Abstract
A high affinity polyclonal antibody-based enzyme linked immunosorbent assay (ELISA) was developed for the quantification of zeranol in bovine urine. On the basis of urine matrix studies, the optimized dilution factors producing insignificant matrix interference were selected as 1:5 in pretreatment. In the improved ELISA, the linear response range was between 0.02 and 1 μg/ml, and the detection limit was 0.02 μg/ml for the assay. The overall recoveries and the coefficients of variation (CVs) were in the range of 82%∼127% and 3.5%∼8.8%, respectively. Thirty-six bovine urine samples spiked with zeranol (ranging from 0.2 to 10 μg/ml) were detected by the ELISA and liquid chromatography (LC) method, and good correlations were obtained between the two methods (R 2=0.9643). We conclude that this improved ELISA is suitable tool for a mass zeranol screening and can be an alternative for the conventional LC method for zeranol in bovine urine.
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Baldwin, R.S., Williams, R.D., Terry, M.K., 1983. Zeranol: a review of the metabolism, toxicology, and analytical methods for detection of tissue residues. Regul. Toxicol. Pharmacol., 3(1):9–25. [doi:10.1016/0273-2300(83)90046-6]
Chen, X.J., Liu, H.C., Meng, F.J., 1990. Direct enzyme-linked immunoassay for zearalenone. Plant Physiol. Commun., 16(1):70–76 (in Chinese).
Daniel, T., Robert, F.M., 1983. Radioimmunoasssay for zearalenone and zearalanol in human serum: production, properties and use of porcine antibodies. Appl. Environ. Microbiol., 45(1):16–23.
Dickson, L.C., MacNeil, J.D., Reid, J., Fesser, A.C., 2003. Validation of screening method for residues of diethylstilbestrol, dienestrol, hexestrol, and zeranol in bovine urine using immunoaffinity chromatography and gas chromatography/mass spectrometry. J. AOAC Int., 86(4):631–639.
Dioxn, S.N., Russell, K.L., 1983. Radioimmunoassay of the anabolic agent zeranol II. Zeranol concentrations in urine of sheep and cattle implanted with zeranol (Ralgro). J. Vet. Pharmacol. Therap., 6:173–179.
Dong, J., Liu, X.J., Han, Z.J., 2001. Hapten design and antibodies preparation of fipronil. Jiangsu J. Agric. Sci., 17(3):172–175 (in Chinese).
Fang, X., Chen, J., Guo, D., Wang, G., 2002. Detection and identification of zeranol in chicken or rabbit liver by liquid chromatography-electrospray tandem mass spectrometry. J. AOAC Int., 85(4):841–847.
Fang, X.M., Chen, J.H., Tang, Y.F., 2003. Determination of zeranol in chicken livers by high performance liquid chromatography with ultraviolet detection. Chim. J. Chromatogr., 21(2):158–161 (in Chinese).
Horie, M., Nakazawa, H., 2000. Determination of trenbolone and zeranol in bovine muscle and liver by liquid chromatography-electrospray mass spectrometry. J. Chromatogr. A, 882(1–2):53–62. [doi:10.1016/S0021-9673(00)00205-3]
Jansen, E.H.J.M., Vanden Berg, R.H., Zomer, G., Willemsen, C.E., Stephany, R.W., 1986. A chemiluminescent immunoassay for zeranol and its metabolites. J. Vet. Pharmacol. Therap., 9:101–108.
Liu, Y., Liu, X.J., Yu, X.Y., Zhang, C.Z., Gong, Z.M., Zhou, Q., 2006. Synthesis of artificial antigen and preparation of polyclonal antibodies against zeranol. J. Anal. Sci., 22(1):1–4 (in Chinese). [doi:10.2116/analsci.22.1]
Richardson, K.E., Hagler, W.M., Mirocha, C.J., 1985. Production of zeralenone, α-and β-zearalenol and α-and β-zearalanol by Fusarium spp. in rice culture. J. Agric. Food Chem., 33(5):862–866. [doi:10.1021/jf00065a024]
Sorensen, L.K., Elbaek, T.H., 2005. Determination of mycotoxins in bovine milk by liquid chromatography tandem mass spectrometry. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci., 820(2):183–196. [doi:10.1016/j.jchromb.2005.03.020]
Taguchi, S., Yoshida, S., Tanaka, Y., Hori, S., 2001. Simple and rapid analysis of trenbolone and zeranol residues in cattle muscle and liver by stack-cartridge solid-phase extraction and HPLC using on-line clean-up with EC and UV detection. Shokuhin Eiseigaku Zasshi, 42(4):226–230 (in Japanese).
Tuomola, M., Cooper, K.M., Lahdenperä, S., Andrew Baxter, G., Elliott, C.T., Glenn Kennedy, D., Lövgren, T., 2002. A specificity-enhanced time-resolved fluoroimmunoassay for zeranol employing the dry reagent all-in-one-well principle. Analyst, 127:83–86. [doi:10.1039/b108671p]
Wang, H.J., Chao, Z., Sheng, J.Z., 2004. Preparation and identification of monoclonal antibody to zeranol. Chin. J. Vet. Med., 40(10):3–5 (in Chinese).
Wortberg, M., Kreissig, S.B., Jones, G., Rocke, D.M., Hammock, B.D., 1995. An immunoarray for simultaneous determination of multiple triazine herbicides. Analytica Chimica Acta, 304(3):339–352.
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Project supported by the National Natural Science Foundation of China (No. 30471155) and the Agriculture Key Technologies R & D Program of Shanghai (No. (2003) 9-4), China
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Liu, Y., Zhang, Cz., Yu, Xy. et al. Development and evaluation of immunoassay for zeranol in bovine urine. J. Zhejiang Univ. - Sci. B 8, 900–905 (2007). https://doi.org/10.1631/jzus.2007.B0900
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DOI: https://doi.org/10.1631/jzus.2007.B0900