Construction of a triglyceride amperometric biosensor based on chitosan-ZnO nanocomposite film

A method is described for construction of a novel amperometric triglyceride (TG) biosensor based on covalent co-immobilization of lipase, glycerol kinase (GK) and glycerol-3-phosphate oxidase (GPO) onto chitosan (CHIT) and zinc oxide nanoparticles (ZnONPs) composite film deposited on the surface of Pt electrode. The enzymes-ZnONPs-CHIT composite was characterized by X-ray diffraction (XRD), scanning electron microscope (SEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The sensor showed optimum response within 6 s at pH 7.5 and temperature of 35 °C. The sensor measures current due to electrons generated at 0.4 V against Ag/AgCl from H2O2, which is produced from triolein by co-immobilized enzymes. A linear relationship was obtained between a wide triolein concentration range (50-650 mg/dl) and current (mA) under optimum conditions. The biosensor showed high sensitivity, low detection limit (20 mg/dl) and good storage stability (half-life of 7 months at 4 °C). The biosensor was unaffected modified by a number of serum substances at their physiological concentrations. The biosensor was evaluated and employed for determination of TG in sera in apparently healthy subjects and persons suffering from hypertriglyceridemia.