CN119395004A

CN119395004A - A dry chemical reagent tablet and its preparation method and application

Info

Publication number: CN119395004A
Application number: CN202411437851.2A
Authority: CN
Inventors: 郭晓燕; 狄佐星; 严晓强; 靳海波; 何广湘; 杨索和
Original assignee: Beijing Institute of Petrochemical Technology
Current assignee: Beijing Institute of Petrochemical Technology
Priority date: 2024-10-15
Filing date: 2024-10-15
Publication date: 2025-02-07

Abstract

The present invention proposes a dry chemical reagent sheet and its preparation method and application, which relate to the field of dry chemical reagent sheets. The dry chemical reagent sheet comprises an upper outer frame, a diffusion layer, a reagent layer, a light-transmitting layer and a lower outer frame from top to bottom, wherein the upper outer frame is provided with a sample addition hole, the lower outer frame is provided with a light-transmitting hole, the reagent layer comprises a reagent layer reagent or is formed by a reagent layer reagent, the diffusion layer comprises a diffusion layer reagent or is formed by a diffusion layer reagent, the diffusion layer reagent comprises at least one of a hydrophilic polymer and an adhesive, and the diffusion layer reagent also comprises 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); 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. The dry chemical reagent sheet provided by the present invention is easy to operate and can quickly and accurately quantitatively detect the content of various substances in liquid samples.

Description

Dry chemical reagent tablet and preparation method and application thereof

Technical Field

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

Background

Generally, the fluid for clinical analysis involves blood, urine, cerebrospinal fluid and the like, and for example, the blood contains triglycerides, glucose, creatinine and the like in an amount effective to reflect the health level of the human body.

However, the wet detection commonly used in clinic generally requires a large number of manual operation steps such as sample preparation, reagent addition, reaction monitoring, etc., and is complex in operation and high in error risk, and meanwhile, the detection process generally requires a long time to complete, and has relatively high cost and has been relatively limited in application.

Quantitative dry plate detection belongs to a dry chemical analysis method, and is a technical method widely applied to clinical examination and biochemical research. The method is characterized in that various reaction reagents are dried in advance and fixed on paper sheets or other carriers, and then liquid in samples (such as whole blood, serum, plasma, urine and the like) is used as a reaction medium in actual detection, so that components to be detected in the samples directly react with the dry reagents. Such reactions typically produce a color change that is either qualitatively or quantitatively analyzed by visual inspection or instrumental detection. Compared with the traditional wet chemistry, the quantitative dry plate detection method has the advantages of simple and convenient operation, quick result, low cost, small dosage, easy storage, easy combination with automatic equipment, strong adaptability, environmental friendliness and the like.

Therefore, we hope to develop a dry chemical reagent tablet, which can not only effectively simplify the detection flow of various substances in liquid samples such as blood and reduce the use cost, but also optimize the application environment and realize convenient and quick POCT (point of care testing).

Disclosure of Invention

In view of the analysis, the invention aims to provide a dry chemical reagent tablet, a preparation method and application thereof, which are used for solving at least one of the problems of large blood drawing amount, long analysis period, complicated flow, incapability of real-time measurement and the like in the prior art.

The aim of the invention is mainly realized by the following technical scheme:

According to the first aspect, the invention provides a dry chemical reagent sheet, which 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, wherein 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 a reagent layer reagent, the diffusion layer comprises a diffusion layer reagent or is formed by a diffusion layer reagent, the diffusion layer reagent comprises at least one of a hydrophilic high polymer and a binder, the diffusion layer reagent further comprises particles, the particles comprise 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), and the ratio D2/D1=0.2-0.8 of the particle size D2 of the second particles to the first particles is preferably 0.5-0.7.

Preferably, the first particles and the second particles are the same or different, and are each independently selected from at least one of titanium dioxide, barium sulfate, diatomite, kaolin, barium titanate and polystyrene microsphere, the binder comprises polyisobutylene, the hydrophilic high molecular polymer comprises at least one of cellulose acetate and cellulose nitrate, and preferably, the viscosity of the hydrophilic high molecular polymer is 30-45 poise, and preferably, 35-40 poise.

Preferably, in the diffusion layer reagent, the particle amount is 10-5000 g/m ², preferably 100-1000 g/m ².

Preferably, the amount of the binder in the diffusion layer reagent is 12.5-75 g/m ², preferably 37.5-62.5 g/m ².

Preferably, the dosage of the hydrophilic high molecular polymer in the diffusion layer reagent is 1-200 g/m ², preferably 10-100 g/m ².

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, and the particle size of the second particles is 1 to 4 μm, preferably 2.5 to 3.5 μm.

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

Preferably, the diffusion layer reagent further comprises a surfactant.

Preferably, the surfactant comprises at least one of tween, triton, surfynol, S24, and a fluorosurfactant.

Preferably, the amount of the surfactant is 0.5-60 g/m ², preferably 5-50 g/m ².

Preferably, the reagent layer reagent comprises at least one of buffer system, biological enzyme, color developing agent, adhesive, surfactant, colloid and alkaline agent.

Preferably, the reagent layer has a wet film thickness of 100-600 μm, a dry film thickness of 10-100 μm, a diffusion layer has a wet film thickness of 300-1600 μm, a dry film thickness of 200-800 μm, and a light-transmitting layer has a thickness of 20-400 μm.

In a second aspect, the invention provides a method for preparing the dry chemical reagent tablet, which comprises the following steps:

step 1, coating a reagent layer reagent on a light-transmitting layer, and drying;

step 2, coating a diffusion layer reagent on the reagent layer, and drying;

And 3, assembling the upper outer frame and the lower outer frame.

In a third aspect, the invention provides an application of the dry chemical reagent sheet in quantitatively detecting substances to be detected 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 tested in the liquid sample comprises at least one of triglyceride, albumin, total protein, calcium, creatinine, creatine kinase, uric acid, glucose and cholesterol.

The beneficial effects are that:

The dry chemical reagent tablet provided by the invention is simple and convenient to operate, can rapidly and accurately quantitatively detect the content of various substances in a liquid sample, has high accuracy and good repeatability of detection results, and has the advantages of simple preparation process, small sample size, easiness in storage and transportation, low cost and wide application range.

Drawings

Fig. 1 is a schematic structural diagram of a dry chemical reagent tablet provided by the invention.

In the figure, a 1-upper outer frame, a 2-upper outer frame sample adding hole, a 3-diffusion layer, a 4-reagent layer, a 5-light-transmitting layer, a 6-lower outer frame light-transmitting hole and a 7-lower outer frame.

Detailed Description

Preferred embodiments of the present invention are described in detail below with reference to the attached drawing figures, which form a part of the present invention and are used in conjunction with the embodiments of the present invention to illustrate the principles of the present invention.

The invention provides a dry chemical reagent tablet. The dry chemical reagent sheet 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 diffusion layer, the reagent layer and the light-transmitting layer are sequentially bonded together through a coating process. The center of the upper outer frame is provided with a sample adding hole, and the center of the lower outer frame is provided with a light hole. And (3) dripping the liquid sample into the sample adding hole, uniformly dispersing the liquid sample through the diffusion layer, and reacting the liquid sample with a preset reagent in the reagent layer to generate a color-developing circular spot. And quantitatively determining the concentration of the substance to be detected in the liquid sample according to the shade of the color of the circular spot.

The present invention will be specifically described below.

In a first aspect, referring to fig. 1, the present invention provides a dry chemical reagent tablet, which sequentially includes 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, wherein the upper outer frame 1 is provided with a sample-adding hole 2, the lower outer frame 7 is provided with a light-transmitting hole 6, the diffusion layer 3 is a porous diffusion membrane with uniformity in all directions, 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 polymer and an adhesive, and the diffusion layer reagent further includes particles.

As a specific embodiment of the invention, the particles comprise first particles and second particles, and the quantity 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.

As a specific embodiment of the present invention, the ratio D2/d1=0.2 to 0.8, preferably, D2/d1=0.5 to 0.7, of the particle size D2 of the second particulate matter to the particle size D1 of the first particulate matter.

In one embodiment of the present invention, the first particulate matter and the second particulate matter are the same or different, and each is independently selected from at least one of titanium dioxide, barium sulfate, diatomaceous earth, kaolin, barium titanate, and polystyrene microsphere, and the amount of the particulate matter is 10-5000 g/m ², preferably 100-1000 g/m ².

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

As a specific embodiment of the present invention, the hydrophilic high molecular polymer includes at least one of cellulose acetate and cellulose nitrate. Preferably, the amount of the hydrophilic high molecular polymer is 1-200 g/m ², preferably 10-100 g/m ². In the present invention, when the diffusion layer reagent includes cellulose acetate or nitrocellulose, the diffusion layer forms a porous cellulose film so that the diffusion layer uniformly diffuses the sample.

In one embodiment of the present invention, the viscosity of the hydrophilic polymer in the diffusion layer reagent 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. The viscosity measurement method of a hydrophilic polymer such as cellulose acetate is a viscosity measurement method for measuring the intrinsic viscosity of cellulose acetate, and the intrinsic viscosity of cellulose acetate in a tetrahydrofuran solvent, that is, a constant in a molecular weight equation, is measured by using the viscosity method.

In one embodiment of the present invention, the average particle diameter of the first particles in the diffusion layer reagent 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, more preferably 5 μm, and the average particle diameter 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, more preferably 3 μm.

In one embodiment of the present invention, the average particle diameter of the first particles in the diffusion layer reagent is 40 to 80. Mu.m, for example, 45, 50, 55, 60, 65, 70, 75. Mu.m, etc., preferably 45 to 55. Mu.m, more preferably 50. Mu.m, and the average particle diameter of the second particles is 10 to 40. Mu.m, for example, 15, 20, 25, 30, 35. Mu.m, etc., preferably 25 to 35. Mu.m, more preferably 30. Mu.m.

According to the invention, the diffusion layer adopts the combination of the first particles and the second particles, so that the pore diameter of the porous diffusion membrane of the diffusion layer can be changed, thereby further effectively improving the porosity of the diffusion layer and further improving the diffusion speed of liquid drops on the diffusion layer and the uniformity of diffusion circular spots.

In the invention, the diffusion layer adopts the combination of the first particles and the second particles, so that macromolecular substances and interfering substances in a sample can be filtered, and the reagent layer is protected. For example, when albumin or total protein in a blood sample is detected, the serum sample is dropped on a dry plate, the total protein is enriched on the first particles and the second particles, the liquid permeates into the reagent layer, the reagent in the reagent layer permeates into the diffusion layer to combine with the albumin or total protein in the sample, and the gap difference caused by the particle size control of the first particles and the second particles can more effectively ensure the uniformity and the speed of serum diffusion.

In the present invention, the particle diameter of the particulate matter is an average particle diameter unless otherwise specified.

As a specific embodiment of the invention, the diffusion layer reagent further comprises a SURFACTANT, wherein the SURFACTANT is at least one selected from Tween (model number can be Teween-80), triton (model number can be Triton X-100), surfynol (model number can be surfynol), S24 (SURFACTANT 10G) and fluorine SURFACTANT (model number can be FS-31, FS-30, FS-50, FS-60 and FS-81), and the dosage of the SURFACTANT is 0.5-60G/m ², preferably 5-50G/m ².

As a specific embodiment of the present invention, the diffusion layer reagent may further comprise other desired reagents such as biological enzymes, and may be added by a user as desired.

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

In the invention, the reagent layer is a region where a substance to be detected chemically reacts, the pH value of the reagent sheet can be regulated, and the components of the reagent layer reagent can be selected by a user according to the needs.

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

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

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.

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

As a specific embodiment of the present invention, the surface of the light-transmitting layer is pretreated by corona, plasma irradiation, ultraviolet irradiation, or the like.

As a specific embodiment of the present invention, the diameter of the well 2 is 5 to 15mm.

In one embodiment of the present invention, the diameter of the light-transmitting hole 6 is 5 to 15mm.

As a specific embodiment of the present invention, a glue layer is attached to the lower frame 7 for adhering the upper frame 1. The upper and lower frames 7 and 1 can fix the multi-layered film structure and prevent the preset reagent in the reagent layer from being oxidized, and prolong the preservation time of the reagent.

In a second aspect, the invention provides a method for preparing a dry chemical reagent tablet, comprising the steps of:

step 2, coating a diffusion layer reagent on the reagent layer, and drying;

And 3, assembling the upper outer frame and the lower outer frame.

In a third aspect, the invention provides the use of a dry chemical reagent pad for quantitatively detecting a substance to be detected in a blood sample.

In one embodiment of the present invention, the substance to be tested in the blood sample includes at least one of triglyceride, albumin, total protein, calcium, creatinine, creatine kinase, uric acid, glucose, and cholesterol.

The following detailed description of the preferred embodiments of the invention illustrates the principles of the invention and is not intended to limit the scope of the invention.

In the present invention, the amounts of the respective components in the reagent layer reagent and the diffusion layer reagent are the amounts of the listed substances, and the amounts of the solvents for dissolving the components are not included.

Unless otherwise indicated, all reagents used in the examples of the present invention were commercially available.

Example 1

The embodiment provides a dry chemical reagent tablet for quantitatively determining triglyceride and a preparation method thereof. The dry chemical reagent sheet structure shown in fig. 1 is adopted, and the specific operation steps are as follows:

the light-transmitting layer 5 is made of PET film, the thickness is 125 μm, and the light transmittance at the wavelength of 540nm is more than 95%. The formulation of the reagent layer 4 is shown in Table 1, and the prepared reagent layer coating liquid is uniformly coated on the PET film of the light-transmitting layer, and the wet film thickness is 300 μm. The diffusion layer 3 was then applied after it had been completely dried in a forced air drying oven at 37℃and the wet film thickness was 1000. Mu.m. The diffusion layer formulation is shown in table 2. The reagent sheet was cut into 1cm ² pieces after drying in a fume hood, and the upper frame 1 and the lower frame 7 of the reagent sheet were assembled using a plastic sheet. The diameter of the central opening of the upper and lower outer frames is 7mm.

TABLE 1 reagent layer formulation

TABLE 2 diffusion layer formulation

Example 2

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

Example 3

This example is substantially the same as example 1 except that the particulate matter in the diffusion layer is barium sulfate (the ratio of the number of barium sulfate particles having a particle size of 5 μm to 3 μm is 1:1).

Example 4

This example is substantially the same as example 1 except that the surfactant in the diffusion layer is tween-80.

Example 5

This example is substantially the same as example 1, except that the cellulose acetate has a viscosity of 45 poise in the diffusion layer.

Example 6

This example is substantially the same as example 1 except that titanium dioxide having a particle size of 5 μm and 3 μm was added to the diffusion layer in a quantitative ratio of 1:2.

Example 7

This example is substantially the same as example 1 except that titanium dioxide having a particle size of 6 μm and 2 μm is added to the diffusion layer in a quantitative ratio of 1:1.

Example 8

This example is substantially the same as example 1 except that titanium dioxide having a particle size of 8 μm and 4 μm was added to the diffusion layer in a quantitative ratio of 1:1.

Example 9

This example is substantially the same as example 1 except that titanium dioxide having a particle size of 5 μm and 3 μm is added to the diffusion layer in a quantitative ratio of 1:5.

Comparative example 1

This comparative example is substantially the same as example 1 except that the particulate matter in the diffusion layer includes only titanium dioxide having a particle diameter of 3 μm.

Comparative example 2

This comparative example is substantially the same as example 1 except that the particulate matter in the diffusion layer includes only titanium dioxide having a particle diameter of 5 μm.

Example 10

The embodiment provides a dry chemical reagent tablet for quantitatively determining albumin and a preparation method thereof. The dry chemical reagent sheet structure shown in fig. 1 is adopted, and the specific operation steps are as follows:

1. Preparation and coating of reagent layers

Respectively preparing bromocresol green sodium aqueous solution, polyvinyl alcohol (PVA) aqueous solution, polyoxyethylene lauryl ether (Brij-35) aqueous solution, fluorine surfactant FS-31 aqueous solution, tris (Tris) aqueous solution and malic acid aqueous solution, wherein the dosage of each component is shown in table 3. After the above solutions are mixed, the bubbles and surface foam in the mixed solvent are removed, and the coating is carried out on a support layer film made of polyethylene terephthalate PET film by a coater, preferably the thickness of a wet film is 400 μm, and the dry sheet coated with the reagent layer is obtained by drying, and the dry film thickness is 250 μm.

2. Preparation and coating of diffusion layers

Polyisobutylene (molecular weight 2400) and the fluorine surfactant FS-31 are prepared into a mixed solution. Then, polystyrene microspheres with the particle sizes of 50 mu m and 30 mu m in the quantity ratio of 1:1 are added, the dosages of the components are shown in table 4, the mixture is uniformly and rapidly coated on a prepared dry sheet coated with a reagent layer, the thickness of a wet film coating is 500 mu m, the thickness of a dry film is 250 mu m, and the dry sheet is dried to obtain the albumin quantitative detection dry sheet which can be used for in-vitro diagnosis.

TABLE 3 reagent layer formulation

Table 4 diffusion layer formulation

Example 11

This example is substantially the same as example 10, except that polystyrene microspheres having a particle size of 50 μm and 30 μm were added in a 1:2 quantitative ratio during the preparation of the diffusion layer.

Example 12

This example is substantially the same as example 10, except that polystyrene microspheres having a particle size of 60 μm and 20 μm were added in a 1:1 quantitative ratio during the preparation of the diffusion layer.

Example 13

This example is substantially the same as example 10, except that polystyrene microspheres having a particle size of 90 μm and 50 μm were added in a 1:1 quantitative ratio during the preparation of the diffusion layer.

Example 14

This example is substantially the same as example 10 except that polystyrene microspheres having particle sizes of 50 μm and 30 μm are added in a quantitative ratio of 1:5.

Comparative example 3

This example is substantially the same as example 10, except that the particles were added only to polystyrene microspheres having a particle size of 30 μm during the preparation of the diffusion layer.

Comparative example 4

This example is substantially the same as example 10, except that the diffusion layer was prepared by adding only polystyrene microspheres having a particle size of 50 μm to the particles.

Example 15

The embodiment provides a dry chemical reagent tablet for quantitatively determining creatine kinase and a preparation method thereof. The dry chemical reagent sheet structure shown in fig. 1 is adopted, and the specific operation steps are as follows:

The light-transmitting layer 5 is made of PET film, the thickness is 125 μm, and the light transmittance at the wavelength of 670nm is more than 95%. The formulation of the reagent layer 4 is shown in Table 5, and the prepared reagent layer coating liquid was uniformly coated on the PET film as a light-transmitting layer, and the wet film thickness was 300. Mu.m. The diffusion layer 3 was then applied after it had been completely dried in a forced air drying oven at 37℃and the wet film thickness was 1000. Mu.m. The diffusion layer formulation is shown in table 6. The reagent sheet was cut into 1cm ² pieces after drying in a fume hood, and the upper frame 1 and the lower frame 7 of the reagent sheet were assembled using a plastic sheet. The diameter of the central opening of the upper and lower outer frames is 10mm.

TABLE 5 reagent layer formulation

TABLE 6 diffusion layer formulation

Example 16

The embodiment provides a dry chemical reagent tablet for quantitatively determining calcium and a preparation method thereof. The dry chemical reagent sheet structure shown in fig. 1 is adopted, and the specific operation steps are as follows:

The light-transmitting layer 5 is made of PET film, the thickness is 125 μm, and the light transmittance at 680nm is more than 95%. The formulation of the reagent layer 4 is shown in Table 7, the prepared reagent layer coating liquid is heated and stirred for 20min at 40 ℃, and the mixture is uniformly coated on a PET film of a light-transmitting layer, and the wet film thickness is 300 mu m. The diffusion layer 3 was then applied after it had been completely dried in a forced air drying oven at 37℃and a wet film thickness of 750. Mu.m. The diffusion layer formulation is shown in table 8. The reagent sheet was cut into 1cm ² pieces after drying in a fume hood, and the upper frame 1 and the lower frame 7 of the reagent sheet were assembled using a plastic sheet. The diameter of the central opening of the upper and lower outer frames is 10mm.

TABLE 7 reagent layer reagent

Sequence number	Reagent layer reagent	Dosage of
			1	Photographic gelatin	20g/m2
2	Azo arsine III	26mmol/m2
			3	SURFACTANT 10G	6g/m²
4	TritonX-100	1.605g/m²
			5	Buffer solution (MES buffer system)	pH=5.6,0.012mol/m2

TABLE 8 diffusion layer reagent

Example 17

The embodiment provides a dry chemical reagent tablet for quantitatively determining creatinine and a preparation method thereof. The dry chemical reagent sheet structure shown in fig. 1 is adopted, and the specific operation steps are as follows:

The light-transmitting layer 5 is made of PET film, the thickness is 125 μm, and the light transmittance at the wavelength of 540nm is more than 95%. The formulation of the reagent layer 4 is shown in Table 9, and the prepared reagent layer coating liquid was uniformly coated on the PET film as a light-transmitting layer, and the wet film thickness was 300. Mu.m. The diffusion layer 3 was then applied after it had been completely dried in a forced air drying oven at 37℃and the wet film thickness was 1000. Mu.m. The diffusion layer formulation is shown in table 10. The reagent sheet was cut into 1cm ² pieces after drying in a fume hood, and the upper frame 1 and the lower frame 7 of the reagent sheet were assembled using a plastic sheet. The diameter of the central opening of the upper and lower outer frames is 7mm.

Table 9 reagent layer formulation

Table 10 diffusion layer formulation

Example 18

The embodiment provides a dry chemical reagent tablet for quantitatively determining uric acid and a preparation method thereof. The dry chemical reagent sheet structure shown in fig. 1 is adopted, and the specific operation steps are as follows:

The light-transmitting layer 5 is made of PET film, the thickness is 125 μm, and the light transmittance at the wavelength of 670nm is more than 95%. The formulation of the reagent layer 4 is shown in Table 11, and the prepared reagent layer coating liquid was uniformly coated on the PET film as a light-transmitting layer to form a wet film with a thickness of 300. Mu.m. The diffusion layer 3 was then applied after it had been completely dried in a forced air drying oven at 37℃and a wet film thickness of 500. Mu.m. The diffusion layer formulation is shown in table 12. The reagent sheet was cut into 1cm ² pieces after drying in a fume hood, and the upper frame 1 and the lower frame 7 of the reagent sheet were assembled using a plastic sheet. The diameter of the central opening of the upper and lower outer frames is 10mm.

Table 11 reagent layer formulation

Table 12 diffusion layer formulation

Example 19

The embodiment provides a dry chemical reagent tablet for quantitatively determining glucose and a preparation method thereof. The dry chemical reagent sheet structure shown in fig. 1 is adopted, and the specific operation steps are as follows:

The light-transmitting layer 5 is made of PET film, the thickness is 125 μm, and the light transmittance at the wavelength of 540nm is more than 95%. The formulation of the reagent layer 4 is shown in Table 13, and the prepared reagent layer coating liquid was uniformly coated on the PET film as a light-transmitting layer to form a wet film having a thickness of 300. Mu.m. The diffusion layer 3 was then applied after it had been completely dried in a forced air drying oven at 37℃and a wet film thickness of 500. Mu.m. The diffusion layer formulation is shown in table 14. The reagent sheet was cut into 1cm ² pieces after drying in a fume hood, and the upper frame 1 and the lower frame 7 of the reagent sheet were assembled using a plastic sheet. The diameter of the central opening of the upper and lower outer frames is 10mm.

TABLE 13 reagent layer formulation

Table 14 diffusion layer formulation

Example 20

The embodiment provides a dry chemical reagent tablet for quantitatively determining cholesterol and a preparation method thereof. The dry chemical reagent sheet structure shown in fig. 1 is adopted, and the specific operation steps are as follows:

The light-transmitting layer 5 is made of PET film, the thickness is 125 μm, and the light transmittance at the wavelength of 540nm is more than 95%. The formulation of the reagent layer 4 is shown in Table 15, and the prepared reagent layer coating liquid was uniformly coated on the PET film as a light-transmitting layer, and the wet film thickness was 200. Mu.m. The diffusion layer 3 was then applied after it had been completely dried in a forced air drying oven at 37℃and a wet film thickness of 500. Mu.m. The diffusion layer formulation is shown in table 16. The reagent sheet was cut into 1cm ² pieces after drying in a fume hood, and the upper frame 1 and the lower frame 7 of the reagent sheet were assembled using a plastic sheet. The diameter of the central opening of the upper and lower outer frames is 7mm.

TABLE 15 reagent layer formulation

Sequence number	Reagent layer reagent	Dosage of
			1	Photographic gelatin	110g/m²
2	Triton (TritonX-100)	0.67g/m²
			3	Buffer solution (phosphate buffer system)	pH=6.5,13.3g/m²

TABLE 16 diffusion layer formulation

Test example 1

The dry chemical reagent tablets prepared in examples 1 to 9 and comparative examples 1 to 2 were tested, and 5 serum samples having triglyceride concentrations of 0.40mmol/L, 1.385mmol/L, 2.37mmol/L, 4.338mmol/L, and 5.65mmol/L, respectively, were prepared (wherein the serum samples having concentrations of 0.40mmol/L, 2.37mmol/L, and 5.65mmol/L were commercially available composite calibrators, and the remaining concentrations were diluted concentrations of the commercially available composite calibrators). And 5.5 mu L of a sample to be detected is dripped into the sample adding hole 2 of the upper outer frame, the sample uniformly and rapidly permeates onto the reagent layer through the diffusion layer, the reaction is carried out for 5min at 37 ℃, and the optical density value of the color development circular spot is measured by an optical densitometer at the wavelength of 540 nm. The test results of the examples and comparative examples are shown in table 17.

Commercially available composite calibrators having concentrations of 0.40mmol/L, 2.37mmol/L and 5.65mmol/L were added dropwise to the reaction mixture of the examples and comparative examples, and the coefficient of variation CV was calculated 10 times repeatedly, and the results are shown in Table 18.

TABLE 17 Performance parameters of examples 1-9 (S1-S9) and comparative examples 1-2 (D1-D2)

TABLE 18 coefficient of variation CV for examples 1-9 (S1-S9) and comparative examples 1-2 (D1-D2)

As can be seen from tables 17-18, the slope of example 1 was 0.0846, which is higher than that of comparative examples 1-2, indicating a large difference in optical density between high and low concentrations, and the color difference was significant, and the correlation coefficient r ² of example 1 was 0.9993, which is higher than that of comparative examples 1-2, indicating a higher correlation coefficient of triglyceride concentration and optical density. The coefficient of variation CV for example 1 was lower than for comparative examples 1-2, indicating that example 1 was more accurate.

Test example 2

The dry chemical reagent tablets prepared in examples 10 to 14 and comparative examples 3 to 4 were tested, 4 small tablets (each with a size of 1.5cm x 1.5 cm) were taken and the albumin quantitative detection dry tablets prepared in the invention were batched, serum samples with albumin standard concentrations of 13g/L, 20g/L, 35g/L, 46g/L and 62g/L (wherein the serum samples with concentrations of 13g/L, 35g/L and 62g/L are commercial composite calibrators, and the rest concentrations are dilution concentrations of the commercial composite calibrators) were respectively dropped, and after waiting for 5min, optical density values were tested at a wavelength of 630nm by an optical densitometer, wherein the corresponding optical density values also show a gradient change relationship. The test results of the examples and comparative examples are shown in table 19.

Commercially available composite calibrators having concentrations of 13g/L, 35g/L and 62g/L were added dropwise to the reaction mixture of the examples and comparative examples, and the coefficient of variation CV was calculated 10 times repeatedly, and the results are shown in Table 20.

TABLE 19 Performance parameters for examples 10-14 (S10-S14) and comparative examples 3-4 (D3-D4)

TABLE 20 coefficient of variation CV for examples 10-14 (S10-S14) and comparative examples 3-4 (D3-D4)

As can be seen from tables 19 and 20, the correlation coefficient r2 of example 10 is 0.9868, which shows that the optical density and the albumin concentration show a good linear relationship, the slope of example 10 is 0.011, which shows that the difference between the optical densities of high concentration and low concentration is large, and the color difference is obvious, compared with comparative examples 3 to 4. And the coefficient of variation CV value of the embodiment 10 is lower than that of the comparative examples 3-4, the repeatability meets the requirement, and the precision is better.

Test example 3:

the dry reagent tablets prepared in example 15 were 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 (wherein the serum samples with concentrations of 0.04U/mL, 0.70U/mL, and 1.65U/mL were commercial composite calibrators, and the rest were dilution concentrations of the commercial composite calibrators) were prepared. And (3) dropwise adding 11 mu L of a sample to be detected into the sample adding hole 2 of the upper outer frame, uniformly and rapidly penetrating the sample into the reagent layer through the diffusion layer, reacting for 5min at 37 ℃, and measuring the optical density value of the color development circular spot at 670nm by using an optical densitometer. The test results are shown in Table 21.

Commercially available composite calibrators having concentrations of 0.04U/mL, 0.70U/mL and 1.65U/mL were added dropwise to the reaction mixture of the examples and comparative examples, and the coefficient of variation CV was calculated 10 times, and the results are shown in Table 22.

TABLE 21 example 15 (S15) Performance parameters

TABLE 22 coefficient of variation CV for example 15 (S15)

Test example 4

The dry chemical reagent tablets prepared in example 16 were tested, and five serum samples having calcium ion concentrations of 1.8mg/dL, 4.8mg/dL, 9.3mg/dL, 12.45mg/dL, and 13.8mg/dL, respectively, were prepared (wherein the serum samples having concentrations of 1.8mg/dL, 9.3mg/dL, and 13.8mg/dL were commercially available composite calibrators, and the remaining concentrations were diluted concentrations according to the commercially available composite calibrators). And (3) dropwise adding 10 mu L of a sample to be detected into the sample adding hole 2 of the upper outer frame, uniformly and quickly penetrating the sample into the reagent layer through the diffusion layer, reacting for 5min at 37 ℃, and measuring the optical density value of the color development circular spot at 680nm wavelength by using an optical densitometer. The test results of examples and comparative examples are shown in table 23.

Commercially available composite calibrators having concentrations of 1.8mg/dL, 9.3mg/dL and 13.8mg/dL were added dropwise to the reaction mixture of the examples and comparative examples, and the coefficient of variation CV was calculated 10 times repeatedly, and the results are shown in Table 24.

TABLE 23 Performance parameters for example 16 (S16)

TABLE 24 Performance parameters for example 16 (S16)

Test example 5

The dry reagent tablets prepared in example 17 were tested to prepare 5 serum samples having creatinine concentrations of 0.5mg/dL, 0.8mg/dL, 1.5mg/dL, 8.5mg/dL, and 13.2mg/dL, respectively (wherein the serum samples having concentrations of 0.5mg/dL, 1.5mg/dL, and 13.2mg/dL were commercially available composite calibrators, and the remaining concentrations were diluted according to the commercially available composite calibrators). And 5.5 mu L of a sample to be detected is dripped into the sample adding hole 2 of the upper outer frame, the sample uniformly and rapidly permeates onto the reagent layer through the diffusion layer, the reaction is carried out for 5min at 37 ℃, and the optical density value of the color development circular spot is measured by an optical densitometer at the wavelength of 540 nm. The test results of examples and comparative examples are shown in table 25.

Commercially available composite calibrators having concentrations of 0.5mg/dL, 1.5mg/dL and 13.2mg/dL were added dropwise to the reaction mixture of the examples and comparative examples, and the coefficient of variation CV was calculated 10 times repeatedly, and the results are shown in Table 26.

TABLE 25 Performance parameters for example 17 (S17)

TABLE 26 Performance parameters of example 17 (S17)

Test example 6

The dry reagent sheet prepared in example 18 was tested, and 5 serum samples having uric acid concentrations of 0.042mmol/L, 0.263mmol/L, 0.357mmol/L, 0.607mmol/L, and 0.981mmol/L (wherein the serum samples having concentrations of 0.042mmol/L, 0.357mmol/L, and 0.981mmol/L were commercially available composite calibrators, and the remaining concentrations were diluted concentrations of the commercially available composite calibrators), respectively, were prepared. And (3) dropwise adding 10 mu L of a sample to be detected into the sample adding hole 2 of the upper outer frame, uniformly and rapidly penetrating the sample into the reagent layer through the diffusion layer, reacting for 5min at 37 ℃, and measuring the optical density value of the color development circular spot at 670nm by using an optical densitometer. The test results of the examples and comparative examples are shown in table 3.

Commercially available composite calibrators having concentrations of 0.042mmol/L, 0.357mmol/L and 0.981mmol/L were added dropwise to the reaction mixture of examples and comparative examples, and the coefficient of variation CV was calculated 10 times repeatedly, and the results are shown in Table 28.

TABLE 27 example 18 (S18) Performance parameters

TABLE 28 coefficient of variation CV for example 18 (S18)

Test example 7

The dry reagent tablets prepared in example 19 were tested to prepare five serum samples having glucose concentrations of 35mg/dL, 221mg/dL, 300mg/dL, 480mg/dL and 600mg/dL, respectively (wherein the serum samples having concentrations of 35mg/dL, 300mg/dL and 600mg/dL were commercially available composite calibrators, and the remaining concentrations were diluted concentrations of the commercially available composite calibrators). And (3) dropwise adding 10 mu L of a sample to be detected into the sample adding hole 2 of the upper outer frame, uniformly and quickly penetrating the sample into the reagent layer through the diffusion layer, reacting for 5min at 37 ℃, and measuring the optical density value of the color development circular spot at the wavelength of 540nm by using an optical densitometer. The test results are shown in Table 29.

Commercially available composite calibrators were added dropwise to the reaction of the examples and comparative examples at concentrations of 35mg/dL, 300mg/dL and 600mg/dL, respectively, and the coefficient of variation CV was calculated 10 times repeatedly, and the results are shown in Table 30.

TABLE 29 example 19 (S19) Performance parameters

TABLE 30 coefficient of variation CV for example 19 (S19)

Test example 8

The dry reagent tablets prepared in example 20 were tested. 5 serum samples with cholesterol concentrations of 1.80g/L, 2.70g/L, 3.15g/L, 3.60g/L and 4.50g/L were prepared (wherein the serum samples with concentrations of 1.80g/L and 4.50g/L are commercial composite calibrators, and the rest concentrations are dilution concentrations of the commercial composite calibrators). And 5.5 mu L of a sample to be detected is dripped into the sample adding hole 2 of the upper outer frame, the sample uniformly and rapidly permeates onto the reagent layer through the diffusion layer, the reaction is carried out for 5min at 37 ℃, and the optical density value of the color development circular spot is measured by an optical densitometer at the wavelength of 540 nm. The test results are shown in Table 31.

Commercially available composite calibrators having a concentration of 1.80g/L and 4.50g/L were added dropwise to the reaction mixture of the examples and comparative examples, and the coefficient of variation CV was calculated 10 times repeatedly, and the results are shown in Table 32.

TABLE 31 example 20 (S20) Performance parameters

TABLE 32 coefficient of variation CV for example 20 (S20)

It should be noted that the above-described embodiments are only for explaining the present invention and do not constitute any limitation of the present invention. The invention has been described with reference to exemplary embodiments, but it is understood that the words which have been used are words of description and illustration, rather than words of limitation. Modifications may be made to the invention as defined in the appended claims, and the invention may be modified without departing from the scope and spirit of the invention. Although the invention is described herein with reference to particular means, materials and embodiments, the invention is not intended to be limited to the particulars disclosed herein, as the invention extends to all other means and applications which perform the same function.

Claims

1. The dry chemical reagent tablet comprises an upper outer frame, a diffusion layer, a reagent layer, a light-transmitting layer and a lower outer frame from top to bottom in sequence, wherein the upper outer frame is provided with a sample adding hole, and the lower outer frame is provided with a light-transmitting hole;

The quantity ratio of the first particles to the second particles is 1 (0.25-4), preferably 1 (0.5-1.5);

the ratio D2/d1=0.2-0.8 of the particle size D2 of the second particulate matter to the particle size D1 of the first particulate matter, preferably, D2/d1=0.5-0.7.

2. The dry reagent tablet of claim 1, wherein the first particulate matter is the same as or different from the second particulate matter, each independently selected from at least one of titanium dioxide, barium sulfate, diatomaceous earth, kaolin, barium titanate, polystyrene microspheres;

And/or, the adhesive comprises polyisobutylene;

And/or the hydrophilic high molecular polymer comprises at least one of cellulose acetate and cellulose nitrate, preferably, the viscosity of the hydrophilic high molecular polymer is 30-45 poise, preferably 35-40 poise.

3. The dry reagent tablet according to claim 1 or 2, wherein the amount of particulate matter in the diffusion layer reagent is 10-5000 g/m ², preferably 100-1000 g/m ²;

And/or, the dosage of the adhesive in the diffusion layer reagent is 12.5-75 g/m ², preferably 37.5-62.5 g/m ²;

And/or the dosage of the hydrophilic high molecular polymer in the diffusion layer reagent is 1-200 g/m ², preferably 10-100 g/m ².

4. A dry chemical reagent sheet according to any one of claims 1 to 3, wherein the particle size of the first particulate matter in the diffusion layer reagent is 4 to 8 μm, preferably 4.5 to 5.5 μm;

and/or the second particulate matter has a particle size of 1 to 4 μm, preferably 2.5 to 3.5 μm.

5. A dry chemical reagent sheet according to any one of claims 1 to 3, wherein the particle size of the first particulate matter in the diffusion layer reagent is 40 to 80 μm, preferably 45 to 55 μm;

and/or the particle size of the second particles is 10 to 40 μm, preferably 25 to 35 μm.

6. The dry chemical reagent tablet according to any one of claims 1 to 5, wherein the diffusion layer reagent further comprises a surfactant,

Preferably, the surfactant comprises at least one of tween, triton, surfynol, S24 and fluorine surfactant,

7. The dry chemical reagent tablet according to any one of claims 1 to 6, wherein the reagent layer reagent comprises at least one of a buffer system, a biological enzyme, a color developer, a binder, a surfactant, a colloid, and an alkaline agent.

8. The dry chemical reagent tablet according to any one of claims 1 to 7, wherein the wet film thickness of the reagent layer is 100 to 600 μm and the dry film thickness is 10 to 100 μm;

and/or the wet film thickness of the diffusion layer is 300-1600 mu m, and the dry film thickness is 200-800 mu m;

and/or the thickness of the light-transmitting layer is 20-400 μm.

9. A method of preparing a dry chemical reagent tablet according to any one of claims 1 to 8, comprising the steps of:

step 2, coating a diffusion layer reagent on the reagent layer, and drying;

And 3, assembling the upper outer frame and the lower outer frame.

10. The use of a dry chemical reagent strip according to any one of claims 1-8 for quantitative detection of a substance to be detected 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;