CN113820375A - Electrochemical sensor for measuring selected characteristic substances in body fluid and preparation method - Google Patents
Electrochemical sensor for measuring selected characteristic substances in body fluid and preparation method Download PDFInfo
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- G—PHYSICS
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- 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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
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Abstract
The invention provides an electrochemical sensor for measuring selected characteristic substances in body fluid and a preparation method thereof.
Description
Technical Field
The invention relates to the technical field of electrochemical biosensors, in particular to an electrochemical sensor for measuring selected characteristic substances in body fluid and a preparation method thereof.
Background
In recent years, the change of certain life styles causes the rapid increase of patients with chronic diseases, in order to strengthen the monitoring management of the disease conditions, the requirements of the patients on a household monitoring system, namely instruments and an electrochemical sensor are improved year by year, the patients can know the self physical conditions in time by strengthening the monitoring of indexes such as blood sugar, uric acid, blood ketone and the like, and the indexes are used as references to adjust the self dietary structure or facilitate doctors to further monitor data to adjust the dosage of the medicine. Therefore, the accuracy and precision of such disposable sensors are important in the daily monitoring of diseases, and they must be easy to mass produce and inexpensive. Most electrochemical sensors in the market are produced by a screen printing process, namely, an electrode system is formed on a substrate made of materials such as polyethylene terephthalate (PET), polyvinyl chloride (PVC), Polycarbonate (PC) and the like by screen printing of conductive materials such as carbon, silver, gold and the like, a certain amount of reaction reagents are dripped on a reaction area of an electrode, a detection channel is formed above the reaction area by a middle partition layer and a covering layer after the reagents are formed into a film, and finally a product is formed. During detection, the body fluid reacts with the reaction reagent in the channel to generate a detectable signal, and the instrument obtains a detection result according to the signal.
The quantity of the reaction reagent participating in the reaction in the detection channel and the thickness of the membrane on the electrode can be influenced by negative and uniform film formation of the reaction reagent, so that the redissolution speed and the reaction rate of the reagent after body fluid sample introduction are influenced, and the accuracy and precision of the sensor are further influenced. In the production process, the phenomenon that a reaction reagent in a reaction area is not full or overflows is found, after the reaction reagent is formed into a film, the problem of uneven thickness of the film is caused under the condition that the reaction reagent is not full, the problem of insufficient amount of the reaction reagent and/or uneven thickness of the film is caused under the condition that the reaction reagent overflows, and if the problems can be solved, the testing accuracy and precision of the electrochemical sensor are further improved.
Disclosure of Invention
In view of the above situation, the present invention aims to provide an electrochemical sensor for measuring a selected feature substance in a body fluid and a method for preparing the same, which can solve the problems of insufficient reaction reagent amount and uneven film thickness caused by the underfill or overflow of a reaction reagent, and further, the accuracy and precision of the measurement.
The invention provides an electrochemical sensor for measuring a selected characteristic substance in body fluid, which comprises a substrate, an electrode system and a protective layer, wherein a reaction reagent capable of reacting with the characteristic substance is arranged on the electrode system, the electrochemical sensor is characterized in that the protective layer is partially deleted to form a deletion part, a hoop consisting of at least one hoop reinforcement is arranged in the deletion part, the hoop comprises at least one interface and a cavity, the reaction reagent is uniformly formed into a film in the cavity by the action force of the interface on the edge of the reaction reagent, and the action force is pulling force and/or pushing force.
Still further, the cuff is constructed of a hydrophilic material.
Furthermore, the number of the stirrups is N, and N is more than or equal to 1.
Further, the ferrule includes a first interface and a second interface; further, the first interface is formed of a hydrophilic material and the second interface is formed of a hydrophobic material.
Furthermore, a hydrophobic part is arranged between the hoop and the edge of the protective layer missing part.
Furthermore, the thickness of the interface is 0.1-1 mm, and the height of the interface is 40-100 um.
Further, the height of the first interface is less than the height of the second interface.
The invention provides a preparation method of an electrochemical sensor for measuring selected characteristic substances in body fluid, which comprises the following steps:
providing a substrate;
providing an electrode system on a substrate;
providing a protective layer having a missing portion on the electrode system;
a collar is disposed over the electrode system, the collar being visible from the absence, and a reagent disposed in a cavity of the collar.
Still further, the method includes the step of providing a hydrophobic portion between the missing portion edge and the cuff.
Furthermore, the hoop is arranged in any one of printing, spraying, coating, sputtering and the like, or any two or more of the above manners.
Furthermore, after the hoop is arranged, the first interface is sprayed with hydrophilic materials, and the second interface is sprayed with hydrophobic materials.
The invention provides an application of an electrochemical sensor for measuring selected characteristic substances in body fluid, which comprises the following steps:
providing a body fluid containing said selected feature substance;
providing an electrochemical sensor prepared by the preparation method of the electrochemical sensor for measuring the selected characteristic substances in the body fluid or the electrochemical sensor for measuring the selected characteristic substances in the body fluid;
the body fluid is mixed with a membrane-forming reaction reagent in a cavity of the collar on the electrochemical sensor, and the selected feature substance reacts with the reaction reagent; the electrochemical sensor outputs a signal of the selected feature substance.
Furthermore, the method also comprises the step of enabling liquid to enter the interior of the sensor through a sample feeding channel formed by arranging a middle separation layer and a covering layer on the substrate of the electrochemical sensor to be mixed with the reaction reagent.
The invention has the beneficial effects that: 1. the problem that the reaction reagent is not full or overflows is solved through the arrangement of the hoop, the reaction reagent is positioned and limited in the cavity, the reaction reagent can be fully and uniformly formed into a film in the cavity, and the accuracy and precision of the test are improved; 2. the arrangement of the hydrophilic first interface and the hydrophobic second interface provides pushing force while providing pulling force for the reaction reagent, so that the reaction reagent can be uniformly formed into a film only in the cavity under the combined action of the two forces; 3. the arrangement of the outer side water-repellent part of the hoop prevents the reaction reagent from contacting the edge of the missing part of the protective layer, meanwhile, reaction reagent thrust is provided, and the phenomenon that quantitative film forming cannot be caused due to overflow of the reaction reagent is prevented.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a schematic structural diagram of two electrochemical sensors for measuring selected characteristic substances in body fluid according to the present invention.
FIG. 2 is a schematic diagram of an electrochemical sensor for measuring a selected feature substance in a body fluid according to one embodiment of the present invention.
Fig. 3 is a schematic structural diagram of an electrochemical sensor provided in the prior art.
Fig. 4 is a schematic view showing the structure of the ferrule of the present invention.
Figure 5 is a schematic diagram of four of the structures of the cuff of the present invention.
FIG. 6 is a schematic diagram of an electrochemical sensor for measuring a selected feature substance in a body fluid according to one embodiment of the present invention.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments, and it should be understood that the described embodiments are some, but not all, embodiments of the present invention.
The present invention provides an electrochemical sensor (hereinafter referred to as electrochemical sensor or sensor) for measuring a selected characteristic substance in a body fluid.
As shown in fig. 1 to 6, the present invention provides an electrochemical sensor, which comprises a substrate 1, an electrode system 2 and a protective layer 3, wherein the substrate 1 is a carrier of the electrochemical sensor, and in the present invention, the substrate is preferably an insulating plastic sheet with certain hardness, including but not limited to PVC, PET, PC; the electrode system 2 is arranged on the substrate 1 and at least comprises a counter electrode and a working electrode, the material of the electrode system 2 can be one or a combination of any conductive substances such as graphite, gold, silver and the like, the electrode system 2 can be obtained by any electrode arrangement mode in the prior art, and particularly, the electrode system can be arranged on the substrate 1 by a screen printing process and other processes; said protective layer 3 is arranged above the electrode system 2, with partial areas of the protective layer 3 missing to form missing portions 31, the protective layer 3 being obtained in any manner known in the art, in particular being able to be built up above the electrode system 2 by means of a screen printing or the like process; the said missing part 31 is set on one end of the protective layer 3, above the electrode system 2, there is a hoop 33, the said hoop 33 can be seen from the missing part 31, the said hoop 33 includes many forms, the hoop 33 includes at least one interface and cavity 32, when setting the reaction reagent to the sensor, the reaction reagent is positioned and limited in the cavity 32, the reaction reagent includes but not limited to glucose reaction reagent, ketone body reaction reagent, lactic acid reaction reagent, uric acid reaction reagent, etc., wherein the said interface is made of hydrophilic material, the edge of the reaction reagent is firmly held by the hydrophilic pull of the interface, thus form the membrane in the cavity 32 evenly; further, the interfaces include a first interface 35 and a second interface 36; in some embodiments, the first interface 35 is made of a hydrophilic material, and the second interface 36 is made of a hydrophobic material, so that the edge of the hydrophilic first interface 35 can firmly grasp the edge of the reactive agent, and the hydrophobic second interface 36 pushes the edge of the reactive agent toward the center of the cavity 32, so that the reactive agent is uniformly stressed under the action of the tensile force and the pushing force and forms a film in the cavity 32, and the thickness of the formed film is more controllable. The first interface 35 and the second interface 36 have a certain thickness, and the thickness is preferably 0.1-1 mm. The heights of the first interface 35 and the second interface 36 can be the same or different, preferably, the height of the second interface 36 is greater than that of the first interface 35, so that the second interface 36 has a stronger thrust effect on the reactant, and the heights of the first interface 35 and the second interface 36 are preferably 40-100 um. The hoop 33 is composed of at least one hoop 34, and the smaller the gap between the hoops 34, the better the effect of the quantitative and stable film formation of the reaction agent, and most preferably the hoop 33 is composed of a single seamless circular hoop 34.
The second interface 36 of the hoop 33 and the edge of the protective layer missing part 31 overlap or separate from each other, as shown in fig. 1 (b), and the edge of the missing part 31 and the second interface 36 of the hoop 33 overlap each other. Preferably, the second interface 36 of the hoop 33 is separated from the edge of the protective layer missing part 31, and a hydrophobic part is arranged between the second interface and the protective layer missing part, and the hydrophobic part provides a pushing force of the reaction reagent to the cavity 32 and a hydrophilic pulling force provided by the interface of the hoop 33 to cooperate with each other to enable the reaction reagent to be quantitatively and uniformly formed into a film in the cavity 32. The arrangement of the hydrophobic part provides the hoop 33 with a pushing force towards the cavity direction, and the reaction reagent is quantitatively and uniformly formed into a film in the cavity by matching with the pulling force action of the hoop 33.
The invention provides that the protective layer 3 is provided with a deletion 31, the deletion 31 being located above the electrode system 2, the collar 33 being located in the position of the deletion 31.
The electrochemical sensor also comprises a middle separation layer 4 and a covering layer 5, wherein the covering layer 5 is arranged above the middle separation layer 4, and the material of the covering layer is preferably hydrophilic material which can accelerate the uniform diffusion of body fluid. The covering layer 5, the middle partition layer 4 and the substrate 1 form a sample feeding channel, and the covering layer 5 is provided with a vent hole 51 at the far end of the sample feeding channel so as to form a siphon effect.
The middle interlayer 4 is arranged above the electrode system 2 and the protective layer 3, the middle part of the middle interlayer 4 is hollowed or forms a notch, the hollowed or notch is positioned above the missing part 31 of the protective layer, and body fluid can enter the electrochemical sensor through a channel formed by the covering layer 5, the middle interlayer 4 and the substrate 1 under the capillary siphoning action of the vent hole 51 to be mixed with a reaction reagent in the cavity 32.
In some embodiments, the body fluid containing the selected characteristic substance may enter the sensor without passing through the sample channel, and may be directly dropped on the reagent to mix with the reagent in a dropping manner, and the selected characteristic substance reacts with the reagent to output a reaction signal, so that the sample channel may not be constructed.
Example 1
This embodiment provides an electrochemical sensor (hereinafter referred to as electrochemical sensor or sensor) for measuring a selected characteristic substance in a body fluid.
As shown in fig. 1 (a), an electrochemical sensor includes a substrate 1, an electrode system 2 and a protective layer 3, wherein a part of the protective layer 3 is hollowed to form a missing part 31, the shape of the missing part 31 is a circle with a seamless circumference, the substrate 1 is a carrier of the electrochemical sensor, and in this embodiment, the substrate is made of PVC material; the electrode system 2 is arranged on the substrate 1 and comprises a counter electrode 21 and a working electrode 22, the electrode system 2 is made of conductive materials, particularly carbon ink, and the electrode system 2 is arranged on the substrate 1 through a screen printing process; the protective layer 3 is arranged above the electrode system 2, and parts of the electrode system 2 are visible from a recess 31 formed by the cutout of the protective layer 3, the recess 31 being located above the electrode system 2. Be equipped with a hoop 33 on electrode system 2 for hold reactant, reactant is glucose reactant, can see hoop 33 from deletion portion 31, hoop 33 includes cavity 32, first interface 35 and second interface 36, the shape of hoop 33 is the closed ring shape, the height at the first interface of hoop 33 and second interface is 50um, thickness between first interface and the second interface is 0.3mm, the first interface of hoop 33 comprises hydrophilic material, and to reactant effect pulling force, the second interface comprises hydrophobic material, and to reactant effect thrust, reactant evenly atress membrane in cavity 32 under the pulling force at first interface and the thrust effect at second interface forms the reaction membrane of thickness homogeneity. The second interface of the band 33 does not overlap with the edge of the missing part 31 or may overlap with the edge of the missing part 31, and when the second interface of the band 33 does not overlap with the edge of the missing part 31, the gap between the second interface of the band 33 and the edge of the missing part 31 is a blank substrate or a hydrophobic part provided with hydrophobic properties.
The covering layer 5 provided by the embodiment is arranged above the septum layer 4, and the material of the covering layer 5 is a hydrophilic film, so that the uniform diffusion of body fluid containing selected characteristic substances can be accelerated. The covering layer 5, the middle partition layer 4 and the substrate 1 form a sample feeding channel, and the covering layer 5 is provided with a vent hole 51 at the far end of the sample feeding channel so as to form a siphon effect.
The middle partition layer 4 provided by the embodiment is constructed above the electrode system 2 and the protective layer 3, a gap is formed in the middle of the middle partition layer 4, the part of the electrode system 2 is exposed, and the body fluid of the selected characteristic substance can automatically enter the sample under the capillary siphonic action of the vent hole 51 through a channel formed by the covering layer 5, the middle partition layer 4 and the substrate 1.
Example 2
This example provides an electrochemical sensor for measuring a selected feature substance in a body fluid, which is the same as example 1 except that the first and second interfaces of the cuff 33 are rectangular as shown in fig. 2.
Example 3
This embodiment provides an electrochemical sensor for measuring a substance with a selected characteristic in a body fluid, as shown in fig. 4 and 5 (b), compared with embodiment 1, the electrochemical sensor is distinguished in that a protective layer 3 is partially missing to form a missing portion 31, the missing portion 31 extends to an end edge to form a notch, a hoop 33 is disposed in the missing portion 31, a hydrophobic portion is disposed between the edge of the missing portion 31 and the hoop 33, the hoop 33 is composed of a hoop 34 with an opening at one end, the hoop 33 has a first interface and a second interface, the first interface is composed of a hydrophilic material and has a height of 50um, the second interface is composed of a hydrophobic material and has a height of 70um, the thickness between the first interface and the second interface is 0.2mm, a reaction reagent is uniformly forced into a membrane in a cavity 32 under the action of a hydrophilic pulling force of the first interface of the hoop 33 and a hydrophobic pushing force of the second interface and the hydrophobic portion, a reaction film having a uniform thickness is formed.
Example 4
This example provides an electrochemical sensor for measuring a selected characteristic substance in a body fluid, as shown in fig. 5 (c), which is different from example 1 in that the cuff 33 includes 4 beads 34; as shown in FIG. 5 (d), the hoop 33 includes n hoops 34, where n ≧ 1. The height of the first interface and the second interface of the hoop 33 is not limited, preferably 40-100 um, the height of the second interface is equal to the first interface, or less than the first interface, or more than the first interface, the thickness between the first interface and the second interface is not limited, preferably 0.1-1 mm.
Example 5
This example provides an electrochemical sensor for measuring a selected characteristic substance in a body fluid, which is different from example 1 in that the first interface and the second interface of the cuff 33 are both made of a hydrophilic material, and the second interface of the missing part 31 and the second interface of the cuff 33 do not overlap. Further, a hydrophobic portion is disposed between the edge of the missing portion 31 and the second interface of the ferrule 33.
Comparative example
This comparative example provides an electrochemical sensor for measuring a selected feature substance in a body fluid, as shown in fig. 2, which is the same as example 1 except that no cuff 33, other arrangements related to the cuff 33, and a hydrophobic portion are provided.
Example 6
The embodiment provides a preparation method of an electrochemical sensor for measuring a selected characteristic substance in body fluid, which comprises the following steps:
the method comprises the steps of providing a substrate 1, arranging an electrode system 2 on the substrate 1, for example, by any one of printing, coating, sputtering, or a combination of two or more of them, and particularly, arranging a conductive material on the substrate 1 in a certain pattern by screen printing.
The protective layer 3 is provided on the electrode system 2, for example by any one of printing, coating, sputtering or a combination of two or more thereof, in particular by screen printing, of an insulating material in a pattern on the electrode system 2.
When the insulating material is provided in a pattern on the electrode system 2 by means of screen printing, the pattern of the hoops 33 may be provided in the missing part 31 of the protective layer 3 and printed together with the protective layer on the electrode system 2.
Alternatively, the band 33 may be separately disposed on the electrode system 2 before or after the protective layer 3 is disposed on the electrode system 2, such that the band 33 is visible from the missing portion 31, for example, by any one of printing, spraying, coating, sputtering, or the like, or any two or more of them in combination.
A reactive agent is disposed in the cavity 32 of the band 33, and a certain amount of the reactive agent is disposed in the cavity 32 of the band 33 in a dropwise manner or in a coating manner to form a film.
Above the end where the collar 33 is located, a septum layer and a covering layer are built up, which is located above the septum layer, so that the base, the septum layer and the covering layer together form the sample inlet channel.
In some embodiments, the sample introduction channel does not need to be formed, and the middle separation layer and the covering layer do not need to be arranged.
In some embodiments, a hydrophobic portion is provided between the edge of the missing portion 31 and the cuff 33.
In some embodiments, after the band 33 is disposed on the electrode system 2, a material of the same or different properties is sprayed or coated onto the first interface and the second interface of the band 33, respectively, e.g., spraying a hydrophilic material onto the first interface and spraying a hydrophobic material onto the second interface.
Example 7
The present embodiment provides a use of an electrochemical sensor for measuring a selected feature substance in a body fluid, comprising the steps of:
preparing an electrochemical sensor for measuring a selected characteristic substance in a body fluid according to any one of examples 1 to 5 or the electrochemical sensor for measuring a selected characteristic substance in a body fluid prepared in example 6;
providing a body fluid containing a selected characteristic substance, and mixing the body fluid with a reaction reagent by directly applying the body fluid in a cavity of a hoop or entering the interior of the sensor through a sample introduction channel formed by a substrate, a middle partition layer and a covering layer to be mixed with the reaction reagent;
the body fluid is mixed with a reaction reagent arranged in the sample introduction channel, the reaction reagent is formed into a film in the cavity of the hoop, the selected characteristic substance in the body fluid reacts with the reaction reagent, the electrochemical sensor outputs a reaction signal of the selected characteristic substance and the reaction reagent to a detection instrument, and the instrument performs analysis and calculation according to the received signal to obtain a test result.
Examples of the experiments
The electrochemical sensors described in the respective examples and comparative examples were used to detect an analyte, which was glucose, as shown in tables 1, 2, 3, 4 and 5. Whether the electrochemical sensor can limit the reaction reagent in the cavity of the hoop 33 can be measured by the abnormal film forming rate of the electrochemical sensor, if the reaction reagent limited in the cavity is abnormal, the testing accuracy of the electrochemical sensor is higher, the abnormal film forming rate in the production process is lower, the overall accuracy of the whole batch of electrochemical sensors is higher, and the glucose concentration refers to the concentration of glucose contained in the prepared blood to be detected; the current is the current value collected from the 5 th second from the beginning of detection, the blood sugar and the current value have a linear relation, the current value at the 5 th second can indirectly reflect the glucose concentration, and the variation coefficient of the current at the 5 th second reflects the precision of the sensor. The second interface of the ferrule 33 of example 1 used in the experiment did not overlap the edge of the absent portion 31 and the void was a blank base.
TABLE 1 reaction reagent film formation abnormality rate
Table 2 example 1 current test results
Table 3 example 2 current test results
Table 4 example 3 current test results
Table 5 comparative example 1 current test results
As seen from the data in Table 1, compared with the comparative example, the film forming abnormal rate of the comparative example is 3.9%, while the film forming abnormal rate of the electrochemical sensor prepared by the preparation method of the electrochemical sensor for measuring the selected characteristic substance in the body fluid is less than 3.9%, so that the detection accuracy of the electrochemical sensor provided by the invention is higher.
As can be seen from the data in tables 2 to 5, compared with the comparative example, in the glucose concentration range of 0 to 605mg/dL, the coefficient of variation of the current at the 5 th second is within 0.4 to 5.2%, and the coefficient of variation of the current at the 5 th second of the comparative example is within 1.9 to 6.9%, and under each glucose concentration, the coefficient of variation of the example is superior to that of the comparative example, so that the precision of the electrochemical sensor for measuring the selected characteristic substance in the body fluid provided by the invention is better, and the precision of the sensor can be improved.
The foregoing detailed description is intended to illustrate and not limit the invention, which is intended to be within the spirit and scope of the appended claims, and any changes and modifications that fall within the true spirit and scope of the invention are intended to be covered by the following claims.
Claims (15)
1. An electrochemical sensor for measuring a selected characteristic substance in body fluid comprises a substrate, an electrode system and a protective layer, wherein a reaction reagent capable of reacting with the characteristic substance is arranged on the electrode system, the electrochemical sensor is characterized in that the protective layer is partially deleted to form a deletion part, a hoop consisting of at least one hoop is arranged in the deletion part, the hoop comprises at least one interface and a cavity, and the reaction reagent is uniformly formed into a film in the cavity by the action force of the interface on the edge of the reaction reagent.
2. The electrochemical sensor of claim 1, wherein the cuff is formed of a hydrophilic material.
3. An electrochemical sensor for the determination of a selected feature in a body fluid according to claim 1 or claim 2, wherein a hydrophobic portion is provided between the cuff and the edge of the absence of the protective layer.
4. An electrochemical sensor for the determination of a selected feature in a body fluid according to claim 1, wherein the applied force is a pulling force and/or a pushing force.
5. The electrochemical sensor of claim 1, wherein the band comprises a first interface and a second interface.
6. The electrochemical sensor according to claim 1 or 5, wherein the thickness of the interface is 0.1-1 mm, and the height of the interface is 40-100 μm.
7. The electrochemical sensor of claim 5, wherein the first interface of the cuff is formed of a hydrophilic material and the second interface is formed of a hydrophobic material.
8. The electrochemical sensor of claim 7, wherein the first interface has a height less than a height of the second interface.
9. The electrochemical sensor of claim 1, wherein the number of stirrups is N, N ≧ 1.
10. A method of making an electrochemical sensor for measuring a selected feature substance in a bodily fluid, comprising the steps of:
providing a substrate;
providing an electrode system on a substrate;
providing a protective layer having a missing portion on the electrode system;
a collar is disposed over the electrode system, the collar being visible from the absence, and a reagent disposed in a cavity of the collar.
11. The method of claim 10, further comprising the step of providing a hydrophobic portion between the edge of the missing portion and the cuff.
12. The method of claim 10, wherein the cuff is applied by printing, spraying, coating, sputtering, or a combination thereof.
13. The method of claim 10, wherein the step of applying the hydrophilic material to the first interface and the step of applying the hydrophobic material to the second interface is performed after the step of applying the cuff is performed.
14. Use of an electrochemical sensor for the determination of a selected feature substance in a body fluid, comprising:
providing a body fluid containing said selected feature substance;
providing an electrochemical sensor according to claims 1-13;
the body fluid is mixed with a membrane-forming reaction reagent in a cavity of the collar on the electrochemical sensor, and the selected feature substance reacts with the reaction reagent;
the electrochemical sensor outputs a signal of the selected feature substance.
15. The use of an electrochemical sensor for the determination of a selected feature in a bodily fluid as claimed in claim 14 wherein the fluid is introduced into the sensor through a sample channel formed by providing a spacer layer and a cover layer over the substrate of the electrochemical sensor to mix with the reagent.
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