Inhibitive detection of benzoic acid using a novel phenol biosensor based on polyaniline-polyacrylonitrile composite matrix
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A novel sensitive and stable phenols amperometric biosensor, based on polyaniline-polyacrylonitrile composite matrix, was applied for determination of benzoic acid. The electrochemical biosensor functioning was based on the inhibition effect of benzoic acid on the biocatalytic activity of the polyphenol oxidase (PPO) to its substrate (catechol) in 0.1M phosphate buffer solution (pH 6.5). A potential value of -50 mV versus SCE, and a constant catechol concentration of 20 microM were selective to carry out the amperometric inhibition measurement. The kinetic parameters Michaelis-Menten constant (K(M)(app)) and maximum current (I(max)) in the absence and in the presence of benzoic acid were also evaluated and the possible inhibition mechanism was deduced. The inhibiting action of benzoic acid on the polyphenol oxidase electrode was reversible and of the typical competitive type, with an apparent inhibition constant of 38 microM. This proposed biosensor detected levels of benzoic acid as low as 2x10(-7)M in solution. In addition, the effects of temperature, pH value of solution on the inhibition and the interferences were investigated and discussed herein. Inhibition studies revealed that the proposed electrochemical biosensor was applicable for monitoring benzoic acid in real sample such as milk, yoghurt, sprite and cola.