A dry chemical reagent tablet for quantitatively detecting glucose and a preparation method thereof

Abstract

The present invention provides a dry chemical reagent sheet for quantitatively detecting glucose and a preparation method thereof, and relates to the field of dry chemical reagent sheets. The dry chemical reagent sheet includes a light-transmitting layer, a reagent layer and a diffusion layer, wherein the reagent layer is arranged above the light-transmitting layer, the diffusion layer is arranged above the reagent layer, the diffusion layer includes a diffusion layer reagent or is formed by a diffusion layer reagent, and the diffusion layer reagent includes a hydrophilic polymer, a surfactant, and a particulate matter, wherein the particulate matter includes a first particulate matter and a second particulate matter; the number ratio of the first particulate matter to the second particulate matter is 1:(0.25-4), and the difference in particle size between the first particulate matter and the second particulate matter is 1-4 μm. The dry chemical reagent sheet provided by the present invention can quickly and accurately quantitatively detect the content of glucose in a liquid sample, has good repeatability, and the preparation process of the dry chemical reagent sheet is simple, the sample volume is small, and it is easy to store and transport.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] The present invention claims the priority of Chinese patent application CN 202411436952.8 filed on October 15, 2024, entitled “A dry chemical reagent sheet for quantitative detection of glucose and its preparation method”, and the entire contents of the above application are incorporated herein by reference. Technical Field

[0003] The invention relates to the field of dry chemical reagent tablets, and in particular to a dry chemical reagent tablet for quantitatively detecting glucose and a preparation method thereof. Background Art

[0004] Glucose is a vital energy source for the human body and is the energy source for human cells. However, the wet method commonly used in clinical practice usually requires a large number of manual steps such as sample preparation, reagent addition, and reaction monitoring. The operation is complicated and has a high risk of error. At the same time, the detection process usually takes a long time to complete and is relatively costly, so its application has always been relatively limited.

[0005] Quantitative dry film testing belongs to dry chemical analysis, which is a technical method widely used in clinical testing and biochemical research. Its core feature is to pre-dry a variety of reaction reagents and fix them on paper or other carriers. Then, during the actual test, the liquid in the sample (such as whole blood, serum, plasma, urine, etc.) is used as the reaction medium to make the components to be tested in the sample react directly with the dry reagent. This reaction usually produces a color change, and qualitative or quantitative analysis is performed through naked eye observation or instrument detection. Compared with traditional wet chemistry, quantitative dry film testing has many advantages such as simple and convenient operation, fast results, low cost, small dosage, easy storage, easy to combine with automated equipment, strong adaptability, and environmental friendliness.

[0006] To this end, we hope to develop a glucose quantitative detection dry sheet that can not only effectively simplify the glucose detection process and reduce the cost of use, but also optimize the application environment and achieve convenient and fast POCT (point-of-care testing). Summary of the invention

[0007] In view of the above analysis, the present invention aims to provide a dry chemical reagent sheet for quantitative detection of glucose and a preparation method thereof, so as to solve the problems in the prior art such as large blood drawing volume, long analysis cycle, complicated process, and inability to measure in real time.

[0008] The purpose of the present invention is mainly achieved through the following technical solutions:

[0009] In the first aspect, the present invention provides a dry chemical reagent sheet for quantitative detection of glucose, comprising a light-transmitting layer, a reagent layer and a diffusion layer, wherein the reagent layer is arranged above the light-transmitting layer, the diffusion layer is arranged above the reagent layer, the diffusion layer comprises a diffusion layer reagent or is formed by a diffusion layer reagent, the diffusion layer reagent comprises a hydrophilic polymer, a surfactant, and particulate matter, wherein the particulate matter comprises a first particulate matter and a second particulate matter; the number ratio of the first particulate matter to the second particulate matter is 1:(0.25-4), preferably 1:(0.5-1.5), and the difference in particle size between the first particulate matter and the second particulate matter is 1-4 μm, preferably 1.5-2.5 μm.

[0010] Preferably, the particle size of the first particles is 4 to 8 μm, preferably 4.5 to 5.5 μm.

[0011] Preferably, the particle size of the second particles is 1 to 4 μm, preferably 2.5 to 3.5 μm.

[0012] Preferably, the amount of hydrophilic polymer in the diffusion layer reagent is 10 to 25 g / m 2 , preferably 15 to 20 g / m 2 ; The dosage of surfactant is 5-10g / m 2 , preferably 7 to 9 g / m 2 ; The amount of particles is 50~500g / m 2 , preferably 100 to 200 g / m 2 .

[0013] Preferably, in the diffusion layer reagent, the hydrophilic high molecular polymer includes at least one of cellulose acetate and cellulose nitrate; preferably, the viscosity of the hydrophilic high molecular polymer is 30 to 45 poise, preferably 35 to 40 poise.

[0014] Preferably, in the diffusion layer reagent, the surfactant includes at least one of Tween, Triton and surfynol.

[0015] Preferably, in the diffusion layer reagent, the first particulate matter and the second particulate matter are the same or different, and each independently comprises at least one of titanium dioxide, barium sulfate, diatomaceous earth, kaolin, barium titanate, and polystyrene microspheres.

[0016] Preferably, the reagent layer comprises a reagent layer reagent or is formed by a reagent layer reagent, and the reagent layer reagent comprises a buffer system, peroxidase, glucose oxidase, a color developer, an adhesive, a surfactant, and a colloid.

[0017] Preferably, in the reagent layer reagent, the buffer system includes at least one of an acetate buffer system, a phosphate buffer system, a Tris buffer system, and a boric acid buffer system, the pH of the buffer system is 3 to 7, preferably 4 to 6, and the amount of the buffer system is 50 to 100 g / m 2 , preferably 70 to 80 g / m 2 .

[0018] Preferably, the amount of peroxidase in the reagent layer is 100,000 to 500,000 U / m 2 , preferably 300000~400000U / m 2 The dosage of glucose oxidase is 30000~90000U / m 2 , preferably 70000~90000U / m 2 .

[0019] Preferably, in the reagent layer reagent, the surfactant includes at least one of Tween, Triton, surfynol, and S24. Preferably, the amount of the surfactant is 20 to 50 g / m 2 , preferably 30 to 40 g / m 2 The adhesive includes at least one of glycerol, starch slurry, HPC, polyvinyl chloride, and polypropylamide. Preferably, the amount of the adhesive is 10 to 100 g / m 2 , preferably 40 to 50 g / m 2 The colloid includes at least one of gelatin, pullulan, polyvinyl alcohol, polyacrylamide, and a hydrophilic cellulose derivative. Preferably, the amount of the colloid is 100 to 300 g / m 2 , preferably 150 to 200 g / m 2 .

[0020] Preferably, in the reagent layer reagent, the color developer includes 4-aminoantipyrine and 1,5-dihydroxynaphthalene, wherein the amount of 4-aminoantipyrine is 0.1 to 3 g / m 2 , preferably 0.5 to 2 g / m 2 , the dosage of 1,5-dihydroxynaphthalene is 0.1~2g / m 2 , preferably 0.3 to 1 g / m 2 .

[0021] Preferably, the thickness of the reagent layer is 10-100 μm, the thickness of the diffusion layer is 200-800 μm, and the thickness of the light-transmitting layer is 30-300 μm.

[0022] In a second aspect, the present invention provides a method for preparing the dry chemical reagent sheet, characterized in that it comprises the following steps:

[0023] Step 1: Apply the reagent layer reagent on the light-transmitting layer and dry it;

[0024] Step 2: Apply the diffusion layer reagent on the reagent layer and dry.

[0025] Beneficial effects:

[0026] The dry chemical reagent tablet provided by the present invention is easy to operate and can quickly and accurately quantitatively detect the content of glucose in a liquid sample. The test result has high accuracy and good repeatability. The dry chemical reagent tablet has a simple preparation process, a small sample size, is easy to store and transport, has low cost, and has a wide range of applications. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is a schematic structural diagram of the dry chemical reagent tablet provided by the present invention.

[0028] Figure 2 This is a color comparison diagram of the dry chemical reagent sheet of Example 1 provided by the present invention.

[0029] In the figure, 1-upper outer frame, 2-sample loading hole of upper outer frame, 3-diffusion layer, 4-reagent layer, 5-light-transmitting layer, 6-light-transmitting hole of lower outer frame, 7-lower outer frame. DETAILED DESCRIPTION

[0030] Preferred embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings, wherein the accompanying drawings constitute a part of the present invention and are used to explain the principles of the present invention together with the embodiments of the present invention.

[0031] In recent years, due to the mutual penetration and combination of multidisciplinary high-tech in the field of dry chemistry, the analysis of dry chemical detection methods has made great progress, and dry chemical detection methods have been widely used in clinical biochemical tests. The dry chemical detection method was developed on the basis of the glucose oxidase method. Due to its simple and rapid operation, small sample volume, high precision and specificity, it is particularly suitable for emergency testing.

[0032] The term POCT refers to medical testing performed directly at the patient's side, so it is also called "bedside testing". Its main advantage is that the test can be performed anytime and anywhere, and treatment decisions can be made in a timely manner. This testing technology does not require special operators and can be used by non-testing personnel. At the same time, the use of this technology has expanded from regular wards to emergency rooms, operating rooms, ICUs, patient homes, ambulances, and accident sites, and is becoming more and more popular.

[0033] The present invention provides a dry chemical reagent sheet for rapidly measuring glucose content. The dry chemical reagent sheet is composed of an upper outer frame, a diffusion layer, a reagent layer, a light-transmitting layer and a lower outer frame from top to bottom. The diffusion layer is a porous cellulose membrane, which is composed of a cellulose solution and one or more of titanium dioxide, barium sulfate, barium titanate and polystyrene microspheres. The reagent layer includes a buffer system, a biological enzyme, a color developer, a surfactant, a colloid and the like. The diffusion layer, the reagent layer and the light-transmitting layer are bonded together in sequence through a coating process. A sample addition hole is provided at the center of the upper outer frame, and a light-transmitting hole is provided at the center of the lower outer frame. A liquid sample is dropped into the sample addition hole, and after being evenly dispersed through the diffusion layer, it reacts with a preset reagent in the reagent layer to produce a color-developing circular spot. The concentration of glucose is quantitatively determined according to the depth of the color of the circular spot.

[0034] The dry chemical reagent sheet of the present invention uses the liquid phase in the liquid sample as a solvent to cause the glucose to react chemically and colorimetrically with the reaction substrate on the dry chemical reagent sheet of the present invention, and the glucose content is determined by the depth of the color.

[0035] Hereinafter, the present invention will be described in detail.

[0036] In the first aspect, the present invention provides a dry chemical reagent sheet for quantitative detection of glucose, comprising a light-transmitting layer and a reagent layer, wherein the reagent layer is arranged above the light-transmitting layer, the reagent layer comprises a reagent layer reagent or is formed by a reagent layer reagent, and the reagent layer reagent comprises a buffer system, a biological enzyme, a color developer, an adhesive, a surfactant, and a colloid.

[0037] As a specific embodiment of the present invention, in the reagent layer reagent, the buffer system is one of an acetate buffer system, a phosphate buffer system, a Tris buffer system, and a boric acid buffer system, and the pH is 3 to 7, preferably 4 to 6. The amount of the buffer system is 50 to 100 g / m 2 , preferably 70 to 80 g / m 2 .

[0038] As a specific embodiment of the present invention, in the reagent layer reagent, the biological enzyme includes peroxidase and glucose oxidase, wherein:

[0039] The dosage of peroxidase is 100000~500000U / m 2 , preferably 300000~400000U / m 2 ,

[0040] The dosage of glucose oxidase is 30000~90000U / m 2 , preferably 70000~90000U / m 2 .

[0041] As a specific embodiment of the present invention, in the reagent layer reagent, the activity of peroxidase is 250-300 U / mg, preferably 270-275 U / mg, and the activity of glucose oxidase is 250-300 U / mg, preferably 278-284 U / mg.

[0042] As a specific embodiment of the present invention, in the reagent layer reagent, the color developer includes 4-aminoantipyrine and 1,5-dihydroxynaphthalene. The amount of 4-aminoantipyrine is 0.1 to 3 g / m 2 , preferably 0.5 to 2 g / m 2 ; The dosage of 1,5-dihydroxynaphthalene is 0.1~2g / m 2 , preferably 0.3 to 1 g / m 2 .

[0043] As a specific embodiment of the present invention, in the reagent layer reagent, the surfactant is at least one of Tween (model may be Teween-80), Triton (model may be TritonX-100), surfynol (model may be surfynol 485), S24 (model may be Surfactant 10G). The amount of the surfactant used is 20 to 50 g / m 2 , preferably 30 to 40 g / m 2 .

[0044] As a specific embodiment of the present invention, the adhesive is at least one of glycerol, starch slurry, HPC, polyvinyl chloride, and polypropylamide. The amount of the adhesive is 10 to 100 g / m 2 , preferably 40 to 50 g / m 2 .

[0045] As a specific embodiment of the present invention, the colloid is at least one of gelatin, pullulan, polyvinyl alcohol, polyacrylamide, and hydrophilic cellulose derivatives. The amount of the colloid is 100 to 300 g / m 2 , preferably 150 to 200 g / m 2 .

[0046] As a specific embodiment of the present invention, in the reagent layer reagent, the gelatin is selected from at least one of photographic gelatin and edible gelatin.

[0047] As a specific embodiment of the present invention, the reagent layer has a wet film thickness of 100 to 600 μm, a dry film thickness of 10 to 100 μm, and preferably a dry film thickness of 15 to 30 μm.

[0048] As a specific embodiment of the present invention, the reagent sheet further includes a diffusion layer disposed above the reagent layer, the diffusion layer includes a diffusion layer reagent or is formed by a diffusion layer reagent, and the diffusion layer reagent includes a hydrophilic polymer, a surfactant, and particulate matter.

[0049] As a specific embodiment of the present invention, in the diffusion layer reagent, the hydrophilic polymer is selected from at least one of cellulose acetate and cellulose nitrate, wherein the amount of the hydrophilic polymer is 10 to 25 g / m 2 , preferably 15 to 20 g / m 2 .

[0050] As a specific embodiment of the present invention, in the diffusion layer reagent, the viscosity of the hydrophilic high molecular polymer is 30 to 45 poise, for example, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 poise, etc., preferably 35 to 40 poise. It should be noted that the viscosity determination method of the hydrophilic high molecular polymer such as cellulose acetate is to determine the intrinsic viscosity of cellulose acetate by the viscosity method, and the intrinsic viscosity of cellulose acetate in tetrahydrofuran solvent is determined by using the viscosity method, that is, the constant in the molecular weight equation.

[0051] As a specific embodiment of the present invention, in the diffusion layer reagent, the surfactant is at least one of Tween (model may be Teween-80), Triton (model may be TritonX-100), and surfynol (model may be surfynol 485). The amount of the surfactant used is 5 to 10 g / m 2 , preferably 7 to 9 g / m 2 .

[0052] As a specific embodiment of the present invention, the particles include first particles and second particles, the first particles are the same as or different from the second particles, and are independently selected from at least one of titanium dioxide, barium sulfate, diatomaceous earth, kaolin, barium titanate, and polystyrene microspheres; the amount of the particles is 50 to 500 g / m 2 , preferably 100 to 200 g / m 2 .

[0053] As a specific embodiment of the present invention, the average particle size of the first particles is 4 to 8 μm, for example, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5 μm, etc., preferably 4.5 to 5.5 μm, and more preferably 5 μm.

[0054] As a specific embodiment of the present invention, the average particle size of the second particulate matter is 1 to 4 μm, for example, 1.5, 2.0, 2.5, 3.0, 3.5 μm, etc., preferably 2.5 to 3.5 μm, and more preferably 3 μm.

[0055] As a specific embodiment of the present invention, the number ratio of the first particles to the second particles is 1:(0.25-4), preferably 1:(0.5-1.5), and more preferably 1:1.

[0056] As a specific embodiment of the present invention, the difference in particle size between the first particle and the second particle is 1-4 μm, such as 1.5, 2.0, 2.5, 3.0, 3.5 μm, etc., preferably 1.5-2.5 μm. It should be noted that the difference in the present invention refers to the absolute value.

[0057] In the present invention, the combination of the first particulate matter and the second particulate matter will change the pore size of the porous cellulose membrane of the diffusion layer, thereby further increasing the porosity of the diffusion layer, and further increasing the diffusion speed of the droplets on the diffusion layer and the uniformity of the diffusion circular spots. It should be noted that in the present invention, unless otherwise specified, the particle size of the particulate matter is the average particle size.

[0058] As a specific embodiment of the present invention, the diffusion layer has a wet film thickness of 500 to 1500 μm, a dry film thickness of 200 to 800 μm, and preferably a dry film thickness of 400 to 600 μm.

[0059] As a specific embodiment of the present invention, the light-transmitting layer of the dry chemical reagent sheet is a transparent plastic substrate. Such as polyethylene PE, polypropylene PP, polyester methyl methacrylate PMMA, polystyrene PS, polycarbonate PC, polyethylene terephthalate PET, etc., with PET being the best. The thickness of the light-transmitting layer is 30 to 300 μm, preferably 100 to 200 μm. The light transmittance to visible light of 540 nm wavelength is more than 80%, preferably more than 95%. Both the incident light source and the reflected light can pass through the light-transmitting layer.

[0060] As a specific embodiment of the present invention, the surface of the light-transmitting layer is pre-treated by corona, plasma irradiation, ultraviolet irradiation, etc.

[0061] As a specific embodiment of the present invention, the reagent sheet further comprises an upper outer frame, which is arranged above the diffusion layer. A sample addition hole is arranged at the center of the upper outer frame, and the diameter of the hole is 5 to 10 mm.

[0062] As a specific embodiment of the present invention, the reagent sheet further comprises a lower outer frame, which is arranged below the light-transmitting layer. A light-transmitting hole is arranged at the center of the lower outer frame, and the diameter of the hole is 5 to 10 mm.

[0063] As a specific embodiment of the present invention, a glue layer is attached to the lower outer frame for bonding the upper outer frame. The upper outer frame and the lower outer frame can fix the multi-layer film structure and prevent the oxidation of the pre-set reagent in the reagent layer, thereby extending the storage time of the reagent.

[0064] The detection principle of the dry chemical reagent sheet provided by the present invention is: a drop of patient sample is dropped on the dry sheet and evenly distributed to the reagent layer below through the diffusion layer. Under the catalysis of glucose oxidase, the glucose in the sample is oxidized to produce hydrogen peroxide and gluconate. Then the peroxidase catalyzes the dye precursor hydrogen peroxide to undergo an oxidative coupling reaction to develop color.

[0065] In the present invention, compared with the wet chemical detection method, the dry chemical reagent sheet has the advantages of fast speed, high accuracy, simple and convenient operation, and reliable results, and is an ideal method for glucose detection. It can be widely used in multiple fields such as in vitro diagnosis, drug synthesis and screening, environmental monitoring and protection, health quarantine, and forensic identification.

[0066] The dry chemical reagent tablets involved in the present invention can be used to quickly determine and analyze the content of glucose in liquid samples. The operation is simple, the detection is fast, and the accuracy is high. It is suitable for in vitro diagnosis, drug synthesis and screening, environmental monitoring and protection, health quarantine, forensic identification and other fields.

[0067] In a second aspect, the present invention provides a method for preparing a dry chemical reagent sheet for quantitatively detecting glucose, comprising the following steps:

[0068] Step 1: Apply the reagent layer reagent on the light-transmitting layer and dry it;

[0069] Step 2: Apply the diffusion layer reagent on the reagent layer and dry;

[0070] Step 3: Assemble the upper outer frame and the lower outer frame.

[0071] As a specific embodiment of the present invention, the structure of the dry chemical reagent sheet provided by the present invention can be as follows: Figure 1 shown. Figure 1 In the figure, 1 is an upper outer frame, 2 is a sample loading hole of the upper outer frame, 3 is a diffusion layer, 4 is a reagent layer, 5 is a light-transmitting layer, 6 is a light-transmitting hole of the lower outer frame, and 7 is a lower outer frame.

[0072] The preferred embodiments of the present invention are described in detail below to illustrate the principles of the present invention, but are not intended to limit the scope of the present invention.

[0073] It should be noted that, in the present invention, the amount of each component in the reagent layer reagent and the diffusion layer reagent is the amount of the listed substances, and does not include the amount of the solvent for dissolving the component.

[0074] Unless otherwise specified, the reagents used in the embodiments of the present invention are all commercially available products.

[0075] Example 1

[0076] This embodiment provides a dry chemical reagent sheet for quantitative determination of glucose and a preparation method thereof. The dry chemical reagent sheet is composed of an upper outer frame 1, a diffusion layer 3, a reagent layer 4, a light-transmitting layer 5 and a lower outer frame 7 from top to bottom (eg, Figure 1 A sample adding hole 2 is provided at the center of the upper outer frame, and a light transmission hole 6 is provided at the center of the lower outer frame.

[0077] Use Figure 1 The dry chemical reagent sheet structure shown in the figure has the following specific operation steps:

[0078] The light-transmitting layer 5 is made of PET film with a thickness of 125 μm and a light transmittance of more than 95% at a wavelength of 540 nm. The formulation of the reagent layer 4 is shown in Table 1. The prepared reagent layer coating liquid is evenly coated on the light-transmitting layer PET film, with a wet film thickness of 300 μm. Place it in a blast drying oven at 37°C, and continue to coat the diffusion layer 3 after it is completely dried, with a wet film thickness of 500 μm. The diffusion layer formulation is shown in Table 2. After drying in a fume hood, cut into 1 cm 2 The reagent sheet is assembled by using a plastic sheet as the upper outer frame 1 and the lower outer frame 7 of the reagent sheet. The central opening diameter of the upper and lower outer frames is 10 mm.

[0079] Table 1 Reagent layer formula

[0080]

[0081]

[0082] Table 2 Diffusion layer formula

[0083]

[0084] Example 2

[0085] This embodiment is basically the same as embodiment 1, except that the amount of peroxidase in the reagent layer is 176500 U / m 2 .

[0086] Example 3

[0087] This embodiment is basically the same as Embodiment 1, except that the colloid in the reagent layer is edible gelatin.

[0088] Example 4

[0089] This embodiment is basically the same as embodiment 1, except that the buffer in the reagent layer is PBS buffer.

[0090] Example 5

[0091] This embodiment is substantially the same as the embodiment 1, except that the viscosity of the cellulose acetate in the diffusion layer is 30 poise.

[0092] Example 6

[0093] This embodiment is basically the same as the embodiment 1, except that the particles in the diffusion layer are barium sulfate (the ratio of the number of barium sulfate with particle sizes of 5 μm and 3 μm is 1:1).

[0094] Example 7

[0095] This embodiment is basically the same as embodiment 1, except that the surfactant in the diffusion layer is Tween-80.

[0096] Example 8

[0097] This embodiment is basically the same as embodiment 1, except that the amount of peroxidase in the reagent layer is 176500 U / m 2 , the dosage of glucose oxidase is 39700U / m 2 .

[0098] Example 9

[0099] This embodiment is substantially the same as the embodiment 1, except that the viscosity of the cellulose acetate in the diffusion layer is 45 poise.

[0100] Example 10

[0101] This embodiment is basically the same as the embodiment 1, except that titanium dioxide particles with particle sizes of 5 μm and 3 μm are added to the diffusion layer in a quantity ratio of 1:2.

[0102] Embodiment 11

[0103] This embodiment is basically the same as the embodiment 1, except that titanium dioxide particles with particle sizes of 6 μm and 2 μm are added to the diffusion layer in a quantity ratio of 1:1.

[0104] Example 12

[0105] This embodiment is basically the same as the embodiment 1, except that, during the preparation of the diffusion layer, titanium dioxide particles with particle sizes of 8 μm and 4 μm are added in a quantity ratio of 1:1.

[0106] Embodiment 13

[0107] This embodiment is basically the same as the embodiment 1, except that titanium dioxide particles with particle sizes of 5 μm and 3 μm are added to the diffusion layer in a quantity ratio of 1:5.

[0108] Comparative Example 1

[0109] This comparative example is substantially the same as Example 1, except that in the diffusion layer, the particles only include titanium dioxide with a particle size of 3 μm.

[0110] Comparative Example 2

[0111] This comparative example is substantially the same as Example 1, except that in the diffusion layer, the particles only include titanium dioxide with a particle size of 5 μm.

[0112] Experimental example

[0113] Prepare five serum samples with glucose concentrations of 35 mg / dL, 221 mg / dL, 300 mg / dL, 480 mg / dL and 600 mg / dL (among which, serum samples with concentrations of 35 mg / dL, 300 mg / dL and 600 mg / dL are commercial composite calibrators, and the other concentrations are dilution concentrations of commercial composite calibrators). 10 μL of the sample to be tested is added dropwise to the sample well 2 of the upper outer frame. The sample penetrates the reagent layer evenly and quickly through the diffusion layer, reacts at 37°C for 5 minutes, and uses an optical density meter to measure the optical density value of the colored circular spot at a wavelength of 540 nm. The test results are shown in Table 3.

[0114] Commercially available composite calibrators with concentrations of 35 mg / dL, 300 mg / dL, and 600 mg / dL were added dropwise to the Example and the Comparative Example for reaction. The coefficient of variation CV was calculated by repeating 10 times. The results are shown in Table 4.

[0115] The color comparison results of the commercially available composite calibrants with concentrations of 35 mg / dL, 300 mg / dL, and 600 mg / dL in Example 1 are as follows: Figure 2 As shown, the three round spots with glucose concentrations from low to high are uniform in size and color, and the colors change from light to dark ( Figure 2 In the figure, from top to bottom, the corresponding concentrations are 35 mg / dL, 300 mg / dL, and 600 mg / dL).

[0116] Table 3 Performance parameters of the embodiments and comparative examples

[0117]

[0118]

[0119] Table 4 Coefficient of variation CV of the embodiments and comparative examples

[0120]

[0121] It can be seen from Table 3-4 that the correlation coefficient r of Example 1 2The slope of Example 1 is 0.0026, indicating that the optical density and glucose concentration have a good linear relationship. The optical density difference between high concentration and low concentration is large, and the color difference is obvious. The coefficient of variation CV value of Example 1 is smaller than that of Comparative Examples 1-2, and the repeatability meets the requirements, and the precision is higher.

[0122] It should be noted that the embodiments described above are only used to explain the present invention and do not constitute any limitation to the present invention. The present invention has been described with reference to typical embodiments, but it should be understood that the words used therein are descriptive and explanatory words, rather than restrictive words. The present invention may be modified as specified within the scope of the claims of the present invention, and the present invention may be revised without departing from the scope and spirit of the present invention. Although the present invention described therein relates to specific methods, materials and embodiments, it does not mean that the present invention is limited to the specific examples disclosed therein, on the contrary, the present invention can be extended to all other methods and applications with the same functions.

Claims

1. A dry chemical reagent tablet for quantitative detection of glucose, characterized in that: The method comprises a light-transmitting layer, a reagent layer and a diffusion layer, wherein the reagent layer is arranged above the light-transmitting layer, the diffusion layer is arranged above the reagent layer, the diffusion layer comprises a diffusion layer reagent or is formed by a diffusion layer reagent, the diffusion layer reagent comprises a hydrophilic polymer, a surfactant, and particles, wherein the particles comprise a first particle and a second particle; The number ratio of the first particles to the second particles is 1:(0.25-4), preferably 1:(0.5-1.5), The difference in particle size between the first particulate matter and the second particulate matter is 1 to 4 μm, preferably 1.5 to 2.5 μm.

2. The dry chemical reagent sheet according to claim 1, characterized in that: The particle size of the first particles is 4 to 8 μm, preferably 4.5 to 5.5 μm; And / or, the particle size of the second particulate matter is 1 to 4 μm, preferably 2.5 to 3.5 μm.

3. The dry chemical reagent sheet according to any one of claims 1-2, characterized in that: In the diffusion layer reagent, The amount of hydrophilic polymer is 10-25g / m 2 , preferably 15 to 20 g / m 2 ; And / or, the amount of surfactant is 5 to 10 g / m 2 , preferably 7 to 9 g / m 2 ; And / or, the amount of particles is 50-500g / m 2 , preferably 100 to 200 g / m 2 .

4. The dry chemical reagent sheet according to any one of claims 1 to 3, characterized in that: In the diffusion layer reagent, the hydrophilic polymer includes at least one of cellulose acetate and cellulose nitrate; preferably, the viscosity of the hydrophilic polymer is 30 to 45 poise, preferably 35 to 40 poise; and / or, in the diffusion layer reagent, the surfactant comprises at least one of Tween, Triton, and surfynol; And / or, in the diffusion layer reagent, the first particulate matter and the second particulate matter are the same or different, and each independently comprises at least one of titanium dioxide, barium sulfate, diatomaceous earth, kaolin, barium titanate, and polystyrene microspheres.

5. The dry chemical reagent sheet according to any one of claims 1 to 4, characterized in that: The reagent layer includes or is formed by a reagent layer reagent, wherein the reagent layer reagent includes a buffer system, peroxidase, glucose oxidase, a color developer, an adhesive, a surfactant, a colloid, Preferably, the activity of peroxidase in the reagent layer reagent is 250 to 300 U / mg, preferably 270 to 275 U / mg. Preferably, in the reagent layer reagent, the activity of glucose oxidase is 250 to 300 U / mg, preferably 278 to 284 U / mg.

6. The dry chemical reagent sheet according to any one of claims 1 to 5, characterized in that: In the reagent layer reagent, the buffer system includes at least one of an acetate buffer system, a phosphate buffer system, a Tris buffer system, and a boric acid buffer system. Preferably, the pH of the buffer system is 3 to 7, preferably 4 to 6. And / or, in the reagent layer reagent, the surfactant includes at least one of Tween, Triton, surfynol, and S24, And / or, in the reagent layer reagent, the binder includes at least one of glycerol, starch slurry, HPC, polyvinyl chloride, and polypropylamide, And / or, in the reagent layer reagent, the colloid includes at least one of gelatin, pullulan, polyvinyl alcohol, polyacrylamide, and a hydrophilic cellulose derivative.

7. The dry chemical reagent sheet according to any one of claims 1 to 6, characterized in that: In the reagent layer reagent, The dosage of peroxidase is 100000~500000U / m 2 , preferably 300000~400000U / m 2 , And / or, the dosage of glucose oxidase is 30000~90000U / m 2 , preferably 70000~90000U / m 2 ; And / or, the amount of the buffer system is 50-100 g / m 2 , preferably 70 to 80 g / m 2 ; And / or, the amount of surfactant is 20 to 50 g / m 2 , preferably 30 to 40 g / m 2 ; And / or, the amount of adhesive is 10 to 100 g / m 2 , preferably 40 to 50 g / m 2 ; And / or, the amount of colloid is 100-300g / m 2 , preferably 150 to 200 g / m 2 .

8. The dry chemical reagent sheet according to any one of claims 1 to 7, characterized in that: In the reagent layer reagent, the color developing agent includes 4-aminoantipyrine and 1,5-dihydroxynaphthalene, wherein: The dosage of 4-aminoantipyrine is 0.1~3g / m 2 , preferably 0.5 to 2 g / m 2 , The dosage of 1,5-dihydroxynaphthalene is 0.1~2g / m 2 , preferably 0.3 to 1 g / m 2 .

9. The dry chemical reagent sheet according to any one of claims 1 to 8, characterized in that: The thickness of the reagent layer is 10 to 100 μm. and / or, the diffusion layer has a thickness of 200 to 800 μm; And / or, the thickness of the light-transmitting layer is 30 to 300 μm.

10. A method for preparing a dry chemical reagent sheet according to any one of claims 1 to 9, characterized in that: The steps include: Step 1: Apply the reagent layer reagent on the light-transmitting layer and dry it; Step 2: coating the diffusion layer reagent on the reagent layer and drying to obtain the dry chemical reagent sheet.

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