Determination of the bioavailability of intranasal elcatonin in humans: Development of a sandwich transfer enzyme immunoassay for elcatonin

✍ Scribed by Takeyuki Kohno; Noriaki Murasugi; Hiroki Sakurai; Kazuhito Watabe; Hiromichi Nakamuta; Masao Koida; Yohko Sugie; Toshiyo Ogouchi; Tadashi Inoue; Masao Yanaka; Masakatsu Nomura; Akira Yanagawa

• Publisher: John Wiley and Sons
• Year: 1998
• Tongue: English
• Weight: 73 KB
• Volume: 12
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/(sici)1098-2825(1998)12:6<356::aid-jcla5>3.0.co;2-#

✦ Synopsis

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.