猪肝和猪尿中沙丁胺醇和克伦特罗残留的酶联免疫吸附检测法研究

Detection of Salbutamol and Clenbuterol in Swine Liver and Swine Urine by Enzyme Linked Immunosorbent Assays

采用混合酸酐法将沙丁胺醇与牛血清白蛋白偶联,并免疫家兔制备抗沙丁胺醇抗血清。以间接ELISA法测定血清效价,测得抗血清最佳工作浓度为1∶6400,该抗沙丁胺醇抗血清对克伦特罗有110%的交叉反应性。建立了能够同时检测SAL和CL的间接竞争ELISA方法,该方法对沙丁胺醇的检测范围为0148ng/mL～8.0μg/mL,对克伦特罗的检测范围为0143ng/mL～7.3μg/mL,对沙丁胺醇的检测灵敏度为0148ng/mL,对克伦特罗的检测灵敏度为0143ng/mL。在猪肝和猪尿中添加1～50ng/g(ng/mL)的沙丁胺醇,添加回收率为72.2%～8418%,变异系数为4.2%～28.3%,在猪肝脏和猪尿中添加1～50ng/g(ng/mL)的克伦特罗,添加回收率为8413%～103.4%,变异系数为013%～35.5%。

The β 2-agonist, salbutamol (SAL) was linked to bovine serum albumin(BSA) using the mixed anhydride method and the anti-SAL serum was acquired in rabbits against the conjugate. The optional dilution of the anti-SAL serum was 1∶6 400 by the indirect enzyme linked immunosorbent assay (ELISA) and the anti-SAL serum has the cross-reactivity of 110% with Clenbuterol (CL). Then an indirect competitive ELISA, which can detect SAL and CL simultaneously, was developed. The limit of detection for SAL was 0.48 ng/mL with the range of detection 0.48 ng/mL^8.0 μg/mL and the limit of detection for CL was 0.43 ng/mL with the range of detection 0.43 ng/mL^7.3 μg/mL. The recovery rates ranged from 72.2% to 84.8% for detecting SAL fortified level of 1~50 ng/g (ng/mL) in swine liver and swine urine, the coefficients of variation was 4.2%~28.3%; the recovery rates ranged from 84.3% to 103.4% for detecting CL fortified level of 1~50 ng/g (ng/mL) in swine liver and swine urine, the coefficients of variation was 0.3%~35.5%.