CN113533459A - Method for preparing uric acid bioelectrochemical sensor by using zirconium phosphate - Google Patents
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- CN113533459A CN113533459A CN202110798283.9A CN202110798283A CN113533459A CN 113533459 A CN113533459 A CN 113533459A CN 202110798283 A CN202110798283 A CN 202110798283A CN 113533459 A CN113533459 A CN 113533459A
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- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 title claims abstract description 58
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 title claims abstract description 58
- 229940116269 uric acid Drugs 0.000 title claims abstract description 58
- 238000000034 method Methods 0.000 title claims abstract description 24
- 229910000166 zirconium phosphate Inorganic materials 0.000 title claims abstract description 24
- LEHFSLREWWMLPU-UHFFFAOYSA-B zirconium(4+);tetraphosphate Chemical compound [Zr+4].[Zr+4].[Zr+4].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O LEHFSLREWWMLPU-UHFFFAOYSA-B 0.000 title claims abstract description 23
- 239000000243 solution Substances 0.000 claims abstract description 46
- 108010092464 Urate Oxidase Proteins 0.000 claims abstract description 35
- 229920001661 Chitosan Polymers 0.000 claims abstract description 24
- VDZOOKBUILJEDG-UHFFFAOYSA-M tetrabutylammonium hydroxide Chemical compound [OH-].CCCC[N+](CCCC)(CCCC)CCCC VDZOOKBUILJEDG-UHFFFAOYSA-M 0.000 claims abstract description 18
- 239000008055 phosphate buffer solution Substances 0.000 claims abstract description 15
- 239000002131 composite material Substances 0.000 claims abstract description 13
- 238000003756 stirring Methods 0.000 claims abstract description 12
- 238000001035 drying Methods 0.000 claims abstract description 11
- 229910021397 glassy carbon Inorganic materials 0.000 claims abstract description 10
- 238000006243 chemical reaction Methods 0.000 claims abstract description 7
- 238000002156 mixing Methods 0.000 claims abstract description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 10
- 239000011259 mixed solution Substances 0.000 claims description 6
- 229960000583 acetic acid Drugs 0.000 claims description 5
- 239000012362 glacial acetic acid Substances 0.000 claims description 5
- 238000009210 therapy by ultrasound Methods 0.000 claims description 4
- 238000003760 magnetic stirring Methods 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 2
- 239000000843 powder Substances 0.000 abstract description 13
- 238000001514 detection method Methods 0.000 abstract description 8
- 230000035945 sensitivity Effects 0.000 abstract description 6
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 238000002484 cyclic voltammetry Methods 0.000 description 6
- 239000007864 aqueous solution Substances 0.000 description 3
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 3
- 230000029142 excretion Effects 0.000 description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 3
- 235000019796 monopotassium phosphate Nutrition 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 3
- 239000001103 potassium chloride Substances 0.000 description 3
- 235000011164 potassium chloride Nutrition 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000012086 standard solution Substances 0.000 description 3
- 229910021642 ultra pure water Inorganic materials 0.000 description 3
- 239000012498 ultrapure water Substances 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 201000005569 Gout Diseases 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 229940075397 calomel Drugs 0.000 description 2
- ZOMNIUBKTOKEHS-UHFFFAOYSA-L dimercury dichloride Chemical compound Cl[Hg][Hg]Cl ZOMNIUBKTOKEHS-UHFFFAOYSA-L 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 238000002798 spectrophotometry method Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000012876 carrier material Substances 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 150000002391 heterocyclic compounds Chemical class 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 238000001075 voltammogram Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for preparing a uric acid bioelectrochemical sensor by using zirconium phosphate, belonging to the technical field of electrochemical biosensors; dispersing the prepared alpha-zirconium phosphate into tetrabutylammonium hydroxide solution to strip the tetrabutylammonium hydroxide solution, dissolving uricase in phosphate buffer solution with PH =7.4 to obtain uricase solution, mixing the stripped zirconium phosphate and the uricase solution at room temperature, magnetically stirring for reaction, and centrifugally drying after the reaction is finished to obtain alpha-ZrP/UOXA composite material; the prepared alpha-ZrP/UOXAdding the powder into 0.5wt% chitosan solution, transferring the solution to the surface of a treated glassy carbon electrode, and drying the electrode to obtain the zirconium phosphate uricase biosensor. The uric acid bioelectrochemical sensor is used for detecting uric acid, and has the advantages of low detection lower limit, high sensitivity and excellent performance.
Description
Technical Field
The invention relates to a method for preparing a uric acid bioelectrochemical sensor by using zirconium phosphate, belonging to the technical field of bioelectrochemistry.
Background
Uric acid is a heterocyclic compound containing carbon, nitrogen, oxygen and hydrogen, the generation amount and the excretion amount of uric acid in a body are unbalanced, so that diseases caused by rise of uric acid in blood are the main cause of gout, and under a normal condition, the uric acid in the body is about 1200 mg, about 600 mg is newly generated every day, 600 mg is excreted, and the uric acid is in a balanced state. However, if too much uric acid is produced in vivo and excretion is delayed or the uric acid excretion mechanism is degraded, the uric acid in vivo is retained too much, and when the blood uric acid concentration is more than 7 mg/l, the body fluid of a human body turns acidic, the normal function of human body cells is affected, and gout can be caused if the normal function of the cells of the human body is influenced. Uric acid has electrochemical activity, and a uric acid bioelectrochemical sensor can be used for detecting the content of uric acid, and the technical problems of low detection lower limit value, low sensitivity and few types of the uric acid bioelectrochemical sensor are always urgently needed to be solved.
Disclosure of Invention
The invention aims to provide a method for preparing a uric acid bioelectrochemical sensor by using zirconium phosphate, which has simple process and quick and convenient operation, and the obtained composite material can be used for the uricase bioelectrochemical biosensor, has simple preparation and excellent performance, can quickly detect the concentration of uric acid in a solution, and specifically comprises the following steps:
(1) mixing the alpha-ZrP water solution and tetrabutylammonium hydroxide solution, magnetically stirring to completely strip the alpha-ZrP into single lamellar alpha-ZrP, performing ultrasonic treatment, and performing centrifugal separation.
(2) Magnetically stirring the alpha-ZrP stripped in the step (1) and uricase solution at room temperature for reaction, and centrifugally drying after the reaction is finished to obtain alpha-ZrP/UOXA composite material.
(3) alpha-ZrP/UOXAnd (3) dissolving the composite material in a chitosan solution, transferring the mixed solution to the surface of the treated glassy carbon electrode, and drying to obtain the uric acid bioelectrochemical sensor.
Preferably, the molar ratio of the alpha-ZrP to the tetrabutylammonium hydroxide in the step (1) is 1:1 to 1: 1.5.
Preferably, the magnetic stirring time in the step (1) is 10-24 h, and the ultrasonic treatment time is 15-40 min.
Preferably, the uricase solution in the step (2) of the invention is prepared by dissolving uricase in a phosphate buffer solution, wherein the concentration of uricase in the uricase solution is 1-3 mg/mL, and the pH of the phosphate buffer solution is = 7.4.
Preferably, the mass ratio of the alpha-ZrP to the uricase in the step (2) is 1: 1-1: 2.
Preferably, the reaction time of the magnetic stirring in the step (2) is 3-5 h.
Preferably, in the step (3), the mass percentage concentration of the chitosan solution is 0.5-1 wt%, and the solvent of the chitosan solution is glacial acetic acid solution with the mass percentage concentration of 1.0%; alpha-ZrP/UOXThe mass ratio of the composite material to the chitosan is 1: 1-2: 1.
Preferably, the amount of the mixture solution to be removed in the step (3) of the present invention is 5. mu.L, and the drying temperature is 0 to 4 ℃.
The uric acid bioelectrochemical sensor prepared by the method is subjected to electrochemical behavior test: ZrP/UOXAnd measuring a cyclic voltammetry curve of the modified glassy carbon electrode serving as a working electrode, a calomel electrode serving as a reference electrode and a Pt electrode serving as a counter electrode to represent the electrochemical response of the modified electrode in a uric acid solution.
The uricase carrier material zirconium phosphate used by the invention has larger specific surface area, stronger chemical stability and excellent lamellar structure; the stable layered structure of zirconium phosphate is used for fixing biological enzyme, the carrier of uricase is used in a uric acid bioelectrochemical sensor, uric acid solution with unknown concentration is rapidly detected through the conversion of electrochemical signals, and the uric acid sensor with low detection lower limit and high sensitivity is provided.
The invention has the beneficial effects that:
(1) the uric acid bioelectrochemical sensor prepared by the invention can be a rapid and efficient detection method for uric acid solution with unknown solubility, and has low detection lower limit and high sensitivity.
(2) The preparation process is easy to control, the experimental operation process only needs to be carried out by an electrochemical workstation, other large-scale equipment is not needed, and the operation is simple and quick.
Drawings
FIG. 1 shows α -ZrP/UO obtained in example 1 of the present inventionXMethod for preparing uric acid bioelectrochemical sensor from composite material in uric acid solutions with different concentrationsCyclic voltammograms.
FIG. 2 shows α -ZrP/UO obtained in example 1 of the present inventionXThe uric acid bioelectrochemical sensor prepared from the composite material is a linear fitting graph of the maximum current and the concentration of an oxidation peak corresponding to a cyclic voltammetry curve under different uric acid concentrations.
Detailed Description
The invention will be described in more detail with reference to the following figures and examples, but the scope of the invention is not limited thereto.
Example 1
A method for preparing a uric acid bioelectrochemical sensor by using zirconium phosphate specifically comprises the following steps:
(1) adding 0.5mmol of layered zirconium phosphate powder into 10.0mL of ultrapure water, adding 0.5mmol of 40wt% tetrabutylammonium hydroxide aqueous solution (TBAOH), magnetically stirring for 15h, and ultrasonically treating for 20min to completely strip the layered zirconium phosphate.
(2) Phosphate Buffer Solution (PBS) having pH =7.4 was prepared using 0.24g of potassium dihydrogenphosphate, 1.44g of disodium hydrogenphosphate, 8.0g of sodium chloride, and 0.2g of potassium chloride, and 6.3mg of uricase was dissolved in 3mL of phosphate buffer solution to prepare a uricase solution.
(3) Mixing all the uricase solution in the step (2) with 6.3mg of the peeled zirconium phosphate in the step (1), magnetically stirring for 3 hours at room temperature, and centrifuging for 3 times to separate the free uricase from the immobilized uricase.
(4) Weighing chitosan powder, adding the chitosan powder into 1.0% glacial acetic acid solution, and preparing 0.5wt% chitosan solution; then alpha-ZrP/UO is addedXThe powder is dissolved in the chitosan solution, wherein alpha-ZrP/UOXThe mass ratio of the composite material to the chitosan is 2: 1.
(5) Pretreating the glassy carbon electrode, transferring 5 mu L of (4) mixed solution on the surface of the glassy carbon electrode, and drying at 4 ℃ to obtain the alpha-ZrP/UOXThe modified electrode is the uric acid bioelectrochemical sensor.
The alpha-ZrP/UO described in this exampleXThe performance test of the modified electrode specifically comprises the following steps:
will be in step (5) of example 1The obtained alpha-ZrP/UOXThe modified electrode, the calomel reference electrode and the Pt counter electrode form a three-electrode system, and cyclic voltammetry tests are carried out in PBS (phosphate buffer) containing uric acid (5, 10,50,100,200,300 and 400 mu mol/L) with different concentrations, so that cyclic voltammetry curves can be obtained, and are shown in figure 1. Linearly fitting the maximum current and the concentration of the corresponding oxidation peak under different uric acid concentrations to obtain a standard curve, as shown in FIG. 2; the zirconium phosphate uricase sensor has good linear relation in the range of 0.1 mu mol/L to 400 mu mol/L and the correlation coefficient R2=0.982, with a detection limit of 0.03 μmol/L (S/N = 3), indicating that the uric acid bioelectrochemical sensor has a low detection limit and high sensitivity.
The uric acid solution with unknown concentration can be quickly known only by knowing the maximum current corresponding to the oxidation peak in the cyclic voltammetry curve under the same condition and corresponding to the standard curve.
The alpha-ZrP/UO described in this exampleXVerification experiment of accuracy of modified electrode:
(1) preparing a uric acid standard solution with the concentration of 250 mu mol/L.
(2) alpha-ZrP/UO prepared in example 1XAnd (3) performing cyclic voltammetry on the uric acid standard solution in the modified electrode pair (1), and obtaining the concentration measured by using an electrochemical biosensor method by contrasting with a standard curve graph 2.
(3) The uric acid standard solution was measured by ultraviolet spectrophotometry, and the concentration measured in (2) was compared, as shown in table 1.
TABLE 1 comparison of the bioelectrochemical sensor method with the UV spectrophotometry method
As can be seen from Table 1, the relative error of the uric acid bioelectrochemical sensor prepared by the method is smaller than that of an ultraviolet spectrophotometry.
Example 2
A method for preparing a uric acid bioelectrochemical sensor by using zirconium phosphate specifically comprises the following steps:
(1) adding 0.5mmol of layered zirconium phosphate powder into 10.0mL of ultrapure water, adding 0.75mmol of 40wt% tetrabutylammonium hydroxide aqueous solution (TBAOH), magnetically stirring for 24h, and ultrasonically treating for 15min to completely strip the layered zirconium phosphate.
(2) Phosphate Buffer Solution (PBS) having pH =7.4 was prepared using 0.24g of potassium dihydrogenphosphate, 1.44g of disodium hydrogenphosphate, 8.0g of sodium chloride, and 0.2g of potassium chloride, and 6.3mg of uricase was dissolved in 2.1mL of phosphate buffer solution to prepare a uricase solution.
(3) Mixing all the uricase solution in the step (2) with 3.15mg of the peeled zirconium phosphate in the step (1), magnetically stirring for 4 hours at room temperature, and centrifuging for 3 times to separate the free uricase from the immobilized uricase.
(4) Weighing chitosan powder, adding the chitosan powder into 1.0% glacial acetic acid solution, and preparing 0.5wt% chitosan solution; then alpha-ZrP/UO is addedXThe powder is dissolved in the chitosan solution, wherein alpha-ZrP/UOXThe mass ratio of the composite material to the chitosan is 1:1.
(5) Pretreating the glassy carbon electrode, transferring 5 mu L of (4) mixed solution on the surface of the glassy carbon electrode, and drying at 4 ℃ to obtain the alpha-ZrP/UOXThe modified electrode is the uric acid bioelectrochemical sensor.
Example 3
A method for preparing a uric acid bioelectrochemical sensor by using zirconium phosphate specifically comprises the following steps:
(1) adding 0.5mmol of layered zirconium phosphate powder into 10.0mL of ultrapure water, adding 0.6mmol of 40wt% tetrabutylammonium hydroxide aqueous solution (TBAOH), magnetically stirring for 10h, and ultrasonically treating for 40min to completely strip the layered zirconium phosphate.
(2) Phosphate Buffer Solution (PBS) having pH =7.4 was prepared using 0.24g of potassium dihydrogenphosphate, 1.44g of disodium hydrogenphosphate, 8.0g of sodium chloride, and 0.2g of potassium chloride, and 6.3mg of uricase was dissolved in 6.3mL of phosphate buffer solution to prepare a uricase solution.
(3) Mixing all the uricase solution in the step (2) with 4.2mg of the peeled zirconium phosphate in the step (1), magnetically stirring for 5 hours at room temperature, and centrifuging for 3 times to separate the free uricase from the immobilized uricase.
(4) Weighing chitosan powder, adding the chitosan powder into 1.0% glacial acetic acid solution, and preparing 0.5wt% chitosan solution; then alpha-ZrP/UO is addedXThe powder is dissolved in the chitosan solution, wherein alpha-ZrP/UOXThe mass ratio of the composite material to the chitosan is 1.5: 1.
(5) Pretreating the glassy carbon electrode, transferring 5 mu L of (4) mixed solution on the surface of the glassy carbon electrode, and drying at 4 ℃ to obtain the alpha-ZrP/UOXThe modified electrode is the uric acid bioelectrochemical sensor.
The uric acid bioelectrochemical sensor prepared in the embodiments 2 and 3 is used for detecting uric acid, and has low detection lower limit, high sensitivity and performance similar to that of the embodiment 1.
Claims (8)
1. A method for preparing a uric acid bioelectrochemical sensor by using zirconium phosphate is characterized by comprising the following steps:
(1) mixing the alpha-ZrP water solution and tetrabutylammonium hydroxide solution, magnetically stirring to completely strip the alpha-ZrP into single lamellar alpha-ZrP, and performing ultrasonic treatment and centrifugal separation;
(2) magnetically stirring the alpha-ZrP stripped in the step (1) and uricase solution at room temperature for reaction, and centrifugally drying after the reaction is finished to obtain alpha-ZrP/UOXA composite material;
(3) alpha-ZrP/UOXAnd (3) dissolving the composite material in a chitosan solution, transferring the mixed solution to the surface of the treated glassy carbon electrode, and drying to obtain the uric acid bioelectrochemical sensor.
2. The method for preparing uric acid bioelectrochemical sensor according to claim 1, characterized in that: in the step (1), the molar ratio of alpha-ZrP to tetrabutylammonium hydroxide is 1: 1-1: 1.5.
3. The method for preparing uric acid bioelectrochemical sensor according to claim 1, characterized in that: the magnetic stirring time in the step (1) is 10-24 h, and the ultrasonic treatment time is 15-40 min.
4. The method for preparing uric acid bioelectrochemical sensor according to claim 1, characterized in that: the preparation method of the uricase solution comprises the steps of dissolving uricase in a phosphate buffer solution, wherein the concentration of the uricase in the uricase solution is 1-3 mg/mL, and the pH of the phosphate buffer solution is = 7.4.
5. The method for preparing uric acid bioelectrochemical sensor according to claim 4, characterized in that: the mass ratio of the alpha-ZrP to the uricase in the step (2) is 1: 1-1: 2.
6. The method for preparing uric acid bioelectrochemical sensor according to claim 1, characterized in that: and (3) magnetically stirring for 3-5 h.
7. The method for preparing uric acid bioelectrochemical sensor according to claim 1, characterized in that: in the step (3), the mass percent concentration of the chitosan solution is 0.5-1 wt%, and the solvent of the chitosan solution is glacial acetic acid solution with the mass percent concentration of 1.0%; alpha-ZrP/UOXThe mass ratio of the composite material to the chitosan is 1: 1-2: 1.
8. The method for preparing uric acid bioelectrochemical sensor according to claim 1, characterized in that: the transfer amount of the mixed solution in the step (3) is 5 mu L, and the drying temperature is 0-4 ℃.
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