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REVISTA DE BIOLOGIA E CIÊNCIAS DA TERRA ISSN 1519-5228 
36 
Volume 14 - Número 2 - 2º Semestre 2014 
RESIDUES OF ENROFLOXACIN AND CIPROFLOXACIN IN POULTRY TISSUES 
Arnildo Korb1; Eleusis R. de Nazareno2; Paulo R. Dalsenter3; Ciro A. de O. Ribeiro4; Constança Pomba5 
ABSTRACT 
We investigated the existence of residues of the antimicrobials enrofloxacin and ciprofloxacin in 
broilers, which are exposed to the consumer in the metropolitan city Curitiba, Brazil. Antimicrobial 
residues in animal tissues, above the minimum allowed concentration, pose risks to human health. In 
10 samples analyzed by Liquid Chromatography (Mass Spectometry), at the Lanagro laboratory, from 
Ministry of Agriculture, Livestock and Supply of Brazil, considering a lower limit of 20 μg.kg-1 and 
maximum 100 μg.kg-1, no residues of ciprofloxacin and enrofloxacin were detected. These results 
showed that in the broiler samples of different brands, these antimicrobials were not detected. 
Keywords: Antimicrobial residues, poultry tissues, ciprofloxacin and enrofloxacin. 
RESÍDUOS DE ENROFLOXACINA E CIPROFLOXACINA EM TECIDOS DE FRANGOS 
RESUMO 
Investigou-se a existência de resíduos dos antimicrobianos enrofloxacina e ciprofloxacina em frangos 
expostos ao consumo em um município na região metropolitana de Curitiba, Brasil. Resíduos de 
antimicrobianos em tecidos animais acima dos limites permitidos (LMR) representam riscos à saúde 
humana. Em 10 amostras analisadas pelo laboratório Lanagro, do Ministério da Agricultura Pecuária 
e Abastecimento do Brasil, analisadas por Cromatografia Líquida Acoplada à Espectrometria de 
Massas, pela metodologia utilizada, não foram detectados resíduos dentro do limite mínimo de 20 
μg.kg-1 e máximo de 100 μg.kg-1 permitido. 
Palavras chave: Resíduos de antimicrobianos, tecidos de frangos, ciprofloxacina e enrofloxacina.
37 
INTRODUCTION 
Enrofloxacin is one of the most 
common antibiotics used in animal production 
(LOLO et al, 2006; REYES-HERRERA et al, 
2011) to control Escherichia coli and Salmonella 
(HARITOVA et al, 2011) infection. The main 
metabolite of this antimicrobial is ciprofloxacin 
(GARCIA et al, 2001; GARCIA et al, 2005), 
exclusively prescribed for treating severe 
infections in humans (BALL, 2000), especially 
those caused by β-lactam antibiotic-resistant 
bacteria (RUIZ et al, 2012). 
Antibiotic residues in products of 
animal origin are observed when veterinary 
practices are not followed, for example, when 
administering higher dosages than those 
prescribed, using products for longer than the 
recommended duration, and using a non-indicated 
route of administration (MICHELL et 
al, 1998). 
When present in food for human 
consumption, antibiotic residues in 
concentrations beyond the allowable limits may 
present some toxicological effects such as 
allergies, in addition to contributing to the 
selection of resistant bacteria in human gut 
microbiota, thereby compromising antibiotic 
therapies (LINDER et al, 2012; KIM et al, 2012). 
The maximum residue levels in foods 
allowed in Brazil are in accordance with Codex 
Alimentarius (Food and Agriculture 
Organization) and are authorized by the 
European Community. The monitoring control 
policies on food residues are established by the 
Normative Instruction DAS/MAA 42/1999 of 
the Ministry of Agriculture, Livestock and 
Supply and ratified by Normative Instruction no. 
7 of March 27, 2013. 
The Ministry of Agriculture, Livestock 
and Supply, through the laboratory Lanagro, is 
responsible for conducting liquid 
chromatographic analyses of chicken tissue 
samples collected by federal inspection bodies to 
verify the presence of quinolone residues. 
Publications reporting research results 
on antibiotic residues in chickens distributed for 
human consumption are either lacking or limited. 
METHODOLOGY 
In December 2011, a survey of major 
chicken brands sold for human consumption was 
conducted in Fazenda Rio Grande, in the 
metropolitan region of Curitiba. Seventeen 
brands were identified in supermarkets and meat 
markets. To sample each of the 10 predominant 
brands, namely Jaguá, Canção, Maringá, 
Goldfrango, Diplomata, Bigfrango, Aurora, 
Anhembi, Sadia, and Cooperave, one frozen 
chicken was purchased and 100 grams of its 
breast was removed. The samples were frozen at 
−20ºC and sent to the laboratory of the Ministry 
of Agriculture, Livestock and Supply (Lanagro), 
in the city of Porto Alegre, for liquid 
chromatographic analysis coupled with mass 
spectrometry. The analysis followed the internal 
protocol (method MET RPM/07/02), with 
minimum and maximum residue quantification 
levels of 20 and 100 μg/kg, respectively, as 
adopted by the National Plan for Control of 
Residues and Contaminants. The methodology 
used by Lanagro follows Directive 2002/657/EC 
of the European Union, which establishes 
performance indications, criteria, and procedures 
for analytical methods. 
RESULTS AND DISCUSSION 
The method used by Lanagro did not 
detect, in the 10 samples tested, any residues with 
concentrations within the allowable levels. 
A study on chickens distributed for 
human consumption in supermarkets and meat 
markets in the state capital of Brazil, conducted 
in 2011 by the Office of Consumer Protection in 
São Paulo, also did not detect any antibiotic 
residues, including enrofloxacin and 
ciprofloxacin, with concentrations within the 
allowable limits (IDEC, 2011). 
CONCLUSION 
Publications regarding the presence of 
antibiotic residues in animal tissues for human 
consumption in Brazil are lacking. 
At least for the chicken tissue samples 
and antibiotics assessed in this study, we can 
confirm that regulations regarding toxicological 
aspects in terms of allowable minimum and
38 
maximum residue levels in meat for domestic 
consumption are followed by agribusinesses in 
Brazil. 
BIBLIOGRAPHIC REFERENCES 
BAILAC S.; BARRÓN D.; SANZ-NEBOT V.; 
BARBOSA J. Determination of 
fluoroquinolones in chicken tissues by LC-coupled 
electrospray ionisation and atmospheric 
pressure chemical ionisation. J. Sep. Sci. v.29, 
n.1, p. 131 – 136, 2006. 
BALL, P. Quinolone generations: natural history 
or natural selection? Journal of Antimicrobial 
Chemotherapy. v.46, n.1, p.17-24, 2000. 
GARCIA, M. A; SOLANS, C.; HERNANDEZ, 
E.; PUIG, M.; BREGARTE, M. A. 
Simultaneous determination of enrofloxacin and 
its primary metabolite, ciprofloxacin in chicken 
tissues. Cromatographia. v.54, n.3/4, p.191- 
194, 2001. 
GARCIA, M.A.; SOLANS, C.; CALVO, A.; 
HERNANDEZ, E.; REY, R.; BREGANTE, 
M.A.; PUIG, M. Determination of Enrofloxacin 
and its primary metabolite, Ciprofloxacin, in pig 
tissues. Application to residue studies. 
Biomedical Chromatography, v.19, n.1, p.27- 
31, 2005. 
HARITOVA, A.; URUMOVA, V.; 
LUTCKANOV, M; PETROV, V.; LASHEV, L. 
Pharmacokinetic-pharmacodynamic indices of 
enrofloxacin in Escherichia coli O78/H12 
infected chickens. Food and Chemical 
Toxicology. v.49, p.1530–1536, 2011. 
INSTITUTO BRASILEIRO DE DEFESA DO 
CONSUMIDOR-IDEC. Produto Seguro, 
produção obscura. Revista 151. Fev.2011 
Disponível l em: http://www.idec.org.br/em-acao/ 
revista/151/materia/produto-seguro-produco- 
obscura Acesso em: 15/02/2012, 2011. 
KIM, B-S.; KIM, J. N.; YOON, S-H; CHUN, J; 
CERNIGLIA, C.E. Impact of enrofloxacin on the 
human intestinal microbiota revealed by 
comparative molecular analysis. Anaerobe. 
V.18, n.18, 310-320, 2012. 
LINDER, S W.; VEACH, B.T.; YAN, S.S, A; 
FERNÁNDEZ, H.; PINEIRO, S. A., AHN Y.; 
CERNIGLIA, C. E. In vitro enrofloxacin binding 
in human fecal slurries. Regulatory Toxicology 
and Pharmacology. v.62, n.1, 74–84. 2012. 
LOLO, M.; PEDREIRA, S.; MIRANDA, J. M.; 
VA´ZQUEZ, B. I.; FRANCO, C. M.; CEPEDA, 
A.; FENTE, C. Effect of cooking on enrofloxacin 
residues in chicken tissue. Food Additives and 
Contaminants. v.23, n.10, 988–993. 2006. 
MITCHELL, J.M.; GRIFFITHS, M.W.; 
MCEWENS, S.A; MCNAB, W.B; IEE L. 
Antimicrobial drug residues in milk and meat 
causeas, concerns prevalence, regulations, testx 
and test performance. Journal of food 
protection. v.61, n.6, p.742-756, 1998. 
REYES-HERRERA, I.; SCHNEIDER, M.J.; 
BLORE, P.J.; DONOGHUE, D. J. The 
relationship between blood and muscle samples 
to monitor for residues of the antibiotic 
enrofloxacin in chickens. Poultry Science. v.90, 
n.2, p. 481–485, 2011. 
RUIZ, J.; PONS, M.J; GOMES, C. Transferable 
mechanisms of quinolone resistance. 
International Journal of Antimicrobial 
Agents. v.40, n. 3, p. 196-203, 2012. 
_____________________________________ 
1-Nursing Department of the Santa Catarina 
State University (UDESC), Rua Licério Sperry, 
233 D, Bairro Engenho Braun, CEP 89809360, 
Chapecó, SC, Brazil. PhD student in 
Environment and Development. CAPES fellow. 
arkorb@yahoo.com.br 
2-Department of Collective Health of the Federal 
University of Paraná (UFPR) 
3-Pharmacology Department of the Federal 
University of Paraná (UFPR) 
4-Toxicology Department of the Federal 
University of Paraná (UFPR) 
5-Faculty of Veterinary Medicine, University of 
Lisbon (ULisboa)

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Artigo bioterra v14_n2_05

  • 1. REVISTA DE BIOLOGIA E CIÊNCIAS DA TERRA ISSN 1519-5228 36 Volume 14 - Número 2 - 2º Semestre 2014 RESIDUES OF ENROFLOXACIN AND CIPROFLOXACIN IN POULTRY TISSUES Arnildo Korb1; Eleusis R. de Nazareno2; Paulo R. Dalsenter3; Ciro A. de O. Ribeiro4; Constança Pomba5 ABSTRACT We investigated the existence of residues of the antimicrobials enrofloxacin and ciprofloxacin in broilers, which are exposed to the consumer in the metropolitan city Curitiba, Brazil. Antimicrobial residues in animal tissues, above the minimum allowed concentration, pose risks to human health. In 10 samples analyzed by Liquid Chromatography (Mass Spectometry), at the Lanagro laboratory, from Ministry of Agriculture, Livestock and Supply of Brazil, considering a lower limit of 20 μg.kg-1 and maximum 100 μg.kg-1, no residues of ciprofloxacin and enrofloxacin were detected. These results showed that in the broiler samples of different brands, these antimicrobials were not detected. Keywords: Antimicrobial residues, poultry tissues, ciprofloxacin and enrofloxacin. RESÍDUOS DE ENROFLOXACINA E CIPROFLOXACINA EM TECIDOS DE FRANGOS RESUMO Investigou-se a existência de resíduos dos antimicrobianos enrofloxacina e ciprofloxacina em frangos expostos ao consumo em um município na região metropolitana de Curitiba, Brasil. Resíduos de antimicrobianos em tecidos animais acima dos limites permitidos (LMR) representam riscos à saúde humana. Em 10 amostras analisadas pelo laboratório Lanagro, do Ministério da Agricultura Pecuária e Abastecimento do Brasil, analisadas por Cromatografia Líquida Acoplada à Espectrometria de Massas, pela metodologia utilizada, não foram detectados resíduos dentro do limite mínimo de 20 μg.kg-1 e máximo de 100 μg.kg-1 permitido. Palavras chave: Resíduos de antimicrobianos, tecidos de frangos, ciprofloxacina e enrofloxacina.
  • 2. 37 INTRODUCTION Enrofloxacin is one of the most common antibiotics used in animal production (LOLO et al, 2006; REYES-HERRERA et al, 2011) to control Escherichia coli and Salmonella (HARITOVA et al, 2011) infection. The main metabolite of this antimicrobial is ciprofloxacin (GARCIA et al, 2001; GARCIA et al, 2005), exclusively prescribed for treating severe infections in humans (BALL, 2000), especially those caused by β-lactam antibiotic-resistant bacteria (RUIZ et al, 2012). Antibiotic residues in products of animal origin are observed when veterinary practices are not followed, for example, when administering higher dosages than those prescribed, using products for longer than the recommended duration, and using a non-indicated route of administration (MICHELL et al, 1998). When present in food for human consumption, antibiotic residues in concentrations beyond the allowable limits may present some toxicological effects such as allergies, in addition to contributing to the selection of resistant bacteria in human gut microbiota, thereby compromising antibiotic therapies (LINDER et al, 2012; KIM et al, 2012). The maximum residue levels in foods allowed in Brazil are in accordance with Codex Alimentarius (Food and Agriculture Organization) and are authorized by the European Community. The monitoring control policies on food residues are established by the Normative Instruction DAS/MAA 42/1999 of the Ministry of Agriculture, Livestock and Supply and ratified by Normative Instruction no. 7 of March 27, 2013. The Ministry of Agriculture, Livestock and Supply, through the laboratory Lanagro, is responsible for conducting liquid chromatographic analyses of chicken tissue samples collected by federal inspection bodies to verify the presence of quinolone residues. Publications reporting research results on antibiotic residues in chickens distributed for human consumption are either lacking or limited. METHODOLOGY In December 2011, a survey of major chicken brands sold for human consumption was conducted in Fazenda Rio Grande, in the metropolitan region of Curitiba. Seventeen brands were identified in supermarkets and meat markets. To sample each of the 10 predominant brands, namely Jaguá, Canção, Maringá, Goldfrango, Diplomata, Bigfrango, Aurora, Anhembi, Sadia, and Cooperave, one frozen chicken was purchased and 100 grams of its breast was removed. The samples were frozen at −20ºC and sent to the laboratory of the Ministry of Agriculture, Livestock and Supply (Lanagro), in the city of Porto Alegre, for liquid chromatographic analysis coupled with mass spectrometry. The analysis followed the internal protocol (method MET RPM/07/02), with minimum and maximum residue quantification levels of 20 and 100 μg/kg, respectively, as adopted by the National Plan for Control of Residues and Contaminants. The methodology used by Lanagro follows Directive 2002/657/EC of the European Union, which establishes performance indications, criteria, and procedures for analytical methods. RESULTS AND DISCUSSION The method used by Lanagro did not detect, in the 10 samples tested, any residues with concentrations within the allowable levels. A study on chickens distributed for human consumption in supermarkets and meat markets in the state capital of Brazil, conducted in 2011 by the Office of Consumer Protection in São Paulo, also did not detect any antibiotic residues, including enrofloxacin and ciprofloxacin, with concentrations within the allowable limits (IDEC, 2011). CONCLUSION Publications regarding the presence of antibiotic residues in animal tissues for human consumption in Brazil are lacking. At least for the chicken tissue samples and antibiotics assessed in this study, we can confirm that regulations regarding toxicological aspects in terms of allowable minimum and
  • 3. 38 maximum residue levels in meat for domestic consumption are followed by agribusinesses in Brazil. BIBLIOGRAPHIC REFERENCES BAILAC S.; BARRÓN D.; SANZ-NEBOT V.; BARBOSA J. Determination of fluoroquinolones in chicken tissues by LC-coupled electrospray ionisation and atmospheric pressure chemical ionisation. J. Sep. Sci. v.29, n.1, p. 131 – 136, 2006. BALL, P. Quinolone generations: natural history or natural selection? Journal of Antimicrobial Chemotherapy. v.46, n.1, p.17-24, 2000. GARCIA, M. A; SOLANS, C.; HERNANDEZ, E.; PUIG, M.; BREGARTE, M. A. Simultaneous determination of enrofloxacin and its primary metabolite, ciprofloxacin in chicken tissues. Cromatographia. v.54, n.3/4, p.191- 194, 2001. GARCIA, M.A.; SOLANS, C.; CALVO, A.; HERNANDEZ, E.; REY, R.; BREGANTE, M.A.; PUIG, M. Determination of Enrofloxacin and its primary metabolite, Ciprofloxacin, in pig tissues. Application to residue studies. Biomedical Chromatography, v.19, n.1, p.27- 31, 2005. HARITOVA, A.; URUMOVA, V.; LUTCKANOV, M; PETROV, V.; LASHEV, L. Pharmacokinetic-pharmacodynamic indices of enrofloxacin in Escherichia coli O78/H12 infected chickens. Food and Chemical Toxicology. v.49, p.1530–1536, 2011. INSTITUTO BRASILEIRO DE DEFESA DO CONSUMIDOR-IDEC. Produto Seguro, produção obscura. Revista 151. Fev.2011 Disponível l em: http://www.idec.org.br/em-acao/ revista/151/materia/produto-seguro-produco- obscura Acesso em: 15/02/2012, 2011. KIM, B-S.; KIM, J. N.; YOON, S-H; CHUN, J; CERNIGLIA, C.E. Impact of enrofloxacin on the human intestinal microbiota revealed by comparative molecular analysis. Anaerobe. V.18, n.18, 310-320, 2012. LINDER, S W.; VEACH, B.T.; YAN, S.S, A; FERNÁNDEZ, H.; PINEIRO, S. A., AHN Y.; CERNIGLIA, C. E. In vitro enrofloxacin binding in human fecal slurries. Regulatory Toxicology and Pharmacology. v.62, n.1, 74–84. 2012. LOLO, M.; PEDREIRA, S.; MIRANDA, J. M.; VA´ZQUEZ, B. I.; FRANCO, C. M.; CEPEDA, A.; FENTE, C. Effect of cooking on enrofloxacin residues in chicken tissue. Food Additives and Contaminants. v.23, n.10, 988–993. 2006. MITCHELL, J.M.; GRIFFITHS, M.W.; MCEWENS, S.A; MCNAB, W.B; IEE L. Antimicrobial drug residues in milk and meat causeas, concerns prevalence, regulations, testx and test performance. Journal of food protection. v.61, n.6, p.742-756, 1998. REYES-HERRERA, I.; SCHNEIDER, M.J.; BLORE, P.J.; DONOGHUE, D. J. The relationship between blood and muscle samples to monitor for residues of the antibiotic enrofloxacin in chickens. Poultry Science. v.90, n.2, p. 481–485, 2011. RUIZ, J.; PONS, M.J; GOMES, C. Transferable mechanisms of quinolone resistance. International Journal of Antimicrobial Agents. v.40, n. 3, p. 196-203, 2012. _____________________________________ 1-Nursing Department of the Santa Catarina State University (UDESC), Rua Licério Sperry, 233 D, Bairro Engenho Braun, CEP 89809360, Chapecó, SC, Brazil. PhD student in Environment and Development. CAPES fellow. arkorb@yahoo.com.br 2-Department of Collective Health of the Federal University of Paraná (UFPR) 3-Pharmacology Department of the Federal University of Paraná (UFPR) 4-Toxicology Department of the Federal University of Paraná (UFPR) 5-Faculty of Veterinary Medicine, University of Lisbon (ULisboa)