竞争化学发光酶免疫测定技术检测盐酸克伦特罗

Determination of clenbuterol by competitive chemiluminescent enzyme immunoassay

中文关键词:  竞争化学发光酶免疫测定法  竞争酶联免疫吸附测定法  盐酸克伦特罗

英文关键词:competitive chemiluminescent enzyme immunoassay  competitive enzyme linked immunosorbent assay  clenbuterol

中文摘要:

      目的 建立一种检测盐酸克伦特罗(clenbuterol, CLB)的竞争化学发光酶免疫测定法(chemiluminescent enzyme immunoassay, CLEIA)。方法 基于CLB的竞争酶联免疫吸附测定法(enzyme linked immunosorbent assay, ELISA), 通过优化反应温度、反应时间和缓冲体系pH建立了CLB的CLEIA测定法。用CLB小鼠中毒模型评价CLEIA检测法对实际样本的检测效果, 并将CLEIA与LC-MS法的相关性进行了比较。结果 CLEIA法测定CLB的检测范围为0.0433~42.5363 μg/L, 检测限为0.0178 μg/L, 变异系数小于10%, 回收率为95%~110%, 对莱克多巴胺和沙丁胺醇仅有极低的交叉反应性。所建立的CLEIA法与LC-MS法在检测实际样本时具有较高的相关性。结论 本研究建立的CLEIA法操作简便, 检测限、灵敏度、准确度、特异性均符合检测要求, 可用于实际样品的检测。

英文摘要:

      Objective To establish a method for the determination of clenbuterol (CLB) by competitive chemiluminescent enzyme immunoassay assay (CLEIA). Methods The CLEIA method was established for the detection of CLB after the optimization of reaction temperature, reaction time and buffer system pH based on enzyme linked immunosorbent assay (ELISA). The effect of CLEIA test on the actual samples was evaluated by CLB poisoning model of mice. And the correlation between CLEIA and LC-MS was compared. Results The detection range of CLB by CLEIA was 0.0433~42.5363 μg/L with the detection limit was 0.0178 μg/L, variation coefficient less than 10%. The recoveries of method were ranged from 95% to 110%. There was little cross-reaction to ractopamine and albuterol in CLB detection by CLEIA. Moreover, the established CLEIA method had a high correlation with LC-MS method in the detection of actual samples. Conclusion This CLIEA method is simple, sensitive, accurate and it has a lower detection limit and good specificity, which is suitable for the rapid determination of CLB in samples.