Development and evaluation of immunoassay for zeranol in bovine urine

Liu, Yuan; Zhang, Cunzhen; Yu, Xiangyang; Zhang, Zhiyong; Zhang, Xiao; Liu, Rong-rong; Liu, Xianjin; Gong, Zhaoning

doi:10.1631/jzus.2007.b0900

Cited by 46 publications

(33 citation statements)

References 13 publications

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“…Competitive inhibition curves against NOR were established and the general assay procedures were described previously (Liu et al, 2007;Wu et al, ). Based on the optimized concentrations, an indirect competitive ELISA (icELISA) method was developed, and the calibration curve was fitted based on the average of three separate assays in triplicate.…”

Section: Development Of Indirect Competitive Elisa Standard Curvesmentioning

confidence: 99%

Development of a class-specific polyclonal antibody-based indirect competitive ELISA for detecting fluoroquinolone residues in milk

Fan¹,

Yang²,

Jiang³

et al. 2012

J. Zhejiang Univ. Sci. B

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Modified 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) method was employed to synthesize the artificial antigen of norfloxacin (NOR), and New Zealand rabbits were used to produce anti-NOR polyclonal antibody (pAb). Based on the checkerboard titration, an indirect competitive enzyme-linked immunosorbent assay (icELISA) standard curve was established. This assay was sensitive and had a working range from 0.12 to 68.40 ng/ml, with the half maximal inhibitory concentration (IC 50 ) and limit of detection (LOD) values of 2.7 ng/ml and 0.06 ng/ml, respectively. The produced pAb exhibited high cross-reactivity to fluoroquinolones (FQs) tested, and the IC 50 values to enoxacin, ciprofloxacin, and pefloxacin were 3.1, 3.4, and 4.1 ng/ml, respectively. It also indicated that the concentrations of NaOH and methanol in assay buffer should not be higher than 10% and 30%. When spiked in milk at 5, 20, and 50 ng/ml, the recoveries for NOR, enoxacin, ciprofloxacin, and pefloxacin ranged 90.5%-98.0%, 84.0%-95.2%, 94.0%-106.0%, and 89.5%-100.0%, respectively. The results suggest that this class-specific pAb-based icELISA could be utilized for the primary screening of FQ residues in animal-original products.

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Section: Development Of Indirect Competitive Elisa Standard Curvesmentioning

confidence: 99%

Development of a class-specific polyclonal antibody-based indirect competitive ELISA for detecting fluoroquinolone residues in milk

Fan¹,

Yang²,

Jiang³

et al. 2012

J. Zhejiang Univ. Sci. B

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“…The presence of ZAL and its metabolites in cattle urine is often used as a marker for the deliberate use of ZAL . Currently, several methods have been developed for the detection of the residues of ZAL and/or their metabolites in a large variety of matrices, such as thin‐layer chromatography , gas chromatography–mass spectrometry , high‐performance liquid chromatography (HPLC) , liquid chromatography tandem mass spectrometry (LC‐MS/MS) and immunoassay . The matrices reported in previous research included different animal (chicken, bovine and sheep) tissues (liver, kidney and muscle) , urine , milk , hair and water .…”

Section: Introductionmentioning

confidence: 99%

“…In contrast, immunoassay is comparably efficient and can be applied in large‐scale screening. Accordingly, enzyme‐linked immunosorbent assay (ELISA) and chemiluminescent enzyme immunoassay (CLEIA) were developed for the determination of ZAL . CLEIA has become very popular in recent years in food safety and environmental analysis due to its high sensitivity and wide dynamic range .…”

Section: Introductionmentioning

confidence: 99%

Determination of zeranol and its metabolites in bovine muscle and liver by a chemiluminescence enzyme immunoassay: compared to an ultraperformance liquid chromatography tandem mass spectroscopy method

Jiang

Wang²,

Zhu

et al. 2013

Luminescence

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A chemiluminescent enzyme immunoassay (CLEIA) was compared to an ultraperformance liquid chromatography tandem mass spectroscopy (UPLC-MS/MS) procedure for the analysis of zeranol and its metabolites in bovine tissue samples. Apparent recoveries from fortified samples by both methods were comparable at 0.5-4.0 µg/kg and a significant correlation was obtained. For CLEIA analysis, hapten mimicking the analyte was first synthesized and conjugated with the carrier protein bovine serum albumin as the immunogen to produce monoclonal antibody. The obtained antibody showed extensive cross-reactivity toward zeranol metabolites (zearalanone). The limit of detection of CLEIA and UPLC-MS/MS was 0.05 µg/kg and 0.5 µg/kg, respectively. Recoveries of both methods for fortified samples were higher than 75.0% with the coefficient of variation less than 15%. These results indicated that the combination of screening with CLEIA and confirmation with UPLC-MS/MS for zeranol and its metabolites would be a reliable method for a large number of bovine samples.

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“…Liu et al 2007). CLEIA utilizes an enzyme-labeled antigen or antibody and enzyme-catalyzing chemiluminescent reagents to emit light.…”

Section: Introductionmentioning

confidence: 99%

Micro-Plate Chemiluminescence Enzyme Immunoassay for Determination of Zeranol in Bovine Milk and Urine

et al. 2012

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Zeranol (a-Zearalanol, a-ZAL), well-known as an anabolic promoter, was officially banned in Europe due to its potential carcinogenic and endocrine-disrupting biological activities. In this study, a method for the determination of zeranol residues in bovine milk and urine was developed based on micro-plate chemiluminescence enzyme immunoassay (CLEIA). The limit of detection (LOD) was 50 ng/L, and the linear range was between 100 ng/L and 4510 ng/L, with a correlation coefficient (R 2 )0.9982. The recoveries of zeranol in bovine milk and urine were between 84.7% and 123.6%. This study showed that CLEIA was a reliable, convenient, and sensitive method for screening zeranol residues in bovine milk and urine.

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Development and evaluation of immunoassay for zeranol in bovine urine

Cited by 46 publications

References 13 publications

Development of a class-specific polyclonal antibody-based indirect competitive ELISA for detecting fluoroquinolone residues in milk

Development of a class-specific polyclonal antibody-based indirect competitive ELISA for detecting fluoroquinolone residues in milk

Determination of zeranol and its metabolites in bovine muscle and liver by a chemiluminescence enzyme immunoassay: compared to an ultraperformance liquid chromatography tandem mass spectroscopy method

Micro-Plate Chemiluminescence Enzyme Immunoassay for Determination of Zeranol in Bovine Milk and Urine

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