Food and Chemical Toxicology 50 (2012) 2275–2281

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Food and Chemical Toxicology

journal homepage: www.elsevier.com/locate/foodchemtox

Antigenotoxic effect of acute, subacute and chronic treatments with Amazonian

camu–camu (Myrciaria dubia) juice on mice blood cells
Francisco Carlos da Silva a, Andrelisse Arruda b, Alexandre Ledel a, Cíntia Dauth a, Nathalia Faria Romão a,
Rafaele Nazário Viana a, Alexandre de Barros Falcão Ferraz a, Jaqueline Nascimento Picada a,
Patrícia Pereira c,⇑
a
Programa de Pós-Graduação em Genética e Toxicologia Aplicada, Universidade Luterana do Brasil (ULBRA), Canoas, RS, Brazil
b
Departamento de Ciências Biológicas, CEULJI/ULBRA, Ji-Paraná, RO, Brazil
c
Departamento de Farmacologia, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil

a r t i c l e i n f o a b s t r a c t

Article history: Myrciaria dubia, a plant native to the Amazon region, stands out as a fruit rich in vitamin C and other
Received 22 January 2012 metabolites with nutritional potential. We evaluated the antioxidant, genotoxic and antigenotoxic poten-
Accepted 11 April 2012 tial of M. dubia juice on blood cells of mice after acute, subacute and chronic treatments. Flavonoids and
Available online 21 April 2012
vitamin C present in the fruit of M. dubia were quantified. In vitro antioxidant activity was evaluated by
DPPH assay. Blood samples were collected for analysis after treatment, and the alkaline comet assay was
Keywords: used to analyze the genotoxic and antigenotoxic activity (ex vivo analysis using H2O2). The amount of
Myrciaria dubia
vitamin C per 100 mL of M. dubia was 52.5 mg. DPPH assay showed an antioxidant potential of the fruit.
Vitamin C
Genotoxicity
No M. dubia concentration tested exerted any genotoxic effect on mice blood cells. In the ex vivo test, the
Mice juice demonstrated antigenotoxic effect, and acute treatment produced the most significant results. After
Comet assay the treatments, there was no evidence of toxicity or death. In conclusion, our data show that M. dubia
juice has antigenotoxic and antioxidant activities, though with no genotoxicity for blood cells. Neverthe-
less, more in-depth studies should be conducted to assess the safety of this fruit for human consumption.
Ó 2012 Elsevier Ltd. All rights reserved.

1. Introduction Anthocyanins present during the ripening process alter the

The Amazon region is home to an enormous diversity of fruit
very rich in calories, vitamins, and minerals that afford better qual-
ity of life. However, there is very little knowledge about chemical,
biological and toxicological properties of many of these species.
Myrciaria dubia, is a native Amazonian bush from the Myrtaceae
family commonly known as camu–camu. This species has great
nutritional value, mainly due to the higher levels of potassium
and the ascorbic acid (vitamin C) present in its pulp (Yazawa
et al., 2011; Yuyama et al., 2002). Epidemiological data show that
vitamin C could play a protective role in the development of tu-
mors in humans (Lupulescu, 1993; Duthie et al., 1996). However,
its use should be evaluated specifically in each case as a result of
the many organic and inorganic components in cells that could
model the antioxidant activity of the vitamin, causing it to become
a pro-oxidant. In a clinical trial with camu–camu juice adminis-
tered for seven days, improvements were observed and these ef-
fects were related to the function of high vitamin C levels and of
other compounds in the extract (Inoue et al., 2008).
⇑ Corresponding author. Tel./fax: +55 5133083121.
E-mail address: patipere@yahoo.com.br (P. Pereira).
color of M. dubia, from green to red or purple (Zanatta et al.,
2005). Red–purple fruit are important dietary sources of phenolic
compounds, such as flavonoids and hydroxycinnamic acid deriva-
tives (Pinela et al., 2011). Phenolic compounds may act protecting
biological systems through the stimulation of detoxifying enzymes
that can facilitate the elimination of endogenous and exogenous
toxic compounds or reduce the absorption of toxic compounds
due to cytochrome P450 inhibition. Some phenolic compounds
are able to stimulate DNA repair pathways, through transcription
regulation or mRNA stabilization (Batista et al., 2011; Franke
et al., 2005).
M. dubia fruits contain also carotenoids such as trans-lutein, b-
carotene, violaxantin, luteoxantin and other nutritionally impor-
tant constituents, capable of preventing degenerative diseases
and boost the immune system (Zanatta and Mercadante, 2007).
Oral administration of the M. dubia seeds extract suppressed
experimental edema formation in mice (Yazawa et al., 2011).
However, very few toxicological studies on M. dubia fruit have
been published, even though in the Amazon region it is commonly
consumed by the local population (Justin et al., 2000). Conse-
quently, the objective of this work was to evaluate the genotoxic,
antigenotoxic and antioxidant potential of M. dubia (camu–camu)

0278-6915/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.fct.2012.04.021
2276 F.C. da Silva et al. / Food and Chemical Toxicology 50 (2012) 2275–2281
juice in mice, using three different concentrations along with other
toxicological parameters.
2. Material and methods
2.1. Animals
A total of 120 adult CF-1 mice, male and female, 8–10 weeks of age and weigh-
ing 27.02 ± 2.24 g (female) and 43.46 ± 6.70 g (male), were used. Animals were
housed in plastic cages, with ‘‘ad libitum’’ access to water and food, under a 12-h
light/dark cycle, and at a constant temperature of 22 °C ± 1 °C. Each group was com-
posed by 10 mice (five males and five females) and the protocol for these experi-
ments was approved by the Animal Ethics Committee of the Lutheran University
of Brazil (protocol number CEP-ULBRA 2010-007A).
2.2. Plant material
Samples of M. dubia H. B. K. (McVough) (camu–camu) (Myrtaceae) fruit were
collected in wetlands, quarries and along the banks of the Jarú River in the Munic-
ipality of Jarú, in the central region of the state of Rondônia, Cachoeira Bom Jardim.
Aerial parts, flowers and buds were collected for plant identification and botanical
information. Fruit samples were stored at 20 °C upon analyses, and were admin-
istered to mice. The three concentrations of the juice (25%, 50% and 100%) were pre-
pared always before use so that there was no loss in metabolic properties. Juice was
administered as 0.1 mL/10 g injections.
Dried samples of the plant were identified and stored in the Herbário Central da
Universidade Federal do Mato Grosso (UFMT) under number 29173/2010.
2.3. Phytochemical screening
The plant was subjected to qualitative chemical screening for the identification
of the major classes of active chemical constituents (anthraquinones, alkaloids, car-
diac glycosides, coumarins flavonoids, tannins and saponins). The phytochemical
profile of M. dubia juice was determined by these colorimetric reactions followed
by chromatographic analysis to confirm positive results (Rodrigues et al., 2009).
2.4. Determination of total phenols and tannins
2.4.1. Total phenols
The quantification of phenols was measured according to the Folin–Ciocalteu
method as described in British Pharmacopeia (2007). Briefly, 100 mg of each extract
was dissolved in water and diluted in a 50-mL volumetric flask. The samples were
then filtered through Whatman 9 cm filter paper. After, 5.0 mL of this filtrate were
added to 25 mL with water. Then 2.0 mL of this solution were mixed with 1.0 mL of
Folin–Ciocalteu reagent and 10.0 mL of water and diluted to 25.0 mL with a 15% so-
dium carbonate solution. After 30 min, the absorbance was measured at 760 nm.
The determinations were performed in triplicate (n = 3) and total phenolics content
was expressed as percentage of pyrogallol equivalent (PE).
2.4.2. Total tannins
The quantification of tannins was measured using the Folin–Ciocalteu method
as described in British Pharmacopeia (2007). Then, 0.10 g of casein was added to
the filtrate and shaken vigorously for 60 min. The samples were then filtered
through Whatman 9 cm filter paper and centrifuged for 8 min at 30000 rpm. Five
mL of the filtrate were diluted to 25.0 mL with water. Then, 2.0 mL of this solution
were mixed with 1.0 mL of Folin–Ciocalteu reagent and 10.0 mL of water and di-
luted to 25.0 mL with a 15% sodium carbonate solution. After 30 min, absorbance
at 760 nm was measured. The quantity of tannins corresponded to the difference
between the value found in this analysis and that obtained by measuring total phe-
nols. The determinations were performed in triplicate (n = 3) and the total tannins
was expressed as percentage of pyrogallol equivalent (PE).
2.5. Determination of total flavonoid content
The total flavonoid content was determined according to the colorimetric meth-
od using aluminum chloride (Woisky and Salatino, 1998). In short, a number of
dilutions of quercetin were obtained to prepare a calibration curve. For the determi-
nation of flavonoid content, 50 mg of each sample were dissolved in water and di-
luted with ethanol in 25 mL volumetric flask. Then, 1 mL of aluminum chloride
solution 2.5% was added to 2.0 mL of the stock solution, in a 25 mL volumetric flask.
The final volume was adjusted with ethanol. After 30 min, readings at 425 nm were
performed in a Shimadzu spectrophotometer, for each solution. The determinations
were performed in triplicate (n = 3) and the total contents of flavonoid in the ex-
tracts were expressed as quercetin equivalents (QE) in mg/g of extract.
2.6. Determination of vitamin C
The dosage of vitamin C (ascorbic acid) was performed in accordance with the
methodology described in Farmacopéia Brasileira (2010), using a fruit sample of
0.2 g and dissolving it in a mixture of 100 mL of water free of carbon dioxide and
25 mL of sulfuric acid 10% (w/v). Then, 3 mL of starch SI were added and immedi-
ately titrated with iodine 0.05 M (Sv). Each mL of iodine 0.05 M (sV) was equal to
8.806 mg of C6H8O6.
2.7. Antioxidant assay in vitro
2.7.1. Assay using 2,2-diphenyl-1-picrylhydrazyl (DPPH)
The antioxidant assay of plant material was determined utilizing the DPPH as-
say as described by Rodrigues et al. (2009). From the fruit extract dissolutions mea-
sured at 20,000 lg/mL, a total of 21 different dilutions were prepared in test tubes
in triplicate varying from 100 to 20,000 lg/mL. In a dark environment, 0.1 mL of
each extract dilution was transferred to test tubes with 3.9 mL of the DPPH radical
(DPPH solution of 0.05 mM) and was then homogenized in a test tube shaker. For
the calculation of DPPH we took into account the dilution of the fruit extract –
0.1 mL of each dilution of extract to 3.9 mL of DPPH solution – and utilized the rela-
tion Cinitial Vinitial = Cfinal Vfinal. Therefore, the extract dilution tested ranged between
2.5 and 275 lg/mL. Glass cuvettes were prepared with samples that were moni-
tored until absorbance had stabilized, and readings of these solutions were subse-
quently made utilizing the BIOSPECTRO spectrophotometer, model SP 220, at
515 gm. Using the obtained absorbance values from the different fruit extract dilu-
tions, a linear model was calculated according to the measured concentration of the
total antioxidant activity (TAA) of the M. dubia extracts. The final result was ex-
pressed in grams of fruit (edible portions)/gram of DPPH. The data analysis was car-
ried out by regression (line equation). The larger the consumption of DPPH per
sample, the smaller its EC50 and the larger its antioxidant activity (Sousa et al.,
2007).
2.8. Acute toxicity test
Forty mice were divided into four groups of 10 animals each (five males and five
females) for the acute treatment. They received single doses of M. dubia juice (25%,
50% and 100%) or water (control group) by gavage. The general behavior of the mice
was monitored for 1 h after dosing and periodically for 48 h (Regner et al., 2011).
2.9. Subacute toxicity test
Forty mice were divided into 4 groups of 10 animals each (five males and five
females) for the subacute treatment. They were kept under the same conditions
as described previously. The first group was given water and used as control. The
three remaining groups were orally administered (by gavage) M. dubia juice (25%,
50% and 100%) daily for 28 days.
2.10. Chronic toxicity test
Forty mice were divided into 4 groups of 10 animals each (five males and five
females) for the chronic treatment. They were kept under the same conditions as
described previously and the groups were divided as before. Animals received the
oral administrations (gavage) daily for 56 days.
After all treatments, animals were weighted and observed daily for physiologi-
cal and behavioral changes (Regner et al., 2011). The parameters observed were: tail
elevation, body position, equilibrium, tremors, convulsions and piloerection. At the
end of the treatments, mice were sacrificed for blood collection.
2.11. Comet assay
After the acute, subacute and chronic treatments, the animals were sacrificed by
decapitation and samples of blood were collected to evaluate for genotoxic and
antigenotoxic activity using the comet assay described in Tice et al. (2000). The
blood samples (aprox. 100 lL) were placed in 15 lL anticoagulant (heparin sodium
25.000 IU-LiquemineÒ) and 5.0 lL were embedded in 95 lL 0.75% low melting
point agarose (Gibco BRL). The mixture (cell/agarose) was spread on a fully frosted
microscope slide coated with a 300-lL layer of normal melting agarose (1%) (Gibco
BRL). After solidification, slides were transferred to either PBS or 0.25 mM freshly
prepared hydrogen peroxide (H2O2) solution (ex vivo treatment) for 5 min, at 4 °C
as described by Rodrigues et al. (2009). Slides were washed three times with PBS
and then placed in lysis buffer (2.5 M NaCl, 100 mM EDTA and 10 mM Tris, freshly
added 1% Triton X-100 (Sigma) and 10% DMSO, pH 10.0) for 48 h at 4 °C. Subse-
quently, the slides were incubated in freshly made alkaline buffer (300 mM NaOH
and 1 mM EDTA, pH >13) for 20 min, at 4 °C. An electric current of 300 mA and
25 V (0.90 V/cm) was applied for 15 min to induce DNA electrophoresis. The slides
were then neutralized (0.4 M Tris, pH 7.5), stained with silver, and analyzed using a
microscope. Images of 100 randomly selected cells from each animal (50 cells from
each of two replicate slides) were analyzed. Cells were also scored visually accord-
ing to tail size into five classes, ranging from undamaged (0), to maximally damaged
 F.C. da Silva et al. / Food and Chemical Toxicology 50 (2012) 2275–2281
(4), resulting in a single DNA damage score for each animal, and consequently each
group studied. Therefore, the damage index (DI) can range from 0 (completely
undamaged, 100 cells  0) to 400 (with maximum damage, 100  4). The damage
frequency (DF) was calculated based on number of cells with tail versus those with
no tails (Rodrigues et al., 2009).
2.12. Statistical analysis
Results are expressed as means ± S.D and statistical significance was deter-
mined by a One-Way Analysis of Variance (ANOVA) complemented by the Dun-
nett’s multiple comparisons test. In all comparisons, p < 0.05 was considered as
indicating statistical significance.
3. Results
3.1. Phytochemical analysis
The phytochemical analyses of the M. dubia fruit indicated a
presence of flavonoids and saponins. The total phenolics and
tannins contents were 10.30 ± 1.09 and 1.49 ± 0.24 mg of PE%.
The amount of total flavonoids was 0.57 ± 0.02 mg of QE /g extract.
Also, M. dubia juice in natura was shown to have 52.5 mg of vitamin
C in 100 mL of juice.
3.2. DPPH antioxidant assay in vitro
The results of the DPPH free radical assay are presented in Fig. 1.
The results demonstrated that M. dubia juice has a dose-dependent
scavenger activity in relation to DPPH free radical. The total antiox-
idant activity of M. dubia was 9.72 g of fruit/g of DPPH, with EC50
being 116.71 lg/mL.
3.3. Acute toxicity test
After acute treatment there was no evidence of toxicity and no
animal died.
3.4. Subacute toxicity test
No male or female mice treated with M. dubia juice died during
the 28 days of observation. The animals did not show any changes
in their general appearance during the observation period. Con-
cerning the control group, an increase in weight for male mice that
were treated with the 100% juice concentration (p < 0.01) was de-
tected. For the other groups, no significant change was observed.
3.5. Chronic toxicity test
After 56 days of observation, no male or female animals treated
with M. dubia juice died. The animals did not show any changes in
their general appearance during the observation period. No signif-
Fig. 1. Antioxidant activity of M. dubia juice using the DPPH method. Absorbance
value at 515 gm as a function of the concentration of M. dubia juice in mg/L.
2277
icant weight gain was reported for any of the treated groups, in
relation to the control group.
3.6. Comet assay
Fig. 2 shows the comet assay (damage index) results for the
acute, subacute and chronic oral administration of M. dubia juice
in mice. None of the juice concentrations produced genotoxic effect
after the three period treatments. In the analysis for antigenotoxic
potential, the results demonstrated a significant decline in damage
index (Fig. 2A1 and A2) for all of the groups treated with the juice
after the acute treatment in relation to the control group water +
H2O2. Considering the subacute treatment, there was a significant
decrease in damage index only in both male and female mice trea-
ted with the concentration of 50%, in comparison to water + H2O2
(Fig. 2B1 and B2). Chronically administered juice concentrations
of 50% and 100% reduced the genotoxicity induced by H2O2
(Fig. 2C1 and C2). Although damage caused by juice 25% was not
significant, lower DI were observed after both subacute and
chronic treatments.
Fig. 3 shows the values of damage frequency after acute, suba-
cute or chronic treatments of M. dubia juice in mice. Since M. dubia
juice did not cause a genotoxic effect, there was no increase in the
damage frequency to the DNA detected after the acute, subacute
and chronic treatments. In the test for antigenotoxic potential,
the results demonstrated a significant decrease in damage
frequency (Fig. 3A1 and A2) in groups treated with the juice in rela-
tion to the control group water + H2O2, after acute treatment. After
subacute treatment, only the groups treated with the juice concen-
tration of 50% presented a significant decrease in the damage fre-
quency (Fig. 3B1 and B2). After chronic treatment, a reduction in
damage frequency was observed in both male and female mice
that received the juice concentration of 100% (Fig. 3C1 and C2);
there was no significant difference between male and female, in
both the analysis for genotoxic and antigenotoxic activity.
4. Discussion
Most known organic substances have been produced from nat-
ural sources, and the plant kingdom has become a major contribu-
tor to the provision of bioactive molecules whose synthesis might
be too complex or impractical in laboratory. These products found
in nature reveal an almost unbelievable range of diversity in terms
of structure and biological properties that result in their great
importance (Heitzman et al., 2005).
Various antioxidant compounds are natural components of
plants in general, particularly fruit (Maurya and Devasagayam,
2010; Nunes et al., 2011). Nutritional guidelines recommend the
intense consumption of fruits and vegetables so as to prevent dis-
ease and maintain a healthy condition (Arachi et al., 2010). Consid-
ering the high vitamin C content in M. dubia, and the capacity of
the substances in its chemical composition to eliminate free radi-
cals, we decided to use the DPPH assay to analyze this possibility
in our research. The in vitro DPPH test revealed a dose-dependent
activity of M. dubia juice against the radical DPPH, confirming its
antioxidant potential in natura. This result can be attributed to
vitamin C, flavonoids and phenolic compounds present in this fruit
species (Genovese et al., 2008; Chirinos et al., 2010; Gonçalves
et al., 2010). Many biological properties, such as antioxidant,
anti-inflammatory and hypocholesterolemic activities, have al-
ready been attributed to the secondary plant metabolites. The anti-
oxidant activity of flavonoids is a result of their ability to scavenge
free radicals, to act as hydrogen donors, and to chelate metals,
which reduces the potential for the occurrence of chronic diseases
(Alonso and Gonzalo, 2002; Rice-Evans et al., 1996).
2278 F.C. da Silva et al. / Food and Chemical Toxicology 50 (2012) 2275–2281

Fig. 2. Genotoxic/antigenotoxic effect of M. dubia juice after (A) acute, (B) sub-acute, and (C) chronic treatments as evaluated by the comet assay in male and female mice.
Damage index can range from 0 (completely undamaged, 100 cells  0) to 400 (with maximum damaged 100  4). Statistically significant difference compared to control
group ⁄p < 0.05, ⁄⁄p < 0.01, ⁄⁄⁄p < 0.001. (One-Way ANOVA followed by Dunnett’s test).

M. dubia belongs to the Myrtaceae family, and the fruit has great
nutritional potential, easily adapted and included in human diets.
However, since it is a wild fruit, investment in research in order
to better evaluate its nutritional value in relation to its antioxidant
and nutritional content would be relevant, especially since it is al-
ready being consumed in regions of the Amazon Rainforest. For
this reason, in this work we also investigated the toxicological
parameters of acute, subacute and chronic treatment of the M. du-
bia juice (25%, 50%, and 100%) in male and female mice.
The genotoxic and antigenotoxic action of M. dubia juice on
mice blood cells was analyzed using the comet assay. No concen-
tration tested demonstrated genotoxic action in blood cells (Figs.
2 and 3). In the present study, the vitamin C concentration found
in M. dubia juice was 52.5 mg/10 mL of (corresponding to 0.5 mg/
 F.C. da Silva et al. / Food and Chemical Toxicology 50 (2012) 2275–2281 2279

Fig. 3. Genotoxic/antigenotoxic effect of M. dubia juice after (A) acute, (B) sub-acute, and (C) chronic treatments as evaluated by the comet assay in male and female mice.
Damage frequency was calculated based on the number of cells with tail vs those with no tail. Statistically significant difference compared to control group ⁄p < 0.05,
⁄⁄
p < 0.01, ⁄⁄⁄p < 0.001. (One-Way ANOVA followed by Dunnett’s).

mL). This amount can be considered the highest level of this com-
pound in fruit. It is possible that the lack of damage to the DNA of
blood cells can be credited to interactions with other metabolic
compounds present in M. dubia juice, promoting a reduction in
the pro-oxidant action of vitamin C (Aranha et al., 2010). Moreover,
several studies showed the efficacy of vitamin C in reducing geno-
toxicity. For instance, a protective effect of vitamin C was observed
against ethyl methane sulphonate-induced genotoxicity in fish
(Guha and Khuda-Bukhsh, 2002).
The ex vivo test with M. dubia juice demonstrated an antigeno-
toxic effect after acute treatment in all of the concentrations. For
the subacute and chronic treatments, only the concentrations of
2280 F.C. da Silva et al. / Food and Chemical Toxicology 50 (2012) 2275–2281
50% and 100% played a modulator role in the genotoxicity induced
by the H2O2, although juice 25% exhibited a slight antigenotoxic ef-
fect. These results suggest the protective capacity of M. dubia juice
against genetic damage in the acute treatment or with repeated
doses over time. Other fruits rich in vitamin C, like acerola (Malpig-
hia glaba L.), showed a protective effect against DNA damage
caused by H2O2 (Nunes et al., 2011), and similar tests that assesses
the effect of orange juice on blood cells of rats proved that it re-
duces the DNA damage induced by the mutagenic agents methyl
methanesulfonate and cyclophosphamide (Franke et al., 2005). M.
dubia also demonstrated antigenotoxic activity.
The phytochemical analysis revealed the presence of saponins,
flavonoids and tannins in M. dubia juice. These compounds present
in its fruits, together with vitamin C, probably had a fundamental
role in the reduction of DNA damage induced by H2O2, which
was demonstrated by the comet test in three treatments. As result,
it was possible to attribute an antioxidant potential to this fruit.
In the acute treatment, the antigenotoxic effect can be associ-
ated to the elevated levels of vitamin C as well as the flavonoids
and phenolic compounds present in this fruit that are able to re-
move free radicals. This effect can be related to antioxidant mech-
anisms of the phenolic compounds: chelation of metal and/or the
elimination of free radicals, both of which can decrease oxygen
toxicity for the cells (Khokhar et al., 2003; Maurya and Devasaga-
yam, 2010). Concerning subacute and chronic treatments, which
are more prolonged, it is possible that the protective effect is a re-
sult of the induction of antioxidant enzymes, triggered by the
anthocyanin flavonoids present in this fruit. These compounds
may add protective effect, besides having endogenous scavenger
components, and also interfere in the free radicals production sys-
tems, increasing the performance of endogenous antioxidants
(Volp et al., 2008). Protective effects to the DNA by complex mix-
tures like fruit juices have been attributed to the combination of
carotenoids, phenolic compounds and vitamins (Halliwell, 2001).
The high values of vitamin C present in M. dubia fruit may play a
role in hydroxylation and oxidation–reduction reactions by donat-
ing electrons in intra-and extracellular reactions. Vitamin C can
also act to protect other antioxidants such as vitamin A, vitamin
E and fatty acids from free radicals, including superoxide, hydrogen
peroxide, hydroxyl radical, peroxyl radical and singlet oxygen (Sies,
1997; Halliwell, 2001). The antioxidant potential of a considerable
number of other fruit species, like acerola (Malpighia glabra L.), has
been associated to high level of vitamin C and other important
compounds, like flavonoids (Nunes et al., 2011).
Earlier studies with healthy individuals and smokers demon-
strated that M. dubia juice can have stronger antioxidant and
anti-inflammatory properties in comparison to equivalent vitamin
C pills. These effects can be attributed to the unknown existence of
antioxidant substances in addition to vitamin C (Inoue et al., 2008).
The Amazon region is home to a large variety of fruit species,
with diverse smells and flavors, which represent potentially
interesting economic alternatives and that may be seen as rele-
vant aspects in terms of raising awareness as to the importance
of the region. Several species of fruit consumed in the region
have been shown to exert significant biological action and, there-
fore, they may be an important source in natural medicine pro-
duction processes. Here, the investigation of genotoxic/
antigenotoxic effect of the dubia juice was based on the way
the general population consumes this fruit. Considering that M.
dubia juice contains 52.5 mg of vitamin C in every 100 mL of
juice, it would be of great importance in the provision of this
micronutrient as well as other nutrients present in the fruit, if
supplemented daily in human diets. However, more studies aim-
ing to investigate the safe consumption of this species by the hu-
man population are necessary.
Conflict of Interest
The authors declare that there are no conflicts of interest.
Acknowledgements
This work was supported by CNPq (Conselho Nacional de
Desenvolvimento Científico e Tecnológico), and FAPERGS (Fun-
dação de Amparo à Pesquisa do Estado do Rio Grande do Sul);
Brazil.
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