Multilayered graphene (MLG) is an interesting material for electrochemical sensing and biosensing because of its very large 2D electrical conductivity and large surface area. We propose a less toxic, reproducible, and easy method for producing functionalized multilayer graphene from multiwalled carbon nanotubes (MWCNTs) in mass scale using only concentrated H(2)SO(4)/HNO(3). Electron microscopy results show the MLG formation, whereas FTIR and XPS data suggest its carboxylic and hydroxyl-functionalized nature. We utilize this functionalized MLG for the fabrication of a novel amperometric urea biosensor. This biosensor shows linearity of 10-100 mg dL(-1), sensitivity of 5.43 μA mg(-1) dL cm(-2), lower detection limit of 3.9 mg dL(-1), and response time of 10 s. Our results suggest that MLG is a promising material for electrochemical biosensing applications.
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