Purpose: Enzyme-linked immunosorbent assays (ELISAs) were developed to screen for florfenicol and fluoroquinolones in animal tissues. Performance of the ELISAs were validated according to 2002/657/EC guidelines and results reported.
Methods: Following 2002/657/EC guidelines for screening assays, validation for florfenicol screening was completed by collecting ELISA data on extracts from 60 different Salmonidae muscle samples spiked with 1,000 ppb florfenicol and 60 negative controls. Use for avian muscle was then validated by collecting ELISAs on extracts from 20 avian muscle samples spiked with 150 ppb florfenicol and 20 negative controls. Similarly, validation of the extraction and ELISA for fluoroquinolones was completed in shrimp and avian muscle.
Results: Mean ELISA results were 1,474 ± 473 ppb for 1,000 ppb florfenicol spiked Salmonidae muscle samples, 11 ± 19 ppb for unspiked, 199 ± 87 ppb for 150 ppb spiked avian muscle and 12 ± 12 ppb for unspiked. Three of 60 spiked Salmonidae and 1 of 20 of the spiked avian muscle samples were false compliant. Mean ELISA results were 0.90 ± 0.22 ppb for 1.0 ppb enrofloxacin spiked shrimp samples, 0.15 ± 0.09 ppb for unspiked, 1.16 ± 0.25 ppb for spiked avian muscle samples, and 0.50 ± 0.13 ppb for unspiked. One of 60 enrofloxacin spiked shrimp samples and 0 of 20 enrofloxacin spiked avian muscle samples were false compliant. All data sets conformed to 2002/657/EC criteria for screening methods.
Significance: To help ensure animal protein products are not contaminated with drug residues, validated ELISA screening methods are now available for detection of florfenicol and fluoroquinolones in animal tissues.