Enzyme-linked immunosorbent assay for TA-2005-glucuronide in human plasma.
A sensitive enzyme-linked immunosorbent assay (ELISA) for TA-2005-glucuronide, a main metabolite of new adrenergic beta-receptor agonist TA-2005, has been investigated without prior deconjugation. Coupling of the hapten with bovine serum albumin (BSA) or beta-D-galactosidase was carried out by the N-hydroxysuccinimide ester method. An anti-TA-2005-glucuronide antiserum was obtained from guinea pig immunized with the hapten-BSA conjugate. The ELISA was based upon a competitive assay in which the separation of bound from free fraction was performed by the double antibody technique using rabbit anti guinea pig immunoglobulin antibody adsorbed to microtiter plates. A satisfactory standard curve for the ELISA of TA-2005-glucuronide was observed in the range of 30 pg-3 ng ml-1 using 25 microliters of human plasma. Inter-day and intra-assay variations were 7.0-17.5% and 1.0-11.7% respectively. The recoveries of TA-2005-glucuronide spiked to plasma samples were 95.5-120% (inter-assay) and 96.0-123.3% (intra-assay). The cross-reactivities of the prepared antiserum with the related compound of TA-2005-glucuronide were quite low though there was a considerable cross-reaction with TA-2005. However, TA-2005-glucuronide could be easily separated from TA-2005 by a simple pretreatment of the plasma sample with a C18 cartridge column. This method was applied to the determination of TA-2005-glucuronide in human plasma samples for the evaluation of the pharmacokinetics of TA-2005. From the results, it was demonstrated that the ELISA developed was useful for the determination of TA-2005-glucuronide in human plasma and that the method was applicable to pharmacokinetic studies in humans.
Matsukawa
M
M
Analytical Research Laboratory, Tanabe Seiyaku Co., Ltd., Osaka, Japan.
Takeda
K
K
Shima
H
H
Tagawa
K
K
Banno
K
K
Sato
T
T
D000284:Administration, Oral; D000318:Adrenergic beta-Agonists / Q000097:blood*; D000662:Amphetamines / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D000818:Animals; D001682:Biological Availability; D002417:Cattle; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D005965:Glucuronates / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D006168:Guinea Pigs; D006801:Humans; D006912:Hydroxyquinolines / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D007106:Immune Sera / Q000737:chemistry / Q000276:immunology; D015363:Quinolones / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D011817:Rabbits; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013388:Succinimides / Q000737:chemistry; D001616:beta-Galactosidase / Q000737:chemistry
C114281:8-hydroxy-5-(1-hydroxy--2-(N-(2-(4-methoxyphenyl)-1-methylethyl)amino)ethyl)carbostyril glucuronide; D000318:Adrenergic beta-Agonists; D000662:Amphetamines; D005965:Glucuronates; D006912:Hydroxyquinolines; D007106:Immune Sera; D015363:Quinolones; D013388:Succinimides; D012710:Serum Albumin, Bovine; C074413:carmoterol; D001616:beta-Galactosidase; C001426:N-hydroxysuccinimide