1-2hit |
To immobilize cytochrome $c$ (cyt.,$c$) on ITO electrode with keeping its direct electron transfer (DET) activity, 10-carboxydecylphosphonic acid (10-CDPA) self-assembled monolayer (SAM) film was formed on ITO electrode. After 100 times washing process with exchanging phosphate buffer saline solution in the cell to fresh one, extit{in situ} slab optical waveguide (SOWG) absorption spectral measurement proved that about 80% of cyt.,$c$ immobilized on 10-CDPA modified ITO electrode was adsorbed on ITO electrode. Additionally SOWG spectral change of cyt.,$c$ between oxidized and reduced forms was observed with setting the ITO electrode potential at 0.3 and $-$0.3,V vs. Ag/AgCl, respectively showing DET reaction between cyt.,$c$ and ITO electrode occurred. About 30% of a monolayer coverage was estimated from the coulomb amount in the surface area of oxidation and reduction peaks on cyclic voltammetry (CV) data. CV peak current maintained 84% for ITO electrode modified with 10-CDPA SAM film after 60,min continuous scan with 0.1,V/sec from 0.3 and $-$0.3,V vs. Ag/AgCl.
In situ UV-vis. absorption spectra of cytochrome c adsorbed on ITO electrode was observed with slab optical waveguide spectroscopy combining pulse potential step (PPS) between 0.3 and -0.45 V vs. Ag/AgCl. The amount of cytochrome c adsorbed on ITO electrode was estimated from the amount of coulomb of the peaks in cyclic voltammogram to be about a monolayer coverage in this experimental condition. Spectral change between oxidized and reduced cytochrome c by PPS was finished in about 20 msec with phosphate buffer solution. The results strongly proved that SOWG spectroscopy should be effective for in situ observation of ET reaction kinetics of surface adsorbed molecules.