KR20150006114A - Biochip For Measuring Blood Glucose, and Apparatus For Measuring Blood Glucose For Smart Phone Including The Same - Google Patents

Abstract

Provided are a biochip for measuring blood glucose, and an apparatus for measuring blood glucose for a smart phone including the same. According to an aspect of the present invention, provided is a biochip for measuring blood glucose which includes: a substrate where a blood glucose inflow region for receiving blood glucose and a reaction region connected to the blood glucose inflow region are formed on one side and a recess part is formed in the reaction region; a porous separator which is arranged on the blood glucose inflow region and faces one side of the substrate; and silica nanoparticle and a reaction reagent which are distributed on the reaction region of the substrate. Thereby, blood glucose is easily measured by using a portable biochip for measuring blood glucose and a CMOS image sensor. In particular, blood glucose is easily measured by using a CMOS image sensor installed to a smart phone. And, blood glucose is quantitatively measured and checked by a blood glucose measurement application installed to a smart phone.

Description

[0001] The present invention relates to a biochip for measuring blood glucose and a blood glucose meter for a smartphone including the biochip,

More particularly, the present invention relates to a biochip for blood glucose measurement capable of quantitatively measuring blood glucose according to a change in color intensity according to blood glucose and a reagent, and a blood glucose meter for a smartphone including the biochip .

Recently, there has been a steady increase in the number of patients being treated with diabetes, which is one of the representative adult diseases. Accordingly, interest and demand for the technique for measuring blood glucose of diabetic patients are rapidly increasing.

However, the conventional blood sugar measuring apparatus has a problem that it is troublesome to use an enzyme sensor, for example, every two to three days by using a biological enzyme method. Further, there is a problem in that the structure of the blood glucose measurement device is complicated, making it difficult to manufacture, and bulky and difficult to carry.

Therefore, there is a desperate need for a blood glucose measurement technique capable of improving user convenience based on the real-time blood glucose measurement result ensured with accuracy and reliability in an actual use environment.

Korean Patent Publication No. 10-2007-0006304

An object of the present invention is to provide a biochip for blood glucose measurement that is simple in portability and capable of real-time blood glucose measurement with a simple configuration, and is capable of detecting blood glucose of a blood flowing into a biochip for blood glucose measurement using a CMOS image sensor attached to a conventional smartphone And to provide a blood glucose meter for a smartphone capable of quantitatively measuring blood glucose through a blood glucose measurement application installed in a smartphone.

According to an aspect of the present invention, there is provided a blood pressure monitor comprising: a substrate on which a blood glucose inflow area into which blood glucose flows into a first surface and a reaction area connected with the blood glucose inflow area are formed;

A porous separator disposed on the blood-glucose inflow area opposite to the one surface of the substrate; And

A silica nanoparticle and a reaction reagent distributed on the reaction region of the substrate;

A biochip for measuring blood glucose is provided.

The biochip for blood glucose measurement may further include a cap member on the reaction region.

The cap member includes a convex portion, and the convex portion and the concave portion can mutually combine in a direction in which the convex portion of the convex portion and the concave portion of the concave portion are apart from each other to form a space.

The recess can receive blood glucose and form a photon detection region.

The space can accommodate blood glucose and form a photon detection region.

Wherein the porous separator is selected from the group consisting of nitrocellulose, polysulfone, polyester, polyamide, polyimide, polyamideimide, poly (meta-phenylene isophthalamide), polyetherketone, polyetherimide, For example, made of polypropylene, polypropylene, polyvinylidene fluoride, perfluoropolymers, polyvinyl chloride, polyvinylidene chloride, polyethylene glycol derivatives, polyoxides, polyvinylacetates, polystyrenes, polyacrylonitriles, polymethylmethacrylates and nonwoven fibers And the like.

The silica nanoparticles may be hydrophilic functional group functionalized.

The hydrophilic functional group may be at least one selected from the group consisting of an amine group, a carboxyl group, a hydroxyl group, and a thiol group.

The silica nanoparticles may have a diameter of 10 to 300 nm.

The reaction reagent may include glucose oxidase and chromogen.

The glucose oxidase may include glucose oxidase immobilized on the silica nanoparticles.

The above chromogens are preferably selected from the group consisting of 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid), O- O-Dianisidine, O-Toluidine-Blue, Potassium Iodide, Tetra methyl benzidine, 8-anilino-1-naphthalenesulfonic acid ammonium binding- [3-methyl-2-benzothiazolinone hydrazone] N-sulfonylbenzenesulfonate monosodium combined with 8-anilino-1-naphthalene sulfonic acid (Meta [3-methyl-2-benzothiazolinonehydrazone] N-sulfonyl benzenesulfonate monosodium ammonium, MBTHSB-ANS), 3,3 ', 5,5'-tetramethylbenzidine and syringaldazine, 3,3', 5,5'-tetramethylbenzidine and syringaldazine, Primaquine diphosphate, Thiazole yellow G, and Auramine O-anhydrous.

The reaction reagent may further include peroxidase.

The substrate further forms a blood glucose absorbing region connected to the reaction region in a direction away from the blood glucose inflow region and the biochip for blood glucose measurement further comprises an absorbing member on the blood glucose absorbing region.

The substrate and the cap member may be light transmissive.

Wherein at least one of the substrate and the cap member is formed of a material selected from the group consisting of polyester, polyacrylate, polyethylene (PE), polypropylene (PP), polysulfone (PS), polyethersulfone (PES), polycarbonate , Polyarylate (PAR), polyimide (PI), polyamide, and glass.

Wherein the biochip for blood glucose measurement comprises at least one selected from a first adhesive member between the substrate and the cap member and a second adhesive member between the cap member and the porous separator, A through hole corresponding to the blood glucose inflow area, the reaction area and the blood glucose absorption area is formed, and the blood glucose perforation may be formed corresponding to the blood glucose inflow area so that the second adhesive member passes the blood glucose.

The cap member may be such that one end of the cap member does not overlap with all or a part of the blood glucose penetration hole of the second adhesive member.

The biochip for blood glucose measurement may further include a pad member on the porous separation membrane.

According to another aspect of the present invention,

A substrate on which a blood glucose inflow region into which blood glucose flows into the first surface and a reaction region connected with the blood glucose inflow region are formed, and a recess is formed in the reaction region; A porous separator disposed on the blood-glucose inflow area opposite to the one side of the substrate; And a reaction reagent for reacting with the silica nanoparticles distributed on the reaction region of the substrate. And

A CMOS image sensor for measuring the number of photons transmitted through the biochip for blood glucose measurement;

A blood glucose meter is provided.

The transmitted photon number may be the number of photons transmitted through the recess, which is the photon detection region of the biochip for blood glucose measurement.

The CMOS image sensor may be mounted on a smart phone.

The smartphone may be provided with an application for quantitatively displaying the photon number measured by the CMOS image sensor.

The application may quantitatively display the concentration of blood glucose based on a database measuring the relationship between the concentration of blood sugar and the photon number transmitted through the biochip for measuring blood glucose.

According to the embodiment of the present invention, it is possible to easily perform blood glucose measurement using a biochip for measuring blood sugar and a CMOS image sensor, which are easy to carry, and more particularly, to a blood glucose meter for a smartphone equipped with a CMOS image sensor And blood glucose can be easily measured without regard to time, and blood glucose can be quantitatively confirmed in real time by the installed blood glucose measurement application.

1 is a schematic view of a blood glucose meter according to an embodiment of the present invention.
FIG. 2 is an exploded perspective view of a biochip for measuring blood glucose levels before introduction of silica nanoparticles and a reaction reagent according to an embodiment of the present invention. FIG.
3 is an exploded perspective view of a biochip for measuring blood glucose after introduction of silica nanoparticles and a reaction reagent according to an embodiment of the present invention.
FIG. 4 is a photograph showing the comparison of biochips for measuring blood glucose of the present invention before and after the reaction in blood glucose measurement using four blood samples having different blood glucose concentrations.
5 is a graph showing the relationship between the blood glucose concentration and the photon number transmitted through the biochip for blood glucose measurement according to the present invention.
FIG. 6 is a graph showing the linearity of the blood glucose concentration predicted value and the measured value graph of the biochip for measuring blood glucose of the present invention.
7 is a graph showing the linearity of the blood glucose concentration and the digital measured value graph of the photon number of the blood glucose meter of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS The present invention is capable of various modifications and various embodiments, and specific embodiments are illustrated in the drawings and described in detail in the detailed description. It is to be understood, however, that the invention is not to be limited to the specific embodiments, but includes all modifications, equivalents, and alternatives falling within the spirit and scope of the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

The terms first, second, etc. may be used to describe various components, but the components should not be limited by the terms. The terms are used only for the purpose of distinguishing one component from another.

The terminology used in this application is used only to describe a specific embodiment and is not intended to limit the invention. The singular expressions include plural expressions unless the context clearly dictates otherwise. In the present application, the terms "comprises" or "having" and the like are used to specify that there is a feature, a number, a step, an operation, an element, a component or a combination thereof described in the specification, But do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, or combinations thereof.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. Referring to the accompanying drawings, the same or corresponding components are denoted by the same reference numerals, do.

FIG. 1 is a view schematically showing a blood glucose meter according to an embodiment of the present invention. FIG. 2 is an exploded perspective view of a biochip for blood glucose measurement according to an embodiment of the present invention before introduction of immobilized amine- FIG. 3 is an exploded perspective view of silica nanoparticles of a biochip for blood glucose measurement according to an embodiment of the present invention after introduction of a reaction reagent. FIG.

Referring to FIG. 1, a schematic structure of a blood glucose meter 1 according to an embodiment of the present invention will be described. The blood glucose meter 1 includes a biochip 100 for blood glucose measurement and a CMOS image sensor 200.

By measuring the degree of light transmission to the biochip 100 for blood sugar measurement using the CMOS image sensor 200, particularly the CMOS image sensor 200 mounted on a smartphone (not shown), blood glucose measurement can be performed quickly and effectively have.

2 and 3, the biochip 100 for measuring blood glucose according to an embodiment of the present invention includes a blood glucose inflow region 112 in which blood glucose flows into one side, a reaction region 112 connected to the blood glucose inflow region 112, A substrate 110 on which a concave portion 116 is formed in the reaction region 114; A porous separation membrane (130) disposed on the blood glucose inflow region (112) opposite to the one side of the substrate (110); And a reaction reagent 140 distributed on the reaction region 114 of the substrate 110.

It is preferable that the substrate 110 has a light transmitting property.

As a material of the substrate 110, a material such as polyester, polyacrylate, polyethylene (PE), polypropylene (PP), polysulfone (PS), polyethersulfone (PES), polycarbonate (PC), polyarylate ), Polyimide (PI), polyamide or glass can be used singly or in combination of two or more kinds. Preferably, polyester can be used.

A blood glucose inflow region 112 and a reaction region 114 connected to the blood glucose inflow region 112 are formed on one surface of the substrate 110. The reaction region 114 can be extended in a direction away from the blood glucose inflow region 112 and the blood glucose inflow region 112 and the reaction region 114 are continuously connected and there is no visible boundary. Therefore, they may overlap in some sections.

At this time, a recess 116 can be formed in a part of the reaction region 114, which can hold a certain amount of plasma containing blood sugar. The concave portion 116 can form the photon detection region A together with the convex portion 122 of the cap member 120 to be described below. The photon detection area A will be described in detail below.

When the blood to be measured is supplied to the biochip 100 for blood glucose measurement according to the present embodiment, a plasma component including a blood glucose component excluding a biopolymer substance such as blood cells and proteins is introduced into the blood glucose inflow region (112) and can move to the reaction zone (114). The silica nanoparticles and the reaction reagent 140 are distributed on the reaction region 114. The blood may be whole blood.

The cap member 120 may be further disposed on the reaction region 114 of the substrate 110 and it is preferable that the cap member 120 be made of a light transmissive material similar to the substrate 110. [

Any material that can be used as the material of the substrate 110 as the material of the cap member 120 can be used.

The cap member 120 may form the convex portion 122 at a position corresponding to the concave portion 116 of the substrate 110 and may be formed by a convex portion of the convex portion 122 and a concave portion of the concave portion 116 Can be coupled with each other in the direction in which they are separated from each other to form a space.

The space accommodates a part of the blood plasma containing the blood sugar flowing into the reaction area 114, and the light passing through the space serves as a reference for detecting the photon number by the CMOS image sensor 200. Therefore, the region formed by the concave portion 116 and the convex portion 122 can form the photon detection region A. [

The photon detection area A may be formed only by the recess 116 when the cap member 120 is not used but may preferably be a part where the recess 116 and the protrusion 122 are combined.

Such a photon detection region A can stably accommodate blood plasma containing blood sugar that reacts with a reaction reagent. The thickness of the photon detection region A is greater than the depth of the depression 116 and the depth of the depression 116 122). Therefore, it is advantageous that the light transmittance according to the internal color intensity can be accurately reflected in the sensor. In addition, the reaction time is shortened and an accurate photodetection area is ensured.

The porous separator 130 is disposed at a position corresponding to the blood glucose inflow area 112 on the substrate 110 and filters a biological polymer such as blood cells and proteins from a whole blood including all components of the sample blood. to be.

In some cases, the porous separator 130 may further include a pad 150 to directly receive the sample blood for blood glucose measurement and use it as a primary filtering area. The pad 150 may be made of glass fiber, membrane, or the like, but is not limited thereto. As the membrane, it is possible to use at least one selected from the group consisting of nitrocellulose, polysulfone, polyester, polyamide, polyimide, polyamideimide, poly (meta-phenylene isophthalamide), polyetherketone, polyetherimide, , Polyvinylidene fluoride, perfluoropolymer, polyvinyl chloride, polyvinylidene chloride, polyethylene glycol derivatives, polyoxides, polyvinyl acetate, polystyrene, polyacrylonitrile, or polymethyl methacrylate, And can be used in parallel.

2 and 3, the porous separator 130 disposed on one side of the substrate 110 has one end of the cap member 120 interposed between the porous separator 130 and the substrate 110, And can perform the function of coupling the member 120 to the substrate 110.

The material of the porous separation membrane 130 may be selected from the group consisting of nitrocellulose, polysulfone, polyester, polyamide, polyimide, polyamideimide, poly (meta-phenylene isophthalamide), polyetherketone, Polyvinylidene fluoride, polyvinyl chloride, polyvinylidene chloride, polyethylene glycol derivatives, polyoxides, polyvinyl acetate, polystyrene, polyacrylonitrile, polymethyl methacrylate, Or non-woven fibers can be applied.

It is preferable that the silica nanoparticles and the reaction reagent 140 are disposed on the reaction region 114 of the substrate 110 and the silica nanoparticles are hydrophilic group functionalized.

The hydrophilic functional group is preferably an amine group, a carboxyl group, a hydroxyl group, a thiol group, or the like, more preferably an amine group.

The amine group, carboxyl group, hydroxyl group and thiol group may each independently be C1 to C30, preferably C1 to C20, more preferably C1 to C10, still more preferably C1 to C6.

The amine group may be a primary amine group (-NH ₂ ), or a secondary amine group (-NRH), preferably a primary amine group. Wherein R is an alkyl group.

As used herein, unless otherwise defined, the term "alkyl group" means a straight, branched or cyclic aliphatic hydrocarbon group. The alkyl group may be a "saturated alkyl group" which does not contain any double or triple bonds. The alkyl group may be an "unsaturated alkyl group" comprising at least one double bond or triple bond. The alkyl group may be a C1 to C14 alkyl group. More specifically, it may be a C1 to C10 alkyl group or a C1 to C6 alkyl group.

For example, the C1 to C4 alkyl groups may have 1 to 4 carbon atoms in the alkyl chain, i.e., the alkyl chain may be optionally substituted with one or more substituents selected from the group consisting of methyl, ethyl, propyl, iso-propyl, n-butyl, Indicating that they are selected from the group.

Specific examples of the alkyl group include a methyl group, ethyl group, propyl group, isopropyl group, butyl group, isobutyl group, t-butyl group, pentyl group, hexyl group, ethenyl group, Butyl group, cyclopentyl group, cyclohexyl group, and the like.

The reaction reagent 140 may include glucose oxidase and chromogen. Here, the chromogen serves as a dye exhibiting a predetermined color intensity according to the blood sugar content. Such chromogen may be in a reduced state, which is colorless, and may exhibit a predetermined color intensity depending on the type of chromogen depending on the degree of oxidation. In other words, the higher the degree of oxidation of chromogens, the darker the color.

Any one or more of the reaction reagents and chromogens can be immobilized on silica nanoparticles functionalized with a hydrophilic functional group according to electrostatic attraction.

The silica nanoparticles functionalized with a hydrophilic functional group can help improve the performance of the biochip 100 for measuring blood glucose of the present invention. In general, silica nanoparticles have chemical stability, thermal stability and biological compatibility.

Since the silica nanoparticles are hydrophilic, the plasma containing the blood glucose is uniformly and rapidly allowed to flow in the reaction region where the silica nanoparticles are distributed, so that the blood glucose measurement can be performed accurately and quickly. Preferably, a hydrophilic functional group is bonded to the silica nanoparticle to increase the hydrophilicity, so that the blood glucose can be measured more accurately and more rapidly.

The diameter of the silica nanoparticles is preferably smaller than the wavelength of the visible light. Specifically, it is preferably 10 to 300 nm, more preferably 70 to 200 nm, even more preferably 50 to 100 nm . When the diameter of the silica nanoparticles is smaller than 10 nm, the workability is lowered. When the diameter exceeds 300 nm, the light transmittance is lowered.

The chromogens may be selected from the group consisting of 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid), O- O-Dianisidine, O-Toluidine-Blue, Potassium Iodide, Tetra methyl benzidine, 8-anilino-1-naphthalenesulfonic acid ammonium binding- [3-methyl-2-benzothiazolinone hydrazone] N-sulfonylbenzenesulfonate monosodium combined with 8-anilino-1-naphthalene sulfonic acid (Meta [3-methyl-2-benzothiazolinonehydrazone] N-sulfonyl benzenesulfonate monosodium ammonium, MBTHSB-ANS), 3,3 ', 5,5'-tetramethylbenzidine and syringaldazine, 3,3', 5,5'-tetramethylbenzidine and syringaldazine, Primaquine diphosphate, Thiazole yellow G, Auramine O-anhydrous, and the like.

In the reaction reagent, the content of the glucose oxidase may be 0.01 to 40, more preferably 0.05 to 20, still more preferably 0.1 to 10, in terms of the weight ratio of glucose oxidase to chromogen weight have. Here, if the weight ratio is less than 0.01, the rate of oxidation of blood sugar is decreased. If the weight ratio is more than 40, the rate of oxidation of blood sugar is insignificantly increased.

In addition, the reaction reagent may further include peroxidase. The peroxidase can act as a catalyst promoting the oxidation reaction of the chromogen.

The content of the peroxidase may be 0.01 to 10, preferably 0.05 to 5, more preferably 0.1 to 2, by weight of peroxidase to glucose oxidase. If the weight ratio of peroxidase to glucose oxidase is less than 0.01, the oxidation reaction rate of chromogen is decreased. If the weight ratio is more than 10, the rate of oxidation reaction of chromogen may be insignificant.

Examples of the reaction include the reaction of glucose with oxidase and the reaction of 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) The color intensity change due to the oxidation reaction of ABTS can be represented by the following reaction equations 1 and 2.

[Reaction Scheme 1]

[Reaction Scheme 2]

According to equations (1) and ( ₂ ), hydrogen peroxide (H ₂ O ₂ ) is produced when glucose contained in blood reacts with glucose oxidase, and ABTS changes from colorless to green while oxidizing by reaction with hydrogen peroxide, The higher the degree of oxidation, the darker the green.

The absorbent member 170 may further include a blood glucose absorbing region at a portion of the substrate 110 connected to the reaction region 114 in a direction away from the blood glucose inflow region 112, . Part of the plasma is absorbed by the absorptive member 170 through the reaction region 114 when the inflow amount of the plasma containing the blood sugar is large, so that the plasma can be prevented from leaking out of the biochip 100 for blood glucose measurement.

The biochip 100 for measuring blood sugar according to the present embodiment may further include a first adhesive member 160 and a second adhesive member 180 interposed between the substrate 110 and the porous separator 130 .

The first adhesive member 160 may be interposed between the substrate 110 and the cap member 120 and the second adhesive member 180 may be interposed between the cap member 120 and the porous separator 130 Can be intervened.

The first adhesive member 160 is formed with a through hole 162 corresponding to the blood glucose inflow region 112, the reaction region 114 and the blood glucose absorption region 118, A blood glucose permeation port 182 may be formed to allow blood glucose to pass therethrough.

The first and second adhesive members 160 and 180 may be formed using an adhesive film or an adhesive.

Referring to FIG. 1, the blood glucose meter 1 according to the present embodiment includes a biochip 100 for measuring blood sugar and a CMOS image sensor 200.

Here, since the biochip 100 for measuring blood glucose is the same as the above-described contents, a detailed description thereof will be referred to.

The CMOS image sensor 200 (CIS, CMOS Image Sensor) is a sensor that detects an optical signal and converts it into a digital electric signal. The sensor can be implemented in a simple scanning method and a signal processing circuit can be implemented on a single chip It is possible to miniaturize the product. In addition, CMOS process technology can be used interchangeably, which can lower manufacturing cost and power consumption is very low, so it is easy to apply to products with limited battery capacity.

The principle of the CMOS image sensor 200 is as follows. There is a single photodiode in the sensor where the light is absorbed and converted to another signal. This process is based on the photoelectric effect principle. When a photon is accumulated in the form of a charge and transformed from an electron, the amount is proportional to the number of photons detected by touching the CMOS image sensor. The accumulated charges are amplified in analog voltage form and can be converted to digital numbers. The numbers displayed on the digital output are proportional to the number of photons detected by the image sensor. If there are other materials on the surface of the image sensor that interfere with the passing of the photons, the digital output number may decrease.

According to this embodiment, visible light may be used as the light source, but is not limited thereto.

The above CMOS image sensor can be mounted on a smart phone (not shown) or a camera (not shown). Particularly, when blood glucose is measured using the CMOS image sensor 200 of a smartphone, blood glucose can be measured very quickly and effectively.

Specifically, an application capable of quantitatively displaying the photon number on a smart phone equipped with a CMOS image sensor can be installed.

In addition, it is possible to obtain a sample having various blood glucose concentrations in advance to secure a relationship database between blood glucose concentration and the number of measured photons, and then apply the database to an application. When such an application is operated, when a blood sample is injected into a biochip for blood glucose measurement and reacted, the photon count detected by the CMOS image sensor can be immediately displayed as the blood glucose concentration.

It goes without saying that such an application can be embedded in a camera equipped with a CMOS image sensor.

Example

Manufacturing example  1: Preparation of reaction reagent solution

5 mg of glucose oxidase (100,000 units / g) was added to 2.5 ml of purified water to prepare a glucose oxidase solution. 5 mg of peroxidase (300 units / g) was added to 2.5 ml of purified water to prepare a peroxidase solution. ABTS 20 mg was added to 1 ml of purified water to prepare a 35 mM ABTS solution.

All solutions prepared were stored in the refrigerator until use. A glucose oxidase solution, a peroxidase solution and an ABTS solution were mixed at a volume ratio of 2: 2: 1 to prepare a reaction reagent solution.

Manufacturing example  2: Preparation of amine functionalized silica nanoparticles

6.16 ml of ammonia are mixed with 0.5 ml of aminopropyltriethoxysilane, and then 6.95 ml of ethanol and 4.48 ml of purified water are added and stirred for 1 hour. This was prepared by adding 2.5 ml of tetraethoxysilane.

Manufacturing example  3: Preparation of a mixture of amine functionalized silica nanoparticles and reaction reagent

1 mg of the amine-functionalized silica nanoparticles of Preparation Example 2 was added to 0.1 ml of the reaction reagent solution of Preparation Example 1 and mixed to prepare a mixture of the amine-functionalized silica nanoparticles immobilized with the reaction reagent and the reaction reagent.

Amine-functionalized silica nanoparticles play an important role in the biochip for blood glucose measurement. Usually silica nanoparticles have chemical stability, light transparency, low temperature encapsulation, thermal stability, and biocompatibility. In particular, glucose oxidase and chromogens can be immobilized on an amine functionalized silica substrate with an electrostatic attraction.

Manufacturing example  4: Manufacture of biochip for blood glucose measurement

A method of manufacturing a biochip for blood glucose measurement according to the present invention will be described with reference to FIGS.

First, a hydrophilic PET (polyethylene terephthalate) film, a sample pad made of a glass fiber membrane, a plasma separation membrane, a double-sided adhesive tape, and an absorbent pad were prepared.

A circular area of a detection area A having a diameter of 4 mm and a depth of 0.2 mm was formed on the reaction area 114 using a press at the center of a substrate 110 made of a hydrophilic PET film (width x length x thickness = 24 x 8 x 0.1 mm) Thereby forming a concave portion 116 of the light emitting diode.

Next, 15 μl of a mixture (140) of the amine-functionalized silica nanoparticles fixed with the reagent according to Production Example 3 and the analysis reagent was supplied to the reaction region 114 on the substrate 160, Coated in an oval shape, and stored in a refrigerator until use.

The four corners of the double-sided tape 1 (width x length x thickness = 24 x 8 x 0.1 mm) 160 having the through-hole 162 at the center were arranged on the substrate so as to coincide with the four corners of the substrate 110.

Thereafter, a circular convex portion 122 having a diameter of 3 mm and a height of 0.2 mm was formed at the center of a top layer (width x width x thickness = 10 x 8 x 0.1 mm) 120 made of a hydrophilic PET film using a press , A space is formed by disposing a top layer 120 on a substrate 110 in such a direction that a concave portion of the substrate and a convex portion of the top layer are separated from each other, So that the substrate 110 and the top layer 120 are joined. Where the space forms the detection area A of the photon.

(Width x length x thickness x 8 x 8 x 0.1 mm) 170 made of cellulose fiber having a high absorption capacity is connected to the reaction region 114 in a direction away from the blood glucose inflow region 112 of the substrate 110 The blood glucose absorbing region 118 was adhered to the end on the substrate on which the blood glucose absorbing region 118 was formed to absorb excess plasma.

Next, one end of the plasma separation membrane (width x length x thickness = 6 x 8 x 0.1 mm) 130 is overlapped with one end of the top layer 120, and a plasma separation membrane is formed between the top layer and the double- Sided adhesive tape 2 (width x length x thickness = 6 x 8 x 0.1 mm). Where the edges of the plasma separation membrane and the edges of the substrate coincide.

The double-sided pressure-sensitive adhesive tape 2 was formed with a blood-glucose penetration hole having a diameter of 3 mm at its center. At this time, the sample pad 150 and the absorption pad 170 are adhered to each other so as to coincide with the corners of the substrate 110 to prevent leakage of the blood sample and serve as a packing. The double-sided pressure-sensitive adhesive tapes 1 and 2 were each formed with a through-hole at the center so as not to interfere with blood glucose permeation and light transmission.

Test Example : Blood glucose concentration With photon number  Relationship analysis

4 is a photograph comparing the biochip for pre-reaction blood glucose measurement and the post-reaction biochip for glucose measurement in blood glucose measurement using four blood samples (sample 2, sample 7, sample 14 and sample 24 from left) having different blood glucose concentrations .

Referring to FIG. 4, it can be seen that as the blood glucose concentration increases, the green becomes thicker.

Blood containing various concentrations of blood glucose was collected at different time intervals after feeding ipGTT from mice and high fat diet mice (HFD). Samples were collected from mice after 5 hours and 20 hours of fasting, and were also collected at 0, 15, 30, 60, 120, and 140 minutes after feeding. The samples were also collected from high fat diet mice at 0, 30, 90, 120 and 160 minutes to prepare 28 blood samples.

The blood glucose concentration of the prepared samples was measured by a GM 9 blood glucose analyzer and an Accu-check point-of-care blood glucose sensor. The 28 samples of blood were injected into the biochip for blood glucose measurement of the present invention, The number of photons passing through the biochip for blood glucose measurement was measured by a CMOS image sensor. For photon analysis, NOON0PC30L, 110000 active single chip CMOS image sensor was used.

The concentrations of blood glucose and the results of the photon number analysis are shown in the graphs of Table 1 and FIG. 5 below.

Sample Blood glucose concentration (占 퐂 / ml) Number of photons by CMOS sensor
(AU) GM9 Accu-Check One 110 102 161 2 131 126 159 3 163 153 146 4 178 149 143 5 183 165 143 6 193 178 139 7 246 216 121 8 249 183 119 9 262 258 115 10 269 283 115 11 291 301 106 12 308 261 100 13 316 321 94 14 321 299 87 15 330 317 85 16 366 369 73 17 378 367 70 18 380 351 67 19 388 385 58 20 416 450 50 21 437 421 45 22 443 427 44 23 448 469 45 24 452 415 45 25 472 467 37 26 491 515 36 27 494 484 36 28 586 548 31

 Referring to Table 1 and FIG. 5, as the blood glucose concentration increases, the measured photon number decreases. This is because as the concentration of blood glucose increases, the color of the photodetection area of the biochip for blood glucose measurement becomes thicker, thereby lowering the light transmittance.

In FIG. 6, the expected value of the abscissa indicates the blood glucose concentration of GM9, and the ordinate indicates the blood glucose concentration of the Accu-Check device. In Fig. 7, the blood glucose concentration on the abscissa is a blood glucose concentration measurement value measured by GM9.

6 and 7, the graph linearity of the blood glucose concentration between GM9 and Accu-Check is 0.9712, and the graph linearity with respect to the photon number is 0.9663. When the experimental results are database, By measuring the number of photons by dropping blood on a biochip for blood glucose measurement, the blood glucose of the blood can be measured in real time. Using this in a smartphone application, blood glucose measurement can be very simple and accurate.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, many modifications and changes may be made by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims. The present invention can be variously modified and changed by those skilled in the art, and it is also within the scope of the present invention.

1: blood glucose meter 100: biochip for blood glucose measurement
110: substrate 120: cap member
130: porous separator 140: silica nanoparticle and reaction reagent
150: pad 160: first adhesive layer
170: absorbing member 180: second adhesive layer
200: CMOS image sensor

Claims (24)

A substrate on which a blood glucose inflow region into which blood glucose flows into the first surface and a reaction region connected with the blood glucose inflow region are formed, and a recess is formed in the reaction region;
A porous separator disposed on the blood-glucose inflow area opposite to the one surface of the substrate; And
A silica nanoparticle and a reaction reagent distributed on the reaction region of the substrate;
A biochip for blood sugar measurement. The method according to claim 1,
Wherein the biochip for blood glucose measurement further comprises a cap member on the reaction region. 3. The method of claim 2,
Wherein the cap member includes a convex portion, and the convex portion and the concave portion are coupled with each other in a direction in which the convex portion of the convex portion and the concave portion of the concave portion are apart from each other to form a space. Biochip. The method according to claim 1,
Wherein the concave portion accommodates blood sugar and forms a photon detection region. The method of claim 3,
Wherein the space accommodates blood glucose and forms a photon detection region. The method according to claim 1,
Wherein the porous separator is selected from the group consisting of nitrocellulose, polysulfone, polyester, polyamide, polyimide, polyamideimide, poly (meta-phenylene isophthalamide), polyetherketone, polyetherimide, For example, made of polypropylene, polypropylene, polyvinylidene fluoride, perfluoropolymers, polyvinyl chloride, polyvinylidene chloride, polyethylene glycol derivatives, polyoxides, polyvinylacetates, polystyrenes, polyacrylonitriles, polymethylmethacrylates and nonwoven fibers Wherein the biochip comprises at least one selected from the group consisting of: The method according to claim 1,
Wherein the silica nanoparticles are hydrophilic functional group functionalized with a hydrophilic functional group. 8. The method of claim 7,
Wherein the hydrophilic functional group is at least one selected from the group consisting of an amine group, a carboxyl group, a hydroxyl group, and a thiol group. The method according to claim 1,
Wherein the silica nanoparticles have a diameter of 10 to 300 nm. The method according to claim 1,
Wherein the reaction reagent comprises glucose oxidase and chromogen. 2. The biochip of claim 1, wherein the reaction reagent comprises glucose oxidase and chromogen. 11. The method of claim 10,
Wherein the glucose oxidase comprises glucose oxidase immobilized on the silica nanoparticles. 2. The biochip as claimed in claim 1, wherein the glucose oxidase is glucose oxidase. 11. The method of claim 10,
The above chromogens are preferably selected from the group consisting of 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid), O- O-Dianisidine, O-Toluidine-Blue, Potassium Iodide, Tetra methyl benzidine, 8-anilino-1-naphthalenesulfonic acid ammonium binding- [3-methyl-2-benzothiazolinone hydrazone] N-sulfonylbenzenesulfonate monosodium combined with 8-anilino-1-naphthalene sulfonic acid (Meta [3-methyl-2-benzothiazolinonehydrazone] N-sulfonyl benzenesulfonate monosodium ammonium, MBTHSB-ANS), 3,3 ', 5,5'-tetramethylbenzidine and syringaldazine, 3,3', 5,5'-tetramethylbenzidine and syringaldazine, Primaquine diphosphate, Wherein the biochip is one selected from the group consisting of thiazole yellow G and Auramine O-anhydrous. 11. The method of claim 10,
Wherein the reaction reagent further comprises peroxidase (Peroxidase). The method according to claim 1,
The substrate further forms a blood glucose absorbing region connected to the reaction region in a direction away from the blood glucose inflow region,
Wherein the biochip for blood glucose measurement further comprises an absorbing member on the blood glucose absorbing region. The method according to claim 1,
Wherein the substrate and the cap member are light transmissive. The method according to claim 1,
Wherein at least one of the substrate and the cap member is formed of a material selected from the group consisting of polyester, polyacrylate, polyethylene (PE), polypropylene (PP), polysulfone (PS), polyethersulfone (PES), polycarbonate , Polyarylate (PAR), polyimide (PI), polyamide, and glass. 15. The method of claim 14,
The biochip for blood glucose measurement comprises:
At least one member selected from the group consisting of a first adhesive member between the substrate and the cap member, and a second adhesive member between the cap member and the porous separator,
Wherein the first adhesive member has through-holes corresponding to the blood-glucose inflow region, the reaction region, and the blood-glucose absorption region,
Wherein the second adhesive member is formed with a blood-sac-through opening corresponding to the blood-glucose inflow area so that blood glucose passes therethrough. 18. The method of claim 17,
Wherein one end of the cap member does not overlap with all or a part of the blood glucose perforation hole of the second adhesive member. The method according to claim 1,
The biochip for blood glucose measurement further comprises a pad member on the porous separation membrane. A substrate on which a blood glucose inflow region into which blood glucose flows into the first surface and a reaction region connected with the blood glucose inflow region are formed, and a recess is formed in the reaction region; A porous separator disposed on the blood-glucose inflow area opposite to the one side of the substrate; And a reaction reagent for reacting with the silica nanoparticles distributed on the reaction region of the substrate. And
A CMOS image sensor for measuring the number of photons transmitted through the biochip for blood glucose measurement;
Includes a blood glucose meter. 21. The method of claim 20,
Wherein the transmitted photon count is a photon number transmitted through the concave portion as the photon detection region of the biochip for measuring blood glucose. 21. The method of claim 20,
Wherein the CMOS image sensor is mounted on a smartphone. 23. The method of claim 22,
Wherein the smartphone is provided with an application for quantitatively displaying the photon number measured by the CMOS image sensor. 24. The method of claim 23,
Wherein the application quantitatively displays the concentration of blood glucose based on a database in which the relationship between the concentration of blood glucose and the photon number transmitted through the biochip for measuring blood sugar is measured.

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