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Design and performance of a microchip electrophoresis instrument with sensitive variable-wavelength fluorescence detection

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Belder,  D.
Research Group Belder, Max-Planck-Institut für Kohlenforschung, Max Planck Society;
Research Group Belder, Max-Planck-Institut für Kohlenforschung, Max Planck Society;

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Deege,  A.
Service Department Schulze (GC, HPLC), Max-Planck-Institut für Kohlenforschung, Max Planck Society;
Service Department Schulze (GC, HPLC), Max-Planck-Institut für Kohlenforschung, Max Planck Society;

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Ludwig,  M.
Research Group Belder, Max-Planck-Institut für Kohlenforschung, Max Planck Society;
Research Group Belder, Max-Planck-Institut für Kohlenforschung, Max Planck Society;

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Zitation

Belder, D., Deege, A., Maass, M., & Ludwig, M. (2002). Design and performance of a microchip electrophoresis instrument with sensitive variable-wavelength fluorescence detection. Electrophoresis, 23(14), 2355-2361. doi:10.1002/1522-2683(200207)23:14<2355:AID-ELPS2355>3.0.CO;2-Q.


Zitierlink: http://hdl.handle.net/11858/00-001M-0000-000F-99C8-C
Zusammenfassung
A modular instrument for high-speed microchip electrophoresis (MCE) equipped with a sensitive variable-wavelength fluorescence detection system was developed and evaluated. The experimental setup consists mainly of a lamp-based epifluorescence microscope for variable-wavelength fluorescence detection and imaging and a programmable four-channel bipolar high-voltage source capable of delivering up to +/- 10 W per channel. The optical unit was equipped with a high-sensitivity photomultiplier tube and an adjustable aperture. The system was applied to MCE separations of flurescein isothiocyanate (FITC)- labelled amines utilizing blue light (450-480 nm) for excitation as well as for the separation of rhodamines utilizing excitation light in the green spectral region (531- 560 nm). At optimized conditions baseline separation of four FITC-labelled amines could be obtained in less than 50 s at a detection limit of 460 ppt (1 nM) with a signal-to-noise ratio of 3:1. Three rhodamines could be baseline-separated in less than 6 s at a detection limit of 240 ppt (500 pm). The relative standard deviations of absolute migration times determined in repetitive MCE separations of FITC-labelled amines were below 2.5% (n= 25). By the application of cyclodextrin-modified electrolytes, chiral separation of FITC-labelled amines could be performed in seconds demonstrating the potential of microchip electrophoresis for chiral high-throughput screening.