A dry chemical reagent tablet for quantitative detection of uric acid and preparation method thereof

Abstract

The present invention provides a dry chemical reagent sheet for quantitatively detecting uric acid 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, the diffusion layer reagent includes a hydrophilic polymer, a surfactant and 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 is easy to operate, can quickly and accurately quantitatively detect the content of uric acid 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 202411438783.1 filed on October 15, 2024, entitled “A dry chemical reagent sheet for quantitative detection of uric acid and preparation method thereof”, and the entire contents of the above application are incorporated herein by reference. Technical Field

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

[0004] Uric acid is the metabolic end product of purine in human nucleic acid. A small part of the uric acid in the body is metabolized by the liver and excreted with bile, and most of it is excreted by the kidneys through urine. Under normal circumstances, the body's uric acid production rate is equivalent to the kidney excretion rate, and the serum uric acid level is basically constant. When the kidneys are damaged, uric acid excretion decreases, and uric acid retention in the body leads to increased serum uric acid levels. High uric acid levels are also one of the independent risk factors for kidney disease. Therefore, the detection and analysis of uric acid levels have important reference value in the clinical diagnosis and monitoring of kidney disease.

[0005] However, the wet method commonly used in clinical practice usually requires a large number of manual operation 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 the cost is relatively high, so its application has always been relatively limited.

[0006] 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.

[0007] To this end, we hope to develop a dry film for quantitative detection of uric acid, which can not only effectively simplify the detection process of uric acid and reduce the cost of use, but also optimize the application environment and realize convenient and fast POCT (point-of-care testing). Summary of the invention

[0008] In view of the above analysis, the present invention aims to provide a dry chemical reagent sheet for quantitative detection of uric acid 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.

[0009] The purpose of the present invention is mainly achieved through the following technical solutions:

[0010] In the first aspect, the present invention provides a dry chemical reagent sheet for quantitatively detecting uric acid, 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 first particulate matter and 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 1~4μm, preferably 1.5~2.5μm.

[0011] Preferably, the particle size of the first particles is 4 to 8 μm, preferably 4.5 to 5.5 μm.

[0012] Preferably, the particle size of the second particles is 1 to 4 μm, preferably 2.5 to 3.5 μm.

[0013] Preferably, the amount of hydrophilic polymer in the diffusion layer reagent is 10 to 200 g / m 2 , preferably 50 to 100 g / m 2 ; The dosage of surfactant is 5~50g / m 2 , preferably 10 to 20 g / m 2 ; The dosage of particles is 100~500g / m 2 , preferably 150 to 300 g / m 2 .

[0014] 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.

[0015] Preferably, in the diffusion layer reagent, the surfactant includes at least one of surfynol, Tween, S24 and Triton; preferably, the surfactant includes Triton, Tween and surfynol, wherein the mass ratio of Triton, Tween and surfynol is (0.2-0.3): (0.4-0.6): (0.4-0.6).

[0016] 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.

[0017] 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, uricase, a color developer, a surfactant and a colloid, wherein: the amount of the buffer system is 60 to 80 mol / m 2 ; The dosage of peroxidase is 3000~9500U / m 2 ; The dosage of uricase is 50~500U / m 2 ; The dosage of the developer is 0.03~0.75mmol / m 2 ; The dosage of surfactant is 0.7~5.2g / m 2 ; The amount of colloid used is 50~100g / m 2 .

[0018] Preferably, in the reagent layer reagent: the amount of the buffer system is 66 to 74 mol / m 2 ; The dosage of peroxidase is 5000~7000U / m 2 ; The dosage of uricase is 100~300U / m 2 ; The dosage of the developer is 0.2~0.5mmol / m 2 ; The dosage of surfactant is 0.7~2.5g / m 2 ; The amount of colloid used is 70~90g / m 2 .

[0019] Preferably, in the reagent layer reagent, the buffer system includes at least one of a phosphate buffer system, a Tris buffer system, and a boric acid buffer system. Preferably, the pH of the buffer system is 7.5 to 9.5, preferably 8.5 to 8.8.

[0020] Preferably, in the reagent layer reagent, the color developer includes 2-(3,5-dimethoxy-4-hydroxyphenyl)-4,5-p-(4-dimethylaminophenyl)imidazole.

[0021] Preferably, in the reagent layer reagent, the surfactant includes at least one of Tween, Triton, surfynol, S24 and glycerol. Preferably, the surfactant includes S24 and glycerol, wherein: the amount of S24 is 0.2-2 g / m 2 , preferably 0.2 to 1 g / m 2 The amount of glycerol used is 0.5 to 3.2 g / m 2 , preferably 0.5 to 1.5 g / m 2 .

[0022] Preferably, in the reagent layer reagent, the colloid includes at least one of gelatin, pullulan, polyvinyl alcohol, polyacrylamide, and a hydrophilic cellulose derivative.

[0023] Preferably, the activity of peroxidase in the reagent layer reagent is 100-500 U / mg.

[0024] Preferably, the activity of uricase in the reagent layer reagent is 1 to 100 U / mg.

[0025] 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.

[0026] 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:

[0027] Step 1: Apply the reagent layer reagent on the light-transmitting layer and dry it;

[0028] Step 2: Apply the diffusion layer reagent on the reagent layer and dry.

[0029] Preferably, in step 1, the drying temperature is 35 to 50°C, preferably 35 to 45°C.

[0030] Beneficial effects:

[0031] The dry chemical reagent tablet provided by the present invention is easy to operate and can quickly and accurately detect the content of uric acid 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

[0032] Figure 1 This is a schematic structural diagram of the dry chemical reagent tablet provided by the present invention.

[0033] Figure 2 This is a color comparison diagram of the dry chemical reagent sheet of Example 1 provided by the present invention.

[0034] 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

[0035] 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.

[0036] POCT (point-of-care testing) refers to medical testing performed directly at the patient's side, so it is also called "bedside testing". Its main advantage is that it can be tested 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 residences, ambulances, and accident sites, and is becoming more and more popular. Dry chemical reagent tablets are increasingly gaining attention as an effective method of POCT.

[0037] The present invention provides a dry chemical reagent sheet for rapidly measuring uric acid 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, polystyrene microspheres, etc. The reagent layer includes a buffer system, a biological enzyme, a color developer, a surfactant, a colloid, etc. 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 uric acid is quantitatively determined according to the depth of the color of the circular spot. 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 uric acid detection. It can be widely used in many fields such as in vitro diagnosis, drug synthesis and screening, environmental monitoring and protection, health quarantine, and forensic identification.

[0038] Hereinafter, the present invention will be described in detail.

[0039] In a first aspect, the present invention provides a dry chemical reagent sheet for quantitatively detecting uric acid, 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, a surfactant and a colloid.

[0040] As a specific embodiment of the present invention, in the reagent layer reagent, the buffer system is at least one of a phosphate buffer system, a Tris buffer system, and a boric acid buffer system, and the pH of the buffer system is 7.5 to 9.5, for example, 7.6, 7.8, 8, 8.2, 8.4, 8.6, 8.8, 9, 9.2, 9.4, preferably 8.5 to 8.8. The amount of the buffer system is 60 to 80 mol / m 2 , preferably 66 to 74 mol / m 2 .

[0041] As a specific embodiment of the present invention, the phosphate buffer system includes disodium hydrogen phosphate and sodium dihydrogen phosphate.

[0042] As a specific embodiment of the present invention, the boric acid buffer system includes boric acid and sodium borate.

[0043] As a specific embodiment of the present invention, the Tris buffer system includes Tris and hydrochloric acid.

[0044] As a specific embodiment of the present invention, in the reagent layer reagent, the biological enzymes include peroxidase and uricase, wherein: the amount of peroxidase is 3000-9500 U / m 2 , preferably 5000~7000U / m 2 ; The dosage of uricase is 50~500U / m 2 , preferably 100~300U / m 2 .

[0045] As a specific embodiment of the present invention, the activity of peroxidase is 100-500 U / mg, and the activity of uricase is 1-100 U / mg.

[0046] As a specific embodiment of the present invention, in the reagent layer reagent, the color developer is selected from 2-(3,5-dimethoxy-4-hydroxyphenyl)-4,5-p-(4-dimethylaminophenyl)imidazole, and the amount of the color developer is 0.03 to 0.75 mmol / m 2 , preferably 0.2 to 0.5 mmol / m 2 .

[0047] As a specific embodiment of the present invention, in the reagent layer reagent, the surfactant includes 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) and glycerol. The amount of the surfactant used is 0.7 to 5.2 g / m 2 , preferably 0.7 to 2.5 g / m 2 .

[0048] As a specific embodiment of the present invention, in the reagent layer reagent, the surfactant includes S24 and glycerol, wherein:

[0049] The dosage of S24 is 0.2~2g / m 2 , preferably 0.2 to 1 g / m 2 ,

[0050] The dosage of glycerol is 0.5~3.2g / m 2 , preferably 0.5 to 1.5 g / m 2 .

[0051] As a specific embodiment of the present invention, in the reagent layer reagent, the colloid is selected from at least one of gelatin, pullulan, polyvinyl alcohol, polyacrylamide, and hydrophilic cellulose derivatives; the amount of the colloid is 50 to 100 g / m 2 , preferably 70 to 90 g / m 2 .

[0052] 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. Preferably, the edible gelatin has a freezing strength of 120 g to 150 g.

[0053] In the present invention, the addition of the color developer to the reagent layer will reduce the pH of the reagent in the reagent layer, and the optimum pH of uricase is 8.5, so the pH of the buffer system is preferably between 8.5 and 8.8.

[0054] 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.

[0055] 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.

[0056] 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 200 g / m 2 , preferably 50 to 100 g / m 2 .

[0057] As a specific embodiment of the present invention, 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.

[0058] As a specific embodiment of the present invention, in the diffusion layer reagent, the surfactant is at least one of surfynol (model may be surfynol 485), Tween (model may be Teween-80), S24 (model may be Surfactant10G), and Triton (model may be TritonX-100). The amount of the surfactant used is 5 to 50 g / m 2 , preferably 10 to 20 g / m 2 .

[0059] As a specific embodiment of the present invention, in the diffusion layer reagent, the surfactant includes Triton, Tween and surfynol, and the mass ratio of Triton, Tween and surfynol is (0.2-0.3): (0.4-0.6): (0.4-0.6).

[0060] As a specific embodiment of the present invention, in the diffusion layer reagent, the particles include first particles and second particles, the first particles and the second particles are the same or different, 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 100 to 500 g / m 2 , preferably 150 to 300 g / m 2 .

[0061] 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.

[0062] 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.

[0063] 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.

[0064] 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.

[0065] 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 effectively improving the porosity of the diffusion layer, and further improving 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.

[0066] As a specific embodiment of the present invention, the diffusion layer has a wet film thickness of 300 to 1000 μm, a dry film thickness of 200 to 800 μm, and preferably a dry film thickness of 400 to 600 μm.

[0067] 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 670 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.

[0068] 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.

[0069] 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 8 to 15 mm.

[0070] 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 8 to 15 mm.

[0071] 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.

[0072] In the dry chemical reagent sheet provided by the present invention, a serum sample is dropped onto the diffusion layer of the reagent sheet and diffused into the reagent layer through the diffusion layer. In the reagent layer, uric acid generates allantoin and hydrogen peroxide under the action of uricase, and hydrogen peroxide reacts with a color developer to develop color under the action of peroxidase.

[0073] In a second aspect, the present invention provides a method for preparing a dry chemical reagent sheet for quantitatively detecting uric acid, comprising the following steps:

[0074] Step 1: Apply the reagent layer reagent on the light-transmitting layer and dry it.

[0075] Step 2: Apply the diffusion layer reagent on the reagent layer and dry;

[0076] Step 3: Assemble the upper outer frame and the lower outer frame.

[0077] As a specific embodiment of the present invention, in step 1, the drying temperature is 35-50°C, preferably 35-45°C.

[0078] 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.

[0079] 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.

[0080] 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.

[0081] Unless otherwise specified, the reagents used in the embodiments of the present invention are all commercially available products.

[0082] Example 1

[0083] This embodiment provides a dry chemical reagent sheet for quantitative determination of uric acid 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 (as shown in FIG. 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.

[0084] Use Figure 1 The dry chemical reagent sheet structure shown in the figure has the following specific operation steps:

[0085] 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 670 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.

[0086] Table 1 Reagent layer formula

[0087]

[0088] Table 2 Diffusion layer formula

[0089]

[0090] Example 2

[0091] This embodiment is substantially the same as embodiment 1, except that the pH of the buffer solution in the reagent layer is 9.01.

[0092] Example 3

[0093] This embodiment is basically the same as embodiment 1, except that the colloid in the reagent layer is edible gelatin (with a freezing strength of 150 g).

[0094] Example 4

[0095] This embodiment is basically the same as embodiment 1, except that the colloid in the reagent layer is edible gelatin (with a freezing strength of 120 g).

[0096] Example 5

[0097] This embodiment is basically the same as embodiment 1, except that, in the preparation process of the reagent sheet, the prepared reagent layer coating liquid is evenly coated on the light-transmitting layer PET film, with a wet film thickness of 300 μm. It is placed in a blast drying oven at 50°C, and after it is completely dried, the diffusion layer 3 is continuously coated, with a wet film thickness of 500 μm.

[0098] Example 6

[0099] 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.

[0100] Example 7

[0101] This embodiment is basically the same as the embodiment 1, except that the particles in the diffusion layer are anatase titanium dioxide (the ratio of the number of titanium dioxide with particle sizes of 5 μm and 3 μm is 1:1).

[0102] Example 8

[0103] This embodiment is basically the same as the embodiment 1, except that the surfactant in the diffusion layer is SURFACTANT10G (S24).

[0104] Example 9

[0105] This embodiment is basically the same as embodiment 1, except that the surfactant in the reagent layer is TritonX-100, and the dosage is 1.53 g / m 2 .

[0106] Example 10

[0107] 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.

[0108] Embodiment 11

[0109] This embodiment is basically the same as the embodiment 1, except that rutile titanium dioxide with particle sizes of 5 μm and 3 μm is added to the diffusion layer in a quantity ratio of 1:2.

[0110] Example 12

[0111] This embodiment is basically the same as the embodiment 1, except that rutile titanium dioxide with particle sizes of 6 μm and 2 μm is added to the diffusion layer in a quantity ratio of 1:1.

[0112] Embodiment 13

[0113] This embodiment is basically the same as the embodiment 1, except that, during the preparation of the diffusion layer, rutile titanium dioxide with particle sizes of 8 μm and 4 μm is added in a quantity ratio of 1:1.

[0114] Embodiment 14

[0115] This embodiment is basically the same as the embodiment 1, except that rutile titanium dioxide with particle sizes of 5 μm and 3 μm is added in a quantity ratio of 1:5.

[0116] Comparative Example 1

[0117] This comparative example 1 is substantially the same as the example 1, except that in the diffusion layer, the particles only include rutile titanium dioxide having a particle size of 3 μm.

[0118] Comparative Example 2

[0119] This comparative example is substantially the same as Example 1, except that in the diffusion layer, the particles only include rutile titanium dioxide with a particle size of 5 μm.

[0120] Experimental example

[0121] Prepare 5 serum samples with uric acid concentrations of 0.042mmol / L, 0.263mmol / L, 0.357mmol / L, 0.607mmol / L, and 0.981mmol / L (wherein, serum samples with concentrations of 0.042mmol / L, 0.357mmol / L, and 0.981mmol / L are commercially available composite calibrants, and the remaining concentrations are dilution concentrations of commercially available composite calibrants). 10 μL of the sample to be tested is added dropwise to the upper outer frame sample addition hole 2, and the sample is uniformly and quickly infiltrated into the reagent layer through the diffusion layer, reacted at 37° C. for 5 minutes, and the optical density value of the color circle is measured at a wavelength of 670nm using an optical densitometer. The test results of the embodiment and the comparative example are shown in Table 3.

[0122] Commercially available composite calibrants with concentrations of 0.042 mmol / L, 0.357 mmol / L, and 0.981 mmol / L 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.

[0123] The color comparison results of Example 1 when three concentrations (0.042mmol / L, 0.357mmol / L, 0.981mmol / L) of commercially available composite calibrants are added are as follows: Figure 2 As shown, the three circular spots with uric acid concentrations from low to high are uniform in size and color, and the colors range from light to dark ( Figure 2 In the middle, from shallow to deep, the corresponding concentrations are 0.042mmol / L, 0.357mmol / L, and 0.981mmol / L).

[0124] Table 3 Performance parameters of examples and comparative examples

[0125]

[0126]

[0127] It can be seen from Table 3 that the slope of Example 1 is 1.116, which is higher than that of Comparative Examples 1-2, indicating that the difference in optical density between high concentration and low concentration is large and the color difference is obvious; the correlation coefficient r of Example 1 is 2 It is 0.9992, which is higher than that of Comparative Examples 1-2, indicating that the correlation coefficient between uric acid concentration and optical density is higher.

[0128] Table 4 Coefficient of variation CV of the embodiments and comparative examples

[0129]

[0130]

[0131] It can be seen from Table 4 that the coefficient of variation CV of Example 1 is lower than that of Comparative Examples 1-2, indicating that the precision of Example 1 is better.

[0132] 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 uric acid, 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 particulate matter, wherein the particulate matter comprises a first particulate matter and a second particulate matter; 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 to 200 g / m 2 , preferably 50 to 100 g / m 2 ; And / or, the amount of surfactant is 5 to 50 g / m 2 , preferably 10 to 20 g / m 2 ; And / or, the amount of particles is 100-500 g / m 2 , preferably 150 to 300 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 includes at least one of surfynol, Tween, S24, and Triton; preferably, the surfactant includes Triton, Tween, and surfynol, wherein the mass ratio of Triton, Tween, and surfynol is (0.2-0.3): (0.4-0.6): (0.4-0.6); 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 a reagent layer reagent or is formed by a reagent layer reagent, and the reagent layer reagent includes a buffer system, peroxidase, uricase, a color developer, a surfactant and a colloid, wherein: The dosage of the buffer system is 60~80mol / m 2 ; And / or, the dosage of peroxidase is 3000~9500U / m 2 ; And / or, uricase dosage is 50-500 U / m 2 ; And / or, the amount of the developer is 0.03 to 0.75 mmol / m 2 ; And / or, the amount of surfactant is 0.7 to 5.2 g / m 2 ; And / or, the amount of colloid is 50-100g / m 2 ; Preferably, in the reagent layer reagent: The amount of buffer system is 66~74mol / m 2 ; And / or, the dosage of peroxidase is 5000~7000U / m 2 ; And / or, uricase dosage is 100-300 U / m 2 ; And / or, the amount of the developer is 0.2 to 0.5 mmol / m 2 ; And / or, the amount of surfactant is 0.7 to 2.5 g / m 2 ; And / or, the amount of colloid is 70-90g / m 2 .

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 a phosphate buffer system, a Tris buffer system, and a boric acid buffer system. Preferably, the pH of the buffer system is 7.5 to 9.5, preferably 8.5 to 8.8; And / or, in the reagent layer reagent, the color developer includes 2-(3,5-dimethoxy-4-hydroxyphenyl)-4,5-p-(4-dimethylaminophenyl)imidazole; And / or, in the reagent layer reagent, the surfactant includes at least one of Tween, Triton, surfynol, S24 and glycerol, preferably, the surfactant includes S24 and glycerol, wherein: the amount of S24 is 0.2-2 g / m 2 , preferably 0.2 to 1 g / m 2 The amount of glycerol used is 0.5 to 3.2 g / m 2 , preferably 0.5 to 1.5 g / m 2 ; 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 activity of peroxidase is 100 to 500 U / mg, And / or, in the reagent layer reagent, the activity of uricase is 1 to 100 U / mg.

8. The dry chemical reagent sheet according to any one of claims 1 to 7, 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.

9. A method for preparing a dry chemical reagent sheet according to any one of claims 1 to 8, 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.

10. The preparation method according to claim 9, characterized in that: In step 1, the drying temperature is 35 to 50°C, preferably 35 to 45°C.

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