title: A sandwich transfer enzyme immunoassay for salmon calcitonin: determination of the bioavailability of intranasal salmon calcitonin in human.
abstract: A sandwich transfer enzyme immunoassay for salmon calcitonin (SCT) and its usability for the pharmacokinetic study are described. The assay procedure consisted of the reaction of SCT with 2,4-dinitrophenyl biotinyl anti-SCT IgG and anti-SCT Fab'-beta-galactosidase conjugate, trapping onto (anti-2,4-dinitrophenyl bovine serum albumin) IgG-coated polystyrene balls, eluting with epsilon N-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 SCT was 0.05 pg (15 amol)/50 microliters of sample and 1 pg/ml as the concentration. The application of this method has enabled us to directly estimate the bioavailability of SCT dosed intranasally at the therapeutic level (160 IU, 31 micrograms) for its anti-osteoporotic effect as compared to an intramuscular dose (10 IU, 1.9 micrograms). The pharmacokinetic parameters of the intranasal SCT (n = 6) thus estimated were as follows: the area under the blood concentration-time curve (AUC) 9400 +/- 5400 (SD) pg.h/ml, and the mean residence time (MRT) = 42 +/- 14 (SD) min, when the AUC for the intramuscular SCT (n = 3) = 5600 +/- 2000 (SD) pg.h/ml and the MRT = 39 +/- 19 (SD) min.
authors.affiliation: Faculty of Pharmaceutical Sciences, Setsunan University, Osaka, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Kohno
authors.forename: N
authors.initials: N
authors.lastname: Murasugi
authors.forename: H
authors.initials: H
authors.lastname: Sakurai
authors.forename: K
authors.initials: K
authors.lastname: Watabe
authors.forename: H
authors.initials: H
authors.lastname: Nakamuta
authors.forename: M
authors.initials: M
authors.lastname: Koida
authors.forename: Y
authors.initials: Y
authors.lastname: Sugie
authors.forename: M
authors.initials: M
authors.lastname: Nomura
authors.forename: A
authors.initials: A
authors.lastname: Yanagawa
mesh_terms: D019297:2,4-Dinitrophenol; D000281:Administration, Intranasal; D000328:Adult; D001682:Biological Availability; D019921:Biotinylation; D002116:Calcitonin / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics*; D004355:Drug Stability; D006801:Humans; D007124:Immunoenzyme Techniques; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D007700:Kinetics; D008297:Male; D008863:Microspheres; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine; D001616:beta-Galactosidase
chemical_list: D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D012710:Serum Albumin, Bovine; C028815:salmon calcitonin; D002116:Calcitonin; D001616:beta-Galactosidase; D019297:2,4-Dinitrophenol