▎ 摘　　要

NOVELTY - Preparing reduced graphene oxide (RGO)-CMCS-Hemin/PtNPs nano-composite material involves weighing the graphene oxide, placing in distilled water to constant volume to 60 mL, using ultrasonic cell crushing instrument ultrasonic for 1 hour to make it fully dissolve uniformly, preparing 0.1mg/mL of aqueous solution, 10 mg ascorbic acid is added, stirred and reducing for 20 hours to obtain RGO solution. The 20 mg CMCS is added to RGO solution, ultrasonically crushed for 30 minutes, obtaining uniformly mixed RGO-CMCS dispersion. The 10mL of 1 mg/mL of Hemin solution is added in RGO-CMCS solution, crushed for 1 hour to make it fully and uniformly mixed to obtain RGO-CMCS-Hemin dispersion liquid. The 4 ml 0.01mg/mL of sodium chloroplatinate is added in the RGO-CMCS-Hemin dispersion liquid, stirred while adding ascorbic acid 10 mg, stirring 20 hours to obtain RGO-CMCS-Hemin/PtNPs suspension, centrifuged and washed, and dried at 70 degrees C to obtain the RGO-CMCS-Hemin/Pt NPs nano composite material. USE - Method for preparing RGO-CMCS-Hemin/PtNPs nano-composite material. ADVANTAGE - The method has low detection limit and can reach 0.0384 mg/mL. DETAILED DESCRIPTION - An INDEPENDENT CLAIM is included for the method for detecting 1, 5-AG based on nano-composite material, which involves: (A) constructing electrochemical biosensor interface screen printing electrode is firstly soaked in 0.5 mol/LH2SO4 solution for cyclic voltammetry (CV) scanningbefore using, scanning 20 circles in the voltage rangeof -0.4-1.0V, after scanning, washing with water, drying, obtaining the activated SPCE; (B) putting the activated SPCE electrode into 10 mL of 0.01 wt.% chloroauric acid solution, depositing for 120 seconds at -0.4 Volt constant potential, washing and drying to obtain AuNPs/SPCE; (C) soaking the Au NPs/SPCE electrodewith 2.5 wt.% glutaraldehyde for 15 minutes, washing and drying with pH of 7.4 PBS, then dropping 6 L of RGO-CMCS-Hemin/Pt NPs suspension, incubating for 60 minutes, washing by PBS, drying; (D) obtaining RGO-CMCS-Hemin/PtNPs/Au NPs/SPCE; (E) dropping the 3 LPROD to RGO-CMCS-Hemin/Pt NPs/Au NPs/SPCEinterface; (F) incubating for 3h, washing the PROD notcapable of being fixed to the interface, naturally dryingto obtain the PROD/RGO-CMCS-Hemin/Pt NPs/AuNPs/SPCE sensing interface; (G) drawing of5-AG working curve the PROD/RGO-CMCS-Hemin/Pt NPs/Au NPs/SPCE sensing interfacedropping 3 L 1, 5-AG solution, incubating for 30 minutes at 37 degrees C temperature; (H) washing and drying by using PBS solution with pHvalue of 7.4 and distilled water, obtaining 1, 5-AG/PROD/RGO-CMCS-Hemin/Pt NPs/Au NPs/SPCE; (I) putting the 1, 5-AG/PROD/RGO-CMCS-Hemin/PtNPs/Au NPs/SPCE in PBS buffer solution withconcentration of 0.2 mol/L, pH value of 7.4, using DPVscanning of electrochemical working station, recordingthe peak current, the concentration of 5-AG is from 0.1mg/mL to 2 mg/mL, the current response value of thesensor is in linear relationship with the concentrationof 1; 5-AG, where Y is the current peak potential responseintensity, X is 1; 5-AG concentration, linear equationY=4.01372 2.18401X; (J) detecting the 5-AG the normal humanserum sample with 1: 1 of proportion respectively 0.5mg/mL, 1.5 mg/mL, 2.0 mg/mL of 1, 5-AG standard solution fully mixing to prepare mixed solution; (K) dripping 3 Mu L of mixed solution tothe surface of the PROD/RGO-CMCS-Hemin/PtNPs/Au NPs/SPCE to form a working electrode, according to the step 3, the working electrode isplaced in PBS buffer solution, using DPV for scanning; and (L) recording the current value, according to the standard curve, calculating the concentrationof 1, 5-AG in the corresponding actual serum sample.