JPH03242539A - Test device and manufacture thereof - Google Patents

Abstract

PURPOSE:To detect a specified substance such as glucose and occult blood accurately in an object to be inspected by setting 10-700mum for a thickness of an oxidizer layer of a test device in which a reagent layer carrying a reagent and an oxidizer layer containing an oxidizer are provided on a support. CONSTITUTION:A test device 1 has a reagent layer 4 and an oxidizer layer 5 with a thickness of 10-700mum laminated in the order on one side of a support 3. The support 3 is composed of cellulose ester. polypropylene and the like. The reagent layer 4 employs a non-fibrous porous filter paper, fibrous porous cotton, silk, wool or the like as carrier. The reagent herein used is an enzyme of hydroperoxide, glucose oxidase or the like that forms a color when being oxidized. The oxidizer layer 5 is formed at a thickness of 10-700mum using a fine powder filter paper. The oxidizer herein used is ferric chloride, cuprous chloride or the like as oxygen acid of HClO, NaClO or the like and a salt and a metal salt thereof.

Description

[Detailed Description of the Invention] A test device for detecting urine, feces, and blood in liquid <Industrial Application Field> The present invention provides a test device for detecting a specific substance in a specimen,
In particular, the present invention relates to a test device for detecting glucose in urine and blood, and for detecting blood in urine, feces, and liquid, and a method for manufacturing the same.

<Prior Art> In recent years, rapid diagnostic agents have become extremely important in medical practice and clinical trials, and test devices carrying these rapid diagnostic agents have been developed. In this test device, an absorbent carrier such as paper is impregnated with a drug necessary for a detection reaction, and when this is immersed in a body sample, a color reaction occurs when the substance to be detected is present. arises,
It is possible to recognize the existence of the substance.

Such test devices (test strips) include those for detecting glucose in urine and blood, for example.

These principles are as follows.

When detecting glucose in body fluids, for example, glucose is oxidized to gluconic acid by glucose oxidase (GOD), and at the same time oxygen in the air is reduced to hydrogen peroxide. This hydrogen peroxide is produced by peroxidase (P
OD) or peroxidase-active substances, the reagent (oxidized color indicator) is oxidized to the corresponding dye, the concentration of the color being a measure of the amount of glucose present.

Furthermore, when detecting blood, active oxygen is generated from peroxide contained in the test strip or the like in advance due to the peroxidase-like activity of hemoglobin, which oxidizes the oxidized coloring indicator and causes it to develop color.

By the way, recent processed foods, soft drinks, etc. contain vitamin C.
It contains a large amount of ascorbic acid, and vitamins are also taken in large quantities for health management reasons.

Therefore, a large amount of ascorbic acid is contained in the blood, and excess ascorbic acid is contained in the urine and excreted from the body. For this reason, when testing urine or blood using the test device described above, ascorbic acid has strong reducing properties, so active oxygen generated by peroxidase or peroxidase-active substances is consumed in the redox reaction with ascorbic acid. The problem has arisen that the reaction of the oxidized color indicator is inhibited and sufficient color development cannot be obtained.

In order to solve this problem, a technique has been disclosed in which an effective amount of oxidizing agent is added to the test agent to eliminate the influence of reducing substances such as ascorbic acid (Japanese Patent Laid-Open No. 60-86
However, with this test device, in addition to being affected by oxygen and moisture in the air, the reagent (oxidized coloring indicator) and oxidizing agent are immersed in the same test agent, so it is difficult to test. There was a drawback that the reagents were oxidized during storage before use, causing coloration.

In addition, in order to solve the above-mentioned problems, we impregnated and coated membrane base materials such as mesh-like or multiple pinhole nonwoven fabrics, porous membranes, glass fibers, semipermeable membranes, and filter paper with an oxidizing agent. Alternatively, a technique has been disclosed in which the influence of reducing substances such as ascorbic acid is eliminated by providing an attached oxidizing agent layer, and the stability of the test device over time is ensured (Japanese Patent Laid-Open No. 60-224063). ) However, since this test device uses a membrane base material with a certain thickness as the oxidizing agent layer, the transparency of the oxidizing agent layer decreases, and it is difficult to set the fiber density and thickness accurately and uniformly. Since it is difficult to do so, there are problems such as color unevenness occurring during coloring, and this has the disadvantage that it is difficult to accurately observe the coloring state.

<Problems to be Solved by the Invention> The present invention has been made in view of the above-mentioned drawbacks of the prior art, and its purpose is to prevent changes over time during storage and to accurately observe the color development of the reagent layer. The object of the present invention is to provide a test device that enables measurement, and a method for manufacturing the same.

<Means for Solving the Problems> Such objects are achieved by the following inventions (1) to (5).

(1) A test device having a reagent layer carrying a reagent and an oxidizing agent layer containing an oxidizing agent on a support, characterized in that the thickness of the oxidizing agent layer is 10 to 700 mm. test equipment.

(2) The test device according to (1) above, wherein the oxidizing agent layer is composed of a fine powder carrier supporting an oxidizing agent.

(3) The test device according to (1) or (2) above, wherein a binder layer containing a binder is provided on the oxidizing agent layer.

(4) The binder may include gelatin, gum arabic, polyvinylpyrrolidone, polyvinyl alcohol, agarose,
Sodium polyvinylbenzenesulfonate, polyurethane, polyvinyl propionate, polyethylene glycol, acrylate, polyacrylamide, poly(hydroxyethyl methacrylate) poly(hydroxyethyl acrylate) selected from the group consisting of carboxymethyl cellulose, methyl cellulose, hydroxypropyl cellulose The test device according to (3) above, which is at least one type.

(5) A test characterized in that a reagent layer carrying a reagent is formed on a support, and an oxidizing agent layer is formed by spraying a fine powder carrier carrying an oxidizing agent onto the reagent layer. Method of manufacturing ingredients.

<Function> In the test device of the present invention, by providing the oxidizing agent layer, it is possible to eliminate the influence of reducing substances such as ascorbic acid and to prevent changes over time during storage.

In this case, since the thickness of the oxidizing agent layer is 10 to 700 mm, transparency of the oxidizing agent layer is ensured, and color changes in the reagent layer can be observed and measured accurately.

In addition, in the test device of the present invention, when the oxidizing agent layer is composed of a fine powder carrier supporting the oxidizing agent, the amount of the oxidizing agent supported and the thickness of the oxidizing agent layer should be set uniformly. This makes it possible to prevent color unevenness during color development and ensure the stability of the reagent over time.

Furthermore, in the test device of the present invention, when a binder layer containing a binder is provided on the oxidizing agent layer, the influence of oxygen and moisture in the air can be reduced, and the test device is more stable over time. gender is ensured.

In addition, in the method for manufacturing the test device of the present invention, the oxidizing agent layer is formed by spraying a powder carrier carrying an oxidizing agent onto the reagent layer, so the thickness of the oxidizing agent layer can be easily adjusted. It can be set uniformly, and uneven coloring can therefore be prevented.

<Example> Hereinafter, a preferred example of the test device of the present invention shown in the accompanying drawings will be described in detail.

The test device of the present invention is preferably one that measures hydroperoxide or hydrogen peroxide or hydroperoxide generated by a pre-reaction by a peroxidase or peroxidase-like reaction, or a test device that measures peroxidase or a peroxidase-active substance (for example, hemoglobin).
It is used to measure.

An example of the configuration of such a test device is shown in FIG.

As shown in the figure, the test device 1 has a reagent layer 4 and an oxidizing agent layer 5 laminated in this order on one side of a support 3.

The support 3 has a plate shape with a thickness of about 20 to 200 mm, for example, and is preferably made of a light-transmitting material, that is, a transparent or translucent material.

Specific constituent materials of the support 3 include, for example, polyethylene terephthalate, cellulose ester (cellulose diacetate, cellulose triacetate, cellulose acetate propionate, etc.), polycarbonate of bisphenol A, polymethyl methacrylate, polyvinyl chloride, polypropylene, polystyrene. , various resins such as polyvinyl alcohol, or glass.

Alternatively, sheets made of two or more of the above materials may be laminated.

The reagent layer 4 is composed of a carrier supporting a reagent to be described later.

The carrier is preferably made of a non-fibrous or fibrous porous material.

Typical examples of non-fibrous porous materials include filter paper and membrane filters, as well as dispersions of porous materials such as diatomaceous earth and microcrystalline materials dispersed in a binder, and microspherical beads of glass or resin that are bonded to each other. Examples include porous aggregates bonded at points.

Furthermore, examples of the fibrous porous material include woven or knitted fabrics, nonwoven fabrics, short fiber aggregates, and the like.

Examples of the fiber material include natural fibers such as cotton, silk, and wool, and resins such as glass and nylon.

The main agents constituting the reagent include oxidized color indicators (coloring agents) that develop color when oxidized, hydroperoxides, and enzymes such as glucose oxidase and peroxidase, and these are used as appropriate depending on the substance to be detected.

Oxidation color indicators are used as a measurement system for oxygen generated by peroxidase or peroxidase-like reactions, reactions that produce oxidized condensates (phenol and 4-aminoantipyrine, etc.), methanol is oxidized to formaldehyde, and acetylacetone and condensates are produced. It is involved in the reaction in which iodide such as potassium iodide is oxidized to produce iodine, etc.

Examples of the oxidation color indicator include benzidine compounds, heterocyclic azines, phenols, and guaiac resin components. Specifically, oh-lysine, m-tolidine, benzidine, 3.3°, 5,5°-tetra(lower alkyl)benzidine, 0-methylbenzidine, 2,7-guamitsufluorene, or various combinations of these A mixture containing in proportions, 4.4'-diaminodiphenyl, 0-phenylenediamine, m-phenylenediamine, p-phenylenediamine, 2.3-tolylenediamine, 2.4-tolylenediamine, 2.5-tolylenediamine Amines, various substituted phenylenediamines such as 2.6-)-lylenediamine, amorphous gallic acid, gallic acid, phenols such as phloroglucinol, hydroquinone, leucoindophenol, guaiacol, pyridine derivatives, substituted azines, leuco There are malachite green, etc. In particular, 0-)
-Lysine is preferably used.

Hydroperoxides used in the present invention include bis[4-(a-hydroperoxyisopropyl)benzyl coether, 2,5-dimethylhexane-2,5-dihydroperoxide, cumene hydroperoxide, 2.5-dimethylhexane-2,5-dihydroperoxide,
-dimethylhexanone-2,5-dihydroperoxide, diisopropylbenzene hydroperoxide, t-butyl hydroperoxide, p-menthane hydroperoxide, 4-methylphenylisopropyl hydroperoxide and the like.

Moreover, various additives as described below can be added to the reagent layer 4 as necessary.

pH adjustment (pH = 4 to 8, preferably 5 to 7) when the reagent layer 4 and the specimen come into contact (for example, by immersing the test device in the specimen)
) Examples of the buffering agent include citric acid-sodium citrate, tartaric acid-sodium tartrate, malic acid-borax, potassium hydrogen phthalate-dipotassium phthalate, sodium hydrogen succinate-disodium succinate, and the like.

Further, when the reagent layer 4 and the specimen come into contact, wetting agents used to uniformly wet (penetrate) the specimen include, for example, polyvinylpyrrolidone, sodium lauryl sulfate,
Examples include alkyl sulfates such as sodium dodecyl sulfate and sodium tetradecyl sulfate, alkyl benzene sulfonates such as sodium dodecylbenzenesulfonate, and dialkyl sulfosuccinates such as sodium dioctyl sulfosuccinate and sodium diheptyl sulfosuccinate. Among these, especially polyvinylpyrrolidone,
It has the effect of accelerating the penetration of the specimen and has the advantage of increasing the speed of color development.

Further, in order to improve the coloring stability after coloring, a copolymer of methyl vinyl ether and maleic anhydride, or a half ethyl ester thereof, etc. may be added.

In addition, as a stabilizer for preventing changes over time during storage of test devices, Japanese Patent Publication No. 56-43238 and Japanese Patent Application No. 01-01
238148 (especially 2-mercaptobenzimidazole) and the like can also be added.

Furthermore, a drug that enhances peroxidase activity in hemoglobin may be added.

Typical enhancers include quinoline and its derivatives,
For example, quinine, cinchonine, 6-medoxyquinoline,
Quinaldine, 8-amino-6-medoxyquinoline, 2-
Quinoli tool, isoquinoline, benzo(f)quinoline,
Examples include 3-aminoquinoline. The presence of these enhancers usually accelerates the oxidation reaction rate and the color intensity of the oxidized chromogen, increasing the sensitivity of the test device.

Furthermore, a thickening agent may be added to prevent the drug from flowing out from the carrier of the reagent layer 4). Typical thickening agents include polyvinyl alcohol, polyvinylpyrrolidone,
Polyethylene di-1-fucol, acrylate, polyacrylamide, poly(hydroxyethyl methacrylate)
(Hydroxyethyl acrylate) Examples include polymers such as carboxymethyl cellulose, gelatin, and gum arabic.

The above reagents and additives can be supported on the carrier by impregnating the carrier with a liquid containing the reagent etc. (for example, by immersing the carrier in an immersion liquid), and then drying it.

When there are two or more types of acid bodies, impregnation and drying are repeated.

Note that such a reagent layer 4 is fixed to the surface of the support 3 using, for example, double-sided adhesive tape (not shown), adhesive, or the like.

In addition, the reagent layer 4 contains liquid containing reagents, etc.
Kuha Tsuchibu Ikkigu Koguchihiro 1 Ti base ^h, r-<+
(7) is also acceptable.

In this case, the reagent layer 4 contains a predetermined reagent or the like in a hydrophilic binder such as gelatin, polyvinyl alcohol, polyvinylpyrrolidone, agarose, sodium polyvinylbenzenesulfonate, polyurethane, or polyvinylpropionate. Composed of compositions.

The thickness (dry film thickness) of such reagent layer 4 is not particularly limited, but is approximately 0.5' to 2cm, particularly 0.6 to 1.0m.
About m is preferable.

As shown in FIG. 1, an oxidizing agent layer 5 containing an oxidizing agent is provided on the reagent layer 4.

The oxidizing agent layer 5 is preferably one in which an oxidizing agent, which will be described later, is supported on a fine powder carrier.

As the micropowder carrier, it is preferable to use powdered filter paper, microbeads of glass, resin, etc., and particularly preferred is micropowder filter paper having water absorption properties.

The reason is that by using such fine powder, the density (concentration) of the oxidizing agent can be made uniform, and the oxidizing agent layer 5 can be easily formed to a desired thickness.
Moreover, since the thickness can be made uniform, color unevenness during coloring can be prevented.

The diameter (average) of such a fine powder carrier is 20
About 200 μs, especially about 20 to 50 μs is preferable.

The oxidizing agent is one that can eliminate the influence of reducing substances such as ascorbic acid present in the specimen, and preferably oxyacids or salts thereof and metal salts can be used.

As the oxygen acid or its salt, MXOlMXO, MX
O,, M3H2XO6 (where M is an atom that can be a monovalent cation, such as Na, H, etc., and X is a halogen atom, such as I, Br, CI2, etc.)
etc.

Specific examples include HCβO, NaCβ0, KCj20
．． HBr0. NaBr0. KBrOlHIO,Nal0
．． KIO, HC (203NaCQO, KCffO
3, HBr0 3NaBrO3, K B r O3,
HIO,, N a I 03KIO,, HCQO
4, N a CQ O4, KCl20°HBr0. , N
a B r O4, K B r O4, HIO4N
al0< KIO4, Nas H2I06 to 3
There are H2I06 etc. Among these, MXO is preferred, MXO4 is more preferred, and NaIO4 or HIO4 is most preferred.

Metal salts include ferric chloride, cuprous chloride, copper sulfate,
Examples include copper acetate, mercury acetate, bismuth acetate, and lead acetate.

Such an oxidizing agent is used in an amount effective to prevent reaction inhibition of the oxidized color indicator by reducing substances such as ascorbic acid present in the specimen.
That is, the content of the oxidizing agent in the oxidizing agent layer 5 depends on the amount of the oxidized color indicator contained in the reagent layer 4 and the amount of reducing substance present in the specimen, but usually the content of the oxidizing agent in the test device 10XIO-' to 40X 10 mol/cm'' per unit area,
Preferably 20xlO-' to 30 per unit area of test device
It is preferable to set it to about xlO-'mol/cm2.

Moreover, the following additives can be added to the oxidizing agent M5 as necessary.

Main additives include binders, binder solubilizers, spray solvents, and the like.

As the binder, those mentioned below can be used, and the content of the binder in the oxidizing agent layer 5 is preferably about 0.05 to 2 wt%.

Further, examples of the solubilizing agent for the binder include alcoholic solvents such as ethanol.

Further, as a spray solvent, a hydrocarbon solvent such as ethyl acetate or chloroform can be used.

In the present invention, for example, a layer containing an oxidizing agent in a binder may be formed on the reagent H4 as the oxidizing agent layer 5 without using the above-mentioned fine powder carrier. Such oxidizer N5
can be formed, for example, by the method (2) described below.

By providing such an oxidizing agent layer 5, the specimen comes into contact with the oxidizing agent, the reducing properties of reducing substances such as ascorbic acid present in the specimen are lost, and the activity generated by peroxidase or peroxidase-active substances is reduced. Oxygen is no longer consumed by the reducing substance, and the coloring reaction of the oxidized coloring indicator is properly carried out.

In particular, since the oxidizing agent layer 5 containing the oxidizing agent is provided separately from the reagent layer 4, the reagent and the oxidizing agent do not come into contact with each other, or even if they do, only a portion of the contact occurs.
There is no change over time during storage, especially when the reagent is oxidized by an oxidizing agent and develops color.

Note that the oxidizing agent N5 is applied to the surface side of the reagent layer 4 (support 3
It is preferable to provide it on the opposite side).

Examples of methods for forming such an oxidizing agent layer 5 include the following.

(2) A liquid containing (dispersing) an oxidizing agent is coated on the surface of the reagent layer 4, preferably by spraying, and this is dried. In this case, a part of the solvent (dispersion medium) of the t-cover sprayed on the surface of the reagent layer penetrates into the reagent layer 4, but the oxidizing agent remains on the surface of the reagent layer and forms a layer by drying. do.

In this method, compared to method (3) below, the boundary between the reagent layer 4 and the oxidizer layer 5 is not necessarily clear, and the reagent and oxidizer may be present at the boundary. Since the amount is small compared to the entire reagent, the effect of the present invention of improving storage stability is fully exhibited.

(2) Impregnating a water-absorbing fine powder carrier (for example, powder filter paper) with a liquid containing an oxidizing agent, then applying the water-absorbing fine powder carrier to the surface of the reagent layer 4, preferably by spraying, and then drying it. . As a result, an oxidizing agent H5 is formed on the reagent layer 4, in which the oxidizing agent is supported on a water-absorbing fine powder carrier.

The thickness of such an oxidizing agent layer 5 is preferably about 10 to 700 μs. In particular, when the oxidizing agent layer is formed by the method (2) above, the thickness is preferably 10 to 500 mm, more preferably 50 to 200 mm. When using the method (2) above, the thickness is preferably 200 to 700 mm, more preferably 300 to 500 mm.

If the thickness of the oxidizing agent layer 5 is too thin, the ability to support the oxidizing agent will decrease, and the ability to eliminate the influence of ascorbic acid and the like (ascorbic acid resistance) will decrease.

Moreover, if the thickness of the oxidizing agent layer 5 is too thick, the transparency of the oxidizing agent layer will decrease, making it difficult to observe or measure the coloration of the reagent N4.

FIG. 2 shows another example of the configuration of the test device of the present invention. The test device 2 shown in the figure has a binder layer 6 containing a binder provided on an oxidizing agent layer 5.

This binder layer 6 serves to stabilize the layers below it, particularly the oxidizing agent layer 5, against the support 3, or to reduce the effects of oxygen and moisture in the air.

In this test device 2, the structures of the support 3, reagent layer 4, oxidizing agent layer 5, etc. are the same as those in the test device 1 described above.

Binders include gelatin, gum arabic, polyvinylpyrrolidone, polyvinyl alcohol, agarose,
Sodium polyvinylbenzenesulfonate, polyurethane, polyvinyl propionate, polyethylene glycol, acrylate, polyacrylamide, poly(hydroxyethyl methacrylate), poly(hydroxyethyl acrylate), polymers such as carboxymethylcellulose, methylcellulose, hydroxypropylcellulose It is preferable to use one or more of these.

Moreover, an organic solvent such as ethanol can be added to the binder N6 as necessary, for example, as a solubilizing agent for the binder.

By providing the binder layer 6 as described in No. 2, it is possible to prevent the oxidizing agent layer 4, especially the oxidizing agent layer 4 from collapsing due to the fine powder carrier, when cutting the test paper (a laminate of M4.5 and M6). As a result, uneven coloring is prevented from occurring.

The binder layer 6 is preferably formed by applying a liquid containing the binder to the surface of the oxidizing agent layer 5 and drying it.

In this case, the application of the liquid containing the binder is preferably carried out by spraying the liquid.

In addition, in such a formation method, there is no problem even if the liquid containing the binder permeates into the oxidizing agent layer 5 and some of the binding agent is present in the oxidizing agent layer 5.

The thickness of such a binder layer 6 is not particularly limited, but
It is preferably about 10-100 houses, especially about 40-60 houses.

Although the illustrated test devices 1 and 2 have been described above, the test device of the present invention is not limited to these.
For example, a device having two or more reagent layers carrying different reagents, or a light antireflection layer, a blood cell removal layer, a sample spreading layer, a sample absorption layer, a sample amount layer, etc. between or on each layer 4.5.6. It may also be provided with other layers such as an adjustment layer.

The test device of the present invention is applied to detect substances such as glucose, uric acid, cholesterol, triglyceride (neutral fat), free fatty acids, hemoglobin, myoglobin, and white blood cells.

<Experiment example> Experimental examples of the present invention will be described below.

(Example 1 of the present invention) Solutions 1.2 and 3 having the compositions shown below were prepared, and filter paper with a thickness of about 1 mm (N manufactured by Toyosha Co., Ltd.) was prepared.

1650) was first impregnated with Solution 1 and dried (40°C, 20°C).
minutes), then impregnated with solution 2 and dried (40℃, 50 minutes)
After that, it was impregnated with Solution 3 and dried (40°C, 10 minutes).

Next, spray liquid 4 having the composition shown below on one side of the filter paper.
Spray and dry at 40"C for 20 minutes to a thickness of 200"
An oxidizing agent layer of μ was formed.

Thereafter, this was cut into a size of 5 mm x 5 mm and adhered to a polystyrene sheet support using double-sided adhesive tape to obtain a test device for detecting occult blood in urine.

In addition, this test device has a concentration of 0.03 to 0.75 per 1 dj of urine.
It produces a blue color that allows the measurement of mg of hemoglobin.

Straight from the farm Bis[4-(α-hydroperoxyisopropyl)benzylconityl 777mgP-)luenesulfonyl-N-diethylamide 5゜Sodium dioctylsulfosuccinate
1.5g 6-medoxyquinoline
0.6ml! ethanol
100m1! Hama J1 Dan Acrylamide Polyethylene Glycol (Average Molecular Weight = 3,000) Citrate Sodium Citrate Saponin Ethylenediamine Tetraacetic Acid Disodium 0 g 0 g g g 00mg Mu Distilled Water Raku J1 Dan 0-Tolidine (Oxidation Color Indicator) Acetone Tamani Beggar: Ko L1 Sodium iodate (oxidizing agent) Polyvinylpyrrolidone ethanol ethyl acetate 0mg 00ml 1.2g 00mg 0.5g 0.4g 3 ml! 7 m2 (Example 2 of the present invention) 10 g of powder filter paper with an average particle size of 40 p was added to a 10% aqueous sodium iodate solution and then dried to obtain 10.5 g of powder filter paper supporting sodium iodate. 0.5 g of this powder filter paper and 0.4 g of polyvinylpyrrolidone were added to 3 ml of ethanol and 17 m2 of ethyl acetate.
Spray liquid 5 was obtained. This spray solution 5 was sprayed uniformly onto one side of a filter paper carrying the same reagent as above, and then dried at 40°C for 30 minutes to a thickness of about 300 μm.
An oxidizing agent layer of m was formed.

Other conditions were the same as in Example 1 of the present invention to obtain a test device for detecting occult blood in urine.

(Example 3 of the present invention) Solution 6 having the composition shown below was sprayed onto the oxidizing agent layer of the test device of Example 2 of the present invention, and then this was dried at 40°C for 30 minutes to form a bond with a thickness of approximately 50 units. A coating layer was formed.

Other conditions were the same as in Example 2 to obtain a test device for detecting occult blood in urine.

Accident 1 Polyvinylpyrrolidone 0.4g ethanol 3mj ethyl acetate
17 m2 (Comparative Example 1) The spray liquid 4 was applied to a thickness of about 1000 μm as an oxidizing agent layer.
Example 1 of the present invention except that an oxidizing agent film obtained by impregnating and drying the nonwoven fabric of No.
The same test device was used.

(Comparative Example 2) A test device similar to Inventive Example 1 was used except that the thickness of the oxidizing agent layer was set to about 1000 μm.

(Comparative Example 3) A test device was prepared in the same manner as in Inventive Example 1 except that the oxidizing agent layer and the binder layer were not provided.

(Comparative Example 4) A test device was prepared in the same manner as in Comparative Example 3, except that 0.05 mg of sodium iodate was supported as an oxidizing agent in the reagent layer (filter paper).

For each of the above test devices, hemoglobin was detected using urine whose hemoglobin concentration and ascorbic acid (ASA) concentration were adjusted as shown in Table 1, and the ascorbic acid (ASA) resistance of each test device was investigated.

The results are shown in Table 1 below.

The hemoglobin concentration was measured by measuring the color intensity of the test paper using a reflection absorbance meter.

Further, the hemoglobin concentrations in Table 1 are as follows.

...Omg/dj + -=0. 03mg/dj 2 +---0, 15mg/dffi 3 +-0, 75mg/dj 4 +-= 1, 50mg/cN Table 1 As shown in Table 1, the test devices of Examples 1.2 and 3 of the present invention Both have excellent ascorbic acid tolerance, and the hemoglobin concentration in urine can be accurately measured. In particular, in Invention Examples 2 and 3, no color unevenness occurred at all.

On the other hand, since the test device of Comparative Example 1 uses a nonwoven fabric for the oxidizing agent layer, there is a lot of uneven coloring, and since the oxidizing agent layer is as thick as 1000 μm, it lacks transparency and discoloration. It was difficult to observe accurately.

In addition, since the test device of Comparative Example 2 uses the oxidizing agent itself as the oxidizing agent layer, it is slightly better than Comparative Example 1 in terms of color unevenness, but because the oxidizing agent layer is as thick as 1000 μs, , it lacked transparency and it was difficult to observe coloration accurately.

Furthermore, since the test device of Comparative Example 3 does not contain an oxidizing agent, it has poor resistance to ascorbic acid, and the measured value of hemoglobin concentration is lower than the actual concentration.

Furthermore, since the test device of Comparative Example 4 contains an oxidizing agent in the reagent layer, it is excessively colored, and therefore the measured value of hemoglobin concentration is larger than the actual concentration.

In addition, since the test device of Inventive Example 3 has a binder layer,
Compared to the test device of Example 2 of the present invention, there was less uneven coloration.

Next, each test device of Examples 1 to 3 of the present invention and Comparative Examples 3 and 4 (
(Unused) was stored for 2 weeks in an environment of 60° C. and 20-30% RH, and changes over time (preservability) were examined. Then, hemoglobin was detected in the same manner as described above for each test device after storage.

The results are shown in Table 2.

The test devices of Invention Examples 1.2 and 3 did not show any color development or discoloration even after two weeks, and had excellent storage stability.

On the other hand, the test device of Comparative Example 3 has no problems with storage stability, but the test device of Comparative Example 4 starts to develop color after just one day, and completely develops color after two weeks, making it unusable. Met.

Next, test devices for detecting glucose in urine were prepared in the same manner as in Examples 1 to 3 of the present invention and Comparative Examples 1 to 4, except that the composition of the reagent was changed.

In this case, the filter paper (reagent layer) was sequentially immersed in molten acids 7 and 8 having the composition shown below, and dried at 40° C. for 60 minutes and 10 minutes, respectively.

Peroxidase 3.0g Peroxidase 100mg Tartrazine 150mg 10% Polyvinylpyrrolidone 30ml (K2O) Water volume (5%) Gantrez AN139 (Methylvinyl-
70+nj coelectrochemical combination of ether and maleic anhydride) water, g/? ! Citrate buffer i (1 (pH 5, 5) 50m
j Sodium metaperiodate (in the case of Comparative Example 2) 150 mg Raku' (dan0-tolidine 1.0 g) For each of these test devices, ascorbic acid When the resistance was investigated, the same results as in Table 3 above were obtained.

Furthermore, when the preservability was investigated in the same manner as above, the same results as in Table 2 above were obtained.

<Effects of the Invention> As described above, according to the present invention, it is possible to eliminate the influence of reducing substances such as ascorbic acid, prevent changes over time during storage, and furthermore, it is possible to accurately observe coloration. , can be measured. Therefore, specific substances such as glucose and occult blood in the sample can be detected more accurately.

acetone 00mj

[Brief explanation of drawings]

FIG. 1 and FIG. 2 are cross-sectional side views each showing an example of the configuration of the test device of the present invention. Explanation of symbols 1.2 Test device 3 Support 4 Reagent layer 5 Oxidizing agent layer 6 Binder layer Applicant: Terumo Corporation

Claims (5)

[Claims] (1) A test device having a reagent layer supporting a reagent and an oxidizing agent layer containing an oxidizing agent on a support, characterized in that the thickness of the oxidizing agent layer is 10 to 700 μm. test equipment. (2) The test device according to claim 1, wherein the oxidizing agent layer is composed of a fine powder carrier supporting an oxidizing agent. (3) The test device according to claim 1 or 2, further comprising a binder layer containing a binder provided on the oxidizing agent layer. (4) The binder may include gelatin, gum arabic, polyvinylpyrrolidone, polyvinyl alcohol, agarose,
Selected from the group consisting of sodium polyvinylbenzenesulfonate, polyurethane, polyvinyl propionate, polyethylene glycol, acrylate, polyacrylamide, poly(hydroxyethyl methacrylate), poly(hydroxyethyl acrylate), carboxymethylcellulose, methylcellulose, hydroxypropylcellulose The test device according to claim 3, wherein the test device is at least one type of. (5) A test characterized in that a reagent layer carrying a reagent is formed on a support, and an oxidizing agent layer is formed by spraying a fine powder carrier carrying an oxidizing agent onto the reagent layer. Method of manufacturing ingredients.