Home  |  About JAPTR |  Editorial board  |  Search |  Ahead of print  |  Current issue  |  Archives |  Submit article  |  Instructions  |  Subscribe  |  Advertise  |  Contacts  |Login 
Users Online: 1139   Home Print this page Email this page Small font sizeDefault font sizeIncrease font size
     

 Table of Contents  
ORIGINAL ARTICLE
Year : 2016  |  Volume : 7  |  Issue : 1  |  Page : 2-5  

Quantification of residual enrofloxacin and ciprofloxacin in feathers of broiler chickens by high-performance liquid chromatography-fluorescence after oral administration of the drugs


Department of Pharmacology, National University of La Plata, Conicet, La Plata, Argentina

Date of Web Publication6-Jan-2016

Correspondence Address:
Gustavo H Marin
Department of Pharmacology, National University of La Plata, La Plata
Argentina
Login to access the Email id

Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2231-4040.173265

Rights and Permissions
  Abstract 

Enrofloxacin (ENR) and ciprofloxacin (CIP) are drugs used in poultry feeding. In general, feathers that are incorporated in the food chain as a protein source for animal feed, have residues of these drugs. In order to study the pharmacokinetic of ENR/CIP residues in feathers of broiler chickens, to calculate the waiting times for these drugs, before human consumption, we developed the present research. Feathers of broiler chickens were enriched with ENR/CIP. After adding acetone, the mix was agitated and centrifuged and supernatant evaporated under nitrogen. The dry residue was suspended in a tetrahydrofuran solution and the supernatant was injected into the chromatographic system for analysis. Animals showed high levels of ENR/CIP in their feathers after administration of 10 mg/kg enrofloxacin dissolved in drinking water for 5 days. Both compounds were detected in feathers during 9 days. The analytical method developed in this paper to determine ENR and CIP in feathers of broiler chicken showed good linearity, selectivity, accuracy and precision in the analysis conditions. This technique could have important applications in the studies on residues of ENR/CIP in feathers, since the effect of this component in animal diets has not been considered yet.

Keywords: Enrofloxacin, quantification, quinolones


How to cite this article:
Haag G, Marin GH, Errecalde J. Quantification of residual enrofloxacin and ciprofloxacin in feathers of broiler chickens by high-performance liquid chromatography-fluorescence after oral administration of the drugs. J Adv Pharm Technol Res 2016;7:2-5

How to cite this URL:
Haag G, Marin GH, Errecalde J. Quantification of residual enrofloxacin and ciprofloxacin in feathers of broiler chickens by high-performance liquid chromatography-fluorescence after oral administration of the drugs. J Adv Pharm Technol Res [serial online] 2016 [cited 2021 Sep 16];7:2-5. Available from: https://www.japtr.org/text.asp?2016/7/1/2/173265


  Introduction Top


Antibiotics used in animal feed have become a matter of major public health considering that their residues might be dangerous for both animals and humans due to toxicological effects and development of resistances.[1]

Enrofloxacin (ENR), a drug of the quinolones group, is an antimicrobial commonly used in the treatment and prevention of poultry diseases.

Feathers are not used as a matrix for detection and quantification of veterinary medication. However, they are frequently incorporated as a protein source in diets of other animals, such as cattle, swine, trout, and salmons.

Kinetic studies in edible tissues of poultry allow determining the waiting time of ENR and its metabolite ciprofloxacin (CIP) to ensure safe foods for human consumption. In general, the presence of residues in feathers that are incorporated in the food chain as a protein source in animal feed are not controlled and waiting times for ENR and CIP are not determined.

Since the human nutrition incorporates this animal's meat to the diet and considering that the antibiotics could be present in that meat, we initiated the present study with the following goals:

  • To develop a method to quantify residues of ENR and its metabolite CIP in feathers of broiler chickens using high-performance liquid chromatography-fluorescence; this method has been specifically validated for feathers
  • To study the pharmacokinetic of ENR and CIP residues in feathers of broiler chickens.



  Materials and Methods Top


The extraction procedure was a modification of the technique developed by San Martín et al.[2] Feathers of broiler chickens were homogenized and enriched with ENR and CIP. After adding acetone, the mix was vigorously agitated and centrifuged. The supernatant obtained was evaporated under nitrogen. The dry residue was resuspended in a tetrahydrofuran solution, and after a second centrifugation, the supernatant was injected into the chromatographic system [Figure 1].
Figure 1: Extraction procedure ENR: Enrofloxacin, CIP: Ciprofloxacin

Click here to view


Chromatography conditions are mobile phase (water, acetonitrile, triethylamine), flow rate at 1,2 ml/min and column C18 (Phenomenex Luna).[3]

The validation procedure was performed following Commission Decision 2002/657/EC of the UE.[4]

Validation parameters included linearity, selectivity, specificity, limit of detection, limit of quantification, accuracy, and precision intra- and inter-day.

Three-week-old broiler chickens were given 10 mg of ENR/kg of body weight for consecutive 5 days. ENR was dissolved in drinking water.

Samples of feathers from 10 chickens were randomly taken on the day 0, 1, 2, 3, 4, 5, 7, and 9, after the last administration of the antimicrobial. The feathers were prepared, labeled, and stored until extraction and chromatographic assay. The extraction procedure and chromatography conditions were described in another summary submitted in the present event.

Calibration standards

The calibration curve was linear in the range of concentrations assessed with a correlation coefficient of 0.9974 for ENR and 0.9986 for CIP. The limits of detection were 0.062 µg/g and 0.040 µg/g and the quantification limits were 0.080 µg/g and 0.050 µg/g for ENR and CIP, respectively [Figure 2] and [Figure 3].
Figure 2: Calibration curve of standard enrofloxacin

Click here to view
Figure 3: Calibration curve of standard ciprofloxacin

Click here to view


The specificity of the method was demonstrated by the analysis of blank samples when compared with standards of ENR and CIP.

Inter- and intra-day accuracy expressed by the variation coefficient was 3.88 at 7.47 in ENR and 7.07 at 2.73 in CIP. Recoveries were 94.57 at 100% and 93.33 at 100% for ENR and its metabolite CIP, respectively [Table 1] and [Table 2].
Table 1: Recovery of enrofloxacin in chicken feathers enriched with 0.1, 0.25, and 2 µg/g

Click here to view
Table 2: Recovery of ciprofloxacin in chicken feathers enriched with 0.1, 0.25, and 2 µg/g

Click here to view



  Results Top


A group of 10 experimental animals were included in the experience. These specimens showed high levels of ENR and CIP in their feathers after administration of 10 mg/kg ENR dissolved in drinking water for 5 days.

Although the antibiotic residue was similar by the end of the 1st day, CIP has shown slightly higher levels than ENR at the day 3 and 5 after inoculation.

Both compounds were detected in feathers, during 9 days [Figure 4]. After the 10th day, it could be said that antibiotic residue and that after this day samples were clean of drugs residues and suitable for consumption.
Figure 4: Blood level of drugs according to time administration

Click here to view



  Discussion Top


Feather flour is a potential source of drug residues that can reach humans through the food chain when it is incorporated as a component in the diet of animals for human consumption. The feathers analyzed in the present study showed a high concentration of ENR and CIP. This finding cannot be explained by blood contribution to this tissue since feather vasculature reaches only the lower portion of the calamus. Being penetrating substances with high solubility, it is possible to hypothesize that a source of feather contamination could be the secretion from the uropygial gland that reaches the feathers via grooming behavior.[5]

Therefore, it becomes an extremely important decision to identify antibiotic residues in food destination to human consumption, and also to determinate the removal time period for use as food feathers of other animals, in order to avoid antibiotic resistance in feed chain.

On the other hand, this study exposes a new and effective technique to analyze still not so well-studied tissue in order to give new options for future antibiotics research in birds.

In addition, analyzing withdrawal time for slaughter according to different trademarks, the period would be between 8 and 10 days and this time will be enough to eliminate more than 95% in birds' feathers.


  Conclusions Top


The novelty of the method is based on the possibility of a real and effective dosage form in the feathers. Besides, it opens the debate on the awareness of the possibility of antibiotic resistance if the time of withdrawal of the antibiotic is not respected even if that food is not destined for human consumption, since it becomes a part of the animal feed chain.

Therefore, the analytical method developed to determine ENR and CIP in pens of broilers showed good linearity, selectivity, accuracy, and precision.

This technique could have important applications in the studies on residues of ENR and CIP in feathers since the effect of this component in animal diets has not been considered yet.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

 
  References Top

1.
Sapkota AR, Lefferts L, McKenzie S, Walker P. What do we feed to food production animals? A review of animal feed ingredients and their potential impacts on human health. Environ Health Perspect 2007. doi:10.1289/ehp.9760  Back to cited text no. 1
    
2.
San Martín B, Cornejo J, Iragüen D, Hidalgo H, Anadón A. Depletion study of enrofloxacin and its metabolite ciprofloxacin in edible tissues and feathers of white leghorn hens by liquid chromatography coupled with tandem mass spectrometry. J Food Prot 2007;70:1952-7.  Back to cited text no. 2
    
3.
Mestorino N. Kinetic of Enrofloxacin XVIII Argentine Conference and XIII Latin American Conference on Drug-Veterinary Toxicology. Vol. 2. FCV, UBA; 2008. p. 93-4.  Back to cited text no. 3
    
4.
European Union. Commission Decision 2002/657/EC implementing Council Directive 96/23/EC on the application of analytical methods and the interpretation of results. Community J Eur 2002;221:23-33.  Back to cited text no. 4
    
5.
López-Cadenas C, Sierra-Vega M, García-Vieitez JJ, Diez-Liébana MJ, Sahagún-Prieto A, Fernández-Martínez N. Enrofloxacin: Pharmacokinetics and metabolism in domestic animal species. Curr Drug Metab 2013;14:1042-58.  Back to cited text no. 5
    


    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4]
 
 
    Tables

  [Table 1], [Table 2]


This article has been cited by
1 Tetracyclines in Processed Animal Proteins: A Monitoring Study on Their Occurrence and Antimicrobial Activity
Sara Morello,Sabina Pederiva,Rosa Avolio,Giuseppina Amato,Simona Zoppi,Alessia Di Blasio,Maria Cesarina Abete,Cristina Casalone,Rosanna Desiato,Giuseppe Ru,Daniela Marchis
Foods. 2021; 10(4): 696
[Pubmed] | [DOI]
2 Impact of antibiotic concentration gradients on nitrate reduction and antibiotic resistance in a microfluidic gradient chamber
Lang Zhou,Reinaldo E. Alcalde,Jinzi Deng,Baltazar Zuniga,Robert A. Sanford,Bruce W. Fouke,Charles J. Werth
Science of The Total Environment. 2021; 779: 146503
[Pubmed] | [DOI]
3 Residue study of nitroimidazoles depletion in chicken feathers in comparison with some other selected matrixes
Ales Církva,Irena Málková,Martina Rejtharová,Eva Vernerová,Alfred Hera,Jirí Bureš
Food Additives & Contaminants: Part A. 2019; : 1
[Pubmed] | [DOI]
4 The Use of Feathers from Racing Pigeons for Doping Control Purposes
Fernando X Moreira,Helena Carmo,Armindo Melo,Maria B André,Renata Silva,Zélia Azevedo,Maria L Bastos,Paula G de Pinho
Journal of Analytical Toxicology. 2018;
[Pubmed] | [DOI]
5 Presence of residues and metabolites of pharmaceuticals in environmental compartments, food commodities and workplaces: A review spanning the three-year period 2014–2016
Paola Bottoni,Sergio Caroli
Microchemical Journal. 2017;
[Pubmed] | [DOI]
6 Clinical Efficacy and Residue Depletion of 10% Enrofloxacin Enteric-Coated Granules in Pigs
Zhixin Lei,Qianying Liu,Bing Yang,Jincheng Xiong,Kun Li,Saeed Ahmed,Liping Hong,Pin Chen,Qigai He,Jiyue Cao
Frontiers in Pharmacology. 2017; 8
[Pubmed] | [DOI]
7 A NIR-responsive up-conversion nanoparticle probe of the NaYF4:Er,Yb type and coated with a molecularly imprinted polymer for fluorometric determination of enrofloxacin
Yiwei Tang,Min Li,Xue Gao,Xiuying Liu,Jinwen Gao,Tao Ma,Jianrong Li
Microchimica Acta. 2017;
[Pubmed] | [DOI]



 

Top
 
 
  Search
 
    Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
    Access Statistics
    Email Alert *
    Add to My List *
* Registration required (free)  

 
  In this article
   Abstract
  Introduction
   Materials and Me...
  Results
  Discussion
  Conclusions
   References
   Article Figures
   Article Tables

 Article Access Statistics
    Viewed4112    
    Printed56    
    Emailed0    
    PDF Downloaded446    
    Comments [Add]    
    Cited by others 7    

Recommend this journal