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J.Health Sci., 47(4), 419-423, 2001
A Rapid Enzyme Immunoassay for Cocaine and Benzoylecgonine Using Glucose Oxidase
Mitsune Yamaguchi,a Hiromi Inoue,a Toshiyuki Chikuma,a Junko Itoh,a
Yukiko Makino,b and Hiroshi Hojo*, a
aDepartment of Hygienic Chemistry, Showa Pharmaceutical University, 3-3165 Higashitamagawagakuen, Machida, Tokyo 194-8543, Japan and bKanto-Shin'etsu Regional Narcotic Control Office, Ministry of Health, Labor and Welfare, 2-4-14 Nakameguro, Meguro-ku, Tokyo 153-0061, Japan
A rapid enzyme immunoassay (EIA) for cocaine and its main metabolite, benzoylecgonine (BZE), using glucose oxidase (GOD) was established. Drug concentrations in samples were determined by the inhibition test. BZE-conjugated GOD and samples were added into the wells of a 96-well microtest plate coated with anti-BZE monoclonal antibody. After washings, color development was performed by additions of glucose, horse radish peroxidase (HRP) and 3,3',5,5'-tetramethylbenzidine (TMB), which was a non-mutagenic and non-carcinogenic chromogen. The time required for the immune reaction and the subsequent color development was only 12 min in this EIA. The EIA specifically detected cocaine and BZE at the low detection limit, that is, about 45.4 pg/well for cocaine, and 433 pg/well for BZE, respectively. The values were hardly interfered by urine diluted to 10%. The present assay method was rapid, sensitive, simple, and safe, and was easily carried out anyplace a micro-plate reader was available. Therefore, it might be suitable to screen cocaine and BZE in urine at the scene of abuse.
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