JPH10206419A - Blood chemical analysis material - Google Patents

Abstract

PROBLEM TO BE SOLVED: To determine a plurality of specific constituents in plasma easily and accurately at a time by arranging a plurality of reagent parts that are independent of a blood-separating part flatly on a support body. SOLUTION: A blood-separating part 2 is provided on a support 1, and a plurality of reagent independent parts 3 are arranged at nearly equal distances from the center 4 where blood is dripped. Also, a recessed part or a small hole reaching the support body 1 can be provided to receive blood at the blood- separating part 2. The support 1 transmits light but does not permeate water and is made of, for example, polystyrene, polyethylenetelephthalate, and polybutylenetelephtharate. The reagent part 3 reacts with a specific constituent in blood that is filtered by the blood-separating part 2 and preferably consists of a reagent that reacts with a specific constituent in plasma that is filtered by the blood-separating part 2, has an absorbance for a wavelength of 200-900nm, and develops color or a reagent for emitting light with a wavelength of 200-900nm, and a hydrophilic carrier that retains the reagent.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a blood chemistry analysis material capable of simultaneously quantifying a plurality of specific components in blood.

[0002]

2. Description of the Related Art In recent years, in the field of medicine, the viewpoint has changed from treating diseases to preventing diseases. In addition, the growing old-age population and falling birth rates have raised people's interest in maintaining and improving their health. Judgment based on various clinical test results is indispensable for grasping a health condition, early detection of a disease, and prediction of a future disease. Above all, the blood test is sugar in the blood,
It is particularly important because it quantifies chemical substances such as lipids, proteins, inorganic ions, enzymes and hormones. Heretofore, blood tests have been performed by a method of reacting with a reagent solution prepared in advance using plasma or serum (called wet chemistry). This method based on a reaction in a solution is convenient when processing a large number of specimens at once, but it is useful when a laboratory technician is required to obtain results on the spot, such as an emergency test, or at night or on a holiday. Is not suitable for responding when the user is absent. In addition, even in a facility with a small sample amount, the inspection by wet chemistry is often inconvenient. In addition, the need for `` tests closer to the patient, '' such as outpatient blood tests prior to medical examinations by doctors, bedside tests for daily management of inpatients, and home tests, is pointed out as the direction of medical treatment. Therefore, wet chemistry is not suitable for inspection for this purpose.

In contrast to wet chemistry, an analysis method called dry chemistry using a dry blood chemistry analysis material has been developed against the background of the above-mentioned trends in medical sites, and has been rapidly applied to clinical sites in Japan, the United States and Europe. It is a new blood test system that is becoming more and more popular. Today, dry chemistry is mainly analyzed using a multilayer film having a structure as shown in FIG. The blood spotted on the upper part of the developing layer a is horizontally spread in the developing layer a, and at the same time, the blood cells are filtered, and only the plasma reaches the reagent layer c through the reflective layer b. In the reagent layer c, a reaction occurs between the reagent and the test substance in the plasma, and a dye is generated in proportion to the concentration of the test substance. The amount of the dye is measured by a spectrophotometer or the like. In dry chemistry, the amount of the dye is reflected and measured through the lowermost transparent support d. Therefore, the reflection layer b in FIG. 1 functions as a reflection plate that blocks the color of red blood cells.

[0004] Components in blood measured by dry chemistry include inorganic ions such as glucose, urea nitrogen, uric acid, cholesterol, triglyceride, calcium, and phosphorus, albumin, glutamate oxaloacetate transaminase (GOT), and glutamate pyruvate transaminase. (GPT), lactate dehydrogenase (LD
H), etc., but the analysis using whole blood containing blood cell components is limited to several components because it is difficult to obtain a reflective layer material with excellent light shielding properties. Most are methods using plasma or serum from which blood cell components have been removed.

[0005] In the case of dry chemistry using a multilayer film type analysis material, only one component can be analyzed by one film due to the structure of the multilayer film. JP-A-6-242107 discloses a dry analytical element for multi-item measurement in which a single film can analyze a plurality of components. This analysis element is composed of a plurality of partial analysis elements in which a hydrophilic polymer layer and a porous spreading layer are laminated on a water-impermeable support, and a blood cell separation element laid on the water-impervious support in a detachable manner. Consists of At the time of measurement, the blood spotted on the blood cell separation element is filtered by blood cells, and the plasma is spread to each partial analysis element. Thereafter, the blood cell separation element is peeled and removed, and a measurement reagent is spotted on each partial analysis element for analysis. This method requires an operation such as detachment of a blood cell separation element, and in Japanese Patent Application Laid-Open No. 6-242107, the purpose is to perform spotting and analysis of a measurement reagent at a laboratory, and therefore dry chemistry is originally used. This does not meet the target of “examination closer to the patient”.

[0006]

SUMMARY OF THE INVENTION An object of the present invention is to provide a blood chemistry analysis material which can easily and accurately quantify a plurality of specific components in blood at one time.

[0007] The present inventors have proposed that, on a support, an appropriate number of reagent sections independent of the blood cell separation section, or an appropriate number of independent blood cell separation sections and reagent sections corresponding to the number of the blood cell separation sections are placed on a flat surface. The present inventors have found that the object of the present invention can be achieved by using a blood chemistry analysis material having a regularly arranged structure, and reached the present invention.

[0008]

That is, the present invention provides a blood chemistry analysis material characterized in that a blood cell separation section and an appropriate number of independent reagent sections are arranged in a plane on a support. And

Further, in the blood cell chemical analysis material of the present invention, an appropriate number of the independent blood cell separation sections and a number of the reagent sections corresponding to the number of the blood cell separation sections are planarly arranged on the support. It is characterized by becoming.

Further, the blood cell chemical analysis material of the present invention is characterized in that the plurality of reagent parts are arranged at substantially the same position from the blood dropping part.

Further, the blood cell analysis material of the present invention is characterized in that the blood cell separation section is made of a porous substance.

Further, in the blood cell chemical analysis material of the present invention, the reagent which reacts with a component in blood to exhibit a color having an absorbance at 200 to 900 nm or 200 to 900
It is characterized by comprising a reagent that emits light of nm and a transparent hydrophilic carrier holding the reagent.

[0013]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS A structural example of a blood chemistry analysis material of the present invention will be described with reference to FIGS. 2 and 3 and FIGS. 4 and 5. FIG. FIG. 2 is a plan view showing a specific example of the blood cell chemical analysis material of the present invention, and FIG. 3 is a sectional view taken along the line XY of FIG.
FIG. 4 is a plan view showing another specific example of the blood cell analysis material of the present invention, and FIG. 5 is a sectional view taken along the line XY of FIG. As shown in FIGS. 2 and 3, the blood chemistry analysis material of the present invention is preferably formed by stacking a blood cell separation part 2 on a support 1 and further from the center 4 of the blood chemistry analysis material onto which blood is dropped. It has a structure in which an appropriate number of independent reagent units 3 are arranged at substantially the same position and two-dimensionally. The blood cell separation unit 2
Is provided with a recess or support 1 on its upper surface to receive blood.
Can be provided. Further, as shown in FIG. 4 and FIG. 5, the blood chemistry analysis material of the present invention has an appropriate number of independent blood cell separation sections 2 arranged on a support 1 on a radiator. This is a structure in which the reagent section 3 is arranged in a plane. In the above, an appropriate number of independent reagent parts 3
And the number of independent blood cell separation units 2 are determined arbitrarily according to the number of specific components in the blood to be analyzed. With such a configuration, it is possible to analyze a desired number of specific components in blood with one blood chemistry analysis material.

The above support is light-permeable and water-impermeable, for example, polystyrene, polyethylene terephthalate, polybutylene terephthalate, cellulose ester (eg, cellulose diacetate, cellulose triacetate, cellulose acetate propionate), bisphenol A A known water-impermeable transparent support having a thickness of 50 μm to 2 mm, such as a film of polycarbonate, polymethyl methacrylate, polylactic acid or the like, a glass plate, a quartz plate, or the like can be used.

The blood cell separation section serves to filter blood cells from the dropped blood and supply only plasma to the reagent section. In the present invention, as shown in FIGS. 2 and 3, one blood cell separation unit 2 can be arranged so as to surround the entire reagent unit 3, and as shown in FIGS. 4 and 5, It is also possible to arrange independent blood cell separation units 2 corresponding to the number of independent reagent units 3. In the former case,
Blood is dropped directly on the surface of the blood cell separation section 2, but in the latter, the center of the blood chemistry analysis material onto which blood is dropped is:
It becomes a space surrounded by the blood cell separation unit 2 and serves as a pool for filtration by the blood cell separation unit 2. In this case, in order to prevent hemolysis when blood comes into contact, the surface of the support 1 at this portion may be subjected to hemolysis prevention processing by a known method using a hemolysis inhibitor such as di-2-ethylhexyl phthalate. it can. In order to reduce the amount of blood remaining in this portion, the surface of the support may be subjected to a water-repellent treatment by a known method using an organopolysiloxane or a fluoroalkyl compound.

In the present invention, a fibrous or non-fibrous porous material having a hydrophilic surface is used for the blood cell separation part. As a fibrous porous material, for example,
Examples include woven fabric, glass fiber, cellulose fiber, paper and the like which have been subjected to a hydrophilic treatment described in JP-B-61-61347. In addition, as a non-fibrous porous substance,
JP-B-53-21677, U.S. Pat. No. 1,42
No. 1,341, for example, cellulose esters such as cellulose acetate, cellulose acetate / butyrate, a brush polymer composed of cellulose acetate (general name: membrane filter), polyamides such as 6-nylon and 6,6-nylon ,polyethylene,
Microporous materials such as polypropylene, small polymer particles, glass particles, diatomaceous earth, and the like can be mentioned as porous materials having continuous voids in which hydrophilic or non-water-absorbing polymers are combined.

The degree of porosity of the above-mentioned porous substance cannot be unconditionally determined because it varies depending on the degree of hydrophilicity of the surface, the state of porosity, the shape of pores, the manner of distribution of pores, etc. In the case of a porous material, the average pore size is 0.1
In the case of a fibrous porous substance, it is represented by a cotton count, and preferably has a count of 10 to 100, and preferably has a count of 20 to 80. The range corresponds to the voids of a plain fabric such as a cotton yarn broad.

The porous substance is preferably in the form of a film, and a film from which a reagent portion has been previously removed or a film cut so as to form an independent blood cell separation portion is supported before the reagent portion is prepared. Laminating on the body,
This is important for easily obtaining the blood chemistry analysis material of the present invention. Further, a film made of a porous substance may be simply laminated on the support, but more preferably both are adhered with an adhesive. As the adhesive to be used, various adhesives described in JP-A-62-138756, and others, for example, "Printing Engineering Handbook" edited by The Printing Society of Japan (Gihodo Publishing Co., Ltd., 1983) 839-
A known adhesive described on page 853 can be used. Furthermore, it is also possible to stick both using a double-sided tape.

When the porous substance is not in the form of a film, a coating agent for forming a porous blood cell separating portion, for example, acetone-dichloroethane (1: 1) of cellulose acetate.
1) mixed solvent solution, silica gel, microcrystalline cellulose,
A coating agent prepared by dispersing 80 to 120 mesh glass beads in a small amount of starch paste aqueous solution or gelatin solution is coated on a support and dried, leaving a part to be a reagent part. It is also possible to adopt a method of stacking. Also in this case, it is desirable to form the blood cell separation section before preparing the reagent section.

It is desirable that the thickness of the blood cell separation part is 5 to 500 μm, preferably 10 to 250 μm. If the thickness of the blood cell separation section is less than 5 μm, it is difficult to sufficiently perform filtration and separation of blood cells, which is not preferable. Also,
If the thickness of the blood cell separation portion is greater than 500 μm, plasma is retained in the blood cell separation portion, and a large amount of blood is required, which is not preferable.

The blood cell separation section may contain a nonionic, anionic, cationic or amphoteric surfactant in order to promote the spread of plasma. Further, for the purpose of controlling the development property, a development control agent such as a hydrophilic polymer may be included. Furthermore, to promote the detection reaction in the reagent section, or reduce interference and interference reactions,
Various reagents or some of the reagents for blocking can be included.

The reagent section is a reagent which reacts with a specific component in the plasma filtered by the blood cell separation section to exhibit a color having a absorbance at a wavelength in the range of 200 to 900 nm or 20.
Those comprising a reagent emitting light of 0 to 900 nm and a transparent hydrophilic carrier holding the reagent are preferred.

As the specific component, for example, glucose,
Chemicals such as urea nitrogen, creatinine, uric acid, total cholesterol, high density lipoprotein (HDL) cholesterol, triglyceride, total bilirubin, calcium, inorganic phosphorus, total protein, albumin, ammonia, hemoglobin, and γ-glutamyl transpeptidase (γ -GTP), glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT), creatine phosphokinase, lactate dehydrogenase (LDH), alkaline phosphatase (ALP), amylase, leucine aminopeptidase, and other enzymes. be able to. In the reagent section, these specific components react with the reagents, and depending on the amount of the components in the plasma, exhibit a color having an absorbance of 200 to 900 nm or emit light of 200 to 900 nm, so that these should be measured. Thus, the components in the plasma can be quantified.

As a reagent for quantifying the above-mentioned specific component, a reagent known in the aforementioned wet chemistry can be used. This reagent may contain enzymes such as glucose oxidase, uricase, cholesterol esterase, cholesterol oxidase, glucose-6-phosphate dehydrogenase, peroxidase, and diaphorase. Also, in order to improve the measurement sensitivity,
It is also possible to increase the concentration of the reagent. Furthermore, in wet chemistry, a reaction that produces color in the visible region due to the formation of quinone dyes, chelates, diformazan, and the like is mainly used, but luminol, lucigenin, bis (2,4,6-trichlorophenyl) oxalate (T
It is also possible to use a method of generating chemiluminescence using (CPO) or the like.

As the transparent hydrophilic carrier for holding the above reagent, known carriers, for example, natural polymer compounds such as gelatin, albumin, collagen, agar, agarose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, polyacrylamide And a synthetic polymer compound such as polyacrylic acid.

In the present invention, a radiation-curable curable liquid resin composition described below can be used as a carrier for holding the above reagent. Since the curable liquid resin composition can be cured at a high speed by irradiating radiation, the drying step can be omitted as compared with a conventionally used hydrophilic carrier. Also,
The reagent part obtained by using the curable liquid resin composition is excellent in water absorption and reactivity between the reagent and the specific component, so that measurement sensitivity can be improved.

The above-mentioned curable liquid resin composition is composed of 100 parts by weight of a (meth) acrylate liquid resin (A) and a number average molecular weight of 1,000 having an unsaturated double bond in the molecule.
The following (meth) acrylate monomers (B) 1-1,0
The (meth) acrylate-based liquid resin (A) is an alkylene glycol (meth) acrylate-based monomer (a-1) 20 represented by the following general formula (1).
And a polymerizable monomer (a-2) excluding the monomer (a-1) of from 0 to 80% by weight and having a number average molecular weight of 10,000 to 200. , 0
It is a solventless liquid resin composition having a viscosity of 1 to 10,000 poise (50 ° C.).

CH ₂ ＣC (R ¹ ) COO (C _n H _2n O) _m R ² (1) (wherein, R ¹ is a hydrogen atom or a methyl group, and R ² is a carbon atom having 1 carbon atom.
An alkyl group or a phenyl group of 5 to 5, n is an integer of 1 to 3,
m represents an integer of 3 to 25, respectively. )

(Meth) acrylate liquid resin (A)
Is a solvent-free liquid resin obtained by copolymerizing an alkylene glycol (meth) acrylate monomer (a-1) and the polymerizable monomer (a-2), and the resin composition is liquefied. This gives a suitable viscosity at the time of preparing the reagent part, and when the reagent contains an enzyme or the like, it is also a component for causing these to stably exist in the reagent part. Further, it plays a role of imparting toughness and flexibility to the reagent part after curing.

In the present invention, the alkylene glycol (meth) acrylate monomer (a-1) represented by the general formula (1) is used because the (meth) acrylate resin (A) is in a liquid state. . As the monomer (a-1), for example, methoxytetraethylene glycol (meth) acrylate, ethoxytetraethylene glycol (meth) acrylate, propoxytetraethylene glycol (meth) acrylate, n-butoxytetraethylene glycol (meth) acrylate, n-pentyloxytetraethylene glycol (meth) acrylate,
Tetrapropylene glycol (meth) acrylate, methoxytetrapropylene glycol (meth) acrylate, ethoxytetrapropylene glycol (meth) acrylate, propoxytetrapropylene glycol (meth) acrylate, n-butoxytetrapropylene glycol (meth) acrylate, n-pentyloxy Tetrapropylene glycol (meth) acrylate, methoxy polyethylene glycol (meth) acrylate, ethoxy polyethylene glycol (meth) acrylate, or phenoxytetraethylene glycol (meth) acrylate, phenoxyhexaethylene glycol (meth)
Examples thereof include acrylate, phenoxy polyethylene glycol (meth) acrylate, and phenoxytetrapropylene glycol (meth) acrylate.
Among them, m in the general formula (1) is particularly 3 to 2
By using the monomer (a-1) having a polyoxyalkylene side chain of 5, preferably 4 to 22, the viscosity of the copolymer can be effectively reduced. Further, enzymes and the like in the reagent are stably retained by the obtained copolymer. Furthermore, when curing by irradiation of radiation,
The crosslinking reaction between the polyoxyalkylene side chains proceeds effectively. When m is less than 3, it is difficult to obtain a low-viscosity liquid resin, and when m exceeds 25, it is difficult to increase the degree of polymerization, and the obtained liquid resin becomes solid, making it difficult to prepare the reagent portion, which is not preferable.

Such a monomer (a-1) contains 2
It is desirably contained in an amount of 0 to 100% by weight, preferably 40 to 95% by weight. When the monomer (a-1) in the copolymer is 4
When the amount is less than 0% by weight, particularly less than 20% by weight, it becomes difficult to maintain a preferable viscosity. In addition, a problem arises in stability in a copolymer such as an enzyme.

In the present invention, the polymerizable monomer (a-2) excluding the monomer (a-1) is used for improving the physical properties of the cured reagent part. The polymerizable monomer includes a radical polymerizable monomer having a carboxyl group, an amide group or a hydroxyl group in a molecule, and other polymerizable vinyl monomers.

The radically polymerizable monomer having a carboxyl group in the molecule includes, for example, maleic anhydride, maleic acid, fumaric acid, itaconic acid, citraconic acid, or their alkyl or alkenyl monoester, phthalic acid β -(Meth) acryloxyethyl monoester, isophthalic acid β- (meth) acryloxyethyl monoester, terephthalic acid β- (meth) acryloxyethyl monoester, succinic acid β- (meth) acryloxyethyl monoester, acrylic Acids, methacrylic acid, crotonic acid, cinnamic acid and the like can be mentioned.

Examples of the radical polymerizable monomer having an amide group in the molecule include (meth) acrylamide, N
-Methylol (meth) acrylamide, N-methoxymethyl (meth) acrylamide, N-ethoxymethyl (meth) acrylamide, N-propoxymethyl (meth) acrylamide, N-butoxymethyl (meth) acrylamide, N-pentyloxymethyl (meth) ) Monoalkylol (meth) acrylamide such as acrylamide,
N, N-di (methylol) (meth) acrylamide, N
-Methylol-N-methoxymethyl (meth) acrylamide, N, N-di (methoxymethyl) (meth) acrylamide, N-ethoxymethyl-N-methoxymethyl (meth) acrylamide, N, N-di (ethoxymethyl)
(Meth) acrylamide, N-ethoxymethyl-N-propoxymethyl (meth) acrylamide, N, N-di (propoxymethyl) (meth) acrylamide, N-butoxymethyl-N- (propoxymethyl) (meth) acrylamide, N N, N-di (butoxymethyl) (meth) acrylamide, N-butoxymethyl-N- (methoxymethyl) (meth) acrylamide, N, N-di (pentyloxymethyl) (meth) acrylamide, N-methoxymethyl-N Dialkylol (meth) acrylamide such as-(pentyloxymethyl) (meth) acrylamide.

The radical polymerizable monomer having a hydroxyl group in the molecule includes, for example, 2-hydroxyethyl (meth)
Acrylate, 3-hydroxypropyl (meth) acrylate, 4-hydroxybutyl (meth) acrylate,
(Meth) acrylates having a hydroxyl group such as ethylene glycol (meth) acrylate, propylene glycol (meth) acrylate, tetraethylene glycol (meth) acrylate, tetrapropylene glycol (meth) acrylate, and polyethylene glycol (meth) acrylate; N- (hydroxy Examples thereof include (meth) acrylamide having a hydroxyl group such as methyl) (meth) acrylamide, N- (hydroxyethyl) (meth) acrylamide, and diacetone (meth) acrylamide. Also, for example, allyl alcohol, 4-hydroxy-1-butene, 4-hydroxymethylstyrene, 4-hydroxyethylstyrene, 1,4-dihydroxy-2-
A vinyl monomer having a primary hydroxyl group such as butene can also be used.

Other polymerizable vinyl monomers include, for example, aromatic monomers such as styrene and vinyltoluene;
Methyl (meth) acrylate, ethyl (meth) acrylate, butyl (meth) acrylate, (meth) acrylate having an alkyl group such as 2-ethylhexyl (meth) acrylate, methoxydiethylene glycol (meth) acrylate, phenoxydiethylene glycol (meth) acrylate, Examples include alkoxy groups such as phenoxyethylene glycol (meth) acrylate, and (meth) acrylates containing a phenoxy group.

It is desirable that the polymerizable monomer (a-2) is contained in the copolymer in an amount of 0 to 90% by weight, preferably 5 to 60% by weight. The polymerizable monomer (a-
If 2) is more than 60% by weight, especially more than 80% by weight, the viscosity of the liquid resin is undesirably high.

When the resin composition used in the present invention is cured by irradiating an electron beam, the monomer (a-
1) is preferably a compound of the formula (1) wherein R ¹ is a hydrogen atom. In this case, it is preferable that the polymerizable monomer (a-2) used in the copolymer does not have a quaternary carbon in the main chain when copolymerized, such as an acrylate monomer or styrene.

The (meth) acrylate liquid resin (A) used in the present invention has a number average molecular weight of 10,000 to 20.
000, preferably 11,000 to 100,000
It is desirable that If the number average molecular weight is smaller than the above-mentioned value, it is difficult to isolate the liquid resin from the polymerization solvent, and at the same time, the physical properties of the reagent part after curing are undesirably reduced. On the other hand, when the number average molecular weight is larger than the above value, it is not preferable because a large amount of a low molecular weight compound needs to be added to lower the viscosity of the liquid resin.

(Meth) acrylate liquid resin (A)
Is a method of dissolving a mixture of the above monomers (a-1) and (a-2) in a solvent in the presence of a radical polymerization initiator, or a method of dissolving the above monomers (a-1) and (a- It can be produced by radical polymerization using the method of dropping the mixture of 2). Examples of the radical polymerization initiator include benzoyl peroxide, t-butyl peroxide, cumene hydroperoxide, lauroyl peroxide, and other organic peroxides (for example, Taiseisha “Crosslinking Agent Handbook”, p.
Known compounds such as peroxides described in No. 35), azo compounds such as azobisisobutyronitrile and azobiscyclohexanenitrile, and persulfate initiators such as potassium persulfate and ammonium persulfate can be used.

Examples of the solvent used for the polymerization include ethyl acetate, toluene, methyl ethyl ketone, benzene, dioxane, n-propanol, methanol, isopropanol, tetrahydrofuran, n-butanol, sec-
-Butanol, tert-butanol, isobutanol, methyl cellosolve, butyl cellosolve, methyl carbitol, ethyl carbitol, methyl cellosolve acetate, ethyl cellosolve acetate, diacetone alcohol and the like.

After the copolymerization reaction, the solvent used was purified by precipitation,
The solvent-free (meth) acrylate-based liquid resin (A) is obtained by removing by a method such as distillation. The obtained liquid resin is
It is desirable that the viscosity at 50 ° C. is 1 to 10,000 poise, preferably 10 to 1,000 poise. If the viscosity is lower than 1 poise, the reagent portion after curing becomes brittle, which is not preferable. Conversely, when the viscosity is higher than 10,000 poise, a large amount of radiation is required for curing, and a large amount of (meth) acrylate-based monomer (B) is required for lowering the viscosity.
Is not preferred because

In the present invention, the (meth) acrylate monomer (B) having an unsaturated double bond in the molecule and having a number average molecular weight of 1,000 or less is mixed with the (meth) acrylate liquid resin (A). Then, it is used to adjust its viscosity and curability.

Examples of the (meth) acrylate monomer (B) include, for example, methyl (meth) acrylate, butyl (meth) acrylate, 2-hydroxyethyl (meth)
Monofunctional (meth) acrylate monomers such as acrylate, phenoxymethyl (meth) acrylate, phenoxyethyl (meth) acrylate, and benzyl (meth) acrylate; ethylene glycol di (meth) acrylate; diethylene glycol di (meth) acrylate; Ethylene glycol di (meth) acrylate, polyethylene glycol di (meth) acrylate, 1,3-butanediol di (meth) acrylate, 1,6-hexanediol di (meth) acrylate, 1,9-nonanediol di (meth) Acrylate, neopentyl glycol di (meth) acrylate, 2,2-bis [4-{(meth) acryloxy ethoxy} phenyl] propane,
2,2-bis [4-{(meth) acryloxydiethoxy {phenyl] propane, 2,2-bis [4-{(meth) acryloxypolyethoxy {phenyl] propane, 2,2-bis [4-} ( (Meth) acryloxypropoxydiphenyl] propane, 2,2-bis [4-
{(Meth) acryloxy dipropoxy} phenyl] propane, 2,2-bis [4-} (meth) acryloxy.
Bifunctional (meth) acrylate monomers such as polypropoxydiphenyl] propane, trimethylolpropanetri (meth) acrylate, tetramethylolmethanetri (meth) acrylate, tetramethylolethanetri (meth) acrylate, tetramethylolmethanetetra Examples thereof include tri- or higher-functional (meth) acrylate monomers such as (meth) acrylate, ethylene oxide-modified trimethylolpropane tri (meth) acrylate, and dipentaerythritol hexa (meth) acrylate. Among them, the use of a (meth) acrylate monomer having a polyoxyalkylene moiety is preferable because the crosslinking reaction of the polyoxyalkylene moiety effectively proceeds when the composition is cured by irradiation with radiation. In addition, the polyoxyalkylene moiety contributes to stabilization of an enzyme or the like in the reagent portion.

The (meth) acrylate monomer (B)
It is desirable that the viscosity at 50 ° C. is 0.01 to 60 poise, preferably 0.1 to 50 poise. If the viscosity is lower than this range, many of low molecular weight, reagents, especially if the enzyme is included in the reagent, the effect on the enzyme in the reagent is large, conversely, if the viscosity is higher than this range,
It is not preferable because the role of the (meth) acrylate liquid resin as a viscosity modifier becomes poor.

In the present invention, the (meth) acrylate liquid resin (A) and the (meth) acrylate monomer (B)
Is mixed with 100 parts by weight of the (meth) acrylate-based liquid resin (A), from 1 to 1,000 parts by weight, preferably from 2 to 50 parts by weight of the (meth) acrylate-based monomer (B).
It is desirably 0 parts by weight. If the compounding ratio is less than this, the change in viscosity of the resin composition is poor, and if the compounding ratio is more than this, the amount of residual monomer after curing increases, the volume shrinkage during curing is remarkable, the cured reagent part becomes brittle It is not preferable for such reasons.

The reagent part in the blood chemistry analysis material of the present invention is obtained by mixing the above-mentioned reagent with a known hydrophilic carrier or the above-mentioned curable liquid resin composition and supporting the mixture other than the blood cell separation part formed in advance. It can be obtained by drying or curing after application to a predetermined place on the body. The reagent is generally prepared as a solution, and the amount added in this case is desirably 0.1 to 1,000 parts by weight based on 100 parts by weight of the hydrophilic carrier or the curable liquid resin composition. Also, in order to improve the physical properties of the obtained reagent part,
If necessary, a compatibilizer and a surfactant may be added.
The amount of these additives is preferably 20 parts by weight or less, more preferably 10 parts by weight or less, based on 100 parts by weight of the hydrophilic carrier or the curable liquid resin composition.

A known hydrophilic carrier not containing the above-mentioned reagent or the above-mentioned curable liquid resin composition is applied to a place on the support where a reagent portion is to be formed, and then dried or cured. It is also possible to obtain a reagent part by impregnation. In this case, the addition amount of the reagent solution is preferably 0.1 to 1,000 parts by weight based on 100 parts by weight of the hydrophilic carrier or the curable liquid resin composition.

The curing of the above curable liquid resin composition is carried out by using X
This is achieved by irradiating the composition with radiation such as radiation, gamma rays, electron beams, ultraviolet rays, visible rays, infrared rays, and the like. When a reagent part is obtained by curing by irradiation with an electron beam, preferably 10 to 1,000 KeV, more preferably 30 to 30 KeV.
It is desirable to use an electron beam irradiation device having an energy in the range of 0 KeV. Electron beam irradiation dose (DOSE)
Is preferably in the range of 0.1 to 100 Mrad, more preferably 0.5 to 20 Mrad.
If the irradiation dose is smaller than this, it is difficult to obtain a sufficiently cured product, and if it is larger than this, the damage to the reagent and the liquid resin is undesirably increased.

It is desirable that the thickness of the reagent part is 5 to 500 μm, preferably 10 to 250 μm. If the thickness of the reagent portion is smaller than this, the amount of color development or the amount of light emission in the reagent portion is decreased, which leads to a decrease in measurement sensitivity, which is not preferable. On the other hand, if the thickness of the reagent portion is larger than this, a large amount of blood is required to generate sufficient and uniform color or light emission, which is not preferable.

In the blood chemistry analysis material of the present invention, a water-impermeable film or the like may be laminated thereon for the purpose of protecting the blood cell separation section and the reagent section. As a film to be laminated, a film having the same composition as the support, that is, polystyrene, polyethylene terephthalate, polybutylene terephthalate, cellulose ester (cellulose diacetate, cellulose triacetate, cellulose acetate propionate, etc.), polycarbonate of bisphenol A, polymethyl methacrylate And polylactic acid. Further, a glass plate, a quartz plate, or the like can be used.

As a method for measuring dry chemistry, a reflection optical system is usually used. However, in the method using the blood chemistry analysis material of the present invention, measurement is performed by a transmission photometric method through a support through a support. Is possible, which allows for a reduction in dry chemistry using conventional materials.
Measurement sensitivity can be improved. Further, as compared with a measuring device using a reflection optical system, an integrating sphere or the like is not required, so that the measuring device can be simplified. Further, by laminating a light-impermeable film or the like on the reagent part, it is possible to measure the reflected light from the support side, similarly to the conventional dry chemistry.

In the blood chemistry analysis material of the present invention, there is no problem if the number of reagent parts is one or more. However, for the convenience of preparing blood chemistry analysis materials,
Preferably, the number of reagent parts is in the range of 2-8.

The blood chemistry analysis material of the present invention is intended to analyze a specific component in blood collected by a subject himself mainly using a puncture needle or the like. Therefore, the amount of blood dropped on the blood chemistry analysis material is as small as about 5 to 200 μl. Therefore, it is desirable that the blood chemistry analysis material be small in order to spread the plasma throughout the reagent section. Specifically, in FIGS. 2 and 4, the length of one side is desirably 0.5 to 2.0 cm.

[0054]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail with reference to embodiments. The embodiment is an example of a blood chemistry analysis material having six reagent parts, but the present invention is not limited to these. A method for measuring the number average molecular weight and the viscosity in this example is shown below. 1) Number average molecular weight: The value in terms of polystyrene measured by gel permeation chromatography (manufactured by Tosoh Corporation: SC-8020) was used. 2) Viscosity: Rheometer (Rheometrics: RDS)
-Type II and RFS-II type rheometer), and the value of the steady flow viscosity at a shear rate of 1 to 10 sec-1 was used. ◎ The electron beam irradiation apparatus and irradiation conditions are shown below. 1) Area beam type electron beam irradiator (manufactured by Nissin High Voltage: Curetron EBC-200-20-3)
0) Electron beam acceleration: 200 KeV DOSE: Adjusted by current amount 2) MIN-EB (manufactured by AIT) Electron beam acceleration: 60 KeV DOSE: Adjusted by belt conveyor speed

(Synthesis Example 1) (Meth) acrylate liquid resin (1) In a 500 ml four-necked round-bottom flask equipped with a stirrer, a nitrogen inlet tube, a temperature sensor and a condenser, methoxypolyethylene glycol acrylate (as described above) was used as a monomer. In the general formula (1), m = 9, and isopropanol as a solvent (the monomer concentration at the time of preparation is
% By weight) and azobisisobutyronitrile (AIB)
N) as initiator (1% by weight of the total monomer concentration)
After reacting in a hot water bath at 6 ° C. for 6 hours, 0.1 wt% of AIBN was further added, and heating and stirring were continued for 2 hours. After the reaction, a diversion tube was attached between the reactor and the condenser, the temperature of the hot water bath was raised to 95 ° C., the solvent was distilled off while stirring at normal pressure, and the pressure was reduced to 40 mmHg or less under the same temperature conditions. The solvent was completely distilled off to obtain a viscous liquid resin (1). The number average molecular weight of the obtained liquid resin is 22,100,
The viscosity at 50 ° C. was 132 poise.

(Synthesis Example 2) (Meth) acrylate liquid resin (2) Except that methoxytetraethylene glycol acrylate, 4-hydroxybutyl acrylate and styrene (weight ratio = 75: 20: 5) were used as monomers. The same operation as in Synthesis Example 1 was performed to obtain a viscous liquid resin (2). The number average molecular weight of the obtained liquid resin was 16,100,
The viscosity at 50 ° C. was 163 poise.

(Synthesis Example 3) (Meth) acrylate liquid resin (3) As monomers, phenoxypolyethylene glycol acrylate (m = 9 in the above formula (1)) and acrylic acid (weight ratio = 95: 5) ) Was carried out in the same manner as in Synthesis Example 1 except that) was used to obtain a viscous liquid resin (3). The number average molecular weight of the obtained liquid resin is 2
The viscosity at 0,400 and 50 ° C. was 7,430 poise.

(Synthesis Example 4) (Meth) acrylate liquid resin (4) Synthesis Example 1 was repeated except that methoxypolyethylene glycol methacrylate (where m = 9 in the above formula (1)) was used as a monomer. The same operation was performed to obtain a viscous liquid resin (4). The number average molecular weight of the obtained liquid resin was 23,000, and the viscosity at 50 ° C. was 145 poise.

(Example 1) <1. Preparation of blood chemistry analysis material> 1-1. Preparation of Support Having Laminated Blood Cell Separation Section A cellulose fiber filter paper having an average pore diameter of 2.5 μm and a thickness of 100 μm was cut out of the reagent section so as to have the shape shown in FIGS. 2 and 3. A 200 μm polyethylene terephthalate (PET) colorless transparent smooth sheet was adhered and laminated using a heat-adhesive hot-melt adhesive. The length of one side of the obtained PET colorless transparent smooth sheet was 1.0 cm. 1-2. Preparation of Reagent for Glucose Measurement Each component was dissolved in 60 mM phosphate buffer (pH 7.1) to prepare a reagent for glucose measurement having the composition shown in the following table.

TABLE 1 Glucose oxidase 90 units / ml Peroxidase 6.5 units / ml Phenol 53 mmol / l 4-aminoantipyrine 5.0 mmol / l 1-3. Preparation of reagent for measuring uric acid Each component was dissolved in a 50 mM phosphate buffer (pH 6.0) to prepare a reagent for measuring uric acid having the composition shown in the following table.

Uricase 0.9 units / ml peroxidase 63 units / ml 4-aminoantipyrine 7.2 mmol / l 3-methyl-N-ethyl-N- (β-hydroxyethyl) -aniline 56 mmol / l 1-4. Preparation of Reagent for Measurement of Total Cholesterol Each component was dissolved in 150 mM Tris buffer (pH 7.0) to prepare a reagent for measurement of total cholesterol having the composition shown in the following table.

Table 3 Cholesterol esterase 5.6 units / ml Cholesterol oxidase 1.2 units / ml Peroxidase 21 units / ml 4-aminoantipyrine 7.3 mmol / l p-chlorophenol 76 mmol / l 1-5. Preparation of reagent for calcium measurement 880 mM monoethanolamine buffer (pH 11.
Each component was dissolved in 0) to prepare a calcium measurement reagent having the composition shown in the following table.

Table 4 o-Cresolphthalein complexone 0.7 mmol / l 8-quinolinol 69 mmol / l 1-6. Preparation of Reagent for Measurement of Albumin Each component was dissolved in a 75 mM succinate buffer (pH 4.2) to prepare a reagent for measurement of albumin having the composition shown in the following table.

Table 5 Bromcresol Green 1.7 mmol / l 1-7. Preparation of Reagent for Measurement of Alkaline Phosphatase Each component was dissolved in 50 mM carbonate buffer (pH 10.2) to prepare a reagent for measurement of alkaline phosphatase having the composition shown in the following table.

Table 6: Phenyl phosphate 37 mmol / l 4-aminoantipyrine 54 mmol / l potassium ferricyanide 36 mmol / l 1-8. Preparation of Reagent Part In 100 parts by weight of the above (meth) acrylate liquid resin (1), polyethylene glycol diacrylate (number average molecular weight = 50) was used as a (meth) acrylate monomer.
8) was mixed with 300 parts by weight to obtain a radiation-curable solventless liquid resin. The viscosity of this liquid resin is 25 poise (5
0 ° C). To 100 parts by weight of this liquid resin, 50 parts by weight of the above-described reagents for measuring glucose, uric acid, reagents for measuring total cholesterol, reagents for measuring calcium, reagents for measuring albumin, and reagents for measuring alkaline phosphatase were added. Mixed. The obtained mixture is applied to a portion of a PET colorless transparent smooth sheet on which a blood cell separation section is to be formed as a reagent section, and irradiated with an electron beam using an area beam type electron beam irradiation apparatus (DOSE = 5 Mrad).
And cured to obtain a blood chemistry analysis material having a shape as shown in FIG. 2 and FIG. In addition, the thickness of the reagent part after hardening was 100 micrometers, and the diameter was 0.32 cm.

<2. Measurement> Heparin-containing whole blood of a healthy individual was dropped on the blood drop portion, and incubated at 37 ° C. for 5 minutes.
The absorbance at the wavelength corresponding to each color was measured by a transmission photometric method. When the concentration of each component was calculated from a calibration curve prepared between the concentration of each component and the absorbance prepared in advance, glucose = 95 mg / dl, uric acid =
5.2 mg / dl, total cholesterol = 150 mg / d
1, calcium = 9.6 mg / dl, albumin = 4.
5 g / dl, alkaline phosphatase = 7.2 K-A
Units. As a result of repeating the same operation 10 times and examining the reproducibility, the CV (%) was determined to be glucose = 3.
1, uric acid = 2.5, total cholesterol = 6.5, calcium = 4.0, albumin = 2.0, alkaline phosphatase = 5.4, and the measurement method using the blood chemistry analysis material of the present invention is sufficient. It was confirmed that the measurement method had high reliability.

Example 2 A liquid resin (2) was used in place of the (meth) acrylate liquid resin (1).
The same operation as in Example 1 was performed to obtain a blood chemistry analysis material.
Further, using this, the same measurement as in Example 1 was performed, and the respective component concentrations and reproducibility were obtained. as a result,
Glucose = 93 mg / dl (3.0), uric acid = 5.5
mg / dl (2.5), total cholesterol = 148 mg
/ Dl (6.8), calcium = 9.6 mg / dl
(3.5), albumin = 4.4 g / dl (2.0),
Alkaline phosphatase = 6.8 KA units (5.
0) was obtained, indicating that the blood chemistry analysis material of the present invention is effective for quantifying components in blood. In addition,
The values in parentheses indicate the values of reproducibility and CV (%) (the same applies to the following examples).

Example 3 A liquid resin (3) was used in place of the (meth) acrylate liquid resin (1).
The same operation as in Example 1 was performed to obtain a blood chemistry analysis material.
Further, using this, the same measurement as in Example 1 was performed, and the respective component concentrations and reproducibility were obtained. as a result,
Glucose = 95 mg / dl (3.0), uric acid = 5.1
mg / dl (2.8), total cholesterol = 152 mg
/ Dl (6.0), calcium = 9.8 mg / dl
(3.9), albumin = 4.5 g / dl (2.2),
Alkaline phosphatase = 7.0 K-A units (5.
The value of 5) was obtained, indicating that the blood chemistry analysis material of the present invention is effective for quantifying components in blood.

Example 4 A liquid resin (4) was used in place of the (meth) acrylate liquid resin (1).
The same operation as in Example 1 was performed to obtain a blood chemistry analysis material.
Further, using this, the same measurement as in Example 1 was performed, and the respective component concentrations and reproducibility were obtained. as a result,
Glucose = 91 mg / dl (3.5), uric acid = 5.3
mg / dl (2.1), total cholesterol = 155 mg
/ Dl (6.5), calcium = 9.4 mg / dl
(4.1), albumin = 4.6 g / dl (1.9),
Alkaline phosphatase = 7.3 KA units (5.
The value of 3) was obtained, indicating that the blood chemistry analysis material of the present invention is effective for quantifying components in blood.

Example 5 As a (meth) acrylate monomer, instead of polyethylene glycol diacrylate (number average molecular weight = 508), 2,2-bis [4-
Except for using (acryloxy-polypropoxy) phenyl] propane (number average molecular weight = 560), the same operation as in Example 1 was performed to obtain a blood chemistry analysis material. Further, using this, the same measurement as in Example 1 was performed, and the respective component concentrations and reproducibility were obtained. As a result, glucose =
94 mg / dl (3.2), uric acid = 5.3 mg / dl
(2.6), total cholesterol = 150 mg / dl
(6.5), calcium = 9.7 mg / dl (4.
3), albumin = 4.7 g / dl (2.2), alkaline phosphatase = 7.1 KA unit (5.8) were obtained, and the blood chemistry analysis material of the present invention was used to determine the components in blood. Has been shown to be effective.

Example 6 A blood chemistry analysis material was obtained by performing the same operation as in Example 1 except that the electron beam was irradiated using MIN-EB instead of the area beam type electron beam. The irradiation dose of the electron beam was 5 Mrad. further,
Using this, the same measurement as in Example 1 was performed, and the respective component concentrations and reproducibility were obtained. As a result, glucose = 95 mg / dl (3.1) and uric acid = 5.1 mg / dl
(2.4), total cholesterol = 152 mg / dl
(6.3), calcium = 9.5 mg / dl (3.
7), albumin = 4.5 g / dl (2.1), alkaline phosphatase = 7.3 K-A units (5.6), and the blood chemistry analysis material of the present invention was used for the determination of components in blood. Has been shown to be effective.

Example 7 A blood chemistry analysis material was obtained by performing the same operation as in Example 1 except that γ-rays were irradiated instead of the electron beam. The irradiation dose of γ-ray was 10 KGy. Further, using this, the same measurement as in Example 1 was performed, and the respective component concentrations and reproducibility were obtained. As a result, glucose = 91 mg / dl (3.0), uric acid =
4.9 mg / dl (2.2), total cholesterol = 14
5 mg / dl (5.8), calcium = 9.2 mg / d
1 (3.0), albumin = 4.2 g / dl (2.
3), a value of alkaline phosphatase = 6.9 K-A units (5.7) was obtained, indicating that the blood chemistry analysis material of the present invention is effective for quantifying components in blood.

Example 8 The above-mentioned reagents for measuring glucose, uric acid, reagents for measuring total cholesterol, reagents for measuring calcium, reagents for measuring albumin and reagents for measuring alkaline phosphatase were added to 100 parts by weight of gelatin. 50 parts by weight were added and mixed. The obtained mixture was applied to a portion of a PET colorless transparent smooth sheet on which a blood cell separation section was to be formed as a reagent section, and dried until the water content became 10% by weight or less, as shown in FIG. 2 and FIG. A blood chemistry analysis material of various shapes was obtained. In addition, the thickness of the reagent part after hardening was 100 micrometers, and the diameter was 0.32 cm. Further, using this, the same measurement as in Example 1 was performed, and the respective component concentrations and reproducibility were obtained. As a result, glucose = 95 mg / dl (3.0), uric acid =
5.2 mg / dl (2.4), total cholesterol = 15
0 mg / dl (6.4), calcium = 9.6 mg / d
1 (3.9), albumin = 4.5 g / dl (1.
9), a value of alkaline phosphatase = 7.2 K-A units (5.3) was obtained, indicating that the blood chemistry analysis material of the present invention is effective for quantifying components in blood.

Example 9 A 200 μm thick PET colorless transparent smooth sheet was coated with an acetone-dichloroethane (1: 1) mixed solvent solution of cellulose acetate, and then dried. The blood cell separation part was prepared so as to have the shape shown in FIG. PET colorless transparent smooth sheet 1
The length of the side is 1.0 cm, and the blood cell separation part is 0.10 cm from the center of the sheet, 0.08 cm in length, and 0.10 in width.
cm, and the thickness was 100 μm. The same operation as in Example 8 was performed to form a reagent part, and a blood chemistry analysis material having a shape as shown in FIGS. 4 and 5 was obtained. In addition, the thickness of the reagent part after curing is 100 μm and the diameter is 0.32 cm.
Met. Further, using this, the same measurement as in Example 1 was performed, and the respective component concentrations and reproducibility were obtained. As a result, glucose = 96 mg / dl (3.2), uric acid = 5.3 mg / dl (2.6), and total cholesterol = 1
52 mg / dl (6.6), calcium = 9.7 mg /
dl (4.1), albumin = 4.6 g / dl (2.
1), a value of alkaline phosphatase = 7.3 K-A units (5.5) was obtained, indicating that the blood chemistry analysis material of the present invention is effective for quantification of components in blood.

[0069]

The characteristic of the blood chemistry analysis material of the present invention is that, unlike the conventional dry chemistry using a multilayer film type analysis material, blood cells are filtered and separated in the blood cell separation section while the blood spreads in the horizontal direction. That is, the plasma reaches the reagent part and a reaction occurs between the reagent and the test substance. Since the blood chemistry analysis material is configured as described above, a plurality of specific components can be measured at one time. Further, by making the hydrophilic carrier holding the reagent in the reagent section transparent, measurement by transmitted light is enabled. Further, the transparent hydrophilic carrier holding the reagent can be cured by irradiation with radiation, whereby the reagent portion can be easily obtained. Therefore, by using the blood chemistry analysis material of the present invention, it is possible to easily and accurately quantify a plurality of specific components in blood at once.

[Brief description of the drawings]

FIG. 1 is a cross-sectional view of a conventional blood chemistry analysis material.

FIG. 2 is a plan view of the blood chemistry analysis material of the present invention.

FIG. 3 is a cross-sectional view of the blood chemistry analysis material of the present invention.

FIG. 4 is a plan view of the blood chemistry analysis material of the present invention.

FIG. 5 is a cross-sectional view of the blood chemistry analysis material of the present invention.

[Explanation of symbols]

 A blood drop position and direction B reflection photometric direction a development layer b reflection layer c reagent layer d transparent support 1 support 2 blood cell separation section 3 reagent section 4 center of blood chemistry analysis material

Continuation of the front page (72) Inventor Dai Dai Matsui 2-163-2-206 Daisawa, Setagaya-ku, Tokyo

Claims (5)

[Claims] 1. A blood chemistry analysis material comprising a support and a blood cell separation section and an appropriate number of independent reagent sections arranged in a plane. 2. The apparatus according to claim 1, wherein an appropriate number of said blood cell separation sections and a number of said reagent sections corresponding to the number of said blood cell separation sections are arranged on said support in a plane. Item 7. A blood chemical analysis material according to item 1. 3. The blood chemistry analysis material according to claim 1, wherein the plurality of reagent parts are arranged at substantially the same position from a blood drop site. 4. The blood chemistry analysis material according to claim 1, wherein the blood cell separation section is made of a porous substance. 5. A reagent in which the reagent section reacts with a component in blood to exhibit a color having an absorbance at 200 to 900 nm or emits light at 200 to 900 nm, and a transparent hydrophilic carrier holding the reagent. The blood chemistry analysis material according to any one of claims 1 to 4, comprising: