KR101912344B1 - A high sensitive sensor strip for analyzing glucose and a method of preparing the same - Google Patents

Abstract

The present invention relates to a method for manufacturing a highly sensitive glucose measurement sensor strip and a glucose measurement sensor strip manufactured by the same wherein an ultra-small amount of glucose can be detected. The method for manufacturing a highly sensitive glucose measurement sensor strip of the present invention comprises: a polyaniline low temperature interfacial polymerization step of preparing a polyaniline polymer by polymerizing an aniline polymer by using a low temperature interfacial polymerization method; a de-doping treated polyaniline polymer preparation step of preparing a de-doping treated polyaniline polymer by treating the polyaniline polymer with an alkaline solution; a polyaniline/camphorsulfonic acid complex preparation step of preparing a polyaniline/camphorsulfonic acid complex which is doping-treated by mixing the de-doping treated polyaniline polymer with camphorsulfonic acid; a conductive paste preparing step of preparing polyaniline/camphorsulfonic conductive paste by dispersing the doping-treated polyaniline/camphorsulfonic complex in an organic solvent; a sensor strip preparing step of forming a sensor strip pattern by printing the polyaniline/camphorsulfonic conductive paste on a substrate; and an enzyme attachment step of attaching glucose oxidase enzyme to the sensor strip.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention [0001] The present invention relates to a high-sensitivity glucose measurement sensor strip,

The present invention relates to a sensor strip and a method of manufacturing the same, and more particularly, to a method of manufacturing a high-sensitivity glucose measurement sensor strip and a high-sensitivity glucose measurement sensor strip produced by the manufacturing method.

Approximately 487 million people, or 8.3% of the world 's population, are estimated to be diabetic and approximately 4.9 million patients die each year due to diabetic complications. World diabetes patients are expected to increase by more than 190% in 2050 compared to 2010. Diabetes patients in the Pacific Rim, including Korea, currently have the highest proportion of diabetics when they divide the world into 7 regions with 137 million people. In fact, diabetic patients can not easily cope with exercise or dietary control alone. It is a chronic disease that can not be cured by any means once it occurs. Therefore, it is essential for the patients to know the self-test and to manage their lives for the improvement of diabetes.

In general, the self-monitoring blood glucose test is carried out through an electrochemical analysis through a test strip, that is, a sensor strip, a sensor strip is inserted into an inlet of the blood glucose meter, and a small amount of blood is drawn into a sensor strip The blood is automatically sucked in and the result is displayed on the screen of the blood glucose meter.

The sensor strip includes an electrode made of a material capable of conducting electricity to induce an electrochemical reaction with blood on the insulator of a certain size by vapor deposition or the like.

However, the frequent blood collection procedure for blood glucose measurement involves a painful process of stabbing six fingertips a day, and this blood glucose measurement is particularly painful and inconvenient in patients with problems such as endophthacia.

In this regard, Korean Patent Laid-Open No. 10-2011-0073993 has proposed a noninvasive self-glucose meter using a blood glucose near infrared ray (NIR) negative pressure, but has a limitation in that it can not provide a practical solution.

As another prior art, Patent No. 10-0775669 discloses a portable bloodless blood sugar measuring device for cuff based on analysis of blood glucose near infrared ray (NIR) absorbance spectrum, but it makes measurement accuracy compared with blood sampling method Many diabetics still suffer because they can not.

However, since the blood glucose sensor according to the related art including the prior art is basically manufactured to detect glucose contained in the blood, the concentration of glucose that can basically react is high, and it is very difficult to react with the trace amount of glucose contained in the saliva Has a disadvantage. Therefore, there is a need to develop a biosensor having high sensitivity that can detect low concentrations of glucose contained in saliva.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made to solve the above-mentioned problems, and it is an object of the present invention to provide a biosensor for detecting glucose in a trace amount of glucose. However, these problems are illustrative, and thus the scope of the present invention is not limited thereto.

According to one aspect of the present invention, there is provided a method for preparing a polyaniline polymer by polymerizing an aniline monomer using a low temperature interface polymerization method, Treating the polyaniline polymer with a basic solution to prepare a doped polyaniline polymer; Preparing a polyaniline / camphorsulfonic acid complex by mixing the doped polyaniline polymer with camphorsulfonic acid to prepare a doped polyaniline / camphorsulfonic acid complex; A conductive paste preparation step of dispersing the doped polyaniline / camphorsulfonic acid complex in an organic solvent to produce a polyaniline / camphorsulfonic acid conductive paste; A sensor strip manufacturing step of printing the polyaniline / camphorsulfonic acid conductive paste on a substrate to form a sensor strip pattern; And an enzyme attachment step of attaching a glucosoxidase enzyme to the sensor strip.

According to another aspect of the present invention, there is provided a high-sensitivity glucose measurement sensor strip produced by the above-described manufacturing method.

According to another aspect of the present invention, there is provided a pattern formed by printing a conductive polyaniline polymer / camphorsulfonic acid composite paste prepared by dispersing a composite of a polyaniline polymer / camphorsulfonic acid produced by a low temperature interfacial polymerization in an organic solvent A highly sensitive glucose measurement sensor strip is provided, coupled with glucosoxidase.

The glucose sensor was immobilized on the surface of the polyaniline / camphorsulfonic acid conductive paste as the sensor material of the present invention to produce a sensor for detecting blood sugar through saliva having high sensitivity. It was confirmed that a very small amount of glucose in the saliva was detected.

In particular, since the sensor strips through the blood glucose have high levels of glucose in the blood, it is not necessary to detect a trace amount of glucose. However, in the case of detection through saliva, a trace amount of glucose is required to be detected by about 1/20 Instead of the conventional method of making this detection possible by using material and having to take blood every time with pain, anyone can easily detect the glucose level in the blood by means of saliva without pain.

Since the sensor strip of the present invention is manufactured through the screen printing method, it is possible to pattern the polyaniline material into the sensor strip without any difficulty, and the sensor strip can be manufactured quickly without spreading the thickness of the pattern and the distance between the electrodes.

In addition, since the present invention is simply manufactured by the screen printing method, it is advantageous in terms of cost efficiency.

FIG. 1 is a photograph of a doped polyaniline / camphorsulfonic acid composite prepared according to an embodiment of the present invention by a scanning electron microscope (SEM).
FIG. 2 is a photograph of a polyaniline / camphorsulfonic acid conductive paste prepared according to an embodiment of the present invention by a scanning electron microscope (SEM).
3 is a schematic view illustrating a process of fabricating a sensor strip through a screen printing method using a polyaniline / camphorsulfonic acid conductive paste prepared according to an embodiment of the present invention.
FIG. 4 is a schematic diagram (left side) of a sensor strip pattern for screen printing made according to an embodiment of the present invention, and actually photographs (middle) of a sensor strip It is a photographed picture (right).
FIG. 5 is a graph showing a current change rate when glucose is injected into a glucose sensor strip prepared according to an embodiment of the present invention.
FIG. 6 is a graph showing a current change rate when saliva containing various concentrations of glucose is injected into a glucose sensor strip prepared according to an embodiment of the present invention.
FIG. 7 is a graph showing the relationship between non-target substances (PBS, uric acid and ascrobic acid) and target substances (glucose and glucose) in order to confirm the selectivity of the glucose measurement sensor strip prepared according to an embodiment of the present invention. Glucose-containing saliva) is injected into a blood vessel
FIG. 8 is a graph showing a correlation between the rate of change of current measured using a glucose sensor strip prepared according to an embodiment of the present invention and the concentration of glucose in saliva. FIG.

According to one aspect of the present invention, there is provided a method for preparing a polyaniline polymer by polymerizing an aniline monomer using a low temperature interface polymerization method, Treating the polyaniline polymer with a basic solution to prepare a doped polyaniline polymer; And a polyaniline / camphorsulfonic acid complex preparation step of preparing a doped polyaniline / camphorsulfonic acid complex by mixing the doped polyaniline polymer with camphorsulfonic acid; And a conductive paste preparation step of dispersing the doped polyaniline / camphorsulfonic acid complex in an organic solvent to produce a polyaniline / camphorsulfonic acid conductive paste; And a sensor strip manufacturing step of printing the polyaniline / camphorsulfonic acid conductive paste on a substrate to form a sensor strip pattern; And an enzyme attachment step of attaching a glucosoxidase enzyme to the sensor strip.

The principle of measurement of salivary glucose by the sensor strip for measuring high sensitivity glucose prepared by the method according to an embodiment of the present invention is as follows:

When glucose in the saliva contacts with the sensor strip, the glucose is oxidized through the glucosoxidase enzyme on the surface, and the redox couple inside the glucose oxidase is reduced, which is again oxidized by oxygen to produce hydrogen peroxide. At this time, due to the high conductivity of the polyaniline / camphorsulfonic acid conductive paste, it is possible to measure a small amount of the oxidation current change flowing to the surface. Since the current change rate depends on the glucose concentration, It is possible to accurately measure the concentration of glucose in an unknown sample by measuring the current change rate of an unknown sample.

Unless otherwise specified herein, numerical ranges such as temperature, content, size and the like refer to ranges within which the manufacturing method of the present invention can be optimized.

In the above method, the polyaniline low-temperature interfacial polymerization may be performed by forming an interface with an acid aqueous solution and a water-insoluble organic solvent, and then mixing the aniline monomer and the initiator in an interfacial formation solvent and reacting at low temperature.

In the above production method, the acid aqueous solution may be prepared by dissolving one or more acids selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid, and acetic acid in distilled water.

In the above production method, the water-insoluble organic solvent may be one or a mixture of two or more selected from the group consisting of toluene, chloroform, m-cresol, benzene, and n-hexane.

In the above production method, the weight ratio of the aqueous acid solution and the water-insoluble organic solvent may be 1: 1 to 1: 3.

In the above production process, the aniline monomer may be added in an amount of 0.01 to 10 wt% based on the aqueous acid solution. When the amount of the aniline monomer is lower than 0.01 wt% based on the aqueous solution of the acid, the amount of the aniline monomer to be polymerized is small and the yield is low. When the aniline monomer is higher than 10 wt% based on the aqueous acid solution, the aniline is not sufficiently dispersed in the solvent, .

In the above production method, the initiator may be one or two or more selected from the group consisting of ammonium persulfate (APS), divalent copper chloride (CuCl ₂ ), and trivalent iron chloride (FeCl ₃ ). Ammonium persulfate is most preferable, but not limited thereto, and the initiator may be added in an amount of 0.5 to 3 parts by weight based on the aniline monomer. If the amount of the initiator is less than 0.5 times the mass of the aniline monomer, the polymerization may not proceed in all the monomers. If the amount of the initiator is more than 3 times the monomer, the conductivity of the polyaniline polymer may be lowered.

In the above production process, the polymerization temperature in the polyaniline low-temperature interface polymerization step may be -60 to -20 캜, preferably -50 to -30 캜, more preferably -45 to -40 캜 . At this time, the polymerization temperature is not limited to these ranges and may be lower or higher than the above range. However, if the temperature is too low, the polymerization rate becomes very slow. The polyaniline low-temperature interfacial polymerization may be performed by mixing the aniline monomer and the initiator in the interfacial-forming solvent, followed by stirring at a stirring speed of 200 to 400 rpm, and the interfacial low-temperature polymerization may be performed for 10 to 40 hours . If the polymerization time is shorter than 10 hours, the polymerization is not completely completed, and if the polymerization time is longer than 40 hours, the polyaniline polymers may be clumped together and the conductivity may be lowered.

In the above production method, the basic solution may be one in which one or two or more basic substances selected from the group consisting of ammonia, sodium hydroxide, and potassium hydroxide are dissolved in distilled water. Although it is preferable to use an aqueous ammonia solution as the basic solution, it is not limited thereto. The concentration of the basic substance may be 20 to 50 wt%. When the amount of the basic solution is lower than 20 wt%, the doping does not progress smoothly due to the relatively low pH. When the amount of the basic solution is higher than 50 wt%, the conductivity of the polyaniline polymer may be lowered due to an excessively high pH. In addition, the treating time of the basic solution in the step of preparing the depolymerized polyaniline polymer may be 12 to 24 hours. When the doping treatment time is shorter than 12 hours, all of the polyaniline polymer may not be doped, and when it is longer than 24 hours, the conductivity of the polyaniline polymer may be lowered.

In the above production process, in the step of preparing the polyaniline / camphorsulfonic acid complex, the camphorsulfonic acid may be mixed with 0.5 to 3 parts by weight of the polyaniline polymer to be treated. When the amount of camphorsulfonic acid is less than 0.5 times the mass of the polyaniline polymer, the doped polyaniline polymer is not fully doped. When the amount of camphorsulfonic acid is more than 3 times, when the polyaniline / camphorsulfonic acid conductive paste is prepared, It can be lowered. In this case, the mixing time may be 10 to 60 minutes. If the mixing time is shorter than 10 minutes, the doped polyaniline polymer may not be fully doped. If the mixing time is longer than 60 minutes, the conductivity of the polyaniline / camphorsulfonic acid composite may be lowered.

In the above production method, the organic solvent used in the conductive paste production step may be one or a mixture of two or more selected from the group consisting of toluene, chloroform, m-cresol, benzene, and n-hexane. Of these, m-cresol is preferably used, but it is not limited thereto. At this time, the doped polyaniline / camphorsulfonic acid complex may be mixed in the organic solvent in an amount of 1 to 20 wt%. When the amount of the polyaniline / camphorsulfonic acid complex is lower than 1 wt% based on the weight of the entire paste, the viscosity of the solution is too low, so that the printed pattern is spread when the screen printing operation is performed later. When the amount is higher than 20 wt% The screen printing operation may become difficult to proceed.

In the above manufacturing method, in the sensor strip manufacturing step, the printing may be letterpress printing, engraving printing, flat printing, or screen printing, but is more preferably screen printing. In addition, the screen printing may be performed using a screen printing mesh of 100 to 400 mesh. The sensor strip pattern may include a counter electrode, a working electrode, and a reference electrode. In order to make the distance between the counter electrode and the reference electrode constant around the working electrode, As shown in FIG.

The thickness of the sensor strip pattern may be 0.5 to 3 mm, the length may be 20 to 50 mm, and the thickness of the working electrode working area may be set to 2 to 5 mm. However, the size of the sensor strip pattern is not limited to these ranges, and may be smaller or larger than the above range.

In the above production method, the glucosoxidase enzyme may be bound to the working electrode and may be used at a concentration of 20 to 200 mg / ml, preferably at a concentration of 40 to 150 mg / ml, more preferably 50 to 100 mg / 0.0 > mg / ml. < / RTI >

In the implementation of the present invention, the sensor strip for measuring blood glucose through saliva using polyaniline / camphorsulfonic acid conductive paste has a significantly higher sensitivity than the conventional blood glucose sensor strip in detecting saliva glucose, Glucose contained in the saliva can be detected. In addition, the glucose sensor strip using the polyaniline / camphorsulfonic acid conductive paste generally has an advantage that it can reduce the rejection feeling by not using the invasive blood sampling method which is a method of measuring blood glucose every time.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

The embodiments of the present invention are described in order to more fully explain the present invention to those skilled in the art, and the following embodiments may be modified into various other forms, It is not limited to the embodiment. Rather, these embodiments are provided so that this disclosure will be more thorough and complete, and will fully convey the concept of the invention to those skilled in the art. In the drawings, the thickness and size of each layer are exaggerated for convenience and clarity of explanation.

The present inventors have found that a polyaniline polymer is prepared by polymerizing an aniline monomer using a low temperature interfacial polymerization method, then a camphorsulfonic acid is subjected to a doping treatment in a basic solution, and then camphorsulfonic acid is mixed to form a doped polyaniline / camphorsulfonic acid complex (See Fig. 1). FIG. 1 is a photograph of a doped polyaniline / camphorsulfonic acid composite prepared according to an embodiment of the present invention by a scanning electron microscope (SEM). As shown in FIG. 1, a doped polyaniline / camphorsulfonic acid The size of the composite particles was found to be on the order of several tens of nanometers.

Then, the resulting doped polyaniline / camphorsulfonic acid complex was dispersed in an organic solvent to prepare a conductive paste (see FIG. 2). FIG. 2 is a photograph of a polyaniline / camphorsulfonic acid conductive paste prepared according to an embodiment of the present invention by a scanning electron microscope (SEM). As shown in FIG. 2, the doped polyaniline / camphorsulfonic acid composite particles were seamlessly connected with each other, and thus it was confirmed that a paste having good conductivity was produced.

Then, the present inventors manufactured a sensor strip by printing a polyaniline / camphorsulfonic acid conductive paste prepared as described above on a substrate using a screen printing method (see FIG. 3). 3 is a schematic view illustrating a process of fabricating a sensor strip through a screen printing method using a polyaniline / camphorsulfonic acid conductive paste prepared according to an embodiment of the present invention.

FIG. 4 is a schematic diagram (left side) of a sensor strip pattern for screen printing made according to an embodiment of the present invention, and actually photographs (middle) of a sensor strip It is a photographed picture (right). The sensor strip pattern illustrated on the left side of FIG. 4 includes a middle circular working electrode, a counter electrode (relatively long electrode) and a reference electrode (relatively short electrode) on the left and right sides of the working electrode. The electrode pattern of such a sensor strip is illustrative and may be implemented in other shapes. Glucose oxidase enzyme was attached to the working electrode of the generated sensor strip to finally produce a sensor strip for detecting a clorose (the right side of FIG. 4).

After a sample solution containing glucose at a predetermined concentration is dropped using the glucose sensor strip for detecting glucose as described above, a sensor strip having the sample solution dropped thereinto is inserted into a circulating voltage and current meter, (See Fig. 5). FIG. 5 is a graph showing a current change rate when glucose is injected into a glucose sensor strip prepared according to an embodiment of the present invention. As shown in FIG. 5, it was confirmed that the cyclic voltage current was increased according to the concentration of glucose. Furthermore, the sensor strip according to an embodiment of the present invention was able to detect a very low concentration of glucose of 0.18 mg / dL.

Next, in order to confirm whether the glucose measuring sensor strip can detect and quantify glucose in the saliva, the present inventors added glucose to the saliva of the normal person and the saliva of the normal person at a predetermined concentration, (Fig. 6). FIG. 6 is a graph showing a current change rate when saliva containing various concentrations of glucose is injected into a glucose sensor strip prepared according to an embodiment of the present invention. As shown in FIG. 6, the glucose sensor strip according to an embodiment of the present invention can detect glucose in saliva very accurately. In particular, the content of glucose in the saliva of a normal person was lower than the lowest glucose concentration of 0.18 mg / dL measured in FIG. 5, but the sensor strip according to an embodiment of the present invention can discriminate the difference. Therefore, it can be seen that the glucose sensor strip according to an embodiment of the present invention has a very high sensitivity.

In order to confirm whether the glucose sensor strip according to an embodiment of the present invention reacts specifically with glucose only, the inventors of the present invention conducted experiments on a sensor strip in which PBS, uric acid, and ascorbic acid (vitamic C) And the rate of current change was measured by inserting it into a voltage measuring device (Fig. 7). FIG. 7 is a graph showing the relationship between non-target substances (PBS, uric acid and ascrobic acid) and target substances (glucose and glucose) in order to confirm the selectivity of the glucose measurement sensor strip prepared according to an embodiment of the present invention. Glucose-containing saliva) is injected into the blood. As a result, as shown in FIG. 7, it was found that the glucose measurement sensor strip according to an embodiment of the present invention was not reacted with substances such as PBS, uric acid solution and ascorbic acid. This shows that the glucose measurement sensor strip according to an embodiment of the present invention is glucose-specific.

Finally, the inventors of the present invention prepared a calibration curve (see FIG. 8) in order to confirm whether the glucose measurement method using the glucose measurement sensor strip of the present invention can accurately quantify glucose concentration in saliva. FIG. 8 is a graph showing a correlation between the rate of change of current measured using a glucose sensor strip prepared according to an embodiment of the present invention and the concentration of glucose in saliva. FIG. As shown in FIG. 8, it was confirmed that a parabolic curve was drawn between the content of glucose in the saliva and the current change rate. Therefore, the accurate saliva glucose content corresponding to the current change rate measured using the parabolic equation can be quantified.

Hereinafter, the present invention will be described in detail with reference to examples. The present invention may, however, be embodied in many different forms and should not be construed as being limited to the exemplary embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the concept of the invention to those skilled in the art. It is provided to fully inform you.

Example  One: Polyaniline  Polymer polymerization

In order to polymerize polyaniline polymer using low temperature interfacial polymerization method, the interface was made with 40 g of 25 wt% aqueous hydrochloric acid solution and 80 g of chloroform, 2 g of aniline monomer and 2 g of ammonium persulfate were added, Followed by stirring at a temperature for 24 hours to polymerize the aniline monomer to obtain about 0.8 g of a polyaniline polymer.

Example  2: Polyaniline  Polymer polymerization

The polymerization reaction was carried out in the same manner as in Example 1 except that the amount of ammonium persulfate was adjusted to 3 g. As a result, the same results as in Example 1 were obtained.

Example  3: Polyaniline  Polymer polymerization

Experiments were performed in the same manner as in Example 1 except that the amount of ammonium persulfate was adjusted to 4 g to perform the polymerization reaction. As a result, the same results as in Example 1 were obtained.

Example  4: Polyaniline  Polymer polymerization

The polymerization reaction was carried out in the same manner as in Example 1 except that the stirring temperature was adjusted to -50 ° C. As a result, the same results as in Example 1 were obtained.

Example  5: Polyaniline  Polymer polymerization

The polymerization reaction was carried out in the same manner as in Example 1 except that the reaction time was adjusted to 18 hours. As a result, the same results as in Example 1 were obtained.

Example  6: Polyaniline  Polymer polymerization

The polymerization was carried out in the same manner as in Example 1 except that the reaction time was adjusted to 30 hours. As a result, the same results as in Example 1 were obtained.

Example  7: Deodorizing Polyaniline  Polymer manufacturing

The polyaniline polymer prepared in Example 1 was immersed in a 30 wt% ammonia aqueous solution for 24 hours to prepare a doped polyaniline polymer. The above-mentioned dedoping is a step of removing the aniline monomer produced in the polymerization process of aniline by the hydrochloric acid through the aqueous ammonia solution. In the chemical structure, the polyaniline salt (emeraldine) is oxidized to become a polyaniline base (emeraldine).

Example  8: Deodorizing Polyaniline  Polymer manufacturing

The experiment was carried out in the same manner as in Example 7, except that the concentration of the ammonia aqueous solution was adjusted to 20 wt% to perform the doping. As a result, the same result as in Example 7 was obtained.

Example  9: Deodorizing Polyaniline  Polymer manufacturing

The experiment was conducted in the same manner as in Example 7 except that the concentration of the aqueous ammonia solution was adjusted to 40 wt% to perform the doping. As a result, the same result as in Example 7 was obtained.

Example  10: Camphorsulfonic acid  Doped Polyaniline / Camphorsulfonic acid  Composite manufacturing

1 g of the doped polyaniline polymer prepared in Example 7 was mixed with the same amount of camphorsulfonic acid for 20 minutes to prepare a doped polyaniline / camphorsulfonic acid complex.

The polyaniline / camphorsulfonic acid composite thus prepared was photographed using a scanning electron microscope (JSM-6701F, JEOL, USA) (FIG. 1). As a result, as shown in FIG. 1, it was confirmed that the size of the doped polyaniline / camphorsulfonic acid composite particle was about several tens of nanometers.

Example  11: Camphorsulfonic acid  Doped Polyaniline / Camphorsulfonic acid  Composite manufacturing

The procedure of Example 10 was repeated except that the amount of camphorsulfonic acid was 1.5 times the amount of the doped polyaniline polymer to prepare a polyaniline / camphorsulfonic acid complex. As a result, the same result as in Example 10 was obtained.

Example  12: Camphorsulfonic acid  Doped Polyaniline / Camphorsulfonic acid  Composite manufacturing

The procedure of Example 10 was repeated except that the amount of camphorsulfonic acid was twice the amount of the doped polyaniline polymer to prepare a polyaniline / camphorsulfonic acid complex. As a result, the same result as in Example 10 was obtained.

Example  13: Polyaniline / Camphorsulfonic acid  Composite Conductive Paste Production

The polyaniline / camphorsulfonic acid conductive paste was prepared by dispersing the doped polyaniline / camphorsulfonic acid complex prepared in Example 10 at a concentration of 5 wt% in m-cresol.

The polyaniline / camphorsulfone conductive paste prepared above was photographed by scanning electron microscope (FIG. 2). As a result, it was confirmed that the doped polyaniline / camphorsulfonic acid composite particles were seamlessly connected to each other, and thus a paste having excellent conductivity was produced.

Example  14: Polyaniline / Camphorsulfonic acid  Composite Conductive Paste Production

The polyaniline / camphorsulfonic acid complex was prepared by controlling the concentration of the doped polyaniline / camphorsulfonic acid complex dispersed in m-cresol to 1 wt% by the same manner as in Example 13 above. Results The same results as in Example 13 were obtained.

Example  15: Polyaniline / Camphorsulfonic acid  Composite Conductive Paste Production

The polyaniline / camphorsulfonic acid complex was prepared by controlling the concentration of the doped polyaniline / camphorsulfonic acid complex dispersed in m-cresol to 10 wt% by the same procedure as in Example 13 above. As a result, the same result as in Example 13 was obtained.

Example  16: Manufacture of strip pattern frame for screen printing

In order to fabricate a strip pattern frame for screen printing for printing a glucose measuring sensor strip pattern, the distance between the reference electrode and the counter electrode was set to be circular A strip pattern frame for screen printing with a thickness of 2 mm, a length of 30 mm, a working electrode working area of 3 mm and a mesh size of 200 mesh was prepared.

Example  17: Sensor strip pattern printing

The strip pattern frame for screen printing prepared in Example 16 was placed on a substrate, and the polyaniline / camphorsulfonic acid conductive paste prepared in Example 13 was applied and wiped. After stripping the strip pattern frame, A sensor strip printed with a pattern was prepared (Fig. 3).

Fig. 3 is a schematic diagram showing an operation performed in this embodiment. As shown in FIG. 3, it can be confirmed that a pattern is produced by printing a polyaniline / camphorsulfonic acid conductive paste between meshes while performing screen printing.

FIG. 4 is a schematic view (left) of the sensor strip pattern for screen printing prepared in Example 16 and an actual photograph (middle) and an optical microscope photograph (right) of the glucose sensor strip pattern through the saliva prepared in this example, Are shown together. As shown in FIG. 4, it can be seen that a high clarity strip printed with no blurring was produced.

Example  18: Enzyme attached glucose Lateral  Manufacture of sensor strips

Glucose oxidase (Sigma-Aldrich, USA) was dissolved in 0.1 M PBS solution at 10 wt% in the sensor strip prepared in Example 17, and dried for about 6 hours in a nitrogen atmosphere to adsorb polyaniline molecules on the surface of the electrode, thereby preparing a glucose measuring sensor strip.

Example  19: Enzyme attached glucose Lateral  Manufacture of sensor strips

Using the same method as in Example 18, the glucose sensor strip was prepared by adjusting the amount of the glucosoxidase enzyme dilution to 10 μL. As a result, the same results as in Example 18 were obtained.

Example  20: Preparation of enzyme-attached glucose measurement sensor strip

Using the same method as in Example 18, the glucose sensor strip was prepared by adjusting the amount of the glucose oxidase enzyme dilution to 100 μL. As a result, the same results as in Example 17 were obtained.

Experimental Example  1: Measurement of glucose using glucose sensor strip

The artificial saliva mixed with glucose was sequentially diluted with the glucose sensor strip prepared in Example 20, and the strip was inserted into a circulating voltage and current meter (Wonatech, Korea), and a current of 50 mV / sec was supplied , And the rate of current change (? I / I ₀ ) was measured (Fig. 5). As a result, as shown in FIG. 5, it was confirmed that the current change also increases with increasing glucose concentration.

Experimental Example  2: Measurement of glucose in saliva using glucose measurement sensor strip

In order to confirm whether or not the glucose contained in the saliva can be detected, the glucose sensor strip prepared in Example 20 was administered with 0.18 mg / dL, 1.8 mg / dL and 18 mg / dL of saliva was dispensed, and then the rate of current change was measured using a cyclic voltammeter as described above (see Fig. 6). As a result, as shown in FIG. 6, the saliva of the normal person exhibited a lower current change rate than the saliva of 0.18 mg / dL and the glucose content of 0.18 mg / dL, Which is much lower than the detection sensitivity of the glucose meter of the present invention, and that such a trace amount of glucose can be detected by the high sensitivity glucose measurement sensor strip according to an embodiment of the present invention.

Experimental Example  3: False positive test of glucose sensor strip

In order to verify whether a high-sensitivity glucose measurement sensor strip according to an embodiment of the present invention exhibits false-positive by detecting components other than glucose, the inventors of the present invention used 0.1 M phosphate buffer solution (PBS) 0.1 mM uric acid dissolved in phosphate buffer solution, 0.1 mM ascorbic acid dissolved in the phosphate buffer solution and 18 mg / dL of normal saliva and glucose added with 1.8 mg / dL of glucose as a positive control The same experiment as described above was performed on normal human saliva (Fig. 7). As a result, as shown in FIG. 7, the current change was not measured in the other components other than the sugar in the saliva, and it was confirmed that the current change was increased when the concentration was increased by adding glucose to the saliva.

Experimental Example  4: Creating a calibration curve

In order to confirm whether the glucose content in the saliva can be quantified using the glucose measurement sensor strip according to an embodiment of the present invention, the present inventors measured the glucose concentration and current change rate using the correlation between the ΔI / I _O) to prepare a calibration curve (FIG. 8). As a result, it was confirmed that a parabolic curve was drawn between the content of glucose in the saliva and the current change rate as shown in FIG. The accurate saliva glucose content corresponding to the current change rate measured using the parabolic equation can be quantified.

As described above, the highly sensitive glucose detection sensor strip according to an embodiment of the present invention can precisely detect glucose contained in a trace amount in the saliva, and it is possible to accurately detect the glucose content through the correlation between the glucose content and the current change rate It can be precisely quantified.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. I will understand. Accordingly, the true scope of the present invention should be determined by the technical idea of the appended claims.

Claims (27)

An anionic monomer is reacted with an aniline monomer and an initiator by stirring at a temperature of -60 to -30 占 폚 after forming an interface with an acid aqueous solution and a water-insoluble organic solvent, A step of low-temperature interfacial polymerization of polyaniline to produce a polyaniline polymer by polymerization;
Treating the polyaniline polymer with a basic solution to prepare a doped polyaniline polymer;
Preparing a polyaniline / camphorsulfonic acid complex by mixing the doped polyaniline polymer with camphorsulfonic acid to prepare a doped polyaniline / camphorsulfonic acid complex;
A conductive paste preparation step of dispersing the doped polyaniline / camphorsulfonic acid complex in an organic solvent to produce a polyaniline / camphorsulfonic acid conductive paste;
A sensor strip manufacturing step of printing the polyaniline / camphorsulfonic acid conductive paste on a substrate to form a sensor strip pattern; And
And attaching an enzyme to the sensor strip to attach a glucosoxidase enzyme to the sensor strip. delete The method according to claim 1,
Wherein the acid aqueous solution is prepared by dissolving one or more acids selected from the group consisting of hydrochloric acid, sulfuric acid, nitric acid, and acetic acid in distilled water. The method according to claim 1,
Wherein the water-insoluble organic solvent is one or a mixture of two or more selected from the group consisting of toluene, chloroform, m-cresol, benzene, and n-hexane. The method according to claim 1,
Wherein the weight ratio of the acid aqueous solution and the water-insoluble organic solvent is 1: 1 to 1: 3. The method according to claim 1,
Wherein the aniline monomer is added in an amount of 0.01 to 10 wt% based on the aqueous acid solution. The method according to claim 1,
Wherein the initiator is one or more selected from the group consisting of ammonium persulfate (APS), divalent copper (CuCl ₂ ), and trivalent iron (FeCl ₃ ). The method according to claim 1,
Wherein the initiator is added in an amount of 0.5 to 3 parts by weight based on the aniline monomer. The method according to claim 1,
And the polymerization temperature in the polyaniline low-temperature interface polymerization step is from -45 to -40 ° C. The method according to claim 1,
Wherein the polyaniline low-temperature interface polymerization step is carried out by mixing the aniline monomer and the initiator in the interface forming solvent, followed by stirring at a stirring speed of 200 to 400 rpm. The method according to claim 1,
Wherein the polyaniline low temperature interface polymerization step is performed for 10 to 40 hours. The method according to claim 1,
Wherein the basic solution is one in which at least one basic substance selected from the group consisting of ammonia, sodium hydroxide, and potassium hydroxide is dissolved in distilled water. The method according to claim 1,
Wherein the concentration of the basic substance in the basic solution is 20 to 50 wt%. The method according to claim 1,
Wherein the basic aqueous solution treatment time in the step of preparing the depolymerized polyanilyl polymer is 12 to 24 hours. The method according to claim 1,
In the step of preparing the polyaniline / camphorsulfonic acid composite, the camphorsulfonic acid is mixed with 0.5 to 3 parts by weight of the polyaniline polymer subjected to the doping treatment. The method according to claim 1,
Wherein the mixing time of the doped polyaniline polymer and the camphorsulfonic acid in the step of preparing the polyaniline / camphorsulfonic acid complex is 10 to 60 minutes. The method according to claim 1,
Wherein the organic solvent is one or a mixture of two or more selected from the group consisting of toluene, chloroform, m-cresol, benzene, and n-hexane. The method according to claim 1,
In the conductive paste preparation step, the doped polyaniline / camphorsulfonic acid complex is mixed in an organic solvent in an amount of 1 to 20 wt%. The method according to claim 1,
Wherein the printing is letterpress printing, engraving printing, flat printing, or screen printing. 20. The method of claim 19,
Wherein the printing is screen printing. 21. The method of claim 20,
Wherein the screen printing is performed using a screen printing mesh of 100 to 400 mesh. The method according to claim 1,
Wherein the sensor strip pattern comprises a counter electrode, a working electrode, and a reference electrode. 23. The method of claim 22,
Wherein the sensor strip pattern is formed in a circular or semicircular shape so that distances between the reference electrode and the counter electrode are constant with respect to the working electrode. 23. The method of claim 22,
Wherein the glucosoxidase enzyme is bound to the working electrode. The method according to claim 1,
Wherein the glucosoxidase enzyme is added at a concentration of 20 to 200 mg / ml. A sensor strip for measuring a high-sensitivity glucose, which is produced by the method of any one of claims 1 to 24. An acid aqueous solution and a water-insoluble organic solvent, followed by mixing an aniline monomer and an initiator in an interfacial formation solvent and stirring at a temperature of -60 to -30 占 폚. The polyaniline polymer / Sensitive glucose measurement sensor strip having a glucose oxidase bound on a pattern formed by printing a conductive polyaniline polymer / camphorsulfonic acid composite paste in which a complex of camphorsulfonic acid is dispersed in an organic solvent.

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