ABSTRACT
Immunoassays greatly contribute to veterinary drug residue analysis. However, there are few reports on detecting neomycin residues by immunoassay. Here, a rapid and sensitive chemiluminescent enzyme immunoassay (CLIEA) was successfully developed for neomycin residue analysis. CLIEA demonstrated good cross-reactivity for neomycin, and the IC50 value was 2.4 ng/mL in buffer. The average recovery range was 88.5%-105.4% for spiked samples (10, 50, and 100 μg/kg), and the coefficient of variation was in the range of 7.5%-14.5%. The limit of detection of CLEIA was 9.4 μg/kg, and this method was compared with the liquid chromatography-tandem mass spectrometry method using naturally contaminated samples, producing a correlation coefficient of >0.95. We demonstrate a reliable CLIEA for the rapid screening of neomycin in milk.
Subject(s)
Animals , Anti-Bacterial Agents , Metabolism , Drug Residues , Metabolism , Food Contamination , Immunoenzyme Techniques , Limit of Detection , Luminescent Measurements , Milk , Chemistry , Neomycin , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To determine 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) residues released from protein bound AMOZ in animal tissues.</p><p><b>METHODS</b>Polyclonal and monoclonal antibodies were produced in this study. A rapid, sensitive, and specific competitive direct enzyme-linked immunosorbent assay (cdELISA) was developed.</p><p><b>RESULTS</b>Rabbit polyclonal antibodies were used in the optimized cdELISA method, and exhibited negligible cross-reactivity with other compounds structurally related to AMOZ. The IC(50) of the polyclonal antibody was 0.16 ng/mL. The method limit of detection in four different types of animal and fish tissues was less than 0.06 μg/kg. Recoveries ranged from 80% to 120% for fortified samples with the coefficient of variation values less than 15%. The results of the cdELISA method were in good agreement with the results from an established liquid chromatography-tandem mass spectrometry confirmatory method used for AMOZ residues.</p><p><b>CONCLUSION</b>The cdELISA method developed in the present study is a convenient practical tool for screening large numbers of animal and fish tissue samples for the the detection of released protein bound AMOZ residues.</p>
