Midha K K, Hubbard J W, Cooper J K, Mackonka C
J Pharm Sci. 1983 Jul;72(7):736-9. doi: 10.1002/jps.2600720705.
Haptens were prepared by the reaction of d-ephedrine or l-ephedrine with methyl acrylate and subsequent alkaline hydrolysis of the methyl ester groups. The haptens were coupled to bovine serum albumin by a mixed anhydride method, and the resulting drug-protein conjugates were used to immunize rabbits. Antisera raised to these conjugates were highly stereospecific. Neither antiserum cross-reacted with the optical antipode of its substrate nor with racemic pseudoephedrine. Separate radioimmunoassays (RIAs), developed for d-ephedrine and l-ephedrine, were used to measure the concentrations of the enantiomers of ephedrine in the blood of two volunteers dosed with racemic ephedrine. The RIAs were validated by comparing the sum of the concentrations of the enantiomers, determined by RIA, with total ephedrine concentrations determined by a nonstereoselective GLC-ECD method.
通过将d-麻黄碱或l-麻黄碱与丙烯酸甲酯反应，随后对甲酯基团进行碱性水解来制备半抗原。半抗原通过混合酸酐法与牛血清白蛋白偶联，所得的药物-蛋白质缀合物用于免疫兔子。针对这些缀合物产生的抗血清具有高度的立体特异性。两种抗血清均不与其底物的旋光对映体发生交叉反应，也不与外消旋伪麻黄碱发生交叉反应。分别针对d-麻黄碱和l-麻黄碱开发的放射免疫分析法（RIA），用于测量两名服用外消旋麻黄碱的志愿者血液中麻黄碱对映体的浓度。通过将RIA测定的对映体浓度之和与非立体选择性气相色谱-电子捕获检测法（GLC-ECD）测定的总麻黄碱浓度进行比较，对RIA进行了验证。
