Study On Chemilumineseent Immunosorbent Assay For17β-19-Nortestosterone

In this paper, a chemical luminescence immunoassay for nandrolone was established and it can simultaneously detect four other anabolic hormones, such as trenbolone, propionic nandrolone, stanolone, mestanolone.On the basis of nandrolone polyclonal antibody,main parameters of chemiluminescence immunoassay were optimized, including the antibody-coating amount, concentration of enzyme labeled antigen, type and concentration of the sample dilution and luminescent substrate. The performance of the established chemiluminescent immunoassay for nandrolone was systematically evaluated. This method can simultaneously detect nandrolone, trenbolone, nandrolone propionate, stanolone, mestanolone five kinds of anabolic hormones, with high sensitivity, which were nandrolone IC50=0.050μg/L, IC15=0.00378μg/L, trenbolone IC50=0.094μg/L, IC15=0.00330μg/L, propionate, nandrolone IC50=0.66μg/L, IC15=0.259μg/L, Stanolone IC50=1.40μg/L, ICis=0.0398μg/L, Mestanolone IC50=2.40μg/L, IC15=0.078μg/L, compared with conventional ELISA analysis, sensitivity is increased by5to8times, the coefficient of variation of the detection method were less than20%.Pork, chicken and beef were selected as test samples, extracted by methanol, diluted by phosphate buffer solution, matrix effect was then eliminated. The average recovery of nandrolone from actual samples were80to100%, indicating that the accuracy of the method. Compared with the test results by the traditional ELISA, the results were correlated well, with the correlation coefficient R2=0.9988; further use of high performance liquid chromatography for the validation of the accuracy of nandrolone chemiluminescent immunoassay, the results of two methods has high linear correlation coefficient (R2=0.9816), indicating that the established nandrolone chemiluminescent immunoassay can be used for the rapid detection of nandrolone residue in animal-derived foods.