JPH04204255A - Composition of printing ink for detecting occult blood in high sensitivity and low sensitivity - Google Patents

Abstract

PURPOSE:To identify negative time and positive time with the presence or absence of coloration by forming this ink of occult-blood detecting composition, adsorbent for adsorbing polar reagent and/or protein material, decay agent, and water absorbing and wetting agent. CONSTITUTION:This ink comprises occult-blood detecting composition, adsorbent for adsorbing polar reagent and/or protein material, decay agent and water absorbing and wetting agent. As the adsorbent for the printing ink composition for detecting occult blood in high sensitivity, the mixed powder of amorphous tricalcium phosphate and crystalline hydroxyapatite is preferable. As the adsorbent for the printing ink composition for detecting occult blood in low sensitivity, the powder of amorphous tricalcium phosphate whose surface is treated with water soluble polymer and/or buffer is preferable. As the decay agent, especially, high water-absorbing resin is used as one component of ink compositions. For example, one or a plurality of the polymers of starch/acrylic-acid graft copolymer, isobutylene/maleic anhydride copolymer and the like can be used. In this way, the negativity, traces and positivity can be identified based on the difference in coloration density.

Description

Detailed Description of the Invention [Industrial Field of Application] The present invention relates to a composition for detecting occult blood and a test specimen, specifically urine,
The present invention relates to a printing ink composition for detecting occult blood for simply and quickly detecting blood in feces and body fluids, and a test body for detecting occult blood formed using this printing ink composition for detection.

[Conventional technology]

Detection of occult blood in body fluids such as urine, vomit, gastric fluid, or fecal excreta is an invaluable tool in the diagnosis of pathological conditions.

Conventionally, to detect blood in body fluids, particularly urine and feces, test specimens coated with occult blood test reagents have been mainly used. This test specimen has the advantage that it is easy to operate and can be used to make a determination in a short time.

The principle of detection in conventional occult blood detection test specimens is that peroxide is decomposed by the peroxidase-like action (an action similar to peroxidase) of hemoglobin in the blood, and the enzyme generated during decomposition reacts with the oxidizable indicator. It is based on the color development of an indicator. However, the peroxidase-like action of hemoglobin in the blood is weak enough to contribute to satisfactory sensitivity and color density in dry chemistry, so in order to solve this problem, quinoline and quinoline derivatives were added to produce peroxidase-like action. A test specimen has been proposed that enhances (in other words, sensitizes) (Japanese Patent Publication No. 41-5680).

However, when the peroxidase-like action is enhanced in this way, satisfactory sensitivity and color density are obtained when using highly sensitive oxidizable indicators such as 0-tolidine and benzidine, but other oxidizable indicators When using a sex indicator, there is a drawback that satisfactory sensitivity and color density cannot be obtained.

Furthermore, the peroxide contained in the specimen for detecting occult blood is decomposed by contact with the indicator and buffer agent contained at the same time, resulting in a decrease in sensitivity and color density. In order to solve this problem and provide long-term storage stability, microencapsulation of organic hydroperoxides using gelatin has been proposed (Japanese Patent Publication No. 10520/1983).

Under these circumstances, the applicant first prints an ink composition in which an oxidizable indicator, an organic peroxide, and a binder are dissolved or dispersed in a solvent onto a support to produce a test object. JP-A-59-33787 for proposing a method), Japanese Patent Application No. 62-75678 for making it possible to manufacture highly sensitive specimens by adding a sensitizer), Capsules of organic peroxide and sensitizer The preservability of the specimen was improved by chemical treatment (Japanese Patent Application No. 62-231779) or addition of an antioxidant (Japanese Patent Application No. 62-222237).

In addition, the use of an inorganic adsorbent as an ink carrier (supporting carrier for the reagent layer) improves the printing suitability of the ink, making it possible to produce test pieces in large quantities. was stabilized, the light resistance of the reagent layer was improved, and quality deterioration during post-processing was eliminated (Patent Application No. 1999).
-182524).

On the other hand, the applicant previously proposed a test piece for detecting occult blood in the OTC urinary direction (KOKAI Publication No. 1-138458) in which a tone layer of a trace color was provided on the outer periphery of the reagent layer on the test piece. In order to manufacture this test piece, a printing ink composition for detecting occult blood (Japanese Patent Application No. 1-182524) containing an inorganic adsorbent (tricalcium phosphate and hydroxyapatite) as an ink carrier was used, and the obtained test The tonal color of the tonal layer (trace; 0.06■ hemoglobin/d!) was determined based on the reagent performance of the piece.
Compared to other test items (sugar, protein, etc.), the color was pale, making it difficult to distinguish between negative, trace, and positive results.

The applicant has also proposed a test piece for detecting fecal occult blood (Japanese Patent Application No. 1-138458) in which a printing ink composition for detecting occult blood is printed directly on a membrane filter.

In this case, a printing ink composition for detecting occult blood containing an inorganic adsorbent (Japanese Patent Application No. 1-182524) was printed directly onto a membrane filter to produce a test piece and evaluated. It was found that the sensitivity was high for use.

[Problem to be solved by the invention]

The object of the present invention is to improve the invention previously made by the applicant in consideration of the prior art, and to provide an occult blood pressure test specimen with excellent storage stability and a low concentration range (trace: 0.06 ■The goal is to develop a product that shows sufficient coloration even when hemoglobin/d is detected and can be identified at a glance compared to a negative test.In other words, even when the test subject (-input) himself tests for urinary occult blood. An object of the present invention is to manufacture an OTC urinary occult blood test strip that can be easily identified by the difference in color density between negative, trace, and positive.

Another objective of the present invention is to develop an OTC device that does not show any color under normal conditions and can distinguish between negative and positive cases by the presence or absence of color, so that it can be easily detected when a subject tests for fecal occult blood. The purpose is to produce fecal occult blood test reagent strips.

[Means to solve the problem]

The present invention solves the above problems, and is characterized by comprising a composition for detecting occult blood, an adsorbent that adsorbs a polar reagent and/or a protein-based substance, a disintegrant, and a water absorption/wetting agent. This is a printing ink composition for high-sensitivity type or low-sensitivity type occult blood detection.

As an adsorbent, a mixed powder of amorphous tricalcium phosphate and crystalline hydroxyapatite is used in a printing ink composition for high-sensitivity type occult blood detection, and a mixed powder of amorphous tricalcium phosphate and crystalline hydroxyapatite is used as an adsorbent, and a printing ink composition for low-sensitivity type occult blood test is used. , amorphous tricalcium phosphate powder surface-treated with a water-soluble polymer and/or a buffering agent is preferred. Furthermore, in a highly sensitive composition, a mixed powder of tricalcium phosphate and crystalline hydroxyapatite treated with a water-soluble polymer and/or a buffer is also preferable as an adsorbent. As a water-soluble polymer,
Preferably gelatin and sodium polyacrylate, or as buffering agents citric acid and sodium citrate, respectively.

In addition, in both pear printing ink compositions, 1. As the disintegrant, a fine powder (400 mesh pass product) of a super absorbent resin, particularly an isobutylene/maleic anhydride copolymer, is preferred. As the water-absorbing/wetting agent, cellulose or cellulose derivatives treated with a water-soluble polymer, particularly low-substituted hydroxypropyl cellulose powder (200 mesh pass product) surface-treated with gelatin, is preferable.

The present invention further provides a high-sensitivity type or low-sensitivity type of occult blood detection, which is characterized in that after adding a binder resin and a solvent to the printing ink composition for occult blood detection, pages are printed on a support. 1 is a method of applying a printing ink composition for printing. in this case,
Ascorbic acid oxidase and/or a chelating agent may further be added to the printing ink composition.

The present invention further provides a test object for detecting occult blood obtained by adding a binder resin and a solvent to the printing ink composition for detecting occult blood, and then directly printing this on a membrane filter. As the membrane filter, a cellulose acetate membrane filter is preferred. Preferably, the membrane filter contains ascorbic acid oxidase and/or a chelating agent and/or a buffer.

As the chelating agent, preferably an iron-coordinated and/or non-iron-coordinated chelating agent is used.

In the present invention, any known composition for detecting occult blood can be used; for example, a composition comprising a microcapsule containing peroxide and quinoline or a derivative thereof, an oxidizable indicator, and a triethanolamine salt is used.

Below, each component of the printing ink composition for detecting occult blood in the present invention and the production of the test specimen for detecting occult blood will be explained in order.

Among the many well-known compounds belonging to the peroxide group, organic hydroperoxides include:
Reacts with the peroxide-active substance in the presence of a peroxide-sensitive indicator to produce a detectable response such as a change in color or a change in the light absorbed by or reflected by the specimen. You can use any of them as long as they are suitable. Examples of those used in the composition for detecting occult blood of the present invention include the following.

Cumene hydroperoxide, t-butyl hydroperoxide, 2,5-dimethylhexane-2,5-dihydroperoxide, diisopropylbenzene hydroperoxide or paramenthane hydroperoxide. These compounds can be used alone or in mixtures. Among these, it is preferable to use cumene hydroperoxide.

The organic hydroperoxide may be contained in an amount of 1 to 30% by weight, preferably 2 to 25% by weight, based on the solid content of the composition for detecting occult blood.

The oxidizable indicator is an indicator that develops color when oxidized by an enzyme, and a wide variety of conventionally known compounds such as benzidines and N-alkylated benzidines can be used. Among these, 0-tolidine, 3.3°, 5.5'
-Tetramethylbenzidine, 2,7-azafluorene and the like are preferred, especially 3.3'.

It is preferred to use 5.5'-tetramethylbenzidine.

This oxidizable indicator is 0.5 to 10% by weight, preferably 0.6 to 6% by weight based on the solid content of the composition for detecting occult blood.
It is desirable that it be present in an amount of

Sensitizer By using a mixture of quinoline or its derivatives and triethanolamine salt as a sensitizer, it promotes color formation of the specimen for detecting occult blood in the presence of blood, improving sensitivity and color density. can do.

As quinoline or its derivative, quinoline, quinine, 6-medoxyquinoline, 6-methylquinoline or 7
-Methylquinoline and the like are used, among which 6-medoxyquinoline is particularly preferred.

As the triethanolamine salt, triethanolamine hydrochloride, triethanolamine phosphate, or triethanolamine sulfate is used, and among these, in particular, triethanolamine lauryl sulfate, which is a sulfate,
Preferably, polyoxyethylene uralyl sulfate triethanolamine is used.

The amount of quinoline or its derivative is 0.5 to 10% by weight, preferably 0.6 to 10% by weight based on the solid content of the composition for detecting occult blood.
Preferably it is present in an amount of 6% by weight.

The triethanolamine salt is desirably present in an amount of 1 to 20% by weight, preferably 2 to 15% by weight, based on the solid content of the composition for detecting occult blood.

Buffer A pH buffer is used to maintain a pH range in which the oxidizable indicator described above undergoes a favorable color change in the composition for detecting occult blood. Any pH buffer may be used as long as it can provide a predetermined pH range (for example, around pH 5) to the composition for detecting occult blood, but specifically, a combination of citric acid and sodium citrate may be used. It is preferable to use it.

Binding material The binding material is required to be one that does not affect the components and 9H etc. in the test liquid, does not affect the reagents, especially the oxidizable indicator, and does not interfere with the color reaction. Ru. Examples of binding materials that have been confirmed to satisfy these conditions include the following, and these materials may be used alone or as a mixture of two or more.

Polyester resin, alkyd resin, polyurethane resin, polystyrene resin, acrylic resin, epoxy resin, vinyl chloride resin, vinyl chloride copolymer resin, polyvinyl butyral resin, polyvinyl alcohol resin, polyvinylpyrrolidone resin or maleic anhydride copolymer resin, etc. synthetic fats: cellulose derivatives such as methylcellulose, ethylcellulose, hydroxyethylcellulose, and carboxymethylcellulose; natural polymers such as starch, polysaccharides, gelatin, casein, or sodium alginate.

The binding material is 0.1 to 2% of the solid content of the composition for detecting occult blood.
It is desirable to use it in a proportion of 0% by weight, preferably 0.5 to 10% by weight.

Disintegrants In the present invention, disintegrants, particularly superabsorbent resins, are used as one component of the ink composition.

This super absorbent resin broadly refers to polymers that absorb and retain large amounts of water several times their own weight, such as starch/acrylic acid graft copolymer system, isobutylene/maleic anhydride copolymer system, polyacrylic acid Salt-based, acrylic acid/
Polymers such as vinyl alcohol copolymers can be used alone or in combination.

There are no particular restrictions on the shape of the resin, but powder, preferably fine powder, particularly 400 mesh bath is advantageous.

As these super absorbent resins, commercially available products can be used as appropriate, and examples thereof include the following, but all types of polymers with high water absorbency can be used. The amount used (compounding amount) can be freely selected in order to adjust the water absorption capacity and adapt the sensitivity, coloring speed, etc. according to the purpose of use of the product, and there are no particular restrictions or 0.0000000000000 yen per long ink composition. About 1 to 30 g is preferable.

Super absorbent resin (200 mesh pass) ■Starch/acrylic acid graft copolymer system (manufactured by Sanyo Kasei) Sunwet IW-300, Sunwet IW-100
0 ■ Isobutylene/maleic anhydride copolymer system (manufactured by Kuraray) KI Gel, 201-F 2 ■ Polyacrylate system (Department of Steel Chemistry) Aqua Keep 4S, Aqua Keep 10SH-NF (manufactured by Arayo Kagaku) Arasorb ( (manufactured by Kao) Boise 5A-20 (manufactured by Sumitomo Chemical) Sumikagel NP-1010 ■Acrylic acid/vinyl alcohol copolymer (manufactured by Sumitomo Chemical) Sumikagel 5P-510, Sumikagel 5P-520 Water absorption/wetting agent Water absorption/wetting agent in the present invention In this method, a water-absorbing polymer such as cellulose, a cellulose derivative, a polysaccharide, a polysaccharide derivative, or a synthetic polymer is used, and a water-absorbing carrier that has not high water retention and wettability is surface-treated (coated) with a water-soluble polymer. Examples of the water-absorbing carrier and water-soluble polymer include the following.

Water-absorbing polymer microcrystalline cellulose, low-substituted hydroxypropyl cellulose, carboxymethyl cellulose-Ca, agarose, viscose, and cross-linked dextrin.

Water-soluble polymers: natural polymers such as gelatin (collagen), casein, keratin, fibroin, soluble starch, gum arabic, sodium alginate, agar, polyvinyl alcohol,
Water-soluble synthetic polymers such as sodium polyacrylate and polyacrylamide, which are insoluble in alcohol solvents such as amyl alcohol and butyl cellosolve acetate, and acetate solvents such as butyl cellosolve acetate.

The water absorbing/wetting agent can be produced, for example, as follows. After dispersing 100 g of water-absorbing carrier in purified water 21, add 0.5 to 20 g of water-soluble polymer, preferably 1 to 1
After adding 0g amount and dispersing it, use a spray dryer (
A water-absorbing/wetting agent having a water-soluble polymer film on the surface of the water-absorbing carrier can be obtained by coating the water-absorbing carrier using a thermal spray dryer.

When using protein-based natural polymers such as gelatin (collagen), casein, keratin, fibroin, and albumin as water-soluble polymers, polyethylene glycol (polymerization degree of 2.000 to
200 preferably 800 to 400), sorbitol, glycerin, polyvinylpyrrolidone (K-15, K-30)
, K-60, K-90, preferably Ni-90) in an amount of 1 to 200% by weight, preferably 10 to 1% by weight, based on the water-soluble polymer.
By adding it in an amount of 0.00% by weight, the performance of the water absorption/wetting agent can be improved.

Surface-treated TPC (granulated powder) Add 3 g of gelatin solubilized in hot water 200 and 1 g of soluble starch to purified water i, sz, and dissolve uniformly. Next, sodium polyacrylate (polymerization degree 2.
700-7,500) 10g, 3g of citric acid, and 12g of trisodium citrate were added and dissolved in order, then 100g of tricalcium phosphate was added, and a homomixer blade for low viscosity (M type homomixer of Tokushu Kikaku Department) was added. used, 12.0
Dispersion treatment is performed for 30 minutes at 0 rpm.

After the dispersion treatment, a coating treatment was performed using a spray dryer (L-8 type spray dryer manufactured by Oyohara Kakoki Co., Ltd.) to form granules (1 to 20 μmφ).
Surface-treated tricalcium phosphate with high fluidity is obtained.

Adsorbent The adsorbent used in the present invention can be any inorganic or bred material as long as it has the property of adsorbing a coloring reagent and/or a protein substance (enzyme, antigen, hubten, antibody, protein, etc.). It can be of any type, such as the following: It is preferably used in powder form, particularly fine powder form. These adsorbents may be used alone or as a mixture of two or more, or several types of the same type may be used in combination. The amount used is not particularly limited, but is preferably about 10 to 40 g per 100 g of the ink composition.

Adsorbents (microcarriers) that retain proteins and polar drugs;
0.1 to 100 μmφ): ■Inorganic porous material Porous glass, silica gel, alumina, activated clay, quartz sand, magnesium oxide, magnesium carbonate, bentonite, kaolin, talc, hydroxyapatite, tricalcium phosphate, titanium oxide, activated carbon.

■Breeding porous materials Nylon 6 powder, nylon 6.6 powder, tetrafluoroethylene powder, polyvinylidene difluoride powder, trifluorochloride ethylene powder, and plasma-treated products of the above powders.

■Supports with polar groups (anionic, cationic or zwitterionic) Anion exchange resins, cation exchange resins, macroporous ion exchange resins, hybrid ion exchange resins, ion exchange fibers, chelate resins.

Example: DEAE - ground cellulose, ion exchange resin for high performance liquid chromatography.

■Polymer flocculants Nonionic polymer flocculants, polycationic polymer flocculants, polyanionic polymer flocculants, amphoteric polymer flocculants.

Examples Chitosan, polyaminoalkyl methacrylate (and copolymers with acrylamide).

■Examples of polymers in which adsorbed substances are immobilized by chemical treatment Antibody-immobilized polymers, lectin-immobilized polymers, anti-, antigen- or antigen-hapten-immobilized polymers (Polymers include cellulose, agarose, etc.) , dextran, etc.).

■ Polymer carrier synthesis for separation (chromatography) Polymer gel filtration applicable carrier, polysaccharide gel filtration applicable carrier.

Example: Support for high performance liquid chromatography.

In addition to the above five types of adsorbents, the following are also exemplified.

■ Microplastics such as latex formed by copolymerization are treated with water-soluble salts (phosphates, calcium salts, barium salts, etc.).

■Water-insoluble oxides (titanate, alumina, etc.) are treated with ion (anion, cation, cation) exchanged cellulose fine powder.

In the printing ink composition of the present invention, it is important that an adsorbent is used to adsorb drugs, etc. in a solvent to the adsorbent.

In the present invention, since drugs and the like are immobilized in a solution, they only need to have a polar group, and the adsorbent advantageously utilizes this to adsorb reagents and protein substances. Hereinafter, the present invention will be described using an insoluble porous finely powdered salt, particularly a calcium phosphate salt, as a representative example of the adsorbent, and the same applies to other types of adsorbents.

The solvent is preferably one that can uniformly and stably dissolve or disperse each of the above components, especially the binder. Examples of solvents that meet these conditions include: ■ aromatic hydrocarbons such as benzene and toluene, ■ aliphatic ketones such as methyl ethyl ketone and cyclohexanone, ■ esters such as ethyl acetate, and ■ alcohols such as n-butanol. Non-aqueous solvents such as water,
One type or a mixture of several types from these ① to ② can be used.

However, it is preferable to use a non-aqueous solvent because the drying step after coating the composition on the support can be carried out at a low temperature and in a short time.

Furthermore, the use of a non-aqueous solvent contributes to maintaining a highly stable reagent composition state and also has the effect of improving the thermal stability of the enzyme.

Other Components Depending on the case, a wetting agent may be incorporated into the composition for detecting occult blood in addition to the above-mentioned components.

Wetting agents include nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, and polyethylene glycols. Wetting agents can help disperse each reagent, promote the formation of a uniform test area, and improve water wettability. Preferably, the wetting agent is present in an amount of 0.5 to 5% by weight based on the solids content of the composition for detecting occult blood.

Further, in order to make the color tone of the indicator more visible, a background dye such as oil yellow may be added.

Microcapsule coating material The microcapsule coating material that can be used in the present invention is as follows:
The components constituting the composition for detecting occult blood are a first group consisting of an organic hydroperoxide, a sensitizer, and a part of a binder, an oxidizable indicator, a pH buffer, a water-absorbing powder, and a part of a binder. If the first group is divided into a second group consisting of
An emulsifier that forms a good emulsion when mixed with any of the components belonging to the above group or with any of the components belonging to the second group, and which quickly disintegrates when it comes into contact with water in the test liquid and eliminates the encapsulation. It is desirable that the organic hydroperoxide or oxidizable indicator, etc., which has been present in the reaction system, be released to participate in the color reaction.

Examples of microcapsule coating materials that have been confirmed to satisfy these conditions include the following.

Water-soluble synthetic resins such as polyvinyl alcohol resin, polyvinylpyrrolidone resin, polyacrylamide resin or polyvinyl acetate resin; Water-soluble cellulose derivatives such as methylcellulose, ethylcellulose, hydroxycellulose or carboxymethylcellulose: starch, polysaccharides, gelatin, casein, gum arabic or natural polymers such as sodium alginate.

Among these, gum arabic, carboxymethyl cellulose and polyvinyl alcohol resin are particularly preferred.

The microcapsule coating material is preferably present in an amount of 2 to 70% by weight, particularly 4 to 50% by weight, based on the solid content of the composition for detecting occult blood.

Manufacture of Test Body The composition for detecting occult blood as described above is applied as an ink composition onto a support, and then dried to form an area for detecting occult blood, and a test body is obtained.

As a coating method, a printing method, a coating method (for example, a roll coating method, a spray coating method, a dip coating method, a solid coating method), etc. are used.

In the present invention, the coating amount of the composition for detecting occult blood is preferably relatively large and constant, so the composition for detecting occult blood is coated on the support by silk screen printing, plate printing, gravure printing, etc. It is preferable to apply it to

The amount of the composition for detecting occult blood to be coated on the support depends on the type of the composition, but is generally preferably from 2 to 150 g/rr'lt (dry).

Support (1) The support is preferably one that does not react with the composition for detecting occult blood and does not inhibit the coloring of the indicator.

Specifically, it is paper, synthetic paper, nonwoven fabric, synthetic resin film, or a composite thereof, such as a laminate such as a combination of paper and synthetic resin film.

The above support that fosters ultrafiltration action (pore size 0, O1
~10 μmφ) can also be used similarly.

A test object having an occult blood detection area formed on such a support may be formed into a stick shape, a roll shape, a tape shape, or the like. In addition, the support itself can be in a shape that allows the test liquid to be collected, such as a pot shape, a test tube shape, a dish shape, etc.
It may be formed into a tray shape or a dropper shape.

Support (2) The membrane filter used in the present invention has a pore size of 0.01 to 10 μmφ, is resistant to shrinkage in hot air at 100°C (is stable to alcohol solvents, and has sufficient mechanical strength as a printing base material). If it has, cellulose type, vinyl type, aromatic type and all other types can be used.

(1) Examples of hydrophilic membrane filters with low adsorption of proteins and drugs: Nylon 6 membrane filter, Nylon 6.6
Membrane filter, regenerated cellulose membrane filter, polyether sulfone membrane filter,
Cellulose acetate membrane filter, etc.

According to the present invention, super absorbent resin fine powder, for example, KI
By using Gel 201 in combination with -F and/or Sumikagel 5P-510, the color uniformity can be improved while maintaining high reaction sensitivity in the test area of the test specimen for detecting occult blood, and the light resistance can be significantly improved. In addition, a printing ink composition for detecting occult blood that has good printability and high productivity can be prepared.

Tricalcium phosphate (TPC) powder is a highly cohesive amorphous fine powder that is insoluble in water but hydrophilic, and is a bipolar physical adsorbent with phosphate and calcium groups. . Tricalcium phosphate is not only used as an excellent ink carrier that exhibits good printing suitability in mixed solvents of alcohols such as amyl alcohol and screen printing solvents such as butyl cellosolve acetate, but also as an oxidizable material with polar groups. To improve the dispersibility of the indicator in the ink and stabilize the indicator by adsorbing the color indicator in a solvent, and to significantly improve the light resistance of a printing ink composition and a specimen obtained from the same without impairing coloring performance. The stability (lightfastness) of an oxidizable color indicator depends on the weight ratio of this indicator to TPC in the printing ink composition, with a specific oxidizable color indicator of 3.3'.

Approximately 10 in the case of 5.5゛-tetramethylbenzidine
Double weight TPC is preferred. As the weight ratio of TPC increases, the stability of the indicator improves or the coloring sensitivity decreases. However, unlike the conventional adjustment of color sensitivity by adding an ordinary antioxidant or substituting a low-sensitivity indicator, the composition of the present invention using TPC has achromaticity in the insensitive region and color sensitivity in the sensitive region. Excellent coloring performance. That is, by adjusting the weight ratio of the indicator and TPC, the stability of the indicator can be adjusted, and the color sensitivity can also be adjusted. Furthermore, as mentioned earlier, due to the physical properties of TPC, which is a non-crystalline powder with strong cohesiveness, when used alone as an ink carrier, it causes wetting and penetration of the test area (reagent layer) of the manufactured specimen. Poor quality and cracking may occur. As a solution to this problem, three methods described below have been established.

Method 1: Add powder with low cohesiveness.

Example MT-100S (15 nm φ fine particles of titanium oxide treated with a fatty acid mainly consisting of lauric acid and an AI compound, manufactured by Teika ■).

Method 2: TPO is surface treated with a water-soluble polymer and/or buffer as described above and granulated.

Method 3゜ Combined use with sintered hydroxyapatite powder.

To prepare a printing ink composition for highly sensitive occult blood testing,
It is preferable to use sintered hydroxyapatite powder as an ink carrier (Method 3), and to prepare a printing ink composition for low-sensitivity occult blood testing, surface-treated and granulated TPO (Method 2) is preferable. It is suitable to use it alone as an ink carrier.

Hydroxyapatite (RAP) powder is a calcium phosphate salt with a hydroxyl group and has crystallinity, and has good wettability with an aqueous solution. When used in combination with TPC as an ink carrier, it improves the wettability of the test area of a specimen for detecting occult blood. Maintain high reaction sensitivity and color density. As HAP powder,
It is preferable to use a porous sintered product.

Due to its strong and rapid swelling performance, superabsorbent resin
It is very effective as a disintegrant for test specimens using highly cohesive, non-swellable ink carriers. The particle size of the super absorbent resin is 400 mm, considering both printability and disintegration performance.
A mesh pass product with a particle diameter of more than 10 μmφ is suitable. In addition, super absorbent resin is super hydrophilic, so it does not absorb solvents, which prevents ink from becoming bulky, and has good printability.
Does not interfere with the function of the adsorbent, which takes in reagents etc. dissolved or dispersed in the solvent. However, although the superabsorbent resin has high water absorption performance, it is difficult to release the sample liquid that has been absorbed, and it does not have the effect as a wetting agent that increases the water retention of the test area.

Low-substituted hydroxypropylcellulose (GT-L-RPC) coated with gelatin as a water-absorbing and wetting agent does not absorb solvent components in the ink, and does not reduce the light resistance of the ink and the specimen. In sample liquid, it swells with a water absorption capacity of several times its own volume, but it freely releases the absorbed sample liquid, producing a wetting effect.

In order to increase the water absorption and wettability of the test area of the specimen and to increase the color sensitivity, it is preferable to use a disintegrant and a water absorption/wetting agent in combination.

〔Example〕

EXAMPLES Hereinafter, the preparation of the printing ink composition for detecting occult blood of the present invention and the manufacture of test specimens using these printing ink compositions will be explained in detail with reference to Examples.

Example 1 Printing ink composition for highly sensitive occult blood detection (margins below this page) (for OCT urinary occult blood test specimen) A. Composition table of printing ink composition for occult blood ■, TM
B 1.33■, mixed solution 26.80 [BCAc
8.001 [20% PEG2,000
(in Amyl OH) 9.931 [20% Emal TD (in Acnyl OH) 8.87] ■
, 10% MBBC (in BCAc)
2.22■, TPC12,59 ■, sugar buffer 9,49■,
KI gel 201-F2 0.8
9■, (3%GT)-L-1 (PC4,43■, HA
P (200 mesh pass product) 11.52■
, binder resin 15.78[Am
yl O) (9,15] [BCAc 3.89] [PVP
2゜321[BX-10,4
15] [Phase], MC15,00 1 cumene hydroberoquinde
3.38][6-Medquinquinoline
1.13) Total 100.05 Abbreviations in the table have the following meanings.

TMB ・3.3',5.5'-Tetramethylbenzidine PEG: Polyethylene glycol MBBC: 2,2°-methylenebis-(6-tert-
butyl-p-cresol) TPC Tricalcium diphosphate GT: Gelatin L-HPC: Low substituted anti-type hydroxypropyl cellulose HAP: Hydroxyavatite MC: Microcapsule B, Preparation method of ink composition Pre-prepared solid emeru TD Emeru TD (lauryl sulfate triethanolamine salt, manufactured by Kao), available on the market for preparation, is a 40% aqueous solution and cannot be used directly in ink. Therefore, the following treatment is performed to obtain a solid material.

Emeru TD 10 on the market in a stainless steel tray
Add 50 parts by weight of 95% ethanol to 0 parts by weight, and use an explosion-proof oven to heat the mixture at 60-80°C for about 2 hours to dehydrate it. (stir). This process is repeated three times, and after cooling, a solid Emmaru TD is obtained.

Preparation of HAP and L-HPC for 200 mesh baths (approximately 70 μm or less) HAP (hydroxyapatite for column chromatography; Wako Pharmaceutical Co., Ltd.) and L-HPC (low-substituted hydroxypropyl cellulose Ll (-31; Shin-Etsu Chemical Co., Ltd.) ) is a 200 mesh residual component of 10-20%・(W
/W), so it cannot be used directly in ink. Therefore, sieving is performed to obtain a 200-mesh bath product (improved printability).

The powders of TPC, HAP and L-HPC used for the preparation of dry powders are:
Drying is performed at 100°C for 3 hours under reduced pressure using a vacuum oven.

Preparation of MBBC (Antioxidant) Solution 10% (w/total w++) MBBC (in BCAc) is solubilized by sonication.

Preparation of mixed solution 20% (w/total w) PEG2.000 (Amyl O
11.20 parts by weight (in Amyl OH) and 10.00 parts by weight of 20% (w/total w) Emal TD (in Amyl OH) and BC
9.00 parts by weight of Ac is placed in a plastic container, sealed and heated to obtain a clear solution.

MC treatment of organic peroxide (cumene hydroperoxide) Cumene hydroperoxide and 6-medoxyquinoline, which is a sensitizer, are converted into MC using a spray dryer using gum arabic as a coating agent (Japanese Patent Application No. 62-23177
No. 9).

This preparation 1) Add ■ to a round Tupperware (880 ml), heat it (50-60°C) to make a clear solution, and then
After adding and performing dispersion treatment using an ultrasonic cleaner,
A heating treatment is carried out again (0.5-1 hours) to obtain a similarly clear solution.

2) After adding ■ to the above solution and stirring and mixing, the solution is returned to room temperature and then degassed using a vacuum pump (5 minutes), followed by a heating process after sealing (1 hour).

3) After returning the above solution to room temperature, mix with stirring using an agitator, and then add (1), (2), (2), (2), (2) and [phase] in order to prepare an ink composition.

An outline of the process for preparing the above ink composition is shown below.

(Margins below this page) After printing the test reagent part in the form of stripes with a width of 5 mm on a white polystyrene sheet with a thickness of 250 μm using the ink composition I for detecting occult blood prepared according to the above preparation method using the screen printing method. , and dried for 20 minutes at a temperature of 60°C. The number of lines of the plate used for printing was 120 meshes, and the total thickness of the resist and screen gauze was 700 μm. After drying, it was cut into sticks of predetermined dimensions to obtain a detection test piece I.

C1 color performance test The following four types of liquids were prepared as test liquids.

■ Normal urine (-) ■ Urine containing 0.06■/dl of human hemoglobin (manufactured by Sigma) (±): Trace value■ Urine containing human hemoglobin (manufactured by Sigma) of 0.15■/di (Kano) ■ Urine containing 0.75■/dl of human hemoglobin (manufactured by Sigma) (±) The detection test piece obtained in B above was immersed in the test liquids of ■ to ■, and immediately removed after immersion, it was left to stand for 1 minute. The color development of the detection reagent portion after the test was placed was observed.

No negative color development was observed in the detection reagent part. In addition, the color of the detection reagent part is bright and clear, and the color density increases stepwise as the human hemoglobin concentration in the test fluid increases, and within the above range, occult blood in the test fluid is detected. The concentration could be clearly determined.

No change in color tone was observed even after the colored detection test piece was allowed to stand at room temperature for 5 minutes.

In addition, the obtained detection test piece was sealed in a glass container,
After storage at 40°C for 12 months, a test was performed in the same manner as above, and the color of the test reagent was uniform, clear, and stable as before storage, and the human hemoglobin concentration in the test solution was It was possible to clearly distinguish them as before storage.

In addition, in addition to the test subjects described in ■ to ■ above, normal urine and normal urine contain 20 cells/μl and 50 cells/μl of red blood cells, respectively.
1 and 250 particles/μl were prepared as a test solution and colored in the same manner.
Clear and stable spots are formed, and the number of spots gradually increases as the concentration of red blood cells in the test fluid increases, and the concentration of red blood cells in the test fluid can be clearly determined within the above range. there were. No change in color tone was observed even after the colored detection test piece was allowed to stand at room temperature for 5 minutes.

In addition, the obtained detection test piece was sealed in a glass container,
After storage at 40°C for 12 months, a test was performed in the same manner as above, and the coloration of the test area was as clear and stable as before storage, indicating that the concentration of red blood cells in the test solution had been preserved. As before, it was possible to distinguish clearly.

D. Ink composition for occult blood color tone I The above detection test piece was immersed in 0.06 x 7 dl of urine containing human hemoglobin (+: trace), and the test reagent part was taken out immediately after immersion and left to stand for 1 minute. An ink composition for toning was prepared with a toning density of .

(Left space on this page) Composition table of ink composition I for occult blood tone Patch size
Per 100 parts by weight of ink (parts by weight) Sumikagel 5P-52030,00 Sericol Arashi 13 56.00
Butyl cellosolve (BC) 12.5
60.67% Kayset Blue 714 (
Cyan color 2 BC') 1.08 The above color tone ink compositions were 3% each compared to the color tone ink composition of the detection test piece S prepared by the method described in Japanese Patent Application No. 1-138458. Double amounts of dye solution (cyan and yellow) were required.

Example 2 Printing ink composition for low-sensitivity type occult blood detection ■ (for OTC fecal occult blood test specimen) (Margins below this page) A. Composition table of printing ink composition for occult blood Patch size Per 100 parts by weight of ink ( (parts by weight) ■, TMB 1.
29 ■, mixed solution 27.68
■, 10% MBBC (in CHN)
2.77■8 Surface treated TPC (granulated form)
32.29■, Kl gel 201-F2
0.92■, (3% GT)-L-HPC
4,61■, Resin for binding material (composition is the same as I) 1
5.68 Total 100.00 C) IN: Cyclohe + Sunon Ink composition for detecting occult blood was prepared in the same manner as in Example 1, and using this in the same manner as in Example 1 B. A detection test piece (■) was obtained by screen printing. Next, using this test piece, a coloring performance test was conducted in the same manner as C in Example 1.

The results of the coloring performance test of the test piece I according to Example 1 and the test piece ■ according to Example 2 are shown in the table below.

For comparison, the results of test piece S according to Japanese Patent Application No. 1-13845, which was similarly tested, are also shown in the table. The reaction time is 1 minute in each case.

Regarding both color development sensitivity and color development speed, the performance of test piece ■ was lower than that of test piece S in the low concentration range (hHG O,06 [Dg/d/or less)], but in the high concentration range (hHG 0.75 [Dg/d/or less)]
mg/dl or higher) showed equivalent performance. When the storage test was conducted on the test piece (■) in the same manner as the test specimen, the same excellent results were obtained.

By the way, TMB is a highly sensitive, oxidizable color indicator that is extremely unstable to light, and remains stable for less than 1 hour in the air, even indoors under fluorescent light, and turns green (oxidized). . However, an improvement in light resistance was observed in the test pieces produced in combination with TPC. Test pieces S and ■ use a combination of amorphous TPC and crystalline calcium hydroxide phosphate (hydroxyapatite: HAP) as ink carriers, and these test pieces have high stability under normal storage conditions; It is stable for several hours if left in the air under indoor fluorescent lighting, or changes in color (greening) if left for a day. However, the test piece (2) using granulated TPC as an ink carrier showed some discoloration (browning) but did not turn green even when left in the air under an indoor fluorescent lamp.

These results suggest that the stabilization of TMB is more important than the protective effects such as the hiding effect of amorphous TPC or the incorporation of HAP into the porous interior, and rather the binding of the phosphate group and TMB to stabilize this indicator. It is recognized that it brings about

As described above, ink composition (1) has very high storage stability, is extremely unlikely to cause negative color development, and exhibits uniform and good color development for high concentrations of hemoglobin (0.75 mg/d! or more).

Example 3 Test specimen for detecting fecal occult blood (margins below this page) A. Composition table of spreading layer printing ink composition (parts by weight) ■9 Surface-treated TPC (granules) 40.
0■, Kr gel 201-F2
' 1.5■4 Span 20
1.5 ■, CHN
9.0■, Amyl OH21.0 ■6 Resin for binder 27.0 Total 100.0 CHN: Ncrohexanone coated type membrane filter (Toyo Roshi Department,
Tetron 120 mesh: Film thickness 2
(0 μm plate used) and dried at 60°C for 20 minutes. Next, a square test reagent part with a side of 7 mm was printed directly on the back side using printing ink composition (2) for low-sensitivity type occult blood detection by screen printing, and dried at 60°C for 20 minutes. After drying, it was cut into a square of 15 m on each side with the reagent part in the center, and this was stacked and fixed on a mount (filter paper) with square holes of loam width so that the mount and the reagent part faced each other. A detection test piece was obtained. This detection test piece is shown in FIG. 1 as a cross-sectional view. At this time, a wetness confirmation layer may be provided on the outer layer of the test reagent part using a binder resin containing cobalt chloride as an indicator of permeation of the sample liquid.

Example 4 Test body A for detecting fecal occult blood, support (water-soluble paper) Both sides of a water-soluble paper (manufactured by Mishima Paper; Disolpo CD-2, neutral, thickness 160 μm) are coated with the following resin B.

B. Coating resin GANTREZ-AN-169 (manufactured by G, A, G; methyl vinyl ether/maleic anhydride copolymer) 1
5 parts by weight of n-pentanol is heated in 85 parts by weight of n-pentanol to convert maleic anhydride groups into carboxyl groups and pentanol ester groups (half ester), and this resin mixture is added with BC.
5 parts by weight of Ac114 and 11 parts by weight of fine TiO□ (manufactured by Teika, MT-100s) surface-treated with stearic acid are added.

The printing ink composition for a spreading layer described in Example 3 is printed solidly on one side of support A by screen printing (plate T1
20B, film thickness 20 μm). A reagent ink layer (low sensitivity type) and a wetness indicator layer are printed on this developing layer as shown in FIG. FIG. 2A is a top view of the obtained occult blood detection utility body, and FIG. 2B is a sectional view thereof.

[Effects of the Invention] The present invention is an improvement of the printing ink composition for detecting occult blood described in Japanese Patent Application No. 1-182524. Compositions for low-sensitivity type and test specimens using them are provided.

In the printing ink composition for highly sensitive occult blood testing of the present invention, various components are uniformly dispersed by heat treatment, physical treatment such as ultrasonic waves, and adjustment of the solvent ratio and amount. The interaction between the sensitizer and the adsorption carrier is further improved. At the same time, according to the present invention, stabilization and uniform dispersion of the color reagent, and increase in the amount of the color reagent and sensitizer (i.e., increase in sensitivity) under conditions where the printing ink composition is stable. It is possible to achieve both. Furthermore, according to the present invention, stable uniform dispersion of the binder resin is improved by using a mixed solvent in a specific ratio under specific addition conditions of the superabsorbent resin as a disintegrant, and light resistance is improved. The printability, light fastness and storage stability of printing ink compositions and specimens can be improved by using a water-absorbing/wetting agent that does not reduce the properties (i.e. does not swell in the solvent containing the solubilized coloring reagent). Stability in negative conditions (no color development) and color sensitivity in low concentration regions (trace) are enhanced without loss. In addition, by changing the weight ratio of 3,3',5.5'-tetramethylbenzidine, an oxidizable indicator, and tricalcium phosphate, an inorganic adsorbent, as an ink carrier, the color sensitivity can be finely adjusted. It can be adjusted.

In the printing ink composition for low-sensitivity type occult blood detection of the present invention, the same excellent effects as those obtained in the above-mentioned high-sensitivity type can be obtained mainly by selecting and/or modifying the adsorbent as an ink carrier. or achieved. More specifically, the color reaction of the specimen to be examined is suppressed in a region of low occult blood concentration, or uniform and good coloration is obtained at occult blood concentrations exceeding a threshold value. This is not achieved by conventional means such as adding antioxidants or replacing high-sensitivity color reagents with low-sensitivity color reagents, but by using the inorganic adsorbent (TPC) of the present invention.
) physically adsorbs the highly sensitive coloring reagent and stabilizes the coloring reagent, thereby increasing the oxidation range of the coloring reagent,
It is possible to effectively suppress color reactions in low concentration areas of occult blood. Furthermore, the uniformity of color development can also be improved by surface treating TPC with a water-soluble polymer and a buffer.

[Brief explanation of the drawing]

FIG. 1 shows one embodiment of the test object for detecting occult blood of the present invention, and is a sectional view of the test object manufactured according to Example 3. FIG. 2 shows another embodiment of the test object of the present invention (Example 4)
, A is a top view thereof, and B is a sectional view thereof. Figure 2A Figure 2B Mouth (green area): Occult blood detection reagent layer

Claims (1)

[Claims] 1. A highly sensitive type or Printing ink composition for low-sensitivity occult blood detection. 2. The composition for occult blood testing comprises a composition for occult blood testing, an adsorbent, a disintegrant, and a water-absorbing/wetting agent, and the adsorbent is a mixed powder of amorphous tricalcium phosphate and crystalline hydroxyapatite. A printing ink composition for highly sensitive occult blood detection. 3. Composition for occult blood testing, consisting of an adsorbent, a disintegrant, and a water absorption/wetting agent, where the adsorbent is amorphous tricalcium phosphate powder whose surface has been treated with a water-soluble polymer and/or a buffering agent. The printing ink composition for low-sensitivity type occult blood detection according to claim 1. 4. Composition for occult blood testing, consisting of an adsorbent, a disintegrating agent, and a water absorption/wetting agent, where the adsorbent is composed of amorphous tricalcium phosphate whose surface has been treated with a water-soluble polymer and/or a buffer, and a crystalline tricalcium phosphate. The printing ink composition for highly sensitive occult blood detection according to claim 1, which is a mixed powder of hydroxyapatite. 5. Composition for occult blood testing, consisting of an adsorbent, a disintegrant, and a water absorbing/wetting agent, and the adsorbent is coated with a water-soluble polymer of soluble starch, gelatin, and sodium polyacrylate, and a buffering agent of citric acid and sodium citrate. The printing ink composition for high-sensitivity type or low-sensitivity type occult blood detection according to claim 3 or 4, which is treated amorphous tricalcium phosphate. 6. Consists of a composition for occult blood testing, an adsorbent, a disintegrant, and a water absorbing/wetting agent, and the disintegrant is a fine powder of a super absorbent resin (400
The printing ink composition for high-sensitivity type or low-sensitivity type occult blood detection according to claim 1, which is a mesh pass product). 7. Composition for occult blood testing, consisting of an adsorbent, a disintegrant, and a water-absorbing/wetting agent, where the water-absorbing/wetting agent is a powder of cellulose or cellulose derivative surface-treated with a water-soluble polymer (20
The printing ink composition for high-sensitivity type or low-sensitivity type occult blood detection according to claim 1, which is a 0 mesh pass product). 8. Composition for occult blood testing, consisting of an adsorbent, a disintegrant, and a water-absorbing/wetting agent, the disintegrant being a fine powder of isobutylene/maleic anhydride copolymer (400 mesh pass product), and the water-absorbing/wetting agent The printing ink composition for high-sensitivity type or low-sensitivity type occult blood detection according to claim 6 or 7, wherein is a low-sensitivity type hydroxypropylcellulose powder (200 mesh pass product) surface-treated with gelatin. 9. High sensitivity, characterized in that after adding a binder resin and a solvent to the printing ink composition for detecting occult blood according to any one of claims 1 to 8, this is directly printed on a support. A method of applying a printing ink composition for type or low sensitivity type occult blood detection. 10. A specimen for detecting occult blood obtained by adding a binder resin and a solvent to the printing ink composition for detecting occult blood according to any one of claims 1 to 8, and then directly printing the mixture on a membrane filter. . 11. After adding a binder resin and a solvent to the ink composition for detecting occult blood according to any one of claims 1 to 8, this is mixed with a metal-free chelating agent and/or an iron-coordinated chelating agent. and/or a test object for detecting occult blood obtained by printing directly onto a membrane filter containing a buffering agent. 12. After adding a binder resin and a solvent to the printing ink composition for detecting occult blood according to claim 3, this is mixed with an iron chelating agent (
A test object for detecting occult blood obtained by directly printing on a cellulose acetate membrane filter containing HEDTA or EDTA) and/or a buffer. 13. After adding a binder resin and a solvent to the printing ink composition for detecting occult blood according to claim 3, this is mixed with a chelating agent (HEDTA-Fe^+^+^+ or E
DTA-Fe^+^+^+, HEDTA-Cu^+^+
or EDTA-Cu^+^+) and/or a buffer for detecting occult blood by directly printing on a cellulose acetate membrane filter. 14. After adding a binder resin and a solvent to the printing ink composition for detecting occult blood according to claim 3, this is mixed with a chelating agent (HEDTA-Fe^+^+^+ or E
DTA-Fe^+^+^+, HEDTA-Cu^+^+
or EDTA-Cu^+^+), a chelating agent (HEDTA or EDTA) in which no metal is coordinated, and/or a buffer agent. 15. After adding a binder resin and a solvent to the printing ink composition for detecting occult blood according to any one of claims 1 to 8, this is mixed with ascorbic acid oxidase and a metal-free chelating agent and/or A specimen for detecting occult blood obtained by printing directly onto a membrane filter containing an iron-coordinated chelating agent and/or a buffering agent. 16. After adding a binder resin and a solvent to the printing ink composition for detecting occult blood according to claim 3, this is mixed with ascorbic acid oxidase and an iron chelating agent (HE) in which no metal is coordinated.
DTA and/or EDTA) and/or iron- or copper-coordinated chelating agents (HEDTA-Fe^+^+^+ and/
Or EDTA-Fe^+^+^+, HEDTA-Cu^
A specimen for detecting occult blood obtained by directly printing on a cellulose acetate membrane filter containing EDTA-Cu^+^+) and/or a buffer. 17. After adding ascorbic acid oxidase, an iron chelating agent, a binder resin, and a solvent to the printing ink composition for detecting occult blood according to any one of claims 1 to 8, this is directly printed on a support. A method for applying a printing ink composition for detecting high-sensitivity or low-sensitivity occult blood, characterized by: 18. The printing ink composition for detecting occult blood according to any one of claims 1 to 8 contains iron (Fe^+^+^+) or copper (Cu
A high-sensitivity type characterized by adding an iron chelating agent coordinated with ＾+＾+), an uncoordinated iron chelating agent, a binder resin and a solvent, and then printing these directly onto the support. Or a method of applying a printing ink composition for detecting low-sensitivity occult blood.