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Optimization of a scanning densitometer for fluorescence detection in high performance thin-layer chromatography
Authors:H. T. Butler  C. F. Poole
Abstract:Conditions for maximizing resolution and signal-to-noise ratios in fluorescence scanning densitometry of separations obtained by high performance thin-layer chromatography are described. Resolution was found to be independent of most instrument parameters with a small decrease observed at wide slit widths. Signal was found to be markedly dependent on experimental parameters such as slit width, slit height, sensitivity setting, and the time constant of electronic filters. For recording chromatograms a slit width of 1.00 mm, a slit height slightly greater than the spot diameter, a low sensitivity setting (50 mV), electronic filtering, and a scan rate of 24 or 48 mm min?1 are commensurate with minimum recording time and maximum sample detectability without incurring a significant decrease in resolution.
Keywords:High performance thin-layer chromatography, HPTLC  Fluorescence quantitation  Scanning densitomery  Resolution  Signal-to-noise ratio
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