Clinical Chemistry
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Clinical Chemistry 30: 1765-1769, 1984;
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Clinical Chemistry, Vol 30, 1765-1769, Copyright © 1984 by American Association for Clinical Chemistry

Single-reagent polarization fluoroimmunoassay for barbiturates in urine

DL Colbert, DS Smith, J Landon and AM Sidki

We describe a rapid polarization fluoroimmunoassay for screening for the presence of barbiturates in urine. The single reagent is prepared by pre-mixing antiserum with a fluorescein-labeled barbiturate derivative as tracer. Use of a mixture of two sheep antisera, raised against different barbiturate immunogens, results in an assay with a broad cross-reactivity spectrum for most common barbiturates. One adds urine to the pre-mixed reagent, incubates the solution for a few minutes at room temperature, and measures the fluorescence polarization. The tracer/antiserum reagent is stable for at least six months at 4 degrees C. Results compare favorably with two other commonly used screening techniques for barbiturates, thin-layer chromatography and the EMIT-stTM (Syva) system. Although designed as a qualitative screen, some drugs can be quantified by means of a standard curve of the relevant barbiturate.




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Copyright © 1984 by the American Association for Clinical Chemistry.