化学发光酶免疫法检测猪肉中氯丙嗪残留

采用棋盘滴定法确定包被抗原浓度和抗体稀释倍数,通过单因素实验优化了竞争反应时间、磷酸盐缓冲液浓度、甲醇含量、pH值等参数,建立了氯丙嗪的间接竞争化学发光酶免疫检测方法,并考察了方法的特异性、灵敏度和稳定性.结果表明,最佳反应条件为包被抗原浓度为0.05 μg/L;氯丙嗪抗体稀释32000倍；体系缓冲液为含10％甲醇、0.1 mol/L磷酸盐缓冲液(pH 7.0).本方法的IC50为0.12 μg/L;检出限为0.02 μg/L;线性范围0.02～24.78 μg/L;批内和批间相对标准偏差均小于10％.与其它结构类似物没有明显交叉反应.猪肉检测的平均回收率为87.4％～105.6％,与高效液相色谱方法的相关性良好.本方法具有较高的灵敏度和较好的稳定性.

Determination of Residues of Chlorpromazine in Pork by Chemiluminescent Enzyme Immunoassay

An indirect competitive chemiluminescent enzyme immunoassay(ic-CLEIA) method was developed for the determination of chlorpromazine.The optimal coating concentration and antibody dilution were determined by chequerboard titration.The effects of incubation time,ion concentration,methanol content and pH on the sensitivity of ic-CLEIA were investigated by single-factor experiments.The results showed that the optimized assay conditions were as follows: coating antigen concentration,0.05 μg/L;and dilution fold of antibody enzyme conjugate in 0.1 mol/L pH 7.0 phosphate buffered solution dilution(PBS) containing 10% methanol,32000.The developed method presented an IC50 of 0.12 μg/L,a detection limit of 0.02 μg/L,a linear range of 0.02 to 24.78 μg/L.Intra-and inter-batch relative standard deviations were less than 10%.There were no conspicuous cross reactions with other structural analogues of chlorpromazine.The analytical recovery in pork samples tissue was 87.4%-105.6%.A comparison result between high performance liquid chromatography(HPLC) and the developed assay showed better relativity.The method is sensitive and stable.