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Sistema de Información Científica
Red de Revistas Científicas de América Latina y el Caribe, España y Portugal
Rev. Cubana Quím.
Vol. 29, no.3, sept-dic, 2017, págs. 330-340, e-ISSN: 2224-5421
Composición química y actividad larvicida de un extracto de
marañón frente a larvas de mosquitos
Chemical composition and larvicidal activity of cashew nutshell ethanolic
extract against mosquito larvae
Dr. C. Jesús Rafael Rodriguez Amado
I
; Dr. C. Raimundo Nonato Picanço Souto
II
;
MSc. Miriam Santos Magalhães
II
; Dr. C. Julio César Escalona Arranz
III
;
Dr. C. Jose Carlos Tavares Carvalho
I
jiribilla2009@gmail.com
I
Laboratório de Pesquisa em Fármacos. Faculdade de Ciências da Saúde. Universidade
Federal do Amapá, Macapá, AP. Brasil;
II
Laboratório de Arthropoda. Faculdade de
Biologia. Universidade Federal do Amapá, Macapá, AP. Brazil;
III
Departament of Pharmacy,
University of Oriente, Santiago de Cuba, Cuba.
Recibido: 13 de febrero de 2017
Aprobado: 24 de abril de 2017
Resumen
El objetivo de este trabajo es evaluar la composición química, así como la actividad larvicida
contra mosquitos
Aedes Aegypti
y
Culex quinquefasciatus
de un extracto de la cáscara de nuez
de
Anacardium Occidentale L.
o marañón. Con esta intención, un extracto hidro-etanólico es
preparado por maceración y en proporción de 1 g de corteza seca en 3 ml de etanol al 70 %. Al
extracto
se
le
determinó
la
presencia de compuestos
fenólicos,
cumarinas,
quinonas,
antocianidinas, triterpenos, esteroides, flavonoides y saponinas. La actividad larvicida mostró
una concentración letal media (LC50) de 35,81 ppm y 40,21 ppm para larvas de
A. aegypti
(cepas Macapá y Rockefeller respectivamente) y 21,47 ppm para
C. quinquefasciatus.
La LC50
para la actividad anticolinesterásica fue de 43,27±2,22 ppm, pudiendo asociarse la actividad
larvicida del extracto a su capacidad inhibitoria sobre esta enzima.
Palabras clave:
actividad larvicida, extracto etanólico de marañón, mosquitos.
Abstract
The goal of this work is to evaluate the chemical composition and the larvicidal activity
of an extract from the nutshell (
Anacardium occidentale L
.) against
Aedes aegypti
and
Culex quinquefasciatus
larvae. A Hydro-ethanolic extract was prepared by maceration using 1 g
of cashew nutshell in 3 mL of ethanol 70 %. The larvicidal effect and the acetylcholinesterase
inhibitory
activity
of
the
extract
were
evaluated.
The
extract
content
includes
phenolic compounds, coumarins, quinones, anthocyanidins, triterpenes, steroids, flavonoids,
saponins, lipid compounds, aliphatic acids, and terpenoids. The larvicidal activity of the extract
(LC50) for
A.aegypti
was 35,81 ppm and 40,21 ppm for
A. aegypti
(Macapá and Rockefeller
strains), and 21,47 ppm for
C.quinquefasciatus
. The IC50 for the AChE inhibitory test was
43,27±2,22 ppm. The evaluated extract presents a good AChE inhibitory activity and a potent
larvicidal activity. The larvicidal effect could be associated with the AChE inhibitory activity of
the extract.
Keywords
: larvicidal activity, nutshell ethanolic extract, mosquitoes.
Composición química y actividad larvicida de un extracto de marañón frente
a larvas de mosquitos
Rev. Cubana Quím., vol. 29, no. 3 septiembre-diciembre, 2017. e-ISSN 2224-5421
331
Introducción
The synthetic chemical insecticides are the primary option to combat adult mosquitoes
and immature forms of
Aedes aegypti, Anopheles sp. and Culex quinquefasciatus
. These
mosquitoes are vectors of human diseases such as Dengue, Chikungunya, and Zika
viruses
(
A.aegypti
),
malaria
(Anopheles),
filariasis
and
viral
encephalitis
(
C. quinquefasciatus)
[1]. Brazil and Cuba are among the countries where dengue is
endemic. The morbidity of dengue in Brazil grows up 48 percent until May 2016, while
Zika virus is now a world pandemic. Brazil has the highest number of cases of Zika
virus around the world, followed by Colombia and Venezuela [2].
It
was
reported
the
appearance
of
resistance
of
mosquitoes
to
some
chemical
insecticides
like
DDT
(an
organochloride)
[3],
Malathion,
Diazinon
and
others
organophosphates, especially in some populations of Brazil, Puerto Rico, and others
Caribbean countries [4]. Additionally, a decrease in mosquitoes' susceptibility to the
activity of the pyrethroids has been observed [5]. That is why it is urgent to find new
alternatives for mosquito control, different from the classical chemical insecticides.
In this sense, natural products can be an eco-friendly source of compounds with
bioactivity against mosquitoes and others vectors. Many secondary metabolites have
been tested as alternatives to vector control, demonstrating their larvicide activity in
different ways [6]. In this context, several studies to discover vegetal substances for this
approach have been performed [7].
Literature refers that some species of the Anacardiaceae family prove a larvicide
activity. This family contains about 600 species distributed through 73 genera, including
the genus Anacardium, which has eight species [8]. A. occidentale, popularly known as
cashew (Cajú, in Brazil and Marañón in Cuba), is distributed around the world in
tropical zones. The bark and leaves contain anti-inflammatory and astringent agents,
which are used to prepare a decoction considered effective in the treatment of diarrhea,
diabetes, eczema. It is also used as mouthwash [9].
The cashew nutshell oil, known as CNSL (English acronym for cashew nutshell liquid)
is obtained by heating the cashew nutshell to 70 °C. Anacardic acids (four isomers) are
phenolic compounds biosynthesized from fatty acids and represent about 70 % of the
CNSL present in the shell of the cashew nutshell [10]. These compounds have a potent
larvicidal activity, but no mechanism has been described yet [11]. This extraction
process can generate some new metabolites or “artefacts” product of the heat applied.
Jesús Rafael Rodriguez Amado; Raimundo Nonato Picanço Souto; Miriam Santos Magalhães;
Julio César Escalona Arranz; Jose Carlos Tavares Carvalho
332
Rev. Cubana Quím., vol. 29, no. 3 septiembre-diciembre, 2017. e-ISSN 2224-5421
On the other hand, maceration with ethanol is a classical method that allows the
extraction of the secondary metabolites with high affectivity and low cost. At the best of
our knowledge, there are not studies of the ethanolic extract of the cashew nutshell
of
Anacardium occidentale
L within this purpose. That is why the aim of this work was
to determine the chemical composition of a hydro-ethanolic extract of cashew nutshell
of
Anacardium occidentale
L (
Anacardiaceae)
and evaluate it in their larvicidal activity
against
Aedesa egypti
(two strains) and
Culex quinquefasciatus
larvae, as well as the
AChE inhibitory effect.
Materials and methods
Plant collection:
The fruit of
A. occidentale
was collected in the morning time at the
municipality of Macapá, Amapá, Brazil (030 380N / 498 820W) in October 2015.
Dr. Patrick Canterbury makes the plant identification and a voucher specimen with the
register number 018781 was deposited at the Amapá Herbarium (HAMAB), Macapá,
Amapá, Brazil.
Preparation of the cashew nutshell extract:
The cashew nutshell was separated from
the fruit and cut, longitudinally, with steel knife in order to remove the core.
The cashew nutshell was washed with distilled water for mechanical separation of
impurities. One gram of cashew nutshell was extracted by Maceration (72 h) in an
amber vessel using a proportion of three milliliters of ethanol 70 % for each gram of
drug. Every day, the extract was manually stirred. After 72 h the extract was filtered and
stored at room temperature (25 ºC).
Physical properties of the extract:
The pH of the 70 % hydro-ethanolic extract from
cashew nutshell was measured on a pH meter mPA210 (MS TECNOPON, Brazil).
The
equipment
was
calibrated
by
using
pH
7,0
(±0,02)
buffer
solutions
and
pH 4,0 (± 0,02) (Alphatec, Brazil). Measurements were performed in triplicate and
results were expressed as the mean and standard deviation.
Relative density was determined at 25 °C by using a pycnometer method [12]. A set
of 3 pycnometers (50 mL) and an analytical balance with accuracy to 0,000 1 g
(Acculab, Sartorius, Germany) were used. The analysis was performed in triplicate and
results were expressed as the mean and standard deviation.
The Total Soluble Substances contained in the cashew nutshell hydro-ethanolic extract
were determined by gravimetric method [12] with some modifications. In a porcelain
crucible, 5 mL of sample was placed and the mass was determined (CME) on an
Composición química y actividad larvicida de un extracto de marañón frente
a larvas de mosquitos
Rev. Cubana Quím., vol. 29, no. 3 septiembre-diciembre, 2017. e-ISSN 2224-5421
333
analytical balance (0,000 1g, Acculab, Sartorius, Germany). Afterwards, the crucible
with the sample was placed in a water bath at 100 °C until almost the liquid present was
removed. Then, the crucible was placed in an oven for one hour at 105 ºC. After this
period, the crucible with the residue was placed in a desiccator at room temperature, to
be weighed subsequently. The crucible was placed back in the oven following the same
procedure and after one hour is weighed again. The process continues until two
consecutive weight of the crucible with the residue (CMF) not differ more than 20 mg.
The assay was performed in triplicate. The Total Soluble Substances (TS) expressed in
percent was calculated using the equation 1:
% TS = [(CME- CMF)/CME]x100
(1)
Phytochemical screening:
Phytochemical screening was performed to evaluate the
presence of secondary metabolites by using the Chabra et al. methodology [13]. It was
tested for the presence of phenols, lipids substances, quinones, alkaloids, flavonoids,
triterpenes, steroids, saponins, and anthocyanidins.
Total phenols:
Total phenols content was obtained by Folin-Ciocalteu method at
760 nm [14]. Calibration curve was constructed using pyrogallic acid (Sigma-Aldrich,
USA) as external standard at 0,025, 0,050, 0,075, 0,100 and 0,125 mg/50 mL.
A spectrophotometer QUIMIS, AA6300 (Shimadzu, Japan) was used. The expression of
the calibration line was by the equation 2. Determinations were made in triplicate.
A = -0,007 8 + 1,824*C
(2)
r = 0,991 3 and r
2
= 0,982 7
where
A: absorbance
C: concentration expressed in mg/50 mL)
Evaluation of larvicidal activity:
Fourth-stage larvae of
A. aegypti
(Macapá and
Rockefeller strains) and
Culex quinquefasciatus
were used, which came from the
Arthropod
Laboratory
(ARTHROLAB),
Federal
University
of
Amapá.
The
experimental protocol was performed according to the World Health Organization
(WHO) [15]. One mL of the hydro-ethanolic extract to be tested was diluted in 99 mL
of distilled water to form a 1 % (as total solids) homogeneous solution.
Six experimental groups (for each species) using one of them as control and the other five
with a concentration of 10, 25, 50, 75, 100 ppm were performed. A total of 25 larvae per
Jesús Rafael Rodriguez Amado; Raimundo Nonato Picanço Souto; Miriam Santos Magalhães;
Julio César Escalona Arranz; Jose Carlos Tavares Carvalho
334
Rev. Cubana Quím., vol. 29, no. 3 septiembre-diciembre, 2017. e-ISSN 2224-5421
group were placed in the beaker. For the control group, in every case, 99 ml of distilled
water plus 1 mL of the solvent in which were prepared the extract (ethanol 70 %) was used.
All experiments were performed in triplicate and mortality was recorded after exposure
periods of 24 h.
Acetylcholinesterase (AChE) inhibitory activity:
The acetylcholinesterase inhibition
assay was developed as follow. An aliquot of the extract was dissolved in a base-tris
(0,05 M) buffer, following the Ellman methodology as described by Niño
et al.
[16].
Five levels of concentration (250 ppm, 125 ppm, 75 ppm, 50 ppm and 25 ppm) were
tested. Thus, 200 μL of acetylthiocholine iodide (15 mM, Sigma, USA), 1 000 μL
of Ellman's Reagent, 5,5'-Dithio-bis-(2-nitrobenzoic acid) (3 mM, Sigma, USA), and
200 μL of the extract solution were placed in the test tube. The reaction mix was
incubated for 25 min at 30 ºC. Then, the absorbance was measured (Shimadzu, Japan) at
412 nm. After that, 200 μL of AChE (0,3 U/mL) solution were added to start the
reaction and then the absorbance was read. A mixture of all components without the
extract was used as a control. The percentage of AChE inhibitory activity (% I
AChE
) was
calculated by the equation (3):
% I
AChE
= [(As - Ae) / As]x100
(3)
where
As
is the absorbance of the reactants without AChE enzyme
Ae
is the experimental absorbance after the reaction with AChE
All experiments were performed in triplicate. The IC
50
was determined by Probit
analysis using the StatGraphics Centurion.
Data analysis:
The software StatGraphics Centurion (StatEase Co. MA, USA) was
used for data analysis. Dosage-response (mortality) lines were estimated by using Probit
analysis. For all the estimations, 95 % confidence intervals (CI) were used.
Discussion of results
Preliminary characterization of the 70 % hydro-ethanolic extract
The
extract
showed
a
characteristic
odor,
with a
translucent
appearance
and
homogeneous dark brown color. The presence of particles in suspensions was not
observed. The physicochemical characterization of the extract is presented in table 1.
The phytochemical screening detects the presence of phenolic compounds, coumarins,
Composición química y actividad larvicida de un extracto de marañón frente
a larvas de mosquitos
Rev. Cubana Quím., vol. 29, no. 3 septiembre-diciembre, 2017. e-ISSN 2224-5421
335
quinones,
anthocyanidins,
triterpenes,
steroids,
flavonoids,
saponins
and
lipid
substances. The total phenol content was calculated in 5,52 μg/mL.
TABLE 1. PHYSICOCHEMICAL CHARACTERISTICS OF THE
HYDRO-ETHANOLIC EXTRACT (70 %) FROM
CASHEW OF (
A. occidentale
) NUTSHELL
Property
Unity
Mean
±
SD
Density (20 °C)
g/mL
0,910 ± 0,001
pH (1/ 15 mL of distilled water)
-
5,08 ± 0,03
Total polyphenols
%
5,52 ± 0,001
Total Soluble Substances
%
10,49 ± 0,17
Evaluation of larvicidal activity
The result of the larvicidal activity of cashew nutshell extract is presented in
table 2. The values of LC
50
and LC
90
with their limits at 95 % of confidence interval are
presented as well. The LC
50
and LC
90
for each species tested were statistically
significant (Data not shown). The R
2
value of the Probit analysis was greater than 85 %,
in all cases.
Jesús Rafael Rodriguez Amado; Raimundo Nonato Picanço Souto; Miriam Santos Magalhães;
Julio César Escalona Arranz; Jose Carlos Tavares Carvalho
336
Rev. Cubana Quím., vol. 29, no. 3 septiembre-diciembre, 2017. e-ISSN 2224-5421
TABLE 2. MORTALITY LEVELS INDUCED BY THE 70 % HYDRO-ETHANOLIC
EXTRACT FROM CASHEW (A
nacardium occidentale
L.) NUTSHELL ON LARVAE
OF
C. quinquefasciatus
,
A. aegypti
MACAPÁ, AND ROCKEFELLER STRAIN
Specie
Group
Concentration
Mean ± SD
Larvicidal activity
(95 % C.I., ppm)
Adjusted
R
2
(ppm)
(%)
LC50
LC90
A
aegyptiRockefeller
Strain
Control 0
0,00 ± 0,00
35,81
69,77
85,92
1
10,00
28,00 ± 5,00
(28,87-
43,03)
(60,82-
81,91)
2
25,00
46,67 ± 7,90
3
50,00
66,67 ± 2,90
4
75,00
93,33 ± 7,90
5
100,00
100,00 ± 0,00
A. aegyptiMacapá strain
Control 0
0,00 ± 0,00
40,21
76,25
87,50
1
10,00
22,67 ± 2,90
(34,04-
44,06)
(70,60-
82,91)
2
25,00
44,00 ± 5,00
3
50,00
62,67 ± 2,90
4
75,00
92,00 ± 0,00
5
100,00
100,00 ± 0,00
Culexquinquefasciatus
Control 0
0,00 ± 0,00
21,47
36,05
90,62
1
10,00
24,00 ± 5,00
(19,15-
23,97)
(32,79-
40,21)
2
25,00
40,00 ± 5,00
3
50,00
56,00 ± 5,00
4
75,00
91,00 ± 5,00
5
100,00
100,00 ± 5,00
AChE inhibition activity
It was noticed a concentration/activity relationship, making possible the calculation of
the IC
50.
The acetylcholinesterase inhibitory activity of the extract expressed as IC
50
was
estimated in 43,27 ± 2,22 ppm.
The extract presents a homogeneous and brilliant brown appearance. The pH value of
the extract (5,08 ± 0,03) can be attributed to the presence of secondary metabolites with
Composición química y actividad larvicida de un extracto de marañón frente
a larvas de mosquitos
Rev. Cubana Quím., vol. 29, no. 3 septiembre-diciembre, 2017. e-ISSN 2224-5421
337
acidic groups (phenolic compounds, anthocyanidins, and terpenoids) as well as free
fatty acid.
Total phenols content was estimated in 5,52 ± 0,01 μg/mL. This value is in concordance
with the reviewed literature when signed the phenolic compounds as the most abundant
metabolites in the Anacardium occidentale extracts [10]. The measured density for the
extract kept a close relation with the solvent ethanol 70 %. The total soluble substance
variable shows an acceptable level of the substances extracted.
Concerning the larvicidal activity of the extract in the three mosquitos larvae (adjusted
R
2
>85 %), it indicates that the observed mortality (Dependent variable) is plenty
explained by the variation of the extract concentration (Independent variable in the
Probit analysis).
The potential larvicidal application of natural products can be classified considering
mortality levels of larvae after 24-48 h of treatment with concentration of 250 ppm, as
follows: promising (mortality > 75 %), partially promising (50 % > mortality < 75 %),
weakly promising (25 > mortality < 50 %) and inactive (mortality < 25 %) [17].
In this context, the 70 % hydro-ethanolic extract of cashew nutshell can be declared as a
powerful larvicidal agent against the two mosquitoes’ larvae tested with IC
50
with
values five times lower that what is declared as promising larvicidal. It is noticeable that
a 100 % of mortality was achieved at the higher concentration tested in the three
different mosquito’s larvae. Therefore, this extract could have a great impact on the
vector of Dengue and Zika viruses in field experiments.
The inhibition of acetylcholinesterase is one of the mechanisms involved in the
insecticide and larvicidal activity of chemical and natural products [6]. A variety of
plants has been reported to show AChE inhibitory activity. Almost all the metabolites
detected in the 70 % hydro-ethanolic extract of cashew nutshell were reported for their
AChE inhibitory activity in previous reports [18]. The IC
50
estimated for this extract
was 43,27±2,22 ppm. This value can be considered good if compared to a variety of
natural compound reported by Suganthy
et al.
[18]. This fact suggests that the potent
larvicidal activity of this extract could be associated with their AChE inhibitory activity
as a possible action mechanism.
Jesús Rafael Rodriguez Amado; Raimundo Nonato Picanço Souto; Miriam Santos Magalhães;
Julio César Escalona Arranz; Jose Carlos Tavares Carvalho
338
Rev. Cubana Quím., vol. 29, no. 3 septiembre-diciembre, 2017. e-ISSN 2224-5421
Conclusions
The 70 % hydro-ethanolic extract of cashew nutshell shows a great variety of chemical
compound that can be responsible for the potent natural larvicidal activity demonstrated
against the three mosquito larvae tested. At the light of these results, this extract could be
used for larvae control of A. aegypti and C. quinquefasciatus in backyards and forest
zones. It is signed also that the larvicidal activity could be related to the strong AChE
inhibition. This work constitutes the first report in the chemical characterization
of a 70 % hydro-ethanolic extract of the Anacardiumoccidentale L. cashew nutshell and
their applicability as a potential larvicidal agent.
Acknowledgments
This work is supported by CNPq process No. 402332/2013-0. The authors specially
thank the support of the Universidade Federal do Amapá, Brazil, and CAPES.
References
1.
BARIK, K. T.; KAMARAJU, R.; GOWSWAMI, A. “Silica nanoparticle: a
potential new insecticide for mosquito vector control”.
Parasitology Research.
2012,
111
(3), 1075-1083. ISSN: 0932-0113
2.
WHO. Dengue: Data, maps and statistics of the PAHO/WHO. Number of
Reported Cases of Dengue and Severe Dengue (SD) in the Americas, by
Country: Deaths (SD/D) x100 CFR North America. Epidemiological Week
11.2016.
3.
DE ANDRADE, R. P. K.; RAILDA, A. R.; ALVES, A. M.; LIMA, A. C. C.;
MICHELLINE, S. E. “Insecticidal activity of the liquidof cashew nuts on
Aedes
aegypti
(Linnaeus,1762)
(Diptera:
Culicidae)
[In
Portuguese]”.
Revista
Brasileira de Biociências.
2013,
11
(4), 419-422. ISSN: 1980-4849
4.
KASAI, S.
et al.
“Mechanisms of Pyrethroid Resistance in the Dengue Mosquito
Vector,
Aedes aegypti
: Target Site Insensitivity, Penetration, and Metabolism”.
PLoS Neglected Tropical Diseases.
2014,
8
(6), e2948. ISSN: 1935-2735
5.
PEREIRA DA CUNHA, M.; LIMA, B. J. P.; BROGDON, V.; MOYA, E. G.;
VALLE, D. “Monitoring of resistance to the piretroids cypermethrin in Brazilian
Aedes aegypti
(Diptera: Culicidae) populations collected between 2001 and
Composición química y actividad larvicida de un extracto de marañón frente
a larvas de mosquitos
Rev. Cubana Quím., vol. 29, no. 3 septiembre-diciembre, 2017. e-ISSN 2224-5421
339
2003”.
Memórias do Instituto Oswaldo Cruz
.
2005,
100
(4), 441-444. ISSN:
0074-0276
6.
RATTAN, S. R. “Mechanism of action of insecticidal secondary metabolites of
plant origin”.
Crop Protection
.
2010,
29
(9), 913-920. ISSN: 0261-2194
7.
GUISSONI, P. A. C.; SILVA, G. I.; GERIS, R.; CUNHA, C. L.; SILVA, G. H.
H. “Larvicidal activity of
Anacardium occidentale
as an alternative to the
control
of
Aedes
aegypti
and
their
toxicity
in
Rattus
norvegicus
. [In
Portuguese]”.
Revista Brasileira de Plantas Medicinais
.
2013,
15
(3), 363-367.
ISSN: 1516-0572
8.
EIJNATTEN, M. C. L.
Anacardium occidentale
. In: Halvey, A.H. (Editor).
Handbook of flowering
. First edition. CRC Press, Boca Raton, Florida: USA;
2009. ISBN: 9780849339165
9.
NNAMANI,
V. C.;
OSAYI,
E. E.;
AAMA,
I. C.;
NWACHUKWU,
C.
“Larvicidal effects of leaf, bark and nutshell of
Anacardium occidentale
on the
larvae of
Anopheles gambiae
in Ebonyi State, Nigeria”.
Animal Research
International.
2011,
8
(1), 1353-1358. ISSN: 1597-3115
10.
HAMAD, B. F.; MUBOFU, B. E. “Potential Biological Applications of Bio-
Based
Anacardic
Acids
and
Their
Derivatives”.
International
Journal
of
Molecular Sciences
.
2011,
16
(4), 8569-8590. ISSN: 1422-0067
11.
FARNESI, C. L.
et
al. “
Physiological and Morphological Aspects of Aedes
aegypti
Developing
Larvae:
Effects
of
the
Chitin
Synthesis
Inhibitor
Novaluron”.
PLoS ONE.
2012,
7
(1), e30363. ISSN: 1932-6203
12.
BRAZILIAN PHARMACOPOEIA. Agencia de Vigilância Sanitária, ANVISA.
Brasil. 2010.
13.
CHABRA, S. C.; ULSO, F. C.; MSHIU, E. N. “Phytochemical screening of
Tanzanian medicinal plants (I)”.
Journal of Etnopharmacology.
1984,
11
(2),
157-179. ISSN: 0378-8741
14.
BRITISH PHARMACOPOEIA (BP). Her Majesty Stationary office. London,
UK. 2010.
15.
WHO. World Health Organization.
Guidelines for laboratory and field-testing of
mosquito larvicidal
. World Health Organization Communicable disease control,
Jesús Rafael Rodriguez Amado; Raimundo Nonato Picanço Souto; Miriam Santos Magalhães;
Julio César Escalona Arranz; Jose Carlos Tavares Carvalho
340
Rev. Cubana Quím., vol. 29, no. 3 septiembre-diciembre, 2017. e-ISSN 2224-5421
prevention and eradication. WHO pesticide evaluation scheme. 2005. Geneva,
Switzerland.
16.
NIÑO, J.; HERNÁNDEZ, J. A.; CORREA, Y. M.; MOSQUERA, O. M. “In
vitro inhibition of acetylcholinesterase by crude plant extracts from Colombian
flora”.
Memórias do Instituto Oswaldo Cruz
. 2006,
101
(7), 783-785. ISSN:
0074-0276
17.
MONTENEGRO, L. H. M.
et al.
“Terpenóides e avaliação do potencial
antimalárico, larvicida, anti-radicalar e anticolinesterásico de
Pouteria venosa
(Sapotaceae)”.
Revista Brasileira de Farmacognosia
. 2006,
16
, 611-617. ISSN:
1981-528X
18.
SUGANTHY, N.; KARUTHA, P. S.; PANDIMA, D. K. “Cholinesterase
Inhibitors from Plants: Possible Treatment Strategy for Neurological Disorders-
A Review”.
International Journal of Biomedicine and Pharmaceutical Sciences
.
2009,
3
, 87-103. ISSN: 1752-3788.
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