Abstract
A method is described for construction of a highly sensitive electrochemical biosensor for detection of glutamate. The biosensor is based on covalent immobilization of glutamate oxidase (GluOx) onto polypyrrole nanoparticles and polyaniline composite film (PPyNPs/PANI) electrodeposited onto Au electrode. The enzyme electrode was characterized by cyclic voltammetry (CV), scanning electron microscopy (SEM), X-ray diffraction (XRD), transmission electron microscopy (TEM), Fourier transform infra-red spectroscopy (FTIR) and electrochemical impedance spectroscopy (EIS). The biosensor showed optimum response within 3s at pH 7.5 (0.1M sodium phosphate) and 35°C, when operated at 50mVs−1. It exhibited excellent sensitivity (detection limit as 0.1nM), fast response time and wider linear range (from 0.02 to 400μM). Analytical recovery of added glutamate (5mM and 10mM) was 95.56 and 97%, while within batch and between batch coefficients of variation were 3.2% and 3.35% respectively. The enzyme electrode was used 100 times over a period of 60 days, when stored at 4°C. The biosensor measured glutamate level in food stuff, which correlated well with a standard colorimetric method (r=0.99).
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