Comparison of two polyclonal antibodies for the detection of 19‐Nortestosterone in Bovine Bile by ELISA

Cooper, J.K.; Elliott, Christopher T.; Baxter, G. Andrew; Hewitt, S. Armstrong; McEvoy, J. D. G.; McCaughey, W.J.

doi:10.1080/09540109809354976

Cited by 11 publications

(3 citation statements)

References 11 publications

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“…The disparity in cross-reactivities is in agreement with a previous study carried out in this laboratory investigating the matrix effects of bovine bile on two polyclonal antibodies raised against 19-nortestosterone. 25 The strength of the interaction between an antibody and an antigen is known as the affinity. Polyclonal antisera contain complex mixtures of antibodies with different affinities, which are affected by temperature, pH and solvent.…”

Section: Discussionmentioning

confidence: 99%

Development of a rapid screening test for veterinary sedatives and the beta-blocker carazolol in porcine kidney by ELISA

Cooper¹,

Delahaut²,

Fodey³

et al. 2004

Analyst

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Sedatives and tranquillisers are frequently used to reduce stress during the transportation of food producing animals. The most widely used classes of sedatives include the butyrophenone azaperone, the phenothiazines acepromazine, propionylpromazine, chlorpromazine and the [small beta]-blocker, carazolol. For regulatory control purposes, tolerances for azaperone and carazolol have been set by the European Union as 100 and 25 [micro sign]g kg(-1), respectively. Furthermore, the use of the phenothiazines is prohibited and therefore has a zero tolerance. A method for the detection of residues of five tranquillisers and one [small beta]-blocker using a single ELISA plate has been developed. Kidney samples (2.5 g) were extracted with dichloromethane and applied to a competitive enzyme immunoassay using three polyclonal antibodies raised in rabbits against azaperol, propionylpromazine and carazolol conjugates. In sample matrix, the azaperol antibody cross-reacted 28.0% with azaperone and the propionylpromazine antibody cross-reacted 24.9% with acepromazine and 11.7% with chlorpromazine. In the ELISA, the detection capabilities of the six sedatives, azaperol, azaperone, carazolol, acepromazine, chlorpromazine, and propionylpromazine are 5, 15, 5, 5, 20 and 5 [micro sign]g kg(-1), respectively. The proposed method is a sensitive and rapid multi-residue technique that offers a cost effective alternative to current published procedures, without any concession on the ability to detect sedative misuse.

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Section: Discussionmentioning

confidence: 99%

Development of a rapid screening test for veterinary sedatives and the beta-blocker carazolol in porcine kidney by ELISA

Cooper¹,

Delahaut²,

Fodey³

et al. 2004

Analyst

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“…The standard curve was: y = -0.3194x + 1.6316, R 2 = 0.9927 The maximum signal of the assay was always around 1.5 units of absorbance. The sensitivity reached by this immunoassay proved suffi cient for analysing samples and superior to the previously reported immunoassay for NT (Cooper et al, 1998). The optimized immunoassay was evaluated to determine its selectivity by fi ve structurally related steroids (testosterone, medroxyprogesterone, epitestosterone, progesterone and dehydroepiandrosterone).…”

Section: Resultsmentioning

confidence: 99%

Development and optimization of an indirect enzyme-linked immunosorbent assay for 19-nortestosterone

Tian

Zhang

et al. 2008

J. Anim. Feed Sci.

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A polyclonal antibody-based enzyme-linked immunosorbent assay (ELISA) has been developed, optimized and validated to measure this anabolic steroid. Infl uence of several physicochemical parameters, such as incubation time, ionic strength, detergent concentration and pH were selected to provide a highest sensitivity on the ELISA format. The regression equation of the fi nal inhibition curve was: y = -0.3194x + 1.6316, R 2 = 0.9927. The linear range was between 0.1 and 25 ng/ml and the IC 50 was 3.5 ng/ml. The specifi city was evaluated by fi ve structurally related anabolic steroids, and none of them had signifi cant cross-reactivity. Finally, the accuracy and precision of this assay were evaluated by means of spiked samples. The recovery was between 76.9 and 104.7%, and the variation coeffi cient was between 5.2 and 13.4%.

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“…[4][5][6] To improve extraction, these conjugates should be deconjugated to compounds soluble in organic solvents prior to extraction. This is achieved by enzymatic hydrolysis 6,7 or chemical hydrolysis using methanolic hydrogen chloride, 8 techniques used prior to determination by immunoassay. Cooper et al 8 compared the two methods for diethylstilboestrol in bovine urine and found that, on the whole, enzymatic hydrolysis was more efficient than chemical hydrolysis.…”

Section: Introductionmentioning

confidence: 99%

Validation of a simple, sensitive method for the determination of β-estradiol in bovine urine using gas-chromatography negative-ion chemical ionization mass spectrometry

Akre

Fedeniuk

MacNeil

2004

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A new method is presented for the analysis of 17[small beta]-estradiol in bovine urine. After deconjugation, the sample is cleaned up using an OASIS[trade mark sign] HLB disposable cartridge and extracted into 1-chlorobutane. The hormone is derivatized using pentafluorobenzoyl chloride. The derivatized estradiol is quantitated using gas-chromatography negative-ion chemical ionization mass spectrometry. Calibrations, obtained using spiked blank urine, are linear in the range of 100-1000 pg mL(-1) with CC[small alpha] approximately 170 pg mL(-1) and CC[small beta] of 287 pg mL(-1). Recoveries are in the range of 80 to 130%. The method is rugged, rapid and sensitive when compared to other hormone methods.

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Comparison of two polyclonal antibodies for the detection of 19‐Nortestosterone in Bovine Bile by ELISA

Abstract: 19-Nortestosterone (ß-NT) is banned for use as a growth promoter in food animals

Cited by 11 publications

References 11 publications

Development of a rapid screening test for veterinary sedatives and the beta-blocker carazolol in porcine kidney by ELISA

Development of a rapid screening test for veterinary sedatives and the beta-blocker carazolol in porcine kidney by ELISA

Development and optimization of an indirect enzyme-linked immunosorbent assay for 19-nortestosterone

Validation of a simple, sensitive method for the determination of β-estradiol in bovine urine using gas-chromatography negative-ion chemical ionization mass spectrometry

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