Quantitative Determination of β-Agonists in Pork Using the Amplified Luminescent Proximity Homogeneous Assay
  
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KeyWord:AlphaLISA  β-agonists  pork
  
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GONG Qian,XI Cun-xian,NIE Fu-ping,CAO Shu-rui,TANG Bo-bin,WANG Guo-min,CHEN Dong-dong,MU Zhao-de* 1.重庆医科大学药学院;2.重庆出入境检验检疫局重庆市进出口食品安全工程技术研究中心;3.重庆市生物化学与分子药理学重点实验室;4.中国检验检疫科学研究院
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Abstract:
      An amplified luminescent proximity homogeneous assay(AlphaLISA) was developed for the determination of four kinds of β agonists(clenbuterol,salbutamol,brombuterol,terbutaline) in pork.The analytes were stepwise extracted with ammonium acetate and β-glucuronidase/sulfatase,the extract was columned and concentrated to dryness,redisolved in buffer.Samples were reacted with botinylated antigen,antibody,donor beads and acceptor beads,and detected by AlphaLISA with matrix standard curve.As a result,there are good linear relationships in the concentration range of 0.1-500 ng/mL,with correlation coefficients larger than 0.98.The cross-reactivities for clenbuterol,brombuterol,terbutaline were 143.3%,179.2%and 95.6%,respectively,and the limits of detection(LOD) were 0.03-0.08 ng/mL.The average recoveries of four compounds at spiked levels of 0.5,10,20 μg/kg were in the range of 82.5%-115.9%with relative standard deviations(RSD) not less than 12.0%.The method is simple,rapid and accurate,and is suitable for the detection and screening of β-agonists in pork.
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