Haminiuk 2011

International Journal of Food Science and Technology 2011, 46, 1529–1537 1529
Original article
Chemical, antioxidant and antibacterial study of Brazilian fruits
Charles Windson Isidoro Haminiuk,1* Manuel Salvador Vicente Plata-Oviedo,1 Amanda Roman Guedes,1 Ana Paula
Stafussa,1 Evandro Bona1 & Solange Teresinha Carpes2
1 Universidade Tecnológica Federal do Paraná, Programa de Pós-Graduação em Tecnologia de Alimentos (PPGTA), Campus Campo Mourão,
PR, Brazil
2 Universidade Tecnológica Federal do Paraná, Campus Pato Branco, PR, Brazil
(Received 18 October 2010; Accepted in revised form 31 March 2011)
Summary In this study, the antioxidant capability, total phenolic content and antimicrobial activity of ethanolic
extracts of seven fruits from the Brazilian Atlantic Forest were evaluated. The conditions for the extraction
of crude phenolics from the fruits were determined using an experimental factorial design. Total phenolic
content, 1,1-diphenyl-2-picrylhydrazyl radical (DPPH•) scavenging activity and b-carotene-linoleic acid
couple oxidation assays were used to evaluate the antioxidant properties of the extracts. In addition,
antimicrobial activity was screened using two Gram-negative bacteria (Escherichia coli and Klebsiella
pneumoniae) and one Gram-positive bacterium (Staphylococcus aureus). All native fruits assayed in this study
have high potential as natural antioxidant sources. Among the seven fruits evaluated, Jabuticaba and Uvaia
had the highest antioxidant activity in the DPPH• and of b-carotene-linoleic acid coupled oxidation assays.
In the biological assay, K. pneumoniae was the most sensitive microorganism to the fruit extracts, and the
Jabuticaba extract had a slight inhibitory effect against this Gram-positive bacterium.
Keywords Antimicrobial activity, 1,1-diphenyl-2-picrylhydrazyl radical, extraction, fruit extracts, phenolic compounds.
These fruits represent an opportunity for local produc-

Introduction
ers to gain access to special markets where consumers
Epidemiological studies have shown that dietary patterns place emphasis on exotic character and the presence of
are significantly associated with the prevention of non- nutrients capable of preventing degenerative diseases.
transmissible chronic diseases, such as heart disease, Fruit consumption is not only a matter of taste and
cancer, diabetes and Alzheimer’s disease. Consumption personal preference, but it has also become a concern of
of fruits and vegetables has been highly associated with health because of content of vital fruit nutrients (Alves
the reduced risk of cancer (Sun et al., 2002). In fruits, et al., 2008). Interest in PC has increased dramatically
some of the components involved in the prevention of over recent years because they are present in all plants
degenerative diseases include soluble dietary fibre, insol- and, therefore, are part of human diets (Shahidi &
uble dietary fibre, vitamin C, vitamin E, folate, carote- Naczk, 2004). The PC widely distributed in medicinal
noids, anthocyanins, selenium and phenolic compounds plants, spices, vegetables, fruits, grains, pulses and other
(PC). The combination of vitamins, minerals, phenolic seeds are an important group of natural antioxidants
antioxidants and fibre is responsible for these effects with possible beneficial effects on human health. These
(Ruxton et al., 2006). Moreover, the consumption of compounds can help protect against the harmful action
tropical or exotic fruits has increased worldwide. Trop- of reactive oxygen species, especially oxygen-free radi-
ical fruit consumption is increasing in the domestic and cals (Stratil et al., 2007).
international markets due to growing recognition of its Nevertheless, evaluating antioxidants in food is still
nutritional and therapeutic value. challenging from an analytical point of view because
Brazil has a large number of underexploited native they are always present as complex mixtures. The
and exotic fruit species of potential interest to the bioactivity of antioxidants cannot be attributed to only
agroindustry, which may be a possible future source of one compound or a group of compounds. In addition,
income for the local population (Alves et al., 2008). the absorption, metabolism and physiological effects of
antioxidants are different when they are ingested
*Correspondent: E-mail: [email protected] through the consumption of fruits, vegetables and other
doi:10.1111/j.1365-2621.2011.02653.x
2011 The Authors. International Journal of Food Science and Technology 2011 Institute of Food Science and Technology
13652621, 2011, 7, Downloaded from https://ifst.onlinelibrary.wiley.com/doi/10.1111/j.1365-2621.2011.02653.x by Universidade Federal Do ABC, Wiley Online Library on [08/10/2024]. See the Terms and Conditions (https://onlinelibrary.wiley.com/terms-and-conditions) on Wiley Online Library for rules of use; OA articles are governed by the applicable Creative Commons License
1530 Bioactive compounds of Brazilian fruits C. W. I. Haminiuk et al.
food sources or as supplements. Methodologies for other fruits is not presented in this study due to unnec-
complete extraction of phenolics have not been achieved essary repetition and space restrictions.
yet. PCs are less potent than pharmaceutical drugs, but
the advantage of PC is that they are always present in Phenolic compound extraction
human diets, resulting in a long-term physiological effect The total phenolic compound extraction was carried
(Vasco, 2009). Therefore, with the purpose of identify- out based on the extraction design described in the
ing potential health-promoting effects of unexplored extraction conditions section. The fruits were thawed,
fruits from the Brazilian Atlantic Forest, the main and 10 g of each fruit was crushed, mixed with 40 mL
objective of this study was to investigate several native of 40% ethanol and agitated in a test tube rotary mixer
fruits by analysing their antioxidant and antimicrobial for 1 h. The samples were then centrifuged at 3500 g. for
potential. 10 min. The supernatant (extract) was used to quantify
the total PC and flavonoids, and it was used to test the
antimicrobial activity of the samples.
Materials and methods
Plant material Phenolic compound quantification

In this study, the following seven fruits from the Atlantic The phenolic compound content in the fruit extracts was
Forest (Southern Brazil) were studied: Araçá do campo estimated by a colorimetric assay following the meth-
(Psidium guineense), Cambuci (Campomanesia phaea), odology of Singleton & Rossi (1965). The Folin–
Feijoa (Feijoa sellowiana), Gabiroba (Campomanesia Ciocalteau method was used with Gallic acid as a
pubescens), Grumixama (Eugenia brasiliensis), Jabuti- standard. Properly diluted samples or blanks (100 lL)
caba (Myrciaria cauliflora) and Uvaia (Eugenia uvalha). were pipetted into separate 10 mL volumetric balloons
The fruits were provided by Sitio do Belo (Paraibuna, SP, with 5 mL of distilled water. Folin–Ciocalteau reagent
Brazil), and they were frozen at )18 C in a conventional (500 lL) was then added to the mixture. After 3 min,
home freezer for a maximum period of 1 month. 1.5 mL of 15% sodium carbonate was added, and
distilled water was added to a final volume of 10 mL.
The mixture was incubated in the dark at room
Extraction conditions
temperature for 2 h. The absorbance was then measured
The initial step of the preliminary experiment was to at 765 nm using an UV ⁄ Vis double beam spectropho-
select an appropriate solvent to extract the PC from the tometer T-80 (PG Instruments Limited, Beijing, China).
fruits. Three different solvents were used such as The results were expressed as gallic acid equivalents
ethanol, methanol and distilled water. PC were extracted (GAE) using a calibration curve over the range of
using a series of extraction media with varying percent- 5–250 ppm (Vasco et al., 2008).
ages ranging from 0% to 100% (v ⁄ v; water ⁄ ethanol,
methanol and pure distilled water; Liyana-Pathirana &
Anthocyanins and flavonoids
Shahidi, 2005). In this initial test, 40% ethanol with a
solute ⁄ solvent ratio of 1:4 (w ⁄ v) was the best for all The total anthocyanin content of the fruits was deter-
fruits (data not shown). A full factorial design (3 · 3) mined using the pH differential method (Giusti &
with one block and nine runs was used to evaluate the Wrolsted, 2001). The fruit extracts were separately
influence of time and solute ⁄ solvent ratio for the
extraction of PC from the seven fruits. Experimental Table 1 Full factorial design for phenolic compounds extraction of
data were fitted to a second-order polynomial model, Jabuticaba
and regression coefficients were then obtained. The
Time Volume mg g)1
generalised second-order polynomial model used in the
Experiment (min) (mL) mg L)1* (dry basis)*
response surface analysis was as follows:
X
2 X
2 X
2 1 60 20 472.90 6.86 ± 0.01
Y ¼ b0 þ bi Xi þ bii X2i þ bij Xi Xj ð1Þ 2 60 30 368.40 8.02 ± 0.02
3 60 40 307.90 8.94 ± 0.01
i¼1 i¼1 i<j¼1
4 120 20 465.40 6.76 ± 0.05
where bo, bi, bii and bij are regression coefficients 5 120 30 363.40 7.91 ± 0.02
for intercept, linear, quadratic and interactions terms, 6 120 40 305.40 8.87 ± 0.02
respectively, Xi and Xj are independent variables (Liyana- 7 180 20 466.40 6.77 ± 0.05
8 180 30 361.90 7.88 ± 0.03
Pathirana & Shahidi, 2005). Table 1 shows the experi-
9 180 40 314.90 9.14 ± 0.03
mental design for Jabuticaba fruit. The best conditions
obtained for Jabuticaba fruit were also observed in the *Amount of total phenolic compounds. Moisture content of Jabuticaba
other fruits analysed. Therefore, the extraction data of the (86.23%).
International Journal of Food Science and Technology 2011 2011 The Authors
International Journal of Food Science and Technology 2011 Institute of Food Science and Technology
Bioactive compounds of Brazilian fruits C. W. I. Haminiuk et al. 1531
dissolved in potassium chloride (0.025 mol L)1; pH 1.0) where Abssample is the absorbance of the sample,
and sodium acetate (0.4 mol L)1; pH 4.5) buffers at a Absblank is the absorbance of the blank, and Abscontrol
predetermined dilution factor. The absorbance (A) at is the absorbance of the control. Ethanol (1.0 mL)
520 and 700 nm was determined in a UV ⁄ Vis double mixed with the plant extract solution (2.5 mL) was used
beam spectrophotometer T-80 (PG Instruments Lim- as a blank. The 0.3 mmol L)1 DPPH• solution (1.0 mL)
ited), and the results were calculated as follows: mixed with ethanol (2.5 mL) was used as a negative
A ¼ ðA520 A700 ÞpH 1:0 ðA520 A700 ÞpH 4:5 ð2Þ control. The positive controls included the samples
using the standard solutions. The EC50 values were
The monomeric anthocyanin (MA) pigment concen- calculated by linear regression of plots where the
tration in the sample was calculated as follows: abscissa represented the concentration of tested plant
extracts, and the ordinate was the average percent of
A M DF 1000 antioxidant activity from three separate tests (Sim & Sil,
MA ¼ ð3Þ
ek 2008). Trolox, BHT and propyl gallate were used as
where M is the molar mass of cyanidin-3-glucoside standards with a concentration of 100 ppm.
(449.2 g mol)1), DF is the dilution factor (4.0), e is the
molar extinction coefficient (26 900 L mol)1 cm)1), and Coupled oxidation of b-carotene and linoleic acid
k is the cuvette optical path length (1 cm). The final
anthocyanin concentration was expressed as milligrams The antioxidant activity of the fruit extracts according to
of cyanidin-3-glucoside per litre of extract. The final b-carotene-linoleic acid coupled oxidation assay was
results were expressed in milligrams per gram of fresh measured using the methodology of Emmons et al.
weight (fw). All analyses were done in triplicate. (1999) with modifications proposed by Prado (2009).
The total flavonoid content was determined using the b-Carotene (10 mg) was weighed and dissolved in 100 mL
methodology proposed by Chang et al. (2002). Aliquots of chloroform. Subsequently, 40 mg of linoleic acid
of fruit samples (0.1 g) were dissolved in 1 mL of and 400 mg of Tween 40 were added to 3 mL of
deionised water. This solution (0.5 mL) was then mixed the b-carotene ⁄ chloroform solution. Chloroform was
with 1.5 ml of 95% ethanol, 0.1 mL of 10% aluminium removed under a stream of nitrogen gas. Oxygenated
chloride hexahydrate, 0.1 mL of 1 mol L)1 potassium deionised water (100 mL) was added and mixed well.
acetate, and 2.8 mL of deionised water. After incubation Aliquots of the b-carotene-linoleic acid emulsion (3 mL)
at room temperature for 40 min, the absorbance of the were mixed with 50 lL of the fruit extract (100 ppm), and
reaction mixture was measured at 415 nm against a the mixtures were incubated in a water bath at 50 C.
deionised water blank on a spectrophotometer (UV ⁄ Vis Oxidation of the emulsion was monitored
double beam spectrophotometer T-80; PG Instruments spectrophotometrically using a UV ⁄ Vis double beam
Limited). Quercetin was chosen as a standard. The spectrophotometer T-80 (PG Instruments Limited) by
standard curve was obtained in a concentration range of measuring absorbance at 470 nm over a period of
0–50 mg L)1, and the levels of total flavonoids in fruits 120 min. Control samples contained 50 lL of ethanol
were determined in triplicate. instead of the extract. The degradation over time was non-
linear. Therefore, the antioxidant activity was expressed
as percent inhibition relative to the control after incuba-
1,1-Diphenyl-2-picrylhydrazyl radical (DPPH•) assay tion for 120 min using the following equation:
The free radical scavenging activity was assessed with
DRc DRS
the DPPH• method as previously described by Mensor AOA ¼ 100 ð5Þ
et al. (2001). Based on the total phenolic compound DRc
values, six different concentrations (5, 10, 25, 50, 125
and 250 ppm in ethanol) of the extract were used to where AOA is the antioxidant activity, DRC is the
perform the DPPH• assay. The 0.3 mmol L)1 DPPH• degradation rate of the control (ln(a ⁄ b) ⁄ 120), DRS is
ethanol solution (1 mL) was added to 2.5 mL of sample the degradation rate of the sample (ln(a ⁄ b) ⁄ 120), a is the
solutions of different concentrations, and the mixtures initial absorbance at time zero, and b is the absorbance
were allowed to react at room temperature. After at 120 min. Trolox, BHT and propyl gallate were used
30 min, the absorbance values were measured at as standards with a concentration of 100 ppm.
518 nm, and they were converted into the antioxidant
activity percentage (AA%) using the following equation: Antibacterial activity of the fruit extracts
The disc diffusion method was employed to determine
the fruit extract antibacterial activity. The following
ðAbssample Absblank Þ 100
AA% ¼ 100 ð4Þ bacterial strains were used as test organisms: E. coli
Abscontrol ATCC 25922, S. aureus 25923 and K. pneumoniae
ATCC13883. Paper discs (diameter of 6 mm) were
2011 The Authors International Journal of Food Science and Technology 2011
impregnated with 20 lL of the fruit extract dissolved in gomery, 2001). Figure 1 shows that only the volume
40% ethanol (final concentration of 300 ppm) and had an important role in the phenolic extraction of
transferred onto pretreated Mueller-Hinton agar in Jabuticaba fruit (P £ 0.05), which indicated that the
Petri dishes. The pretreatment consisted of a surface extraction of PC was not dependent on time for the
spread with 0.1 mL of logarithmic phase bacteria at a range studied. Therefore, the smallest length of time
density adjusted to a 0.5 McFarland turbidity standard (60 min) was chosen in this study to extract the PC as
(108 CFU mL)1). Chloramphenicol (1000 mg L)1) was time had no influence on this process. A similar result
used as a positive control, and the negative control was has been reported by Luthria (2008) who studied the
ethanol. After incubation at 37 ± 1 C for 24 h, the influence of experimental conditions on the extraction
diameters of the inhibition zones were measured in of PC from parsley. This author found no significant
millimetres. The tests were carried out in duplicate. changes in the extraction yields when testing three
different static time settings. Thus, an extraction over a
long period may cause the degradation of PC. The two
Statistical analysis
most important factors that facilitate degradation
Assays were performed in triplicate for each sample. The reactions are light and oxygen in the air. Enzymes
results were expressed as the mean values ± SD. (mainly oxidative enzymes) already present in the
Student’s t-test was used for comparison between two fruits, which are released during the extraction, can
means, and a one-way anova was used for comparison promote such degradation (Palma et al., 2001). anova
of more than two means. A difference was considered and the regression coefficients of time and volume
statistically significant when P £ 0.05. The statistical parameters are shown in Tables 2 and 3. In general,
analysis was carried out using statistica 7.1 software proceeding with exploration and optimisation of a
(StatSoft, Tulsa, OK, USA). Error term and model fitted response surface may produce poor or misleading
significance were used to judge the adequacy of model results unless the model exhibits an adequate fit
fitness. (Liyana-Pathirana & Shahidi, 2005). The quality of
the fit of the polynomial model equation was expressed
by the coefficient of determination (R2), and the
Results and Discussion
statistical significance was verified by an F-test. anova
(F-test) showed that the second-order model fitted well
Extraction of phenolic compounds
into the experimental data. The coefficient of determi-
In general, the efficiency of the extraction of a nation (R2) value of the model was 0.9972, and the
compound is influenced by multiple parameters, such adjusted R2 was 0.9926, which indicated that the model
as temperature, time and solvent polarity, and their adequately represented the real relationship between
effects may either be independent or interactive (Mont- the parameters chosen.
Figure 1 Volume vs. time of extraction of

phenolic compounds from Jabuticaba.
Table 2 anova of total phenolic compounds extraction of Jabuticaba products, the taste is heavily influenced by the presence
of phenolics. Moreover, fruits and vegetables represent
Sum of Degrees of Mean
excellent sources of bioactive compounds, and the
Variables squares freedom square F P
consumption of fruits and vegetables has been associ-
Time L + Q 0.0168 2 0.008 1.273 0.397 ated with a reduced risk of many common chronic
Volume L + Q 7.178 2 3.589 541.754 0.0001* diseases, including cancer, cardiovascular disease, and
Time · Volume 0.022 1 0.022 3.345 0.164 chronic inflammatory diseases (Genovese et al., 2008).
Error 0.019 3 0.007 Therefore, an assessment of phenolic content in food is
Total 7.237 8 of great importance (Waterhouse, 2002). In our study,
*P £ 0.05, L, linear; Q, quadratic. Coefficient of determination
seven different fruits from the Brazilian Atlantic Forest
(R2) = 0.9972. were studied for their antioxidant potential. Several of
the fruits selected for the present study are unknown
outside their natural range, and the other fruits are
barely known (Vasco et al., 2008). Table 4 summarises
Table 3 Regression coefficients of second-order polynomial model for
phenolic compounds extraction of Jabuticaba
the contents of PC, flavonoids and anthocyanins found
in Uvaia, Araçá, Jabuticaba, Gabiroba, Feijoa, Gru-
Term Coefficients SE P-value mixama and Cambuci. A wide variation was observed in
the PC contents found in the fruits with the highest and
b0 7.910 0.027 0.000* lowest quantities found in Cambuci (107.69 ± 6.19 mg
Linear
GAE g)1 dry weight) and Jabuticaba (9.55 ± 0.17 mg
)0.005
b1 – Time 0.033 0.880
GAE g)1 dry weight), respectively. Statistical differences
b2 – Volume 1.093 0.033 0.000*
Quadratic
(P £ 0.05) were found among the PC values of the
b11 – Time )0.045 0.028 0.210 samples. Gonçalves et al. (2010) evaluated the antioxi-
b22 – Volume 0.024 0.028 0.462 dant potential of native Brazilian fruits, and they
Cross product observed a high amount of PC in fresh and commer-
b12 0.074 0.041 0.164 cially prepared fruit pulp of Cambuci. Reynertson et al.
(2008) reported that freeze-dried Jabuticaba has a
*P £ 0.05.
phenolic content of 31.6 ± 0.39 mg GAE g)1 dry
weight. Numerous reducing compounds may interfere
in the quantification of polyphenols using the FC
Phenolic compound, flavonoid and anthocyanin content
method. Among them, vitamin C has been suggested
There are many phenolic substances in plants and, to be the major contributor of interference. According
consequently, in foods. Rich dietary sources of pheno- to Georgé et al. (2005), the contribution of vitamin C is
lics include fruits, tea, coffee and cocoa, in addition to 32% of the total interference for orange juice and 46%
processed foods derived from these sources, such as for apple juice. The contribution of vitamin C is
wine. At high levels, phenols impart astringency, bitter- remarkably low in tomato juice, with only 9% of the
ness, and colour to foods, particularly when sugar levels total interference expressed as GAE. There was almost
are low. In red wine, unsweetened tea, and chocolate no available data regarding PC for the majority of the
Table 4 Total phenolics, flavonoids and

anthocyanins content of the fruits* Content of phenolic Content of phenolic Flavonoids Anthocyanins
Fruits compounds† compoundsà (mg 100 g)1 fw§) (mg 100 g)1 fw–)
Uvaia 373.40 ± 14.10e 24.09 ± 0.91c 58.72 ± 2.67b 4.77 ± 0.41e

Araçá 684.73 ± 5.03d 12.82 ± 0.09d 49.46 ± 2.84b 5.37 ± 0.24e
Jabuticaba 312.73 ± 5.77e 9.55 ± 0.17d 31.60 ± 4.00c 342.23 ± 2.31a
Gabiroba 2714.00 ± 216.56b 48.42 ± 3.86b 56.21 ± 3.29b 15.21 ± 0.63d
Feijoa 1834.00 ± 103.92c 54.42 ± 3.08b 77.97 ± 6.25a 70.24 ± 1.20c
Grumixama 568.73 ± 31.50d 25.98 ± 1.43c 14.87 ± 1.53d 266.34 ± 2.92b
Cambuci 3414.00 ± 190.78a 107.69 ± 6.19a 30.16 ± 2.08c 19.44 ± 0.23d
*
Mean value ± SD; n = 3.
†
mg GAE L)1 (milligram of gallic acid equivalents per litre).
à
mg GAE g)1 dw (milligram of gallic acid equivalents per gram dry weight).
§
(milligrams of quercetin per 100 g of fresh weight).
–
(milligrams per 100 g of fresh weight). Values in a column followed by the same letter are not
significantly different (P > 0.05).
fruits investigated in this study, which made it difficult antioxidant behaviour of Brazilian fruits has been
to perform a data comparison. The content of total investigated by Rufino et al. (2010b), and they observed
flavonoids varied from 14.87 ± 1.53 to 77.97 ± 6.25 a total anthocyanin content of 58.1 ± 0.9 mg 100 g)1
mg quercetin 100 g)1 fw. These amounts were compa- fw in Jabuticaba. Nevertheless, Santos et al. (2010)
rable with results previously described for other plant reported values of total anthocyanins in Jabuticaba
product extracts (Rufino et al., 2010a). The flavonoid ranging from 3.67 to 6.16 mg g)1 dry weight. This
composition of several fruits has been reported, but variation may be due to differences in fruit varieties,
more data are needed. Flavonoid-rich plants may be a fruit ripeness, climates and extraction methods. Antho-
good source of antioxidants, helping to increase the cyanins are present in nearly all plant families and,
overall antioxidant capacity of an organism and pro- therefore, in many edible plants. In foods, the main
tecting the organism against lipid peroxidation (Sahreen sources of anthocyanins are berries, such as blackber-
et al., 2010). Feijoa had the highest amount of flavo- ries, grapes and blueberries, and some vegetables, such
noids among the fruits studied, and Grumixama had the as eggplants (aubergine) and avocados. Other anthocy-
lowest value of flavonoids. The one-way anova test anin sources include oranges, elderberries, olives, red
showed differences among the flavonoid contents in the onions, figs, sweet potatoes, mangoes and purple corn
samples. A statistically significant difference (P £ 0.05) (Hendry & Houghton, 1996; Lauro & Francis, 2000).
in flavonoid content among Uvaia, Araçá and Gabiroba The anthocyanin contents in fruits have a wide variation
and between Jabuticaba and Cambuci was not found as follows: raspberry, 10–60 mg 100 g)1 fw; strawberry,
(Table 4). According to Hoffman-Ribani et al. (2009), 15–35 mg 100 g)1 fw; red grape, 30–375 mg 100 g)1 fw;
the main flavonoids found in fresh and processed blueberry, 25–497 mg 100 g)1 fw; avocado, 750
Brazilian fruits are myrecetin, quercetin and kaempfer- mg 100 g)1 fw; and chokeberry, 200–1000 mg 100 g)1
ol, with quercetin being the most common flavonoid fw (Delgado-Vargas & Paredes-López, 2003).
found. Individual anthocyanins have a diverse impact
on the colour and colour stability of many fresh and
DPPH• (free radical scavenging) assay
processed fruits and vegetables, and they have potential
with respect to health benefits (Andersen & Fossen, 1,1-Diphenyl-2-picrylhydrazyl radical is a free radical
2002). Anthocyanins represent a group of widespread that is stable at room temperature and that produces a
natural PC in plants. Quantitative differences of antho- violet solution in ethanol. DPPH• is reduced in the
cyanin concentrations were observed among the fruits presence of an antioxidant molecule giving rise to
(P £ 0.05). Uvaia had the lowest anthocyanin content uncoloured ethanol solutions. The use of DPPH•
(4.77 ± 0.41 mg 100 g)1 fw), and the highest anthocy- provides an easy and rapid way to evaluate antioxidant
anin content was found in Jabuticaba (342.23 ± 2.31 activity (Silva et al., 2005). The radical scavenging
mg 100 g)1 fw). There has been little information activity of ethanolic fruit extracts against the DPPH•
reported about anthocyanins in Brazilian fruits. The radical is shown in Figure 2. All fruit extracts had free
Figure 2 Antioxidant activity and EC50 val-

ues of the fruits based on the DPPH• assay
(ND represents not determined). Bars with
different letters are significantly different
(P £ 0.05).
radical scavenging activity, and a good linear fit of the

b-Carotene-linoleic acid assay
data was observed (R2 ranged from 0.966 to 0.998). The
antioxidant activity varied from 30.17% (BHT) to The b-carotene bleaching method is widely used in
96.68% (Trolox). The best antiradical efficiency values laboratories around the world. Because high tempera-
were found in Jabuticaba and Uvaia fruit extracts with tures are not required, the antioxidant capacity of
an antioxidant activity of 72.41% and 71.08%, respec- thermo-sensitive vegetable extracts may be determined
tively. A statistical difference between the antioxidant and qualitatively evaluated (Hassimotto et al., 2005). In
activity of Jabuticaba and Uvaia extracts was not found this reaction, the oxidation of linoleic acid generates
(P > 0.05). When compared to the fruits analysed, the peroxyl-free radicals because of the abstraction of the
controls (Trolox and propyl gallate) had higher antiox- hydrogen atom from linoleic acid diallylic methylene
idant activity with the exception of BHT. Importantly, a groups. The free radical then oxidises the highly
correlation between PC content and free radical scav- unsaturated b-carotene. The presence of antioxidants
enging activity was not found in this study (r = 0.36). in the extract minimises the oxidation of b-carotene by
Different correlations between PC and DPPH• activity hydroperoxides. Hydroperoxides formed in this system
have been reported by Ranilla et al. (2010) when are neutralised by the antioxidants from the extracts
studying medicinal plants, herbs and spices. They (Kumaran & Karunakaran, 2006). In this study, anti-
obtained a correlation in medicinal plants and spices oxidant capacity was determined from the ability of
(r = 0.81 and 0.86, respectively), but they did not report samples to inhibit b-carotene bleaching caused by free
the same behaviour for herbal teas (P > 0.05). Liu et al. radicals generated during linoleic acid peroxidation.
(2007) studied the total phenolic content and DPPH• Figure 3 shows the antioxidant activity of the fruit
radical scavenging activity of lettuce, and they reported extracts as measured by the bleaching of the b-carotene-
that the total PC content was not highly correlated with linoleic system. The absorbance rapidly decreased in
the DPPH• scavenging ability of lettuce (R2 = 0.27) samples without an antioxidant, and the colour was
when they compared their data in a linear correlation retained for a long time in the presence of an antiox-
model. A negative and significant correlation between idant. Among the positive controls used in this test,
polyphenols and DPPH• scavenging ability was BHT had the highest value of antioxidant activity
observed by Rufino et al. (2010a) in an investigation (inhibition of the free radical peroxidation of lipids):
of eighteen non-traditional tropical fruits from Brazil. 94.94 ± 0.91% at 100 ppm. Lipid hydroperoxides are
Because all of the methods for the assessment of PC and non-radical intermediates derived from unsaturated
antioxidant capacity are based on redox properties, fatty acids, phospholipids, glycolipids, cholesterol esters
there should be some correlation between the PC and cholesterol. Six fruit extracts had high antioxidant
content (free or total) and antioxidant capacity as activity (greater than 80%). Jabuticaba and Uvaia
measured by the individual methods (Stratil et al., showed 97.51 ± 0.61% and 94.71 ± 2.15% inhibition
2007). However, data in the literature with regard to of lipid peroxidation, respectively. The same behaviour
the relation between the concentration of PC and was observed for both fruits in the DPPH• test. The
antioxidant activity are contradictory. While some antioxidant activity displayed by fruit extracts or other
authors have observed a high correlation between PC antioxidants depends on several factors, such as con-
concentration and antioxidant activity, others have centration, temperature, hydrophobic character, hydro-
found no direct correlation or only a weak correlation philic character or amphipathic character, in addition to
(Stratil et al., 2007). Based on the concentration at the presence of synergists and chemical nature of the
which 50% radical scavenging occurred, a relation food or medium to which the synergists are added
between the antioxidant activity and EC50 values was (Lagouri & Boskou, 1995).
clear. The antioxidant activity values were inversely
proportional to the EC50 values. The EC50 values ranged
Antibacterial activity
from 45.49 ± 1.34 lg mL)1 (Uvaia extract) to
121.24 ± 3.12 lg mL)1 (Gabiroba extract). The fruit Diffusion methods are extensively used to investigate the
extracts had similar antioxidant properties, except for antibacterial activity of natural substances and plant
Uvaia and Jabuticaba. A statistical difference in the extracts. These assays are based on the use of discs or
EC50 values of Jabuticaba and Uvaia was not found holes as reservoirs containing the solutions of substances
(P > 0.05). The Jabuticaba and Uvaia fruit extracts had to be examined. According to the antibacterial tests,
good antioxidant activity in the DPPH• system and low K. pneumoniae was the most sensitive bacterium to the
EC50 values. These values were lower than those fruit extracts. Among the seven fruits studied, only the
reported by Prado (2009) when studying the phenolic Jabuticaba extract showed a slight inhibitory effect when
compound content and antioxidant activity of pineap- compared to the positive and negative controls. Never-
ple, acerola, mango, passion fruit, melon, guava and theless, no antibacterial activity was observed in the
pitanga fruit extracts. tests against E. coli and S. aureus. Interestingly, the
Figure 3 Antioxidant activity of coupled oxi-

dation of b-carotene and linoleic acid. Bars
with different letters are significantly different
(P £ 0.05).
concentration of PC extract used in this study was not as these and other unexplored fruits with antioxidant and
high as the concentrations used in other studies. The antimicrobial activities is of particular interest for future
results from this study were similar to results reported studies.
by Rauha et al. (2000), who screened antimicrobial
effects of Finnish plant extracts and reported that the
Acknowledgements
majority of the plant extracts showed no or slight
antibacterial activity against E. coli and S. aureus. In The authors would like to thank the National Council
addition, all of the samples in their study were extracted for Scientific and Technological Development (CNPq;
from food plants, and only a few samples had clear Process Number 501535 ⁄ 2009-8) and Araucaria Foun-
antimicrobial activity. dation for financial support. We would also like to
thank Sitio do Belo for providing the fruits for this work
and Dr Lı́via Bracht for her assistance in the microbi-
Conclusion
ological analysis.
The results of this study show a wide range of total
phenolic content, flavonoid content, anthocyanin con-
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International Journal of Food Science and Technology 2011, 46, 1529–1537 1529

These fruits represent an opportunity for local produc-

Plant material Phenolic compound quantification

Figure 1 Volume vs. time of extraction of

Table 4 Total phenolics, ﬂavonoids and

Uvaia 373.40 ± 14.10e 24.09 ± 0.91c 58.72 ± 2.67b 4.77 ± 0.41e

Figure 2 Antioxidant activity and EC50 val-

radical scavenging activity, and a good linear ﬁt of the

Figure 3 Antioxidant activity of coupled oxi-

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