JPH054030B2 - - Google Patents

Description

[Detailed description of the invention]

[Industrial Field of Application] The present invention relates to a dry analytical element containing at least one layer of a synthetic binder, and more particularly to a dry-operable integrated multilayer analytical element for the quantitative analysis of proteins such as albumin or total protein in aqueous liquids. It is related to. [Prior Art] In dry integrated multilayer analytical elements used for quantitative analysis of proteins such as albumin and total protein in aqueous liquids, non-containing materials are used to stabilize reagents and avoid competitive reactions with other proteins. As a proteinaceous hydrophilic polymer binder, polyacrylamide, poly-N-vinyl-2-pyrrolidone, acrylamide-N-vinyl-2-pyrrolidone copolymer or agarose is used as disclosed in JP-A-58-77664 and JP-A-58-77- This is proposed in Publication No. 50660. These elements have a reagent layer in which a reagent and a buffer agent are dispersed in the above-mentioned non-protein hydrophilic polymer binder and a porous spreading layer coated on a polyethylene terephthalate support in this order. When human serum is spotted on the development layer, moisture is supplied from the development layer to the reagent layer, and both layers come into liquid contact. Reagents are supplied from the reagent layer and diffused into the development layer, and the development layer releases albumin or total serum. Since the protein reacts with the reagent to form a color, it is an element of a dry operation in which the color density is measured and the content of each is determined using the principle of colorimetric measurement. [Problems to be solved by the invention] However, since the non-protein hydrophilic polymer binder used in these elements is usually difficult to cure, the quality of the dry analytical elements produced, especially the poor film quality, is a problem. It was hot. Furthermore, in the albumin analysis element, it was found that as the measurement time was prolonged, globulin interference appeared to a non-negligible extent. An object of the present invention is to provide a dry analysis element in which a non-proteinaceous hydrophilic polymer binder can be crosslinked and cured, the film quality is strengthened by curing, and the element can be stably manufactured. Another object of the present invention is to provide a dry analytical element for albumin analysis that has a simple layer structure and reduces errors due to interference by globulin and other proteins. Another object of the present invention is to provide a dry analytical element for total protein analysis whose film quality is reinforced by the use of a curable non-proteinaceous hydrophilic polymer binder. [Means for Solving the Problems] This object of the present invention is to produce a copolymer of methallyl alcohol and acrylamide or a derivative thereof, acrylic acid or a derivative thereof, or N-vinyl-2-pyrrolidone in a non-proteinaceous manner. This is achieved by means of a dry integrated multilayer analytical element containing in at least one layer a hydrophilic polymer binder. Methallyl alcohol is also called 2-methylallyl alcohol, and is thought to have the following structure as a polymerized unit (constituent repeating unit). Comonomers copolymerized with methallyl alcohol include acrylamide and its derivatives (for example, methacrylamide, N,N-dimethylacrylamide, diacetone acrylamide, N-(tert
-butyl)acrylamide, N-(isopropyl)
acrylamide, 2-acrylamido-2-methylpropanesulfonic acid, N-[3-(dimethylamino)propyl]acrylamide, etc.), acrylic acid and its derivatives (for example, hydroxyethyl acrylate, etc.). -Vinyl-2-pyrrolidone and acrylic acid are preferred, and acrylamide and N-vinyl-2-pyrrolidone are particularly preferred. Two or more of these comonomers can also be used in combination. The copolymerization ratio is 0.5 to 50 methallyl alcohol units.
It is about mol %, preferably about 3 to 30 mol %, and therefore, the other vinyl compound units such as acrylamide are about 50 to 99.5 mol %, preferably about 70 to 97 mol %. These copolymers containing methallyl alcohol are produced using the “Polymer Synthesis Method” (edited by CG Overberger, translated by Hiroshi Minato,
It can be easily synthesized by selecting an appropriate catalyst and reaction solvent with reference to "Experimental Methods of Polymer Synthesis" (co-authored by Furuyuki Otsu and Masayoshi Kinoshita, published by Kagaku Doujin, 1972) (published by Tokyo Kagaku Doujin, 1969). be able to. Among the copolymers, acrylamide methallyl alcohol copolymer, acrylamide N-vinyl-2-pyrrolidone-methallyl alcohol copolymer, and acrylamide acrylic acid-methallyl alcohol copolymer are preferred. The synthesized copolymer can be easily crosslinked and cured by adding a crosslinking agent. Crosslinking agents include formalin, aldehydes such as glutaraldehyde, epoxy compounds containing at least two or more glycidyl groups such as tetraethylene glycol and ethylene glycol diglycidyl ether, and divinyl sulfone, ethylene bis vinyl sulfonylacetamide, and bis(vinyl A compound containing at least two electron-withdrawing vinyl groups such as sulfonylmethyl)ether can be used.The amount of the crosslinking agent added is 1.25% by weight or more, preferably 2 to 20% by weight, based on the methallyl copolymer. Selected from a range of %. Crosslinking is generally carried out by heating, and the temperature varies depending on the composition of the copolymer and the type of crosslinking agent, and crosslinking can be performed at a temperature range of about 20°C to about 200°C, but for dry analysis elements, the temperature is about 20°C. to about 80℃, preferably about
Selected from a temperature range of 30°C to approximately 60°C. The dry analytical element of the present invention usually comprises at least one hydrophilic polymer binder layer on a light-transparent water-impermeable support and a porous spreading layer provided thereon to uniformly diffuse the sample. It's summery. Typical examples of such multilayer analytical elements for measuring aqueous liquids are U.S. Pat.
It is disclosed in No. 4042335. In some cases, the element may consist of a single zone, as in US Pat. No. 3,485,587. Examples of the porous spreading layer include woven fabric spreading layers (e.g., plain weave such as broadcloth and poplin) described in JP-A-55-164356 and JP-A-57-66359, etc., and JP-A-60-222769. The knitted fabric development layer described in publications (e.g.
tricot, double tricot, Milanese, etc.), a spreading layer made of organic polymer fiber pulp-containing paper described in JP-A-57-148250, a membrane described in JP-A-53-21677, USP 3,992,158, etc. Filter (brushed polymer layer),
A non-fibrous isotropic porous spread layer such as a porous layer containing continuous micropores in which polymer microbeads, glass microbeads, and diatomaceous earth are held in a hydrophilic polymer binder, the polymer described in JP-A-55-90859. A non-fibrous isotropic porous spread layer consisting of a porous layer containing continuous micropores (three-dimensional lattice-like granular structure, etc.) in which microbeads are bonded in point contact with a polymer adhesive that does not swell with water, and other special features. 1977-
148250, a spreading layer made of paper containing polyethylene pulp, JP-A-56-24576,
The spreading layer made of paper described in JP-A No. 57-94658, the fibrous spreading layer coated with a fiber dispersion described in JP-A-57-125847, etc. can be used. Among these porous spreading layers, fibrous spreading layers, such as woven fabric spreading layers and knitted fabric spreading layers, are preferred because they can easily contain and retain the indicator. By providing such a polyphilic expansion layer, serum,
Aqueous liquid samples such as urine can be uniformly dispersed to improve analysis accuracy and reproducibility. The hydrophilic polymer binder layer is a water-absorbing layer whose main component is an uncrosslinked, partially crosslinked, or crosslinked product of the methallyl alcohol copolymer, and is a layer that can swell upon contact with water and absorb water. Swelling rate when absorbing water is about 150% to about 2000 at 30℃
%, preferably in the range of about 250% to about 1500%. The content of the hydrophilic polymer binder used in the hydrophilic polymer binder layer is from about 7 g to about 70 g, preferably from about 10 g to about 50 g ^per square meter.
is within the range of The hydrophilic polymer binder can also be used in combination with other hydrophilic binders if necessary. The dry thickness of this hydrophilic polymeric binder layer ranges from about 1 μm to about 100 μm, preferably about 3 μm to about 30 μm, and the coverage ranges from about 1 g/m ² to about
100g/m ² , preferably about 3g/m ² to about 30g/m 2
A range of m ² is suitable. This hydrophilic polymer binder layer has a pH value from 2.0 when performing analysis operations using liquid samples.
The buffer may be appropriately selected from known buffering agents capable of buffering to a desired value in the range of 10.0. Buffers that can be used are listed in "Chemical Handbook Basic Edition" edited by the Chemical Society of Japan (Tokyo, Maruzen Co., Ltd., published in 1966) 1312
-1320 pages, “Data for Biochemical Research”, edited by R, M, C, Dawson et al., 2nd edition
Oxford at thc Clarendon Press, published in 1969)
pp. 476-508, "Biochemistry" 5 , pp. 467 et seq. (1966), "Analytical Biochemistry" 104 ,
There is a PH buffer system described in pages 300-310 (1980). As a specific example of a PH buffer in the range of PH2.0 to 10.0,
Buffers containing tris(hydroxymethyl)aminomethane (Tris); Buffers containing phosphate; Buffers containing borate; Buffers containing acetic acid or acetate; Buffers containing chelic acid or citrate ; Buffer containing glycine; malic acid; succinic acid; malonic acid; tartaric acid; glutaric acid; 3,3 dimethylglutaric acid; N,N-bis(2-hydroxyethyl)glycine (Bicine); N-2-hydroxyethyl piperazine-N'-2-hydroxypropane-3-
Sulfonic acid (HEPPS) Na salt or K salt, etc.; N-
2-hydroxyethylpiperazine-N'-3-sulfonic acid (EPPS) Na salt or K salt, etc.; N-[tris(hydroxymethyl)methyl]-3-aminopropanesulfonic acid (TAPS) Na salt or K salt, etc.; N-2-hydroxyethylpiperazine-
N'-2-ethanesulfonic acid (HEPES) Na salt or K salt; and acids, alkalis, or salts optionally combined with any of these. Specific examples of preferred buffers include potassium dihydrogen phosphate-disodium hydrogen phosphate; Tris-sodium borate; Tris-sodium borate-EDTA.2Na
Salt; Tris-sodium phosphate; acetic acid-sodium acetate; tris-sodium phosphate; Bicine;
HEPPS; HEPPS sodium salt; EPPS; EPPS
Sodium salt; TAPS; TAPS sodium salt;
HEPES; malic acid; succinic acid; malonic acid; tartaric acid; glutaric acid; 3,3-dimethylglutaric acid and the like. The content of PH buffer or acid in the element is 1
It ranges from about 30 milliequivalents to about 500 milliequivalents, preferably from about 50 milliequivalents to about 300 milliequivalents per m ² . The hydrophilic polymer binder layer can contain various components that do not adversely affect the analyte or reagent. Examples include nonionic surfactants. Specific examples of nonionic surfactants include p-octylphenoxypolyethoxyethanol, p-nonylphenoxypolyethoxyethanol, polyoxyethylene oleyl ether, polyoxyethylene sorbitan monolaurate, p-nonylphenoxy Examples include polyglycidol and octyl glucoside. By incorporating a nonionic surfactant into the hydrophilic polymer binder layer, water in an aqueous liquid sample can be absorbed substantially uniformly into the hydrophilic polymer binder layer during analysis operations, and the water can be easily absorbed into the hydrophilic polymer binder layer during analysis operations. Liquid contact is rapid and substantially uniform. The hydrophilic polymer binder layer is optionally 2
More than one layer can be provided. When providing two or more hydrophilic polymer binder layers, a layer close to the porous spreading layer contains a low molecular weight PH buffer composition or a low molecular weight acid, and a layer far from the porous spreading layer contains a high molecular weight one. It is preferable to contain a PH buffer or a high molecular weight acid. Examples of high molecular weight acids that can be used include known carboxyl group-containing polymers and sulfonic acid group-containing polymers.
The hydrophilic polymer binder layer can also contain other known mordants, polymer mordants, and the like. The hydrophilic polymer binder layer is preferably substantially transparent, but if necessary, titanium dioxide fine particles, barium sulfate fine particles,
Optical performance can be adjusted by dispersing a small amount of carbon black or the like. The hydrophilic polymer binder layer can contain a reagent composition described below to function as a reagent layer. In this case, a water absorbing layer may be further provided between the hydrophilic polymer binder layer and the support. The water absorption layer is a layer whose main component is a hydrophilic polymer that absorbs water and swells.It is a layer that can absorb water from an aqueous liquid sample that has reached or penetrated the interface of the water absorption layer.When using a whole blood sample, It has the effect of promoting the penetration of plasma, which is an aqueous liquid component, into the reagent layer or the reagent composition-containing spreading layer. The hydrophilic polymer used in the water absorption layer has a swelling rate of 30℃ when water is absorbed.
from about 150% to about 2000%, preferably from about 250% to about 1500%. Specific examples of hydrophilic polymers include non-crosslinked methallyl alcohol copolymers, acid-treated gelatin disclosed in JP-A-59-171864 and JP-A-60-115850, gelatin such as deionized gelatin, and metalized gelatin. Gelatin, gelatin derivatives such as hydroxyacrylate grafted gelatin, JP-A-59-171864, JP-A-60-
Examples include agarose, bullulan, bullulan derivatives, polyacrylamide, polyvinyl alcohol, polyvinylpyrrolidone, and the like, which are disclosed in No. 115859 and the like. These hydrophilic polymers can be used alone or in combination of two or more. The dry thickness of the water absorption layer is approximately 1μm to approximately 100μm,
Preferably in the range of about 3 μm to about 30 μm, with a coverage of about 1 g/m ² to about 100 g/m ² , preferably about 3
g/m ² to about 30 g/m ² . The water absorption layer can contain a known pH buffering agent, organic carboxylic acid, acidic polymer, basic polymer, etc. to adjust the pH during use (during analytical operations). Furthermore, the water-absorbing layer may contain a known mordant, polymer mordant, or the like. The water absorbing layer is preferably substantially transparent, but if necessary, the optical performance can be adjusted by incorporating a small amount of titanium dioxide fine particles, barium sulfate fine particles, carbon black, etc., dispersed in the water absorbing layer. A reagent composition containing at least one enzyme that reacts with a predetermined component in an aqueous liquid sample to produce a detectable change is dispersed in a single layer or as appropriate in a hydrophilic polymer binder layer or a porous spreading layer. and substantially uniformly contained. Detectable changes mainly mean changes that can be detected by optical measurement methods, such as color changes, color development, fluorescence generation, changes in absorption wavelength in the ultraviolet region, and turbidity generation. The reagent composition depends on the biochemical or chemical reaction involving (catalyzing) the analyte in the aqueous liquid sample and the enzyme selected to analyze this component; In the case of reactions involving enzymes, reagent compositions containing those enzymes can be mixed and contained in one reagent layer, or if necessary, the reagent compositions can be divided into two or more separate layers. It can also be contained. Examples of reagent compositions containing at least one enzyme include U.S. Pat.
Improved Trinder reagent composition for glucose analysis containing glucose oxidase and peroxidase as described in JP-A-59-46854, JP-A-59-54962, JP-A-55-25840, etc.
56-45599, a reagent composition for cholesterol analysis containing cholesterol oxidase, peroxidase, and cholesterol esterase optionally blended as described in JP-A-59-193352, etc., JP-A-59-193352, etc.
-3488, urea nitrogen (BUN) analysis reagent composition containing urease described in JP-A-58-77661, JP-A-56-70460, etc., lipoprotein lipase and glycerol described in JP-A-53-24893, etc. Kinase,
Reagent composition for triglyceride or glycerol analysis containing α-glycerol-3-phosphate oxidase and peroxidase, reagent for bilirubin analysis containing bilirubin-specific oxidase and peroxidase described in JP-A-54-151193 and JP-A-60-78580 Composition, JP-A-53-26188, JP-A-59-193352
There are reagent compositions for uric acid analysis containing uricase and peroxidase described in JP-A No. 55-124499 and JP-A No. 58-86457, color reagent compositions for detecting hydrogen peroxide containing peroxidase, etc. In the case of the dry analytical element for albumin analysis, which is an embodiment of the present invention, Korthoff (IM) is used as an indicator that changes color when combined with albumin.
Kolthoff) Acid-Base Indicators
(Mac Millan, published in 1937) 350
It is preferable to include an acid-base indicator dye that shows protein error as described on page 353. Examples of acid-base indicator dyes that indicate protein errors include bromcresol green, bromcresol purple, bromcresol blue, bromphenol blue, chlorphenol red, phenol red, cresol red, thymol blue,
There are sulfonephthalein indicator dyes such as cresolphthalein, indigoid dyes such as indigo carmine, and azo dye indicators such as methyl red and methyl orange. Among these indicators, sulfonephthalein indicator dyes are preferred, with bromcresol green (BCG) and bromcresol purple (BCP) being most preferred. This indicator is usually contained in the porous spreading layer. In that case, the amount of acid-base indicator dye is 1 m ²
The amount ranges from about 0.2 g to about 3.0 g, preferably from about 0.5 g to about 1.5 g. The indicator may be contained in other layers, such as a hydrophilic polymer binder layer. The PH buffering agent lowers the PH value of the porous spread layer in the liquid contact area during analytical operations from about 2.0 to about 4.0, preferably about 3.2.
Use a low molecular weight PH buffer composition or low molecular weight acid that can be maintained in the range of about 3.4 to 3.4. Examples of such PH buffering agents include low molecular weight carboxylic acids and their salts such as malic acid, lactic acid, succinic acid, malonic acid, laconic acid, tartaric acid, etc., USP3438737, "Chemical Handbook Basic Edition" edited by the Chemical Society of Japan. (Tokyo, Maruzen, 1966
Other low-molecular-weight PH buffer compositions can be mentioned, such as those described on pages 1312-1320 (published in 2013). Among these, malic acid is preferred. In the case of a dry analytical element for total protein analysis, which is also an embodiment of the present invention, for example, Biuret's reagent is utilized. That is, water-soluble cupric salts such as copper sulfate and copper nitrate, chelating agents such as tartrates, citrates, ethylenediamine and the like, and alkalis such as lithium hydroxide and calcium hydroxide are contained. An example of a dry analytical element for total protein analysis using Biuret's reagent is published in Japanese Patent Application Laid-Open No.
Publication No. 101398, Japanese Patent Application Publication No. 1987-77664, Japanese Patent Application Publication No. 1987-77664
It is disclosed in Japanese Patent Application No. 59-51356, Japanese Patent Application No. 132057/1980, etc. This Biuret reagent is preferably contained in a hydrophilic polymer binder layer crosslinked with a methallyl alcohol copolymer. When analyzing other substances, a reagent suitable for the substance to be analyzed is contained in a porous development layer or a hydrophilic polymer binder layer. As the light-transmitting/impermeable support, known supports used in conventional integrated multilayer analysis elements can be used. Specific examples thereof include polyethylene terephthalate, polycarbonate of bisphenol A, polystyrene, and polymers of cellulose esters (e.g., cellulose diacetate, cellulose triacetate, cellulose acetate propionate, etc.) with a thickness of about 50 μm to about 1 mm; A smooth, planar support may be used that is transparent, preferably in the range of about 80 μm to about 300 μm, ie transparent to electromagnetic radiation in a wavelength range of at least a portion of the wavelength range of about 200 nm to about 900 nm.
A known subbing layer or adhesive layer can be provided on the surface of the support to strengthen the adhesion with the hydrophilic polymer binder layer or water absorption layer. An integral multilayer analytical element comprising a hydrophilic polymer binder layer of a crosslinked product of such a methallyl alcohol copolymer can be prepared, for example, by first depositing the methallyl alcohol copolymer, the crosslinking agent and the layer on a transparent support. A hydrophilic polymer binder layer is formed by applying a solution containing a reagent, etc., and heat-treating the applied product at a temperature appropriately selected from the curing temperature range. Then, a porous development layer may be laminated thereon. The dry analysis element of the present invention has a side of about 15 mm to about 30 mm.
Cut into small pieces such as squares or circles of approximately the same size.
Publication No. 142454, Japanese Unexamined Patent Application Publication No. 57-63452, Jitsugakusho
58-32350, Japanese Patent Application Laid-open No. 58-501144, etc., and used as a chemical analysis slide for albumin analysis, it is possible to
It is preferable from all viewpoints such as transportation, storage, and measurement operations.
Depending on the purpose of use, it may be used in the form of a long tape and stored in a cassette or magazine, or small pieces may be attached or stored in a card with an opening. The dry analytical element of the present invention can perform analysis in a liquid sample by the operations described in the patent specification and the like mentioned above. For example, a droplet of an aqueous liquid sample such as plasma or serum in the range of 6μ to 15μ is placed on a porous spreading layer, and
Incubation is carried out for a period of 10 minutes, preferably at a temperature around 37°C, and the optical density of the reagent development layer is measured by reflection photometry using light at or near the maximum absorption wavelength of the color developed from the light-transmissive support side. However, the amount of the analyte in the liquid sample can be determined by the principle of colorimetric measurement using a calibration curve prepared in advance. Quantitative analysis can be performed with high precision by keeping the amount of aqueous liquid sample spotted, incubation time, and temperature constant. This measurement operation is
Publication No. 77746, Japanese Patent Application Laid-open No. 1983-21566, Japanese Patent Application Publication No. 1987-21566
The chemical analyzer described in Japanese Patent No. 161867 and the like allows highly accurate measurements with extremely easy operation. [Function] By copolymerizing methallyl alcohol, hydrophilicity can be imparted, and the hydroxyl group can be reacted with a crosslinking agent to cure the material. Comonomers such as acrylamide and N-vinylpyrrolidone also impart hydrophilicity to the copolymer. When a crosslinked product of such a methallyl alcohol copolymer is used in a hydrophilic polymer binder layer, the PH
The hydrophilic polymer binder layer containing the buffer swells and absorbs water upon contact with the PH buffer and water. This hydrophilic polymer binder layer makes the low molecular weight PH buffer porous when water in the aqueous liquid sample spotted on the porous spreading layer during analytical operations reaches this layer and the two layers come into liquid contact. It has a function of supplying the liquid sample to the developing layer by diffusion and maintaining the area where the liquid sample exists in the porous developing layer at a predetermined constant PH value or a value in the vicinity thereof. [Examples] Example 1 Acrylamide/N-vinyl-2-pyrrolidone/methallyl alcohol terpolymer of the present invention (abbreviated as A.NV.M), molar ratio 58:38:4, viscosity
400 cp (20% solution, 40°C, B-type viscometer) was added with malic acid as a reagent, 1% by weight of formalin and a few drops of 6N-hydrochloric acid based on the copolymer as reagents, and a coating solution was prepared using 180 μm transparent polyethylene. It was coated onto a terephthalate (PET) support to a dry thickness of 35 μm. As a comparative example, acrylamide-N-vinyl-2-pyrrolidone copolymer (abbreviated as A/NV, molar ratio 60:
40, viscosity 400 cp [20% solution at 40°C, B-type viscometer]) and applied in the same manner. Next, the thickness is about 140μ
A broad fabric made of spun cotton yarn (100S) containing approximately 0.7 g/m ² of bromcresol green was laminated to form a porous spreading layer. The amount of reagent applied to the sample thus prepared was as follows.

[Table] When the two coatings prepared in this way were immersed in hot water at 70°C, the sample of the present invention did not dissolve, but the comparative sample began to dissolve, indicating a difference in film quality. . Next, human albumin solutions of different concentrations were spotted and the color density was measured at 640 nm. The results showed that the analytical element using the crosslinkable and curable non-protein binder of the present invention had a high color optical density at any concentration. As a result of microscopic observation, it was found that in the sample of the present invention, the reagent layer and the developing layer were in a separated state, whereas in the sample of the comparative example, the developing layer was sunk into the reagent layer.

[Table] Example 2 Using the copolymer of the present invention and the acrylamide-N-vinyl-2-pyrrolidone copolymer described in JP-A-58-77664 (same as Example 1), Apply Biuret reagent such as
Laminated with plain weave fabric made of PET spun yarn (equivalent to 100S). When the prepared total protein analysis element was immersed in hot water at 40°C, it was found that the reagent layer of the sample of the present invention did not dissolve, but the reagent layer of the sample of the comparative example began to dissolve.

[Table] Next, 10μ of physiological saline containing bovine albumin (BSA) with different concentrations was placed on the prepared sample.
Color density was measured at 540 nm. Similar to Example 1,
The concentration of the sample of the present invention was higher than that of the comparative sample, but it was found that this was due to the fact that the reagent layer was hardened, so the film quality was strong, and the spreading layer did not sink into the reagent layer. Next, a synthesis example of a methallyl alcohol copolymer will be shown. Synthesis Example 1 In a reaction vessel, 220ml of methanol, 80ml of distilled water,
30 g of acrylamide, 30 g of N-vinyl-2-pyrrolidone and 5 g of methallyl alcohol were added, and the mixture was sufficiently replaced with nitrogen while stirring. Add 0.5 g of azobisisobutyronitrile and stir at 60°C for 2 hours. Next, the reactants are poured into acetone to precipitate the product. After thoroughly drying, the 20% (by weight) bath liquid was adjusted and the viscosity was measured with a B-type viscometer and the result was 420 cp.
(40℃). In addition, the elemental analysis value is C: 57.85
%, H: 7.70%, N: 15.64%. Formalin, sodium dodecylbenzenesulfonate (SDS), and hydrochloric acid as a catalyst were added to this copolymer solution, and the mixture was coated on a 170μ polyethylene terephthalate support to a thickness of 26μ, and heated at 60°C.
Dry for 20 minutes. The equilibrium swelling film thickness of each sample was measured at 37°C using a swelling meter, and the swelling ratio relative to the original film thickness was determined. For samples with formalin added as a hardening agent, the swelling rate was lower as the amount of formalin added was lower, indicating that hardening was progressing, whereas samples without formalin dissolved immediately when immersed in distilled water at 37°C. did. Therefore, it was shown that the curable resin of the present invention can be cured under mild conditions (it contains the hydroxyl group of methallyl alcohol).

〔Effect of the invention〕

In the dry analysis element of the present invention, since the non-proteinaceous hydrophilic polymer binder can be crosslinked and cured, sufficient strength can be imparted to the layer by crosslinking and curing. As a result, they are easier to manufacture, store, and use. Also,
When used for total protein or albumin analysis, it was found that the color optical density was high and both measurement accuracy and sensitivity were improved.

Claims (1)

[Claims] 1. Dry integrated multilayer analysis containing a copolymer of methallyl alcohol and acrylamide or its derivatives, acrylic acid or its derivatives, or N-vinyl-2-pyrrolidone as a hydrophilic polymer binder in at least one layer. element. 2. The multilayer analytical element according to claim 1, wherein the hydrophilic polymer binder is the crosslinked copolymer. 3. The multilayer analytical element according to claim 1, wherein the hydrophilic polymer binder is the copolymer that is not crosslinked. 4. The multilayer analytical element according to claim 1, wherein the copolymer is obtained by copolymerizing methallyl alcohol and acrylamide or N-vinyl-2-pyrrolidone. 5. The multilayer analytical element according to claim 1, wherein the analyte is albumin. 6 Claim 1 in which the analyte is total protein
Multi-layer analysis elements described in .