JPH10206417A - Blood chemical analysis material and blood chemical analysis method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a blood chemical analysis material and an analysis method of components involved using the same which enable handy and highly accurate determination of a plurality of specific components in blood at a time. SOLUTION: A proper number of reagent parts 2 which each comprise a reagent that reacts with components in blood to form color with an absorbance of 200-900nm or emits light with a wavelength of 200-900nm and a transparent hydrophilic carrier for holding the reagent are provided horizontally on a light transmitting/non-water permeable support body 1 and a blood dripping part 4 placed thereabove is linked to the reagent parts 2 by a link part 3 having a blood cell separating function to laminate. Each of the reagent parts 2 is so arranged to be partially extended from the blood dripping part 4. The extended reagent parts 2 are measured by a transmission photometry to analyze the components.

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 and a method for analyzing the specific components using the material.

[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]

DISCLOSURE OF THE INVENTION The present invention provides a blood chemistry analysis material which can easily and accurately quantify a plurality of specific components in blood at once, and a method for analyzing the specific components using the same. The purpose is to do.

The present inventors laminated a support, an appropriate number of reagent portions, and blood dropping portions in that order, and formed the blood dropping portions such that a part of the surface of each of the reagent portions was exposed. And while providing a connection portion for connecting to the reagent portion inside thereof,
In particular, the present inventors have found that the use of a blood chemistry analysis material having a connection part having a blood cell separation function allows a specific component in blood to be easily and accurately determined at a time.

[0008]

That is, the present invention provides a support,
An appropriate number of reagent parts and blood dropping parts are blood chemistry analysis materials laminated in that order, and the blood dropping parts are formed so that at least a part of the surface of each of the reagent parts is exposed, and The gist of the present invention is a blood chemistry analysis material characterized in that a connection portion for connecting to the reagent portion is provided therein.

[0009] The blood chemistry analysis material of the present invention is characterized in that the connecting portion comprises a number corresponding to the reagent portion.

Further, the blood chemical analysis material of the present invention is characterized in that the connecting portion has a blood cell separating function.

Further, the blood chemistry analysis material of the present invention is characterized in that each of the above-mentioned reagent parts is arranged at substantially the same position from the blood dropping part.

Further, the blood chemistry analysis material of the present invention is characterized in that a part of each of the reagent parts is arranged to extend from the blood dropping part.

Further, in the blood chemistry analysis material of the present invention, the above-mentioned reagent part is a reagent which exhibits a color development having an absorbance at 200 to 900 nm or 200 to 90 when it reacts with a component in blood.
It is characterized by comprising a reagent emitting light of 0 nm and a transparent hydrophilic carrier holding the reagent.

Further, the present invention provides a method for analyzing a specific component in blood, characterized in that the exposed reagent portion of the blood chemistry analysis material is measured by a transmission photometric method through the support.

[0015]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS A structural example of the blood chemistry analysis material of the present invention will be described with reference to FIGS. FIG. 2 is a plan view showing a specific example of the blood chemistry analysis material of the present invention, and FIG.
FIG. 3 is a sectional view taken along line XY of FIG. 2. As shown in FIGS. 2 and 3, the blood chemistry analysis material of the present invention is placed on the support 1 from the center 5 of the blood chemistry analysis material, preferably at substantially the same position, and in a planar independent number. And a connecting portion 3 is provided inside the blood dropping portion 4 such that the blood dropping portion 4 and a part of the reagent portion 2 overlap with each other while being connected by the connecting portion 3. It is a structure arranged. The connecting portions 3 are provided independently of each other, but preferably have a number corresponding to the number of the reagent portions 2. The connecting portion 3 has a blood cell separating function, and it is preferable that a part of each reagent portion 2 extends from the blood dropping portion 4 as shown in the figure. The shape of the blood dropping part 4 is not particularly limited,
It is preferably a disk shape. In addition, the blood dropping section 4
Since it serves as a liquid pool at the time of blood filtration at the connecting portion 3 having a blood cell separating function, it is desirable to provide an edge 7 on the outer edge portion of the connecting portion 3 so as not to spill blood. Reference numeral 6 denotes an isolating member for isolating each reagent part 2 and also plays a role of supporting the blood dropping part 4. The blood chemistry analysis material of the present invention having such a configuration enables one blood chemistry analysis material to analyze a desired number of specific components in blood. The analysis of the specific component in the blood is performed by measuring the reagent part 2, a part of which is extended from the blood dropping part 4, by a transmission photometric method.

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

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 light absorbance at a wavelength in the range of 200 to 900 nm or 200
Those comprising a reagent emitting light of up to 900 nm and a transparent hydrophilic carrier holding the reagent are preferred. Specific components include, for example, chemical substances such as glucose, urea nitrogen, creatinine, uric acid, total cholesterol, high density lipoprotein (HDL) cholesterol, triglyceride, total bilirubin, calcium, inorganic phosphorus, total protein, albumin, ammonia and hemoglobin And γ-
Glutamyl transpeptidase (γ-GTP), glutamate oxaloacetate transaminase (GOT),
Glutamate pyruvate transaminase (GP
T), creatine phosphokinase, lactate dehydrogenase (LDH), alkaline phosphatase (ALP), amylase, leucine aminopeptidase and other enzymes. In the reagent section, these specific components react with the reagent, and depending on the amount of the components in plasma, 200 to
A color development having an absorbance at 800 nm, or 2
Since it emits light having a wavelength of from 00 to 800 nm, it is possible to quantify the components in the plasma by photometry.

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.

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

In the present invention, a radiation-curable curable liquid resin composition shown 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).
Is a (co) polymer obtained by polymerizing or copolymerizing 0 to 80% by weight of the polymerizable monomer (a-2) excluding the monomer (a-1) and the number average molecular weight. Is 10,0
It is a solventless liquid resin composition having a viscosity of 1 to 10,000 poise (50 ° C.) in the range of 00 to 200,000.

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)
Represents the monomer (a-1) and the polymerizable monomer (a-
2) is a solvent-free liquid resin obtained by copolymerizing the resin composition, by giving a suitable viscosity at the time of preparing the reagent portion by liquefying the resin composition,
When the reagent contains an enzyme or the like, it is also a component for stably presenting them in the reagent part. Further, it plays a role of imparting toughness and flexibility to the reagent part after curing.

The monomer (a-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- Pentyloxytetrapropylene glycol (meth) acrylate, methoxy polyethylene glycol (meth) acrylate, ethoxy polyethylene glycol (meth) acrylate, or phenoxytetraethylene glycol (meth) acrylate, phenoxyhexaethylene glycol (meth) acrylate, phenoxy polyethylene glycol (meth) ) Acrylate, phenoxytetrapropyleneglycol , And the like (meth) acrylate. Of these, the use of the monomer (a-1) having a polyoxyalkylene side chain in which m in the general formula (1) is 3 to 25, and preferably 4 to 22, makes it possible to obtain 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 with 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.

Examples of the radical polymerizable monomer having a carboxyl group in the molecule include, for example, maleic anhydride, maleic acid, fumaric acid, itaconic acid, citraconic acid, or an alkyl or alkenyl monoester thereof, and 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 and 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.

Examples of the radical polymerizable monomer having a hydroxyl group in the molecule include 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.

In the case where the resin composition used in the present invention is cured by irradiating an electron beam, a resin represented by the general formula (1)
Is a compound wherein R ¹ is a hydrogen atom.
In this case, the polymerizable monomer (a-2) used for the copolymerization
Is preferably one having no 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 monomers (a-1) and (a-2) in a solvent in the presence of a radical polymerization initiator, or a method of dissolving the monomers (a-1) and (a-2) Can be produced by radical polymerization using a method of dropping a mixture of the above. Examples of the radical polymerization initiator include benzoyl peroxide, t-butyl peroxide, cumene hydroperoxide, lauroyl peroxide, and other organic peroxides (eg, peroxides described in Taisei Co., Ltd., “Crosslinking Agent Handbook”, pages 520 to 535). Known compounds such as 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.

As the (meth) acrylate monomer (B), 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 portion of 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 coating the mixture on the support where the reagent portion is to be formed. Thereafter, it can be obtained by drying or curing. The reagent is generally prepared as a solution, and the amount added in this case is 0.1 to 1,000 parts by weight based on 100 parts by weight of the hydrophilic carrier or the curable liquid resin composition.
Desirably, parts by weight are used. Further, in order to improve the physical properties of the obtained reagent part, a compatibilizer and a surfactant may be added as necessary. These addition amounts are based on 100 parts by weight of the hydrophilic carrier or the curable liquid resin composition.
It is desirable that the content be 20 parts by weight or less, preferably 10 parts by weight or less.

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 section 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.

The blood dropping part 4 in FIGS. 2 and 3 is arranged on the reagent part 2 so that a part thereof overlaps. The part where the blood dropping part 4 and the reagent part 2 overlap is the connecting part 3 of the two, and when blood moves from the blood dropping part 4 to the reagent part 2, blood cells are separated, and only the plasma becomes the reagent part. 2, the connecting portion 2 has a blood cell separating function. The blood dropping section 4 can be provided by stacking on a separating member 6 for separating each reagent section 2. The material of the blood dropping portion 4 is not particularly limited, but may be the same material as the support 1. Further, the material of the separating member 6 is not particularly limited, but may be the same material as the support 1. When these reagent portions 2 are manufactured by a printing process, the material is suitable for the printing process and the Materials that can be isolated are appropriately selected. In addition, when the reagent part 2 is solid and has non-deformability, it may not be necessary. The blood dropping part 4 and the separating member 6 or the solid reagent part 2 may be simply laminated, but more preferably both are adhered by an adhesive. Examples of the adhesive to be used include various adhesives described in JP-A-62-138756, and others, for example, "Printing Engineering Handbook" edited by The Printing Society of Japan (Gihodo Shuppan Co., Ltd., 1983) 839.
Known adhesives described on pages pp. 853 can be used.
Furthermore, it is also possible to stick both using a double-sided tape.

The surface of the blood dropping portion can be subjected to hemolysis preventing treatment by a known method using a hemolytic agent such as di-2-ethylhexyl phthalate so as not to cause hemolysis when blood comes into contact. Further, in order to reduce the residual amount of blood in this portion, the surface of the blood dropping portion may be subjected to a water-repellent treatment by a known method using an organopolysiloxane or a fluoroalkyl compound.

It is desirable to use a porous material made of fibrous or non-fibrous material and having a hydrophilic surface for the connecting portion having a blood cell separating function. Examples of the fibrous porous substance include woven fabric, glass fiber, cellulose fiber, paper and the like, which are described in JP-B-61-61347. Examples of the non-fibrous porous substance include cellulose esters described in JP-B-53-21677 and U.S. Pat. No. 1,421,341, such as cellulose acetate, cellulose acetate / butyrate, and the like. Brush polymer made of cellulose acetate (generic name membrane filter),
Polyamides such as 6-nylon and 6,6-nylon, microporous materials such as polyethylene and polypropylene, small polymer particles, glass particles, diatomaceous earth, etc. have continuous voids bonded by hydrophilic or non-water-absorbing polymers Porous materials and the like can be mentioned.

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 pore distribution, and the like. 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 blood cell separating function can be easily provided by disposing a film cut into an appropriate size at a connecting portion between a blood dropping portion and a reagent portion. Can be. When the porous substance is not in the form of a film, a coating agent that forms a porous blood cell separation part, for example, a mixed solvent solution of cellulose acetate in acetone-dichloroethane (1: 1), silica gel, microcrystalline cellulose, A coating material in which 120 mesh glass beads are dispersed in a small amount of starch paste aqueous solution or gelatin solution is applied to a portion forming a connection portion between a blood dropping portion and a reagent portion and dried, and a porous substance is formed in a drying process. It is also possible to take a method such as When the hydrophilic carrier in the reagent section is made of the above-mentioned curable resin composition, the above-mentioned porous substance may not be used because the surface of the cured product becomes porous and has a blood cell separation function.

The thickness of the connecting part depends on the thickness of the blood dropping part arranged on the reagent part, but is desirably 5 to 500 μm, preferably 10 to 250 μm. If the thickness of the connecting portion is less than 5 μm, it is difficult to sufficiently perform filtration and separation of blood cells, which is not preferable. On the other hand, if the thickness of the connecting portion is greater than 500 μm, plasma is retained at the connecting portion, and a large amount of blood is required, which is not preferable.

The connection part can 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, it is possible to include various reagents for promoting the detection reaction in the reagent section, or for reducing or preventing interference and interference reactions, or a part of the reagents.

In the blood chemistry analysis material of the present invention, a light-permeable, water-impermeable film or the like may be laminated thereon, particularly for the purpose of protecting the reagent portion extending from the blood dropping portion. 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, a reagent portion optically exposed through a support is transmitted through a transmission photometric method. The measurement sensitivity can be improved as compared with dry chemistry using a conventional material.
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.

In the blood chemistry analysis material of the present invention, there is no problem even 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 FIG. 2, the length of one side is 0.5 to
Desirably, it is 2.0 cm.

[0055]

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 four 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-based liquid resin (1) In a 500 ml four-necked round-bottomed flask equipped with a stirrer, a nitrogen inlet tube, a temperature sensor and a condenser, methoxypolyethylene glycol acrylate (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 33
% 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 is 16,10
The viscosity at 0 and 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 (m = 9 in the above-mentioned general 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 Reagent for Glucose Measurement Each component was dissolved in a 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 1-2. 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 1-3. 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 1-4. 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 1-5. 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 each of the above-described reagents for measuring glucose, uric acid, reagents for measuring total cholesterol, and reagents for measuring calcium were added and mixed. The obtained mixture is applied to the four corners of a polyethylene terephthalate (PET) colorless and transparent smooth sheet having a thickness of 200 μm and a side length of 1.0 cm, and is irradiated with an electron beam using an area beam type electron beam irradiation apparatus. (DOSE = 5 Mrad) and cured to prepare a reagent portion having a shape as shown in FIGS. 2 and 3.
Note that the thickness of the reagent portion after curing is 100 μm and the diameter is 0.1 μm.
At 40 cm, the distance from the center of the sheet to the edge of the reagent section was 0.10 cm. Further, a circular PET colorless and transparent smooth sheet having a diameter of 0.20 cm was formed at the center of the sheet by using a hot-melt adhesive having a thickness of 100 mm.
It adhered so that it might be set to micrometers. 1-6. Preparation of blood dripping part: 100 μm thick with a 100 μm high edge on the outer periphery,
Four portions of a circular PET colorless transparent smooth sheet having a diameter of 0.60 cm were punched out, and a portion serving as a connecting portion having a diameter of 0.14 cm having a shape as shown in FIGS. 2 and 3 was provided. This was adhered to the above-mentioned circular sheet having a diameter of 0.20 cm using a hot-melt adhesive having a thermoadhesive property to prepare a blood dropping portion. 1-7. Preparation of a connection part having a blood cell separation function A cellulose fiber filter paper having an average pore diameter of 2.5 μm and a thickness of 100 μm is punched out into a circle having a diameter of 0.14 μm, and this is fitted into the above-described part serving as a connection part, thereby providing a blood cell separation function. Was prepared.

<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 each component concentration was calculated from a calibration curve between each component concentration and absorbance prepared in advance, glucose = 105 mg / dl, uric acid =
5.7 mg / dl, total cholesterol = 165 mg / d
l, calcium = 9.0 mg / dl. The same operation was repeated 10 times to check the reproducibility.
(%) Is glucose = 2.5, uric acid = 2.0, total cholesterol = 5.2, calcium = 3.2, and the measurement method using the blood chemistry analysis material of the present invention is sufficiently reliable. It was found to be a measurement method.

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 = 103 mg / dl (2.4), uric acid = 6.
0mg / dl (2.5), total cholesterol = 163m
g / dl (5.5), calcium = 9.0 mg / dl
(2.7), a value of albumin = 4.4 g / dl (2.0) was obtained, indicating that the blood chemistry analysis material of the present invention is effective for quantifying components in blood. 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 = 105 mg / dl (2.4), uric acid = 5.
6 mg / dl (2.3), total cholesterol = 167 m
g / dl (4.7), calcium = 9.2 mg / dl
The value of (3.1) 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 = 101 mg / dl (2.9), uric acid = 5.
8 mg / dl (1.6), total cholesterol = 170 m
g / dl (5.2), calcium = 8.8 mg / dl
The value of (3.3) 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 =
104 mg / dl (2.6), uric acid = 5.8 mg / dl
(2.1), total cholesterol = 165 mg / dl
(5.2), calcium = 9.1 mg / dl (3.5)
Was obtained, indicating that the blood chemistry analysis material of the present invention was effective in quantifying components in blood.

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 = 105 mg / dl (2.5), uric acid = 5.6 mg / d
1 (1.9), total cholesterol = 167 mg / dl
(5.0), calcium = 8.9 mg / dl (2.9)
Was obtained, indicating that the blood chemistry analysis material of the present invention was effective in quantifying components in blood.

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 = 101 mg / dl (2.4), uric acid =
5.4 mg / dl (1.7), total cholesterol = 16
0 mg / dl (4.5), calcium = 8.6 mg / d
A value of l (2.2) 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 used for 100 parts by weight of gelatin. 50 parts by weight were added and mixed. The obtained mixture was applied to the four corners of a polyethylene terephthalate (PET) colorless transparent smooth sheet having a thickness of 200 μm and a side length of 1.0 cm, and dried until the water content became 10% by weight or less. 2 and FIG. 3 were prepared, and then the same operation as in Example 1 was performed to obtain a blood chemistry analysis material. In addition, using this,
The same measurement as in Example 1 was performed, and the respective component concentrations and reproducibility were obtained. As a result, glucose = 105 mg /
dl (2.4), uric acid = 5.7 mg / dl (1.9),
The values of total cholesterol = 165 mg / dl (5.1) and calcium = 9.0 mg / dl (3.1) were obtained, indicating that the blood chemistry analysis material of the present invention is effective for quantifying components in blood. Indicated.

[0069]

Since the blood chemistry analysis material of the present invention has the above structure, it is possible to measure a plurality of specific components at one time. In addition, by making the hydrophilic carrier holding the reagent in the reagent section transparent, each reagent section extends from the blood dropping section, so that measurement by transmitted light is possible. 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, a desired number of specific components in blood can be easily and accurately quantified at a time.

[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.

[Explanation of symbols]

 A Blood Dropping Position and Direction B Reflection Photometric Direction a Spreading Layer b Reflective Layer c Reagent Layer d Transparent Support 1 Support 2 Reagent Section 3 Connecting Section 4 Blood Dropping Section 5 Center of Blood Chemical Analysis Material 6 Separating Member 7 Edge

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Dai Dai Matsui 2- 16-3-206 Daisawa, Setagaya-ku, Tokyo

Claims (7)

[Claims] 1. A blood chemistry analysis material comprising a support, an appropriate number of reagent parts, and a blood dropping part laminated in that order, wherein the blood dropping part exposes at least a part of the surface of each of the reagent parts. A blood chemistry analysis material characterized in that it is formed as described above, and is provided therein with a connecting portion for connecting to the reagent portion. 2. The blood chemistry analysis material according to claim 1, wherein the number of the connecting portions is equal to the number of the reagent portions. 3. The blood chemistry analysis material according to claim 1, wherein the connecting portion has a blood cell separating function. 4. The blood chemistry analysis material according to claim 1, wherein each of said reagent portions is arranged at substantially the same position from a blood drop site. 5. The method according to claim 1, wherein a part of each of the reagent parts is arranged to extend from the blood dropping part.
5. The blood chemical analysis material according to any one of 4. 6. The reagent, wherein the reagent part 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 chemical analysis material according to any one of claims 1 to 5, comprising: 7. An analysis of a specific component in blood, wherein the exposed reagent portion of the blood chemistry analysis material according to claim 1 is measured by a transmission photometric method through the support. Method.