204
ISSN: 2763-5724 / Vol. 04 - n 04 - ano 2024
ACUTE TOXICITY OF GENERIC AND IN VITRO REFERENCE
TOXICAMOL USING THE ARTEMIA SALINE TOXICITY
BIOINDICATOR LEACH
Eduardo Lopes Carreiro de Alencar1
Ana Paula Guimarães Sá2
Renato Rego da Silva3
Jurandy do Nascimento Silva4
Abstract: This study aims to analyze, through an experimental study with a quantitative and
descriptive approach, the toxicity and identication of the 50% Lethal Concentration (LC50) of
the reference and generic paracetamol drops, both commercially available and produced in Brazil,
comparing them with each other. The lethality test was performed on Artemia salina according to
the methodology described by Meyer (1982), with some modications. When performing the toxicity
test on Artemia salina, for generic paracetamol, the LC50 obtained after 24 hours was 998.3 ± 0.2μg/
mL. The reference paracetamol presented LC50 of 612.2 ± 1.3μg/mL. Based on the analysis of this
research, it was found that the reference paracetamol presented signs of toxicity in nauplii of the
microcrustacean Artemia salina, presenting a LC50 ≤1000μg/mL. Generic paracetamol showed a
LC50 ≤1000μg/mL with a range above 1000 ug/mL indicating non-toxicity for this sample.
Keywords: Toxicity. Paracetamol. Artemia salina
1 Doctor, Pediatry resident-federal university of Piauí-UFPI
2 Pediatry resident-federal university of Piauí-UFPI
3 Doctor, Pediatry resident-federal university of Piauí-UFPI
4 PhD in Biotechnology at the Federal University of Piauí -UFPI
205
ISSN: 2763-5724 / Vol. 04 - n 04 - ano 2024
INTRODUCTION
Paracetamol or acetaminophene is a drug considered safe and eective, with analgesic and
antipyretic properties, with poor anti-inammatory action, to relieve light to moderate pain, being
eective for all age groups (BRAYNER, SILVAEALMEIDA, 2018). It is a low -cost and low -cost
drug (Mezarobba and Bitencourt, 2018).
Because it is a medicine that does not require medical prescription there is a high index of
indiscriminate use. Despite being considered safe and eective in its therapeutic doses, but when
consumed at doses higher than the recommendation it can cause hepatotoxicity. (Ribeiro et al. 2023).
Paracetamol -induced poisoning happens more frequently in young people 15 to 24 years old,
individuals who make abuse of alcohol and individuals aged 40 years of 40 years occur at high risk,
the hepatic failure and may be fatal, and may be fatal. (SILVA JUNIOR et al; 2019).
There are several pharmaceutical presentations sold, among them: tablets, oral solution,
suppositories and injectables (RIBEIRO et al, 2023). Due to ease and broad use in Brazil, it is important
to evaluate the pharmacological patterns of acetomiphene. The variability of laboratories, forms of
presentation and classication of the drug (reference, generic, similar and manipulated), suggests a
diversity in the quality standard of this, and it is necessary to evaluate such criteria to avoid possible
health damage (Teixeira, 2016).
In Brazil, according to Law 9,787 of February 10, 1999, the drugs considered “reference”
are innovative products whose eectiveness, safety and quality were scientically proven with the
competent federal agency, at the time of registration. Upon protection period of the reference product
patent, generic and similar drugs can be produced that, similar to the reference, follow the principles
of bioequivalence and bioavailability to obtain the same therapeutic eect for later records and
commercial use (. (Brazil, 1999; .Lopes; Costa, 2016, Garcez et al., 2019).
Soline artemia is a species of microcrustacean of the order years, found in Salgadas waters
(Carvalho et al., 2008). It is used as a living food for sh, and its eggs are found in aquarizers stores and
206
ISSN: 2763-5724 / Vol. 04 - n 04 - ano 2024
is widely used in toxicological studies due to low cost and easy cultivation. Several works have been
trying to correlate toxicity over A. saline with antifungal, viruscid, antimicrobial, tripanosomicide
and parasiticide activities. Lethality tests are done in toxicological tests and the median lethal
concentration (Cl50) indicates death in half a sample (BEDNARCZUK, 2010).
Thus, this work aimed to evaluate the toxicity of the acetaminopus drops: generic and
reference, in A. saline model.
METHODOLOGY
This work resulted from an experimental study, with quantitative and descriptive approach,
which evaluated toxicity and identied the lethal concentration 50% (CL50) of acetaminophen (oral
solution) reference and generic, both commercially available and produced in Brazil, comparing them
each other.
A. saline eggs were placed to hatch in an articial marine water solution (NaCl 77.23%,
MGSO4 9.62%, MGCl 7.13%, CACL23.32%, KCl 2.1%and NAHCO3 0, 59%) and after 48 hours, ten
nauplies of saline artemia were transferred with a micropipette to testing tubes containing articial
marine water and reference and generic paracetamol samples that were diluted in serial concentrations
(125µg/mL, 250 µg/ml, 500 µg/ml, 750 µg/ml and 1250 µg/ml).
The lethality test was held in Artemia Salina, cultivated at the Food Analysis/Sanitation
Analysis Laboratory of the Federal Institute of Piauí-IFPI/CTZS, according to the methodology
described by Meyer (1982), with some modications.
The specimens were distributed in three specimens for each concentration of brand and
generic acetaminophen, as well as negative control with articial marine water and positive control
with 0.0006 mg/ml of copper sulfate (COSO4).
The tubes were left at room temperature for 24 hours. After this period, the number of
surviving cassaways was held. They were observed near a light source and considered alive all those
207
ISSN: 2763-5724 / Vol. 04 - n 04 - ano 2024
who had any kind of movement when subjected to a slight stirring.
The number of living naupli in relation to the increase in the concentration of extracts
and fractions was used to calculate the values of the CL50. Thus, given the values of the CL50
obtained through this method, substances tested in dierent concentrations are classied as toxic
when the Cl50 is less than 1000 µg/ml and non -toxic when the Cl50 is greater than 1000 µg/ml.
SUBMITTED AND DISCUSSION
The results were expressed CL50 obtained by the statistical method of nonlinear regression
followed by the 95% condence interval (GraphPadPrism 6.0®).
Table 1 - Lethal Concentration 50% (Cl50) of the generic acetaminophen and the reference paracetamol
after 24 hours of exposure by the toxicity test with A. saline
A comparison between generic and reference paracetamol performed by Lima (2019) against
A. Salina found a CL50 for the generic acetaminopus of 265.377 μg/ml with condence interval
within 258,223 to 272,060 μg/mL and for the reference paracetamol of 252.578 μg/mL with a break
within 249,600 to 255.619 μg/mL that were distant from the CL50 found for the generic paracetamol
and reference in the present study that were 998.3 ± 0.2 μg/ml and 612.2 ± 1 , 3 μg/ml, respectively.
Garcez et al. (2018) performed a toxicity bioassay with A. saline using the same method
with generic and reference dipyrone and obtained a CL50 for generic dipyrone of 986.9 μg/
mL and 654.1 μg/mL for reference dipyrone . Results close to those obtained in this study.
208
ISSN: 2763-5724 / Vol. 04 - n 04 - ano 2024
Conclusions
The reference paracetamol showed indications of toxicity in microcrustacean naúplies
artemia saline with a cl50 ≤1000µg/ml. The generic acetaminophen showed a cl50 ≤1000µg/ml with
an interval above 1000 Ug/ml indicating atxicity for this sample.
Thus, it is possible to carry out other deeper studies, comparing the generic acetaminophen
with the reference.
Bibliographic references
BEDNARCZUK, V.O. et al. Testes in vitro e in vivo utilizados na triagem toxicológica de produtos
naturais. Visão Acadêmica. Curitiba, v. 11, n. 2, p. 44, jul/dez 2010.
BRAYNER, N. F.; SILVA, A. A. da; ALMEIDA, F. R. de. O risco do uso irracional do paracetamol
na população brasileira e seus efeitos na hemostasia. Revista Cientíca da FASETE, v. 2018.1, p.138-
153, 2018.
BRASIL. Lei n.º 9.787, de 10 de fevereiro de 1999. Altera a Lei n.º 6.360, de 23 de setembro de 1976,
que dispõe sobre a vigilância sanitária, estabelece o medicamento genérico, dispõe sobre a utilização
de nomes genéricos em produtos farmacêuticos e dá outras providências. Diário Ocial da União,
Brasília, DF, 11 fev. 1999. Seção 1, p. 4.
CARVALHO, C. A. de. et al. Cipó-cravo (Tynnanthus fasciculatus miers- Bignoniaceae): Estudo -
toquímico e toxicológico envolvendo Artemis salina. Revista Eletrônica de Farmácia Vol 6(1), 51-58,
2009. GARCEZ, B. B. D.. et al. Toxicidade aguda da dipirona sódica in vitro utilizando o bioindica-
dor de toxicidade Artemia salina Leach. Revinter, v. 11, n. 2, p. 114-119, jun. 2018.
LIMA, Leticia Mirelle Vieira. Avaliação toxicológica das apresentações de ácido acetilsalicílico e pa-
racetamol frente à Artemia salina Leach. 2019. 42 f. (Trabalho de Conclusão de Curso – Monograa)
– Curso de Farmácia, Universidade Federal de Campina Grande, Cuité – PB, 2019.
209
ISSN: 2763-5724 / Vol. 04 - n 04 - ano 2024
LOPES, J. C.; COSTA, T. M. F. Controle de qualidade de soluções orais de dipirona sódica 500 mg/
mL referência, genérico e similar comercializadas em Gurupi-TO. AMAZÔNIA: SCIENCE & HE-
ALTH, v. 4, n. 4, p. 03-11, 2016.
MEZAROBBA, G.; BITENCOURT, R. M. de. Toxicidade do paracetamol: o álcool como um fator
de risco. Unoesc & Ciência - ACBS, [S. l.], v. 9, n. 1, p. 105–112, 2018. Disponível em: https://perio-
dicos.unoesc.edu.br/acbs/article/view/12352. Acesso em: 12 ago. 2024.
RIBEIRO, B. V. et al. . Os riscos do uso indiscriminado e irracional do paracetamol. Brazi-
lian Journal of Implantology and Health Sciences , [S. l.], v. 5, n. 3, p. 1016–1035, 2023. DOI:
10.36557/2674-8169.2023v5n3p1016-1035. Disponível em: https://bjihs.emnuvens.com.br/bjihs/ar-
ticle/view/349. Acesso em: 12 ago. 2024.
SILVA JÚNIOR, J. G . et al. Hepatotoxicidade induzida pelo paracetamol e a utilização do nomogra-
ma de rumack-matthew para avaliar a terapêutica com n-acetilcisteína. Revista Uningá, [S. l.], v. 56,
n. 4, p. 65–84, 2019. DOI: 10.46311/2318-0579.56.eUJ2087. Disponível em: https://revista.uninga.
br/uninga/article/view/2087. Acesso em: 12 aug. 2024.
TEIXEIRA, D. A., et al. Avaliação do controle de qualidade da dipirona sódica: de referência, similar,
genérica e manipulada, comercializadas no municipio de Teólo-Otoni, mg. Revista Multidisciplinar
do Nordeste Mineiro –Nov. 2016. Disponível em: < http://www.unipacto.com.br/revista2/arquivos_
pdf_revista/revista2016_2/9.pdf>. Acesso em 10/08/2024.
