Fourier processing of liquid chromatograms using flow radioactive detection.
Flow radioactivity counters coupled to liquid chromatography devices cause a systematic bias to the separation by broadening peaks within the radiochromatogram. Such signal smearing may be evaluated on standardization runs with a single peak, using the ratio between Fourier transforms of whole chromatographic data for the measured radioactivity time series (radioactivity channel) and for the concentration time series (optical density channel). This ratio constitutes a kernel suitable to perform the deconvolution of any radiochromatogram performed under similar conditions. Through deconvolution, the signal smearing is removed, reverting to peaks with the same geometry as in the concentration chromatogram: same retention time, peak width, and shape. The deconvolution method in processing radiochromatograms allows an easier interpretation and gives more reliable radioactivity quantification (improved linearity of the measured response). Fourier transformation of the radiochromatogram also allows the removal of transitory events (noise) through the correlation procedure. This method may provide substantial gain in sensitivity, depending upon the residence time of the sample in the counting cell.
Bonnot
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G
Laboratoire de Biologie appliquée, INSA, UA INRA 227, Villeurbanne, France.
Febvay
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G
D002853:Chromatography, Liquid; D005583:Fourier Analysis; D011874:Radiometry