Simple luminescence detectors using a light-emitting diode or a Xe lamp, optical fiber and charge-coupled device, or photomultiplier for determining proteins in capillary electrophoresis: a critical comparison

Analytical Biochemistry
Silvia Casado-TerronesAlberto Fernández-Gutiérrez

Abstract

The performance of two homemade fluorescence-induced capillary electrophoresis detectors, one based on light-emitting diode (LED) as the excitation source and a charge-coupled device (CCD) photodetector and the other based on a commercial luminescence spectrometer (Xe lamp) as the excitation source and a photomultiplier tube as a detector, were compared for the determination of fluorescent proteins R-phycoerythrin and B-phycoerythrin. Both devices use commercially available, reasonably priced optical components that can be used by nonexperts. After fine optimization of several optical and separation parameters in both devices, a zone capillary electrophoresis methodology was achieved with 50mM borate buffer (pH 8.4) and 10mM phytic acid for the determination of two phycobiliproteins. Detection limits of 0.50 and 0.64microg/ml for R-phycoerythrin and B-phycoerythrin, respectively, were achieved by using the LED-induced fluorescence capillary electrophoresis (LED-IF-CE) system, and corresponding detection limits of 2.73 and 2.16microg/ml were achieved by using the Xe lamp-IF-CE system. Analytical performance and other parameters, such as cost and potential to miniaturization, are compared for both devices.

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Citations

Jul 20, 2010·Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy·Yongru BianXiaoming Chen
Jan 15, 2010·Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy·Changqun Cai, Xiaoming Chen
Dec 17, 2008·Electrophoresis·Dan XiaoMartin M F Choi
Dec 7, 2007·Electrophoresis·Vladislav Dolník
Dec 14, 2011·Luminescence : the Journal of Biological and Chemical Luminescence·José A Murillo PulgarínM Nieves Sánchez García

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