用于检测β2-肾上腺素能兴奋剂克伦特罗的酶免疫测定法。

Enzyme immunoassay for clenbuterol, an beta 2-adrenergic stimulant.

作者信息

Yamamoto I, Iwata K

出版信息

J Immunoassay. 1982;3(2):155-71. doi: 10.1080/15321818208056993.

DOI:10.1080/15321818208056993

Abstract

A sensitive double antibody and heterologous enzyme immunoassay for the quantitation of clenbuterol is established. Specific antiserum to this agent was raised in rabbits by immunization with diazotized clenbuterol and human serum albumin conjugate. For competitive reactions, antibody was incubated with a mixture of diazotized clenbuterol analog (NA 1141) labelled with beta-D-galactosidase and unlabelled standard or sample clenbuterol. The antibody-bound enzyme hapten was separated from free hapten by anti-rabbit IgG immobilized to a polystyrene ball. Activity of the enzyme on the solid phase was fluorometrically determined. The assay system made it possible to ascertain values as low as 0.5 pg /tube of clenbuterol. By use of this assay method, the time course of plasma levels of clenbuterol was examined after a single oral administration (20 micrograms) to 3 healthy volunteers. It was shown that the maximum level was achieved after 2-3 hr with approximately 10 ng clenbuterol/dl of plasma.

摘要

建立了一种用于定量检测克伦特罗的灵敏双抗体和异源酶免疫分析法。用重氮化克伦特罗与人血清白蛋白偶联物免疫家兔，制备了针对该药物的特异性抗血清。在竞争性反应中，将抗体与用β-D-半乳糖苷酶标记的重氮化克伦特罗类似物（NA 1141）和未标记的标准品或样品克伦特罗的混合物一起孵育。通过固定在聚苯乙烯球上的抗兔IgG将结合抗体的酶半抗原与游离半抗原分离。用荧光法测定固相上酶的活性。该检测系统能够测定低至0.5 pg/管的克伦特罗值。使用这种检测方法，对3名健康志愿者单次口服（20微克）克伦特罗后血浆中克伦特罗水平的时间进程进行了检测。结果表明，2-3小时后达到最高水平，血浆中克伦特罗约为10 ng/dl。