Electropolymerized poly(Toluidine Blue)-modified carbon felt for highly sensitive amperometric determination of NADH in flow injection analysis

Poly(pheniothiazine) films were prepared on a porous carbon felt (CF) electrode surface by an electrooxidative polymerization of three phenothiazine derivatives (i.e., Tthionine (TN), Toluidine Blue (TB) and Methylene Blue (MB)) from 0.1 mol/L phosphate buffer solution (pH 7.0). Among the three phenothiazies, the poly(TB) film-modified CF exhibited an excellent electrocatalytic activity for the oxidation of nicotinamide adenine dinucleotide reduced form (NADH) at +0.2 V vs. Ag/AgCl. The poly(TB) film-modified CF was successfully used as working electrode unit of highly sensitive amperometric flow-through detector for NADH. The peak currents (peak heights) were almost unchanged, irrespective of a carrier flow rate ranging from 2.0 to 4.1 mL/min, resulting in the measurement of NADH (ca. 30 samples/hr) at 4.1 mL/min. The peak current responses of NADH showed linear relationship over the concentration range from 1 to 30 mu mol/L (sensitivity: 0.318 mu A/(mu mol/L); correlation coefficient: 0.997). The lower detection limit was found to be 0.3 mu mol/L (S/N = 3).