The measurement of the anti-epileptic drug phenobarbital from serum samples combining immunoassay and size-exclusion chromatography is presented. The immunoreaction is based on the competitive binding of the analyte (unlabelled phenobarbital) and the fluorescent-labelled phenobarbital to anti-phenobarbital antibodies. Mixing of the reagents and the immunoreaction takes place in a flow system. The products are separated on-line on a short gel chromatographic column and the fluorescence intensity of the marker is measured. The calibration curve shows good linearity in the range 5-80 μg/ml, corresponding to therapeutically relevant serum levels. Intra-day precision values are between 7.32 and 9.48%; the accuracy is between 0.97 and 9.43%. Inter-day precision and accuracy measured on 6 different days fall between 5.38 and 10.05% and -8.27 and -4.97%, respectively. The results obtained with the proposed method show a good correlation with those of other methods (radioimmunoassay and fluorescence polarisation immunoassay) already established in clinical laboratories. (C) 2000 Elsevier Science B.V.
|Number of pages||9|
|Journal||Journal of Chromatography B: Biomedical Sciences and Applications|
|Publication status||Published - Jan 12 2001|
- Fluorescent immunoassay
- Size-exclusion chromatography
ASJC Scopus subject areas