Glucose biosensor based on entrapment of glucose oxidase and myoglobin in silica gel by the sol-gel method

Abstract

A spectrophotometric method is presented to determine glucose employing the sol-gel technique. Myoglobin (Mb) and glucose oxidase are encapsulated in a transparent and porous silica glass. The produced gel (xerogel) is then immersed in water where increments of glucose are added to the solution with stirring; glucose diffuses into the sol-gel glass pores and a series of reactions take place. Glucose is first oxidized by glucose oxidase and oxygen to gluconate and hydrogen peroxide is generated. The liberated hydrogen peroxide oxidizes the Mb heme (Fe²⁺ into Fe³⁺). The higher is the glucose concentration added, the more is the H₂O₂ generated, and the more is the Mb oxidation (Fe²⁺ to Fe³⁺) and as a result the higher is the absorbance at 400 nm (negative peak, lower absorbance value). All measurements are performed at this wavelength (400 nm), the negative peak obtained by subtracting the absorption spectra of Mb before and after oxidation. Measuring the slope of the absorbance decay versus time at 400 nm monitors increments of added glucose. Each glucose concentration has an accompanying unique decay curve with a unique slope. The higher is the glucose concentration; the steeper is the decay curve (higher slope value). The calibration curve was linear up to 40 mM.