Abstract
A competitive heterogeneous capillary enzyme immunoassay with electrochemical detection has been developed for phenobarbital in serum. The oxidized primary antibody was attached covalently to the modified interior surface of a microcapillary (22 μl). The competition between analyte phenobarbital and alkaline phosphatase labeled phenobarbital for a limited number of antibody binding sites was complete in 1.5 h. The enzymatic product ( p-aminophenol) from the catalytic conversion of the substrate ( p-aminophenyl phosphate) was detected by amperometric flow injection analysis. The calibration curve for phenobarbital had a detection limit of 30 μg l −1 (2.8 pmoles or 0.65 ng) and a range of 30–3000 μg l −1. The assay could be used to determine the phenobarbital serum concentration in a 4 μl clinical serum sample without pretreatment.
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