Determination of 19-nortestosterone residues in aquaculture tissues by enzyme-linked immunosorbent assay and comparison with liquid chromatography and tandem mass spectrometry |
| |
Authors: | Chuan-Lai Xu Chi-fang Pen Kai Hao Zheng-yu Jin Xiao-gang Chu |
| |
Affiliation: | 1. School of Food Science and Technology , Southern Yangtze University , 214036, WuXi, JiangSu Province, China xcl@sytu.edu.cn;3. School of Food Science and Technology , Southern Yangtze University , 214036, WuXi, JiangSu Province, China |
| |
Abstract: | 19-Nortestosterone (17β-NT) was oximated by carboxymethoxylamine and then coupled with bovine serum albumin (BSA) in a mixed-anhydride reaction in order to produce an antibody. The conjugate rate of 17β-NT and BSA was estimated to be 24 by ultraviolet spectrophotometry. Polyclonal antibody of 17β-NT was acquired from the animal immunized with the conjugate. Through an indirect enzyme-linked immunosorbent assay (ELISA), which demonstrated that the synthesis of immunogen was successful, the titre of antiserum was found to be 6.4?×?105. Based on the purified antibody, a competitive indirect ELISA was developed. ELISA revealed that the limit of detection (LOD) was 0.07?ng?g?1, the recovery (in edible tissues) was 71–89%, and the working range was 0.05–31.25?ng?g?1. The preliminary evaluation of assay performance through specificity, sensitivity, precision, and accuracy revealed that this ELISA method could be used in the practical detection of 17β-NT in tissue samples. Moreover, this method was compared with high-performance liquid chromatography tandem mass spectrometry, for which the transition for quantification of 17β-NT was 275.4/109.1. |
| |
Keywords: | Determination 19-Nortestosterone Residues Aquaculture Enzyme-linked immunosorbent assay (ELISA) Liquid chromatography and tandem mass spectrometry |
|
|