Stanley S, Jeganathan A, Wood T, Henry P, Turner S, Woods W E, Green M, Tai H H, Watt D, Blake J
Maxwell H. Gluck Equine Research Center, University of Kentucky, Lexington 40546-0099.
J Anal Toxicol. 1991 Nov-Dec;15(6):305-10. doi: 10.1093/jat/15.6.305.
We have raised antibodies to morphine and etorphine and developed one-step enzyme-linked immunosorbent assays (ELISA) for these drugs as part of a panel of post race tests for drugs in racing horses. These tests are simple, can be completed in 2 h, and can be read by visual inspection. The morphine ELISA has an I50 for morphine of about 1.5 ng/mL, while the etorphine ELISA has an I50 for etorphine of 250 pg/mL. Cross-reactivity studies show that the antimorphine antibody cross-reacts well with levorphanol, hydromorphone, and oxycodone, while the anti-etorphine antibody showed no cross-reactivity with buprenorphine, diprenorphine, oxymorphone, morphine, or thebaine. The morphine test readily detected parent morphine or its metabolites in equine urine for at least 8 h after administration of 50 mg/horse, while a 0.1 micrograms/kg dose of etorphine was detectable for up to 48 h post dosing. For each test the background activity in post-race urines was equal to or less than the I50 for the standard curves, making them useful equine forensic tests. Each of these tests has detected "positives" in post race urine samples and as such these tests are capable of substantially improving the speed and efficacy of both pre-race and post-race testing for morphine, etorphine, and their congeners in racing horses.
我们制备了针对吗啡和埃托啡的抗体,并开发了针对这些药物的一步法酶联免疫吸附测定(ELISA),作为赛马赛后药物检测项目的一部分。这些检测方法简单,可在2小时内完成,且通过目视检查即可读取结果。吗啡ELISA对吗啡的半数抑制浓度(I50)约为1.5 ng/mL,而埃托啡ELISA对埃托啡的I50为250 pg/mL。交叉反应研究表明,抗吗啡抗体与左啡诺、氢吗啡酮和羟考酮有良好的交叉反应,而抗埃托啡抗体与丁丙诺啡、二丙诺啡、羟吗啡酮、吗啡或蒂巴因无交叉反应。吗啡检测能够在每匹马注射50 mg后至少8小时内轻松检测到马尿中的母体吗啡或其代谢物,而0.1微克/千克剂量的埃托啡在给药后长达48小时内均可检测到。对于每项检测,赛后尿液中的背景活性等于或低于标准曲线的I50,这使得它们成为有用的马法医检测方法。这些检测方法均在赛后尿液样本中检测到了“阳性”结果,因此,这些检测方法能够大幅提高赛马赛前和赛后对吗啡、埃托啡及其同类物检测的速度和效率。