The performance of a homemade, simple, fluorescence-induced capillary electrophoresis (CE) detector is described here. It is based on LED as excitation source, a bifurcated optical fibre as a waveguide and a CCD as a photodetector. The connection of all the components is fairly easy, even for non-experts. This detector provides a low cost and rapid system for the determination of high-quantum-yield native fluorescence compounds and fluorescence derivatised compounds by CE with direct fluorescence deter mination. R-phycoerythrin and B-phycoerythrin were used as models for native fluorescence compounds and amine labelled with FITC were set as models for the fluorescence derivatised ones. Detection limits of 0.50 and 0.64 μg/mL for R-phycoerythrin and B-phycoerythrin and 1.6 × 10−7 M for FITC-labelled 1,6-diaminohexane were achieved. The homemade LED-IF detector is not expected to displace the LIF-IF one, but offers another possibility and a cheaper way to solve simple analytical problems for determining biomolecules.
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References
Lagu, A.L. (1999) Applications of capillary electrophoresis in biotechnology. Electro-phoresis 20, 3145–3155
Jenkins, M.A., Guerin, M.D. (1997) Capillary electrophoresis procedures for serum protein analysis: comparison with established techniques. J. Chromatogr. B 699, 57–268
Quigley, W.C., Dovichi, N.J. (2004) Capil lary electrophoresis for the analysis of biopol-ymers. Anal. Chem. 76, 4645–4658
Cazes, J. (Ed.) (1997) Analytical instrumen tation handbook, Marcel Dekker, UK
Fernandez-Gutierrez, A., Segura-Carretero, A. (Eds.) (2005) Electroforesis capilar: Aproxi-mación según la técnica de detección, Editorial Universidad de Granada, Granada
Sluszny, C., He, Y., Yeung, E.S. (2005) Light-emitting diode-induced fluorescence detection of native proteins in capillary elec-trophoresis. Electrophoresis 26, 4197–4203
Viskari, P.J., Kinkade, C.S., Colyer, C.L. (2001) Determination of phycobiliproteins by capillary electrophoresis with laser-induced fluorescence detection. Electrophoresis 22, 2327–2335
Viskari, P.J., Colyer, C.L. (2002) Separation and quantification of phycobiliproteins using phytic acid in capillary electrophoresis with laser-induced fluorescence detection. J. Chro-matogr. A 972, 269–276
Johns, C., Macka, M., Haddad, P.R. (2004) Design and performance of a light-emitting diode detector compatible with a commercial capillary electrophoresis instrument. Electro-phoresis 25, 3145–3152
Kuo, J.S., Kuyper, C.L., Allen, P.B., Fiorini, G.S., Chiu, D.T. (2004) High-power blue/ UV light-emitting diodes as excitation sources for sensitive detection. Electrophoresis 25, 3796–3804
Webster, J.R., Burns, M.A., Burke, D.T., Mastrangelo, C.H. (2001) Monolithic capillary electrophoresis device with integrated fluorescence detector. Anal. Chem. 73, 1622–1626
Hillebrand, S., Schoffen, J.L., Mandaji, M. (2002) Performance of an ultraviolet light-emitting diode-induced fluorescence detector in capillary electrophoresis. Electrophoresis 23, 2445–2448
Su, A.K., Lin, C.H. (2003) Determination of riboflavin in urine by capillary electrophoresis–blue light emitting diode-induced fluorescence detection combined with a stacking technique. J. Chromatogr. B 785, 39–46
Su, A.K., Chang, Y.S., Lin, C.H. (2004) Analysis of riboflavin in beer by capillary electrophoresis/blue light emitting diode (LED)-induced fluorescence detection combined with a dynamic pH junction technique. Talanta 64, 970–974
Tsai, C.H., Huang, H.M., Lin, C.H. (2003) Violet light emitting diode-induced fluorescence detection combined with on-line sample concentration techniques for use in capillary electrophoresis. Electrophoresis 24, 3083–3088
Zhao, S., Wang, B., Yuan, H., Xiao, D.J. (2006) Determination of agmatine in biological samples by capillary electrophoresis with optical fiber light-emitting-diode-induced fluorescence detection. J. Chormatogr. A 1123, 138–141
Wang, C., Zhao, S., Yuan, H., Xiao, D. (2006) Determination of excitatory amino acids in biological fluids by capillary elec-trophoresis with optical fiber light-emitting diode induced fluorescence detection. J. Chromatogr. B 833, 129–134
Zhao, S., Wang, B., He, M., Bai, W., Chen, L. (2006) Determination of free D-alanine in the human plasma by capillary electrophore-sis with optical fiber light-emitting diode-induced fluorescence detection. Anal. Chim. Acta 569, 182–187
Yang, B., Guan, Y. (2003) Light-emitting-diode-induced fluorescence detector for capillary electrophoresis using optical fiber with spherical end. Talanta 59, 509–514
Arraez-Roman, D., Fernandez-Sanchez, J.F., Cortacero-Ramirez, S., Segura-Carretero, A. Fernandez-Gutierrez, A. (2006) A simple light-emitted diode-induced fluorescence detector using optical fibers and a charged coupled device for direct and indirect capillary electrophoresis methods. Electrophoresis 27, 1776–1783
Casado-Terrones, S., Cortacero-Ramirez, S., Carrasco-Pancorbo, A., Segura-Carretero, A., Fernández-Gutiérrez, A. (2006) Comparative study between a commercial and a homemade capillary electrophoresis instrument for the simultaneous determination of aminated compounds by induced fluorescence detection. Anal. Bioanal. Chem. 386, 1835–1847
Casado Terrones, S., Fernández Sánchez, J.F., Segura Carretero, A., Fernández Gutiérrez, A. (2007) Simple luminescence detectors using a LED or a Xe lamp, optical fibre and CCD device or photomultiplier for determining proteins in capillary electro-phoresis: a critical comparison. Anal. Bio-chem. 365, 82–90
Arráez-Román, D., Segura-Carretero, A., Cruces-Blanco, C., Fernández-Gutiérrez, A. (2004) Subminute and sensitive determination of the neurotransmitter serotonin in urine by capillary electrophoresis with laser fluorescence detection. Biomed. Chromatogr. 18, 422–426
Acknowledgments
This work was supported by the projects CTQ2005-01914/ BQU of the Ministry of Education and Science and RMN-666 from the Andalusian Government.
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Segura-Carretero, A., Fernández-Sánchez, J.F., Fernández-Gutiérrez, A. (2009). Simple Luminescence Detector for Capillary Electrophoresis. In: Rasooly, A., Herold, K.E. (eds) Biosensors and Biodetection. Methods in Molecular Biology™, vol 503. Humana Press. https://doi.org/10.1007/978-1-60327-567-5_12
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