JPH01147366A - Composition for detecting occult blood and inspecting body using said composition - Google Patents

Abstract

PURPOSE:To improve detection sensitivity and coloration sensitivity and to enable semiquantitative determination of hemoglobin and red cells in a low concn. region by adding quinoline and ethanol amine salt to a compsn. for detecting occult blood. CONSTITUTION:The compsn. for detecting occult blood is prepd. by microencapsulating a peroxide such as cumene hydroperoxide and sensitizer such as 6-methoxyquinoline by using gum arabic, then adding an indicator to be oxidized such as 3, 3', 5, 5'-tetramethyl benzine, pH buffer, binder, water absorbent powder, solvent and swelling agent thereto. This compsn. is finely dispersed by a homomixer and is printed on a white polystyrene sheet by a screen printing method and is dried. The inspecting piece prepd. in such a manner is immersed in a liquid to be inspected and is taken out immediately after inspection. The coloration of the inspection reagent part after resting for 1min is then observed.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to a composition for detecting occult blood and a test specimen, in particular,
The present invention relates to a detection composition for simply and quickly detecting blood in urine, feces, and body fluids, and a test body formed using the detection composition.

BACKGROUND OF THE INVENTION The detection of occult blood in body fluids such as urine, gastrointestinal fluids, or fecal excreta is an invaluable tool in the diagnosis of many pathological conditions.

Conventionally, to detect white fluid in body fluids, especially urine, 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 make a determination in a short time.

However, in conventional occult blood detection test specimens, peroxide is decomposed by the peroxidase-like action (an action similar to peroxidase) of hemoglobin in the blood, and the oxygen generated during decomposition reacts with the oxidizable indicator. It is based on the principle of coloring an indicator, and the peroxidase-like action of hemoglobin in the blood is weak to contribute to satisfactory sensitivity and color density in dry chemistry. A test piece containing a derivative to enhance the peroxidase-like action (in other words, to sensitize it) has been proposed (Japanese Patent Publication No. 5680/1983).

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

Furthermore, the peroxide contained in the occult blood test strip is also decomposed by contact with the indicator and M buffer 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 (by line hydroperoxides) using gelatin has also been proposed (Japanese Patent Publication No. 10520/1983). .

Under such circumstances, the applicant first dissolves or disperses the oxidizable indicator, the row peroxide, and the binder,
proposes a method of manufacturing a test object by printing an ink composition on a support (Japanese Patent Application Laid-Open No. 59-33787)
.

Although this method has a simplified manufacturing process and can efficiently manufacture test pieces, it is not necessarily sufficient in terms of storage stability and sensitivity.

K Problems to be Solved by the Invention] The present invention improves the invention previously made by the applicant in consideration of the prior art, and provides a composition for occult blood testing and a test specimen;
1) It has excellent storage stability, 2) It can be dried at low temperatures and does not cause deterioration of the indicator, and 2) The coating method, especially printing. This was done with the aim of obtaining something that satisfies all the requirements that an inspection area can be formed by the method.

Means for Solving Problem K) In order to solve the above problems, the present invention uses a specific sensitizer to achieve high sensitivity and color density, and is easily manufactured by means such as printing. A composition for detecting occult blood and a specimen were obtained.

The present invention is a composition for detecting occult blood that can give a detectable response in the presence of a peroxidizing active substance, comprising a peroxide, an oxidizable indicator, and quinoline or m 1 thereof.
The present invention relates to a composition for detecting occult blood characterized by containing a body and a triethanolamine salt, and a test body for detecting occult blood obtained by coating the composition on a support.

More specifically, the composition for detecting occult blood according to the present invention comprises a peroxide, an oxidizable indicator capable of making a detectable response in the presence of a peroxide and a peroxidizing active substance, and the above-mentioned quinoline or a derivative thereof. A composition for detecting occult blood in which a sensitizer consisting of a triethanolamine salt, a pH buffer, a binder, and a water-absorbing powder are dissolved or dispersed in a non-aqueous solvent, and if necessary, the constituent components into a first group containing a peroxide and a second group containing an oxidizable indicator, and either the first group or the second group is microencapsulated with a water-soluble microcapsule coating material. You can also.

K effect] According to the present invention, by combining quinoline or a quinoline derivative, which is a known sensitizer, with triethanolamine salt, which is a sensitizing agent, the sensitivity of the reaction of the composition for detecting occult blood can be significantly increased. It can be made higher.

EXAMPLES Below, each component of the 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.

Peroxides Among the many well-known compounds belonging to the peroxide group, there are:
A peroxide-active substance that reacts with a peroxide-active substance in the presence of a peroxide-sensitive indicator to produce a detectable response, such as a color change or a change in the light absorbed or reflected by the specimen. You can use any of them as long as you can. Examples of substances 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, para-menthane hydroperoxide, etc. alone, or a mixture thereof.

Among these, it is preferable to use cumene hydroperoxide.

B1 hydroperoxide can 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.

Oxidizable Indicator 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, etc. are preferred, especially 3.3', 5.5'
-Tetramethylbenzidine is preferably used.

This oxidizable indicator is desirably present in an amount of 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.

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 24 forms, quinoline, quinine, 6-medoxyquinoline, 6-methylquinoline, or 7-methylquinoline are used, among which 6-
Particularly preferred is medoxyquinoline.

As the triethanolamine salt, triethanolamine sag, triethanolamine phosphate, or triethanolamine sulfate is used, but among these, in particular, sulfates such as triethanolamine lauryl sulfate and polyoxyethylene lauryl sulfate are used. Preferably, triethanolamine is used.

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

Moreover, the triethanolamine salt is 1 to 20% by weight, preferably 2 to 15% by weight, based on the solid content of the composition for detecting occult blood.
Preferably, it is present in weight percentages.

Buffer p++ The buffer is used to maintain a pH value near the pH at which the oxidizable indicator described above undergoes a favorable color change in the occult blood detection composition. The pl+buffer may be anything that can provide a predetermined pH value (for example, pH 5) to the composition for detecting occult blood, but specifically, it is possible to use a combination of citric acid and sodium citrate. preferable.

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

Polyester @ resin, alkyd resin, polyurethane resin, polystyrene resin, acrylic resin, 1-boxy resin, vinyl chloride resin, vinyl chloride copolymer resin, polyvinyl butyral resin, polyvinyl alcohol resin, polyvinyl pyrrolidone resin, or maleic anhydride copolymer Synthetic resins such as polymeric resins; natural polymers such as methylcellulose, ethylcellulose, hydroxyethylcellulose starch, polysaccharides, geladine, 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 at a ratio of 0 weight Φ%, preferably 0.5 to 1 weight 1%.

Water-absorbing powderWater-absorbing powder increases the water absorption of the test area formed by coating and drying on a support, promotes contact between the test liquid and the reagent composition, and enhances the color reaction of the support agent. It has a promoting function.

As such water-absorbing powder, it is not preferable to use one that shows extreme acidity or alkalinity when it comes into contact with water.
Moreover, those with high whiteness are preferable. Specifically, the following can be mentioned.

Kaolin, synthetic silica, glass, cellulose block,
Microcrystalline cellulose, ion exchange cellulose, ion exchange resin, calcium carbonate, t! Magnesium A acid, aluminum silicate, etc. can be used.

The water-absorbing powder has a content of 30 to 30% of the solid content of the composition for detecting occult blood.
It is desirable to use it in a proportion of 90% by weight.

The solvent is preferably one that can uniformly and stably dissolve or disperse each of the above components, especially the binder. Solvents that meet these conditions include: ■ aromatic hydrocarbons such as benzene and toluene, ■ aliphatic ketones such as methyl ethyl ketone, ■ esters such as ethyl acetate, and ■ non-alcohols such as n-butanol. An aqueous solvent or (1) water, or a mixture of (1) to (2) may be mentioned.

However, for the composition for detecting occult blood, it is preferable to use a non-aqueous solvent because the drying process after coating on the support can be carried out at a low temperature and in a short time. Use of a non-aqueous solvent also has the effect of preventing alteration and deterioration caused by residual components of the composition when applied onto the specimen.

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

Examples of 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. The wetting agent is 0.5 to 5% by weight based on the solid content of the composition for detecting occult blood.
It is preferable that the

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

In the microcapsule coating material that can be used in the present invention, 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, When divided into a second group consisting of a pH buffer, a water-absorbing powder, and a part of a binder, it is mixed with any of the components belonging to the first group or with any of the components belonging to the second group. An emulsifier that forms a good emulsion and quickly disintegrates when it comes into contact with water in the test liquid, and has the function of causing the contained organic hydroperoxide or oxidizable indicator, etc. to participate in the coloring reaction. It is desirable that

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

Polyvinyl alcohol resin, polyvinylpyrrolidone resin, polyacrylamide resin. or water-soluble synthetic resins such as polyvinyl acetate resin; natural polymers such as methylcellulose, ethylcellulose, hydroxycellulose, or carboxymethylcellulose starch, polysaccharides, gelatin, casein, gum arabic, or sodium alginate; In particular, gum arabic and carboxymethylcellulose vinyl alcohol resins are preferred.

The microcapsule coating material is preferably present in an amount of 2 to 70% by weight, preferably 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 obtain a test body.

Application methods include printing methods and coating methods (for example,
Roll coating method, spray coating method, dip coating method, solid coating method, etc. are used.

In the present invention, the amount of the composition for detecting occult blood to be applied is relatively large, and it is preferable that the amount of application be constant, so the silk screen printing method is used.

Preferably, the composition for detecting occult blood is applied onto the support by an intaglio printing method, a gravure printing method, or the like.

The amount of the composition for detecting occult blood applied onto the support varies depending on the type of composition, but is generally 2 to 150 g/Trt (when dry).
It is preferable that

It is preferable that the cross-sectional support be one that does not react with the composition for detecting occult blood and does not inhibit the coloring of the indicator.

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

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. Alternatively, the support itself may be in a form 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.

Hereinafter, the preparation of the composition for detecting occult blood and the production of test strips of the present invention will be described with reference to Examples.

Example 1 Among the compositions for detecting occult blood described below, cumene hydroperoxide and 6-medoxyquinoline, which is a sensitizer, were microencapsulated using gum arabic, and other components were added to form a composition for detecting occult blood. Prepared. Next, the composition for detecting occult blood was finely dispersed using a homomixer, and a square test reagent part with a side of 5 mm was printed on a white polystyrene sheet with a thickness of 300 μm using a screen printing method.
It was dried for 40 minutes at a temperature of .degree. The number of lines of the plate used for printing was 80 meshes, and the total thickness of the resist and screen gauze was 130 μm. After drying, it was cut into sticks of predetermined dimensions to obtain test specimens.

Composition for detecting occult blood Cumene hydroperoxide ... 3.6 parts by weight 6-medoxyquinoline 1.0 type assembly part (within the capsule) Gum arabic 9.4 double base part 3,
3',5.5'-tetramethylbenzidine...i. s11 parts citric acid...0.56 parts omega sl-lium citrate...2.2 parts lauryl sulfate triethanolamine...1,62 parts suspended polyethylene glycol...2.52 parts Flannel polyvinyl butyral ... 3.6-layer hanging section (Sekisui Chemical Co., Ltd., S-LEC BX-1) Cellulose fine powder ... 22.4-layer hanging section (Sekisui Chemical Co., Ltd., Avicel TG-D) n-amyl alcohol ・...39.7 parts by weight Butyl cellosolve acetate ...13.0 parts by weight As test liquid: - Normal urine - Urine containing 0.06 IRg/dl of human hemoglobin (manufactured by Sigma) - 0.151 Rg/dl of human hemoglobin Urine containing human hemoglobin (manufactured by Sigma) ■ Four types of urine containing human hemoglobin (manufactured by Sigma) of 0.75 ay/dl were prepared, and the test specimens obtained above were tested.
It was immersed in the liquid to be tested (3), taken out immediately after immersion, and allowed to stand for 1 minute, after which the color development of the test reagent portion was observed.

The coloration of the test reagent part is uniform and clear, and the color density increases stepwise as the human hemoglobin concentration in the test solution increases, and the concentration of occult blood in the test solution can be controlled within the above range. It was clearly distinguishable.

Even when the colored 1ζ test specimen was allowed to stand at room temperature for 5 minutes, no change in color tone was observed.

In addition, the obtained test specimen was sealed in a glass container,
After storage at 40°C for 12 months, a test was conducted in the same manner as above, and the coloration of the test reagent part was uniform, clear, and stable as before storage, and the human hemoglobin concentration in the test liquid 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, 20 red blood cells/μλ, 50 red blood cells/μ℃ in normal urine, and 250 red blood cells/μ℃ in normal urine were used. When the liquid to be tested was prepared and colored in the same manner, clear and stable spots appeared in the test reagent part, and the number of spots gradually increased as the concentration of red blood cells in the liquid to be tested increased. The concentration of red blood cells in the test liquid could be clearly determined within the above range. Even when the colored test specimen was allowed to stand at room temperature for 5 minutes, no change in color tone was observed.

In addition, the obtained test specimen was sealed in a glass container,
After storing it at 40°C for 12 months, we performed the same test as above, and found that the color of the test reagent part was as clear and stable as before storage, and the concentration of red blood cells in the test liquid was preserved. As before, it was possible to distinguish clearly.

Comparative Example 1 Test specimens were produced in the same manner as in Example 1, except that triethanolamine lauryl sulfate was not used and sodium lauryl'fiAWIi was used instead.

When the test liquid was tested in the same manner as used in Example 1, the coloration of the test reagent part was uniform, but the sensitivity and color density were low, and the test liquid in the low concentration range was It was difficult to clearly determine hemoglobin concentration.

Example 2 Same as Example 1, except that instead of 1.62 parts of triethanolamine lauryl sulfate, 1.13 parts of sodium lauryl sulfate and 0.73 part of triethanolamine hydrochloride were used.
When test specimens were produced in the same manner as in Example 1 using parts by weight, they exhibited similar coloration and had good storage stability.

Effect of K-excuse] In the present invention, since quinoline and triethanolamine salt are added to the composition for detecting occult blood, the sensitivity and coloring density of the composition for detecting occult blood can be improved, and it is possible to improve the sensitivity and color density of the composition for detecting occult blood. semi-quantification of hemoglobin as well as red blood cells. Furthermore, in the present invention, the occult blood detection area can be formed by a printing method rather than directly applying the occult blood detection composition on the support, which is advantageous for mass production and reduces the manufacturing process of the test body. Can be simplified.

Patent applicant: Dai Nippon Printing Co., Ltd. Agent Patent Attorney Katsu Ono Agent Patent Attorney Reiko Ohno Agent: Patent Attorney: Ryunosuke Ohno

Claims (1)

[Scope of Claims] 1. A composition for detecting occult blood that can give a detectable response in the presence of a peroxidizing active substance, which comprises a peroxide, an oxidizable indicator, quinoline or a derivative thereof, A composition for detecting occult blood, comprising a triethanolamine salt. 2. The composition for detecting occult blood according to claim 1, wherein the triethanolamine salt is triethanolamine lauryl sulfate or polyoxyethylene lauryl triethanolamine sulfate alone or in a mixture. 3. Claim 1, wherein the composition for detecting occult blood further comprises a pH buffer, a binding material, and a water-absorbing powder, each of which is dissolved or dispersed in a non-aqueous solvent. The composition for detecting occult blood according to section 1. 4. A composition for detecting occult blood capable of giving a detectable response in the presence of a peroxidizing active substance, comprising peroxide, an oxidizable indicator, quinoline or a derivative thereof, and a triethanolamine salt. A test object for detecting occult blood, characterized in that a composition for detecting occult blood is coated on a support. 5. The test object for detecting occult blood according to claim 4, wherein the triethanolamine salt is triethanolamine lauryl sulfate or polyoxyethylene lauryl triethanolamine sulfate alone or in a mixture. 6. Claim 4, characterized in that the composition for detecting occult blood further contains a pH buffer, a binder, and a water-absorbing powder, and each component is dissolved or dispersed in a non-aqueous solvent. A test specimen for detecting occult blood as described in .