CN210990322U - High-sensitivity wearable noninvasive glucose sensor - Google Patents
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- CN210990322U CN210990322U CN201921563067.0U CN201921563067U CN210990322U CN 210990322 U CN210990322 U CN 210990322U CN 201921563067 U CN201921563067 U CN 201921563067U CN 210990322 U CN210990322 U CN 210990322U
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Abstract
The utility model relates to a glucose sensor, especially a wearable noninvasive glucose sensor of high sensitivity, it includes: a base layer (10); a counter electrode layer (20) and a working electrode layer (30) formed on the base layer (10); a nano noble metal particle accumulation layer (40) is formed above the working electrode layer (30); an electron mediator layer (50) is formed on the noble metal nanoparticle deposited layer (40), and a glucose oxidase layer (60) is provided on the electron mediator layer (50). The utility model discloses a design can laminate the stratum basale of skin and make glucose sensor have wearable, through set up nanometer noble metal granule accumulation layer in order to play the catalytic action between glucose oxidase layer and electrode layer to showing sensitivity and the stability that improves glucose sensor, impel the further development and the application of not having wound type glucose sensor.
Description
Technical Field
The utility model relates to a glucose sensor, especially a wearable noninvasive glucose sensor of high sensitivity.
Background
Diabetes mellitus causes the human body to be incapable of maintaining normal blood sugar level, and is one of common diseases which can lead to death and can not be cured. According to the prediction of the international diabetes union, the population of patients with diabetes globally reaches 4.24 hundred million people in 2017, and the population of patients with diabetes reaches 6.28 hundred million people in 2045. Chinese diabetics are more than 1.14 hundred million people, and the number of patients is the first worldwide. Accurate detection of glucose content in human blood is critical to the treatment and prevention of diabetes. Blood glucose monitoring methods can be divided into invasive, minimally invasive and non-invasive measurements. At present, blood sugar is mainly monitored by invasive measurement (such as vein indwelling needle monitoring and fingertip acupuncture monitoring), and the monitoring method not only brings pain to patients, but also is easy to cause wound infection. The minimally invasive measurement is generally to acquire a glucose signal through a probe implanted in the skin of a human body, and although the pain of a patient can be relieved to a certain extent, the minimally invasive measurement needs to be implanted into the skin through an operation, so that the minimally invasive measurement is very inconvenient to use.
Most of the existing noninvasive glucose sensors detect sweat, saliva, tears and other metabolic liquids to determine the blood sugar content of a human body, but the measurement result is inaccurate and the stability is poor due to low glucose content in body fluid and other reasons. However, the tissue fluid has fewer interference factors and has obvious advantages. The tissue fluid is extracted mainly by a reverse ion permeation method, and the working principle is that sodium ions and chloride ions are subjected to electromigration in subcutaneous tissue fluid under the action of an external electric field and respectively move to the positive electrode and the negative electrode of an electrode to form a tiny direct current channel. Because human skin has negative charges, mainly sodium ions migrate to the negative electrode under the action of electric potential to form ion current. The ion flow is used as a channel to carry out neutral glucose molecules in the tissue fluid at the negative electrode of the electrode, and the hyaluronic acid solution is smeared on the surface of the skin to improve the surface charge migration rate so as to achieve the purpose of increasing the output of the tissue fluid. The interstitial fluid can replace blood as a test fluid for detecting blood glucose content. The method can transport molecules through skin without damaging skin surface or contacting blood. However, due to the fact that the amount of extracted human tissue fluid is small, the content of glucose in the extracted human tissue fluid is low, and the like, it is urgently needed to improve the sensitivity and the detection stability of the glucose sensor, and simultaneously improve the biocompatibility of the sensor applied to human skin.
SUMMERY OF THE UTILITY MODEL
Technical problem to be solved
In order to solve the above-mentioned problem of prior art, the utility model provides a wearable noninvasive glucose sensor design of high sensitivity can laminate in the stratum basale of skin through the design and make glucose sensor have wearable, deposits the layer in order to improve catalytic action through set up nanometer noble metal granule between glucose oxidase layer and electrode layer to effectively and improve glucose sensor's sensitivity and stability by a wide margin, impel noninvasive glucose sensor's development and application.
(II) technical scheme
In order to achieve the above object, the utility model discloses a main technical scheme include:
a high-sensitivity wearable noninvasive glucose sensor, comprising:
a base layer (10); a counter electrode layer (20) and a working electrode layer (30) formed on the base layer (10);
a nano noble metal particle accumulation layer (40) is formed above the working electrode layer (30); an electron mediator layer (50) is formed on the nano noble metal particle deposition layer (40), and a glucose oxidase layer (60) is provided on the electron mediator layer (50).
According to a preferred embodiment of the present invention, wherein the working electrode layer (30) and the counter electrode layer (20) are conductive carbon film, metallic silver film, metallic copper film or gold film.
According to a preferred embodiment of the present invention, wherein the working electrode layer (30) and the counter electrode layer (20) are deposited on the substrate layer (10) by evaporation; alternatively, the working electrode layer (30) and the counter electrode layer (20) are formed on the substrate layer (10) by spin coating, brush coating, spray coating, screen printing or printing.
According to a preferred embodiment of the present invention, the nano precious metal particle stacking layer (40) is a stacking layer of nano gold, nano platinum or nano silver particles.
According to a preferred embodiment of the present invention, the nano noble metal particle stacking layer (40) is a stacking layer of nano gold particles.
According to a preferred embodiment of the present invention, the nano noble metal particle deposition layer (40) is formed on the working electrode layer (30) by using a logistic method such as ion beam sputtering, magnetron sputtering or physical vapor deposition.
The ion beam sputtering has the characteristics of simple operation technology, easy and accurate control and suitability for batch production, and can form a deposition layer of nano metal particles with smaller diameters, so that the nano noble metal particles have good catalytic activity, larger specific surface area, roughness and good biocompatibility.
According to a preferred embodiment of the present invention, the nano noble metal particles in the nano noble metal particle stacking layer (40) have a particle size of 0.1nm to 100 nm. Preferably, the nano noble metal particle accumulation layer (40) is a nano gold particle accumulation layer, and the particle diameter of the nano gold particles is less than 5 nm.
According to a preferred embodiment of the present invention, wherein the thickness of the nano noble metal particle stacking layer (40) is 1nm to 1 mm.
According to a preferred embodiment of the present invention, the electron mediator layer (50) is ferric ferrocyanide (PB), ferrocene and its derivatives, Flavin Adenine Dinucleotide (FAD), benzoquinone or polytetrafluoroethylene and organic dyes.
The electron mediator has the advantages of being capable of promoting an electron transfer process, widening a linear range and the like, and a commonly used electron mediator layer (50) is made of PB or FAD. PB can be formed on the nano noble metal particle accumulation layer (40) by adopting an electrochemical deposition method.
According to a preferred embodiment of the present invention, the glucose oxidase layer (60) is formed by fixing glucose oxidase to the electron mediator layer (50) through a porous polymer. The concept of the glucose oxidase layer (60) is known in the art because it is not easy to mold and immobilize a simple glucose oxidase, and in general, it is necessary to perform immobilization and molding (molding into a film or a layer) by dispersing the glucose oxidase in a conductive polymer (preferably, a porous conductive polymer) when manufacturing a glucose sensor.
Preferably, the porous polymer is chitosan or other conductive polymer aerogel. Chitosan is a biocompatible polymer matrix, can well disperse and fix Glucose Oxidase (GOD), has good film forming capability and high water permeability, and also has certain conductivity, so the chitosan is a preferable material for fixing the glucose oxidase.
According to a preferred embodiment of the present invention, wherein the substrate layer (10) is a flexible substrate layer or a hard substrate layer; the flexible base layer is a flexible polymer resin material substrate and the like, and the hard base layer is a hard plastic substrate or hard paper and the like.
According to a preferred embodiment of the present invention, wherein the substrate layer (10) is a flexible substrate layer comprising a lower Polymethylmethacrylate (PMMA) layer (11) and an upper Polyimide (PI) film layer (12), the counter electrode layer (20) and the working electrode layer (30) being arranged above the polyimide film layer (12).
Preferably, the flexible substrate layer has a Young's modulus of 1-200MPa, preferably 4-5MPa, which is good for bending to the skin.
The PMMA layer has high toughness, high stretching rate and high transparency, the high toughness can provide the strength of the whole wearable noninvasive glucose sensor, the high stretching rate can improve the comfort degree of a patient during use, the PMMA layer can be properly bent and stretched according to the body part to be adhered of the user, and the PMMA layer has good fitting property. One surface of the PI film layer faces to the skin side of a user, PI has good thermal stability and good biocompatibility, and can reduce skin discomfort (foreign body sensation) when the PI film layer is tightly attached to the skin. PMMA and PI jointly form a composite flexible substrate, so that the composite flexible substrate has flexibility and tensile rate, can be better attached to skin, improves the comfort level of a user, and improves the biocompatibility of the wearable noninvasive glucose sensor.
(III) advantageous effects
The invention has the beneficial effects that:
the utility model discloses a set up the catalysis layer that one deck nanometer noble metal granule piled up formation at electron media layer and working electrode layer, can promote glucose sensor's detectivity and detection stability by a wide margin, make the application of not having the type glucose sensor of creating of not having type glucose sensor, especially tissue fluid reverse ion penetration method obtain further development.
Experiments prove that the glucose sensor with the nano noble metal particle accumulation layer is 2-3 times that of the glucose sensor without the nano noble metal particle accumulation layer.
The utility model discloses further through with setting up working electrode layer, electron media layer, glucose oxidase layer, counter electrode layer etc. on the compound flexible stratum basale by PMMA and PI, this flexible stratum basale can laminate on user's skin, with the help of the high biocompatibility of PI and PMMA's high toughness and tensile characteristic, improves the biocompatibility of wearable noninvasive glucose sensor to improve user's comfort level.
Drawings
Fig. 1 is a schematic structural diagram of a glucose sensor according to a preferred embodiment of the present invention.
Fig. 2 is a working principle diagram of the glucose sensor of the present invention.
FIG. 3 is an SEM image of the surface of a gold working electrode formed by vapor deposition (ion beam sputtering for 0 second).
Fig. 4 is an SEM image of the surface of the gold nanoparticle stacked layer formed when ion beam sputtering is performed for 60 seconds.
Fig. 5 is an AFM (atomic force microscope) view of a gold film (ion beam sputtering for 0 second) formed on the surface of the working electrode by the vapor deposition method.
Fig. 6 is an AFM image of the surface of the gold nanoparticle stacked layer formed when ion beam sputtering is performed for 60 seconds.
Fig. 7 is a time-current curve of the response of a glucose sensor without a gold nanoparticle-deposited layer prepared when ion beam sputtering was performed for 0 second to glucose.
Fig. 8 is a time-current plot of glucose response for a glucose sensor containing a gold nanoparticle-deposited layer prepared at 60 seconds of ion beam sputtering.
Fig. 9 is a tensile curve of a flexible substrate of the glucose sensor of the present invention.
[ description of reference ]
10 a base layer; 20 pairs of electrode layers; 30 a working electrode layer; a 40 nm noble metal particle build-up layer; 50 an electronic media layer; 60 a glucose oxidase layer; 11 a polymethyl methacrylate layer; 12 polyimide film layer.
Detailed Description
For the purpose of better explaining the present invention and to facilitate understanding, the present invention will be described in detail by way of specific embodiments with reference to the accompanying drawings.
As shown in fig. 1, the wearable noninvasive glucose sensor with high sensitivity of the present invention includes: a substrate layer 10, a counter electrode layer 20, a working electrode layer 30, a nano noble metal particle accumulation layer 40, an electron mediator layer 50 and a glucose oxidase layer 60.
Wherein, a counter electrode layer 20 and a working electrode layer 30 are formed on the substrate layer 10, and a nano noble metal particle accumulation layer 40 is formed above the working electrode layer 30; an electron mediator layer 50 is formed on the noble metal nanoparticle deposited layer 40, and a glucose oxidase layer 60 is provided on the electron mediator layer 50. The characteristics and the functional relationship of the above layers are respectively described as follows:
the working electrode layer 30 and the counter electrode layer 20 are conductive carbon films, metallic silver films, metallic copper films, or gold films, and may be deposited on the substrate layer 10 by evaporation. Preferably, the working electrode layer 30 and the counter electrode layer 20 are gold films formed on the flexible substrate 10 by evaporation, and the thickness is 50-300 nm.
The nano noble metal particle accumulation layer 40 is an accumulation layer of noble metal particles such as nano platinum, nano silver, nano gold and the like, has high conductivity and high catalytic activity, and can effectively enhance the electrocatalytic activity to hydrogen peroxide for a glucose sensor based on hydrogen peroxide detection.
Preferably, the nano noble metal particle stacking layer 40 is a stacking layer of nano gold particles, and the particle diameter of the nano gold particles is 0.1nm-100nm, more preferably ≦ 5 nm. In order to form a layer of deposited gold nanoparticles having catalytic properties, ion beam sputtering is preferably used. The ion beam sputtering method can form not only a stacked layer of the nano gold particles, but also an uneven rough surface (see fig. 4 and 6), thereby obtaining good catalytic activity, a larger specific surface area, a larger roughness and good biocompatibility. Preferably, the thickness of the nano noble metal particle stacking layer 40 is 1nm to 1 mm; the thickness can be precisely controlled by the application time of the ion beam sputtering, which is 5 to 150 seconds.
The electron mediator layer 50 is a reducing compound that promotes the electron transport process and broadens the linear range. The electron mediator layer 50 is made of ferric ferrocyanide (PB Prussian blue), ferrocene and its derivatives, Flavin Adenine Dinucleotide (FAD), benzoquinone, polytetrafluoroethylene, or organic dye. Wherein, when PB is used, it can be deposited on the nano noble metal particle stacking layer 40 by an electrochemical deposition method. The most commonly used material for the electron mediator layer 50 is PB or FAD.
The glucose oxidase layer 60 is formed by fixing glucose oxidase to the electron mediator layer 50 through a porous polymer. Since simple glucose oxidase is not easy to mold and immobilize, in the production of a glucose sensor, it is generally necessary to perform immobilization and molding (molding into a film or a layer) by dispersing glucose oxidase in a conductive polymer (preferably, a porous conductive polymer).
During the preparation of the glucose oxidase layer 60, the glucose oxidase can be dissolved in deionized water, then mixed with the chitosan solution and stirred until the solution is clear and free of bubbles, the mixed glucose oxidase chitosan mixed solution is dripped on the electronic medium layer 50, and the mixed solution is placed at room temperature for 2 to 7 hours until the solvent of the mixed solution is evaporated to dryness. The chitosan is a polymer matrix with biocompatibility, can well disperse and fix the glucose oxidase, has good film forming capability and high water permeability, also has certain conductivity, and is a preferred material for fixing the glucose oxidase.
Further, the substrate layer 10 is a flexible substrate layer, the young's modulus of which is 4-5MPa, and which can be well bent to fit the skin, and it includes a polymethyl methacrylate (PMMA) film layer 11 located below and a Polyimide (PI) film layer 12 located above, a counter electrode layer 20 and a working electrode layer 30 are disposed above the PI film layer 12, and one side of the PI film layer 12 is used to fit the surface of the skin of the person, the PMMA film layer can support and protect the PI film layer, the counter electrode layer 20, the working electrode layer 30, the nano precious metal particle accumulation layer 40, the electronic medium layer 50, and the glucose oxidase layer 60 inside the PI film layer, and the PMMA film layer can be tightly fitted to the skin of the person, and the reaction process as shown in fig. 2 can occur by contacting the cell tissue fluid transmitted. PI has good thermal stability and biocompatibility, and can reduce skin discomfort when closely attached to the skin.
The base layer 10 may be provided as a flexible base layer, and a hard base layer such as a hard plastic sheet substrate or a hard paper substrate may be used. However, in the utility model, the flexible substrate layer of the PMMA + PI composite structure is preferentially adopted, on one hand, the two materials have respectively excellent properties such as stretchability, transparency, high temperature resistance, biocompatibility and the like; on the other hand, the flexible substrate layer of the composite structure has a very suitable young's modulus.
As shown in FIG. 2, glucose is catalyzed by O2Oxidation loses electrons to gluconic acid, electrons and water and O2Generation of H2O2The electrons are transferred by the transfer electron mediator layer 50(PB) to form a current, wherein the nano-gold particles in the nano-noble metal particle accumulation layer 40 play a catalytic role, which can significantly enhance the sensitivity of the sensor. The detection terminal collects current and voltage signals brought by the electron current, and the glucose concentration is quantitatively detected by utilizing the characteristic that the current or voltage signals and the glucose concentration are in a linear relation.
Example one
The high-sensitivity wearable noninvasive glucose sensor of the embodiment can be prepared according to the following method:
firstly, rotating polymethyl methacrylate (PMMA, stirring and dissolving in chlorobenzene at 80 ℃ and 300mg/m L) at 6000 revolutions per minute for 30 seconds to spin-coat on a smooth silicon wafer which is cleaned by UV-ozone treatment and is used for 10 minutes, placing the spin-coated PMMA on a heating plate at 180 ℃ to heat for 10 seconds, then rotating polyamic acid at 6000 revolutions per minute for 30 seconds to spin-coat on a PMMA layer, and preserving heat for one hour at three heating temperatures of 80 ℃, 120 ℃ and 140 ℃ respectively to obtain the flexible substrate 10.
The flexible substrate 10 has a tensile curve, see fig. 9, and a young's modulus of 4.75MPa, which is calculated to be very good at bending, stretching and conforming to the skin.
Step two: the counter electrode layer 20 and the working electrode layer 30 are obtained by depositing gold 50-300nm thick on the prepared flexible substrate 10 by thermal evaporation at an evaporation rate of 0.1-1 angstrom/sec. The counter electrode layer 20 and the working electrode layer 30 were 200nm thick in this embodiment.
Step three: under vacuum conditions, gold nanoparticles are sputtered onto the surface of the working electrode layer 30 (gold film formed by vapor deposition) for 5 to 150 seconds. The grain diameter of the gold nanoparticles formed by sputtering is about 5nm, even is very small (see figures 4 and 6), so that the catalytic activity of the gold nanoparticles is higher. Referring to fig. 3-4, SEM images were detected at 0 seconds and 60 seconds from the start of sputtering. When sputtering is carried out for 60 seconds, the surface of the gold nanoparticles piled up can be observed by SEM.
Referring to fig. 5-6, AFM images measured at 0 seconds and 60 seconds from the start of sputtering are shown. It can be seen that a very uniform and fine (significantly finer than that in fig. 5) deposit of gold nanoparticles with a concave-convex surface is obtained at 60 seconds of sputtering. These nano-gold particle-deposited layers play a good catalytic role and constitute the nano-noble metal particle-deposited layer 40.
Step four: 40.6mg of ferric chloride (FeCl) was taken3) 82.31mg of potassium ferricyanide (K)3Fe(CN)6) And 745.5mg potassium chloride (KCl) in a 100m L volumetric flask, 829 mu L of hydrochloric acid (99% concentration) was added and deionized water was used as a solvent to prepare a ferric ferrocyanide solution, PB was electrochemically deposited on the accumulated layer of gold nanoparticles (i.e., the accumulated layer of noble metal nanoparticles 40) in a three-electrode system at a voltage of 0.4V by chronoamperometry, and then dried at 60 ℃ for 5 minutes.
Step five, dissolving 20mg of chitosan into 2% acetic acid solution, adding 8 mu L glycerol, stirring uniformly at 80 ℃ until the solution is clear and bubble-free, preparing glucose oxidase solution (dissolved in deionized water and 30mg/m L), mixing the chitosan solution and the glucose oxidase solution, stirring until the solution is clear and bubble-free, dripping the mixed glucose oxidase chitosan mixed solution on a PB layer, standing for 2-7 hours at room temperature until the solvent is evaporated to dryness.
Step six: and lifting the completed structure from a silicon chip to obtain the flexible sensor, thus obtaining the high-sensitivity wearable noninvasive glucose sensor with the structure shown in the figure 1.
Fig. 7 is a time-current curve of glucose response of a glucose sensor without a gold nanoparticle stacking layer, and fig. 8 is a time-current curve of glucose response of a glucose sensor with a gold nanoparticle stacking layer prepared by ion beam sputtering for 60 seconds, and responses of two glucose sensors were measured by an electrochemical amperometric method under a three-electrode system. In the case where the thickness and the preparation conditions of the counter electrode layer 20, the working electrode layer 30, the electron mediator layer 50, and the glucose oxidase layer 60 were the same in both glucose sensors, the average current per millimole was 88.11. mu.A/mM in the former, 217.78. mu.A/mM in the latter, and 2.47 times in the latter, as seen from comparison of the two graphs. It can be seen that the response of the glucose sensor with the PB layer modified by the stacking layer of gold nanoparticles is more than twice that of the glucose sensor without the stacking layer of gold nanoparticles.
Further, as a result of comparing and measuring the cyclic voltammograms of the PB layers of the 60-second sputtered gold nanoparticle deposited layer and the 0-second sputtered gold nanoparticle deposited layer, the current peak of the PB layer cyclic voltammogram was more than doubled compared to the current peak of the unputtered (0-second sputtered) gold nanoparticles. Experiments show that the impedance of the PB layer containing the nano-gold particle accumulation layer is reduced, the current generated by oxidation of glucose is easier to detect, and the sensitivity and the detection stability of the glucose sensor are greatly improved.
Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described in the foregoing embodiments, or equivalents may be substituted for elements thereof. Any modification, equivalent replacement, or improvement made within the spirit and scope of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A high-sensitivity wearable noninvasive glucose sensor, characterized in that it comprises:
a base layer (10); a counter electrode layer (20) and a working electrode layer (30) formed on the base layer (10);
a nano noble metal particle accumulation layer (40) is formed above the working electrode layer (30); an electron mediator layer (50) is formed on the nano noble metal particle deposition layer (40), and a glucose oxidase layer (60) is provided on the electron mediator layer (50).
2. The high-sensitivity wearable noninvasive glucose sensor of claim 1, characterized in that the working electrode layer (30) and the counter electrode layer (20) are conductive carbon film, metal silver film, metal copper film or gold film.
3. The high-sensitivity wearable noninvasive glucose sensor of claim 2, characterized in that the working electrode layer (30) and the counter electrode layer (20) are deposited on the substrate layer (10) using evaporation; alternatively, the working electrode layer (30) and the counter electrode layer (20) are formed on the substrate layer (10) by spin coating, brush coating, spray coating, screen printing or printing.
4. High-sensitivity wearable noninvasive glucose sensor according to claim 1 or 2 or 3, characterized in that the nano noble metal particle stack layer (40) is a stack of nano platinum, nano silver or nano gold particles.
5. The high-sensitivity wearable noninvasive glucose sensor of claim 4, characterized in that the nano noble metal particle accumulation layer (40) is formed on the working electrode layer (30) by ion beam sputtering, magnetron sputtering or physical vapor deposition.
6. The high-sensitivity wearable noninvasive glucose sensor of claim 5, characterized in that the nano noble metal particles in the nano noble metal particle accumulation layer (40) have a particle size of 0.1nm-100 nm.
7. High-sensitivity wearable noninvasive glucose sensor according to claim 5 or 6, characterized in that the thickness of the nano noble metal particle accumulation layer (40) is 1nm-1 mm.
8. The high-sensitivity wearable noninvasive glucose sensor of claim 1, characterized in that the substrate layer (10) is a flexible substrate layer or a rigid substrate layer; the flexible base layer is a flexible polymer resin material substrate, and the hard base layer is a hard plastic substrate or hard paper.
9. High-sensitivity wearable noninvasive glucose sensor according to claim 1, characterized in that the substrate layer (10) is a flexible substrate layer comprising a lower polymethylmethacrylate layer (11) and an upper polyimide film layer (12), above which polyimide film layer (12) the counter electrode layer (20) and the working electrode layer (30) are arranged.
10. The high-sensitivity wearable noninvasive glucose sensor of claim 9, wherein the young's modulus of the flexible substrate layer is 1-200 MPa.
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