Fabrication of a Novel Optical Glucose Biosensor Using Copper(II) Neocuproine as a Chromogenic Oxidant and Glucose Dehydrogenase-Immobilized Magnetite Nanoparticles.

This study describes a novel optical glucose biosensor based on a colorimetric reaction between reduced nicotinamide adenine dinucleotide (NADH) and a copper(II) neocuproine complex ([Cu(Nc) 2 ] 2+ ) as a chromogenic oxidant. An enzymatic reaction takes place between glucose and glucose dehydrogenase (GDH)-chitosan (CS) immobilized on silanized magnetite nanoparticles (CS@SiO 2 @Fe 3 O 4 ) in the presence of coenzyme NAD + . The oxidation of glucose to gluconolactone via the immobilized enzyme is coupled with the reduction of NAD + to NADH at the same time. After the separation of GDH-immobilized SiO 2 @Fe 3 O 4 with a magnet, the enzymatically produced NADH chemically reduces the chromogenic oxidant cupric neocuproine to the cuprous chelate. Thus, the glucose biosensor is fabricated based on the measurement of the absorbance of the formed yellow-orange complex ([Cu(Nc) 2 ] + ) at 450 nm. The obtained results show that the colorimetric biosensor has a wide linear response range for glucose, between 1.0 and 150.0 μM under optimized conditions. The limit of detection and limit of quantification were found to be 0.31 and 1.02 μM, respectively. The selectivity properties of the fabricated biosensor were tested with various interfering species. This biosensor was applied to various samples, and the obtained results suggest that the fabricated optical biosensor can be successfully used for the selective and sensitive determination of glucose in real samples.