Time-resolved detection of hot electron-induced electrochemiluminescence of fluorescein in aqueous solution.
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.
Laboratory of Analytical and Inorganic Chemistry, Helsinki University of Technology, Helsinki, P.O. Box 6100, FIN-02015 HUT, Finland.
Tiina
T
Ylinen
Johanna
J
Suomi
Mika
M
Helin
Timo
T
Ala-Kleme
Sakari
S
Kulmala
D000535:Aluminum / Q000737:chemistry; D004566:Electrodes; D004583:Electrons; D005240:Feasibility Studies; D019793:Fluorescein / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D049449:Luminescence; D008163:Luminescent Measurements / Q000379:methods*; D015394:Molecular Structure; D012996:Solutions / Q000737:chemistry; D013997:Time Factors; D014867:Water / Q000737:chemistry*
D005456:Fluorescent Dyes; D012996:Solutions; D014867:Water; D000535:Aluminum; D019793:Fluorescein