CN110791546A

CN110791546A - Dry tablet reagent for quantitative detection of urea nitrogen

Info

Publication number: CN110791546A
Application number: CN201911055190.6A
Authority: CN
Inventors: 郑日记; 黄利寒; 甘斌; 武伟民; 李菁杨
Original assignee: Wuxi Jinbucheng Medical Equipment Technology Co Ltd
Current assignee: Wuxi Jinbucheng Medical Equipment Technology Co Ltd
Priority date: 2019-10-31
Filing date: 2019-10-31
Publication date: 2020-02-14

Abstract

The invention provides a dry sheet reagent for quantitatively detecting urea nitrogen, which comprises a shell, wherein a diffusion layer, a reagent layer, a resin layer and a base material are sequentially attached to the shell, a sample dripping hole is formed in the center of the upper surface of the shell, a light transmission hole is formed in the center of the bottom of the shell, the diffusion layer, the reagent layer and the resin are sequentially attached to the base material in a layering mode from top to bottom through a coating process, the diffusion layer mainly comprises cellulose acetate, titanium dioxide, a surfactant, magnesium chloride, glycerol and polyethylene glycol, and the reagent layer mainly comprises urease, phenol red sodium salt, a phosphate buffer solution, polyacrylamide, polyvinylpyrrolidone, gelatin and an enzyme protective agent EDTA. The invention solves the problems of unstable wet chemical detection reagent and larger error of test result in the prior art.

Description

Dry tablet reagent for quantitative detection of urea nitrogen

Technical Field

The invention relates to a urea nitrogen detection reagent, in particular to a dry sheet reagent for quantitative detection of urea nitrogen.

Background

So-called "dry-patch reagents" are relatively conventional "wet chemistry". The method is a mode that liquid in a detected sample is used as a reaction medium, and an object to be detected directly reacts with a reagent fixed on a carrier, and the method is different from the traditional wet chemistry in the medium participating in the chemical reaction. With the development of techniques for separation, purification, storage, etc. of enzymes in biochemistry, the progress of sensor, photometer, and electrode techniques, and the popularization of computers, dry chemistry has also rapidly developed.

Dry chemistry has unique advantages over wet chemistry, and as for reagents, dry chemistry reagents do not need preparation before measurement, do not need calibration, and are stable, and whole blood can be directly measured; as for the instrument, a water supply and drainage system and a pure water system are not required, so that the operation is simple and the maintenance is easy.

Disclosure of Invention

The invention provides a dry sheet reagent for urea nitrogen quantitative detection, which solves the problems of unstable wet chemical detection reagent and larger error of test results in the prior art.

In order to solve the technical problem, the invention provides a dry plate reagent for quantitative detection of urea nitrogen, which comprises a shell, the diffusion layer, the reagent layer, the resin layer and the base material are sequentially adhered in the shell, a sample dripping hole is formed in the center of the upper surface of the shell, a light transmitting hole is formed in the center of the bottom of the shell, the diffusion layer, the reagent layer and the resin are sequentially laminated on the base material from top to bottom through a coating process, the diffusion layer mainly comprises cellulose acetate, titanium dioxide, a surfactant, magnesium chloride, glycerol and polyethylene glycol, the reagent layer mainly comprises urease, pigment, buffer solution, cross-linking agent, gel and enzyme protective agent, the dye is phenol red sodium salt, the buffer solution is phosphoric acid buffer solution, the cross-linking agent is polyacrylamide or polyvinylpyrrolidone, the gel is gelatin, and the enzyme protective agent is EDTA.

The resin layer is a water-based epoxy resin.

The substrate is a pet material for providing a skeleton for supporting the reagent, the substrate having a transparency greater than 80%.

The shell is made of plastic and formed by ultrasonic bonding of upper and lower layers of plastic in a pressing mode, and is used for protecting the reagent from deformation caused by various external forces in the processes of transportation, storage and testing, so that the reagent is prevented from being damaged.

The structure of the diffusion layer takes cellulose acetate as a three-dimensional network framework, and the three-dimensional network framework is wrapped with a corresponding reagent.

The reagent layer is formed by wrapping corresponding reagents by taking coagulated and dried gelatin as a carrier.

The invention has the following beneficial effects: the dry tablet reagent is established on the basis of wet chemistry and used for a dry chemistry method for testing urea nitrogen, has excellent performance, long storage time and accurate test result, can obviously improve various performance indexes of uric acid in the detection and storage processes, and makes up for the deficiency of wet chemistry.

Drawings

FIG. 1 is a schematic cross-sectional view of the dry plate reagent for quantitative detection of urea nitrogen in accordance with the present invention.

FIG. 2 is a schematic diagram of the overall structure of the dry tablet reagent of the present invention.

Wherein, 1-sample, 2-diffusion layer, 3-reagent layer, 4-resin layer, 5-base material, 6-shell, 7-sample dropping hole and 8-light hole.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail below with reference to the accompanying drawings and specific embodiments.

As shown in figure 1, the invention provides a dry sheet reagent for quantitative detection of urea nitrogen, which comprises a shell 6, wherein a diffusion layer 2, a reagent layer 3, a resin layer 4 and a substrate 5 are sequentially attached in the shell, a sample dripping hole 7 is formed in the center of the upper surface of the shell 6, and a light hole 8 is formed in the center of the bottom of the shell. The diffusion layer 2, the reagent layer 3 and the resin 4 are laminated on the base material 5 sequentially from top to bottom through a coating process.

Diffusion layer 2, diffusion layer are with cellulose acetate as three-dimensional network skeleton, are wrapping up corresponding reagents such as titanium dioxide in the similar sponge of structure, and the main function is including evenly diffusing the sample, macromolecular material and interference material in the filtration sample, and the diffusion layer mainly comprises cellulose acetate, titanium dioxide, surfactant active, magnesium chloride, glycerine, polyethylene glycol etc.. The content of each diffusion layer component in each dry sheet is shown in table 1:

table 1: content of each component of diffusion layer

Name of reagent	Content per tablet (unit g)
		Cellulose acetate	0.0025
Titanium dioxide	0.03
		Surface active agent	0.00023
Magnesium chloride	0.00122
		Glycerol	0.0005
Polyethylene glycol	0.0005

The reagent layer 3 mainly comprises urease, phenol red sodium salt, phosphate buffer, polyacrylamide, polyvinylpyrrolidone, gelatin, enzyme protective agent EDTA and other necessary biochemical reagents, and the reagent layer takes coagulated and dried gelatin as a carrier to wrap the corresponding reagent, and has a structure similar to jelly. The contents of the components of the reagent layer in each dry tablet are shown in table 2:

table 2: the content of each component of the reagent layer

Name of reagent	Each tablet content
		Urease	1.4U
Phenol Red sodium salt	0.000019G
		Phosphate buffer	25UL
Polyacrylamide	0.03667G
		Polyvinylpyrrolidone	0.0453G
Gelatin	0.083G
		EDTA	0.01667G
Triton-100	0.00064G

The resin layer 4 can effectively attach the reagent layer to the base material, can reduce the requirements on the material and performance of the base material, can use the base material which is not treated, and avoids the interference of the treated base material to the detection reagent, and the resin layer is water-based epoxy resin and has a structure similar to common liquid glue. The resin material is purchased and directly used by the original factory.

A substrate 5, which mainly provides a skeleton supporting the reagents, the transparency of which must be greater than 80%, the substrate being pet material, structured like a hard plastic paper sheet.

The shell 6, this shell layer is mainly that the protection reagent is in the transportation, stores, and the deformation that the in-process of test leads to because various power to avoid the damage of reagent, especially in last quick-witted test, no matter be automatic or semi-automatic send into the dry film reagent detection machine, the dry film inevitably receives corresponding power, the damage of the dry film of reagent can be avoided to the reagent shell, the shell is formed by upper and lower layer plastics pressfitting ultrasonic bonding, the overall structure of dry film reagent is as shown in figure 2.

The preparation method of the dry tablet reagent comprises the following steps:

1. a resin material is prepared to be coated on a base material.

2. The resin is coated with a reagent, and the reagent layer is coated by different coating methods according to needs and processes. The thickness and the roughness requirement on reagent layer are strict, and for guaranteeing the poor and stability between the batches of reagent, the thickness on reagent layer need strict control error, and wet thickness error control is in 300 +/-10 um, and dry thickness error need be controlled in 60 +/-3 um.

3. A diffusion layer is coated on the reagent. The coating thickness of the diffusion layer is 300 +/-20 um.

In summary, the reagent is coated on the transparent plastic film in the form of a film to be made into a dry sheet, and the dry sheet reagent corresponding to the concentration of the target substance is obtained by detecting the reflected light. The dry chemical reagent prepared by the method has excellent performance and long storage time.

The specific operation of the dry-film reagent detection of the invention is as follows:

as shown in figure 1, 10ul of sample 1 is added on the surface of the diffusion layer, the sample 1 can be transversely and longitudinally uniformly diffused downwards through the diffusion layer and enters the reagent layer, target substance urea nitrogen in the sample is catalyzed by urease to generate NH3, and NH3 reacts with yellow phenol red sodium salt to generate a mauve substance. After sample adding, light with the wavelength of 540nm is used for irradiating the position of the center of the base material, the change of the reflected light of the dry plate after sample adding is recorded, the density of the reflected light is in inverse proportion to the concentration of the target substance urea nitrogen, and the corresponding concentration, namely the detection concentration of the sample corresponding to uric acid, is calculated through a computer. The equation for the reaction of the target substance urea nitrogen with the corresponding substance in the dry tablet reagent is as follows:

in conclusion, the invention provides the dry tablet reagent which is established on the basis of wet chemistry and is used for the dry chemistry method for testing urea nitrogen, the dry tablet reagent has excellent performance, long storage time and accurate test result, can obviously improve each performance index of uric acid in the detection and storage processes, and makes up the deficiency of wet chemistry.

The above description is only an example of the present invention, and is not intended to limit the present invention, and it is obvious to those skilled in the art that various modifications and variations can be made in the present invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the claims of the present invention.

Claims

1. The dry sheet reagent for quantitative detection of urea nitrogen is characterized by comprising a shell (6), wherein a diffusion layer (2), a reagent layer (3), a resin layer (4) and a substrate (5) are sequentially attached to the shell, a sample dripping hole (7) is formed in the center of the upper surface of the shell (6), a light hole (8) is formed in the center of the bottom of the shell, the diffusion layer (2), the reagent layer (3) and the resin (4) are sequentially attached to the substrate (5) in a layering manner from top to bottom through a coating process, the diffusion layer (2) mainly comprises cellulose acetate, titanium dioxide, a surfactant, magnesium chloride, glycerol and polyethylene glycol, the reagent layer (3) mainly comprises urease, a pigment, a buffer solution, a cross-linking agent, a gel and an enzyme protective agent, wherein the pigment is phenol red sodium salt, and the buffer solution is a phosphate buffer solution, The cross-linking agent is polyacrylamide or polyvinylpyrrolidone, the gel is gelatin, and the enzyme protective agent is EDTA.

2. The dry sheet reagent for quantitative detection of urea nitrogen as claimed in claim 1, wherein the resin layer (4) is an aqueous epoxy resin.

3. The dry plate reagent for quantitative detection of urea nitrogen as claimed in claim 1, wherein the substrate (5) is pet material for providing a skeleton for supporting reagent, and the transparency of the substrate is more than 80%.

4. The dry plate reagent for quantitative urea nitrogen detection as claimed in claim 1, wherein the housing (6) is made of plastic, and is formed by laminating and ultrasonically bonding an upper layer and a lower layer of plastic, and is used for protecting the reagent from deformation caused by various external forces during transportation, storage and test, so as to avoid damage to the reagent.

5. The dry plate reagent for quantitative urea nitrogen detection as claimed in claim 1, wherein the structure of the diffusion layer (2) is cellulose acetate as a three-dimensional network skeleton, and the three-dimensional network skeleton is wrapped with the corresponding reagent.

6. The dry plate reagent for quantitative detection of urea nitrogen as claimed in claim 1, wherein the reagent layer (3) is formed by coating corresponding reagent with coagulated and dried gelatin as carrier.