Dry chemical reagent tablet as well as preparation method and application thereof

Abstract

The invention provides a dry chemical reagent tablet as well as a preparation method and application thereof, and relates to the field of dry chemical reagent tablets. The dry chemical reagent piece sequentially comprises an upper outer frame, a diffusion layer, a reagent layer, a light-transmitting layer and a lower outer frame from top to bottom, the upper outer frame is provided with a sample adding hole, the lower outer frame is provided with a light-transmitting hole, the reagent layer comprises a reagent layer reagent or is formed by the reagent layer reagent, the diffusion layer comprises a diffusion layer reagent or is formed by the diffusion layer reagent, and the light-transmitting layer is formed by the diffusion layer reagent. The diffusion layer reagent comprises at least one of a hydrophilic high-molecular polymer and an adhesive, the diffusion layer reagent further comprises particulate matters, and the particulate matters comprise first particulate matters and second particulate matters; the number ratio of the first particulate matters to the second particulate matters is 1: (0.25-4); the ratio D2 / D1 of the particle size D2 of the second particulate matter to the particle size D1 of the first particulate matter is 0.2-0.8. The dry chemical reagent sheet provided by the invention is simple and convenient to operate, and can rapidly, accurately and quantitatively detect the contents of various substances in a liquid sample.

Description

Technical Field

[0001] The invention relates to the field of dry chemical reagent tablets, and in particular to a dry chemical reagent tablet and a preparation method and application thereof. Background Art

[0002] Usually, the fluids analyzed clinically include blood, urine, cerebrospinal fluid, etc. For example, the levels of triglycerides, glucose, creatinine, etc. in the blood can effectively reflect the health level of the human body.

[0003] 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.

[0004] 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.

[0005] To this end, we hope to develop a dry chemical reagent tablet that can effectively simplify the detection process of various substances in liquid samples such as blood and reduce the cost of use, while also optimizing the application environment and achieving convenient and fast POCT (point-of-care testing). Summary of the invention

[0006] In view of the above analysis, the present invention aims to provide a dry chemical reagent sheet and a preparation method and application thereof, so as to solve at least one of the following existing problems: large blood drawing volume, long analysis cycle, cumbersome process, inability to measure in real time, etc.

[0007] The purpose of the present invention is mainly achieved through the following technical solutions:

[0008] In the first aspect, the present invention provides a dry chemical reagent sheet, which includes, from top to bottom, an upper outer frame, a diffusion layer, a reagent layer, a light-transmitting layer and a lower outer frame, wherein the upper outer frame is provided with a sample loading hole, the lower outer frame is provided with a light-transmitting hole, the reagent layer includes a reagent layer reagent or is formed by a reagent layer reagent, the diffusion layer includes a diffusion layer reagent or is formed by a diffusion layer reagent, the diffusion layer reagent includes at least one of a hydrophilic polymer and an adhesive, and the diffusion layer reagent also includes 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), preferably 1:(0.5~1.5); the ratio of the particle size D2 of the second particulate matter to the particle size D1 of the first particulate matter is D2 / D1=0.2~0.8, preferably, D2 / D1=0.5~0.7.

[0009] Preferably, the first particulate matter and the second particulate matter 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 adhesive includes polyisobutylene; the hydrophilic polymer includes at least one of cellulose acetate and cellulose nitrate, and preferably, the viscosity of the hydrophilic polymer is 30 to 45 poise, preferably 35 to 40 poise.

[0010] Preferably, the amount of particles in the diffusion layer reagent is 10 to 5000 g / m 2 , preferably 100 to 1000 g / m 2 .

[0011] Preferably, the amount of the adhesive in the diffusion layer reagent is 12.5 to 75 g / m 2 , preferably 37.5 to 62.5 g / m 2 .

[0012] Preferably, the amount of the hydrophilic polymer in the diffusion layer reagent is 1 to 200 g / m 2 , preferably 10 to 100 g / m 2 .

[0013] Preferably, in the diffusion layer reagent, the particle size of the first particles is 4 to 8 μm, preferably 4.5 to 5.5 μm; the particle size of the second particles is 1 to 4 μm, preferably 2.5 to 3.5 μm.

[0014] Preferably, in the diffusion layer reagent, the particle size of the first particles is 40 to 80 μm, preferably 45 to 55 μm; the particle size of the second particles is 10 to 40 μm, preferably 25 to 35 μm.

[0015] Preferably, the diffusion layer reagent further comprises a surfactant.

[0016] Preferably, the surfactant includes at least one of Tween, Triton, surfynol, S24 and fluorosurfactant.

[0017] Preferably, the amount of the surfactant is 0.5 to 60 g / m 2 , preferably 5 to 50 g / m 2 .

[0018] Preferably, the reagent layer reagents include at least one of a buffer system, a biological enzyme, a color developer, an adhesive, a surfactant, a colloid, and an alkaline agent.

[0019] Preferably, the wet film thickness of the reagent layer is 100-600 μm, and the dry film thickness is 10-100 μm; the wet film thickness of the diffusion layer is 300-1600 μm, and the dry film thickness is 200-800 μm; and the thickness of the light-transmitting layer is 20-400 μm.

[0020] In a second aspect, the present invention provides a method for preparing the dry chemical reagent sheet, comprising the following steps:

[0021] Step 1: Apply the reagent layer reagent on the light-transmitting layer and dry it;

[0022] Step 2: Apply the diffusion layer reagent on the reagent layer and dry;

[0023] Step 3: Assemble the upper outer frame and the lower outer frame.

[0024] In a third aspect, the present invention provides a use of the dry chemical reagent sheet in quantitatively detecting a substance to be tested in a liquid sample.

[0025] Preferably, the liquid sample includes at least one of a whole blood sample, a serum sample, a urine sample and a cerebrospinal fluid sample.

[0026] Preferably, the substance to be detected in the liquid sample includes at least one of triglycerides, albumin, total protein, calcium, creatinine, creatine kinase, uric acid, glucose and cholesterol.

[0027] Beneficial effects:

[0028] The dry chemical reagent tablet provided by the present invention is easy to operate, can quickly and accurately quantitatively detect the content of various substances in liquid samples, and the detection results are highly accurate and repeatable. The dry chemical reagent tablet has a simple preparation process, a small sample size, is easy to store and transport, has a low cost, and has a wide range of applications. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is a schematic structural diagram of the dry chemical reagent tablet provided by the present invention.

[0030] 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

[0031] 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.

[0032] The present invention provides a dry chemical reagent sheet. The dry chemical reagent sheet includes 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, the reagent layer and the light-transmitting layer are sequentially bonded together through a coating process. A sample addition hole is opened at the center of the upper outer frame, and a light-transmitting hole is opened at the center of the lower outer frame. A liquid sample is dropped into the sample addition hole, and after being evenly dispersed in the diffusion layer, it reacts with a preset reagent in the reagent layer to produce a color-developing circular spot. The concentration of a substance to be tested in the liquid sample is quantitatively determined according to the depth of the color of the circular spot.

[0033] Hereinafter, the present invention will be described in detail.

[0034] First, refer to Figure 1 The present invention provides a dry chemical reagent sheet, which includes, from top to bottom, an upper outer frame 1, a diffusion layer 3, a reagent layer 4, a light-transmitting layer 5 and a lower outer frame 7, wherein the upper outer frame 1 is provided with a sample addition hole 2, and the lower outer frame 7 is provided with a light-transmitting hole 6, wherein the diffusion layer 3 is an isotropic porous diffusion membrane, the diffusion layer 3 includes a diffusion layer reagent or is formed by a diffusion layer reagent, the diffusion layer reagent includes at least one of a hydrophilic high molecular polymer and an adhesive, and the diffusion layer reagent also includes particulate matter.

[0035] As a specific embodiment of the present invention, the particulate matter includes 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 more preferably 1:1.

[0036] As a specific embodiment of the present invention, the ratio of the particle size D2 of the second particulate matter to the particle size D1 of the first particulate matter is D2 / D1=0.2-0.8, preferably, D2 / D1=0.5-0.7.

[0037] As a specific embodiment of the present invention, the first particulate matter and the second particulate matter 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 particulate matter is 10 to 5000 g / m 2 , preferably 100 to 1000 g / m 2 .

[0038] As a specific embodiment of the present invention, in the diffusion layer reagent, the adhesive is selected from polyisobutylene. Preferably, the amount of the adhesive is 12.5 to 75 g / m 2 , preferably 37.5 to 62.5 g / m 2 In the present invention, the adhesive can fix the particles so that the diffusion layer can evenly diffuse the sample.

[0039] As a specific embodiment of the present invention, the hydrophilic polymer includes at least one of cellulose acetate and cellulose nitrate. Preferably, the amount of the hydrophilic polymer is 1 to 200 g / m 2 , preferably 10 to 100 g / m 2 In the present invention, when the diffusion layer reagent includes cellulose acetate or cellulose nitrate, the diffusion layer forms a porous cellulose membrane, so that the diffusion layer evenly diffuses the sample.

[0040] 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.

[0041] As a specific embodiment of the present invention, in the diffusion layer reagent, 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; the average particle size of the second particles 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.

[0042] As a specific embodiment of the present invention, in the diffusion layer reagent, the average particle size of the first particles is 40 to 80 μm, for example, 45, 50, 55, 60, 65, 70, 75 μm, etc., preferably 45 to 55 μm, and more preferably 50 μm; the average particle size of the second particles is 10 to 40 μm, for example, 15, 20, 25, 30, 35 μm, etc., preferably 25 to 35 μm, and more preferably 30 μm.

[0043] In the present invention, the diffusion layer uses a combination of the first and second particles to change the pore size of the porous diffusion membrane of the diffusion layer, thereby further effectively improving the porosity of the diffusion layer, and further improving the diffusion speed of droplets on the diffusion layer and the uniformity of the diffusion circular spot.

[0044] In the present invention, the diffusion layer uses the combination of the first particle and the second particle to filter the macromolecular substances and interfering substances in the sample and protect the reagent layer. For example, when detecting albumin or total protein in a blood sample, the serum sample is dropped on the dry sheet, the total protein will be enriched on the first particle and the second particle, the liquid penetrates into the reagent layer, and the reagent in the reagent layer will penetrate into the diffusion layer and combine with the albumin or total protein in the sample, and the gap difference caused by the particle size control of the first particle and the second particle can more effectively ensure the uniformity and rate of serum diffusion.

[0045] It should be noted that, in the present invention, unless otherwise specified, the particle size of the particulate matter is the average particle size.

[0046] As a specific embodiment of the present invention, the diffusion layer reagent also includes a surfactant, and the surfactant is selected from at least one of Tween (model may be Teween-80), Triton (model may be TritonX-100), surfynol (model may be surfynol 485), S24 (SURFACTANT 10G), and fluorosurfactant (model may be FS-31, FS-30, FS-50, FS-60, FS-81), and the amount of the surfactant is 0.5 to 60 g / m 2 , preferably 5 to 50 g / m 2 .

[0047] As a specific embodiment of the present invention, the diffusion layer reagent may also include other required reagents such as biological enzymes, and the user may add them as needed.

[0048] As a specific embodiment of the present invention, the reagent layer includes a reagent layer reagent or is formed by a reagent layer reagent, and the reagent layer reagent includes at least one of a buffer system, a biological enzyme, a color developer, an adhesive, a surfactant, a colloid, and an alkaline agent.

[0049] In the present invention, the reagent layer is the area where the substance to be tested undergoes a chemical reaction, and can also adjust the pH value of the reagent sheet. The components of the reagent layer reagent can be selected by the user according to needs.

[0050] As a specific embodiment of the present invention, the wet film thickness of the reagent layer is 100-600 μm, and the dry film thickness is 10-100 μm.

[0051] As a specific embodiment of the present invention, the wet film thickness of the diffusion layer is 300-1600 μm, and the dry film thickness is 200-800 μm.

[0052] As a specific embodiment of the present invention, the thickness of the light-transmitting layer is 20 to 400 μm, preferably 100 to 200 μm.

[0053] As a specific embodiment of the present invention, the light-transmitting layer comprises a transparent plastic material, and the material is selected from at least one of polyethylene PE, polypropylene PP, polyester methyl methacrylate PMMA, polystyrene PS, polycarbonate PC, and polyethylene terephthalate PET, with PET being the best.

[0054] 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.

[0055] As a specific embodiment of the present invention, the diameter of the sample addition hole 2 is 5 to 15 mm.

[0056] As a specific embodiment of the present invention, the diameter of the light-transmitting hole 6 is 5 to 15 mm.

[0057] As a specific embodiment of the present invention, a glue layer is attached to the lower outer frame 7 to bond the upper outer frame 1. The upper outer frame 7 and the lower outer frame 1 can fix the multi-layer film structure and prevent the pre-set reagents in the reagent layer from oxidizing, thereby extending the storage time of the reagents.

[0058] In a second aspect, the present invention provides a method for preparing a dry chemical reagent tablet, comprising the following steps:

[0059] Step 1: Apply the reagent layer reagent on the light-transmitting layer and dry it;

[0060] Step 2: Apply the diffusion layer reagent on the reagent layer and dry;

[0061] Step 3: Assemble the upper outer frame and the lower outer frame.

[0062] In a third aspect, the present invention provides an application of a dry chemical reagent sheet for quantitatively detecting a substance to be tested in a blood sample.

[0063] As a specific embodiment of the present invention, the substance to be tested in the blood sample includes at least one of triglycerides, albumin, total protein, calcium, creatinine, creatine kinase, uric acid, glucose, and cholesterol.

[0064] 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.

[0065] 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.

[0066] Unless otherwise specified, the reagents used in the embodiments of the present invention are all commercially available products.

[0067] Example 1

[0068] This embodiment provides a dry chemical reagent tablet for quantitative determination of triglycerides and a preparation method thereof. Figure 1 The dry chemical reagent sheet structure shown in the figure has the following specific operation steps:

[0069] 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, and the wet film thickness is 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 1000 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 of the upper and lower outer frames has a diameter of 7 mm.

[0070] Table 1 Reagent layer formula

[0071]

[0072] Table 2 Diffusion layer formula

[0073]

[0074]

[0075] Example 2

[0076] 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.

[0077] Example 3

[0078] 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).

[0079] Example 4

[0080] This embodiment is basically the same as Embodiment 1, except that the surfactant in the diffusion layer is Tween-80.

[0081] Example 5

[0082] 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.

[0083] Example 6

[0084] 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 in a quantity ratio of 1:2 into the diffusion layer.

[0085] Example 7

[0086] 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 in a quantity ratio of 1:1 into the diffusion layer.

[0087] Example 8

[0088] This embodiment is basically the same as the embodiment 1, except that titanium dioxide particles with particle sizes of 8 μm and 4 μm are added in a quantity ratio of 1:1 into the diffusion layer.

[0089] Example 9

[0090] 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 in a quantity ratio of 1:5 in the diffusion layer.

[0091] Comparative Example 1

[0092] 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.

[0093] Comparative Example 2

[0094] 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.

[0095] Example 10

[0096] This embodiment provides a dry chemical reagent sheet for quantitative determination of albumin and a preparation method thereof. Figure 1 The dry chemical reagent sheet structure shown in the figure has the following specific operation steps:

[0097] 1. Preparation and coating of reagent layer

[0098] Sodium bromocresol green aqueous solution, polyvinyl alcohol (PVA) aqueous solution, polyoxyethylene lauryl ether (Brij-35) aqueous solution, fluorosurfactant FS-31 aqueous solution, tris (Tris) aqueous solution, and malic acid aqueous solution were prepared respectively, and the amount of each component was shown in Table 3. After the above solutions were mixed, the bubbles and surface foam in the mixed solvent were removed, and the coating was carried out on the support layer film of polyethylene terephthalate PET film by a coating machine, preferably the wet film coating thickness was 400 μm, and the dry film thickness of the dry film coated with the reagent layer was 250 μm.

[0099] 2. Preparation and coating of diffusion layer

[0100] Polyisobutylene (molecular weight 2400) and fluorosurfactant FS-31 were prepared into a mixed solution. Then, polystyrene microspheres with particle sizes of 50 μm and 30 μm were added in a ratio of 1:1. The amount of each component is shown in Table 4. The mixture was stirred evenly and quickly coated on the prepared dry film coated with the reagent layer. The wet film coating thickness was 500 μm and the dry film thickness was 250 μm. The dry film was dried to obtain a quantitative albumin detection dry film for in vitro diagnosis.

[0101] Table 3 Reagent layer formula

[0102]

[0103]

[0104] Table 4 Diffusion layer formula

[0105]

[0106] Embodiment 11

[0107] This embodiment is basically the same as embodiment 10, except that, during the preparation of the diffusion layer, polystyrene microspheres with particle sizes of 50 μm and 30 μm are added in a quantity ratio of 1:2.

[0108] Example 12

[0109] This embodiment is basically the same as embodiment 10, except that, during the preparation of the diffusion layer, polystyrene microspheres with particle sizes of 60 μm and 20 μm are added in a quantity ratio of 1:1.

[0110] Embodiment 13

[0111] This embodiment is basically the same as embodiment 10, except that, during the preparation of the diffusion layer, polystyrene microspheres with particle sizes of 90 μm and 50 μm are added in a quantity ratio of 1:1.

[0112] Embodiment 14

[0113] This embodiment is basically the same as embodiment 10, except that polystyrene microspheres with particle sizes of 50 μm and 30 μm are added in a quantity ratio of 1:5.

[0114] Comparative Example 3

[0115] This embodiment is basically the same as embodiment 10, except that, during the preparation of the diffusion layer, only polystyrene microspheres with a particle size of 30 μm are added as particulate matter.

[0116] Comparative Example 4

[0117] This embodiment is basically the same as embodiment 10, except that, during the preparation of the diffusion layer, only polystyrene microspheres with a particle size of 50 μm are added as particulate matter.

[0118] Embodiment 15

[0119] This embodiment provides a dry chemical reagent sheet for quantitative determination of creatine kinase and a preparation method thereof. Figure 1 The dry chemical reagent sheet structure shown in the figure has the following specific operation steps:

[0120] 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 5. The prepared reagent layer coating liquid is evenly coated on the light-transmitting layer PET film, and the wet film thickness is 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 1000 μm. The diffusion layer formulation is shown in Table 6. 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.

[0121] Table 5 Reagent layer formula

[0122]

[0123] Table 6 Diffusion layer formula

[0124]

[0125] Example 16

[0126] This embodiment provides a dry chemical reagent tablet for quantitative determination of calcium and a preparation method thereof. Figure 1 The dry chemical reagent sheet structure shown in the figure has the following specific operation steps:

[0127] 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 680 nm. The formulation of the reagent layer 4 is shown in Table 7. The prepared reagent layer coating liquid is heated and stirred at 40°C for 20 minutes, and 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 750 μm. The diffusion layer formulation is shown in Table 8. 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.

[0128] Table 7 Reagent layer reagents

[0129] Serial number Reagent layer reagent Dosage 1 Photographic Gelatin 20g / m2 2 Arsenazo III 26mmol / m2 3 SURFACTANT 10G <![CDATA[6g / m 2 ]]> 4 TritonX-100 <![CDATA[1.605g / m 2 ]]> 5 Buffer (MES buffer system) pH = 5.6, 0.012 mol / m2

[0130] Table 8 Diffusion layer reagents

[0131]

[0132] Embodiment 17

[0133] This embodiment provides a dry chemical reagent tablet for quantitative determination of creatinine and a preparation method thereof. Figure 1 The dry chemical reagent sheet structure shown in the figure has the following specific operation steps:

[0134] 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 9. The prepared reagent layer coating liquid is evenly coated on the light-transmitting layer PET film, and the wet film thickness is 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 1000 μm. The diffusion layer formulation is shown in Table 10. 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 of the upper and lower outer frames has a diameter of 7 mm.

[0135] Table 9 Reagent layer formula

[0136]

[0137] Table 10 Diffusion layer formula

[0138]

[0139] Embodiment 18

[0140] This embodiment provides a dry chemical reagent tablet for quantitative determination of uric acid and a preparation method thereof. Figure 1 The dry chemical reagent sheet structure shown in the figure has the following specific operation steps:

[0141] 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 11. The prepared reagent layer coating liquid is evenly coated on the light-transmitting layer PET film, and the wet film thickness is 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 12. 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.

[0142] Table 11 Reagent layer formula

[0143]

[0144] Table 12 Diffusion layer formula

[0145]

[0146] Embodiment 19

[0147] This embodiment provides a dry chemical reagent tablet for quantitative determination of glucose and a preparation method thereof. Figure 1 The dry chemical reagent sheet structure shown in the figure has the following specific operation steps:

[0148] 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 13. The prepared reagent layer coating liquid is evenly coated on the light-transmitting layer PET film, and the wet film thickness is 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 14. 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.

[0149] Table 13 Reagent layer formula

[0150]

[0151] Table 14 Diffusion layer formula

[0152]

[0153] Embodiment 20

[0154] This embodiment provides a dry chemical reagent tablet for quantitative determination of cholesterol and a preparation method thereof. Figure 1 The dry chemical reagent sheet structure shown in the figure has the following specific operation steps:

[0155] 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 15. The prepared reagent layer coating liquid is evenly coated on the light-transmitting layer PET film, and the wet film thickness is 200 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 16. 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 of the upper and lower outer frames has a diameter of 7 mm.

[0156] Table 15 Reagent layer formula

[0157] Serial number Reagent layer reagent Dosage 1 Photographic Gelatin <![CDATA[110g / m 2 ]]> 2 TritonX-100 <![CDATA[0.67g / m 2 ]]> 3 Buffer (Phosphate Buffer System) <![CDATA[pH=6.5,13.3g / m 2 ]]>

[0158] Table 16 Diffusion layer formula

[0159]

[0160] Test Example 1

[0161] The dry chemical reagent tablets prepared in Examples 1-9 and Comparative Examples 1-2 were tested, and five serum samples with triglyceride concentrations of 0.40mmol / L, 1.385mmol / L, 2.37mmol / L, 4.338mmol / L, and 5.65mmol / L were prepared (wherein, the serum samples with concentrations of 0.40mmol / L, 2.37mmol / L, and 5.65mmol / L were commercially available composite calibrants, and the remaining concentrations were the dilution concentrations of the commercially available composite calibrants). 5.5μL of the sample to be tested was added dropwise to the sample addition hole 2 of the upper outer frame, and the sample was uniformly and quickly infiltrated into the reagent layer through the diffusion layer, reacted at 37°C for 5min, and the optical density value of the color circle was measured at a wavelength of 540nm using an optical densitometer. The test results of the embodiments and comparative examples are shown in Table 17.

[0162] Commercially available composite calibrants with concentrations of 0.40 mmol / L, 2.37 mmol / L, and 5.65 mmol / L were added to the examples and comparative examples for reaction. The coefficient of variation CV was calculated by repeating 10 times. The results are shown in Table 18.

[0163] Table 17 Performance parameters of Examples 1-9 (S1-S9) and Comparative Examples 1-2 (D1-D2)

[0164]

[0165]

[0166] Table 18 Coefficient of variation CV of Examples 1-9 (S1-S9) and Comparative Examples 1-2 (D1-D2)

[0167]

[0168]

[0169] It can be seen from Tables 17-18 that the slope of Example 1 is 0.0846, 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 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.

[0170] Test Example 2

[0171] The dry chemical reagent sheets prepared in Examples 10-14 and Comparative Examples 3-4 were tested. Four small sheets (each sheet size 1.5 cm*1.5 cm) of the same batch of albumin quantitative detection dry sheets prepared by the present invention were taken, and serum samples with albumin standard concentrations of 13 g / L, 20 g / L, 35 g / L, 46 g / L and 62 g / L were added dropwise (wherein, serum samples with concentrations of 13 g / L, 35 g / L and 62 g / L were commercially available composite calibrators, and the remaining concentrations were dilution concentrations of commercially available composite calibrators). After waiting for 5 minutes, the optical density value was tested at a wavelength of 630 nm using an optical densitometer, wherein the corresponding optical density value also showed a gradient change relationship. The test results of the embodiments and comparative examples are shown in Table 19.

[0172] Commercially available composite calibrants with concentrations of 13 g / L, 35 g / L, and 62 g / L were added to the examples and comparative examples for reaction. The coefficient of variation CV was calculated by repeating 10 times. The results are shown in Table 20.

[0173] Table 19 Performance parameters of Examples 10-14 (S10-S14) and Comparative Examples 3-4 (D3-D4)

[0174]

[0175]

[0176] Table 20 Coefficient of variation CV of Examples 10-14 (S10-S14) and Comparative Examples 3-4 (D3-D4)

[0177]

[0178] As can be seen from Table 19 and Table 20, compared with Comparative Examples 3-4, the correlation coefficient r2 of Example 10 is 0.9868, indicating that the optical density and the albumin concentration present a good linear relationship, and the slope of Example 10 is 0.011, indicating that the optical density difference between high concentration and low concentration is large, and the color difference is obvious. In addition, the coefficient of variation CV value of Example 10 is lower than that of Comparative Examples 3-4, and the repeatability meets the requirements, and the precision is better.

[0179] Test Example 3:

[0180] The dry chemical reagent sheet prepared in Example 15 was tested, and 5 serum samples with creatine kinase concentrations of 0.04U / mL, 0.37U / mL, 0.70U / mL, 1.175U / mL, and 1.65U / mL were prepared (wherein, the serum samples with concentrations of 0.04U / mL, 0.70U / mL, and 1.65U / mL were commercially available composite calibrants, and the remaining concentrations were the dilution concentrations of commercially available composite calibrants). 11 μL of the sample to be tested was added dropwise to the upper outer frame sample well 2, and the sample was uniformly and quickly infiltrated into the reagent layer through the diffusion layer, reacted at 37°C for 5min, and the optical density value of the color circle was measured at a wavelength of 670nm using an optical density meter. The test results are shown in Table 21.

[0181] Commercially available composite calibrants with concentrations of 0.04 U / mL, 0.70 U / mL, and 1.65 U / mL were added to the examples and comparative examples to react. The coefficient of variation CV was calculated by repeating 10 times. The results are shown in Table 22.

[0182] Table 21 Performance parameters of Example 15 (S15)

[0183]

[0184] Table 22 Coefficient of variation CV of Example 15 (S15)

[0185]

[0186] Test Example 4

[0187] The dry chemical reagent sheet prepared in Example 16 was tested, and five serum samples with calcium ion concentrations of 1.8 mg / dL, 4.8 mg / dL, 9.3 mg / dL, 12.45 mg / dL, and 13.8 mg / dL were prepared (wherein, the serum samples with concentrations of 1.8 mg / dL, 9.3 mg / dL, and 13.8 mg / dL were commercially available composite calibrants, and the remaining concentrations were diluted concentrations based on commercially available composite calibrants). 10 μL of the sample to be tested was dripped into the upper outer frame sample well 2, and the sample was uniformly and quickly infiltrated into the reagent layer through the diffusion layer, reacted at 37° C. for 5 min, and the optical density value of the colored circular spot was measured at a wavelength of 680 nm using an optical densitometer. The test results of the embodiment and the comparative example are shown in Table 23.

[0188] Commercially available composite calibrators with concentrations of 1.8 mg / dL, 9.3 mg / dL, and 13.8 mg / dL were added to the Example and Comparative Example for reaction. The coefficient of variation CV was calculated by repeating 10 times. The results are shown in Table 24.

[0189] Table 23 Performance parameters of Example 16 (S16)

[0190]

[0191]

[0192] Table 24 Performance parameters of Example 16 (S16)

[0193]

[0194] Test Example 5

[0195] The dry chemical reagent sheet prepared in Example 17 was tested, and five serum samples with creatinine concentrations of 0.5 mg / dL, 0.8 mg / dL, 1.5 mg / dL, 8.5 mg / dL, and 13.2 mg / dL were prepared (wherein, the serum samples with concentrations of 0.5 mg / dL, 1.5 mg / dL, and 13.2 mg / dL were commercially available composite calibrators, and the remaining concentrations were diluted concentrations based on commercially available composite calibrators). 5.5 μL of the sample to be tested was dripped into the sample well 2 of the upper outer frame, and the sample was uniformly and quickly infiltrated into the reagent layer through the diffusion layer, reacted at 37° C. for 5 min, and the optical density value of the colored circular spot was measured using an optical densitometer at a wavelength of 540 nm. The test results of the embodiment and the comparative example are shown in Table 25.

[0196] Commercially available composite calibrators with concentrations of 0.5 mg / dL, 1.5 mg / dL, and 13.2 mg / dL were added to the Example and Comparative Example for reaction. The coefficient of variation CV was calculated by repeating 10 times. The results are shown in Table 26.

[0197] Table 25 Performance parameters of Example 17 (S17)

[0198]

[0199] Table 26 Performance parameters of Example 17 (S17)

[0200]

[0201]

[0202] Test Example 6

[0203] The dry chemical reagent sheet prepared in Example 18 was tested, and 5 serum samples with uric acid concentrations of 0.042mmol / L, 0.263mmol / L, 0.357mmol / L, 0.607mmol / L, and 0.981mmol / L were prepared (wherein, the serum samples with concentrations of 0.042mmol / L, 0.357mmol / L, and 0.981mmol / L were commercially available composite calibrants, and the remaining concentrations were the dilution concentrations of commercially available composite calibrants). 10 μL of the sample to be tested was added dropwise to the upper outer frame sample addition hole 2, and the sample was uniformly and quickly infiltrated into the reagent layer through the diffusion layer, reacted at 37°C for 5min, and the optical density value of the color circle was 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.

[0204] Commercially available composite calibrants with concentrations of 0.042 mmol / L, 0.357 mmol / L, and 0.981 mmol / L were added to the examples and comparative examples for reaction. The coefficient of variation CV was calculated by repeating 10 times. The results are shown in Table 28.

[0205] Table 27 Performance parameters of Example 18 (S18)

[0206]

[0207] Table 28 Coefficient of variation CV of Example 18 (S18)

[0208]

[0209] Test Example 7

[0210] The dry chemical reagent sheet prepared in Example 19 was tested, and five serum samples with glucose concentrations of 35 mg / dL, 221 mg / dL, 300 mg / dL, 480 mg / dL and 600 mg / dL were prepared (wherein, the serum samples with concentrations of 35 mg / dL, 300 mg / dL and 600 mg / dL were commercially available composite calibrants, and the other concentrations were diluted concentrations of commercially available composite calibrants). 10 μL of the sample to be tested was dripped into the sample well 2 of the upper outer frame, and the sample was uniformly and quickly infiltrated into the reagent layer through the diffusion layer, reacted at 37° C. for 5 min, and the optical density value of the colored circular spot was measured at a wavelength of 540 nm using an optical densitometer. The test results are shown in Table 29.

[0211] Commercially available composite calibrators with concentrations of 35 mg / dL, 300 mg / dL, and 600 mg / dL were added to the examples and comparative examples for reaction. The reaction was repeated 10 times and the coefficient of variation CV was calculated. The results are shown in Table 30.

[0212] Table 29 Performance parameters of Example 19 (S19)

[0213]

[0214] Table 30 Coefficient of variation CV of Example 19 (S19)

[0215]

[0216] Test Case 8

[0217] The dry chemical reagent sheet prepared in Example 20 was tested. Five serum samples with cholesterol concentrations of 1.80 g / L, 2.70 g / L, 3.15 g / L, 3.60 g / L, and 4.50 g / L were prepared (among which, the serum samples with concentrations of 1.80 g / L and 4.50 g / L were commercially available composite calibrants, and the other concentrations were diluted concentrations of commercially available composite calibrants). 5.5 μL of the sample to be tested was added dropwise to the sample loading hole 2 of the upper outer frame. The sample was uniformly and rapidly infiltrated into the reagent layer through the diffusion layer, reacted at 37°C for 5 min, and the optical density value of the colored circular spot was measured at a wavelength of 540 nm using an optical densitometer. The test results are shown in Table 31.

[0218] Commercially available composite calibrants with concentrations of 1.80 g / L and 4.50 g / L were added to the example and comparative example to react. The coefficient of variation CV was calculated by repeating 10 times. The results are shown in Table 32.

[0219] Table 31 Performance parameters of Example 20 (S20)

[0220]

[0221]

[0222] Table 32 Coefficient of variation CV of Example 20 (S20)

[0223]

[0224] 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 sheet, comprising, from top to bottom, an upper outer frame, a diffusion layer, a reagent layer, a light-transmitting layer and a lower outer frame, wherein: The upper outer frame is provided with a sample loading hole, and the lower outer frame is provided with a light transmission hole, wherein the reagent layer includes a reagent layer reagent or is formed by a reagent layer reagent, the diffusion layer includes a diffusion layer reagent or is formed by a diffusion layer reagent, the diffusion layer reagent includes at least one of a hydrophilic polymer and an adhesive, and the diffusion layer reagent also includes particles, wherein the particles include first particles and second particles; The number ratio of the first particles to the second particles is 1:(0.25-4), preferably 1:(0.5-1.5); The ratio of the particle size D2 of the second particles to the particle size D1 of the first particles is D2 / D1=0.2-0.8, preferably, D2 / D1=0.5-0.

7.

2. The dry chemical reagent sheet according to claim 1, characterized in that: The first particulate matter and the second particulate matter 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; and / or, the adhesive comprises polyisobutylene; And / or, 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.

3. The dry chemical reagent sheet according to claim 1 or 2, characterized in that: In the diffusion layer reagent, the amount of particles is 10 to 5000 g / m 2 , preferably 100 to 1000 g / m 2 ; And / or, in the diffusion layer reagent, the amount of the adhesive is 12.5 to 75 g / m 2 , preferably 37.5 to 62.5 g / m 2 ; And / or, in the diffusion layer reagent, the amount of the hydrophilic polymer is 1 to 200 g / m 2 , preferably 10 to 100 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 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 particles is 1 to 4 μm, preferably 2.5 to 3.5 μm.

5. The dry chemical reagent sheet according to any one of claims 1 to 3, characterized in that: In the diffusion layer reagent, the particle size of the first particles is 40 to 80 μm, preferably 45 to 55 μm; And / or, the particle size of the second particulate matter is 10 to 40 μm, preferably 25 to 35 μm.

6. The dry chemical reagent sheet according to any one of claims 1 to 5, characterized in that: The diffusion layer reagent also includes a surfactant, Preferably, the surfactant comprises at least one of Tween, Triton, surfynol, S24, and fluorosurfactant. Preferably, the amount of the surfactant is 0.5 to 60 g / m 2 , preferably 5 to 50 g / m 2 .

7. The dry chemical reagent sheet according to any one of claims 1 to 6, characterized in that: The reagent layer reagents include at least one of a buffer system, a biological enzyme, a color developer, an adhesive, a surfactant, a colloid, and an alkaline agent.

8. The dry chemical reagent sheet according to any one of claims 1 to 7, characterized in that: The wet film thickness of the reagent layer is 100-600 μm, and the dry film thickness is 10-100 μm; and / or, the diffusion layer has a wet film thickness of 300 to 1600 μm and a dry film thickness of 200 to 800 μm; And / or, the thickness of the light-transmitting layer is 20 to 400 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: Apply the diffusion layer reagent on the reagent layer and dry; Step 3: Assemble the upper outer frame and the lower outer frame.

10. Use of the dry chemical reagent sheet according to any one of claims 1 to 8 in quantitatively detecting a substance to be tested in a liquid sample, Preferably, the liquid sample comprises at least one of a whole blood sample, a serum sample, a urine sample and a cerebrospinal fluid sample; Preferably, the substance to be detected in the liquid sample includes at least one of triglycerides, albumin, total protein, calcium, creatinine, creatine kinase, uric acid, glucose and cholesterol.

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