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摘要
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 60F(254), aluminum sheets (10 x 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 R-F 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.
- 出版日期2018-4
