▎ 摘　　要

NOVELTY - Method of real-time monitoring of covalent bond formation in solution, involves preparing nano-reduced grained graphene using graphite nanofibers as raw materials, mixing with protein molecule (MP-11) solution, stirring, obtaining MP-11 non-covalent bond modified nano-reduced graphene oxide, washing, diluting, obtaining MP-11/reduced graphene oxide (rGO) solution, adding phosphate buffered saline solution to resultant solution, and recording relation between current and time curve. USE - Method of real-time monitoring of covalent bond formation in solution (claimed). ADVANTAGE - The real-time monitoring of covalent bond formation in solution, is efficiently carried out by simple method with high sensitivity. DETAILED DESCRIPTION - Nano-reduced grained graphene is prepared by mixing 2 g graphite nanofibers, 10 mL concentrated sulfuric acid and 1 g phosphorus pentoxide at 80 degrees C, pre-oxidizing for 30 minutes, cooling to room temperature for 6 hours, diluting using distilled water, suction-filtering, washing to neutral, drying pre-oxidized graphite, mixing with 50 mL concentrated sulfuric acid, gradually adding 6 g potassium permanganate, stirring, controlling temperature at less than 20 degrees C, stirring the mixture in oil bath at 35 degrees C for 2 hours, quenching the reaction by the addition of 280 mL water, further adding 5 mL 30% hydrogen peroxide, dispersing resultant mixture in water, filtering, washing filtered filter cake using dilute hydrochloric acid, drying, preparing 50 mL 0.1 mg/ml oxidized graphene dispersion, adding 50 mg vitamins, and stirring at room temperature for 24 hours. 1 mL MP-11 solution with concentration of 0.186 mg/mL is added to 1 mL 0.05 mg/ml nano-reduced graphene solution, and stirred for 12 hours to obtain MP-11 non-covalent bond modified nano-reduced graphene oxide. The obtained solution is washed to leave excess MP-11 molecules. The resultant mixture is finally diluted to obtain 40 mu M MP-11/rGO solution. A gold electrode with diameter of 12.5 mu m, is immersed in 5 mg/ml Lomant reagent and dimethyl sulfoxide solution, and maintained for 6 hours, to obtain Lomant monolayer modified gold electrode as Lomant/gold electrode. Another gold electrode with diameter of 12.5 mu m is immersed in an alcohol solution of MPA at the concentration of 10 mM for 6 hours to obtain MPA/gold electrode. In presence of Lomant/gold and MPA/Au electrode with diameter of 12.5 mu m as working electrode and silver/silver chloride as reference electrode, 10 mM phosphate buffered saline solution is added to MP-11/graphene, and relation between current and time curve is recorded in the working voltage of -600 mV and data acquisition interval of 50 ms. All the experiments are done at room temperature in the Faraday shield. MP-11/rGO produces covalent bonds on the Lomant/gold electrode surface and Lomant reagent, and resultant current curve is staircase-like which continuously increase in current, while MPA/gold surface is negatively charged. When MP-11/rGO moves to the electrode surface, collision of electrodes in the electrostatic repulsion under the effect of rapid removal is carried out.