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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.
Anal Biochem. 2007 Jun 01; 365(1):82-90.AB

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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Authors+Show Affiliations

Casado-Terrones S
Department of Analytical Chemistry, Faculty of Sciences, University of Granada, E-18071 Granada, Spain.
Fernández-Sánchez JF
No affiliation info available
Segura-Carretero A
No affiliation info available
Fernández-Gutiérrez A
No affiliation info available

MeSH

Electrophoresis, CapillaryEquipment DesignFiber Optic TechnologyLuminescenceOptical FibersPhycobiliproteinsPhycoerythrinProteinsSensitivity and SpecificityXenon

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17391633

Citation

Casado-Terrones, Silvia, et al. "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, vol. 365, no. 1, 2007, pp. 82-90.
Casado-Terrones S, Fernández-Sánchez JF, Segura-Carretero A, et al. 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. Anal Biochem. 2007;365(1):82-90.
Casado-Terrones, S., Fernández-Sánchez, J. F., Segura-Carretero, A., & Fernández-Gutiérrez, A. (2007). 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, 365(1), 82-90.
Casado-Terrones S, et al. 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. Anal Biochem. 2007 Jun 1;365(1):82-90. PubMed PMID: 17391633.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Casado-Terrones,Silvia, AU - Fernández-Sánchez,Jorge F, AU - Segura-Carretero,Antonio, AU - Fernández-Gutiérrez,Alberto, Y1 - 2007/02/23/ PY - 2007/01/15/received PY - 2007/02/07/revised PY - 2007/02/09/accepted PY - 2007/3/30/pubmed PY - 2007/8/19/medline PY - 2007/3/30/entrez SP - 82 EP - 90 JF - Analytical biochemistry JO - Anal Biochem VL - 365 IS - 1 N2 - 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. SN - 0003-2697 UR - https://www.unboundmedicine.com/medline/citation/17391633/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_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-2697(07)00100-5 DB - PRIME DP - Unbound Medicine ER -