Enzyme immunoassay for 4-hydroxy-2-(4-methylphenyl)benzothiazole.
A specific enzyme immunoassay (EIA) has been developed for 4-hydroxy-2-(4-methylphenyl)benzothiazole (KB-2714) (1), an active metabolite of 4-acetoxy-2-(4-methylphenyl)benzothiazole (KB-2683) (2) which is a promising anti-rheumatic agent. The EIA was based upon antiserum elicited against 5-(2-carboxyphenylazo)-4-hydroxy-2-(4-methylphenyl)benzothiazole (3)-bovine serum albumin conjugate and beta-galactosidase-labelled 5-amino-4-hydroxy-2-(4-methylphenyl)benzothiazole (5). The sensitivity of the EIA was significantly improved by the utilization of a bridge heterologous combination system. An appropriate dose-response curve of EIA for 4-hydroxy-2-(4-methylphenyl)benzothiazole was obtained in the range of 10 pg tube-1-20 ng tube-1. The specificity of EIA proved to be satisfactory in terms of cross-reactivities to 14 benzothiazole-related compounds including glucuronic acid and sulphuric acid conjugates. The proposed method was evaluated to be useful for the determination of 1 in urine and plasma with acceptable recovery and inter- and intra-assay precision.
Pharmaceutical Institute, Tohoku University, Sendai, Japan.
S
S
Miyairi
H
H
Shimada
H
H
Morita
N
N
Awata
J
J
Goto
T
T
Nambara
D000941:Antigens; D052160:Benzothiazoles; D008401:Gas Chromatography-Mass Spectrometry; D006241:Haptens; D006801:Humans; D007106:Immune Sera; D007124:Immunoenzyme Techniques; D012680:Sensitivity and Specificity; D013844:Thiazoles / Q000032:analysis* / Q000097:blood / Q000652:urine; D001616:beta-Galactosidase
D000941:Antigens; D052160:Benzothiazoles; D006241:Haptens; D007106:Immune Sera; D013844:Thiazoles; C083638:4-hydroxy-2-(4-methylphenyl)benzothiazole; D001616:beta-Galactosidase