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 facto...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 altemative for the conventional LC method for zeranol in bovine urine.展开更多
Zeranol (Z) is an estrogenic growth-promoting agent synthesized from mycotoxin zearalenone (Zen). Inadvertent consumption of Z and its structural analogs from meat or grain products remain a food safety concern. An ec...Zeranol (Z) is an estrogenic growth-promoting agent synthesized from mycotoxin zearalenone (Zen). Inadvertent consumption of Z and its structural analogs from meat or grain products remain a food safety concern. An economic and rapid high performance liquid chromatography method with electrochemical detection using disposable screen-printed carbon electrode is developed for determination of Z, Zen and 3 major metabolic analogs α-zearalenol (α-Ze), β-zearalenol (β-Ze), and β-zearalanol (β-Za). The electrochemical method was validated for application in food matrices including beef, pork, feed and cereal after optimized liquid and/or solid-phase extraction procedures. All 5 Z analogs were separated in 10 minutes with the limits of detection ranging from 15 ng/ml for α-Ze and 25 ng/ml for Z and Zen;the limit of quantitation ranged from 40 50 ng/ml. The recoveries were all above 75% regardless of matrix types and extraction procedures. The intra and inter day variations were both less than 6% at the nominal concentration of 1 μg/ml and less than 13% at 100 ng/ml level. Chromatographically time-matched peaks of Z, α-Ze and β-Za were observed in moldy feed, cereal and rice with high productivity, indicating possible grain-specific Zs exposure for animals and human. Proper exercise of preservative procedures for grain and grain products to prevent it from mold production is imperative. The simplicity and reproducibility of this method affords quick and reliable quantitation of multiple types of Z analogs in food products and can offer semi-confirmative information comparable to UV detection and supplementary to ELISA screening.展开更多
基金Project supported by the National Natural Science Foundation of China (No. 30471155) the Agriculture Key Technologies R & D Program of Shanghai (No. (2003) 9-4), China
文摘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 altemative for the conventional LC method for zeranol in bovine urine.
文摘Zeranol (Z) is an estrogenic growth-promoting agent synthesized from mycotoxin zearalenone (Zen). Inadvertent consumption of Z and its structural analogs from meat or grain products remain a food safety concern. An economic and rapid high performance liquid chromatography method with electrochemical detection using disposable screen-printed carbon electrode is developed for determination of Z, Zen and 3 major metabolic analogs α-zearalenol (α-Ze), β-zearalenol (β-Ze), and β-zearalanol (β-Za). The electrochemical method was validated for application in food matrices including beef, pork, feed and cereal after optimized liquid and/or solid-phase extraction procedures. All 5 Z analogs were separated in 10 minutes with the limits of detection ranging from 15 ng/ml for α-Ze and 25 ng/ml for Z and Zen;the limit of quantitation ranged from 40 50 ng/ml. The recoveries were all above 75% regardless of matrix types and extraction procedures. The intra and inter day variations were both less than 6% at the nominal concentration of 1 μg/ml and less than 13% at 100 ng/ml level. Chromatographically time-matched peaks of Z, α-Ze and β-Za were observed in moldy feed, cereal and rice with high productivity, indicating possible grain-specific Zs exposure for animals and human. Proper exercise of preservative procedures for grain and grain products to prevent it from mold production is imperative. The simplicity and reproducibility of this method affords quick and reliable quantitation of multiple types of Z analogs in food products and can offer semi-confirmative information comparable to UV detection and supplementary to ELISA screening.
