Detection of NADH and ethanol at a graphite electrode modified with titania sol-gel/Meldola’s Blue/MWCNT/Nafion nanocomposite film

For electrocatalytic determination of NADH, a graphite electrode modified with titania sol-gel/Meldola’s Blue/MWCNT/Nafion nanocomposite was proposed. The composition of the matrix film was optimised in terms of the content of carbon nanotubes and Nafion. Incorporation of a redox mediator, Meldola’s Blue, into the nanocomposite film enabled electrocatalytic determination of NADH at a low potential, −50 mV. For determination of ethanol, alcohol dehydrogenase (ADH) was immobilized into the matrix layer. Experimental conditions affecting the biosensor response were examined, including enzyme loading, temperature of measurement and pH of background electrolyte. Assessments of the analytical characteristics of the biosensor were performed with respect to sensitivity, limit of detection, operational stability, repeatability and reproducibility. The proposed biosensor showed electrocatalytic activity toward oxidation of ethanol with sensitivity of 2.24 µA L mmol−1, linear range from 0.05 to 1.1 mmol L−1, and limit of detection of 25 µmol L−1. The apparent Michaelis-Menten constant was 1.24 mmol L−1, indicating a high biological affinity of ADH/titania sol-gel/Meldola’s Blue/MWCNT/Nafion electrode for ethanol. The developed biosensor was tested in determinations of ethanol content in alcoholic beverages.

Meldola’s Blue — doped titania sol-gel sensor for NADH determination

Titania layers obtained by a sol-gel technique doped with redox mediator, Meldola’s Blue, were employed for construction of a new NADH senor. Optimization of preparation process as well as experimental conditions affecting the response of the sensor were examined. Under optimal conditions NADH could be determined in the wide linear range from 90 to 2300 µM with detection limit 12 µM and a high sensitivity 12.5 nA µM−1. The usefulness of developed sensor was preliminarily checked in determination of NADH forming during enzymatic oxidation of ethanol catalyzed by alcohol dehydrogenase (ADH).