ArticlePDF Available

Toxicity of the goji berry fruit associated with artificial excipients and dried without additives



The aim of this study was to evaluate the cytotoxic and genotoxic potential of goji berry fruit-based pharmaceutical powders obtained from three pharmaceutical laboratories. The product A was tested at concentrations of 0.012; 0.025 and 0.05 g mL⁻¹, and B and C at concentrations 0.02; 0.04 and 0.08 g mL⁻¹. It was also evaluated the tea of the dried goji berry fruit (non-additives) in the concentrations 0.035; 0.07 and 0.14 g mL⁻¹ for comparison to the results obtained with powdered goji berry. Tea concentrations in the two exposure times did not cause inhibition of cell division nor cellular alterations to meristem tissues. For the industrialized goji products, all concentrations analyzed caused significant antiproliferative effect to the tissues evaluated at the shortest time of analysis. There were no significant cellular changes in tissues exposed to industrialized goji. Therefore, under the conditions of analysis, goji berry powder, at the three concentrations evaluated, was cytotoxic to root meristems. © 2018, Eduem - Editora da Universidade Estadual de Maringa. All rights reserved.
cta Scientiarum
ISSN on-line: 1807-863X
Doi: 10.4025/actascibiolsci.v40i1.37844 BIOTECHNOLOGY
Acta Scientiarum. Biological Sciences, v. 40, e37844, 2018
Toxicity of the goji berry fruit associated with artificial excipients
and dried without additives
Débora Dayane Araújo de Moura1, Cleidiane Josefa dos Santos Veloso1, Valtânia Ana de
Oliveira1, Eduarda Sousa e Silva1 and Ana Paula Peron1,2*
1Laboratório de Citogenética e Mutagênese, Curso de Ciências Biológicas, Universidade Federal do Piauí, Picos, Piauí, Brazil. 2Programa de Pós-
graduação em Genética e Melhoramento, Centro de Ciências Agrárias. Campus Ministro Petrônio Portella, Universidade Federal do Piauí, 64049-
550, Campus Universitário Ministro Petrônio Portella, Bairro Ininga, Teresina, Piauí, Brazil. *Author for correspondence:
ABSTRACT. The aim of this study was to evaluate the cytotoxic and genotoxic potential of goji berry
fruit-based pharmaceutical powders obtained from three pharmaceutical laboratories. The product A was
tested at concentrations of 0.012; 0.025 and 0.05 g mL-1, and B and C at concentrations 0.02; 0.04 and 0.08
g mL-1. It was also evaluated the tea of the dried goji berry fruit (non-additives) in the concentrations 0.035;
0.07 and 0.14 g mL-1 for comparison to the results obtained with powdered goji berry. Tea concentrations
in the two exposure times did not cause inhibition of cell division nor cellular alterations to meristem
tissues. For the industrialized goji products, all concentrations analyzed caused significant antiproliferative
effect to the tissues evaluated at the shortest time of analysis. There were no significant cellular changes in
tissues exposed to industrialized goji. Therefore, under the conditions of analysis, goji berry powder, at the
three concentrations evaluated, was cytotoxic to root meristems.
Keywords: Lycium barbarum L.; natural pharmaceutical product; cytotoxicity; genotoxicity; meristematic tissue.
Toxicidade do fruto goji berry associado a excipientes artificiais e desidratado sem
RESUMO. Objetivou-se na presente pesquisa avaliar, em células meristemáticas de raízes de A. cepa, nos
tempos de exposição 24 e 48 horas, o potencial citotóxico e genotóxico de produtos farmacêuticos do fruto
goji berry em pó, provenientes de três laboratórios farmacêuticos. O produto A foi avaliado nas
concentrações 0,012; 0,025 e 0,05 g mL-1, e B e C nas concentrações 0,02; 0,04 e 0,08 g mL-1. Avaliou-se
também o chá do fruto seco de goji (não aditivado), nas concentrações 0,035; 0,07 e 0,14 g mL-1, para
comparação com os resultados obtidos do fruto em pó. Verificou-se que o chá nas concentrações avaliadas,
nos dois tempos de exposição estabelecidos, não ocasionaram inibição da divisão celular e nem alterações
celulares aos meristemas de raízes. Para os goji industrializados, todas as concentrações analisadas causaram
efeito antiproliferativo significativo aos tecidos avaliados logo no menor tempo de análise considerado.
Nenhum dos produtos industrializados causou número significativo de alterações aos meristemas
analisados. Assim, os goji em pó foram citotóxicos ao bioensaio utilizado por terem acarretado relevante
instabilidade genética aos meristemas de raízes.
Palavras-chave: Lycium barbarum L.; produto farmacêutico natural; citotoxicidade; genotoxicidade; tecido meristemático.
Lycium barbarum L. popularly known as goji berry
or goji, is a shrub Solanaceae, two to four meters
high, widely cultivated in China and the Himalayas.
Its fruits, also commonly referred to as goji berry
and goji, are exported to all continents as spices and
mainly for medicinal purposes, because they contain
high concentrations of flavonoids and relevant
antioxidant potential. They are also sources of an
analog of ascorbic acid, 2-O-β-D-Glucopyranosyl
acid or AA2βG, as well as, of the carotenoid
zeaxanthin (Nascimento et al., 2016) and
polysaccharides, which are efficient antioxidant,
hypoglycemic and anxiolytic (Amagase &
Farnsworth, 2011; Nascimento et al., 2016).
Moreover, since the beginning of this decade, goji
fruits have been prescribed by medical doctors
and nutritionists, and therefore widely marketed
in drugstores around the world, in the form of
industrialized natural powdered pharmaceutical
products, as a potent supplement in weight
reduction and control (Carnés et al., 2013).
Page 2 of 7 Moura et al.
Acta Scientiarum. Biological Sciences, v. 40, e37844, 2018
As a standard procedure, pharmaceuticals for internal
use, such as goji powder, during industrialization are
added with chemical excipients, which are inactive
micro-ingredients devoid of therapeutic activities and
added intentionally. These ingredients are added for the
purpose of making such products palatable and
protected from the action of undesirable
microorganisms, among other characteristics (Araújo &
Borin, 2012; Vasconcelos, Rolim, & Peixoto, 2012).
Among the excipient additives extensively used by
pharmaceutical laboratories are preservatives, colorants,
flavorings, sweeteners, thickeners, emulsifiers and
stabilizers (Balbani, Stelzer, & Montovani, 2006; Araújo
& Borin, 2012).
However, in their technical regulations, the
Agência Nacional de Vigilância Sanitária (ANVISA) and
the Codex Alimentarius, declare that the flavoring,
sweetening, anti-wetting and acidulant additives,
although released for use, raise a number of doubts
about their potential cytotoxic, genotoxic and
mutagenic effects. It also highlights the relevance
and urgency of conducting research to evaluates the
cytotoxic and genotoxic effects of products added
with artificial microingredients (Brasil, 2007). The
adverse effects observed from toxicity assessments are
very relevant, since they represent important
parameters in the elaboration and/or modification of
documents that regulate the use of excipients by the
industries (Konishi, Hayashi, & Fukushima, 2013;
Bezerra, Malaquias, Sousa, & Peron, 2016; Sales,
Santos, Sousa, Silva, & Peron, 2017).
However, in a broad search in the scientific
literature, we verified that to date there are no
studies evaluating cytotoxicity and genotoxicity of
goji berries marketed powdered in industrialized
form. In contrast, for fresh goji berry fruit, we found
studies that evaluate its toxic potential at the cellular
level, such as the work of Sayeed et al. (2017), who
evaluated the toxicity of this fruit through in vivo
micronucleus test and comet assay using colon cells
from Wistar rats and reported that the aqueous goji
extract was neither cytotoxic nor genotoxic to the
test systems used. In addition, Yang, Zhao, Chen,
Chan, and Wu (2015) and Potterat (2010) verified
that goji fruit tea did not alter cell division and was
antigenotoxic to normal cell lines. Thus, it is
relevant to evaluate, through appropriate toxicity
bioassays, the cytotoxicity and genotoxicity of the
powdered fruit added with excipient compounds, in
order to determine if this has toxic potential
significantly different from its fresh form.
Evaluations like these can contribute to the safe and
effective consumption of these industrialized natural
products by the population.
Plant bioassays are appropriately sensitive and
simple in monitoring the toxic effects at the cellular
level of chemical compounds (Caritá & Marin-
Morales, 2008; Campos-Ventura, & Marin-Morales,
2016). Among them, the root meristem of Allium
cepa L. (onion) is considered in the scientific circle as
an efficient bioassay for the initial screening of the
genetic toxicity of chemical compounds due to the
low number of chromosomes (2n = 16), which
favors the detection of mitotic spindle or aneugenic
defects, and disturbances in the cell proliferation
index (Neves, Ferreira, Lima, & Peron, 2014;
Bianchi, Mantovani, & Marin-Morales, 2015). It is a
test system accepted internationally by research
agencies as an instrument of evaluation with
accurate sensitivity to analyze the cytotoxicity and
genotoxicity of a substance of interest, since the
results obtained often show satisfactory similarity to
those obtained through animal testing systems and
cell cultures (Herrero et al., 2012; Lacerda,
Malaquias, & Peron, 2014; Tabrez et al., 2011;
Bianchi et al., 2015; Campos-Ventura et al., 2016;
Santana et al., 2016). As an example, we can cite the
researches conducted by Gomes, Oliveira, Carvalho,
Menezes, and Peron (2013) and Oliveira, Alves,
Lima, Sousa, and Peron (2013), which evaluated in
root meristem cells of A. cepa the toxic potential of
synthetic colorings used in the food and
pharmaceutical industries and obtained results
similar to those obtained via animal testing systems
and via cell cultures.
In this context, the present study aimed to
evaluate, in root meristem cells of A. cepa, the
cytotoxicity and genotoxicity of natural
pharmaceutical products of goji berry fruit powder
obtained from three different pharmaceutical
laboratories of relevant performance in the Brazilian
and international marketing of medicines. The
evaluation of dried goji berry, without additives, tea
was also performed with the purpose of comparing
the results with data observed for the industrialized
form considering the same test system.
Material and methods
The dried goji fruit, without addition of artificial
excipients, was acquired in an herbal store, in the
city of Teresina, State of Piauí, Brazil, specialized in
the commercialization of natural products. In the
industrialized form, the goji fruit marketed in
powder form, was acquired in a unit of a national
drugstore network in the municipality of Picos,
State of Piauí, Brazil. The industrialized products,
from three pharmaceutical laboratories, were
discriminated in the present research as A, B and C.
Toxicity of the goji berry Page 3 of 7
Acta Scientiarum. Biological Sciences, v. 40, e37844, 2018
For determination of goji concentrations to be
analyzed for toxicity, the form of preparation and
ingestion indicated on the labels of each product,
fresh and industrialized, was used as the definition
parameter. For the preparation of the tea, 70 grams
of the dried fruit should be mixed with a liter of
boiling water. Thus, three concentrations of the tea
were defined for analysis, which were 0.035; 0.07
and 0.14 g mL-1. Regarding the industrialized goji
for laboratory A, the suggested concentration was 70
grams fruit powder for 200 mL water. Thus, the
analysis concentrations for A were 0.012; 0.025 and
0.05 g mL-1. For laboratories B and C, the ideal
concentration suggested for consumption was 5 g of
goji powder for 200 mL water, and concentrations
for toxicity analysis were 0.012; 0.025 and 0.05
g mL-1. To obtain all the concentrations evaluated in
the present study, we used distilled water.
For toxicity analysis, onion bulbs were placed in
aerated bottles with distilled water, at room
temperature (± 27ºC), to obtain 2.0 cm long roots. For
the analysis of each goji sample, an experimental group
with five onion bulbs was established. Before placing
the roots in contact with their respective goji samples
(treatments), some roots were collected and fixed to
serve as control of the bulb itself. Then, the remaining
roots were placed in contact with their respective
treatments for 24 hours, a procedure called 24 hours
exposure time. After 24 hours, some roots were
collected and fixed. After, the remaining roots of each
bulb were returned to their respective treatments,
where they remained for additional 24 hours, which
was called 48 hours exposure time. Subsequently, roots
were again collected and fixed. The 24 and 48 hours
exposure times were chosen with the purpose of
evaluating the action of goji in more than one cell
cycle. Roots were fixed in Carnoy 3: 1 (ethanol: acetic
acid) for 24 hours. In each collection, on average, three
roots were taken per bulb.
The slides, on average 03 per bulb, were mounted
according to Guerra and Souza (2002) and analyzed
under an optical microscope (Zeiss brand) using 400x
objective lens. For each onion bulb, we analyzed 1,000
cells, totaling 5,000 cells for each control, 24 hours
exposure time and 48 hours exposure time of each
treatment group analyzed. Thus, for each goji
concentration, we analyzed a total of 15,000 cells. Cells
were observed in interphase, prophase, metaphase,
anaphase and telophase. From this analysis, the mitotic
index (MI) was determined by means of the following
equation: (total number of cells in mitosis ÷ total
number of cells analyzed) x 100. The MI value was the
parameter used for the determination of the cytotoxic
potential of goji berry in the forms analyzed.
In addition, we examined the genotoxicity of goji
by the frequency of cell alterations or mitotic spindle
defects, including C-metaphases, Multipolar anaphase,
Anaphase and telophase bridges, Gene amplifications,
Cells with adhesion, Nuclear buds and Micronuclei.
For the statistical analysis of data on cytotoxicity and
genotoxicity of the samples, we applied the Chi-square
test (χ2), with <0.05 probability level, statistical
program Bioestat, version 5.3.
Results and discussion
As described in Table 01, the root meristems
exposed to the three concentrations of goji berry tea at
the 24 and 48 hours exposure times, when compared
to the observed cell division for their respective
controls, showed no alteration in their mitotic indices.
There was also no significant difference between the
mitotic indices obtained for the two exposure times of
each tea concentration evaluated. Thus, it can be stated
that the evaluated goji teas, under the conditions of
study established, caused no cytotoxicity to root
meristem cells of A. cepa. In addition, there were no
cellular alterations in plant tissues exposed to such
solution. These results corroborate the data of non-
cytotoxicity and genotoxicity of goji fruit found in the
scientific literature (Yang et al., 2015; Potterat, 2010),
previously mentioned in this work.
However, the three concentrations of goji referring
to PL A (Table 01), at 24 and 48 hours of exposure,
significantly reduced the cell division of the
meristematic tissue when compared to the mitotic
indices obtained for their respective controls.
Moreover, for the concentration 0.035 g mL-1 of PL A,
mitotic indices obtained for the two exposure times
evaluated differed from each other, since in the 48
hours exposure time, the cell division was markedly
lower than that observed for the exposure time of 24
hours. However, for PL A at concentrations of 0.07
and 0.14 g mL-1, inhibition of cell division was
observed shortly at the 24 hours exposure time. For the
three concentrations of goji berry products of PLs B
and C (Table 1), the mitotic indices verified for 24
hours exposure time were considered the same as those
obtained for their respective 48 hours exposure time.
In view of this, based on the data described in Table
1, there is a difference in the action of fresh goji berry
in relation to the industrialized goji as to the toxicity
caused, once the excipients added were cytotoxic to the
meristem tissue evaluated for causing a marked
antiproliferative effect at the lowest concentrations and
in the shortest exposure time. It is also noted that all
goji concentrations considered as ideal for
consumption by the pharmaceutical laboratories by
which they were produced have caused significant
inhibition of cell proliferation, demonstrating relevant
cytotoxic potential.
Page 4 of 7 Moura et al.
Acta Scientiarum. Biological Sciences, v. 40, e37844, 2018
Table 1. Number of cells observed in each phase of the cell cycle of the root meristem of Allium cepa exposed for 24 and 48 hours to goji
berry tea and to industrialized powdered goji berry, of the chemical laboratories A, B and C.
CO 3029 599 479 492 401 1971 39.4a
0.03 g mL-1 24h 3465 433 333 398 371 1535 30.7b
48h 3534 429 341 355 355 1466 29.3b
CO 2952 587 581 479 401 1971 41.0a
0.07 g mL-1 24h 3107 499 494 487 371 1535 37.9b
48h 3141 504 483 475 355 1466 29.3b
CO 2780 603 591 594 432 2220 44.4a
0.13 g mL-1 24h 2899 613 559 532 397 2101 42.0 b
48h 2942 594 507 539 418 2058 41.2 b
CO 2827 689 671 581 232 2173 43.5a
0.035 g mL-1 24h 4644 103 93 77 53 326 6.5 b
48h 4792 82 67 59 00 208 4.2 b
CO 3200 321 351 224 191 1087 21.7 a
0.07g mL-1 24h 4883 19 12 03 03 37 0.7 b
48h 4976 07 09 07 05 28 0.5 b
CO 3209 500 705 362 224 1791 36.0 a
0.14 g mL-1 24h 4965 27 07 01 00 35 0.7 b
48h 4958 39 39 00 00 42 0.8 b
CO 4192 333 258 107 110 808 16.2 a
0.02 g mL-1 24h 4738 71 14 08 99 262 2.3 b
48h 4901 73 12 09 05 99 0.5 b
CO 3200 675 542 349 234 1800 21.7 a
0.04 g mL-1 24h 4883 53 44 17 03 117 0.7 b
48h 4976 17 07 00 00 24 0.5 b
CO 3581 522 439 324 134 1419 28.4 a
0.08 g mL-1 24h 4729 92 84 71 24 271 5.4 b
48h 4875 99 19 07 00 125 2.5 b
CO 2306 701 713 609 671 2694 53.9
0.02 g mL-1 24h 4767 55 76 65 37 233 4.7
48h 4948 32 11 01 08 52 1.0
CO 3208 590 427 427 348 1792 35.9
0.04 g mL-1 24h 4872 94 03 07 07 128 2.6
48h 4950 31 15 01 03 50 1.0
CO 2634 709 821 514 322 2366 47.3
0.08 g mL-1 24h 4948 30 17 00 05 52 1.0
48h 4964 25 07 02 02 36 0.7
TR – Treatment; Conc. – Concentration; ET – Exposure Time; ; TCII – Total number of cells in interphase and undifferentiated cells; P – Prophase; M – Metaphase; A – Anaphase;
T – Telophase; CO – Control; IM – MI – Mitotic Index; TCD – Total number of dividing cells; TAC – Total Cellular Alterations; MI – Mitotic Indices; PL – Pharmaceutical
laboratory. Values followed by different letters within the same treatment are significantly different by χ2 test at 5% probability.
According to Caritá and Marin-Morales (2008),
significant alterations are triggered when there is a
pronounced antiproliferative effect in tissues with
intense proliferation and normal metabolic
performance - such as the meristem of roots used in
the present study - exposed to chemical compounds
with potential to cause genetic instability, significantly
compromising the growth and functioning of the
organs in which they are acting. Furthermore, Gomes
et al. (2013); Bezerra et al. (2016) and Carvalho et al.
(2016) state that the inhibition of cell proliferation
triggered by cytotoxic compounds in tissues of intense
cell proliferation and normal functioning and/or
without cellular alterations - once again mentioning
the root meristems used as bioassays in the present
research - is harmful to the organisms by inhibiting or
limiting the replacement of cells, modifying the
production of proteins and, consequently, resulting in
malfunction of the organ or tissue where it is located.
No significant cellular alterations were
registered in the meristematic cells exposed to the
goji concentrations from PLs A, B and C.
Nevertheless, Sales et al. (2017) explain that
inhibition of division in normal tissues occurs by
the action of agents that affect the integrity of the
nuclear spindle during mitosis promoting
significant chromosomal derangement.
Considering that the principle of the cell cycle is
the formation of identical cells, the production of
cells with alterations in structure and/or
chromosome number make the cellular operation
unfeasible and tend to be eliminated from tissues
with normal performance, a condition that is
suggested to explain the result of a marked
Toxicity of the goji berry Page 5 of 7
Acta Scientiarum. Biological Sciences, v. 40, e37844, 2018
antiproliferative effect against the non-significant
frequency of cellular alterations observed in the
present study.
As aforementioned, natural pharmaceutical
products, such as the industrialized goji berries
evaluated herein, contain artificial excipients in their
formulation. Similarly, a toxicity evaluation of
Ginkgo biloba L. leaves in natura and associated
excipients in different bioassays showed that the
industrialized forms were toxic at the cellular level
(manuscript in press). These results corroborate the
results obtained in the present study, suggesting that
the excipients in the industrialization of natural
products have cytotoxic effects.
Results from studies on cellular toxicity of some
excipient compounds used in the formulation of
pharmaceuticals and food products, according to
Brazil (1999), during industrialization will be
reported. However, it is important to state that, with
the exception of colorings and preservatives, all the
microingredients to be reported were insufficiently
evaluated, according to ANVISA, regarding their
toxic potential at the systemic and cellular levels
(Brasil, 2007; Sales et al., 2017).
For colorings, the only authorized for use in
pharmaceutical products in general are Twilight
Yellow, Tartrazine and Red 40, azo additives because
they contain the azo group, a nitro derivative with
the property of producing aromatic amine and
sulphanilic acid, as well as Ponceau 4R, Erythrosine
and the Bright Blue (Gomes et al., 2013; Sardi et al.,
2010). These six dyes showed potential in altering
the turner-over of the cells during interphase,
expressively inhibiting cell division, and the process
of regenerative hyperplasia, which contributed
significantly to the development of cancers in the
digestive tract of rodents (Sardi et al., 2010).
Aspartame, sodium cyclamate, potassium
acesulfame and sodium saccharin are among the
sweeteners permitted as excipients (Balbani et al.,
2006; Vasconcelos et al., 2012). Using cell lines
Caco-2 (colon cells), HT-29 (colon cells) and HEK-
293 (kidney cells), Van EyK (2015) reported that
these sweeteners were cytotoxic and genotoxic to the
cells studied. Corroborating with the results of these
researchers, Zaineddin et al. (2012), through the
comet assay, found that sodium saccharin and
sodium cyclamate were genotoxic and mutagenic to
rodent colon cells, significantly reducing the cell
division of the analyzed tissue.
The anti-wetting agents used in pharmaceuticals
are calcium phosphate, silicon dioxide, calcium
carbonate and magnesium carbonate (Villanova,
Oréfice & Cunha, 2010). There were no studies in
the scientific literature evaluating cytotoxicity and
genotoxicity for the calcium phosphate and calcium
carbonate. In turn, for silicon dioxide, Rajiv et al.
(2016) argued that this compound had the property
to significantly decrease the cell viability of human
lymphocytes in normal cell culture, as well as to
promote significant cellular alterations,
demonstrating a broad genotoxic potential. For the
magnesium carbonate, Ahamed et al. (2015)
observed that this chemical had the ability to reduce
cellular metabolism in normal human liver cell
culture, showing to be significantly cytotoxic.
As for flavorings, the aroma and flavor
ingredients used in medicines are only those of fruit
(Balbani et al., 2006). Sales et al. (2017) and Moura
et al. (2016) evaluated some of these flavorings and
verified that such additives had the property of
inducing significant damage to the mitotic spindle,
and consequently the cell division of human
peripheral blood cells, and were genotoxic to mouse
blood tissue erythrocytes, significantly inducing the
formation of micronucleated cells in the bone
marrow of treated animals. In relation to the
chemical constituents responsible for retarding the
action of microorganisms, enzymes, as well as
physical agents in pharmaceuticals, include
potassium benzoate, sodium benzoate and
potassium nitrate, preservatives which, according to
Mpountoukas et al. (2010) and Zeguin, Yüzbaşioğlu,
Unal, Yilmaz, & Aksoy (2011), were cytotoxic and
genotoxic to normal human peripheral blood cells.
The toxicity results of the mentioned excipients
validate those obtained in the present study for the
industrialized goji evaluated, as they also caused
genetic instability mainly to the cell cycle of the
bioassays in which they were evaluated. No studies
were found in the literature to evaluate the toxicity
at the cellular level of additives of thickening,
emulsifying and stabilizing action.
The data obtained in the present study with the
meristematic root cells of A. cepa showed that the
industrialized goji berries evaluated had a significant
potential to cause toxicity to the cells tested at all
evaluated concentrations, including those indicated
for use by pharmaceutical laboratories.
These results indicate the need to evaluate
powdered goji pharmaceuticals in animal testing
systems, from treatments with longer exposure times,
to verify and deepen the results obtained here.
It is important to emphasize that the results
obtained here with respect to genetic instability
caused by the action of the industrialized goji are of
great relevance since to date there are no toxicity
studies involving such pharmaceutical products.
Page 6 of 7 Moura et al.
Acta Scientiarum. Biological Sciences, v. 40, e37844, 2018
Ahamed, M., Alhadlay, H. A., Ahmad, J., Siddiqui, M. A.,
Khan, S. T., Mussarat, J… Al-Khedhairy, A. A. (2015).
Comparative cytotoxicity of dolomite nanoparticles in
human larynx HEp2 and liver HepG2 cells. Journal of
Applied Toxicology, 35(6), 640-650. doi: 10.1002/jat. 3097
Amagase, H., & Farnsworth, N. R. A. (2011). Review of
botanical characteristics, phytochemistry, clinical
relevance in efficacy and safety of Lycium barbarum
fruit (Goji). Food Research International, 44(7), 1702-
1717. doi: 10.1016/j.foodres.2011.03.027
Araujo, A. C. F., & Borin, M. F. (2012). Influência de
excipientes farmacêuticos em reações adversas a
medicamentos. Brasilia Médica. Brasilia, 49(4), 267-278.
Balbani, A. P. S., Stelzer, L. B., & Montovani, J. C. (2006).
Excipientes de medicamentos e as informações da
bula. Revista Brasileira de Otorrinolaringologia, 72(1),
400-406. doi: 10.1590/S0034-72992006000300018.
Bezerra, M. D. S., Malaquias, G. O., Sousa, J. M. C., &
Peron, A. P. (2016). Cytotoxic and genotoxic potential
of powdered juices. Ciência e Tecnologia de Alimentos,
36(1), 49-55. doi: 10.1590/1678-457X.0006
Bianchi, J., Mantovani, M. S., & Marin-Morales, M. S.
(2015). Analysis of the genotoxic potential of low
concentrations of Malathion on the Allium cepa cells
and rat hepatoma tissue culture. Journal Environmental
Sciences, 36(1), 102-111. doi: 10.1016/j.jes.2015.03.034
Brasil. Agência Nacional de Vigilância Sanitária [ANVISA].
(2007). Resolucão da diretoria colegiada RDC nº. 5, de 15 de
Janeiro de 2007. Brasília: ANVISA. Retrieved on Dec 3,
2016 from
Brasil. Agência Nacional de Vigilância Sanitária
[ANVISA]. (1999). Resolução n. 17, de 30 de abril de
1999 – ANVISA, 1999. Retrieved on Oct 5, 2016 from
Campos-Ventura, B., & Marin-Morales, M. A. (2016).
Micronuclei and chromosome aberrations derived
from the action of Atrazine herbicide in Allium cepa
meristematic cells. SDRP Journal of Earth Sciences &
Environmental Studies, 1(1), 22-28.
Caritá, R., & Marin-Morales, M. A. (2008). Induction of
chromosome aberrations in the Allium cepa test system
caused by the exposure of seeds to industrial effluents
contaminated with azo dyes. Chemosphere, 72(5), 722-
725. doi: 10.1016/j.chemosphere.2008.03.056
Carnés, J., de Larramendi, C. H., Ferrer, A., Huertas, A. J.,
López-Matas, M. A., Pagán, J. A., ... Peña, M. (2013).
Recently introduced foods as new allergenic sources:
sensitisation to Goji berries (Lycium barbarum). Food
Chemistry, 137(1), 130-135. doi: 10.1016.j.ffodchem.
Carvalho, F. R., Moura, A. G., Rodrigues, G. F., Nunes,
N. M., Lima, D. J., Pessôa, C., ... Peron, A. P. (2016).
Are salty liquid food flavorings in vitro antitumor
substances? Anais da Academia Brasileira de
Ciências, 88(3), 1419-1430. doi: 10.1590/0001-
Donno, D., Beccaro, G. L., Mellano, M. G., Cerutti, A.
K., & Bounous, G. (2015). Goji berry fruit (Lycium
spp.): antioxidant compound fingerprint and
bioactivity evaluation. Journal of Functional Foods, 18(1),
1070-1085. doi: 10.1016/j.jff.2014.05.020
Gomes, K. M. S., Oliveira, M. V. G. A. D., Carvalho, F. R. S.,
Menezes, C. C., & Peron, A. P. (2013). Citotoxicity of
food dyes sunset yellow (E-110), bordeax red (E-123),
and tatrazine yellow (E-102) on Allium cepa L. root
meristematic cells. Ciência e Tecnologia de Alimentos, 33(1),
218-223. doi: 10.1590/S0101-20612013005000 012
Guerra, M., & Souza, M. J. (2002). Como observar os
cromossomos: um guia de técnicas em citogenética vegetal,
animal e humana (304p.). Ribeirão Preto, SP:
Herrero, O., Martín, J. P., Freire, P. F., López, L. C.,
Peropadre, A., & Hanzen, M. J. (2012). Toxicological
evaluation of three contaminant of emerging concern
by use of Allium cepa test. Mutation Research, 743(1), 24-
34. doi: 10.1016/j.mrgentox.2011.12.028
Konishi, Y., Hayashi, S. M., & Fukushima, S. (2013).
Regulatory forum opinion piece*: supporting the need
for international harmonization of safety assessments
for food flavoring substance. Toxicologic Pathology,
42(6), 949-953. doi: 10.1177/0192623313495603
Lacerda, L. P., Malaquias, G., & Peron, A. P. (2014).
Antiproliferative action of aqueous extracts of
Hymenaea stigonocarpa Mart. (Fabaceae) on the cell
cycle of Allium cepa L. Anais da Academia Brasileira de
Ciências, 89(3), 1147-1150. doi: 10.1590/0001-
Mpountoukas, P., Pantazaki, A., Kostareli, E., Christodoulou,
P., Kareli, D., Poliliou, S., & Lialiaris T. (2010).
Cytogenetic evaluation and DNA interaction studies of
the food colorants amaranth, erythrosine and tartrazine.
Food Chemical and Toxicology, 48(10), 2934-2944.
doi: 10.1016/j.fct.2010. 07.030
Nascimento, W. M., Souza, L. M. S., Costa, D. D. A. F.,
Oliveira, R., Monte, S. M., Silva, G. C. E., ... Maia
Filho, A. L. M. (2016) Genotoxicity of Goji Berry
(Lycium barbarum) in vivo Mammalian Cells.
International Journal of Pharmaceutical Science Invention,
5(4), 23-26. doi: 16/j.fct.201
Neves, E. S., Ferreira, P. M. P., Lima, L. H. G. M., &
Peron, A. P. (2014). Action of aqueous extracts of
Phyllanthus niruri L. (Euphorbiaceae) leaves on
meristematic root cells of Allium cepa L. Anais da
Academia Brasileira de Ciências, 86(3), 1131-1137.
doi: 10.1590/0001-3765201420130107
Oliveira, M. V. A., Alves, D. D. L., Lima, L. H. G. M., Sousa,
J. M. C., & Peron, A. P. (2013). Cytotoxicity of
erythrosine (E-127), brilliant blue (E-133) and red 40 (E-
129) food dyes in a plant test system. Acta Scientiarum.
Biological Sciences, 35(4). doi: 10.4025/actascibiol
Potterat, O. (2010). Goji (Lycium barbarum and Lycium
chinense): phytochemistry, pharmacology and safety in
Toxicity of the goji berry Page 7 of 7
Acta Scientiarum. Biological Sciences, v. 40, e37844, 2018
the perspective of traditional uses and recent
popularity. Planta Medica, 76(1), 7-19. doi: 10.1055/s-
Rajiv, S., Jerobin, J., Saranya, V., Nainawat, M., Sharma,
A., Makwana, P., ... Mukherjee, A. (2016).
Comparative cytotoxicity and genotoxicity of cobalt
(II, III) oxide, iron (III) oxide, silicon dioxide, and
aluminum oxide nanoparticles on human lymphocytes
in vitro. Human & Experimental Toxicology, 35(2),
170-183. doi: 10.1177/0960327115579208
Sales, I. M. S., Santos, F. K. S., Sousa, J. M. C., Silva, F. C. C.,
& Peron, A. P. (2017). Acute toxicity of grape, plum and
orange synthetic food flavourings evaluated in vivo test
systems. Food Technology and Biotechnology, 55(1), 131-137.
doi: 10.17113/ftb.55.01. 17.4770
Santana, G. M., Deus, M. S. M., Sousa, J. M. C., Ferreira,
P. M. P., Fernandes, H. B., & Peron, A. P. (2016).
Antimitotic and antimutagenic action of the Hymenaea
stigonocarpa bark on dividing cells. Brazilian Journal of
Biology, 76(2), 520-525. doi: 10.1590/1519-6984.23014
Sardi, M., Haldemann, Y., Nordmann, H., Bottex, B.,
Safford, B., Smith, B., & Jasti, P. R. (2010). Use of
retailer fidelity card schemes in the assessment of food
additive intake: sunset yellow a case study. Food
Additives and Contaminants, 27(11), 1507-1515.
doi: 10.1080/19440049.2010.495728
Sayeed, M. A., Bracci, M., Lazzarini, R., Tomasetti, M.,
Amati, M., Lucarini, G., ... & Santarelli, L. (2017). Use
of potential dietary phytochemicals to target miRNA:
Promising option for breast cancer prevention and
treatment? Journal of Functional Foods, 28(1), 177-193.
doi: 10.1016/j.jff.2016.11.008
Tabrez, S., Shakil, S., Urooj, M., Damanhouri, G. A.,
Abuzenadah, A. M., & Ahmad, M. (2011). Genotoxicity
testing and biomarker studies on surface waters: an
overview of the techniques and their efficacies. Journal of
Environmental Science and Health, Part C, 29(3), 250-275.
doi: 10.1080/10590501.2011. 601849
Van Eyk, A. D. (2015). The effect of five artificial
sweeteners on Caco-2, HT-29 and HEK-293
cells. Drug and Chemical Toxicology, 38(3), 318-327.
doi: 10.3109/01480545.2014.966381
Vasconcelos, P. A. F., Rolim, L. A., & Peixoto, M. S. (2012).
Influência dos excipientes multifuncionais no desempenho
dos fármacos em formas farmacêuticas. Revista Brasileira de
Farmácia, 93(2), 136-145.
Villanova, J. C. O., Oréfice, R. L. & Cunha, A. S. (2010).
Aplicações farmacêuticas de polímeros. Polímeros:
Ciência e Tecnologia, 20(1), 51-64.
Yang, R. F., Zhao, C., Chen, X., Chan, S. N., & Wu, J. Y.
(2015). Chemical properties and bioactivities of Goji
(Lycium barbarum) polysaccharides extracted by
different methods. Journal of Funcional Foods, 17(1),
903-909. doi: 10.1016/j.jff.2015.06.045
Zaineddin, A. K., Buck, K., Vrieling, A., Heinz, L.,
FleschJanys, D., Linseisen, J., & Chang-Claude J.
(2012). The association between dietary lignans,
phytoestrogen-rich foods, and fiber intake and
postmenopausal breast cancer risk: a German case-
control study. Nutrition Cancer, 64(5), 652-665.
doi: 10.1080/01635581.2012.683227
Zeguin, N., Yüzbaşioğlu, D., Unal, F., Yilmaz, S., &
Aksoy, H. (2011). The evaluation of the genotoxicity
of two food preservatives: sodium benzoate and
potassium benzoate. Food Chemical and Toxicology,
49(4), 763-769. doi: 10.1016/j.fct.2010.11.040
Received on June 26, 2017.
Accepted on May 2, 2018.
License information: This is an open-access article distributed under the terms of the
Creative Commons Attribution License, which permits unrestricted use, distribution,
and reproduction in any medium, provided the original work is properly cited.
... Preservatives, dyes, flavors, sweeteners, thickeners, emulsifiers, and stabilizers are commonly used in pharmaceutical laboratories (Balbani et al., 2006). Moura et al. (2018) used the root meristem of Alliym cepa L. (onion) as an efficient bioassay for the initial screening of the genetic toxicity. They found that industrialized goji berries at all concentrations, including those indicated for use by pharmaceutical companies, had significant potential for toxicity. ...
Full-text available
Goji (Lycium L.) fruit has been an important element of traditional Chinese medicine for centuries. In Asian countries, they are used as an essential component of a healthy diet, a source of many nutrients. Due to their health-promoting properties and chemical composition (phenolic acids, flavonoids, proanthocyanidins, coumarins, tannins, carotenoids, anthocyanins), they deserved the term superfruit. In recent years, goji berries have also become very popular in Europe and America. The fruit is used primarily after drying and is available in the form of various supplements. Two species of Lycium barbarum L. and Lycium chinense Mill. are cultivated on a larger scale. These species are closely related to each other. They differ slightly in morphological features. L. chinense leaves are longer and wider than L. barbarum. L. chinense fruits are slightly smaller and more elongated. There are several less specific species and botanical varieties in natural sites in central and western China, such as L. barbarum var. aurantiocarpum, L. chinense var. potaninii, L. ruthenicum, L. truncatum. Not only fruits contain biologically active substances, but also other parts of plants, especially leaves. This review highlights the healing properties of the fruits and leaves of these species. The most valuable and most interesting component of goji berries is the water-soluble bioactive polysaccharide complex LBP (Lycium Barbarum Polysaccharides) playing an important therapeutic role. The LBP complex has a beneficial effect on the functions of the immune system, inhibits the growth of cancer cells, has antioxidant properties, improves the function of the digestive tract, well-being and sleep quality. Due to the presence of LBP, goji fruit extracts have a hypoglycemic effect, lowering the content of lipids in the blood serum. The diversity of their use as food, medicinal and cosmetic agents was shown.
Full-text available
The present study evaluated the acute toxicity of grape, plum and orange synthetic flavorings in root meristem cells of A. cepa at the concentrations 3.5; 7.0 and 14.0 mL/kg and exposure times 24 and 48 h, and in bone marrow erythrocytes of mice treated orally for seven days with the concentrations 0.5; 1.0; 2.0; 5.0 and 10.0 mL/kg flavoring. The results from the plant test showed that grape, plum and orange flavorings, in both exposure times, inhibited cell division and promoted the formation of a significant number of micronuclei and mitotic spindle changes. These alterations were observed in at least one exposure time analyzed, demonstrating a significant cytotoxic, genotoxic and mutagenic activity. In relation to the bioassay in mice, animals treated with 2.0; 5.0 and 10.0 mL/kg flavoring died before the seventh day of treatment. The concentrations 0.5 and 1.0 mL/kg of the three additives were Please note that this is an unedited version of the manuscript that has been accepted for publication. This version will undergo copyediting and typesetting before its final form for publication. We are providing this version as a service to our readers. The published version will differ from this one as a result of linguistic and technical corrections and layout editing. cytotoxic to erythrocytes, and significantly induced the formation of micronucleated cells in the bone marrow of animals treated with the microingredients. Therefore, under the study conditions, the flavoring additives grape, plum and orange promoted significant toxicity to cells of the test systems used.
Full-text available
The objective of this study was to evaluate the antiproliferative, cytotoxic and genotoxic potential of salty liquid synthetic flavorings of Butter, Cheddar Cheese and Onion. The antiproliferative potential (2.9-1500 µg/mL) was assessed by MTT assay after 72h using the human tumor lines SF-295 (glioblastoma), OVCAR-8 (ovarian), HCT-116 (colon) and HL-60 (promyelocytic leukemia) and primary cultures of murine Sarcoma 180 (S180) and peripheral blood mononuclear cells (PBMC). Allium cepa bulbs were exposed to growing respective doses (1 mL and 2 mL). Only Butter and Cheddar flavorings revealed cytotoxic activity on cancer cells, with IC50 values ranging from 125.4 µg/mL (Cheddar - HCT-116) to 402.6 µg/mL (Butter - OVCAR-8). Butter flavoring was the most cytotoxic on PBMC (136.3 µg/mL) and increased cell division rate in relation to the mitotic index but did not cause cellular aberrations. Onion and Cheddar flavorings reduced the mitotic index after 24h and 48h exposure, but only Onion flavoring resulted in cellular aberrations and mitotic spindle abnormalities, such as anaphase and telophase bridges, micronucleated cells, conchicine-metaphases and amplifications. So, Butter, Onion and/or Cheddar flavorings caused significant changes in the division of meristematic cells of A. cepa and presented cytotoxic action even on decontrolled proliferating human tumor cells.
Full-text available
The objective of this study was to evaluate the action of Hymenaea stigonocarpa bark hydroalcoholic extract against a mutagenic compound using A. cepa meristematic root cells as a test system. The treatment groups were: Negative Control (NC) - distilled water; Positive Control (PC) - paracetamol at a concentration of 0.008 mg/mL, Jatoba Control (JC) - aqueous fraction jatobá-do-cerrado at 0.5 or 1.0 or 1.5 mg/mL, and Simultaneous Treatment (ST) -jatobá-do-cerrado aqueous fraction at a concentration of 0.5 or 1.0 or 1.5 mg/mL associated with paracetamol solution at a concentration of 0.008 mg/mL. All groups were analyzed at 24 and 48 h. Five onion bulbs (five replications) were used for each treatment group. The root tips were fixed in Carnoy and slides prepared by the crush technique. Cells were analyzed throughout the cell cycle, totaling 5,000 for each treatment group at each exposure time. Mitotic indices were subjected to statistical analysis using the chi-square test (p<0.05). From the results it was found that the ST group, at the three concentrations, significantly potentiated the antiproliferative effect of the test system cells when compared to PC, NC and TJ at the three concentrations. Furthermore, the three ST concentrations significantly reduced the number of cell aberrations when compared to the number of aberrant cells obtained for the PC, demonstrating antimutagenic action on the A. cepa test system cells.
Full-text available
Abstract Powdered juices are widely consumed by the population especially because of their convenient preparation, availability in various fruit flavors and low cost when compared to other industrialized beverages. They have complex formulation, consisting of several classes of food additives. However, there are no scientific studies on the toxicity of these foods. Thus, this study evaluated the toxicity at the cellular level of industrialized powdered juices of orange and guava flavors of three different food companies. This analysis was made using root meristem cells of Allium cepa L., at the exposure times of 24 and 48 hours, and two concentrations, 30 g/1000 mL, considered ideal for consumption according to the label of the products, and 30 g/500 mL. Both flavors of juices, of the three companies, in both concentrations and the two exposure times promoted significant antiproliferative effect to root meristem cells and caused a statistically significant number of mitotic spindle changes and micronuclei in cells of the test system used. Therefore, under the studied conditions, all the samples of juice powder exhibited cytotoxic and genotoxic potential.
Full-text available
The objective of this work was to evaluate the cytotoxic effect of the food dyes erythrosine, brilliant blue and red 40 on the cell cycle of Allium cepa L. Each dye was evaluated at doses of 0.4 and 4.0 ml, at exposure times of 24 and 48 hours, in onion root tip cells. Cells and the presence of chromosomal aberrations were analyzed throughout the whole cell cycle, totaling 5,000 cells for each group of bulbs. The mitotic index was calculated and the statistical analysis was conducted through the Chi-square test (p < 0.05). From the obtained results, it was verified that the food additives erythrosine and brilliant blue were not cytotoxic to the cells of the test system. However, the red 40 dye, at the two evaluated doses and the two exposure times used in this bioassay have promoted a significant reduction in cell division and induced the emergence of anaphasic and telophasic bridge aberrations and micronucleated cells. Additional cytotoxicity studies should be conducted to add information to these and other previously obtained results in order to evaluate, with property, the action of these three dyes on a cellular level.
Full-text available
Based on the concentration of Malathion used in the field, we evaluated the genotoxic potential of low concentrations of this insecticide on meristematic and F 1 cells of Allium cepa and on rat hepatoma tissue culture (HTC cells). In the A. cepa, chromosomal aberrations (CAs), micronuclei (MN), and mitotic index (MI) were evaluated by exposing the cells at 1.5, 0.75, 0.37, and 0.18 mg/mL of Malathion for 24 and 48 hr of exposure and 48 hr of recovery time. The results showed that all concentrations were genotoxic to A. cepa cells. However, the analysis of the MI has showed non-relevant effects. Chromosomal bridges were the CA more frequently induced, indicating the clastogenic action of Malathion. After the recovery period, the higher concentrations continued to induce genotoxic effects, unlike the observed for the lowest concentrations tested. In HTC cells, the genotoxicity of Malathion was evaluated by the MN test and the comet assay by exposing the cells at 0.09, 0.009, and 0.0009 mg/5 mL culture medium, for 24 hr of exposure. In the comet assay, all the concentrations induced genotoxicity in the HTC cells. In the MN test, no significant induction of MN was observed. The genotoxicity induced by the low concentrations of Malathion presented in this work highlights the importance of studying the effects of low concentrations of this pesticide and demonstrates the efficiency of these two test systems for the detection of genetic damage promoted by Malathion.
Full-text available
Este estudo teve por objetivo avaliar o efeito citotóxico dos corantes alimentares amarelo crepúsculo, vermelho bordeaux e amarelo tartrazina sobre o ciclo celular de Allium cepa L. Cada corante foi avaliado nas doses de 0,4 e 4,0 mL, nos tempos de exposição de 24 e 48 horas, em células de pontas de raízes de Allium cepa L. Prepararam-se lâminas e analisaram-se células, em todo o ciclo celular, e a presença de aberrações celulares, totalizando 5.000 células para cada dose avaliada. Foi calculado o índice mitótico e a análise estatística foi feita por meio do teste quiquadrado (p < 0,05). Os resultados mostraram que os três corantes, nas doses e tempos de exposição avaliados, foram citotóxicos às células do sistema-teste utilizado. Estudos adicionais de citotoxicidade devem ser conduzidos para se somar a estes resultados e, assim, avaliar, com propriedade, a ação destes três corantes em nível celular.