Qinghong Jiang’s research while affiliated with Hubei University of Technology and other places

This paper describes a heterogeneous oligonucleotide-hybridization assay based on hot electron-induced electrochemiluminescence (HECL) of a rhodamine label. Thin oxide-film coated aluminum and silicon electrodes were modified with an aminosilane layer and derivatized with short, 15-mer oligonucleotides via diisothiocyanate coupling. Target oligonucleotides were conjugated with tetramethylrhodamine (TAMRA) dye at their amino modified 5′ end and hybridization was detected using HECL of TAMRA. Preliminary results indicate sensitivity down to picomolar level and low nonspecific adsorption. The sensitivity was better on oxide-coated silicon compared to oxide-coated aluminum electrodes and two-base pair mismatched hybrids were successfully discriminated. The experimental results presented here might be useful for the design of disposable electrochemiluminescent DNA biosensors.

Electrogenerated chemiluminescence (ECL) of lucigenin is induced at oxide-coated aluminum electrodes in aqueous solution by cathodic pulse polarization. This ECL can be enhanced by the presence of coreactants such as peroxodisulfalte. The present method is based on the injection of hot electrons into the aqueous electrolyte solution, which probably results in the generation of hydrated electrons as reducing mediators. The successive one-electron redox reactions result in the excited states of lucigenin or its fragmentation products. The method can detect lucigenin over several orders of magnitude of concentration with detection limit below nanomolar concentration level. In addition, the relatively long lifetime of the ECL of lucigenin makes time-resolved detection possible. This study suggests that the derivatives of lucigenin can be utilized as electrochemiluminescent labels in aqueous solution in bioaffinity assays at thin insulating film-coated cathodes. The cathodic ECL reaction mechanisms are discussed.

Synthesis, purification and characterization of [4-ethoxycarbonyl-40-carboxy-2,20-bipyridine]bis(2,2’-bipyridine) ruthenium(II) hexafluorophosphate is described. This complex is shown to be electrochemiluminescent in aqueous solution during cathodic pulse polarization of thin insulating film-coated electrodes. Electrochemiluminescence (ECL) lifetime of the complex was observed to be ca. 40 ms at oxide-coated n-silicon electrodes; thus time-resolved detection is also possible. The ECL emission maximum of this carboxylate derivative is somewhat red-shifted when compared with an unmodified Ru(bpy)3 2+. Because the present complex can be easily covalently coupled with antibodies and oligonucleotides it is usable as an electrochemiluminescent label in various bioaffinity assays. The present chelates also produce strong chemiluminescence during dissolution of metallic magnesium in aqueous solution.

Reactive Al/OH-�(aq) interface was studied and used as a source of chemical energy in the generation of chemiluminescence. The observed extrinsic lyoluminescence emission during dissolution of aluminum in an alkaline Tb(III) or Eu(III) chelate solution was clearly based either on 5D 4 - 7F J radiative transitions of Tb(III) or 5 D 0 - 7 F J transitions of Eu(III). In this process, these chelates were chemically excited via analogous one-electron redox pathways as known from extrinsic lyoluminescence of irradiated, electrolytically- or additively-colored alkali halides, and from hot electron-induced electrochemiluminescence. Calibration curves of Tb(III) chelates, peroxodisulfate and hydroxide ions were linear over several orders of magnitude of concentration. In addition, the method seems to be suitable for relatively rough chemical measurements of the thicknesses of aluminum oxide films free from trapped charges.

A homogeneous immunoassay of T4 was developed using a semiautomatic electrochemiluminometer modified from a commercially available fluorometer. In addition, from the same analyte panel an immunometric immunoassay of TSH at similar disposable oxide-covered aluminum rake electrodes was studied using this instrument both on homogeneous and heterogeneous basis. Detection was based on hot electron-induced cathodic electrochemiluminescence utilizing a commercially available Tb(III) chelate label. The assays were reasonably sensitive and comparison was made with other older methods. Thus, it is possible to develop both non-competitive and competitive immunoassays based on detection of hot electron-induced ECL of the labels.

Hot electrons emitted from thin oxide film-coated heavily doped silicon electrodes by cathodic pulse polarization can induce electrochemiluminescence from luminophores. The intensity of electrochemiluminescence produced at the electrode surface is dependent on the features of thin oxide films formed by thermal oxidation. As a preliminary study, we investigated the effect of thermal oxide growth conditions on the intensity of electrochemiluminescence produced at these electrodes, such as oxidation atmospheres, oxidation temperature, oxidation time and pre-treatment of wafers, using ruthenium(II) tris-(2,2′-bipyridine) chelate as a model luminophore. Optimal oxidation conditions of heavily doped silicon electrodes were obtained for the generation of intense electrochemiluminescence at this kind of silicon electrodes.

Oxide-covered aluminium electrodes were used to demonstrate that aromatic compounds, such as the simple derivatives of benzene, can be electrochemically excited at cathodically pulse-polarized conductor/insulator/electrolyte (C/I/E) tunnel junction electrodes (e.g. oxide-covered aluminium electrodes). The primary cathodic process at these electrodes was a tunnel emission of hot electrons into an aqueous electrolyte solution. Fluorescence (FL) and electrochemiluminescence (ECL) spectra were compared and the dependence of the electrochemiluminescence on the concentrations of benzene, toluene, phenol, p-cresol and aniline were measured and detailed mechanisms for the present electrochemiluminescence are proposed.

Rhodamine B (RhB) exhibits strong cathodic electrogenerated chemiluminescence (ECL) in aqueous solutions during high-amplitude pulse polarization at thin oxide film-coated aluminum electrodes. This method allows the detection of RhB below nanomolar concentration level and provides linear calibration plots spanning over several orders of magnitude of concentration. In addition, a relatively long ECL lifetime of RhB provides a basis for time-resolved detection. Thus, widely used RhB-based labels can also be suggested to be usable as electrochemiluminescent labels in fully aqueous solutions in bioaffinity assays such as in immunoassays and DNA-probing assays. Support was obtained for the chemiluminescence generation mechanism to be essentially the same as that of radiochemiluminescence in aqueous solution.

Double insulating barrier tunnel emission electrodes were fabricated by adding a new pure aluminum layer upon oxidized aluminum electrodes by vacuum evaporation and thermally oxidizing the new aluminum layer in air at room temperature. Resulting Al/Al2O3/Al/Al2O3 electrodes allow the use of various aluminum alloys in the electrode body necessary for hardness or shaping ability of the electrode while obtaining the luminescence properties of pure aluminum oxide. During electrical excitation of luminescent labels by cathodic hot electron injection into aqueous electrolyte solution, the background noise is mainly based on high-field-induced solid-state electroluminescence and F-center luminescence of the outer aluminum oxide film. The more defect states and/or impurity centers the outer oxide film contains, the higher is the background emission intensity. The present electrode fabrication method provides a considerable improvement in signal-to-noise ratio for time-resolved electrochemiluminescence (TR-ECL) measurements when the original native oxide film of the electrode body contains luminescence centers displaying long-lived luminescence. The excellent performance of the present electrodes is demonstrated by extremely low-level detection of Tb(III) chelates, luminol, Pt(II) coproporphyrin and Tb(III) labels in an immunometric immunoassay by time-resolved electrochemiluminescence.

Strong electrogenerated chemiluminescence (ECL) of fluorescein is generated in aqueous solution during cathodic pulse polarization of oxide-covered aluminum electrodes. The present method is based on the injection of hot electrons into the aqueous electrolyte solution, which probably results in the generation of hydrated electrons as reducing mediators. The successive one-electron redox reactions result in fluorescein in its lowest excited singlet state, i.e., the emission spectrum is similar to the corresponding fluorescence emission spectrum. This ECL can be enhanced by added coreactants, e.g., by peroxodisulfate, peroxodiphosphate, hydrogen peroxide and azide ions. The method can detect fluorescein over several orders of magnitude of concentration with detection limit below nanomolar concentration level in the presence of azide ions. The results suggest that the derivatives of fluorescein, such as fluorescein isothiocyanate, can be used as electrochemiluminescent labels in aqueous solution in bioaffinity assays carried out on the surface of thin aluminum oxide film-coated aluminum electrodes. This cathodic ECL is mainly based on the one-electron oxidation of fluorescein by the cathodically produced oxidizing radicals followed by reduction of the formed semioxidized fluorescein by hot or hydrated electrons back to the original oxidation state which finally emits light.