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A HPTLC-Fluorescent Densitometry Assay for Simultaneous Detection of Enrofloxacin and Ciprofloxacin in Broiler Chicken Tissues

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Abstract

A high-performance thin-layer chromatographic (HPTLC) method was developed and validated according to the protocol on Validation of Analytical Procedures: methodology, Veterinary International Cooperation on Harmonization for simultaneous determination of enrofloxacin and ciprofloxacin in broiler chicken tissues viz. liver, kidney, muscle-breast, muscle-thigh, and skin. Chromatography was performed on thin-layer chromatography (TLC) silica gel 60F254, aluminum sheets (10 × 10 cm) by Camag Linomat-5 applicator. The isolation of enrofloxacin and ciprofloxacin was attained by two development stages, initially with diethyl ether followed by dichloromethane/methanol/aqueous ammonia/acetonitrile (2:3:2:3) as mobile phase. The developed TLC plates were exposed to hydrochloric acid fumes and fluorescent densitometric evaluation was performed at 366 nm using Camag TLC Scanner-3 with WinCAT 1.4.4 software. The RF value was 0.60 and 0.44 for enrofloxacin and ciprofloxacin, respectively. The detection limits were 2 and 3 ng/band for enrofloxacin and ciprofloxacin, respectively, and 5 ng/band as limit of quantification for both the compounds. Enrofloxacin and ciprofloxacin showed wide linear range from 2 to 110 ng/band and 3 to 110 ng/band with high correlation coefficient values of 0.99997 and 0.99990 for peak area, respectively. An exemplary precision was observed for individual compounds. The percent recovery for ciprofloxacin was 82.0–86.8% when compared to enrofloxacin 83.4–90.3%. The observed results were within the acceptable values as a function of the analyte concentration, suggested by the Association of Official Analytical Chemists peer-verified methods program. The present validated HPTLC method can be used as screening technique for antibiotics residue monitoring program in broiler chicken tissues to promulgate food safety.

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Acknowledgements

The Authors are highly thankful to the Drugs and Pharmaceutical Research Programme, Department of Science and Technology (DST), Government of India, New Delhi, for the financial assistance in conducting the experiment as part of the DST scheme entitled “A National Facility for Pharmacovigilance on Drug Residue and other Toxic Xenobiotics including Genetically Manipulated Organisms in Veterinary Products” at Pharmacovigilance Laboratory for Animal Feed and Food Safety, Directorate of Centre for Animal Health Studies, Tamil Nadu Veterinary and Animal Sciences University, Chennai-600 051.

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  1. Ghadevaru Sarathchandra

    Present address: Pharmacovigilance Laboratory for Animal Feed and Food Safety, Directorate of Centre for Animal Health Studies, Tamil Nadu Veterinary and Animal Sciences University, Madhavaram Milk Colony, Chennai, 600 051, India

Authors and Affiliations

  1. Department of Veterinary Pharmacology and Toxicology, Veterinary College and Research Institute, Tirunelveli, Tamil Nadu, 627 358, India

    Veerapandian Sureshkumar

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  1. Veerapandian Sureshkumar
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  2. Ghadevaru Sarathchandra
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Correspondence to Ghadevaru Sarathchandra.

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Veerapandian Sureshkumar declares that he has no conflict of interest.

Ghadevaru Sarathchandra declares that he has no conflict of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Part of Ph.D. thesis submitted to Tamil Nadu Veterinary and Animal Sciences University, Tamil Nadu State, India.

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Sureshkumar, V., Sarathchandra, G. A HPTLC-Fluorescent Densitometry Assay for Simultaneous Detection of Enrofloxacin and Ciprofloxacin in Broiler Chicken Tissues. Food Anal. Methods 11, 1076–1085 (2018). https://doi.org/10.1007/s12161-017-1077-x

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  • DOI: https://doi.org/10.1007/s12161-017-1077-x

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