An enzyme immunoassay for the anabolic agent zeranol

Patel, Poulam M.; Sn, Dixon

doi:10.1080/02652038909373742

Cited by 9 publications

(4 citation statements)

References 17 publications

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“…Fractions of 0.75 min were collected, the ZEA-oxime was detected photometrically at 274 nm. Fractions containing the derivative (30)(31)(32)(33)(34)(35)(36)(37) were pooled, and the solvent evaporated. An activated ester (18) waq formed with ZEA-oxime (5.25mg) by reaction with NHS (10mg) and DCC (35mg) in DMF (0.3ml) for 16 h at room temperature.…”

Section: Preparatton Of the Zearalenone Immunoreagentsmentioning

confidence: 99%

“…In order t o provide rapid and simple methods t o assay mycotoxins in foods and feeds that do n o t require extensive laboratory equipment, immunoassays have been shown t o be good alternatives t o conventional physico-chemical methods (23). Enzyme immunoassays (EIA) have been described for D O N -t y p e (4, 17, 26, 40,43) and ZEA-type (9, 21, 30,41) mycotoxins. I n this report w e describe the development of a highly sensitive EIA specific for ZEA-type mycotoxins.…”

Section: Introductionmentioning

confidence: 99%

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Studies on the Application of Enzyme Immunoassays for the Fusarium Mycotoxins Deoxynivalenol, 3‐Acetyldeoxynivalenol, and Zearalenone

Usleber

Renz

Märtlbauer

et al. 1992

Journal of Veterinary Medicine, Series B

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Polyclonal antisera against zearalenone (ZEA) were produced in rabbits after immunization with ZEA-oxime coupled to human serum albumin. Using these antibodies and a ZEA-oxime-horseradish peroxidase conjugate in a competitive direct enzyme immunoassay (EIA), the detection limit for ZEA was 70 pg/ml. The relative cross-reactivities of the assay with ZEA, a-zearalenol, p-zearalenol, zearalanone, a-zearalanol, and p-zearalanol, respectively, were 100 Yo, 37.3 Yo, 7.2 %, 59.2 Yo, 5.3 %, and 3.9%, respectively. This EIA and two EIAs for deoxynivalenol (DON) and 3-acetyldeoxynivalenol(3-AcDON) (USLEHER et al., 1991) were used to analyze wheat samples. The limits of determination for DON, 3-AcDON, and ZEA in wheat were 200ppb, 50ppb, and 20ppb, respectively. The analysis of reference materials (wheat flour) containing D O N by EIA showed good agreement with the nominal values. The EIA for ZEA was in addition used to analyze biological fluids, obtained during a feeding trial. Two lactating cows were administered 25 mg and 100 mg ZEA per day, respectively, over a period of 6 days. Serum, milk, urine, and feces were assayed in the ZEA-EIA with and without sample treatment with P-glucuronidase prior to the analysis. Maximum toxin levels (ZEA-equivalents) found in milk were 0.4 and 1.2 ppb (glucuronides). The toxin concentration in milk decreased rapidly after the last toxin administration. In the urine, maximum levels of toxinglucuronide conjugates were 23 ppb and 24 ppb, respectively. The serum toxin levels corresponded to those found in milk. In the feces, mean values were 150ppb and 500ppb, respectively, no conjugated toxins were found in feces.

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Section: Preparatton Of the Zearalenone Immunoreagentsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Studies on the Application of Enzyme Immunoassays for the Fusarium Mycotoxins Deoxynivalenol, 3‐Acetyldeoxynivalenol, and Zearalenone

Usleber

Renz

Märtlbauer

et al. 1992

Journal of Veterinary Medicine, Series B

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“…The absence of reactivity against a-zearalenol is of particular importance because antibodies used in past methods have always shown significant cross-reactivity against this compound, typically ranging from 21 to 54%. 6,7,15,19,20 With the exception of taleranol, the reactivity profile was slightly better than that obtained in an ELISA system also employing the specificity-enhanced zeranol antibody. 14 The cross-reactivity against taleranol was three times higher in the current study but still in the range found in some other zeranol antibodies.…”

Section: Resultsmentioning

confidence: 85%

“…3 A number of immunochemical methods have been developed for the screening of zeranol. [4][5][6][7] However, all the current immunochemical tests exhibit cross-reactivity to a greater or lesser extent with zearalenone, a-zearalenol and b-zearalenol, the three principal toxins produced by Fusarium spp. fungi.…”

Section: Introductionmentioning

confidence: 99%

A specificity-enhanced time-resolved fluoroimmunoassay for zeranol employing the dry reagent all-in-one-well principle

Tuomola

Cooper

Lahdenperä

et al. 2001

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A simple dry chemistry time-resolved fluorescence immunoassay (TR-FIA) method was developed for the measurement of zeranol in bovine urine samples. The samples were purified by immunoaffinity chromatography and a specificity-enhanced zeranol antibody was employed in the immunoassay. This resulted in a highly selective method, which had only negligible reactivity with Fusarium spp. toxins. The all-in-one-well dry chemistry concept made the assay very simple to use because all the assay-specific reagents were already present in the reaction wells in dry form. Only the addition of diluted sample extract was required to perform the competitive one-step TR-FIA and the results were available in less than 1 h. The analytical limit of detection (mean + 3s) for the immunoassay was 0.16 ng ml(-1) (n = 12) and the functional limit of detection for the whole method, estimated by the analysis of zeranol-free samples, was 1.3 ng ml(-1) (n = 20). The recovery of zeranol at the level of 2 ng ml(-1) was 99% (n = 18) and the within-assay variation ranged between 4.5 and 9.0%.

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