Development of a competitive ELISA for the detection of a furaltadone marker residue, 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ), in cultured fish samples.
This report describes an enzyme-linked immunosorbent assay (ELISA) for tissue-bound metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) and the application to residue analysis in cultured fish samples. The residue is monitored as a marker for the drug furaltadone. The assay enables the detection of protein bound AMOZ in the form of a 2-nitrophenyl derivative (2-NP-AMOZ) in sample supernatant or extract after acid hydrolysis and derivatization with o-nitrobenzaldehyde. Polyclonal rabbit antibodies were produced with a new immunogen hapten, 2-NP-HXA-AMOZ. The new ELISA had adequate analytical sensitivity (IC(50) value 0.325 μg kg(-1); limit of detection 0.1 μg kg(-1)) to determine a trace of AMOZ residue and had a high selectivity. Recoveries of AMOZ fortified at the levels of 0.1, 0.5 and 1.0 μg kg(-1) ranged from 89.8 to 112.5% with coefficients of variation of 12.4-16.2% over the range of AMOZ concentrations studied. The results obtained with the ELISA correlated well with those obtained by commercial test kits for 150 tested samples (r=0.984). The results suggest that the developed ELISA is a highly specific, accurate, and sensitive method suitable for high throughput screening for AMOZ residues.
School of Chinese Medicine, China Medical University, Taichung City 404, Taiwan, ROC.
Shi-Yuan
SY
Sheu
Yung-Te
YT
Tai
Wen-Ren
WR
Li
Yi-Chih
YC
Lei
Kuan-Huei
KH
Hsieh
Chiu-Yueh
CY
Lin
Chu-Chen
CC
Cheng
Tong-Hsuan
TH
Chang
Tzong-Fu
TF
Kuo
D000818:Animals; D001547:Benzaldehydes; D015415:Biomarkers / Q000032:analysis* / Q000378:metabolism; D004350:Drug Residues / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000662:veterinary*; D005399:Fishes / Q000378:metabolism*; D057166:High-Throughput Screening Assays / Q000379:methods / Q000662:veterinary*; D020128:Inhibitory Concentration 50; D015394:Molecular Structure; D009025:Morpholines / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D009581:Nitrofurans; D023303:Oxazolidinones / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D012680:Sensitivity and Specificity
C504349:3-amino-5-morpholinomethyl-2-oxazolidinone; D001547:Benzaldehydes; D015415:Biomarkers; D009025:Morpholines; D009581:Nitrofurans; D023303:Oxazolidinones; C043850:2-nitrobenzaldehyde; C007795:furaltadon