Natural Product Research

Formerly Natural Product Letters

ISSN: 1478-6419 (Print) 1478-6427 (Online) Journal homepage: https://www.tandfonline.com/loi/gnpl20

Genotoxic evaluation, secondary metabolites and

antioxidant capacity of leaves and roots of Urera
baccifera Gaudich (Urticaceae)

A.L. Gindri, L.B. de Souza, R.C. Cruz, A.A. Boligon, M.M. Machado & M.L.
Athayde

To cite this article: A.L. Gindri, L.B. de Souza, R.C. Cruz, A.A. Boligon, M.M. Machado & M.L.
Athayde (2014) Genotoxic evaluation, secondary metabolites and antioxidant capacity of leaves
and roots of Urera�baccifera Gaudich (Urticaceae), Natural Product Research, 28:23, 2214-2216,
DOI: 10.1080/14786419.2014.920333

To link to this article: https://doi.org/10.1080/14786419.2014.920333

View supplementary material Published online: 11 Jun 2014.

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Natural Product Research, 2014
Vol. 28, No. 23, 2214–2216, http://dx.doi.org/10.1080/14786419.2014.920333

SHORT COMMUNICATION
Genotoxic evaluation, secondary metabolites and antioxidant capacity of
leaves and roots of Urera baccifera Gaudich (Urticaceae)
A.L. Gindria, L.B. de Souzab, R.C. Cruza, A.A. Boligona, M.M. Machadoc and M.L. Athaydea*
a
Post Graduate Program in Pharmaceutical Sciences, Phytochemical Research Laboratory, Department of
Industrial Pharmacy, Federal University of Santa Maria, Build 26, room 1115, Santa Maria, RS 91105-
900, Brazil; bPost Graduate Program in Agrobiology, Biology Department, Federal University of Santa
Maria, Santa Maria, Brazil; cPost Graduate Program in Pharmaceutical Sciences, Federal University of
Pampa, Uruguaiana, Brazil
(Received 2 November 2013; final version received 29 April 2014)

In addition to phenolics, flavonoids, flavonols, alkaloids and condensed tannins, our

tests identified the antioxidant and genotoxic properties in the crude extract (CE) and
fractions of Urera baccifera (Urticaceae) roots and leaves. Oxalic acid (OA) content
was determined by HPLC-DAD, which presented high values in the roots (1.82 ^ 0.21,
1.79 ^ 0.22 and 1.38 ^ 0.15 mg/g in butanolic, CE and ethyl acetate fraction,
respectively). OA caused a 30.7% reduction in the leucocyte proliferation, followed by
butanolic fractions of roots (24.15%) and leaves (23.28%). The mitotic index was lower
in butanolic fractions of leaves (8.7%) and roots (8.3%), similar to the OA index, which
was 6.0%. The DNA damage index in cultured leukocytes was observed for OA (19.33)
and butanol fraction treatments (22.67 and 16, respectively, for leaves and roots).
Antioxidant capacity (DPPH and TBARS) was moderated, which was confirmed by the
low phenolic, flavonol and flavonoid contents in both parts of the plant.
Keywords: HPLC-DAD; DPPH; oxalic acid; phenolic compounds; TBARS;
genotoxicity

1. Introduction
Urera baccifera (L.) Gaudich. ex Wedd., from Urticaceae, is popularly known as ‘Urtigão’ in
Brazil and infusions of its leaves and roots are used to treat skin and urinary infections. In Costa
Rica, the leaves are used against inflammatory conditions, and it exhibits good results in anti-
inflammatory tests when injected intraperitoneally (Badilla et al. 1999).
Due to the very few studies on the chemical constitution and antioxidant capacity of
U. baccifera, this study aimed to quantify the polyphenols, flavonoids, flavonols, condensed
tannins and alkaloids by using spectrophotometric methods. In addition, the lipid peroxidation
inhibition in blood samples and the ability to scavenge the DPPH radical were used to evaluate
the antioxidant capacity of the crude extract (CE) and fractions obtained from the plant’s leaves
and roots. Genotoxic activity was evaluated using oxalic acid (OA) as reference, which was also
quantified by using HPLC.

2. Results and discussion

The contents of polyphenols, flavonoids, flavonols, condensed tannins and alkaloids (Table S1)
presented significant differences between the extracts and fractions of leaves and roots

*Corresponding author. Email: margareth@smail.ufsm.br

q 2014 Taylor & Francis

 Natural Product Research 2215

( p , 0.05). Previous studies on the root’s CE by Gindri et al. (2010) found very similar results
for polyphenols (29.76 ^ 1.5 mg/g), flavonoids (16.42 ^ 0.1 mg/g) and alkaloids
(1.58 ^ 0.02 mg/g of plant extract). According to the Mannion and Menezes (2010) study,
the CE and fractions from leaves of Urera collected in São Paulo State (Brazil) presented results
varying from 0 to 77.75 mg of gallic acid/g dry material, with the EtOAc fraction displaying the
highest value. The same fraction showed higher contents of flavonoids, where the values ranged
from 0 to 27.14 mg/g of rutin equivalents. The differences between all these contents may arise
due to the different places of sample collection, soil and climate, among other factors. Alkaloids
and condensed tannins were quantified in the CE of Urtica urens displaying 0.58 ^ 0.03 and
2.92 ^ 0.38 mg/g, respectively (Jimoh et al. 2010). Previously, the alkaloid contents
(1.58 ^ 0.2 mg/g) were determined in U. baccifera by Gindri et al. (2010), and the results
were at least three times higher than those obtained by Jimoh using the same methodology.
Alkaloid contents (1.6 ^ 0.04 mg/g) found in this study were very similar to those determined in
2010, showing no statistical differences.
The antioxidant activity with the DPPH photometric assay (Table S2) shows the IC50
(inhibition concentration of 50% oxidant activity) in mg/mL and the percentage of inhibition of
DPPH scavenging at the highest concentration of the extracts used (250 mg/mL). In general, the
plant was not able to scavenge the DPPH radical. In addition, no positive relationship was found
between the polyphenol content and the scavenging activity when using the DPPH method. For
example, the EtOAc and BuOH fractions from the leaves, whose phenolic contents do not differ,
presented very different IC50 and percentage of DPPH scavenging at 250 mg/mL; in the same
way, CE and CHCl3 fraction from the roots, whose contents of polyphenolics were statistically
different, presented very similar values of IC50 and percentage of DPPH scavenging at 250 mg/
mL (Tables S1 and S2). Taking these data together, we suggest that the moderated antioxidant
effect of U. baccifera may be due to other secondary metabolites. In the DPPH test, the CE of the
roots of U. baccifera exhibited a similar IC50 value, 188.57 mg/g extract (Gindri et al. 2010),
supporting the results obtained here.
The results of lipid peroxidation are expressed in nmol of malondialdeyde/mL erythrocyte
(Table S2). In this test, only a slight variation between the results of leaves and roots was
observed. Leaf extracts presented a slightly lower value; however, there were no significant
differences ( p . 0.05) among the root samples and among the leaf samples, except when the
two parts of the plant were compared. In contrast, Özen et al. (2010), after testing the
hydroalcoholic extracts of roots, flowers, leaves and seeds of Urtica pilulifera in total
thiobarbituric acid-reactive substances assay and peroxidation of linoleic acid assay, concluded
that all extracts had high antioxidant activities.
The OA quantification displayed higher results from the roots when compared with the
leaves (Table S3). The OA and its soluble salts can be found in the vegetable tissues in
significant values, and can induce intoxication by oxalate in living beings (Noonan & Savage
1999). The CE and fractions of U. baccifera roots exhibited high values of OA, from four to six
times higher than in the leaves, and the BuOH fraction exhibited the maximum value. As
expected, CE and BuOH fractions furnished elevated levels, due to the polarity of these solvents.
There were significant differences between all the samples tested ( p , 0.05), except for the
BuOH fraction and CE of roots ( p ¼ 0.1972).
The results of the determination of the effect on the genetic material of leucocytes in culture
are shown in Figures S1– S3. Figure S1(A) shows the effects of U. baccifera in the proliferation
of leucocyte cultures. OA caused a 30.7% reduction in the leucocyte proliferation. The samples
showing similar effects were the BuOH fractions of the roots (24.15% of reduction) and leaves
(23.28% of reduction). OA caused 15% leucocyte deaths, as shown in Figure S1(B). The BuOH
fraction of the leaves and roots displayed the highest values in this assay (7.0% and 5.0%,
respectively), followed by the CE (2.33% in both).
2216 A.L. Gindri et al.

The mitotic index in the leucocyte culture was lower in the BuOH fraction of the leaves and
roots (8.7% and 8.3%, respectively), similar to the OA index, which was 6.0%, considering the
negative control (Figure S2(A)). However, there were no statistically significant differences
between these treatments. The production of micronuclei was higher in the OA treatment
(7 micronuclei/100 cells) and significantly different from all the treatments, as shown in Figure
S2(B). Once again, the BuOH fraction presented similar results, with a median of 3 micronuclei
in the roots and 2.7 in the leaves, although no statistical differences were observed between these
fractions and the other fractions of the plant or even the negative control. The DNA damage
index in cultured leucocytes (Figure S2(C)) was observed in the OA treatment (19.33). The same
effect was observed in the BuOH fraction of the leaves (22.67) and roots (16.0), although no
significant differences were observed between the OA and U. baccifera fractions.
Taking into consideration the genotoxicity assays, the BuOH from roots and leaves
presented the highest quantities of OA and displayed results comparable with the standard,
indicating that this compound can be responsible for the toxic effects of the plant. Conversely,
the CE had similar quantity of this acid and the damage in leucocytes was not significantly
elevated, which indicates that more metabolites could be involved in the genotoxic effects of the
BuOH fractions or there are some compounds in the CE which could inhibit or prevent this
genotoxicity.

3. Conclusion
The leaves and roots of U. baccifera present a moderate antioxidant capacity that was confirmed
by the low polyphenol, flavonoid and flavonol contents. However, the plant presented a
significant quantity of OA, which could explain its stinging effect. Similarly, the genotoxic
effects of the CEs and fractions in the leucocyte culture may be related to the presence of OA in
the plant.

Supplementary material
Supplementary material relating to this article is available online, alongside Tables S1 –S3 and
Figures S1 and S2.

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