▎ 摘　　要

NOVELTY - A carbon-based electrode substrate comprises copper metal nanoparticles, carbon nanofibers, and reduced graphene oxide dispersed in a carbonized film. The substrate is copper-carbon nanofibers-reduced graphene oxide/carbon film electro-polymerized with the recognition elements. The carbon nanofibers are grown over the carbon film such that the copper nanoparticles are brought from the inside matrix of the carbon film, barely to the top of the surface. USE - Carbon-based electrode substrate used for electropolymerization of recognition elements as biomarkers to perform electrochemical sensing of biomolecule e.g. glucose, creatinine, cholesterol, and spermine (all claimed) for detecting disease. ADVANTAGE - The carbon-based electrode substrate is manufactured economically and effectively by efficient method in a single reactor without interrupting the process or transferring the samples from one reactor to other, without any structural deformation. DETAILED DESCRIPTION - INDEPENDENT CLAIMS are included for the following: (1) single reactor for preparing substrate which involves treating the polymeric film samples of dried copper nitrate-graphene oxide film in the reactor in which the samples are placed in sandwiched position between meshed plates of the reactor, carbonizing the sample at 900 degrees C for 2 hours in nitrogen atmosphere for converting the polymeric mass to carbon films and dispersed copper nitrate in the films to copper oxide, performing hydrogen reduction by treating the sample with hydrogen gas at 350 degrees C for 2 hours to reduce copper oxide to copper and graphene oxide to reduced graphene, and subjecting the copper-reduced graphene oxide/carbon films under chemical vapor deposition (CVD) to grow the carbon nanofibers on the carbon film samples at the CVD conditions including acetylene flowrate of 30 sccm, temperature 350 degrees C, for 2 minutes; (2) testing/measurement of glucose, which involves electropolymerizing copper-carbon nanofibers-reduced graphene oxide with polyacrylamide by performing cyclic voltammetry in 0.5 M solution of acryl amide and 5 (weight/weight)% N,N'-methylene bis(acrylamide) prepared in 10 mM phosphate buffer solution having pH of 7.4, where the differential pulse voltammetry measurements are carried out at various concentrations of glucose of 20-300 mg/dl prepared in 10 mM phosphate-buffered saline (PBS); (3) testing/measurement of creatinine and cholesterol, which involves electropolymerizing copper-carbon nanofibers-reduced graphene oxide with poly methylene blue and poly methyl orange using 1.0 mM methylene blue and 10 mM methyl orange in 10 mM PBS having pH of 7.4, the poly methylene blue-oxidation and reduction peaks are measured over the creatinine concentration range of 0.5-2.0 mg/dl, and cholesterol measurements are performed over the concentration range of 50-300 mg/dl; and (4) testing/measurement of spermine, which involves electropolymerizing poly cyclodextrin using a 11.36 g/L solution of cyclodextrin in 10 mM PBS having pH of 8.0 where the measurements are performed in concentration of 50-300 mg/l spermine.