Wire reference electrode for biosensor for healthcare, method for manufacturing same, and biosensor for healthcare including same

Abstract

Provided is a wire reference electrode for a biosensor for healthcare. The wire reference electrode for a biosensor for healthcare may comprise: a metal wire; a chlorinated layer disposed on the surface of the metal wire, the chlorinated layer being formed by chlorinating the metal which makes up the metal wire; a first protective layer disposed on the surface of the chlorinated layer and physically blocking the movement of the chloride ions (Cl-) from the chlorinated layer; and a second protective layer disposed on the surface of the first protective layer and electrostatically blocking the movement of the chloride ions (Cl-).

Description

Wire reference electrode for a biosensor for healthcare, method of manufacturing the same, and biosensor for healthcare including the same

[0001] The present invention relates to a wire reference electrode for a biosensor for healthcare, a method for manufacturing the same, and a biosensor for healthcare including the same. More specifically, the invention relates to a wire reference electrode for a biosensor for healthcare having excellent stability and repeatability, a method for manufacturing the same, and a biosensor for healthcare including the same.

[0002] In electrochemical measurements, the reference electrode is a key element that enables accurate measurements by providing a stable and constant potential.

[0003] Conventional reference electrodes mainly include a silver / silver chloride (Ag / AgCl) electrode formed inside a glass tube and a potassium chloride (KCl) solution that stabilizes it.

[0004] However, conventional reference electrodes use glass tubes, which are prone to breakage and pose a risk of potassium chloride solution leakage; if the potassium chloride solution leaks when inserted into the human body, it can cause extremely fatal problems.

[0005] Furthermore, conventional reference electrodes are bulky and difficult to handle in small devices or specialized applications, which limits their use.

[0006] Currently, as the application fields of electrochemical equipment expand to include wearable sensors, micro-needle-based electrodes, point-of-care diagnostic devices, and environmental monitoring systems, there is a growing demand for miniaturization of reference electrodes.

[0007] As part of this effort, wire-shaped reference electrodes have recently been actively researched.

[0008] Nafion coating, a fundamental technology applied to wire reference electrodes, has not yet achieved commercial-level performance and stability due to a lack of selective ion permeability, pH sensitivity, potential drift issues caused by changes in electrolyte concentration, and environmental adaptability under various electrolyte conditions.

[0009] For this reason, Nafion coating technology alone is insufficient to fully meet the requirements of various application fields.

[0010] Accordingly, approaches such as electrode surface treatment using nanotechnology, application of polymer electrolyte membranes, and electrolyte management using microfluidic technology are being studied, but some face difficulties in mass production due to complex manufacturing processes and high costs. Therefore, there is a need to develop more innovative miniaturization technologies.

[0011] The technical problem that the present invention aims to solve is to provide a wire reference electrode for a biosensor for healthcare having excellent stability and repeatability, a method for manufacturing the same, and a biosensor for healthcare including the same.

[0012] The technical problems that the present invention aims to solve are not limited to those described above.

[0013] To solve the above-mentioned technical problem, the present invention provides a wire reference electrode for a biosensor for healthcare.

[0014] According to one embodiment, the wire reference electrode for a biosensor for healthcare comprises: a metal wire; a chlorination layer provided on the surface of the metal wire and formed through chlorination of the metal constituting the metal wire; and chloride ions (Cl) provided on the surface of the chlorination layer and formed from the chlorination layer. - A first defense layer that physically blocks the movement of ); and a first defense layer provided on the surface of the first defense layer, wherein the chloride ions (Cl -It may include a second defense layer that electrostatically blocks the movement of ).

[0015] According to one embodiment, the metal wire is made of silver (Ag), and the chlorination layer may be made of silver chloride (AgCl).

[0016] According to one embodiment, the first protective layer may be made of polyvinyl butyral (PVB), and the second protective layer may be made of Nafion.

[0017] According to one embodiment, the wire reference electrode for the biosensor can be an all-solid-state electrode.

[0018] Meanwhile, the present invention provides a biosensor for healthcare.

[0019] According to one embodiment, the healthcare biosensor comprises: a wire reference electrode; and a sensor portion provided on the wire reference electrode and made of a material that responds to a target biomaterial, wherein the wire reference electrode comprises: a metal wire; a chlorination layer provided on the surface of the metal wire and formed through chlorination of the metal constituting the metal wire; and a chloride ion (Cl) provided on the surface of the chlorination layer and formed from the chlorination layer - A first defense layer that physically blocks the movement of ); and a first defense layer provided on the surface of the first defense layer, wherein the chloride ions (Cl - It may include a second defense layer that electrostatically blocks the movement of ).

[0020] In addition, the present invention provides a method for manufacturing a wire reference electrode for a biosensor for healthcare.

[0021] According to one embodiment, the method for manufacturing a wire reference electrode for a biosensor for healthcare comprises: a preparation step of preparing a metal wire; a chlorination layer formation step of forming a chlorination layer on the surface of the metal wire through chlorination of the prepared metal wire; and chloride ions (Cl) from the chlorination layer formed on the surface of the metal wire. - A first defense layer forming step of forming a first defense layer on the surface of the chlorination layer to physically block the movement of ); and the chloride ions (Cl - It may include a step of forming a second defense layer that electrostatically blocks the movement of ) on the surface of the first defense layer.

[0022] According to one embodiment, in the preparation step, a metal wire made of silver (Ag) is prepared, and in the chlorination layer formation step, a chlorination layer made of silver chloride (AgCl) may be formed on the surface of the metal wire.

[0023] According to one embodiment, in the first protective layer formation step, the first protective layer can be formed with polyvinyl butyral (PVB).

[0024] According to one embodiment, the first protective layer formation step may include: a process of preparing a polyvinyl butyral (PVB) solution; a process of preparing a polyvinyl butyral cocktail by dissolving a mixed powder of sodium chloride (NaCl) and silver nitrate (AgNO3) in the polyvinyl butyral (PVB) solution; a process of coating a metal wire having a chlorinated layer formed on its surface with the prepared polyvinyl butyral cocktail; and a process of drying and curing the metal wire coated with the polyvinyl butyral cocktail on its outermost surface so that the first protective layer made of polyvinyl butyral (PVB) is formed on the surface of the chlorinated layer.

[0025] According to one embodiment, in the drying and curing process, the metal wire coated with the polyvinyl butyral cocktail on the outermost surface may be dried for up to 1 hour, and then exposed to a UV lamp for up to 10 minutes.

[0026] According to one embodiment, in the drying and curing process, sodium ions (Na₂C₃) generated when sodium chloride (NaCl) and silver nitrate (AgNO₃) dissociate upon dissolving in the polyvinyl butyral (PVB) solution are + ), chloride ions (Cl - ), silver ions (Ag + ) and nitrate ions (NO3 - At least one of the ions can form a cross-link through chemical interaction with the hydroxyl group (-OH) of the polyvinyl butyral (PVB) polymer chain.

[0027] According to one embodiment, in the step of forming the second defense layer, the second defense layer can be formed with Nafion.

[0028] According to an embodiment of the present invention, a metal wire; a chlorination layer provided on the surface of the metal wire and formed through chlorination of the metal constituting the metal wire; and chloride ions (Cl) provided on the surface of the chlorination layer and formed from the chlorination layer - A first defense layer that physically blocks the movement of ); and a first defense layer provided on the surface of the first defense layer, wherein the chloride ions (Cl - It may include a second defense layer that electrostatically blocks the movement of ).

[0029] Accordingly, a wire reference electrode for a biosensor for healthcare having excellent stability and repeatability, a method for manufacturing the same, and a biosensor for healthcare including the same can be provided.

[0030] That is, according to an embodiment of the present invention, a chlorinated layer made of silver chloride (AgCl) formed by chlorination treatment can provide a stable reference potential, and a first protective layer made of polyvinyl butyral (PVB) can effectively prevent leakage of internal electrolyte while enabling the maintenance of ion conductivity.

[0031] At this time, due to the physical properties of polyvinyl butyral (PVB), excellent durability can be ensured even under repeated mechanical deformation, and stable performance can be maintained even during long-term use.

[0032] In addition, according to an embodiment of the present invention, long-term usability or rapid response can be optimized by adjusting the thickness of the first protective layer made of polyvinyl butyral (PVB) according to the application of the biosensor.

[0033] In addition, according to an embodiment of the present invention, the chemical stability of the Nafion forming the second defense layer provided at the outermost layer is excellent, so reliable results can be obtained even in various measurement environments.

[0034] Meanwhile, according to an embodiment of the present invention, excellent flexibility can be secured due to the wire-shaped structural characteristics, and through this, it can be applied to various application fields such as wearable devices or bio-attached sensors.

[0035] In addition, according to an embodiment of the present invention, a method for manufacturing a wire reference electrode for a biosensor for healthcare can be provided, which has a simple manufacturing process, high reproducibility, enables mass production, and can significantly improve economic efficiency.

[0036] Thus, according to an embodiment of the present invention, a wire reference electrode for a healthcare biosensor, a method for manufacturing the same, and a healthcare biosensor including the same can be provided, which can be utilized in various fields such as body fluid analysis for medical diagnosis and electrolyte concentration measurement.

[0037] In particular, the wire reference electrode for a biosensor for healthcare according to an embodiment of the present invention can be utilized as a platform for various next-generation sensors requiring miniaturization, flexibility, and stability.

[0038] FIG. 1 is a perspective view showing a wire reference electrode for a biosensor for healthcare according to one embodiment of the present invention.

[0039] Figure 2 is a partial cutaway perspective view of Figure 1.

[0040] FIG. 3 is a cross-sectional view showing a wire reference electrode for a biosensor for healthcare according to one embodiment of the present invention.

[0041] FIG. 4 is a flowchart illustrating a method for manufacturing a wire reference electrode for a biosensor for healthcare according to one embodiment of the present invention.

[0042] Figures 5 and 6 are schematic diagrams for explaining step S110 of Figure 4.

[0043] Figures 7 and 8 are schematic diagrams for explaining step S120 of Figure 4.

[0044] Figures 9 and 10 are schematic diagrams for explaining step S130 of Figure 4.

[0045] FIGS. 11 to 13 are schematic diagrams for explaining step S140 of FIG. 4.

[0046] Figure 14 shows the open-circuit potential measurement results for the electrodes produced at each step of manufacturing the wire reference electrode according to Example 1.

[0047] Figure 15 shows the open-circuit potential measurement results for methanol and acetone-based polyvinyl butyral coated electrodes.

[0048] Figure 16 shows the open-circuit potential measurement results for an undried electrode that underwent UV curing immediately without drying after polyvinyl butyral coating.

[0049] Figure 17 shows the results of open-circuit potential measurements for a UV photocurable electrode for 30 minutes after drying the coated polyvinyl butyral.

[0050] Figure 18 shows the results of measuring the open-circuit potential of coated polyvinyl butyral on a UV photocuring electrode for 10 minutes.

[0051] FIGS. 19 and 20 are scanning electron microscope images of a cross-section of a wire reference electrode manufactured according to Example 1.

[0052] FIGS. 21 to 25 are the results of analyzing a wire reference electrode manufactured according to Example 1 using energy dispersive spectroscopy (EDS).

[0053] FIGS. 26 and 27 are results of measuring the concentration of glucose using the time-amplification (CA) method with a glucose sensor fabricated using a wire reference electrode manufactured according to Example 1.

[0054] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the attached drawings. However, the technical concept of the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the embodiments introduced herein are provided to ensure that the disclosed content is thorough and complete and to ensure that the concept of the present invention is sufficiently conveyed to those skilled in the art.

[0055] In this specification, when a component is described as being on another component, it means that it may be formed directly on the other component or that a third component may be interposed between them. Additionally, in the drawings, shapes and sizes are exaggerated for the effective description of the technical content.

[0056] Additionally, although terms such as first, second, third, etc., have been used to describe various components in the various embodiments of this specification, these components should not be limited by such terms. These terms are used merely to distinguish one component from another. Accordingly, what is referred to as the first component in one embodiment may be referred to as the second component in another embodiment. Each embodiment described and illustrated herein also includes its complementary embodiment. Furthermore, in this specification, "and / or" is used to mean including at least one of the components listed before and after it.

[0057] In the specification, singular expressions include plural expressions unless the context clearly indicates otherwise. Furthermore, terms such as "include" or "have" are intended to specify the existence of the features, numbers, steps, components, or combinations thereof described in the specification, and should not be understood as excluding the existence or addition of one or more other features, numbers, steps, components, or combinations thereof. Additionally, in this specification, "connection" is used to include both indirectly connecting multiple components and directly connecting them.

[0058] Additionally, terms such as “…part,” “…unit,” and “module” described in the specification refer to a unit that processes at least one function or operation, and this may be implemented in hardware, software, or a combination of hardware and software.

[0059] Furthermore, in describing the present invention below, if it is determined that a detailed description of related known functions or configurations could unnecessarily obscure the essence of the invention, such detailed description will be omitted.

[0060]

[0061] FIG. 1 is a perspective view showing a wire reference electrode for a biosensor for healthcare according to one embodiment of the present invention, FIG. 2 is a partially cutaway perspective view of FIG. 1, and FIG. 3 is a cross-sectional view showing a wire reference electrode for a biosensor for healthcare according to one embodiment of the present invention.

[0062]

[0063] As illustrated in FIGS. 1 to 3, a wire reference electrode (100) for a biosensor according to one embodiment of the present invention can have excellent stability and repeatability even in extreme environments, and thereby can be provided as a platform for various biosensors that sense biomaterials such as glucose and cholesterol.

[0064] In addition, the wire reference electrode (100) for a biosensor according to one embodiment of the present invention can secure excellent flexibility due to its wire-shaped structural characteristics, and thereby can be applied to various application fields such as wearable devices or bio-attached sensors.

[0065] As such, the wire reference electrode (100) for a biosensor according to one embodiment of the present invention can be utilized in various fields such as body fluid analysis for medical diagnosis and electrolyte concentration.

[0066]

[0067] A wire reference electrode (100) for a biosensor according to one embodiment of the present invention may include a metal wire (110), a chlorinated layer (120), a first protective layer (130), and a second protective layer (140).

[0068]

[0069] The metal wire (110) provides excellent flexibility to the wire reference electrode (100) for the biosensor, so that the wire reference electrode (100) for the biosensor can be utilized in wearable devices or bio-attached sensors.

[0070] According to one embodiment of the present invention, such metal wire (110) may be made of silver (Ag) wire. Silver (Ag) wire has high conductivity and a high surface area, so signal transmission is very fast and efficient, and it also has chemically stable characteristics.

[0071]

[0072] The chlorination layer (120) may be provided on the surface of the metal wire (110). According to one embodiment of the present invention, such a chlorination layer (120) may be formed on the surface of the metal wire (110) through chlorination of the metal forming the metal wire (110). That is, the chlorination layer (120) may be coated on the surface of the metal wire (110) through chlorination of the metal forming the metal wire (110).

[0073] According to one embodiment of the present invention, the chlorination layer (120) may be made of silver chloride (AgCl) through chlorination of silver (Ag) as the metal wire (110) is made of silver (Ag) wire.

[0074] In this way, the chlorinated layer (120), that is, the silver chloride (AgCl) layer formed on the surface of the metal wire (110) through chlorination treatment of the silver (Ag) wire forming the metal wire (110), can provide a stable reference potential.

[0075] Here, this chlorinated layer (120) can be coated with a silver chloride (AgCl) layer on the surface of a silver (Ag) metal wire (110) by chlorination by galvanostatic method in a mixed solution of hydrochloric acid (HCl) and potassium chloride (KCl), with the silver (Ag) metal wire (110) as the working electrode (WE), a platinum (Pt) electrode as the counter electrode (CE), and a commercial silver chloride electrode (Ag / AgCl electrode) as the reference electrode (RE) (see FIG. 7).

[0076]

[0077] The first protective layer (130) can be provided on the surface of the chlorinated layer (120) which is made of silver chloride (AgCl).

[0078] According to one embodiment of the present invention, the first protective layer (130) is formed from chloride ions (Cl₂) from the chlorination layer (120) composed of silver chloride (AgCl). - It can serve as a barrier that physically blocks the movement of ).

[0079] This first protective layer (130) may be made of a polymer compound. According to one embodiment of the present invention, the first protective layer (130) may be made of polyvinyl butyral resin (PVB) synthesized from polyvinyl alcohol (PVA) and butyraldehyde.

[0080] In this way, the first protective layer (130) made of polyvinyl butyral (PVB) is a hydrophobic polymer that forms a dense and rigid network, thereby ionizing chloride ions (Cl -It can be provided as a barrier that can physically block the movement of ).

[0081] Through this, the first protective layer (130) made of polyvinyl butyral (PVB) can effectively prevent leakage of internal electrolyte while maintaining ion conductivity.

[0082] At this time, the first protective layer (130) can ensure excellent durability even under repeated mechanical deformation due to the physical properties of polyvinyl butyral (PVB), and accordingly, stable performance can be maintained even during long-term use.

[0083] Additionally, the thickness of the first protective layer (130) made of polyvinyl butyral (PVB) can be adjusted according to the application of the biosensor to which the wire reference electrode (100) according to one embodiment of the present invention is applied.

[0084] Through this, the long-term usability or rapid response of the biosensor can be optimized.

[0085]

[0086] The second protective layer (140) can be provided on the surface of the first protective layer (130) made of polyvinyl butyral (PVB).

[0087] This second defense layer (140) can form the outermost layer of the wire reference electrode (100) for a biosensor according to one embodiment of the present invention.

[0088] According to one embodiment of the present invention, the second defense layer (140), like the first defense layer (130), is formed from chloride ions (Cl₂) from the chlorination layer (120) made of silver chloride (AgCl). - It can act as a barrier to block the movement of ).

[0089] At this time, the second protective layer (140) is formed from chloride ions (Cl₂) from the chlorination layer (120) composed of silver chloride (AgCl). - The movement of ) can be electrostatically blocked.

[0090] This second protective layer (140) can be made of a polymer compound that can maintain stability even in high temperature and high humidity environments.

[0091] According to one embodiment of the present invention, the second protective layer (140) may be made of Nafion, which is mainly used for coating electrochemical devices or polymer electrolytes, as it maintains stability even in high temperature and high humidity environments.

[0092] The Nafion forming the second defense layer (140) above is a negatively charged sulfonic acid group with chloride ions (Cl - It can electrostatically repel ). In this case, the hydrophobic polytetrafluoroethylene (PTFE) backbone of Nafion can form a physical barrier.

[0093] Accordingly, the wire reference electrode (100) for a biosensor according to one embodiment of the present invention can maintain chemical stability even in extreme environments, and thereby, reliable results can be obtained in various measurement environments.

[0094] Polyvinyl butyral (PVB) forming the first defense layer (130) and Nafion forming the second defense layer (140) contain chloride ions (Cl - It can block external leakage of ).

[0095] Accordingly, the wire reference electrode (100) for a biosensor according to one embodiment of the present invention can be provided as an all-solid electrode that does not require a stabilization solution, unlike conventional reference electrodes that require a stabilization solution.

[0096] Polyvinyl butyral (PVB) forming the first defense layer (130) and Nafion forming the second defense layer (140) can form a double barrier on the outside of the chlorinated layer (120) made of silver chloride (AgCl).

[0097] Accordingly, the physical barrier of polyvinyl butyral (PVB) forming the first defense layer (130) and the electrostatic barrier of Nafion forming the second defense layer (140) can exhibit a synergistic effect, and through this, chloride ions (Cl₂) from the chlorination layer (120) made of silver chloride (AgCl) - It can effectively block the movement of ).

[0098] As a result, the durability and performance of the wire reference electrode (100) for a biosensor according to one embodiment of the present invention can be improved. This means that the wire reference electrode (100) for a biosensor according to one embodiment of the present invention can be utilized as a platform for various biosensors, and it is possible to obtain reliable medical diagnostic results through the applied biosensor.

[0099]

[0100] As described above, a wire reference electrode (100) for a biosensor according to one embodiment of the present invention is provided with a first defense layer (130) made of polyvinyl butyral (PVB) and a second defense layer (140) made of Nafion, which form a double barrier on the outside of a chlorinated layer (120) made of silver chloride (AgCl) coated on the surface of a metal wire (110) made of silver (Ag), so that it can operate stably even in extreme environments.

[0101] That is, according to one embodiment of the present invention, a wire reference electrode (100) for a biosensor for healthcare having excellent stability and repeatability can be provided.

[0102] Here, silver chloride (AgCl) forming a chlorinated layer (120) coated on the surface of a metal wire (110) made of silver (Ag) through chlorination treatment can provide a stable reference potential.

[0103] In addition, the polyvinyl butyral (PVB) forming the first defense layer (130) can maintain the internal electrolyte of the wire reference electrode (100) for a biosensor according to one embodiment of the present invention at a constant level, thereby improving the repeatability of the biosensor to which the wire reference electrode (100) for a biosensor is applied.

[0104] And due to the excellent chemical stability of Nafion forming the second defense layer (140) above, the wire reference electrode (100) for a biosensor according to one embodiment of the present invention can maintain chemical stability even in extreme environments.

[0105] At this time, the wire reference electrode (100) for a biosensor according to one embodiment of the present invention can secure excellent flexibility due to its wire-shaped structural characteristics, and through this, can be applied to various application fields such as wearable devices or bio-attached sensors.

[0106] In this way, according to one embodiment of the present invention, a wire reference electrode (100) for a biosensor for healthcare, which can be utilized in various fields such as body fluid analysis for medical diagnosis and electrolyte concentration measurement, may be provided.

[0107] In particular, the wire reference electrode (100) for a biosensor for healthcare according to an embodiment of the present invention can be utilized as a platform for various next-generation sensors that require miniaturization, flexibility, and stability.

[0108]

[0109] In this way, a wire reference electrode (100) for a biosensor for healthcare, comprising a first defense layer (130) made of polyvinyl butyral (PVB) and a second defense layer (140) made of Nafion, which form a double barrier on the outside of a chlorinated layer (120) made of silver chloride (AgCl) coated on the surface of a metal wire (110) made of silver (Ag), can be applied to a biosensor for healthcare.

[0110] That is, the above-mentioned healthcare biosensor may include the wire reference electrode (100) and the sensor part.

[0111] Although not illustrated, the sensor portion may be provided on a wire reference electrode (100) according to one embodiment of the present invention. This sensor portion may be made of a material that responds to target biomaterials such as glucose, cholesterol, and cortisol, for example.

[0112] Various sensor materials may be formed on a wire reference electrode (100) according to one embodiment of the present invention to form a biosensor for healthcare.

[0113] In this way, the wire reference electrode (100) for a biosensor according to one embodiment of the present invention can be provided as a platform capable of forming sensor materials for various electrochemical biosensors.

[0114]

[0115] Hereinafter, a method for manufacturing a wire reference electrode for a biosensor for healthcare according to one embodiment of the present invention will be described with reference to FIGS. 4 to 13.

[0116]

[0117] FIG. 4 is a flowchart illustrating a method for manufacturing a wire reference electrode for a biosensor for healthcare according to an embodiment of the present invention, FIG. 5 and FIG. 6 are schematic diagrams for explaining step S110 of FIG. 4, FIG. 7 and FIG. 8 are schematic diagrams for explaining step S120 of FIG. 4, FIG. 9 and FIG. 10 are schematic diagrams for explaining step S130 of FIG. 4, and FIG. 11 to FIG. 13 are schematic diagrams for explaining step S140 of FIG. 4.

[0118]

[0119] Referring to FIG. 4, a method for manufacturing a wire reference electrode for a biosensor for healthcare according to one embodiment of the present invention may include steps S110, S120, S130, and S140.

[0120]

[0121] S110 step

[0122] Referring to FIGS. 5 and FIGS. 6, the S110 step is a step of preparing a metal wire (110).

[0123] According to one embodiment of the present invention, in step S110, a silver (Ag) wire can be prepared as a metal wire (110).

[0124] The silver (Ag) wire prepared as the metal wire (110) has high conductivity and a high surface area, so signal transmission is very fast and efficient and is also chemically stable.

[0125] At this time, in step S110, the silver (Ag) wire prepared as the metal wire (110) can be washed. In step S110, the silver (Ag) wire prepared as the metal wire (110) can be washed, for example, with isopropyl alcohol.

[0126]

[0127] S120 step

[0128] Referring to FIGS. 7 and FIGS. 8, step S120 is a step of forming a chlorinated layer (120) on the surface of a metal wire (110) by chlorinating the metal wire (110) prepared through step S110.

[0129] According to one embodiment of the present invention, the chlorination layer (120) may be made of silver chloride (AgCl) through chlorination of silver (Ag) as the metal wire (110) is made of silver (Ag) wire.

[0130] To this end, in step S120 above, a chlorination solution can first be prepared (S121).

[0131] At this time, a mixed solution prepared by mixing hydrochloric acid (HCl) and potassium chloride (KCl) may be used as the chlorination solution.

[0132] For example, in step S120 above, a chlorination solution can be prepared by mixing 0.01M hydrochloric acid and 0.1M potassium chloride.

[0133] Next, in step S120, chlorination of the silver (Ag) wire prepared as a metal wire (110) can be carried out in the prepared chlorination solution, that is, a mixed solution of hydrochloric acid and potassium chloride (S122).

[0134] In step S120 above, the chlorination solution is filled into a beaker, and then, according to the 3-electrode system, a silver (Ag) wire prepared as a metal wire (110) is immersed in the beaker filled with the chlorination solution, and then the silver (Ag) wire can be designated as the working electrode (WE).

[0135] In addition, in the above S120 step, for example, a platinum (Pt) electrode can be immersed in a beaker filled with a chlorination solution, and then the platinum (Pt) electrode can be designated as the counter electrode (CE).

[0136] And in the above S120 step, for example, a commercial silver chloride electrode (Ag / AgCl electrode) can be immersed in a beaker filled with a chlorination solution, and then the commercial silver chloride electrode can be designated as the reference electrode (RE).

[0137] Next, in step S120, a silver (Ag) wire designated as the working electrode (WE), a platinum (Pt) electrode designated as the counter electrode (CE), and a commercial silver chloride electrode designated as the reference electrode (RE) can be connected to the electrochemical equipment.

[0138] At this time, in step S120, the chlorination solution may be continuously stirred during the chlorination process to activate electrolyte transfer within the chlorination solution.

[0139] For example, in step S120 above, the chlorination solution can be continuously stirred during the chlorination process at a stirring speed of 300 RPM.

[0140] In the above S120 step, a chlorinated layer (120) made of silver chloride (AgCl) can be formed on the surface of a silver (Ag) wire prepared as a metal wire (110) through chlorination by galvanostatic method.

[0141] At this time, in the above S120 step, chlorination can be carried out by a constant current method for 10 minutes at a current density of 5 mA / cm², for example.

[0142] Accordingly, a chlorinated layer (120) made of silver chloride (AgCl) can be formed on the surface of the silver (Ag) wire prepared as a metal wire (110). That is, the surface of the silver (Ag) wire prepared as a metal wire (110) can be coated with a chlorinated layer (120) made of silver chloride (AgCl).

[0143] In this way, the chlorinated layer (120), i.e., the silver chloride (AgCl) layer formed on the surface of the silver (Ag) wire through chlorination treatment of the silver (Ag) wire prepared as a metal wire (110), can provide a stable reference potential.

[0144] At this time, in the above S120 step, for example, the metal wire (110) having a chlorinated layer (120) formed on its surface can be washed with distilled water and then dried.

[0145]

[0146] S130 step

[0147] Referring to FIGS. 9 and 10, the S130 step involves chloride ions (Cl₂) from a chlorination layer (120) composed of silver chloride (AgCl) formed on the surface of a silver (Ag) wire prepared as a metal wire (110). - The step is to form a first defense layer (130) on the surface of the chlorination layer (120) to physically block the movement of ).

[0148] According to one embodiment of the present invention, in step S130, a polyvinyl butyral resin (PVB) membrane synthesized from polyvinyl alcohol (PVA) and butyraldehyde can be formed as the first protective layer (130).

[0149] To this end, in step S130 above, a polyvinyl butyral solution (PVB solution) can be prepared first (S131).

[0150] In step S130 above, polyvinyl butyral (PVB) powder can be dissolved in methanol to prepare a polyvinyl butyral solution (PVB solution). At this time, in step S130 above, for example, the prepared polyvinyl butyral solution (PVB solution) can be stored at 7°C.

[0151] Next, in step S130, a polyvinyl butyral cocktail (PVB cocktail) can be prepared based on the polyvinyl butyral solution (PVB solution) prepared (S132).

[0152] In step S130 above, sodium chloride (NaCl) and silver nitride (AgNO3) powders can be mixed. Next, in step S130 above, the mixed powder of sodium chloride (NaCl) and silver nitride (AgNO3) can be dissolved in a polyvinyl butyral solution (PVB solution).

[0153] Accordingly, a polyvinyl butyral cocktail based on a polyvinyl butyral solution (PVB solution) can be prepared.

[0154] Next, in step S130, the metal wire (110) having a chlorinated layer (120) formed on its surface can be coated with a polyvinyl butyral cocktail (PVB cocktail) (S133).

[0155] That is, in step S130 above, the surface of the chlorinated layer (120) made of silver chloride (AgCl) formed on the surface of the metal wire (110), i.e., the silver (Ag) wire, can be coated with a polyvinyl butyral cocktail (PVB cocktail).

[0156] At this time, in step S130, a polyvinyl butyral cocktail (PVB cocktail) can be coated on the surface of a chlorinated layer (120) made of silver chloride (AgCl) through dip-coating.

[0157] To this end, in step S130, a metal wire (110) having a chlorinated layer (120) formed on its surface can be immersed in a polyvinyl butyral cocktail (PVB cocktail) for example, about 1 cm and then removed.

[0158] Next, in step S130, a metal wire (110) coated with a polyvinyl butyral cocktail (PVB cocktail) on the outermost surface can be dried and cured (S134).

[0159] In the above S130 step, first, a metal wire (110), i.e., a silver (Ag) wire coated with a polyvinyl butyral cocktail (PVB cocktail) on the outermost surface, can be dried in a dark place for up to 1 hour.

[0160] In the above S130 step, next, a metal wire (110) coated with a polyvinyl butyral cocktail (PVB cocktail) on the outermost surface can be exposed to a UV lamp for up to 10 minutes.

[0161] Through a drying and curing process for a polyvinyl butyral cocktail (PVB cocktail) coated on the surface of a chlorinated layer (120) made of silver chloride (AgCl), the polyvinyl butyral cocktail (PVB cocktail) can be converted into a hard solid film through chemical cross-linking of sodium chloride (NaCl) and silver nitrate (AgNO3) with polyvinyl butyral (PVB).

[0162] That is, during the drying and curing process of the polyvinyl butyral cocktail (PVB cocktail), sodium ions (Na₂C) generated as the sodium chloride (NaCl) and silver nitrate (AgNO₃) dissociate when dissolved in the polyvinyl butyral solution (PVB solution) + ), chloride ions (Cl - ), silver ions (Ag + ) and nitrate ions (NO3 -At least one of the ions forms a cross-link through chemical interaction with the hydroxyl group (-OH) of the polyvinyl butyral (PVB) polymer chain, and through this, the polyvinyl butyral cocktail (PVB cocktail) can be converted into a rigid solid film.

[0163] Here, the polyvinyl butyral (PVB) is sodium ion (Na + ) and silver ions (Ag + It can be cross-linked with ), thereby forming a denser and more robust network. Such cross-linking can contribute to significantly improving the mechanical strength and durability of the solid film.

[0164] In the above S130 step, a solid first protective layer (130) made of polyvinyl butyral (PVB) can be formed on the surface of a chlorinated layer (120) made of silver chloride (AgCl) through the aforementioned series of processes.

[0165] In this way, the first protective layer (130) made of polyvinyl butyral (PVB) is a hydrophobic polymer, and upon curing, forms a dense and rigid network through cross-linking by chemical interaction with ions generated during the process, thereby creating chloride ions (Cl - It can be provided as a barrier that can physically block the movement of ).

[0166] Through this, the first protective layer (130) made of polyvinyl butyral (PVB) can effectively prevent leakage of internal electrolyte while maintaining ion conductivity.

[0167] At this time, the first protective layer (130) made of polyvinyl butyral (PVB) can ensure excellent durability even under repeated mechanical deformation due to the physical properties of polyvinyl butyral (PVB), and accordingly, stable performance can be maintained even during long-term use.

[0168]

[0169] S140 step

[0170] Referring to FIGS. 11 to 13, the S140 step involves chloride ions (Cl₂) from a chlorination layer (120) composed of silver chloride (AgCl) formed on the surface of a silver (Ag) wire prepared as a metal wire (110). - The step is to form a second defense layer (140) on the surface of the first defense layer (130) to physically block the movement of ).

[0171] According to one embodiment of the present invention, in step S140, a second defense layer (140) can be formed with Nafion.

[0172] In the above S140 step, Nafion can be coated on the surface of the first protective layer (130) made of polyvinyl butyral (PVB) through dip-coating.

[0173] In the above S140 step, a metal wire (110) having a first protective layer (130) made of polyvinyl butyral (PVB) formed on its outermost surface can be immersed in, for example, a solution containing 0.5 wt% of Nafion and then removed.

[0174] Next, in step S140, a metal wire (110) coated with Nafion on the outermost surface, i.e., a silver (Ag) wire, can be dried.

[0175] Accordingly, a solid second defense layer (140) made of Nafion can be formed on the surface of a first defense layer (130) made of polyvinyl butyral (PVB).

[0176] Thus, the second defense layer (140) made of Nafion forms the outermost layer of the wire reference electrode (100) for the biosensor being manufactured, and, like the first defense layer (130), chloride ions (Cl) from the chlorination layer (120) made of silver chloride (AgCl) - It can act as a barrier to block the movement of ).

[0177] At this time, the second defense layer (140) made of Nafion is formed from chloride ions (Cl₂) from the chlorination layer (120) made of silver chloride (AgCl). - The movement of ) can be electrostatically blocked.

[0178] That is, the Nafion forming the second defense layer (140) is a negatively charged sulfonic acid group that is a chloride ion (Cl - It can electrostatically repel ). In this case, the hydrophobic polytetrafluoroethylene (PTFE) backbone of Nafion can form a physical barrier.

[0179] In addition, the second protective layer (140) made of Nafion can maintain stability even in high temperature and high humidity environments.

[0180] When the above S140 step is completed, a wire reference electrode (100) for a biosensor according to one embodiment of the present invention can be manufactured.

[0181] At this time, the manufactured wire reference electrode (100) for the biosensor can be stored, for example, in 3M potassium chloride (KCl).

[0182]

[0183] As described above, polyvinyl butyral (PVB) forming the first defense layer (130) and Nafion forming the second defense layer (140) of the biosensor wire reference electrode (100) are chloride ions (Cl - It can block external leakage of ).

[0184] Accordingly, the wire reference electrode (100) for a biosensor manufactured according to one embodiment of the present invention can be provided as an all-solid electrode that does not require a stabilization solution, unlike conventional reference electrodes that require a stabilization solution.

[0185] That is, as the polyvinyl butyral (PVB) forming the first defense layer (130) and the Nafion forming the second defense layer (140) form a double barrier to the outside of the chlorination layer (120) made of silver chloride (AgCl), the physical barrier of the polyvinyl butyral (PVB) forming the first defense layer (130) and the electrostatic barrier of the Nafion forming the second defense layer (140) can exhibit a synergistic effect, and through this, chloride ions (Cl) from the chlorination layer (120) made of silver chloride (AgCl) - The movement of ) can be effectively blocked.

[0186] Accordingly, the wire reference electrode (100) for a biosensor manufactured according to one embodiment of the present invention can have excellent stability and repeatability that operates stably even in extreme environments.

[0187] A method for manufacturing a wire reference electrode for a biosensor according to one embodiment of the present invention has a simple manufacturing process and high reproducibility, enabling mass production and significantly improving economic efficiency.

[0188]

[0189] Example 1

[0190] The prepared silver (Ag) wire was washed with isopropyl alcohol.

[0191] Next, a chlorination solution was prepared by mixing 0.01M hydrochloric acid and 0.1M potassium chloride.

[0192] Next, to proceed with chlorination of the silver (Ag) wire in the prepared chlorination solution, the chlorination solution was filled into a beaker, and the silver (Ag) wire, platinum (Pt) electrode, and commercial silver chloride electrode were immersed in the beaker filled with the chlorination solution. The silver (Ag) wire was designated as the working electrode (WE), the platinum (Pt) electrode as the counter electrode (CE), and the commercial silver chloride electrode as the reference electrode (RE). These were connected to an electrochemical apparatus, and chlorination of the silver (Ag) wire was carried out by constant current at a current density of 5 mA / cm² for 10 minutes. Through this, a silver chloride (AgCl) layer was formed on the surface of the silver (Ag) wire.

[0193] At this time, the chlorination solution was continuously stirred during the chlorination process at a stirring speed of 300 RPM.

[0194] Next, polyvinyl butyral (PVB) powder was dissolved in methanol to prepare a polyvinyl butyral solution, which was stored at 7°C.

[0195] Next, a mixed powder of sodium chloride (NaCl) and silver nitrate (AgNO3) was dissolved in a polyvinyl butyral solution to prepare a polyvinyl butyral cocktail.

[0196] Next, a silver (Ag) wire with a silver chloride (AgCl) layer formed on its surface was immersed in a polyvinyl butyral cocktail (PVB cocktail) for, for example, about 1 cm, and then removed to coat the surface of the silver chloride (AgCl) layer with the polyvinyl butyral cocktail.

[0197] Next, a silver (Ag) wire coated with a polyvinyl butyral cocktail on the outermost layer was dried in a dark place for 1 hour, and then exposed to a UV lamp for 10 minutes to form a solid polyvinyl butyral (PVB) membrane on the surface of the silver chloride (AgCl) layer.

[0198] Next, a silver (Ag) wire with a polyvinyl butyral (PVB) membrane formed on its outermost layer was immersed in a solution containing 0.5 wt% Nafion, removed, and dried to form a solid Nafion layer on the surface of the polyvinyl butyral (PVB) membrane, thereby manufacturing a wire reference electrode.

[0199]

[0200] Figure 14 shows the results of measuring the Open Circuit Potential (OCP) for the electrodes produced at each step of manufacturing the wire reference electrode according to Example 1.

[0201] At this time, a three-electrode system was connected to an electrochemical device using a wire reference electrode manufactured according to Example 1 as the working electrode, a platinum electrode as the counter electrode, and a commercial silver chloride electrode as the reference electrode, and immersed in a 3M potassium chloride (KCl) solution to measure the open-circuit potential of the electrodes produced at each stage.

[0202] Referring to Fig. 14, it was confirmed that the silver (Ag) wire, after being cleaned, exhibited a stable graph shape starting from 3 minutes after measurement. However, the silver (Ag) wire does not possess the characteristics of a reference electrode, which is essential for providing a stable potential through a reversible reaction. This indicates that it is impossible to secure the stability of the electrode because the silver (Ag) wire has difficulty maintaining a constant and highly reproducible potential for a long time, and thus the reliability of the material detection results cannot be guaranteed.

[0203] It was confirmed that the Ag / AgCl wire, which has a silver chloride (AgCl) layer formed through the chlorination process, exhibited a phenomenon where the current dropped sharply with a difference of approximately 80 mV after about 2 minutes of measurement. This is understood to be due to the silver chloride (AgCl) layer formed on the surface of the silver (Ag) wire flaking off, resulting in an unstable potential; it was confirmed that once the silver chloride (AgCl) layer completely disappears, the silver (Ag) wire exhibits a linear potential. Consequently, electrodes composed of Ag / AgCl wires cannot be used as reference electrodes because it is difficult to verify their stability. Therefore, it is necessary to effectively maintain the silver chloride (AgCl) layer.

[0204] In the case of the Ag / AgCl / PVB wire, which has a polyvinyl butyral (PVB) membrane formed on a silver (Ag) wire with a silver chloride (AgCl) layer, it showed the most stable potential pattern among the wires mentioned above, but it was confirmed that the potential increased or decreased unevenly within the 10 mV range.

[0205] On the other hand, the Ag / AgCl / PVB / nafion wire with a Nafion layer formed on the outermost layer showed a standard deviation of about 0.05 mV to 0.37 mV in the open circuit potential (OCP) measurement results and exhibited a linear potential pattern, demonstrating stability that allows it to be used as a reference electrode.

[0206]

[0207] Meanwhile, Figure 15 shows the open-circuit potential measurement results for methanol and acetone-based polyvinyl butyral coated electrodes.

[0208] That is, FIG. 15 is a comparative example 1, in which polyvinyl butyral (PVB) powder is dissolved in a mixed solution of methanol and acetone to prepare a polyvinyl butyral solution, and the polyvinyl butyral (PVB) membrane is formed on a silver (Ag) wire having a silver chloride (AgCl) layer using this polyvinyl butyral solution to measure the open circuit potential of the Ag / AgCl / PVB wire.

[0209] Referring to Fig. 15, the open-circuit potential graph of the Ag / AgCl / PVB wire prepared according to Comparative Example 1 did not exhibit a linear, stabilized shape, but rather showed a tendency for the potential to continuously increase. This was understood to be due to the high volatility of acetone, which prevented the formation of a uniform surface and resulted in an unstable outcome.

[0210] Accordingly, by preparing a polyvinyl butyral (PVB) solution using only methanol and forming a polyvinyl butyral (PVB) membrane, the range of potential change was reduced to within approximately 16 mV.

[0211] Afterwards, the drying time was optimized by dip-coating with a polyvinyl butyral cocktail.

[0212] Figure 16 shows the open-circuit potential measurement results for an undried electrode that underwent UV curing immediately without drying after polyvinyl butyral coating.

[0213] Referring to Figure 16, it was found that in the case of an undried electrode, the solvent in the polyvinyl butyral cocktail remained on the electrode surface without being sufficiently dried, and the polyvinyl butyral membrane was not formed uniformly, which is the cause of the repeated uneven increase and decrease in potential.

[0214] To improve this, a drying time of 10 minutes was allowed after dip-coating, followed by UV curing for 30 minutes.

[0215] Figure 17 shows the results of open-circuit potential measurements for a UV photocurable electrode for 30 minutes after drying the coated polyvinyl butyral.

[0216] Referring to Fig. 17, the open-circuit potential change was stabilized within 5 mV through drying. Nevertheless, it was confirmed that the potential did not exhibit linear stability and repeatedly increased and decreased. This was determined to be due to the fact that the polymer structure of polyvinyl butyral (PVB) was damaged as excessive curing occurred through UV curing beyond what was necessary, in addition to the discoloration of the electrode color during UV curing.

[0217] Figure 18 shows the results of measuring the open-circuit potential of coated polyvinyl butyral on a UV photocuring electrode for 10 minutes.

[0218] Referring to Fig. 18, it was confirmed that when the UV curing time was reduced to 10 minutes, the open circuit potential showed stability.

[0219] At this time, the open-circuit potential evaluation results for two Ag / AgCl / PVB / nafion wire electrodes showed that the potential change was measured to be 0.039 mV and 0.0283 mV, respectively, indicating that it is a sufficiently stable level to be used as a reference electrode in electrochemical measurements.

[0220] As such, the Ag / AgCl / PVB / nafion wire electrode, prepared by UV-curing coated polyvinyl butyral for 10 minutes, maintained a constant potential for a long period while minimizing the influence of external environmental changes such as temperature, humidity, and solution composition. Through this, it was confirmed that the Ag / AgCl / PVB / nafion wire electrode, prepared by UV-curing polyvinyl butyral for 10 minutes, can provide repetitive and highly reliable electrochemical data and is suitable as a reference electrode for a sensor platform.

[0221] In particular, this stability demonstrates superior results when compared to conventional silver chloride (AgCl) electrodes or electrodes coated only with polyvinyl butyral (PVB), thereby proving the effectiveness of the wire reference electrode manufacturing technology according to the present invention.

[0222] As such, the wire electrode manufactured according to Example 1, namely the Ag / AgCl / PVB / nafion wire electrode, can be applied as a platform for a sensor that precisely measures various biomarkers such as glucose and cholesterol, and can provide high reproducibility and reliability as a reference electrode.

[0223]

[0224] FIGS. 19 and 20 are scanning electron microscope images of a cross-section of a wire reference electrode manufactured according to Example 1, and FIGS. 21 to 25 are the results of analyzing the wire reference electrode manufactured according to Example 1 through energy dispersive spectroscopy (EDS).

[0225] Referring to FIGS. 19 and 20, a silver chloride (AgCl) layer formed around a silver (Ag) wire, a polyvinyl butyral (PVB) membrane surrounding this layer, and a Nafion layer formed on the outermost surface were observed.

[0226] Referring to Fig. 21, silver (Ag) distributed in the silver (Ag) wire at the center of the wire reference electrode and the silver chloride (AgCl) layer was confirmed.

[0227] Furthermore, referring to Fig. 22, chlorine (Cl) forming a silver chloride (AgCl) layer and a polyvinyl butyral (PVB) membrane was also identified.

[0228] Furthermore, referring to FIG. 23, since polyvinyl butyral (PVB) contains not only chlorine (Cl) but also hydroxyl groups (-OH) and ether groups (-O-), oxygen (O) inside acts as a major constituent element during the formation of the polyvinyl butyral (PVB) membrane. Considering this, it was determined that the polyvinyl butyral (PVB) membrane is well formed through the distribution of oxygen (O) in the polyvinyl butyral (PVB) membrane.

[0229] Furthermore, referring to Fig. 24, it was confirmed that a Nafion layer was formed on the outermost surface through fluorine (F), a key constituent element contained in large quantities in the Nafion polymer backbone (-CF2-CF2-) structure.

[0230] Meanwhile, referring to FIG. 25, the presence of silver (Ag), chlorine (Cl), oxygen (O), and fluorine (F) elements can be confirmed. At this time, since the wire reference electrode prepared according to Example 1 was stored in 3M potassium chloride (KCl), some potassium (K) was detected.

[0231] In summary, it was confirmed that the target layer was formed well at each step of manufacturing the wire reference electrode according to Example 1.

[0232]

[0233] FIGS. 26 and 27 are results of measuring the concentration of glucose using chronoamperometry (CA) with a glucose sensor fabricated using a wire reference electrode manufactured according to Example 1.

[0234] Referring to FIG. 26, when glucose at concentrations of 0.5 mM, 1 mM, 2 mM, 3 mM, 4 mM, and 5 mM was provided at 50-second intervals from 0 to 400 seconds, it was confirmed that the current density increased stepwise as the sensor detected glucose at each concentration. This indicates that the wire reference electrode manufactured according to Example 1 is functioning properly.

[0235] In addition, referring to Fig. 27, it was confirmed that the current density increased in a constant linear fashion according to glucose concentrations of 0.5 mM, 1 mM, 2 mM, 3 mM, 4 mM, and 5 mM. This means that the wire reference electrode prepared according to Example 1 has been stabilized and can actually be used as a wire reference electrode.

[0236] These results show that the wire reference electrode manufactured according to Example 1 can be used as a platform for a sensor that detects substances such as glucose, cholesterol, and electrolytes.

[0237]

[0238] Although the present invention has been described in detail using preferred embodiments, the scope of the invention is not limited to specific embodiments and should be interpreted by the appended claims. Furthermore, those skilled in the art will understand that many modifications and variations are possible without departing from the scope of the invention.

Claims

1. Metal wire; A chlorinated layer provided on the surface of the metal wire and formed through chlorination of the metal constituting the metal wire; It is provided on the surface of the chlorination layer, and chloride ions (Cl) from the chlorination layer - A first defense layer that physically blocks the movement of ); and It is provided on the surface of the first protective layer, and the chloride ions (Cl - A wire reference electrode for a biosensor for healthcare, comprising a second defense layer that electrostatically blocks the movement of ).

2. In Paragraph 1, The above metal wire is made of silver (Ag), and A wire reference electrode for a biosensor for healthcare, wherein the above-mentioned chlorinated layer is composed of silver chloride (AgCl).

3. In Paragraph 2, The first protective layer above is made of polyvinyl butyral (PVB), and The above second protective layer is a wire reference electrode for a biosensor for healthcare, composed of Nafion.

4. In Paragraph 1, Wire reference electrode for a biosensor for healthcare, forming an all-solid-state electrode.

5. Wire reference electrode; and A sensor part provided on the above-mentioned wire reference electrode and made of a material that responds to a target biomaterial; comprising, The above wire reference electrode is, Metal wire; A chlorinated layer provided on the surface of the metal wire and formed through chlorination of the metal constituting the metal wire; It is provided on the surface of the chlorination layer, and chloride ions (Cl) from the chlorination layer - A first defense layer that physically blocks the movement of ); and It is provided on the surface of the first protective layer, and the chloride ions (Cl - A healthcare biosensor comprising a second defense layer that electrostatically blocks the movement of ).

6. Preparation step for preparing metal wire; A chlorination layer formation step of forming a chlorination layer on the surface of the metal wire through chlorination of the metal wire prepared above; Chloride ions (Cl from the chlorination layer formed on the surface of the metal wire) - A first protective layer forming step of forming a first protective layer on the surface of the chlorinated layer to physically block the movement of ); and The above chloride ion (Cl - A method for manufacturing a wire reference electrode for a biosensor for healthcare, comprising: a second defense layer forming step of forming a second defense layer on the surface of the first defense layer to electrostatically block the movement of ).

7. In Paragraph 6, In the above preparation step, a metal wire made of silver (Ag) is prepared, and A method for manufacturing a wire reference electrode for a biosensor for healthcare, wherein in the step of forming the chlorination layer, a chlorination layer composed of silver chloride (AgCl) is formed on the surface of the metal wire.

8. In Paragraph 7, A method for manufacturing a wire reference electrode for a biosensor for healthcare, wherein the first protective layer is formed with polyvinyl butyral (PVB) in the first protective layer formation step.

9. In Paragraph 8, The above first defense layer formation step is, Process for preparing a polyvinyl butyral (PVB) solution; A process of preparing a polyvinyl butyral cocktail by dissolving a mixed powder of sodium chloride (NaCl) and silver nitrate (AgNO3) in the polyvinyl butyral (PVB) solution; A process of coating a metal wire having a chlorinated layer formed on its surface with the manufactured polyvinyl butyral cocktail; A method for manufacturing a wire reference electrode for a biosensor for healthcare, comprising the process of drying and curing a metal wire coated with a polyvinyl butyral cocktail on the outermost surface so that a first protective layer made of polyvinyl butyral (PVB) is formed on the surface of the chlorinated layer.

10. In Paragraph 9, A method for manufacturing a wire reference electrode for a biosensor for healthcare, wherein in the above drying and curing process, a metal wire coated with the polyvinyl butyral cocktail on the outermost surface is dried for up to 1 hour, and then exposed to a UV lamp for up to 10 minutes.

11. In Paragraph 10, In the above drying and curing process, sodium ions (Na₂C₃) generated when the sodium chloride (NaCl) and silver nitrate (AgNO₃) dissociate upon dissolving in the polyvinyl butyral (PVB) solution + ), chloride ions (Cl - ), silver ions (Ag + ) and nitrate ions (NO3 - A method for manufacturing a wire reference electrode for a biosensor for healthcare, wherein at least one of the ions forms a cross-link through chemical interaction with the hydroxyl group (-OH) of a polyvinyl butyral (PVB) polymer chain.

12. In Paragraph 8, A method for manufacturing a wire reference electrode for a biosensor for healthcare, wherein the second defense layer is formed with Nafion in the second defense layer formation step.

Images