JP2668705B2 - Inspection body - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a test body capable of simply and quickly detecting a specific component contained in a colored suspension such as blood, juice, urine, and industrial waste liquid. It is related to.

[Conventional technology]

In order to measure components in blood, it is necessary to separate serum or plasma from blood by membrane separation, centrifugation, etc. before testing, because the strong coloring of hemoglobin prevents all color reaction. However, such processing requires skill and considerable equipment. In order to prevent such a complicated operation and the disadvantage of erroneous recognition, Japanese Patent Publication No. 44-14673 has provided an absorbent support impregnated with a blood content detection reagent on a transparent support. A test material that separates plasma by covering the upper layer with a layer that separates blood cells and the like and performs a color reaction, and Japanese Patent Publication No. 45-15669 discloses that an absorbent carrier material containing a reagent for detecting blood contents is hydrophobic. A diagnostic material is described which flushes red blood cells which have become sexually deposited on the surface with water. Although these test materials have been improved in terms of simplification in use, they are not sufficient to provide semiquantitativeness in testing substances contained in blood.

Further, JP-A-59-228166 discloses a matrix having an upper layer and a lower layer, in which the upper layer contains a water-permeable polymer partially cross-linked and does not allow a colored dye to pass therethrough. Although it has been proposed that the sample of No. 1 is wiped off, the adsorption of the test sample components to the upper layer cannot be avoided in this case as well, and it is insufficient for the semi-quantitative test. Further, according to Japanese Patent Application Laid-Open No. 60-209174, an inspection device having a support layer and a microporous polymer layer containing a detection reagent is proposed. The color is determined by whether or not. Also in this case, the coloration of the test reagent is non-uniform, and the dye is adsorbed to the surface by wiping, and even if a transparent carrier is used, the deposited blood pigment is observed slightly, and the reagent color is observed. It may be an obstacle at times, and it is not enough to conduct a semi-quantitative test.

[Problems to be solved by the invention]

Since the test materials used for testing components in blood and urine in the above-described conventional techniques do not completely separate adsorption of hemoglobin, colored impurities, and the like contained in the test solution,
Their influence on coloration was unavoidable, and although they could be used for qualitative testing, they were not sufficient for semi-quantitative measurements. Further, as described above, in any of the test materials, blood is applied to the reagent portion or a portion in the vicinity of the reagent portion, so that the reagent leaks and diffuses, which also makes the semiquantitativeness of the test insufficient. .

The present invention solves the above-mentioned drawbacks of the conventional blood content test reagents, and tests not only blood but also pollutants including remarkably polluted urine, colored matters, fine suspended matters, etc., in industrial wastewater and the like. It is an object of the present invention to provide a simple inspection body capable of performing semi-quantitatively.

[Means for solving the problem]

The present invention relates to a test body for detecting and analyzing a test substance contained in a suspension containing a coloring substance, comprising a reagent layer provided on a support and capable of producing a response capable of detecting the test substance. An inspection including a coloring substance filterable coating layer which is releasably laminated on the reagent layer through an adhesive layer which is water-soluble and allows the components of the test body to pass therethrough, and which is easily peeled by immersion in a sample. It is related to the body.

[Action]

The present invention is characterized in that a test body is composed of a lower layer containing detection reagents provided on a support and an upper layer having a separation / filtration function which is removably laminated on the lower layer via an adhesive layer. The test body according to the present invention is immersed in the test solution for a certain period of time, or the test solution is dropped on the upper layer to allow only the solution containing the substance to be detected to pass through to the lower layer, and then peeled off by the water in the test solution. The upper layer, which has become easier, is peeled off, and the color of the lower test reagent layer is examined.

〔Example〕

 Hereinafter, the present invention will be described in detail.

As described above, the present invention is not limited to blood, and can be used for examination of urine with remarkably contaminated water, examination of suspension in a wide range including colored fine dispersions such as sewage and industrial wastewater, and adhesive dispersions. Things. First, examples of the substance capable of forming a film having a function of separating and filtering a colored fine dispersion, an adhesive dispersion, and the like, which are the upper layers constituting the test object, include the following.

 The materials contained in each type of film are listed.

Ion exchange membrane; cation exchange membrane, anion exchange membrane, amphoteric ion exchange membrane, bipolar ion exchange membrane, mosaic ion exchange membrane dialysis membrane: regenerated cellulose, cellulose acetate, polyacrylonitrile copolymer, polymethylmethacrylate, ethylene vinyl Alcohol copolymer, aromatic polysulfone, aromatic polyamide, carbonate, ethylene oxide copolymer reverse osmosis membrane; wholly aromatic polyamide, allyl-alkyl polyamide / polyureacellulose acetate, cellulose triacetate, polypiperazineamide ultrafiltration membrane; polyacryl Nitrile, polyvinylidene fluoride, polysulfone, polyethersulfone, aromatic nylon, cellulose acetate, Eval (Eval products, manufactured by Kuraray Co., Ltd.) Depending on the dispersion contained in the liquid to be detected from the above materials, one that does not hinder the permeation of the detection component is appropriately selected and used. For example, glucose in blood, urea nitrogen or industrial wastewater For the detection of low molecular weight substances such as iron ions, dialysis membranes such as cellulose membranes can be used to detect high molecular weight substances such as glutamate-pyruvate-transaminase and alkaline phosphatase in blood. A microfiltration membrane such as a polyvinylidene fluoride membrane filter can be used.

The upper layer and the lower layer are formed by immersing or contacting the test material in a test solution with an adhesive and then laminating the upper layer and the lower layer in close contact with each other with an adhesive so that the upper layer can be peeled off, or an adhesive layer therebetween. And glue them.

The adhesive layer adheres the upper layer and the lower layer over the entire surface or partially (spot shape or the like), and has a structure in which the inspection target substance and the solvent can reach the lower layer by dissolving or opening during the inspection.

The adhesive layer material is water-soluble and allows the sample substance to permeate, and it is easily peeled off by immersing it in the test liquid.The adhesive layer material is composed of the following, which may be used alone or as a mixture of two or more kinds. Used.

That is, acrylic resin, polyvinyl alcohol resin, polyvinylpyrrolidone resin, polyethylene oxide resin, maleic anhydride type copolymer resin, hydroxyethyl cellulose, carboxymethyl cellulose, polysaccharides, gelatin, sodium alginate can be mentioned.

Adhesion of the separation-filterable membrane to the support having the test reagent layer by the adhesive or the adhesive layer can be appropriately performed by a method such as heat sealing or dry lamination.

The lower test reagent layer is formed by impregnating a filter paper with a reagent composition, for example, as a test paper for detecting glucose in a body fluid, such as glucose oxidase, pen oxidase,
A reagent composition comprising an oxidizable indicator and a buffer is dissolved or dispersed in water containing gelatin or a water-alcohol solvent, and the obtained solution is impregnated with filter paper, and then dried to form a test reagent layer. Alternatively, it can be formed by appropriately cutting a filter paper and affixing it on a substrate such as a plastic film. Similarly, the reagent layer for protein detection and the reagent layer for pH detection may be impregnated with filter paper to manufacture a test reagent layer, which can be similarly formed.

Next, to provide a test reagent layer on a substrate by printing, enzymes are dissolved in advance in a water-alcohol mixed solution, and an indicator, a pH buffer, a polymer binder, a water-absorbing carrier, etc. are mixed with this. Then, an ink composition having printing or coating suitability is prepared, and the ink composition is printed (including coating) on a substrate and then dried to produce a test reagent layer. More preferably, as described in JP-A-62-263469 filed by the present applicant, (a) sugar oxidase, pen oxidase, oxidizable indicator, wetting agent, sensitivity regulator, stabilizer, pH buffer, binder,
And a reagent composition comprising a water-absorbing powder and a reagent composition dissolved or dispersed in a non-aqueous solvent, (b) an indicator showing a protein error, a pH buffer, a wetting agent, and a protein-adsorbing ion exchanger. An ink composition for detecting a protein in which a reagent composition comprising a form-retaining agent, a binder, and a water-absorbing powder is dissolved or dispersed in a solvent; (c) pH indicator, quaternary aluminum salt or amine salt, basic An ink composition for pH measurement, which is obtained by dissolving or dispersing a reagent composition comprising a substance, a binder, and a water-absorbing powder in a solvent, can be used by coating the ink composition on a substrate with a reagent. Layers can be formed. Further, for the detection of bilirubin in blood or urine, benzenediazonium salt having a substituent such as nitri, halogen, sulfone group, an adhesive, an acid (particularly a solid acid), a solvent, and if necessary a wetting agent, A filter paper is impregnated with the composition prepared by adding a water-absorbent carrier powder or the like, or printed or coated on a substrate. In addition, the reagent layer is formed using a known reagent for detecting body fluid such as a reagent for detecting glutamate-pyruvate-transaminase in blood.

The substrate is preferably one that does not react with the reagent composition and does not hinder the coloration of the reagent. Specifically, for example, paper, synthetic paper, non-woven fabric or synthetic resin film or paper and synthetic resin film And the like are used.

Techniques for applying the ink composition onto a substrate include printing methods,
A coating method (for example, roll coating, spray coating, dip coating, solid coating) or the like can be used. Since it is preferable that the coating amount of the ink composition is relatively large and the coating amount is constant, it is preferable to provide the ink composition on the substrate by a silk screen printing method, an intaglio printing method, a gravure printing method, or the like. . The coating amount varies depending on the type of the ink composition, but is generally preferably from ² to 150 g / m ² (when dried).

As the reagent layer that produces a response capable of detecting the test target substance as the lower layer, other than the above, known test strips for clinical tests used for detecting the test target substance, for example, commercially available blood, urine, etc. Test paper for detecting glucose in body fluid, test paper for detecting protein, test paper for detecting bilirubin, pH
Test papers such as a test paper for detection and a test paper for detecting glutamate-pyruvate-transaminase in blood can be used.

Also, the present invention provides a method for dissolving iron, hydrogen ions, biochemical oxygen demand (BOD), chemical oxygen demand (COD), chromium, and total mercury. ,
The present invention can be applied to the detection of copper and the like, and a commercially available ion test paper can be used as the reagent layer.

Example 1 Manufacture of a test piece for detecting blood sugar A dioxane solution of 10% nonionic linear polyethylene oxide (poly ox N-750; manufactured by Union Carbide, molecular weight 300,000) was bladed on 15 µ cellophane (# 250). Coat using a coater and dry to form an adhesive layer of about 10μ. On the other hand, after finely dispersing the glucose detection ink composition having the following composition with a homomixer, screen printing is performed so that a white polystyrene sheet having a thickness of 300 μ has a square shape with a side of 5 mm and a film thickness of 110 μ. did. The screen plate used was 80 mesh, and the total thickness of the resist and screen gauze was 130 μm.

The obtained printed matter was dried at 60 ° C. for 40 minutes to form a reagent layer.

Glucose detection ink composition Glucose oxidase (Toyobo Co., Ltd .; Grade II) 3.6 parts by weight Peroxidase (Toyobo Co., Ltd .; Grade III) 2.4 parts by weight Guayak butter 4.8 parts by weight Sorbitan monolaurate (Kao ( Span 20) 7.2 parts by weight Citric acid 2.8 parts by weight Sodium citrate 11.0 parts by weight Polyvinyl butyral (manufactured by Sekisui Chemical Co., Ltd .; trade name Esrec BX-1) 3.00 parts by weight Cellulose fine powder (Asahi Chemical Industry) Ltd .; Trade name Azecel
TG-D) 171 parts by weight n-amyl alcohol 228 parts by weight butyl cellosolve acetate 33.5 parts by weight Cellophane having an adhesive layer on the white polyethylene sheet having the reagent layer obtained as described above at a temperature of 80 ° C. of about 30 Heat sealing was performed by heating for 2 seconds, and this was cut into a stick to obtain a test piece for detecting blood sugar.

The drawing shows an example of the structure of the inspection body. Inspection body 1 is substrate 2
The reagent layer 3 provided above is provided so that the adhesive substance filterable coating layer 5 can be peeled off via the adhesive layer 4, and a tab piece 6 for facilitating the peeling operation can be formed. it can.
This test body was effective for semi-quantitative determination of blood glucose, and also had sufficient semi-quantitative ability for severe urine contaminated with blood, pus, and other foreign substances.

That is, about 0.2 CC of blood is dropped and applied onto the coating layer 5 of the test object 1 as shown in the drawing, left for about 1 minute, and then the coating layer 5 is peeled off and removed, and about 1 minute later, the blood is prepared in advance. As a result of comparison and measurement with a colorimetric table showing the coloring of each concentration, it was found that the glucose content in the blood to be tested was 50 mg / dl. On the other hand, when the coating layer 5 was not provided and blood was dropped directly on the reagent layer, the measurement was impossible due to the deposition of hemoglobin. Example 2 Manufacture of urea nitrogen detection test object In order to detect urea nitrogen in blood, the procedure was performed except that the following urea nitrogen detection ink composition was used instead of the glucose detection ink composition in Example 1. An inspection body was manufactured in the same manner as in Example 1.

Urea nitrogen detection ink composition Urease (Toyobo Co., Ltd .: Grade II) 3.6 parts by weight Peroxidase (Toyobo Co., Ltd .; Grade III) 2.4 parts by weight Guayak butter 4.8 parts by weight Sorbitan monolaurate (Kao Co., Ltd.) ); Product name span
20) 7.2 parts by weight Citric acid 2.0 parts by weight Sodium citrate 11.0 parts by weight Polyvinyl butyral (Sekisui Chemical Co., Ltd .; trade name Esrec BX-1) 3.00 parts by weight Cellulose fine powder (Asahi Chemical Industry Co., Ltd .; trade name Avicel) TG-D) 171 parts by weight n-amyl alcohol 228 parts by weight butyl cellosolve acetate 33.5 parts by weight About 0.2 CC of blood was dripped on the cellophane film of the obtained test sample, left for 1 minute after application, and the film was peeled off and removed. Observation of the sample left for 1 minute showed that the reagent layer was colored blue, and it was found that it contained about 10 mg / dl of urea nitrogen as compared with the standard colorimetric table.

Example 3 Production of Test Object for Total Cholesterol Detection In order to detect total cholesterol in blood, except that the following ink composition for detecting total cholesterol was used instead of the ink composition for detecting glucose in Example 1,
An inspection body was manufactured in the same manner as in Example 1.

Ink composition for detecting total cholesterol Cholesterol esterase (Toyobo Co., Ltd .; Grade
III) 1.6 parts by weight cholesterol oxidase (Toyobo Co., Ltd .; Grade
III) 1.6 parts by weight Peroxidase (Toyobo Co., Ltd .; Grade III) 2.4 parts by weight Guayak butter 4.8 parts by weight Sorbitan monolaurate (Kao Corporation; trade name: Span)
20) 7.2 parts by weight Monosodium phosphate 6.0 parts by weight Disodium phosphate 9.0 parts by weight Polyvinyl butyral (manufactured by Sekisui Chemical Co., Ltd .; trade name S-REC BX-1) 3.0 parts by weight Cellulose fine powder (manufactured by Asahi Kasei Co., Ltd.) ; Trade name: Azecel TG-D) 171 parts by weight n-Amyl alcohol 228 parts by weight Butyl cellosolve acetate 33.5 parts by weight About 0.2 CC of blood is dropped on the cellophane film of the obtained test sample, left for 3 minutes after application, and the film is peeled off When the sample was removed and left to stand for about 2 minutes, the reagent layer was colored blue, and it was found that the reagent layer contained about 200 mg / dl of total cholesterol when compared with the standard colorimetric table.

Example 4 Production of Glutamate-Pyruvate-Transaminase Detection Specimen Instead of the cellulose used in Example 1, a polyvinylidene fluoride membrane filter (trade name (MILLIPORE GVMP, manufactured by Millipore Company) was used, and a 10% hydroxyethylcellulose aqueous solution was used. Is coated to a thickness of 10 μm and dried, while a glutamate-pyruvate-transaminase detection ink composition having the following composition is finely dispersed with a homomixer and then screen-printed onto a white polyethylene sheet having a thickness of 300 μm. Printing was performed so that each side was a square having a side of 5 mm and the film thickness was 100 μm.The total thickness of the used screen plate 80 mesh, resist and screen gauze was 130 μm. It dried for a minute and formed the reagent layer.

Ink composition for detecting glutamate-pyruvate-transaminase Pyruvate oxidase (manufactured by Toyo Brewery Co., Ltd.) 4.0 parts by weight Pen oxidase (manufactured by Toyobo Co., Ltd .; Grade II) 0.5 parts by weight Guayak butter 4.8 parts by weight Alanine 3.2 parts by weight α-Ketoglutaric acid 3.0 parts by weight Tetraamine pyrophosphate 0.002 parts by weight Magnesium chloride 0.04 parts by weight Sorbitan Monolaurento (Kao Corporation; trade name Span 20) 7.2 parts by weight monosodium phosphate 9.38 parts by weight disodium phosphate 4.54 Parts by weight Polyvinyl butyral (Sekisui Chemical Co., Ltd .; trade name: SREC BX-1) 3.00 parts by weight Cellulose fine powder (manufactured by Asahi Kasei Corporation; trade name: Avicel SF) 171 parts by weight n-amyl alcohol 228 parts by weight butyl cellosolve acetate 33.5 parts by weight White polyethylene with reagent layer obtained as above The membrane filter having an adhesive layer laminated on the sheet, subjected to heat-sealing by heating at a temperature of 80 ° C. for about 1 minute to give a test strip and cut into sticks.

About 0.2 CC of blood was dropped and applied on the membrane filter of the test piece, left for about 3 minutes, the membrane filter was peeled off and removed, and the color of the reagent layer was observed when the color of the reagent layer was observed for about 2 minutes. , Approximately 20 IU / much of glutamate-pyruvate-transaminase activity was estimated.

Example 5 Production of Fe (II) Detection Specimen A dioxane solution of 10% nonionic linear polyethylene oxide (poly ox N-750; manufactured by Union Carbide, molecular weight 300,000) on 15μ cellophane (# 250). Is coated with a blade coater and dried to form an adhesive layer of about 10 μm. On the other hand, after the Fe (II) detection ink composition having the following composition is finely dispersed by a homomixer, a white polystyrene sheet having a thickness of 300μ is formed into a square having a side of 5 mm and a film thickness of 110μ by finely dispersing with a homomixer. As printed. The screen plate used was 80 mesh, and the total thickness of the resist and screen gauze was 130 μm.

The obtained printed matter was dried at 60 ° C. for 40 minutes to form a reagent layer.

Fe (II) detection ink composition o-phenanthroline 0.5 parts by weight Sorbitan monolaurate (manufactured by Kao Corporation; trade name: Span 20) 7.2 parts by weight Aluminum acetate 15.0 parts by weight polyvinyl butyral (manufactured by Sekisui Chemical Co., Ltd .; Trade name SREC BX-1) 3.0 parts by weight Cellulose fine powder (Asahi Kasei Kogyo Co., Ltd .; trade name azecel TG-D) 171 parts by weight n-amyl alcohol 228 parts by weight Butyl cellosolve acetate 33.5 parts by weight Formed as described above. A cellophane having an adhesive layer was adhered on a white polyethylene sheet having a reagent layer by heating at a temperature of 80 ° C. for about 30 seconds to produce a test piece.

When measured with a turbidimeter, 5 mg of Fe ²⁺ showing the turbidity of DO 300
/ dl to and Fe ²⁺ with industrial wastewater containing 100 mg / dl, respectively, were examined for Fe ²⁺ with Fe (II) for detecting the inspection body obtained by the above. After immersing the test body in industrial wastewater, the cellophane, which is the coating layer, was peeled off and observed, and the coloration of the reagent layer was observed. As a result, it was possible to determine the Fe ²⁺ concentration of the above two kinds of liquids. However, when the test body without the cellophane coating on the reagent layer was similarly immersed in industrial wastewater, the surface of the test body was markedly colored and it was impossible to distinguish the liquids with different Fe ²⁺ concentrations. Met.

〔The invention's effect〕

The present invention relates to a test body in which a coloring substance filterable coating is releasably provided on a reagent layer provided on a support and capable of producing a response capable of detecting a detection target substance. In a simple test for a colored test liquid such as wastewater, the colorant filterable film layer on which the colorant is attached can be easily peeled off, and the color of the reagent layer can be confirmed. Further, it is possible to suppress the diffusion of the reagent on the reagent layer into the test liquid. Therefore, the test body according to the present invention was able to semi-quantitatively detect the components contained in the coloring liquid. Furthermore, since the test body of the present invention is provided with a reagent layer via an adhesive layer which is securely held until the test layer is immersed in the test solution and is easily peeled off after immersion in the test solution, , Manufacturing,
It is convenient for handling during the process from transportation to use.

[Brief description of the drawings]

The drawing is an enlarged sectional view showing an example of the test object according to the present invention. 1 ... Inspection body, 2 ... Substrate, 3 ... Reagent layer, 4 ... Adhesive layer, 5 ... Coloring substance filterable coating layer, 6 ... Tab piece

Claims (1)

(57) [Claims] 1. A test body for detecting and analyzing a test target substance contained in a suspension containing a coloring substance, which can generate a response capable of detecting the test target substance provided on a support. A reagent layer and a group consisting of acrylic resin, polyvinyl alcohol resin, polyvinylpyrrolidone resin, polyethylene oxide resin, maleic anhydride copolymer resin, hydroxyethyl cellulose, carboxymethyl cellulose, polysaccharide, gelatin, and sodium alginate on the reagent layer. An inspection body comprising a coloring substance filterable coating layer which is releasably laminated via an adhesive layer made of one or more selected substances.