A HPTLC-Fluorescent Densitometry Assay for Simultaneous Detection of Enrofloxacin and Ciprofloxacin in Broiler Chicken T
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A HPTLC-Fluorescent Densitometry Assay for Simultaneous Detection of Enrofloxacin and Ciprofloxacin in Broiler Chicken Tissues Veerapandian Sureshkumar 1 & Ghadevaru Sarathchandra 2
Received: 20 June 2017 / Accepted: 23 October 2017 # Springer Science+Business Media, LLC 2017
Abstract A high-performance thin-layer chromatographic (HPTLC) method was developed and validated according to the protocol on Validation of Analytical Procedures: methodo l o g y, Ve t e r i n a r y I n t e r n a t i o n a l C o o p e r a t i o n o n 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
Part of Ph.D. thesis submitted to Tamil Nadu Veterinary and Animal Sciences University, Tamil Nadu State, India. * Ghadevaru Sarathchandra [email protected] Veerapandian Sureshkumar [email protected] 1
Department of Veterinary Pharmacology and Toxicology, Veterinary College and Research Institute, Tirunelveli, Tamil Nadu 627 358, India
2
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
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. Keywords HPTLC . Enrofloxacin . Ciprofloxacin . Broiler chicken tissues
Introduction The widespread use of fluoroquinolones as therapeutic and prophylactic agents, in food animals particularly in intensive poultry production, has become a matter of considerable concern in recent years due to the ident
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