Enzyme immunoassay for phenobarbital.

Publisher Website
Google Scholar
Abstract
A competitive enzyme immunoassay for phenobarbital with alkaline phosphatase as a label is described. p-Aminophenobarbital was used to prepare the immunogen and the alkaline phosphatase conjugate. The IgG fraction prepared from anti-phenobarbital antiserum was bound to Sepharose 4B or adsorbed on polystyrene tubes. The solid phase antibody could not discriminate p-hydroxyphenobarbital from phenobarbital, but had almost no cross-reactivity with metharbital, barbituric acid, thiobarbiturates and other antiepileptic drugs. The ranges of phenobarbital concentration measurable by enzyme immunoassay with Sepharose-IgG (EIA-I) and with polystyrene tube-IgG (EIA-II) were 10 to 100 ng/tube and 100 to 1000ng/tube, respectively. The results obtained by radioimmunoassay correlated well with those obtained by EIA-I (γ=0.995) and EIA-II (γ=0.943). Intra-and inter-assay variations of EIA-II were determined by replicate assays of sera containing 20 μg/ml and 80 μg/ml phenobarbital. The intra-assay coefficients of variation were 3.6% and 5.8%. The inter-assay coefficients of variation were 11.2% and 9.2%. Because of the convenience of the EIA-II procedure, the method is suitable for the routine assay of serum phenobarbital in small clinical laboratories.

This publication has 4 references indexed in Scilit: