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Time-Resolved Detection of Hot Electron-Induced Electrochemiluminescence of Fluorescein in Aqueous Solution

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Strong electrogenerated chemiluminescence (ECL) of fluorescein is generated during cathodic pulse polarization of oxide-covered aluminum electrodes and the resulting decay of emission is so sluggish that time-resolved detection of fluorescein is feasible. The present ECL in aqueous solution is based on the tunnel emission of hot electrons into the aqueous electrolyte solution, which probably results in the generation of hydrated electrons and hydroxyl radicals acting as redox mediators. The successive one-electron redox steps with the primary radicals result in fluorescein in its lowest excited singlet state. The method allows the detection of fluorescein (or its derivatives containing usable linking groups to biomolecules) over several orders of magnitude of concentration with detection limits well below nanomolar concentration level. The detection limits can still be lowered, e.g., by addition of azide or bromide ions as coreactants. The results suggest that the derivatives of fluorescein, such as fluorescein isothiocyanate (FITC), can be detected by time-resolved measurements and thus be efficiently used as electrochemiluminescent labels in bioaffinity assays.

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Authors and Affiliations

  1. Laboratory of Analytical and Inorganic Chemistry, Helsinki University of Technology, Helsinki, P.O. Box 6100, FIN-02015 HUT, Finland

    Tiina Ylinen, Johanna Suomi, Mika Helin & Sakari Kulmala

  2. Laboratory of Analytical Chemistry, Department of Chemistry, University of Turku, Helsinki, FIN-20014, Finland

    Timo Ala-Kleme

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  1. Tiina Ylinen
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Correspondence to Johanna Suomi.

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Ylinen, T., Suomi, J., Helin, M. et al. Time-Resolved Detection of Hot Electron-Induced Electrochemiluminescence of Fluorescein in Aqueous Solution. J Fluoresc 16, 27–33 (2006). https://doi.org/10.1007/s10895-005-0023-2

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