CN112798664A - Electrochemical test paper capable of continuously detecting substrate - Google Patents
Electrochemical test paper capable of continuously detecting substrate Download PDFInfo
- Publication number
- CN112798664A CN112798664A CN202011616816.9A CN202011616816A CN112798664A CN 112798664 A CN112798664 A CN 112798664A CN 202011616816 A CN202011616816 A CN 202011616816A CN 112798664 A CN112798664 A CN 112798664A
- Authority
- CN
- China
- Prior art keywords
- test paper
- electrochemical test
- layer
- substrate
- continuously detecting
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3271—Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
- G01N27/3272—Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Biophysics (AREA)
- Hematology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The application relates to the technical field of electrochemistry, provides an electrochemistry test paper that can detect substrate in succession, electrochemistry test paper include the substrate, set up in reference electrode, counter electrode and working electrode on substrate surface, with the reference electrode utmost point the counter electrode with the circuit that the working electrode links to each other to and set up the reaction layer on the working electrode, its characterized in that, the reaction layer includes the conducting substrate layer and sets up the functional layer on conducting substrate layer surface, the functional layer includes porous nanometer electro-catalysis material, at least one with detect the enzyme of material reaction, polymer binder and cross-linking agent, the electrochemistry test paper that obtains including the reaction layer can detect high-content substrate in succession, and repeated the use, the range of application is wider.
Description
Technical Field
The application belongs to the technical field of electrochemical test paper, and particularly relates to electrochemical test paper capable of continuously detecting a substrate.
Background
The use of electrochemical strips has become a popular technique for medical or biochemical testing. Conventional electrochemical test strips have two electrodes, and after a sample is injected into a reaction region of the test strip, the electrical characteristics of the sample are measured by the two electrodes.
At present, some indexes need to be monitored and measured every day by patients, for example, hypertension patients need to measure blood pressure every day, hyperglycemia patients need to measure blood sugar every day, hyperlipidemia patients need to measure blood fat every day, patients suffering from heart diseases need to measure heart rate every day, and the like. Therefore, some household portable testers, such as blood glucose tester, blood lipid tester, uric acid tester, lactic acid tester and corresponding electrochemical test paper, appear on the market.
The existing electrochemical test paper for detecting the human body marker is generally disposable, and because part of substances in a component material or a reagent of a working electrode of the test paper are easy to lose, the signal attenuation is fast when the electrochemical test paper is repeatedly used, the test paper cannot meet the requirement of continuous detection on occasions needing repeated or continuous detection, such as a sensor for measuring sweat, and the existing test paper cannot meet the requirement. Meanwhile, the content of some metabolites in sweat is high, while the detection upper limit of a sensor which can continuously detect in literature reports is low.
Disclosure of Invention
The application aims to provide an electrochemical test paper capable of continuously detecting a substrate, and aims to solve the problems that the electrochemical test paper in the prior art cannot continuously detect and the upper limit of detection is low.
In order to achieve the purpose of the application, the technical scheme adopted by the application is as follows:
in a first aspect, the present application provides an electrochemical test paper capable of continuously detecting a substrate, the electrochemical test paper includes a substrate, a reference electrode, a counter electrode, a working electrode disposed on a surface of the substrate, a circuit connected to the reference electrode, the counter electrode, and the working electrode, and a reaction layer disposed on the working electrode, the reaction layer includes a conductive substrate layer and a functional layer disposed on a surface of the conductive substrate layer, and the functional layer includes a porous nano electro-catalytic material, at least one enzyme reacting with a substance to be detected, a polymer binder, and a cross-linking agent.
The electrochemical test paper capable of continuously detecting the substrate provided by the first aspect of the application comprises a structure of common electrochemical test paper, wherein a reaction layer comprises a conductive substrate layer and a functional layer arranged on the surface of the conductive substrate layer, and the functional layer comprises a porous nano electro-catalytic material, at least one enzyme reacting with a detected substance, a high-molecular adhesive and a cross-linking agent; the conductive substrate layer is used as a carrier of the reaction layer and can carry various substances; the porous nano electro-catalytic material loaded with the surfactant or the silane coupling agent can control the diffusion of components in a substance to be detected, provide sufficient oxygen and control the diffusion rate of the oxygen based on a porous structure formed on the surface, thereby realizing high-content detection; meanwhile, the material has stable property, and can realize continuous detection for multiple uses; the polymer adhesive is provided, so that the porous nano electro-catalysis material and the enzyme can be ensured to be fixed on the conductive substrate, the diffusion effect can be controlled, and the high-content detection can be realized; the cross-linking agent is provided, the cross-linking agent and at least one enzyme which reacts with the detected substance form an enzyme-cross-linking agent intermediate, when detection is carried out, the detected substance can quickly react with the enzyme-cross-linking agent intermediate, and the detection efficiency is improved; the prepared electrochemical test paper comprising the reaction layer can continuously detect the substrate with the concentration content as high as 50mM, the upper limit of detection is high, the electrochemical test paper can continuously detect the substrate, the detection range is wide, the electrochemical test paper can be repeatedly used, and the application range is wider.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the embodiments or the prior art descriptions will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 is a block diagram of an electrochemical test strip provided in an embodiment of the present application.
Fig. 2 is a structural diagram of a reaction layer of the electrochemical test strip provided in the example of the present application.
FIG. 3 is a graph of data for continuous lactate determination provided in the examples of the present application.
Detailed Description
In order to make the technical problems, technical solutions and advantageous effects to be solved by the present application more clearly apparent, the present application is further described in detail below with reference to the embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the present application and are not intended to limit the present application.
In this application, the term "and/or" describes an association relationship of associated objects, meaning that there may be three relationships, e.g., a and/or B, which may mean: a is present alone, A and B are present simultaneously, and B is present alone. Wherein A and B can be singular or plural. The character "/" generally indicates that the former and latter associated objects are in an "or" relationship.
In the present application, "at least one" means one or more, "a plurality" means two or more. "at least one of the following" or similar expressions refer to any combination of these items, including any combination of the singular or plural items. For example, "at least one (a), b, or c", or "at least one (a), b, and c", may each represent: a, b, c, a-b (i.e., a and b), a-c, b-c, or a-b-c, wherein a, b, and c may be single or plural, respectively.
It should be understood that, in various embodiments of the present application, the sequence numbers of the above-mentioned processes do not mean the execution sequence, some or all of the steps may be executed in parallel or executed sequentially, and the execution sequence of each process should be determined by its function and inherent logic, and should not constitute any limitation to the implementation process of the embodiments of the present application.
The terminology used in the embodiments of the present application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. As used in the examples of this application and the appended claims, the singular forms "a", "an", and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.
The weight of the related components mentioned in the description of the embodiments of the present application may not only refer to the specific content of each component, but also represent the proportional relationship of the weight among the components, and therefore, the content of the related components is scaled up or down within the scope disclosed in the description of the embodiments of the present application as long as it is scaled up or down according to the description of the embodiments of the present application. Specifically, the mass described in the specification of the embodiments of the present application may be a mass unit known in the chemical industry field such as μ g, mg, g, kg, etc.
The terms "first" and "second" are used for descriptive purposes only and are used for distinguishing purposes such as substances from one another, and are not to be construed as indicating or implying relative importance or implying any number of technical features indicated. For example, a first XX may also be referred to as a second XX, and similarly, a second XX may also be referred to as a first XX, without departing from the scope of embodiments of the present application. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature.
In a first aspect, the present embodiment provides an electrochemical test strip capable of continuously detecting a substrate, as shown in fig. 1, the electrochemical test strip includes a substrate 1, a reference electrode 2, a counter electrode 3, and a working electrode 4 disposed on a surface of the substrate 1, a circuit 5 connected to the reference electrode 2, the counter electrode 3, and the working electrode 4, and a reaction layer 6 disposed on the working electrode 4; as shown in fig. 2, the reaction layer 6 includes a conductive substrate layer 7 and a functional layer 8 disposed on the surface of the conductive substrate layer 7, and the functional layer 8 includes a porous nano electro-catalytic material loaded with a surfactant or a silane coupling agent, at least one enzyme reacting with a substance to be detected, a polymer binder, and a cross-linking agent.
The electrochemical test paper capable of continuously detecting the substrate provided by the first aspect of the application comprises a structure of a common electrochemical test paper, wherein the reaction layer 6 comprises a conductive substrate layer 7 and a functional layer 8 arranged on the conductive substrate layer 7, and the functional layer 8 comprises a porous nano electro-catalytic material loaded with a surfactant or a silane coupling agent, at least one enzyme reacting with a detected substance, a high molecular adhesive and a cross-linking agent. The conductive substrate layer 7 is used as a carrier of the reaction layer 6 and can carry various substances; the functional layer 8 comprises a porous nano electro-catalytic material loaded with a surfactant or a silane coupling agent, and can control the diffusion of components in a substance to be detected, provide sufficient oxygen, control the diffusion rate of the oxygen and realize high-content detection based on a porous structure formed on the surface; meanwhile, the material has stable property, and can realize continuous detection for multiple uses; the polymer adhesive is provided, so that the porous nano electro-catalysis material and the enzyme can be ensured to be fixed on the conductive substrate, the diffusion effect can be controlled, and the high-content detection can be realized; the cross-linking agent is provided, the cross-linking agent and at least one enzyme which reacts with the detected substance form an enzyme-cross-linking agent intermediate, when detection is carried out, the detected substance can quickly react with the enzyme-cross-linking agent intermediate, and the detection efficiency is improved; the prepared electrochemical test paper comprising the reaction layer can continuously detect a substrate with the concentration as high as 50mM, can be repeatedly used, and has a wider application range.
The materials of the reference electrode 2, the counter electrode 3 and the working electrode 4 may be selected from various materials conventionally used, and are not particularly limited. In addition, the arrangement of the reference electrode 2, the counter electrode 3, the working electrode 4 and the connected circuit is not required, and the electrochemical test paper only needs to be ensured to work normally.
Specifically, the reaction layer is arranged on the working electrode, and the arrangement of the reaction layer can influence the detection performance of the electrochemical test paper and improve the detection effect.
Further, the reaction layer 6 comprises a conductive substrate layer 7 and a functional layer 8 arranged on the surface of the conductive substrate layer 7, and the functional layer 8 comprises a porous nano electro-catalytic material, at least one enzyme reacting with the detected substance, a high molecular adhesive and a cross-linking agent.
The conductive substrate layer 7 is arranged to provide a carrier for other materials, so that each component can be loaded on the surface of the conductive substrate layer to improve the detection performance of the object to be detected.
In some embodiments, the material of the conductive substrate layer 7 is selected from any one of nickel foam, noble metal, carbon fiber, carbon tube, graphite, and glassy carbon. The materials are selected as the conductive substrate layer, on one hand, the materials are stable in property and difficult to generate chemical reaction, the substrate layer can be ensured not to react with each loaded component, the reaction sensitivity is controlled, and the obtained electrochemical test paper is ensured to be easy to store and not to react with substances in the air, so that the use is influenced; on the other hand, the electric signals can be increased, and the test sensitivity and accuracy are improved.
In some embodiments, the thickness of the conductive substrate layer 7 is 0.1-1000 micrometers, and the thickness can be selected according to specific requirements, so that normal use can be ensured. In the embodiment of the invention, the thickness of the conductive substrate layer 7 is 20-100 micrometers, and the thickness of the conductive substrate layer is controlled to be 20-100 micrometers, so that the electrochemical test paper is more suitable for preparing electrochemical test paper.
Further, the reaction layer 6 includes a functional layer 8 disposed on the surface of the conductive substrate layer 7, the functional layer 8 includes a porous nano electro-catalytic material, at least one enzyme reacting with the substance to be detected, a polymer binder and a cross-linking agent, and the functional layer provides various functional materials for reaction.
In some embodiments, the thickness of the functional layer 8 is 1 to 500 microns, and controlling the thickness of the functional layer further controls the effectiveness of the detection. In the preferred embodiment of the invention, the thickness of the functional layer 8 is 10-100 microns, if the thickness of the functional layer is too thin, the electrocatalytic activity is insufficient, and the effect of continuously detecting the high-content object to be detected for many times cannot be realized; if the thickness of the functional layer is too thick, the linearity of the detection effect is poor, and the accuracy is affected. In the preferred embodiment of the invention, the thickness of the functional layer 8 is 30-50 micrometers, and the thickness of the functional layer is controlled to be 30-50 micrometers, so that the obtained electrochemical test paper can be ensured to realize continuous and repeated detection of high-content objects to be detected, and the electrochemical test paper has high accuracy and good detection effect.
Further, the functional layer 8 comprises a porous nano-electrocatalytic material. The porous nano electro-catalytic material is provided, and based on a porous structure formed on the surface of the porous nano electro-catalytic material, the diffusion of components in a substance to be detected can be controlled, sufficient oxygen is provided, the diffusion rate of the oxygen is controlled, and high-content detection is realized; meanwhile, the material has stable property, and can realize continuous detection for multiple uses.
In some embodiments, the porous nanoelectrocatalytic material is selected from at least one of mesoporous phosphonate materials, metal organic framework materials, covalent organic framework materials, carbonized derivatives of metal organic framework materials, and carbonized derivatives of covalent organic framework materials. The nano-scale electrocatalytic material with pores is selected, so that on one hand, an electrochemical reaction can be realized, on the other hand, the material with a pore structure is introduced into the electrochemical test paper, a certain amount of oxygen can be provided for the material, the diffusion rate of the oxygen is controlled, and high-content detection is realized; meanwhile, the provided material has stable property and can be continuously realized for many times.
In some embodiments, the mesoporous phosphonate material is selected from at least one of a mesoporous organic phosphonate material, a mesoporous inorganic phosphonate material, a mesoporous organic phosphonate supported noble metal material, a mesoporous organic phosphonate supported transition metal material. The selected mesoporous phosphonate material has a mesoporous aperture of 2-50 nm, and the diffusion rate of oxygen can be properly controlled by controlling the aperture, so that the detection sensitivity is prevented from being low.
In some embodiments, the surface of the porous nanoelectrocatalytic material is loaded with at least one of a surfactant, a silane coupling agent. The surfactant or the silane coupling agent is attached, and the surfactant or the silane coupling agent contains a certain amount of polar groups, so that the adsorption capacity is strong, and the detection of high-content substances can be realized.
In some embodiments, the surface of the porous nanoelectrocatalytic material is loaded with a surfactant. In other embodiments, the surface of the porous nanoelectrocatalytic material is loaded with a silane coupling agent. In still other embodiments, the surface of the porous nanoelectrocatalytic material is loaded with a surfactant and a silane coupling agent.
In some embodiments, the surfactant is selected from at least one of a carboxylate surfactant, a sulfonate surfactant, a sulfate surfactant, and a phosphate surfactant.
In some embodiments, the silane coupling agent is selected from at least one of chloro-silane, methoxy-silane, ethoxy-silane, acetoxy-silane, methoxy-ethoxy-silane.
Further, the functional layer 8 further includes at least one enzyme that reacts with the substance to be detected, and at least one enzyme that reacts with the substance to be detected is provided according to the substance to be detected, and the enzyme is provided to perform selective detection of a specific substance and determine the detection object of the electrochemical test paper used. In some embodiments, two or more different enzymes that react with the same substance to be detected may be used to enhance the detection.
In a preferred embodiment of the present invention, when glucose is detected, at least one of glucose oxidase and glucose dehydrogenase is added; when detecting uric acid, at least one of urate oxidase and urate dehydrogenase is added; when lactic acid is detected, at least one of lactate oxidase and lactate dehydrogenase is added. The type of the corresponding enzyme is set according to the type of substance to be detected.
In some embodiments, the enzyme is added in an amount of 0.01 to 100 units. The addition of the enzyme is controlled to be more, so that the continuous and repeated determination of the high-content object to be detected can be ensured; if the addition amount of the enzyme is too small, a high-content object to be detected cannot be detected, so that the detection effect is poor, and the frequency of repeated use is also influenced. In the embodiment of the invention, the addition amount of the enzyme is 0.2-5 units.
Further, the functional layer 8 comprises a polymer adhesive, and the polymer adhesive is provided, so that the composite material of the functional layer can be ensured to be fixed on the conductive substrate layer 7, the functional layer 8 is ensured not to fall off easily, and the electrochemical test paper is favorable for long-term storage and repeated use; moreover, the diffusion effect can be controlled, the determination of high-content substances can be realized, and the method can be applied to the continuous monitoring of substances to be detected.
In some embodiments, the polymeric binder is selected from at least one of perfluorosulfonic acid type polymers, chitosan, guar gum, gum arabic, polyurethane, polyvinyl butyral, cellulose, nanocellulose.
In some embodiments, the mass ratio of the polymer binder to the porous nano electro-catalytic material is (0.1-3): 1, the addition amount of the high-molecular adhesive is controlled, so that the porous nano electro-catalytic material and the enzyme can be ensured to be fixed on the conductive substrate, and the reaction is ensured. If the amount of the polymeric binder is too large, the electrocatalytic activity of the porous nanoelectrocatalytic material is affected, and if the amount is too small, the porous nanoelectrocatalytic material cannot be stably fixed on the surface of the conductive substrate, which affects the effect of continuous use.
Further, the functional layer 8 comprises a cross-linking agent, the cross-linking agent and at least one enzyme which reacts with the detected substance form an enzyme-cross-linking agent intermediate, and when the detection is carried out, the detected substance can quickly react with the enzyme-cross-linking agent intermediate, so that the detection efficiency is improved; the prepared electrochemical test paper comprising the reaction layer 6 can continuously detect high-content substrates, can be repeatedly used, and has a wider application range.
In some embodiments, the crosslinking agent is selected from aldehydes or carbodiimides. Further, the aldehydes are selected from glutaraldehyde; the carbodiimide is selected from 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride or N-hydroxysuccinimide.
In some embodiments, the addition amount of the cross-linking agent aqueous solution in each functional layer 8 is 1 to 50 μ l, and the addition amount of the cross-linking agent aqueous solution is controlled according to the thickness of the functional layer 8 and the actual situation. Furthermore, the cross-linking agent aqueous solution contains 0.5-5% of cross-linking agent by mass percentage.
Wherein, the electrochemical test paper has wide linear range, and the linear range capable of detecting is 0.005-50 mM.
The following description will be given with reference to specific examples.
Example 1
Electrochemical test paper for detecting lactic acid
An electrochemical test paper capable of continuously detecting high-content lactic acid comprises a substrate 1, a reference electrode 2, a counter electrode 3 and a working electrode 4 which are arranged on the surface of the substrate 1, a circuit 5 connected with the reference electrode 2, the counter electrode 3 and the working electrode 4, and a reaction layer 6 arranged on the working electrode 4; the reaction layer 6 comprises a conductive substrate layer 7 and a functional layer 8 arranged on the surface of the conductive substrate layer 7, wherein the functional layer 8 comprises a porous nano electro-catalytic material, at least one enzyme reacting with a substance to be detected, a high-molecular adhesive and a cross-linking agent;
wherein the material of the conductive substrate layer is selected from foamed nickel with the thickness of 5 microns; the thickness of the functional layer is 10 microns, and the porous nano electro-catalytic material is selected from a mesoporous phosphonate material of which the surface is loaded with a surfactant fatty acid salt surfactant; the enzyme is selected from lactate oxidase with the addition amount of 50 units, the high molecular adhesive is selected from chitosan, and the mass ratio of the high molecular adhesive to the porous nano electro-catalytic material is 0.5: 1; the cross-linking agent is selected from glutaraldehyde.
Example 2
Electrochemical test paper for detecting glucose
An electrochemical test paper capable of continuously detecting high-content glucose comprises a substrate 1, a reference electrode 2, a counter electrode 3 and a working electrode 4 which are arranged on the surface of the substrate 1, a circuit 5 connected with the reference electrode 2, the counter electrode 3 and the working electrode 4, and a reaction layer 6 arranged on the working electrode 4; the reaction layer 6 comprises a conductive substrate layer 7 and a functional layer 8 arranged on the surface of the conductive substrate layer 7, wherein the functional layer 8 comprises a porous nano electro-catalytic material, at least one enzyme reacting with a substance to be detected, a high-molecular adhesive and a cross-linking agent;
wherein the material of the conductive substrate layer is selected from graphite with the thickness of 10 microns; the thickness of the functional layer is 50 microns, and the porous nano electro-catalytic material is selected from a metal organic framework material of which the surface is loaded with a surfactant alkyl sulfonate surfactant; the enzyme is selected from glucose oxidase with the addition of 80 units, the high-molecular adhesive is selected from perfluorosulfonic acid polymer solution (Nafion), and the mass ratio of the high-molecular adhesive to the porous nano electro-catalytic material is 1.5: 1; the cross-linking agent is selected from glutaraldehyde.
Comparative example 1
Commercially available ordinary lactic acid electrochemical test paper
Comparative example 2
Commercially available ordinary glucose electrochemical test paper
And (3) performance testing:
the concentration of lactic acid in the sweat after exercise is high and can reach 30-50 mM, so that the sweat after exercise is used as a sample to be tested, and the embodiments 1-2 and the comparative examples 1-2 are used as electrochemical test paper for testing.
And (4) analyzing results:
the electrochemical test paper for detecting lactic acid provided in embodiment 1 can accurately detect lactic acid substances in sweat, and can detect lactic acid with a high content (the upper limit of detection is 50mM) or more; the commercially available ordinary lactic acid electrochemical test paper provided in comparative example 1 has an upper limit of detection of 12mM and can be used only once. As shown in FIG. 3, a continuous lactic acid detection data chart shows that the electrochemical test paper can continuously detect lactic acid. The electrochemical test paper for detecting glucose provided by the embodiment 2 can accurately detect glucose substances in the fermentation process, and can realize the determination of glucose with high content (the upper detection limit is 50mM) or more; the commercial ordinary glucose electrochemical test paper provided in comparative example 1 has an upper detection limit of 30mM, but can be used only once, and cannot be used for continuous measurement.
In summary, the electrochemical test paper comprises a structure of a common electrochemical test paper, wherein the reaction layer comprises a conductive substrate layer, a porous nano electro-catalytic material arranged on the conductive substrate layer, at least one enzyme reacting with a substance to be detected, a high molecular adhesive and a cross-linking agent; the conductive substrate layer is used as a carrier of the reaction layer and can carry various substances; the porous nano electro-catalytic material is based on a porous structure formed on the surface, can control the diffusion of components in a substance to be detected, provides sufficient oxygen, controls the diffusion rate of the oxygen and realizes high-content detection; meanwhile, the material has stable property, and can realize continuous detection for multiple uses; the polymer adhesive is provided, so that the porous nano electro-catalysis material and the enzyme can be ensured to be fixed on the conductive substrate, the diffusion effect can be controlled, and the high-content detection can be realized; the cross-linking agent is provided, the cross-linking agent and at least one enzyme which reacts with the detected substance form an enzyme-cross-linking agent intermediate, when detection is carried out, the detected substance can quickly react with the enzyme-cross-linking agent intermediate, and the detection efficiency is improved; the prepared electrochemical test paper comprising the reaction layer can continuously detect high-content substrates, can be repeatedly used, and has a wider application range.
The present invention is not intended to be limited to the particular embodiments shown and described, but is to be accorded the widest scope consistent with the principles and novel features herein disclosed.
Claims (10)
1. The electrochemical test paper comprises a substrate, a reference electrode, a counter electrode and a working electrode which are arranged on the surface of the substrate, a circuit connected with the reference electrode, the counter electrode and the working electrode, and a reaction layer arranged on the working electrode, and is characterized in that the reaction layer comprises a conductive substrate layer and a functional layer arranged on the surface of the conductive substrate layer, and the functional layer comprises a porous nano electro-catalytic material, at least one enzyme reacting with a detected substance, a high-molecular adhesive and a cross-linking agent.
2. The electrochemical strip for continuous detection of a substrate according to claim 1, wherein the material of the conductive substrate layer is selected from any one of nickel foam, noble metal, carbon fiber, carbon tube, graphite and glassy carbon.
3. The electrochemical test strip capable of continuously detecting a substrate according to claim 1, wherein the porous nano electro-catalytic material is at least one selected from mesoporous phosphonate materials, metal organic framework materials, covalent organic framework materials, carbonized derivatives of metal organic framework materials, and carbonized derivatives of covalent organic framework materials.
4. The electrochemical test paper capable of continuously detecting a substrate according to any one of claims 1 to 3, wherein at least one of a surfactant and a silane coupling agent is loaded on the surface of the porous nano-electrocatalytic material.
5. The electrochemical strip for continuously detecting a substrate according to claim 4, wherein the surfactant is at least one selected from the group consisting of a carboxylate surfactant, a sulfonate surfactant, a sulfate surfactant, and a phosphate surfactant; and/or the presence of a gas in the gas,
the silane coupling agent is at least one selected from chloro silane, methoxy silane, ethoxy silane, acetoxy silane and methoxy ethoxy silane.
6. The electrochemical test paper capable of continuously detecting a substrate according to any one of claims 1 to 3, wherein the polymer binder is at least one selected from perfluorosulfonic acid polymers, chitosan, guar gum, gum arabic, polyurethane, polyvinyl butyral, cellulose, and nanocellulose.
7. The electrochemical test paper capable of continuously detecting a substrate according to any one of claims 1 to 3, wherein the crosslinking agent is selected from aldehydes or carbodiimides.
8. The electrochemical test paper capable of continuously detecting a substrate according to any one of claims 1 to 3, wherein the thickness of the conductive substrate layer is 0.1 to 1000 μm; and/or the presence of a gas in the gas,
the thickness of the functional layer is 1-500 microns.
9. The electrochemical test paper for continuous detection of a substrate according to any one of claims 1 to 3, wherein the enzyme is added in an amount of 0.01 to 100 units.
10. The electrochemical test paper capable of continuously detecting a substrate according to any one of claims 1 to 3, wherein the mass ratio of the polymer binder to the porous nano electro-catalytic material is (0.1 to 3): 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011616816.9A CN112798664B (en) | 2020-12-30 | 2020-12-30 | Electrochemical test paper capable of continuously detecting substrate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011616816.9A CN112798664B (en) | 2020-12-30 | 2020-12-30 | Electrochemical test paper capable of continuously detecting substrate |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112798664A true CN112798664A (en) | 2021-05-14 |
| CN112798664B CN112798664B (en) | 2023-03-10 |
Family
ID=75804743
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011616816.9A Active CN112798664B (en) | 2020-12-30 | 2020-12-30 | Electrochemical test paper capable of continuously detecting substrate |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112798664B (en) |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1737560A (en) * | 2005-09-02 | 2006-02-22 | 天津大学 | Static electricity spinning prepared fixed enzymic electrode and method thereof |
| CN106520747A (en) * | 2016-09-28 | 2017-03-22 | 广州凯耀资产管理有限公司 | Novel immobilized enzyme preparation, and preparation method and application thereof |
| CN108546331A (en) * | 2018-04-20 | 2018-09-18 | 浙江大学 | The preparation method of enzyme-metal organic frame composite membrane and the application of modified electrode bio-sensing |
| CN108548854A (en) * | 2018-06-22 | 2018-09-18 | 桂林中辉科技发展有限公司 | A kind of sialic acid electrochemistry test paper and its preparation and detection method |
| US20190011426A1 (en) * | 2017-07-07 | 2019-01-10 | Soochow University | Nitrite detection device for detecting nitrite concentration |
| CN109507259A (en) * | 2018-12-25 | 2019-03-22 | 宁波大学 | A kind of working electrode and its enzyme biologic sensor system for detecting hypoxanthine and xanthine |
| CN110023745A (en) * | 2016-12-09 | 2019-07-16 | 东北大学 | The durable biosensor and droplet deposition fixing means based on enzyme |
| CN110320257A (en) * | 2019-07-02 | 2019-10-11 | 吉林省裕林药业有限公司 | A kind of blood glucose sensor and preparation method thereof based on metallic zinc organic transistor |
-
2020
- 2020-12-30 CN CN202011616816.9A patent/CN112798664B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1737560A (en) * | 2005-09-02 | 2006-02-22 | 天津大学 | Static electricity spinning prepared fixed enzymic electrode and method thereof |
| CN106520747A (en) * | 2016-09-28 | 2017-03-22 | 广州凯耀资产管理有限公司 | Novel immobilized enzyme preparation, and preparation method and application thereof |
| CN110023745A (en) * | 2016-12-09 | 2019-07-16 | 东北大学 | The durable biosensor and droplet deposition fixing means based on enzyme |
| US20190011426A1 (en) * | 2017-07-07 | 2019-01-10 | Soochow University | Nitrite detection device for detecting nitrite concentration |
| CN108546331A (en) * | 2018-04-20 | 2018-09-18 | 浙江大学 | The preparation method of enzyme-metal organic frame composite membrane and the application of modified electrode bio-sensing |
| CN108548854A (en) * | 2018-06-22 | 2018-09-18 | 桂林中辉科技发展有限公司 | A kind of sialic acid electrochemistry test paper and its preparation and detection method |
| CN109507259A (en) * | 2018-12-25 | 2019-03-22 | 宁波大学 | A kind of working electrode and its enzyme biologic sensor system for detecting hypoxanthine and xanthine |
| CN110320257A (en) * | 2019-07-02 | 2019-10-11 | 吉林省裕林药业有限公司 | A kind of blood glucose sensor and preparation method thereof based on metallic zinc organic transistor |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112798664B (en) | 2023-03-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Bao et al. | 3D graphene/copper oxide nano-flowers based acetylcholinesterase biosensor for sensitive detection of organophosphate pesticides | |
| Pundir et al. | Biosensing methods for xanthine determination: A review | |
| Lakard et al. | Urea potentiometric biosensor based on modified electrodes with urease immobilized on polyethylenimine films | |
| Palmisano et al. | Simultaneous monitoring of glucose and lactate by an interference and cross-talk free dual electrode amperometric biosensor based on electropolymerized thin films | |
| CN2372689Y (en) | Current biological sensor | |
| Phongphut et al. | A disposable amperometric biosensor based on inkjet-printed Au/PEDOT-PSS nanocomposite for triglyceride determination | |
| TWI450967B (en) | Homogeneously-structured nano-catalyst/enzyme composite electrode, fabricating method and application of the same | |
| Tyagi et al. | Glad assisted synthesis of NiO nanorods for realization of enzymatic reagentless urea biosensor | |
| Shin et al. | A planar amperometric creatinine biosensor employing an insoluble oxidizing agent for removing redox-active interferences | |
| US20080237063A1 (en) | Nanopore structured electrochemical biosensors | |
| Snider et al. | A multiwalled carbon nanotube/dihydropyran composite film electrode for insulin detection in a microphysiometer chamber | |
| Li et al. | Immobilization of acetylcholinesterase on one-dimensional gold nanoparticles for detection of organophosphorous insecticides | |
| Huang et al. | A sensitive H2O2 biosensor based on carbon nanotubes/tetrathiafulvalene and its application in detecting NADH | |
| Wang et al. | Biocomposite of cobalt phthalocyanine and lactate oxidase for lactate biosensing with MnO2 nanoparticles as an eliminator of ascorbic acid interference | |
| CN113813945B (en) | Three-dimensional space network graphene-based polyaniline/PtAg composite catalyst and preparation method thereof | |
| McMahon et al. | Design variations of a polymer–enzyme composite biosensor for glucose: Enhanced analyte sensitivity without increased oxygen dependence | |
| CN108572209A (en) | Electrochemica biological sensor electrode and preparation method thereof, electrochemica biological sensor and its application | |
| CN101315345A (en) | Modified electrode and production method for detecting grape-sugar concentration in non-enzyme condition | |
| Jaffari et al. | Novel hexacyanoferrate (III)-modified carbon electrodes: application in miniaturized biosensors with potential for in vivo glucose sensing | |
| Liu et al. | Enzyme biosensors for point-of-care testing | |
| Osborne et al. | Plastic film carbon electrodes: enzymatic modification for on-line, continuous, and simultaneous measurement of lactate and glucose using microdialysis sampling | |
| Zhao et al. | Unadulterated glucose biosensor based on direct electron transfer of glucose oxidase encapsulated chitosan modified glassy carbon electrode | |
| CN110988080A (en) | Flexible oxygen-enriched bio-enzyme electrode and flexible bio-enzyme sensor based on same | |
| Dai et al. | A highly performing electrochemiluminescent biosensor for glucose based on a polyelectrolyte-chitosan modified electrode | |
| CN110669671B (en) | Continuous monitoring device and method for cell metabolism based on hollow fiber membrane |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |