Nanochemistry and biotechnology

Development of Enzyme Amperometric Biosensor Based on Poly(meta-phenylenediamine) Film for Determination of Lactose 

V.M. Pyeshkova1,2, O.Y. Dudchenko2, O.V. Soldatkinа1,2, T. Seker3, B. Akata Kurc3, S.V. Dzyadevycha1,2

1 Institute of Molecular Biology and Genetics of NAS of Ukraine, Department of Biomolecular Electronics, Zabolotnogo Street 150, 03680 Kyiv, Ukraine;
E-mail: victoriya.p@gmail.com
2 Institute of High Technologies of Taras Shevchenko National University of Kyiv, Hlushkova Avenue, 4g, 03022, Kyiv, Ukraine;
3Middle East Technical University, Central Laboratory, 06531, Turkey, Ankara;

New amperometric enzyme biosensor based on platinum disk electrodes modified by nanosized polyphenylenediamine (PPD) film for determination of lactose was developed in this work. The PPD was used to form a semipermeable film on the electrode surface to prevent influence of interfering substances. Due to the presence of certain pores in such film it was able to pass compounds with low molecular weight (e.g. hydrogen peroxide) and to retain larger molecules (e.g. ascorbic acid). The bioselective membrane of biosensor was obtained by immobilization of three enzymes (β-Galactosidase, Mutarotase and Glucose Oxidase) on the surface of amperometric transducer (platinum electrode modified by PPD film).
The analytical characteristics of the developed biosensor were studied. The linear range of lactose determination was from 0.005 mM to 1.25 mM. The time of lactose determination was around 30 seconds. The biosensor showed high sensitivity, operational stability, selectivity and signal reproducibility. The developed biosensor was applied for lactose determination in milk samples. It was shown high correlation between results obtained with lactose biosensor and HPLC. Thus, the developed enzyme biosensor based on nanosized PPD film is suited for rapid, inexpensive, sensitive, selective and simple determination of lactose in milk samples.