Simultaneous Determination of Florfenicol and Its Metabolite Florfenicol Amine in Swine Muscle Tissue by a Heterologous Enzyme-Linked Immunosorbent Assay
Abstract:A sensitive and heterologous enzyme-linked immunosorbent assay (ELISA) for the simultaneous detection of florfenicol (FF) and its metabolite florfenicol amine (FFA) in swine muscle tissue was developed. FFA was conjugated to bovine serum albumin by a formaldehyde coupling method as an immunogen to immunize rabbits. FFA, thiamphenicol glycinate, and modified FF were conjugated to ovalbumin as coating antigens. The effect of different types of hapten heterology on the sensitivity and specificity of the ELISA was evaluated. Using FF glutaric anhydride ester as a coating hapten and antibody raised against modified FFA, an ELISA was developed that showed an IC50 value of 0.48 ng/mL. The antibody showed a cross-reactivity of 100 with FFA, 97 with FF, 6 with thiamphenicol, and a negligible value with chloramphenicol. From fortified swine muscle samples at levels of 4320 ng/g, the average recoveries of FF and FFA ranged from 58.2 to 96.8, with coefficients of variation less than 14. Analysis of incurred samples by the ELISA gave similar results to those by a previously developed liquid chromatographic method. The ELISA could be used as a rapid method for the simultaneous determination of FF and FFA in swine muscle tissue.
Document Type: Research Article
Affiliations: 1: China Agricultural University, College of Veterinary Medicine, Department of Veterinary Pharmacology and Toxicology, and China Agricultural University, National Reference Laboratory for Veterinary Drug Residue, Beijing 100193, People's Republic of China. 2: China Agricultural University, Beijing Wanger Biotechnology Co. Ltd, Beijing 100193, People's Republic of China.
Publication date: May 28, 2009
- The Journal of AOAC INTERNATIONAL publishes refereed papers and reviews in the fields of chemical, biological and toxicological analytical chemistry for the purpose of showcasing the most precise, accurate and sensitive methods for analysis of foods, food additives, supplements and contaminants, cosmetics, drugs, toxins, hazardous substances, pesticides, feeds, fertilizers and the environment available at that point in time. The scope of the Journal includes unpublished original research describing new analytical methods, techniques and applications; improved approaches to sampling, both in the field and the laboratory; better methods of preparing samples for analysis; collaborative studies substantiating the performance of a given method; statistical techniques for evaluating data. The Journal will also publish other articles of general interest to its audience, e.g., technical communications; cautionary notes; comments on techniques, apparatus, and reagents.
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