TWI768639B - Uric acid sensor and the method for manufacturing the same - Google Patents

Abstract

The present disclosure provides a uric acid sensor, including: a flexible substrate; a working electrode disposed on the flexible substrate, and the working electrode includes: a first metal layer disposed on the flexible substrate; a sensing electrode layer disposed on the first metal layer of a first portion of the working electrode; a silver nanowires layer disposed on the sensing electrode layer; and an enzyme composite layer disposed on the silver nanowires layer, wherein the enzyme composite layer includes a composite of silver nanoparticles and uricase; and a reference electrode disposed on the flexible substrate, and the reference electrode includes a second metal layer.

Description

Uric acid sensor and method of making the same

The embodiments of the present invention relate to a uric acid sensor, in particular, to a uric acid sensor including nanosilver wires and nanosilver particles.

The metabolite of purines produced in human metabolism is uric acid, and certain diseases may lead to elevated uric acid to form hyperuricemia, such as diabetes, obesity, high cholesterol, heart disease, and kidney disease, and hyperuricemia may cause cause gout. Therefore, the determination of uric acid in human blood or urine is an important indicator for clinical diagnosis, especially regarding liver and kidney functions. Today, there are many ways to measure the content of uric acid, mainly including colorimetry and electrochemical methods. Among these methods, the method of quantifying the concentration of uric acid by colorimetry is easily affected by interfering substances in the human body, such as vitamin C, bilirubin, and hemolysis, and thus causes inevitable errors. In addition, the disadvantages of traditional analysis methods include long detection time, complicated operation, expensive equipment, and inability to obtain inspection data that can be output in real time. In recent years, many researchers have developed new analytical techniques in view of the shortcomings of traditional analytical methods, and used electrochemical methods to conduct related research.

Many literatures point out that urate oxidase is especially suitable for electrochemical measurement. The enzyme-based uric acid biomedical sensor uses uric acid oxidase (uricase) as the electrode composition. After uric acid reacts with uric acid oxidase, uric acid is oxidized to allantoin, carbon dioxide (CO ₂ ) and peroxide. Hydrogen (H ₂ O ₂ ), and hydrogen peroxide is then hydrolyzed into hydrogen ions (H ⁺ ) and oxygen (O ₂ ), so the difference in uric acid concentration can be sensed by monitoring the change in the concentration of the generated hydrogen ions.

However, although some sensors using electrochemical methods to measure uric acid content have been reported in the literature in recent years, the sensitivity and stability of the existing uric acid sensors still need to be improved. If the sensitivity, stability, anti-interference ability and other properties of the uric acid enzyme sensor can be improved, it will be helpful to detect the concentration of uric acid in the blood, and promote the development of medical diagnosis and daily health care.

An embodiment of the present invention provides a uric acid sensor, including: a flexible substrate; a working electrode, disposed on the flexible substrate, and the working electrode includes: a first metal layer, disposed on the flexible substrate; a sensing electrode The layer is arranged on the first metal layer at the first end of the working electrode; the nano-silver wire layer is arranged on the sensing electrode layer; and the enzyme compound layer is arranged on the nano-silver wire layer, wherein the enzyme is compounded The layer includes a complex of nano-silver particles and urate oxidase; and a reference electrode is disposed on the flexible substrate, and the reference electrode includes a second metal layer.

An embodiment of the present invention further provides a method for manufacturing a uric acid sensor, including: providing a flexible substrate; forming a first working electrode on the flexible substrate a metal layer and a second metal layer of the reference electrode; on the first metal layer of the working electrode at the first end, a sensing electrode layer of the working electrode is formed; on the sensing electrode layer, a nano-silver wire layer of the working electrode is formed ; and an enzyme composite layer that forms a working electrode on the nano-silver wire layer, wherein the enzyme composite layer includes a composite of nano-silver particles and urate oxidase.

110: Flexible substrate

120: Working electrode

121: first metal layer

121a, 131a: first end

121b, 132b: second end

121c, 131c: end

122: Sensing electrode layer

123: Nano silver wire layer

124: Enzyme complex layer

130: Reference electrode

131: Second metal layer

140: Silver Array Electrode Pattern

150: Insulation layer

150a: First opening

150b: Second opening

The embodiments of the present invention will be described in detail below with reference to the accompanying drawings. It should be noted that, in accordance with standard practice in the industry, the various features are not drawn to scale and are illustrative only. In fact, the dimensions of elements may be arbitrarily enlarged or reduced to clearly characterize the embodiments of the invention.

FIGS. 1A to 1E are top views of the uric acid sensor at various stages of the manufacturing process according to some embodiments of the present disclosure.

FIG. 2 is a diagram illustrating a layered structure of a uric acid sensor according to some embodiments of the present disclosure.

FIG. 3 is a graph showing the relationship between the average sensitivity and linearity of the uric acid sensor for uric acid concentration according to some embodiments of the present disclosure.

FIG. 4 is a graph showing the response voltage of a uric acid sensor with time at a specific uric acid concentration, according to some embodiments of the present disclosure.

FIG. 5 is a graph showing the change of the response voltage of the uric acid sensor with time when various chemical substances are added to the solution to be tested for uric acid according to some embodiments of the present disclosure.

The following disclosure provides many different embodiments or examples to illustrate different components of embodiments of the invention. The following will disclose specific examples of the components and their arrangement of the present specification, so as to simplify the description of the present disclosure. Of course, these specific examples are not intended to limit the present disclosure. For example, if the following summary of the present specification describes that the first part is formed on or above the second part, it means that it includes the embodiment in which the first and second parts are formed in direct contact, and also includes further Additional components may be formed between the first and second components described above, the first and second components being embodiments that are not in direct contact. In addition, the various examples in this disclosure may use repeated reference symbols and/or wording. These repeated symbols or words are used for simplicity and clarity, and are not used to limit the relationships between the various embodiments and/or the configurations.

Furthermore, for convenience in describing the relationship of one element or component to another element or component(s) in the drawings, spatially relative terms such as "below", "below", "lower", "above" may be used , "upper" and similar terms. In addition to the orientation shown in the drawings, spatially relative terms also encompass different orientations of the device in use or operation. When the device is turned in a different orientation (eg, rotated 90 degrees or other orientations), the spatially relative adjectives used therein will also be interpreted according to the turned orientation.

Here, the terms "about", "approximately" and "approximately" generally mean within 20%, preferably within 10%, and more preferably within 5% of a given value or range, or within 3% Within %, or within 2%, or within 1%, or within 0.5%. It should be noted that the quantities provided in the specification are approximate quantities, that is to say, “about”, “approximately” and “approximately” can still be implied without the specific description of “about”, “approximately” and “approximately”. probably” meaning.

1A to 1E are according to some embodiments of the present disclosure, depicting uric acid Top view of the sensor at various stages of the process. Referring to FIG. 1A , a flexible substrate 110 is first provided, and a first metal layer 121 serving as a working electrode 120 and a second metal layer 131 serving as a reference electrode 130 are formed on the flexible substrate 110 .

In some embodiments, the material of the flexible substrate 110 may include polyethylene terephthalate (PET), polysulfone (PES), polyethylene naphthalate (polythylene naphtalate) , PEN), polyimide (polyimide, PI), polycarbonate (polycarbonate, PC) or other suitable soft materials. In some embodiments, the first metal layer 121 and the second metal layer 131 may include silver, gold, platinum, palladium or other suitable metal materials. In a specific embodiment, the first metal layer 121 and the second metal layer 131 are part of a silver array electrode pattern (eg, the silver array electrode pattern 140 in FIG. 2 ). In some embodiments, the first metal layer 121 and the second metal layer 131 may be formed using, for example, screen printing, ink-jet printing, or other suitable techniques.

As shown in FIG. 1A , the first metal layer 121 serving as the working electrode 120 may have a first end portion 121 a , a second end portion 121 b and a neck portion 121 c connecting the first end portion 121 a and the second end portion 121 b . The second metal layer 131 serving as the reference electrode 130 may have a first end portion 131a, a second end portion 131b, and a neck portion 131c connecting the first end portion 131a and the second end portion 131b, wherein the working electrode 120 and the reference electrode 130 are solid separated. In some embodiments, the length of the first end 121a and the second end 121b in the Y-axis direction is smaller than the length of the neck 121c in the Y-axis direction, and the length of the first end 121a and the second end 121b in the X-axis direction The width is greater than the width of the neck 121c in the X-axis direction, the first end 131a and the second end portion 131b in the Y-axis direction are smaller than the length of the neck portion 131c in the Y-axis direction, and the width of the first end portion 131a and the second end portion 131b in the X-axis direction is greater than that of the neck portion 131c in the X-axis direction. width. However, the present disclosure is not limited thereto, and those skilled in the art can adjust the first end portions of the first metal layer 121 serving as the working electrode 120 and the second metal layer 131 serving as the reference electrode 130 according to design requirements 121a and 131a, the second ends 121b and 131b, and the necks 121c and 131c have lengths in the Y-axis direction and widths in the X-axis direction.

Although the flexible substrate 110 shown in FIGS. 1A to 1E has six first metal layers 121 serving as working electrodes 120 and two second metal layers 131 serving as reference electrodes 130 , the present disclosure is not based on this As a limitation, those with ordinary knowledge in the field of the present invention can adjust the number of the first metal layer 121 serving as the working electrode 120 and the number of the second metal layer 131 serving as the reference electrode 130 according to design requirements. The array formed by the plurality of working electrodes 120 and the plurality of reference electrodes 130 can reduce the measurement error of the uric acid sensor.

Next, referring to FIG. 1B , a sensing electrode layer 122 of the working electrode 120 is formed on the first end portion 121 a of the first metal layer 121 . In some embodiments, the sensing electrode layer 122 may include a metal oxide layer. In some embodiments, the metal oxide layer of the sensing electrode layer 122 may include nickel oxide (NiO), zinc oxide (ZnO), titanium dioxide (TiO ₂ ), ruthenium dioxide (RuO ₂ ), indium gallium oxide (indium gallium oxide) zinc oxide (IGZO), copper oxide (CuO), indium-tin oxide (ITO) or other suitable metal oxides. In a specific embodiment, nickel oxide may be used as the material of the metal oxide layer of the sensing electrode layer 122 . Nickel oxide is a p-type semiconductor that is widely used as a uric acid sensor due to its high chemical stability, biocompatibility, high electron transfer properties, non-toxicity, high electrical point, and high electrocatalytic properties. test material. In some embodiments, the metal oxide layer of the sensing electrode layer 122 has a thickness of about 100 nm to about 110 nm.

In other embodiments, the sensing electrode layer 122 may further include a third metal layer (not shown) disposed between the first metal layer 121 and the metal oxide layer. In some embodiments, the third metal layer of the sensing electrode layer 122 may include an aluminum thin film, a silver thin film, a platinum thin film, or other suitable metal thin films. The third metal layer can improve the conductivity of the sensing electrode layer 122 .

In some embodiments, deposition processes such as radio frequency sputtering deposition, chemical vapor deposition (CVD), atomic layer deposition (ALD), plasma assisted chemical deposition may be utilized Vapor deposition (plasma enhanced CVD, PECVD), high density plasma chemical vapor deposition (high density CVD, HDCVD), physical vapor deposition (physical vapor deposition, PVD), spin-on coating (spin-on coating) deposition, One or more other suitable processes or a combination of the foregoing form the metal oxide layer and the third metal layer of the sensing electrode layer 122 . In a specific embodiment, the metal oxide layer of the sensing electrode layer 122 with better thickness uniformity may be deposited by radio frequency sputtering. In addition, in some embodiments, the third metal layer of the sensing electrode layer 122 may also be formed by a thermal evaporation process.

Next, referring to FIG. 1C, nano-silver wires (silver) are formed. The nanowires) layer 123 is disposed on the sensing electrode layer 122 and covers a part of the sensing electrode layer 122 . The nano-silver wire layer 123 includes nano-silver wires, and in some embodiments of the present disclosure, the nano-silver wires can be synthesized using various synthesis methods. In a specific embodiment, a solution including silver nanowires (hereinafter referred to as silver nanowires solution) can be prepared using a polyol process, and then the nanowires can be coated by a drop-coating method. The silver wire solution is coated on the sensing electrode layer 122 to form the nano-silver wire layer 123 . In a specific embodiment, for example, about 2 μL of the nano-silver wire solution can be coated on the sensing electrode layer 122 by a drop coating method to form the nano-silver wire layer 123 .

In the present disclosure, by forming the nano-silver wire layer 123 on the metal oxide layer (eg, nickel oxide layer) of the sensing electrode layer 122 , the adsorption area of the enzyme on the working electrode 120 can be increased. The conductivity can reduce the electron transfer resistance and increase the electron transfer rate, thus improving the sensitivity of the uric acid sensor.

Next, referring to FIG. 1D , an insulating layer 150 is formed, which covers the flexible substrate 110 , the first end portion 121 a and the neck portion 121 c of the first metal layer 121 , and the first end portion 131 a and the neck portion of the second metal layer 131 131c. The insulating layer 150 has a first opening 150 a to expose at least a part of the silver nanowire layer 123 on the sensing electrode layer 122 , and a second opening 150 b to expose the first end 131 a of the second metal layer 131 . As shown in FIG. 1D, the insulating layer 150 does not cover the second end portion 121b of the first metal layer 121 and the second end portion 131b of the second metal layer 131, and the insulating layer 150 also does not cover the second end portion 121b and 131b on the left and right sides and the lower side of the flexible substrate 110 . In some embodiments, insulating layer 150 has a thickness of about 20 μm to about 30 μm, eg, 26 μm. However, The thickness of the insulating layer 150 in the present disclosure is not particularly limited, as long as the insulating layer 150 can have an insulating function between electrodes (working electrode, reference electrode, etc.).

In some embodiments, the insulating layer 150 may include epoxy, polyimide, or other suitable insulating resins. In some embodiments, screen printing, inkjet printing or other suitable processes may be used on the flexible substrate 110 , the first end portion 121 a and the neck portion 121 c of the first metal layer 121 , and the second metal layer 131 . An insulating layer 150 is formed on the one end portion 131a and the neck portion 131c. In some embodiments, the size of the first opening 150a is in the range of about 8 mm ² to about 10 mm ² .

Referring to FIG. 1E and FIG. 2 , FIG. 2 is a layered structure diagram of a uric acid sensor according to some embodiments of the present disclosure. After the insulating layer 150 is formed, the enzyme composite layer 124 of the working electrode 120 is then formed on the nano-silver wire layer 123 located in the first opening 150a, wherein the enzyme composite layer 124 includes silver nanoparticles and uric acid oxidation. enzyme complex.

In some embodiments, the process of forming the enzyme composite layer 124 on the nano-silver wire layer 123 includes combining the nano-silver particles with urate oxidase through a first bonding material to form the nano-silver for the enzyme composite layer 124 A complex of particles with urate oxidase. In some embodiments, the first bonding material may include carbodiimide hydrochloric acid, calcium alginate, or other suitable coupling materials. In some embodiments, the nanosilver particles have a size of about 5 nm to about 15 nm, eg, 10 nm. In some embodiments, the activity of the uricase used in the enzyme composite layer 124 ranges from about 15 U/mL to about 30 U/mL. in a specific real In the examples, a compound such as 11-mercaptoundecanoic acid (MUA) was first used to generate carboxyl groups on the surface of the silver nanoparticles, and then a material including carbodiimide hydrochloride (such as 1- Ethyl-3-(3-dimethylaminopropyl) carbodiimide (1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, EDC)) as the first bonding material, on the surface of silver nanoparticles The carboxyl group of urea is activated to produce an unstable oxy-acetylated isourea (O-acylisourea) intermediate, which is capable of reacting with the amine group of urate oxidase. Finally, the silver nanoparticles and the urate oxidase are mixed with each other, so that the silver nanoparticles and the urate oxidase are bound by covalent bonds, and a complex of the silver nanoparticles and the urate oxidase is formed. In some embodiments, the ratio of the first bonding material to the nanosilver particles used is, for example, a molar ratio of 1:1.

In some embodiments, the process of forming the enzyme composite layer 124 on the nano-silver wire layer 123 further includes combining the nano-silver particles with the urate oxidase complex and the nano-silver wire layer 123 through a second bonding material. In some embodiments, the second bonding material may include glutaraldehyde, 3-Glycidoxypropyl-trimethoxysilane (GPTS), 3-aminopropyltrimethoxysilane silane (γ-APTES) or other suitable bonding materials. The above-mentioned second bonding material can generate functional groups, such as hydroxyl groups, on the silver nanowire layer 123, so that the complex of the silver nanoparticle and urate oxidase can be fixed on the silver nanowire layer 123 by covalent bonding. . The second bonding material used here is for bonding the urate oxidase in the enzyme composite layer 124 with the silver nanoparticles and the nanosilver wires in the nanosilver wire layer 123 , and especially with the enzyme composite layer 124 . Covalent bond with the hydroxyl group in the nanosilver wire layer 123 Knot.

In a specific embodiment, before binding the nanosilver particle and urate oxidase complex to the nanosilver wire layer 123, the second binding material (eg, glutaraldehyde) is firstly combined with, eg, 1:4 The volume ratio was dissolved in toluene, and then the solution containing the second bonding material was dropped on the nano-silver wire layer 123 and placed in a refrigerator at 4° C. to dry for 12 hours. The volume of the solution containing the second bonding material was between about 2 μL to about 5 μL. Then, the solution containing the complex of nano-silver particles and urate oxidase is filled on the nano-silver wire layer 123 in the first opening 150a of the insulating layer 150 by the drop coating method, and is placed in a refrigerator at 4° C. to dry for 12 hours To form the enzyme complex layer 124, the volume of the solution containing the complex of the silver nanoparticles and the uricase is between about 2 μL and about 5 μL.

By binding the nano-silver particles to urate oxidase, the nano-silver particles can be used as a carrier of urate oxidase, which is beneficial to the preservation of urate oxidase. In addition, in the uric acid sensor of the present disclosure, the nano-silver particles can increase the specific surface area of the enzyme composite layer 124, so that the interface area for electron transfer becomes larger, thereby increasing the catalytic properties of uric acid oxidase, thereby improving uric acid sensing. Sensitivity of the device.

In addition, according to some embodiments of the present disclosure, by using the drop coating method to form the nano-silver wire layer 123 and the enzyme composite layer 124 in the working electrode 120, the tedious process steps can be avoided in the process of manufacturing the uric acid sensor, Reduce manufacturing time and cost of uric acid sensors.

The uric acid sensor manufactured in the present disclosure performs measurement through the reaction between the uric acid oxidase in the enzyme composite layer 124 and the uric acid in the sample to be measured. The catalytic reaction of urate oxidase generates hydrogen ions, which changes the pH value of the environment around the sensing electrode layer. In turn, the surface potential of the sensing electrode layer is affected. By measuring standard samples with different uric acid concentrations, the regression curve of the relationship between the uric acid concentration and the response voltage can be calculated, and then the concentration of uric acid in the sample to be tested can be obtained.

FIG. 3 is a graph showing the relationship between the average sensitivity and linearity of the uric acid sensor to the uric acid concentration according to some embodiments of the present disclosure, wherein the response voltage is obtained by using the uric acid sensor manufactured in the present disclosure in the presence of Uric acid in phosphate buffered solution. The average sensitivity described herein is defined as the change in response voltage (mV/(mg/dL)) per 1 mg/dL change in uric acid concentration. As shown in FIG. 3, the uric acid sensor manufactured according to the embodiment of the present disclosure can detect the uric acid concentration in the range of 2 mg/dL to 10 mg/dL, wherein the average sensitivity of the uric acid sensor is 49.322 mV/ (mg/dL) and the linearity is 0.980, the measurement linearity of the uric acid sensor is close to 1, indicating that it has accurate sensing performance.

FIG. 4 is a graph illustrating a response voltage of a uric acid sensor with time at a specific uric acid concentration according to some embodiments of the present disclosure, wherein the above-mentioned response voltage is a uric acid sensor fabricated in the present disclosure. Measured in phosphate buffered solution containing uric acid. The results show that after adding 6 mg/dL of uric acid to the phosphate buffer solution, the response voltage of the uric acid sensor manufactured according to the embodiments of the present disclosure rises sharply, showing its fast response time, and the response voltage after the addition of uric acid has The low drift rate of 1.023mV/hr indicates that the uric acid sensor in the present disclosure has a stable response voltage.

FIG. 5 is a graph showing the change of the response voltage of the uric acid sensor with time when various chemical substances are added to the solution to be tested for uric acid according to some embodiments of the present disclosure. , to evaluate the anti-jamming properties of the uric acid sensor fabricated according to the embodiments of the present disclosure, wherein the above-mentioned response voltage is measured in a phosphate buffer solution. As shown in Fig. 5, 0.3 mM uric acid was first added to the phosphate buffer solution in step (1), resulting in a sudden increase in the response voltage. After the response voltage is stable, in steps (2), (3), (4), (5), (6), urea (5 mM), glucose (6 mM), ascorbic acid (0.02 mM), dopamine (0.065 mM) ), and lactic acid (0.8 mM) were added to phosphate buffer solution to verify the anti-interference effect of the uric acid sensor fabricated according to the embodiments of the present disclosure. When urea, glucose, ascorbic acid, dopamine, and lactate were added respectively, the response voltage did not change significantly. However, when 0.6mM uric acid solution was added, the response voltage suddenly increased again, indicating that the uric acid sensor has specificity for uric acid. , and has high sensitivity to the concentration change of uric acid solution.

As described above, the present disclosure provides a uric acid sensor and a method for manufacturing the same. By including materials such as nano-silver wires and nano-silver particles in the working electrode, the uric acid sensor can cover the uric acid concentration range of human blood. (2mg/dL to 10mg/dL) with high sensitivity, and has a fast response time, long-term stability, and good anti-interference ability. In addition, the manufacturing method provided by the present disclosure also includes the use of simple process steps (eg, techniques such as drop coating) to complete the uric acid sensor, which can avoid the use of tedious process steps and reduce the manufacturing time and cost of the uric acid sensor. There is considerable potential for the field of uric acid sensing.

Although the present invention is disclosed in the foregoing several preferred embodiments, it is not intended to limit the present invention. Those with ordinary knowledge in the technical field to which the present invention pertains may make some changes and modifications without departing from the spirit and scope of the present invention. Therefore, the present invention The scope of protection shall be determined by the scope of the appended patent application.

110: Flexible substrate

121: first metal layer

122: Sensing electrode layer

123: Nano silver wire layer

124: Enzyme complex layer

131: Second metal layer

140: Silver Array Electrode Pattern

150: Insulation layer

Claims (15)

A uric acid sensor, comprising: a flexible substrate; a working electrode disposed on the flexible substrate, and the working electrode comprising: a first metal layer disposed on the flexible substrate; a sensor A measuring electrode layer is arranged on the first metal layer located at a first end of the working electrode; a nano-silver wire layer is arranged on the sensing electrode layer; and an enzyme compound layer is arranged on the nanometer on the silver wire layer, wherein the enzyme composite layer includes a composite of nano-silver particles and urate oxidase; and a reference electrode disposed on the flexible substrate, and the reference electrode includes a second metal layer, wherein The working electrode is physically separated from the reference electrode. The uric acid sensor of claim 1, wherein the first metal layer and the second metal layer are a silver array electrode pattern. The uric acid sensor of claim 1, wherein the sensing electrode layer comprises a metal oxide. The uric acid sensor of claim 3, wherein the metal oxide comprises nickel oxide. The uric acid sensor of claim 1, further comprising an insulating layer covering the flexible substrate, the working electrode and the reference electrode, wherein the insulating layer has a first opening to expose the first end of the working electrode part of , and has a second opening A first end of the reference electrode is exposed, wherein the enzyme compound layer is located in the first opening. The uric acid sensor of claim 1, wherein the insulating layer does not cover a second end of the working electrode and a second end of the reference electrode. The uric acid sensor of claim 1, wherein the complex of the nano-silver particles and the urate oxidase in the enzyme complex layer is covalently bonded to the nano-silver particles and the urate oxidase through a first bonding material Formed, the first bonding material includes carbodiimide hydrochloric acid. The uric acid sensor of claim 1, wherein the enzyme composite layer is fixed to the silver nanowire layer through a second bonding material, and the second bonding material includes glutaraldehyde. A method of manufacturing a uric acid sensor, comprising: providing a flexible substrate; forming a first metal layer of a working electrode and a second metal layer of a reference electrode on the flexible substrate; A sensing electrode layer of the working electrode is formed on the first metal layer at a first end; a nano-silver wire layer of the working electrode is formed on the sensing electrode layer; and a nano-silver wire layer is formed on the sensing electrode layer An enzyme composite layer of the working electrode is formed on the wire layer, wherein the enzyme composite layer includes a composite of nano-silver particles and uric acid oxidase. The method for manufacturing a uric acid sensor as claimed in claim 9, wherein the process of forming the sensing electrode layer of the working electrode further comprises using a radio frequency sputtering deposition process to form the sensing electrode layer. The method for manufacturing a uric acid sensor as claimed in claim 9, wherein the first metal layer and the second metal layer are part of a silver array electrode pattern, and the silver array electrode pattern is formed by a screen printing technique ) formed. The method for manufacturing a uric acid sensor as claimed in claim 9, before forming the enzyme composite layer of the working electrode, further comprising forming an insulating layer covering the flexible substrate, the working electrode and the reference electrode, wherein the The insulating layer has a first opening exposing a part of the first end of the working electrode, and has a second opening exposing a first end of the reference electrode. The manufacturing method of the uric acid sensor according to claim 12, wherein the process of forming the insulating layer further comprises forming the insulating layer on the flexible substrate, the working electrode and the reference electrode by using a screen printing technique. The method for manufacturing a uric acid sensor according to claim 9, wherein the process of forming the nanosilver wire layer of the working electrode comprises applying a drop-coating method to the nanosilver wire on the working electrode. on the sensing electrode layer. The method for manufacturing a uric acid sensor as claimed in claim 12, wherein the process of forming the enzyme composite layer of the working electrode comprises filling the composite solution of nano-silver particles and urate oxidase on the insulating layer by a drop coating method. on the nanosilver wire layer in the first opening.

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