.81Ba       2.94Aa      49.90Bb
           III    596.62Aa    915.57Aa        3.48Aa      46.17ABa
           IV     373.09Aa    555.57Aa        5.20Aa      39.20Aab
BRS 366     I     1133.82Bb   5504.80Ca      11.35ABa     140,04Ca
           II     1116.62Bc   3289.81Bb       18.32Bb     75.33BC
           III    750.44Ab    1915.19Ab       13.59Bb     41.04Aa
           IV     519.53Ab    938.23Aa        8.04Ab      42.36Ab

For each variable, different UPPERCASE letter indicates statistical
difference at p<0.05 between stages and lowercase letter indicates
statistical difference at p <0.05 between clones, according to
Tukey's test.
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