CN114852995B - Electrocatalytic application of horseradish peroxidase sensor constructed by black phosphorus-based composite material - Google Patents

Electrocatalytic application of horseradish peroxidase sensor constructed by black phosphorus-based composite material Download PDF

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CN114852995B
CN114852995B CN202210331413.2A CN202210331413A CN114852995B CN 114852995 B CN114852995 B CN 114852995B CN 202210331413 A CN202210331413 A CN 202210331413A CN 114852995 B CN114852995 B CN 114852995B
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swcnts
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CN114852995A (en
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孙伟
李小青
姚昱岑
王宝丽
艾益静
邵波
张程
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Hainan Normal University
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Abstract

The invention relates to a preparation method of a thin-layer black phosphane/single-wall carbon nano tube composite material and an electrocatalytic application for constructing a horseradish peroxidase electrochemical sensor. A nanocomposite material is prepared by adopting a liquid phase stripping method and a physical mixing method and is used for modifying horseradish peroxidase to construct an enzyme electrochemical sensor, and is also used for electrocatalytic application of trichloroacetic acid, sodium nitrite and hydrogen peroxide. The method mainly comprises the following steps: preparing a thin layer of black phosphane, compounding the thin layer of black phosphane with a single-wall carbon nano tube, preparing a thin layer of black phosphane/single-wall carbon nano tube composite material, and modifying the composite material to the surface of an ionic liquid electrode to prepare an enzyme electrochemical sensor with a sandwich structure and electrocatalytic application of the enzyme electrochemical sensor. The composite material prepared by the invention has larger effective area, special morphology and stronger conductivity, can improve the stability and the dispersibility of the black phosphazene, and has simple preparation process, sensitive detection and good measurement effect.

Description

Electrocatalytic application of horseradish peroxidase sensor constructed by black phosphorus-based composite material
Technical Field
The invention belongs to the field of preparation methods and application of electrochemical sensors, and relates to preparation of a Black Phosphazene (BPNs)/single-walled carbon nanotubes (SWCNTs) composite material and electrocatalytic application of the composite material in construction of a horseradish peroxidase (HRP) electrochemical sensor.
Background
Black Phosphorus (BP) is an allotrope of phosphorus, commonly called phosphazene, and two adjacent BP layers are overlapped together through van der waals force, so that the black phosphorus has a plurality of excellent performances, such as direct band gap with controllable layer number, higher carrier mobility, obvious anisotropy and better biocompatibility, and has wide application prospects in the fields of photoelectrochemistry, biomedicine, solar cells, lithium ion cells/sodium ion cells, transistors, electronic components and the like. With the intensive research on BP materials, some defects of BP are also exposed, the stability is poor, and BP exposed to air or placed in an aqueous solution in the presence of light can be degraded to generate nontoxic PxOy oxide. Fortunately, by performing effective surface functionalization on BP, further degradation of BP can be avoided, resulting in BP with good stability and dispersibility.
The thin BPNs have more outstanding advantages such as strong conductivity, good biocompatibility and the like. On the surface of BPNs, SWCNTs material is modified, which can play a role in delaying the degradation of BPNs. Meanwhile, the BPNs and SWCNTs are synthesized into the BPNs-SWCNTs composite material by adopting a physical mixing method, and the composite material has the characteristics of SWCNTs, such as good biocompatibility, rapid electron transfer, higher surface activity and the like. So that the SWCNTs and the BPNs are compounded to realize the synergistic effect of the SWCNTs and the BPNs. On one hand, SWCNTs play a double role of delaying the degradation of BPNs, and on the other hand, the conductivity of the composite material is improved, so that a wider application prospect is provided for constructing a novel biosensor.
Disclosure of Invention
In view of the above, the invention provides a preparation method of a thin-layer Black Phosphazene (BPNs)/single-wall carbon nanotubes (SWCNTs) composite material and an electrocatalytic application of the composite material for constructing a horseradish peroxidase (HRP) electrochemical sensor. The invention provides synthesis of a BPNs-SWCNTs nanocomposite, the thin-layer functionalized BPNs-SWCNTs nanocomposite has a sheet structure of BPNs and a tubular structure of SWCNTs, and the SWCNTs and the BPNs are intertwined, so that the composite has stronger conductivity and can improve the dispersibility and stability of the thin-layer BPNs.
Preferably, the method comprises the steps of,the effective surface area of the thin layer BPNs-SWCNTs nanocomposite is 0.182 and 0.182cm 2 ；
Preferably, the two BPNs-SWCNTs composites are cross-linked and entangled with each other and the SWCNTs are capable of adhering to the surface of the thin layer BPNs.
The invention provides a preparation method of the composite material and application of an electrochemical enzyme sensor, which comprise the following steps: (1) Mixing BPNPs and NMP in ice bath, performing ultrasonic treatment for 12 hours, centrifuging to prepare a thin-layer BPNs dispersion liquid, and mixing the thin-layer BPNs dispersion liquid and SWCNTs for ultrasonic treatment for 4 hours to prepare a thin-layer BPNs-SWCNTs mixed liquid; (2) With graphite powder and ionic liquid (HPPF) 6 ) The mass ratio is 2:1, mixing, grinding, loading into a glass tube inserted with copper wires to prepare CILE, taking the CILE as a substrate electrode, and polishing the surface of the electrode before each use; (3) Based on the step 2, the CILE and the composite material are put into a glove box, the glove box is vacuumized firstly, and then N is introduced 2 The method comprises the steps of carrying out a first treatment on the surface of the (4) Firstly, dripping BPNs-SWCNTs on the surface of a CILE, and airing at room temperature to obtain a BPNs-SWCNTs/CILE modified electrode; dripping HRP on the BPNs-SWCNTs/CILE electrode, and airing at room temperature to obtain an HRP/BPNs-SWCNTs/CILE modified electrode; and (5) carrying out electrochemical test on the electrode construction three-electrode system obtained in the step (4).
Preferably, the thin layer BPNs-SWCNTs nanocomposite is used in an amount of 10 μl, and the volume ratio of BPNs to SWCNTs is 1: 1. The BPNs are of a lamellar structure, the concentration of HRP is 15.0mg/mL, and the electrode preparation steps (3) and (4) are all filled with N 2 Is completed in a glove box.
Preferably, the preparation of the composite nano material adopts a liquid-phase ultrasonic stripping method and a physical mixing method, and the liquid-phase ultrasonic stripping method prepares the nano composite material by a thin-layer BPNs (binary phase-change copolymers) and the physical mixing method.
Preferably, the preparation of the modified electrode is carried out with the dosage of immobilized enzyme of 10.0 mu L of 15.0mg/mL and naturally airing.
The invention providesThe preparation of the thin-layer Black Phosphazene (BPNs)/single-wall carbon nano tubes (SWCNTs) composite material and the electrocatalytic application for constructing the horseradish peroxidase (HRP) electrochemical sensor are disclosed in the technical scheme. The thin-layer BPNs-SWCNTs nanocomposite provided by the invention has the characteristics of good biocompatibility, rapid electron transfer, higher surface activity, smaller thickness of the BPNs, larger specific surface area and more active centers of the SWCNTs. So the combination of SWCNTs and BPNs can realize the synergy of the SWCNTs and the BPNs. Meanwhile, the conductivity of the composite material can be improved, and the environmental stability and dispersibility of the BPNs are enhanced. The results of the examples show that the BPNs-SWCNTs composite modified electrode provided by the invention is 1.0 mmol/L K 3 [Fe(CN) 6 ]And 0.5mol/L KCl mixed solution, and performing cyclic voltammetry test, wherein the effective area is 0.182cm 2 1.44 times larger than the CILE.
Drawings
FIG. 1 is a scanning electron microscope and a transmission electron microscope image of BPNPs, SWCNTs and thin layer BPNs-SWCNTs nanocomposite prepared in example 1.
FIG. 2 is an X-ray diffraction pattern of the thin-layer BPNs-SWCNTs nanocomposite prepared in example 1.
FIG. 3 is a graph showing the UV-visible absorption spectrum of the HRP of example 1 and the prepared BPNs-SWCNTs-HRP.
FIG. 4 is an infrared spectrum of the HRP and BPNs-SWCNTs composite prepared in example 1.
FIG. 5 shows that the different modified electrodes prepared in test example 1 were at 1.0 mmol/L K 3 [Fe(CN) 6 ]And cyclic voltammograms at different sweep rates in 0.5mol/L KCl mixed electrolyte.
FIG. 6 shows that the BPNs-SWCNTs/CILE modified electrode prepared in test example 1 was used at 1.0 mmol/L K 3 [Fe(CN) 6 ]And a scan rate profile in 0.5mol/L KCl mixed electrolyte.
FIG. 7 shows the redox peak current and v of the BPNs-SWCNTs/CILE modified electrode prepared in test example 1 at different scanning speeds 1/2 Is a linear relationship of (c).
FIG. 8 is a cyclic voltammogram of a different modified electrode prepared in test example 2 in 0.1mmol/L Phosphate Buffered Saline (PBS).
FIG. 9 is a graph showing the scanning speed of Nafion/HRP/BPNs-SWCNTs/CILE modified electrodes prepared in test example 2 in 0.1mmol/L PBS at different pH values.
FIG. 10 is a graph of the Nafion/HRP/BPNs-SWCNTs/CILE modified electrode catalytic TCV prepared in test example 3.
FIG. 11 shows Nafion/HRP/BPNs-SWCNTs/CILE modified electrode catalytic NaNO prepared in test example 3 2 Is a curve of (2).
FIG. 12 shows a Nafion/HRP/BPNs-SWCNTs/CILE modified electrode catalyst H prepared in test example 3 2 O 2 Is a curve of (2).
Detailed Description
The invention provides a preparation method of a thin-layer Black Phosphazene (BPNs)/single-wall carbon nano tubes (SWCNTs) composite material and an electrocatalytic application of the composite material in construction of a horseradish peroxidase (HRP) electrochemical sensor. The invention provides an electrochemical enzyme sensor which has simple preparation method, easy operation and low cost and is used for measuring TCA and NaNO 2 And H 2 O 2 The electrochemical enzyme sensor has wide electrochemical detection range and low detection limit.
The effective area of an electrochemical sensing platform (BPNs-SWCNTs/CILE) prepared by the thin-layer BPNs-SWCNTs nano composite material provided by the invention is 0.182cm 2 。
Preferably, the electrochemical enzyme sensor is prepared by filling N 2 Is carried out in a glove box.
In the invention, the preparation method of the thin-layer BPNs-SWCNTs nanocomposite and the preparation method of the BPNs-SWCNTs/CILE obtained by modifying electrodes preferably comprise the following steps:
firstly, a liquid phase stripping method and a physical mixing method are adopted to prepare the thin-layer BPNs-SWCNTs nano composite material. Then the CILE and the composite nano material are put into a glove box, the glove box is vacuumized, and then N is introduced 2 And then, dripping the thin BPNs-SWCNTs on the surface of the CILE, and naturally airing to obtain the BPNs-SWCNTs/CILE modified electrode.
In the present invention, the BPNs dispersion is a thin layer BPNs dispersion; the BPNPs are preferably pretreated in the present invention, and in the present invention, the pretreatment preferably includes: the BPNPs solution is sonicated, preferably for a period of 8-12 h, more preferably 12 h, at 10000rmp and 12000rmp for 20 minutes, respectively, and then sonicated with SWCNTs directly mixed for 4 hours.
The thin-layer BPNs-SWCNTs/CILE modified electrode is preferably subjected to post-treatment, and in the method, the post-treatment is preferably naturally dried in a glove box at room temperature. After the BPNs-SWCNTs/CILE modified electrode is obtained, the HRP is continuously dripped, and after natural airing, the HRP/BPNs-SWCNTs/CILE modified electrode is obtained, and the modified electrode has the characteristics of large specific surface area and high conductivity.
In the present invention, the preferable concentration of HRP is 15.0. 15.0mg/mL, and the preferable volume is 10.0. Mu.L.
Finally, dripping Nafion on the HRP/BPNs-SWCNTs/CILE modified electrode to obtain Nafion/HRP/BPNs-SWCNTs/CILE; in the invention, the modified electrode is refrigerated in a refrigerator at 4 ℃ when not in use, and has no special requirement on the preservation time, and the preservation temperature is under natural conditions.
In the present invention, the HRP volume is preferably 10.0. Mu.L. After the modified electrode is obtained, the modified electrode is preferably dried in a glove box at room temperature, so that the Nafion/HRP/BPNs-SWCNTs/CILE working electrode with a sandwich structure is obtained. In the present invention, the reference electrode is preferably a saturated calomel electrode, and the counter electrode is preferably a platinum wire electrode; the invention has no special requirements on the thickness of the dripping and the specific implementation process of the coating, and the conventional dripping thickness and operation well known to the person skilled in the art are adopted; in the invention, the drying time is not particularly required, and the drying is finished.
The technical solutions provided by the present invention are described in detail below in conjunction with examples for further illustrating the present invention, but they should not be construed as limiting the scope of the present invention.
Example 1
Firstly, mixing BPNPs and NMP in an ice bath, performing ultrasonic treatment for 12 hours, and then centrifuging 10000rmp and 12000rmp for 20 minutes respectively to prepare a thin-layer BPNs dispersion liquid; mixing the BPNs dispersion liquid and SWCNTs by a physical mixing method, and performing ultrasonic treatment for 4 hoursPreparing a thin-layer BPNs-SWCNTs nanocomposite; polishing the surface of the CILE electrode, putting the CILE electrode and the composite material into a glove box, vacuumizing the glove box, and then introducing N 2 The method comprises the steps of carrying out a first treatment on the surface of the Firstly, the BPNs-SWCNTs nano composite material is dripped on the surface of the CILE, and after natural airing, the BPNs-SWCNTs/CILE modified electrode is obtained. Other preparation methods of the modified electrodes SWCNTs/CILE and the BPNs/CILE all adopt a coating method, and the BPNs/CILE and the BPNs-SWCNTs/CILE electrodes are not stored in a refrigerator at 4 ℃ for refrigeration.
FIG. 1 is a scanning electron microscope and a transmission electron microscope of different materials, wherein A in FIG. 1 is a scanning electron microscope photograph of a multi-layer BPNPs with a scale of 1 μm, and the BPNPs can be seen from the drawing to show a multi-layer sheet structure; b in FIG. 1 is a scanning electron microscope image of SWCNTs on a scale 100 nm, from which it can be seen that SWCNTs have a tubular structure; c in FIG. 1 is a scanning electron microscope image of the BPNs-SWCNTs composite material when the scale is 100 nm, and the two materials are mutually wound and adhered together; in FIG. 1, D is a transmission electron micrograph of BPNs at a scale of 1 μm, and it can be seen that BPNs are lamellar.
FIG. 2 is an XPD diagram of the prepared thin-layer BPNs-SWCNTs nanocomposite, and the crystallographic characteristics of the synthesized composite are detected by XRD, so that the composite has characteristic diffraction peaks of BPNs and XRD peaks of SWCNTs, and the composite is successful.
FIG. 3 is a graph of UV-visible absorption spectra of HRP and HRP in the preparation of BPNs-SWCNTs composite, and it is apparent from FIG. 3 that Soret absorption bands of HRP in water (curve a) and HRP in BPNs-SWCNTs mixed solution (curve b) both appear at 403.2 nm, indicating that no denaturation of HRP occurs in BPNs-SWCNTs composite, and that BPNs-SWCNTs have good biocompatibility.
FIG. 4 is an infrared spectrum of HRP and HRP in the preparation of composites, and it is apparent from FIG. 4 that the infrared absorption bands of amide I and amide II in BPNs-SWCNTs composites (curve a) and HRP are located at 1700-1600 cm, respectively -1 And 1600-1500 cm -1 The positions of the two were substantially identical, indicating that HRP maintained its original conformation in the BPNs-SWCNTs.
Test example 1
SWCNTs/CILE, BPNs/CILE and BPNs-SWCNTs/CILE electrodes prepared in example 1 were used as working electrodes, saturated calomel electrode was used as reference electrode, platinum wire was used as counter electrode, and the three electrodes were placed at 1.0 mmol/L K 3 [Fe(CN) 6 ]And performing electrochemical performance test in 0.5mol/L KCl mixed electrolyte by adopting a cyclic voltammetry method.
FIG. 5 shows that the BPNs-SWCNTs/CILE modified electrode prepared in test example 1 was used at 1mmol/L K 3 [Fe(CN) 6 ]And a cyclic voltammogram of 100 mV/s in 0.5mol/L KCl mixed electrolyte, as shown in FIG. 5, with a pair of aligned reversible redox peaks on CILE (curve a), a reduction peak current of 38.00 μA for Ipc and an oxidation peak current of 34.39 μA for Ipa. On SWCNTs/CILE (curve b), the redox peak current increases, indicating that SWCNTs have good conductivity. The increase in redox peak current at BPNs/CILE (curve c) was further increased, indicating that BPNs promote [ Fe (CN) more than SWCNTs 6 ] 3-/4- Electron transfer rate. The redox peak current is obviously maximum on BPNs-SWCNTs/CILE (curve d), the reduction peak current and the oxidation peak current are 81.43 mu A and 75.82 mu A respectively, and are increased by 2.14 times and 2.20 times respectively compared with CILE, because of the synergistic effect of BPNPs-SWCNTs, the [ Fe (CN) is accelerated 6 ] 3-/4- The electron transfer rate with the electrode surface improves the electrochemical response signal.
FIG. 6 shows that the BPNs-SWCNTs/CILE modified electrode prepared in test example 1 was used at 1.0 mmol/L K 3 [Fe(CN) 6 ]And cyclic voltammograms of 0.5mol/L KCl mixed electrolyte at different sweeping speeds, the oxidation-reduction peak potential is respectively shifted to positive direction and negative direction along with the increase of the sweeping speed, and the oxidation-reduction peak current is gradually increased along with the increase of the sweeping speed.
FIG. 7 shows the redox peak current and v of the prepared BPNs-SWCNTs/CILE modified electrode at different scanning speeds 1/2 From FIG. 6, it can be seen that the redox peak current is related to v 1/2 A good linear relationship is obtained.
Example 2
The BPNs-SWCNTs/CILE modified electrode obtained in example 1 was used; after airing at room temperature, dripping HRP on the BPNs-SWCNTs/CILE electrode to obtain an HRP/BPNs-SWCNTs/CILE modified electrode; and then dripping Nafion onto the electrode, naturally airing, and preparing the modified electrode Nafion/HRP/BPNs-SWCNTs/CILE. Other modified electrodes Nafion/HRP/CILE, nafion/HRP/SWCNTs/CILE and Nafion/HRP/BPNs/CILE were prepared by the same method. All electrodes were not stored in a refrigerator at 4 c for refrigeration.
Test example 2
Electrochemical performance tests were performed using the Nafion/HRP/BPNs-SWCNTs/CILE, nafion/HRP/SWCNTs/CILE, and Nafion/HRP/BPNs/CILE electrodes prepared in example 2 as working electrodes, saturated calomel electrode as reference electrode, platinum wire as counter electrode, and the three electrodes in 0.1mmol/L PBS using cyclic voltammetry.
FIG. 8 is a cyclic voltammogram of a different modified electrode in 0.1mmol/L PBS in example 2. The absence of redox peaks in the scan potential range on figure CILE (curve a) indicates that no redox reaction has occurred at the electrode surface. Asymmetric redox peaks appear on Nafion/HRP/CILE (curve b) due to slow direct electron transfer of HRP to the electrode. On Nafion/HRP/SWCNTs/CILE (curve c) and Nafion/HRP/BPNs/CILE (curve d), all redox peak currents increase due to the presence of highly conductive SWCNTs and BPNs on the electrode surface improving the conductivity of the electrode interface with increasing electron transfer rate. On Nafion/HRP/BPNs-SWCNTs/CILE (curve e), the redox current signal increased significantly, with a pair of aligned reversible redox peaks at-0.205 and-0.153V, and a peak-to-peak potential (Δep) of 52 mV, indicating that the presence of BPNs-SWCNTs composites with good biocompatibility and high conductivity promoted the rate of electron transfer by HRP. The redox peak current ratio (Ipc/Ipa) was 1.23, indicating that the electrochemical process is quasi-reversible.
FIG. 9 is a graph showing the pH change of Nafion/HRP/BPNs-SWCNTs/CILE in example 2, from which it can be seen that the redox peak potential gradually moves in the negative direction as the pH increases. Indicating that the redox peak current is maximum at pH 4.0.
Test example 3
The Nafion/HRP/BPNs-SWCNTs/CILE electrode prepared in example 2 was used as a working electrode, a saturated calomel electrode was used as a reference electrode, a platinum wire was used as a counter electrode, and the three electrodes were placed in 0.1mmol/L PBS and tested for electrocatalytic performance by cyclic voltammetry.
FIG. 10 is a cyclic voltammogram of electrocatalytic reduction reactions of modified electrodes Nafion/HRP/BPNs-SWCNTs/CILE at different concentrations of TCA of example 2. With the addition of TCA, a new reduction peak appears near-0.485-V, the reduction peak current gradually increases and the oxidation peak current gradually decreases until disappearing, the linear range is 4.0-810.0 mmol/L, and the detection limit is 1.3 mmol/L (3 sigma). HRP has higher catalytic activity in the BPNs-SWCNTs composite material, and the BPNs-SWCNTs has good biocompatibility.
FIG. 11 shows the modified electrode Nafion/HRP/BPNs-SWCNTs/CILE of example 2 at different concentrations of NaNO 2 Cyclic voltammograms of electrocatalytic reduction reactions. With NaNO 2 A new reduction peak appears near-0.460V and follows the NaNO 2 When the addition amount is increased, the reduction peak current is obviously increased, the oxidation peak current is gradually reduced, and when NaNO 2 At a concentration ranging from 0.8 to 49.6 mmol/L, the catalytic reduction peak current and NaNO 2 Is linear in concentration when NaNO 2 When the concentration of (C) exceeds 49.6 mmol/L, the reduction peak current remains substantially unchanged, and the detection limit is 0.27 mmol/L (3σ). Apparent Mi's constant (K) M app ) 8.37 mmol/L, indicating Nafion/HRP/BPNs-SWCNTs/CILE vs. NaNO 2 Has good electrocatalytic performance.
FIG. 12 shows the modified electrode Nafion/HRP/BPNs-SWCNTs/CILE of example 2 at different concentrations of H 2 O 2 Cyclic voltammograms of electrocatalytic reduction reactions. Reduction peak current with H 2 O 2 Increasing the concentration, and reducing the peak current and H when the concentration is in the range of 1.0-40.0 mmol/L 2 O 2 The concentration is in good linear relation, the detection limit is 0.33 mmol/L (3 sigma), K M app At 7.88 mmol/L, indicating Nafion/HRP/BPNs-SWCNTs/CILE vs H 2 O 2 Has better electrocatalytic effect, and the HRP maintains better biological activity in the BPNs-SWCNTs composite material.
Although the foregoing embodiments have been described in some, but not all embodiments of the invention, other embodiments may be obtained according to the present embodiments without departing from the scope of the invention.

Claims (3)

1. The preparation method of the horseradish peroxidase (HRP) electrochemical sensor constructed by the thin-layer Black Phosphazene (BPNs)/single-wall carbon nano tubes (SWCNTs) composite material is characterized by comprising the following steps of:
(1) Preparation of ionic liquid modified carbon paste electrode (CILE)
1.6g of graphite powder and 0.8g of N-hexylpyridine Hexafluorophosphate (HPPF) 6 ) Mixing, grinding uniformly by using a mortar, filling into a glass electrode tube with the diameter of 4mm, compacting, inserting a copper wire as a lead of the electrode, and polishing the surface of the electrode into a mirror surface on polishing paper before use, wherein the prepared electrode is the ionic liquid modified carbon paste electrode (CILE);
(2) Preparation of thin-layer BPNs-SWCNTs nanocomposite
Mixing multi-layer black phosphorus (BPNPs) and N-methyl pyrrolidone (NMP) in an ice bath by adopting a liquid phase stripping method and a physical mixing method, and performing ultrasonic treatment for 12 hours to prepare a thin-layer BPNs dispersion; the volume ratio of BPNs dispersion liquid to SWCNTs is 1:1, mixing, and carrying out ultrasonic treatment for 4 hours in an environment isolated from oxygen to prepare a thin-layer BPNs-SWCNTs nanocomposite;
(3) In a glove box, putting a CILE and a thin-layer BPNs-SWCNTs nano composite material, opening an air outlet valve to close an air inlet valve, vacuumizing by using a vacuum pump, opening the air inlet valve to close the air outlet valve, and introducing nitrogen;
(4) Dropping BPNs-SWCNTs nano composite material on the surface of CILE to obtain BPNs-SWCNTs/CILE modified electrode;
(5) Dripping HRP on the BPNs-SWCNTs/CILE electrode, naturally airing to obtain an HRP/BPNs-SWCNTs/CILE modified electrode;
(6) Dropping Nafion onto HRP/BPNs-SWCNTs/CILE, naturally airing, and preparing the modified electrode Nafion/HRP/BPNs-SWCNTs/CILE with a sandwich structure.
2. The process according to claim 1, wherein the BPNPs used in step (2) are 5.0mL 1.0mg/mL, and the volume ratio of NMP to NMP is 1:1, mixing and carrying out liquid phase ultrasonic stripping; SWCNTs used were 0.5mg/mL, with a volume ratio of BPNs to SWCNTs of 1:1, mixing and ultrasonic treatment; the BPNs-SWCNTs used in step (4) were 10.0. Mu.L, the HRP used in step (5) was 10.0. Mu.L 15.0mg/mL, and the Nafion used in step (6) was 10.0. Mu.L 0.5%.
3. Use of an electrochemical sensor produced by the production method according to claim 1 or 2, characterized by the following steps:
(a) Respectively preparing trichloroacetic acid, sodium nitrite and hydrogen peroxide solution;
(b) The method comprises the steps of performing electrocatalytic performance test by using Nafion/HRP/BPNs-SWCNTs/CILE as a working electrode, a platinum wire as a counter electrode and silver/silver chloride as a reference electrode through cyclic voltammetry, detecting the relation between oxidation peak current values and the concentration of trichloroacetic acid, sodium nitrite and hydrogen peroxide solutions with different concentrations in a phosphate buffer solution, and establishing a standard curve;
(c) And (3) according to the established standard curve, obtaining a detection range, a detection limit and an apparent Mie constant.
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