Biochemie und analytische Biochemie

Abstrakt

Covalent Immobilization of Lactate Oxidase onto Zirconia Coated Silica Nanoparticles/Chitosan Hybrid Film for Amperometric Determination of Lactate

Kusum Dagar and Pundir CS

An improved amperometric L-lactate biosensor was constructed based on covalent immobilization of lactate oxidase (LOx) onto zirconia coated silica nanoparticles (SiO2@ZrONPs)/chitosan (CHIT) hybrid film electrodeposited on the surface of a gold electrode (AuE). The enzyme electrode was characterized by cyclic voltammetry (CV), scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy and electrochemical impedance spectroscopy (EIS), while SiO2@ZrONPs were synthesized by chemical reduction method and characterized by transmission electron microscopy (TEM), UV spectroscopy and X-ray diffraction (XRD). The biosensor showed an optimal response within 3s at pH 7.5 in 0.05M sodium phosphate buffer and 30°C, when operated at 20 mVs-1. The biosensor had a low detection limit of 0.2 nM with a wide working / linear range between 0.1 – 4000 μM. The biosensor was employed for measurement of L-lactic acid level in plasma of apparently healthy and diseased persons. Analytical recovery of added lactic acid (5.0 mM and 10.0 mM)) in plasma was 99% and 96.6% respectively. Within- and between-batch coefficients of variations were 1.79% and 2.89% respectively. There was a good correlation (R2=0.99) between plasma lactate values as measured by standard enzymatic spectrophotometric method and the present biosensor. The enzyme electrode was used 160 times over a period of 120 days, when stored dry at 4°C.