Abstract
Peroxyoxalate chemiluminescence was, for the first time, examined by using ternary mixed solutions of water-hydrophilic/hydrophobic organic solvent. Eosin Y as a model fluorescence compound was dissolved with the ternary solutions of water (1.0 mM carbonate buffer, pH 9.0)-acetonitrile-ethyl acetate, water-rich of 15:3:2 volume ratio and organic solvent-rich of 3:8:4 volume ratio, to which bis(2,4,6-trichlorophenyl) oxalate and hydrogen peroxide chemiluminescence reagent were added. The chemiluminescence observed with the ternary solutions, especially the organic solvent-rich solution, showed a larger signal than that observed with the water only solution or water-acetonitrile mixed solution. Chemiluminescence in the presence of twenty types of α-amino acid was similarly examined by using the ternary organic solvent-rich solution. The chemiluminescence of three α-amino acids with fluorescence properties was enhanced with the ternary solution. The data reported here may contribute to development of a new, sensitive peroxyoxalate chemiluminescence detection system.
Highlights
When the mixed solution was fed into a microspace, such as capillary tubes and microchannels on a microchip, annular flow with inner and outer phases was generated under certain conditions; we call this “Tube Radial Distribution Flow” (TRDF)
The tendency of the ternary mixed solutions, especially organic solvent-rich solution, to show large CL based on the peroxyoxalate reaction system was similar to that reported in our previous paper [12], where we examined the influence of the ternary mixed solution on a luminal CL reaction system
The CL of Eosin Y and three α-amino acids were enhanced with the ternary organic solvent-rich solution, compared to the CL observed with the water only and water-acetonitrile mixed solutions
Summary
We reported unique behavior of ternary mixed solutions of water-hydrophilic/hydrophobic organic solvent, such as water-acetonitrile-ethyl acetate mixed solution [5] [6] [7]. The mixing behavior of ternary water-hydrophilic/hydrophobic organic solvent solutions was investigated using a Y-type microchannel. These two mixed solutions, water-acetonitrile and acetonitrile-ethyl acetate, were individually fed into two separate microchannels of a microchip that combined to form a single channel in a Y-type microchannel, affording a ternary mixed solution in the single channel. Denaturation was examined in a ternary mixed solution of water-hydrophilic/ hydrophobic organic solvent using λ-DNA and a plasmid as models. We examined luminal chemiluminescence (CL) behavior in a ternary mixed solvent solution of water-acetonitrile-ethyl acetate, and the mixed solution was found to enhance luminol CL in time and intensity under certain conditions [12]. Bis(2,4,6trichlorophenyl) oxalate (TCPO), Eosin Y, hydrogen peroxide (H2O2), acetonitrile, ethyl acetate, and twenty types of α-amino acids were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan)
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