Feasibility study of a drug immunoassay based on micellar electrokinetic capillary chromatography with laser induced fluorescence detection: determination of theophylline in serum.
This paper presents principle and first results of a novel competitive binding immunoassay for monitoring of theophylline in human serum. The assay is based upon short time incubation of a mixture of antiserum, containing the antibody raised against theophylline, fluorescein labelled theophylline (tracer) and serum prior to injection of a few nanoliter of this mixture onto a fused-silica capillary for subsequent separation and analysis of free tracer and the antibody-tracer-complex by micellar electrokinetic capillary chromatography with laser induced fluorescence detection. Quantitation based upon multi-level calibration using the height of the peak produced by the free tracer is shown to provide theophylline serum levels which are in agreement with those obtained by a commercial fluorescence polarization immunoassay and with those determined by micellar elektrokinetic capillary chromatography with direct serum injection and on-column UV absorption detection.
Steinmann
L
L
Department of Clinical Pharmacology, University of Bern, Switzerland.
Caslavska
J
J
Thormann
W
W
D001667:Binding, Competitive; D002197:Capillary Action; D002845:Chromatography / Q000379:methods*; D016903:Drug Monitoring; D004563:Electrochemistry; D005240:Feasibility Studies; D016231:Fluorescence Polarization Immunoassay / Q000379:methods*; D006801:Humans; D007700:Kinetics; D007834:Lasers; D008823:Micelles; D012016:Reference Values; D013806:Theophylline / Q000097:blood*
D008823:Micelles; D013806:Theophylline