Single-reagent polarisation fluoroimmunoassay for the cocaine metabolite, benzoylecgonine, in urine.
A polarisation fluoroimmunoassay is described for the detection in urine of benzoylecgonine, the major metabolite of cocaine. The method uses a single reagent comprising sheep anti-benzoylecgonine serum pre-mixed with a fluorescein-labelled benzoylecgonine derivative as tracer. Under the assay conditions the antiserum has equal cross-reactivity with cocaine and the metabolite. The test is performed by the addition of 10 microL of urine to an aliquot of the single reagent, incubation for a few minutes and measurement of fluorescence polarisation. The assay has a positive/negative cut-off level of 1 mg/L benzoylecgonine and results compare favourably with the EMIT-d.a.u.TM (Syva) system.
D000918:Antibody Specificity; D003042:Cocaine / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism* / Q000652:urine; D003429:Cross Reactions; D005454:Fluorescence Polarization; D005455:Fluorescent Antibody Technique; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics
C005618:benzoylecgonine; D003042:Cocaine