Kohno T, Murasugi N, Sakurai H, Watabe K, Nakamuta H, Koida M, Sugie Y, Ogouchi T, Inoue T, Yanaka M, Nomura M, Yanagawa A
Department of Environmental Health Sciences, Faculty of Pharmaceutical Sciences, Setsunan University, Osaka, Japan.
J Clin Lab Anal. 1998;12(6):356-62. doi: 10.1002/(sici)1098-2825(1998)12:6<356::aid-jcla5>3.0.co;2-#.
A sandwich transfer enzyme immunoassay for elcatonin (ECT) and its usability for the pharmacokinetic study are described. The anti-salmon calcitonin (SCT) antibody was used for the present assay. The assay procedure consisted of the reaction of ECT with 2,4-dinitrophenylbiotinyl anti-SCT IgG and anti-SCT Fab'-beta-D-galactosidase conjugate, trapping onto (anti-2,4-dinitrophenyl bovine serum albumin) IgG-coated polystyrene balls, eluting with epsilonN-2,4-dinitrophenyl-L-lysine and transferring to streptavidin-coated polystyrene balls and fluorometric detection of beta-D-galactosidase activity. The practical detection limit of ECT was 0.15 pg (44 amol)/50 microl of sample and 3 pg/ml as the concentration. The application of this method has enabled us to directly estimate the bioavailability of ECT dosed intranasaly at a therapeutic level (100 IU, 17 microg) for its anti-osteoporotic effect as compared to an intramuscular dose (40 IU, 6.7 microg). The pharmacokinetic parameters of the intranasal ECT (n = 6) thus estimated were as follows: the area underthe serum concentration-time curve (AUC) = 2,570 +/- 1,650 (SD) pg x min/ml, and the maximal concentration (Cmax) = 60 +/- 25 (SD) pg/ml with the maximal time (Tmax) = 17.5 +/- 6.9 (SD) min, when the AUC for the intramuscular ECT (n = 9) = 9,460 +/- 5,870 (SD) pg x min/ml and the Cmax = 165 +/- 79 (SD) pg/ml with the Tmax = 16.1 +/- 4.2 (SD) min.
描述了一种用于降钙素（ECT）的夹心转移酶免疫测定法及其在药代动力学研究中的实用性。本测定使用抗鲑鱼降钙素（SCT）抗体。测定程序包括ECT与2,4-二硝基苯基生物素化抗SCT IgG和抗SCT Fab'-β-D-半乳糖苷酶结合物的反应，捕获到（抗2,4-二硝基苯基牛血清白蛋白）IgG包被的聚苯乙烯球上，用εN-2,4-二硝基苯基-L-赖氨酸洗脱并转移到链霉亲和素包被的聚苯乙烯球上，以及对β-D-半乳糖苷酶活性进行荧光检测。ECT的实际检测限为0.15 pg（44 amol）/50微升样品，浓度为3 pg/ml。与肌肉注射剂量（40 IU，6.7微克）相比，该方法的应用使我们能够直接评估治疗水平（100 IU，17微克）鼻内给药的ECT的抗骨质疏松作用的生物利用度。由此估计的鼻内ECT（n = 6）的药代动力学参数如下：血清浓度-时间曲线下面积（AUC）= 2,570 +/- 1,650（SD）pg×分钟/毫升，最大浓度（Cmax）= 60 +/- 25（SD）pg/ml，最大时间（Tmax）= 17.5 +/- 6.9（SD）分钟，而肌肉注射ECT（n = 9）的AUC = 9,460 +/- 5,870（SD）pg×分钟/毫升，Cmax = 165 +/- 79（SD）pg/ml，Tmax = 16.1 +/- 4.2（SD）分钟。
