Cytochrome C based two-channel ratio type hydrogen peroxide biosensor, and preparation method and application thereof
A biosensor, hydrogen peroxide technology, applied in the direction of material analysis, instruments, and scientific instruments by electromagnetic means, can solve the problems of difficult acquisition, inaccurate results, poor stability, etc., and achieve good electrocatalytic effect and conduction efficiency. Good and stable effect
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Embodiment 1
[0036] Example 1 Preparation of dual-channel ratiometric hydrogen peroxide biosensor based on cytochrome C
[0037] Before the experiment, the gold electrode was polished on metallographic sandpaper, ultrasonically cleaned with 1:1 ethanol, 1:1 nitric acid and deionized water in sequence, and dried for later use.
[0038] Step 1. Soak the Au electrode (electrode ①) into a 1mmol / L MPA solution, take it out after 10 minutes, and wash it with deionized water to obtain an Au / MPA electrode;
[0039] Step 2. Mix and react EDC (5mmol / L) and NHS (2mmol / L) for 5 hours, adjust the pH to neutral, add Cyt.C solution with a concentration of 5mg / mL, and soak the Au / MPA electrode into the mixture After reacting at room temperature for 1 hour, take it out, wash off the EDC-NHS residue, dry it for later use, and obtain an Au / MPA / Cyt.C electrode (such as figure 2 shown);
[0040] Step 3. Soak the other Au electrode in the FcHT-ethanol solution with a volume ratio of 1:2000, store it away from ...
Embodiment 2
[0041] Modified Characterization, Electrochemical Performance and Stability of Example 2 Au / MPA / Cyt.C Electrode and Au / FcHT Electrode
[0042] (1) Modification and characterization of Au / MPA / Cyt.C electrode and Au / FcHT electrode
[0043] The Au / MPA / Cyt.C electrode and Au / FcHT of Example 1 were used as the working electrode and the reference channel electrode, the Ag / AgCl electrode was used as the reference electrode, and the Pt wire was used as the counter electrode to construct a three-electrode system. The result is as Figure 3A with Figure 3B shown.
[0044] Figure 3A The results showed that only charging current was observed on the Au electrode and Au / MPA electrode, but a pair of clear redox peaks could be observed on the Au / MPA / Cyt.C electrode, indicating that Cyt.C had been successfully modified on the electrode;
[0045] Figure 3B The results showed that only charging current was observed on the Au electrode, while a pair of clear redox peaks could be observed ...
Embodiment 3
[0050] Embodiment 3 electrocatalytic performance
[0051] Such as figure 1 As shown, the prepared Au / MPA / Cyt.C and Au / FcHT electrodes were respectively connected to the CHI 760E electrochemical workstation, and a four-electrode system was formed with the Ag / AgCl electrode and the platinum wire counter electrode, and the differential pulse voltammetry was selected. (DPV) was measured. The initial potential of Au / MPA / Cyt.C is 0.4V, the final potential is -0.1V, the initial potential of Au / FcHT is 0.7V, the final potential is 0.1V, the potential increment is 0.004V, and the test solution is 0.1mol / L PBS solution, pH=7.0. The result is as Image 6 shown.
[0052] Depend on Image 6 It can be seen that with H 2 o 2 With the increase of the concentration, the reduction peak current of Cyt.C increased continuously; while the peak current of the reference peak FcHT remained basically the same. Correlation coefficient R of the ratio of Cyt.C response peak current to FcHT refere...
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