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Title: Development of an enzyme-linked immunosorbent assay for determination of the furaltadone etabolite, 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) in animal tissues

Author
item LUO, PENG - Chinese Center For Disease Control
item JIANG, WEN - China Agricultural University
item Beier, Ross
item SHEN, JIAN - China Agricultural University
item JIANG, HAI - China Agricultural University
item MIAO, HONG - Chinese Center For Disease Control
item ZHAO, YUN - Chinese Center For Disease Control
item CHEN, XIA - Chinese Center For Disease Control
item WU, YONG - Chinese Center For Disease Control

Submitted to: Biomedical and Environmental Sciences
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/29/2012
Publication Date: 8/28/2012
Citation: Luo, P.J., Jiang, W.X., Beier, R.C., Shen, J.Z., Jiang, H.Y., Miao, H., Zhao, Y.F., Chen, X., Wu, Y.N. 2012. Development of an enzyme-linked immunosorbent assay for determination of the furaltadone metabolite, 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) in animal tissues. Biomedical and Environmental Sciences. 25:449-457.

Interpretive Summary: Furaltadone is a broad-spectrum, anti-bactericidal drug that belongs in the class of nitrofurans. However, the metabolite of furaltadone, 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ), is toxic to humans. There is currently interest in implementation of rapid screening methods for the detection of nitrofuran metabolite residues in animal tissues. We have developed antibodies that recognize AMOZ. Antibodies are substances that are produced by the immune system in response to foreign substances that enter the body. Once the antibodies to a foreign substance are isolated, they can be used in a method to detect the presence of that foreign substance. The antibodies that we isolated may be used in an easy-to-use test called an immunoassay (producing results similar to the pregnancy test) for the simple, rapid, and sensitive screening of AMOZ residues. The immunoassay was validated by submitting identical samples for analysis by mass spectrometry methods. The overall results indicate that the immunoassay method developed here is a convenient practical tool for screening large numbers of animal and fish tissue samples for the detection of the toxic AMOZ residues.

Technical Abstract: A rapid, sensitive, and specific competitive direct enzyme-linked immunosorbent assay (cdELISA) for determination of protein bound 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) residues is described to monitor the illegal use of furaltadone. Polyclonal and monoclonal antibodies were produced in this study. Rabbit polyclonal antibodies were used in the optimized cdELISA method and exhibited negligible cross-reactivity with other compounds structurally related to AMOZ. The IC50 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 cdELISA method was validated by a previously established liquid chromatography–tandem mass spectrometry method. These results indicate that the cdELISA method developed here is a convenient, practical tool for screening large numbers of animal and fish tissue samples for the detection of AMOZ residues.