JPS63241355A - Composition for detection of urobilinogen and object to be inspected using the same - Google Patents

Abstract

PURPOSE:To reduce reaction time along with clear coloration, by adding an anionic surfactant or non-ionic surfactant as reaction activating agent in a urobilinogen detection composition using an Ehrlich's reagent. CONSTITUTION:An Ehrlich's reagent such as p-dimethyl aminobenzaldehyde, a strong-acid buffer such as sulfosalicylate, a binder such as polyester resin, a water absorbing carrier powder such as microcrystal cellulose, a non-water solvent such as ester and a reaction activating agent comprising an anionic surfactant or non-ionic surfactant are finely dispersed with a high-speed agitator. The composition thus obtained is screen printed on a white polystyrene sheet to make a test piece by drying. The test piece is immersed into a liquid to be inspected, taken out immediately to be left along for 60sec and then, coloring is observed.

Description

Detailed Description of the Invention [Field of Industrial Application] The present invention relates to a detection composition for detecting urobilinogen in body fluids, particularly urine, and a test body formed using the detection composition. It is something.

[Conventional technology]

Urobilinogen concentration, such as in urine, is an important indicator for diagnosing liver and gallbladder diseases.

Conventionally, as a method for detecting urobilinogen, p
- There is a method using the so-called "Ehrlich reaction" using dimethylaminobenzaldehyde or the like as an indicator, in which such an indicator is immersed in a water-absorbing carrier together with a strongly acidic buffer, dried, and then transferred to another support. It is pasted on top to serve as a test object and used to detect urobilinogen. Although such a test object has the advantage that it can be operated using an hour wheel and the determination can be made in a short time, it has the problem that the manufacturing process is complicated and it is not suitable for mass production.

As a urobilinogen test material suitable for mass production, one has been proposed that is obtained by pattern-printing an ink composition containing an indicator, a strong acidic buffer, a binder, and a water-absorbing carrier powder onto a support, and drying the pattern. (Unexamined Japanese Patent Publication No. 60-178
However, this test specimen has a problem in that the reaction rate is slow, and the difference in coloration depending on the concentration of urobilinogen is not clear, making it difficult to determine the concentration.

[Problem that the invention seeks to solve]

The present invention aims to solve the problem that hitherto there has been no test specimen suitable for mass production, requiring a short reaction time, and with clear differences in coloration depending on the concentration of urobilinogen. .

[Means for solving problems]

As a result of the research conducted by the present inventors, we have found that by further adding a reaction activator to the specimen using the Ehrlich reaction, and in particular adding an anionic surfactant and a nonionic surfactant as a reaction activator, the above It has become clear that the problems in the prior art can be solved, and the present invention has thus been completed.

The present invention relates to "a composition for detecting urobilinogen, characterized in that an Ehrlich reagent, a strong acidic buffer, a binding material, a water-absorbing carrier powder, and a reaction activator are dissolved or dispersed in a non-aqueous solvent" and urobilinogen test material J, characterized in that a composition comprising r Ehrlich's reagent, a strong acidic buffer, a binder, a water-absorbing carrier powder, and a reaction activator is coated on a support. It is something.

[Effect]

According to the present inventors, cationic surfactants suppress the Ehrlich reaction, whereas anionic surfactants promote the reaction between the test specimen and urobilinogen, and nonionic surfactants Among them, HLB is 6.0 to 11.0
A substance within this range improves the contact between the specimen and urobilinogen.

(Example) Below, the composition for detecting urobilinogen and the urobilinogen test specimen of the present invention will be explained in order.

+a+Principle When p-di(alkyl)-7 minobenzaldehyde, an indicator, comes into contact with urobilinogen while maintained at an acidic pH, it forms a complex, which produces a yellow-pink color depending on the concentration of urobilinogen present in the test liquid. - It exhibits a reddish-purple color, and this principle makes it possible to detect urobilinogen in the test liquid.

Ehrlich reagent (indicator) Specific Ehrlich reagents are p-di(lower alkyl)aminobenzaldehydes such as p-dimethylaminobenzaldehyde and p-diethylaminobenzaldehyde. Among these, p-diethylaminobenzaldehyde is superior in terms of sensitivity.

Ehrlich's reagent is preferably 0.0 in the detection composition.
It is used at a rate of 1 to 5%.

fe1 Strongly acidic buffer Since the reaction between urobilinogen and the indicator proceeds in an acidic region, in order to provide such a reaction environment and to suppress side reactions, a buffer that can adjust the pH to 3 or higher, preferably solid at room temperature, is used. acid. Specifically, sulfosalicylic acid, oxalic acid, p-toluenesulfonic acid, metaphosphoric acid, etc. are used alone or in combination.

The strong acidic buffer is preferably used in the detection composition in an amount of 2 to 30%.
Used in proportions by weight.

+d+Binding material The binding material is required to be one that does not affect the components and pH of the test liquid, does not affect the reagents, especially the indicator, and does not interfere with the color reaction. . The following are examples of binding materials that have been confirmed to satisfy these conditions.

(1) Synthetic resin 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, polyvinyl pyrrolidone resin, or maleic anhydride system copolymer resins, etc.

(2) Cellulose derivatives methylcellulose, ethylcellulose, hydroxyethylcellulose, carboxymethylcellulose, etc.

(3) Natural polymer starch, polyPR, gelatin, casein, or sodium alginate.

The binder is preferably present in the detection composition at a concentration of 0. It is desirable to use it in a proportion of 2 to 20% by weight.

(e) Water-absorbing carrier powder The water-absorbing carrier powder has the function of promoting contact between the test liquid and the indicator and promoting the color reaction of the indicator.

Such a water-absorbing carrier powder is preferably one that exhibits extreme acidity or alkalinity when it comes into contact with water, and preferably one that exhibits high whiteness. Specifically, the following can be mentioned.

Kaolin, synthetic silica, glass, cellulose block,
Microcrystalline cellulose, ion exchange cellulose, ion exchange resin, magnesium carbonate/aluminum silicate, etc. can be used.

The water-absorbing carrier powder preferably contains 15 to 15% in the detection composition.
It is preferable to use it in a proportion of 50% by weight.

(f)? As the vehicle solvent, it is preferable to use one that can uniformly and stably dissolve or disperse the above (bl to tel components, especially the binder).Solvents that meet these conditions include aromatic hydrocarbons, esters, etc. and non-aqueous solvents such as alcohols, water, or mixtures thereof.

However, as a composition for detecting urobilinogen, 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. Another effect is that it is possible to prevent deterioration in quality caused by residual moisture in the composition that has been applied onto the specimen and dried.

The solvent is preferably 30 to 85% by weight in the detection composition.
used in amounts of

(Bright reaction activator As the reaction activator, an anionic surfactant or a nonionic surfactant is used, and these are preferably used in combination.

Examples of anionic surfactants include sodium lauryl sulfate and sodium dodecyl sulfonate, which are used alone or in combination in the yokode composition preferably in the 11.0 range. Specifically, in the detection composition, preferably 0.05 It is used in a proportion of ~5% by weight.

(h) Other components In addition to the above-mentioned main components, the composition of the present invention can contain many auxiliary components. For example, the color sensitivity can be increased by adding caffeine, and technically acceptable ingredients such as antioxidants and chelating agents can be included. Further, a background dye may be added for the purpose of making the color tone of the indicator easier to see.

(M) Preparation of Composition for Detecting Urobilinogen For each of the above-mentioned components, water-absorbing carrier powder is first ground into particles having a diameter of 50 μm or less. The obtained particles are then treated with an indicator,
A strongly acidic buffer, a binder, and a reaction activator may be added to a solution in which they have been previously dissolved or dispersed in a solvent, and then dispersed and mixed using a high-speed stirrer, sand mill, ball mill, Miki roll, ultrasonic disperser, etc. .

('') Manufacture of test object The composition for detecting urobilinogen as described above is applied as an ink composition onto a support, and then dried to form a region for detecting urobilinogen, and the test object is can get.

Application methods include printing method, coating method (e.g. Eva,
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 urobilinogen detection is relatively large, and it is preferable that the coating amount is constant. Preferably, the composition is coated onto a support.

The amount of the composition for detecting urobilinogen applied onto the support varies depending on the type of the composition, but -a is 2 to 150 g/n?
(when dry) is preferable.

The support is preferably one that does not react with the composition for detecting urobilinogen and does not inhibit the coloring of the indicator.

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

The test body in which the urobilinogen detection area is formed on such a support can be in the form of a stick, roll, tape, or
It may be formed in the form of. Alternatively, the support itself may be in a form in which the test liquid can be collected, for example, in the form of a pot,
It may be formed into a test tube shape, a dish shape, a tray shape, or a dropper shape.

〔Effect of the invention〕

The composition for detecting urobilinogen of the present invention includes an anionic surfactant as a reaction activator and an HLB of 6.0 to 6.0.
Since the nonionic surfactant in the range of 11.0 is used, the reaction rate is increased compared to conventional ones, resulting in shorter color development time and increased color sensitivity.
Furthermore, fading of color over time is suppressed, making it useful for clinical tests that require simplicity and speed.

[Specific examples]

After preparing a composition for detecting urobilinogen having the following composition by finely dispersing it with a high-speed stirrer, a square with one side of 51IIIO was printed on a white polystyrene sheet with a thickness of 300μm using a screen printing method.
It was dried for 30 minutes at a temperature of 'C. The number of lines of the plate used for printing was 8o mesh, and the total thickness of the resist and screen gauze was 180 μm. After drying, it was cut into predetermined dimensions to obtain test specimens.

p-diethylaminobenzaldehyde for urobilinogen--111----------1-
・2.45 parts by weight sulfosalicylic acid ---
-1--...------1...-125 parts by weight Sodium laurate--2, 40M 1 part Sorbitan monolaurate (HLB8.6)-- −
・−・・−・−・−・・・・−1,3 parts by weight Caffeine−・・・−・・−・−・−・・−
・・−・−・・−−−11−−−One song 16 mfft part Methyl vinyl ether/maleic anhydride copolymer (GA
Manufactured by Company F, Gantorenz AN-169) amyl alcohol hafestel
−・−・・−・−−−−1・−・−・−・・7
5 parts by weight ethylene glycol monomethyl ether...
−〜・・−・・−−−−−−・・−・・−・−・・
-421N part ethylene glycol monomethyl ether acetate-・-・・・・・・−・−−−−1−・−
・・・・・・・・・・・・・−・−125 parts Sodium carboxymethylcellulose−−−−−−−−−−−
−・・−・・−・−・−・・・・・・・−・3
1 part microcrystalline cellulose (manufactured by Fukaisei Co., Ltd., Avicel TG
−D )−−−・・・・−・−・−−−−−−−−・−
--1--・--1--・156 parts by weight As test liquid, ■ Normal urine ■ I EU/att Urobilinogen-containing urine ■ 5
Three types of EU/d6 urobilinogen-containing urine (EU means Ehrlich unit) were prepared, and the test specimens obtained above were
Immerse it in the liquid to be tested, and take it out immediately after immersion.
The color development after standing for a second was observed, and the following results were obtained.

■・・・・・・－・・・Pale yellow ■－・・・－・・・－・Pale pink ■－・・・－・－Red-orange Moreover, the above coloration did not change even after 120 seconds.

! JLfLL In the composition for detecting urobilinogen prepared in Example 1,
Sorbitan monooleate (HLB4.3) and polyoxyethylene (2
HLI
39.0 ink was prepared, a test piece for inspection was produced, and the test pieces were colored using test liquids ① to ②.The results were as follows.

■------------Pale yellow ■－・・・・・・－・Pale pink ■－・・・・・・－Red-orange Moreover, the above coloration did not change even after 120 seconds.

Upper J Enkan 11 In the composition for detecting urobilinogen prepared in Example 1,
Sorbitan monooleate (HLB4.3) and polyoxyethylene (
20) Using a mixture of sorbitan monooleate (HLB15.O) and using the same method as in Example I, HL
B5.0 ink was prepared, test pieces were produced, and colored using test liquids ① to ②, and the results were as follows.

■・・−・・・Pale yellow (unchanged) ■−・・−・・
...Pale yellow (light yellow-pink)■...--Pale yellow-pink (dark red-orange) Also, after 120 seconds, the color of those in which urobilinogen was detected changed.

The numbers in parentheses () to () indicate the color after 120 seconds.

In the composition of Example 1, only sorbitan monolaurate was removed from the composition for detecting urobilinogen. The coloring was carried out in the same manner as in Example 1, and the results were as follows.

■・・−・−・−Pale yellow (unchanged) ■・−・・−・・Pale yellow (light yellow pink) ■・・−・・−・−・Pale yellow pink (light pink) Also, 120
After seconds, those that detected urobilinogen changed color.

The color in parentheses for Φ to ■ indicates the color after 120 seconds.

Upper plate ■↓ A composition for detecting urobilinogen was prepared by removing only sodium lauryl sulfate from the composition of Example 1, and coloring was performed in the same manner as in Example 1. The results were as follows.

■・−・−・−・Pale yellow (unchanged) ■・−・−・
...-Pale yellow-pink (pale pink) -...-Dark red-orange (red-orange) Also, after 120 seconds, the color of those in which urobilinogen was detected changed.

The numbers in parentheses () to () indicate the color after 120 seconds.

Procedures Amendment Spontaneous writing) December 22, 1985 1. Indication of the case Patent Application No. 75677 of 1988 2. Title of the invention: Composition for detecting urobilinogen and specimen using the same 3. Person making the amendment Relationship to the incident Patent applicant address Sumire no Kouhai Trade flute yIO Hiopai Shin 1 port 1 Me name (289) Ryunashi Kinuta k”j Kosmanbashira representative 1r Nezumi So ryo 4, agent 〒162 Address Tokyo 1-1, Ichigaya Kagacho-chome, Shinjuku-ku, Tokyo (voluntary) 6. Number of inventions increased by amendment Tl) The scope of claims will be amended as shown in the attached sheet.

(2) Amend "more than" in the bottom line of page 6 of the specification to "r or less".

(3) [Benzene] is inserted between "dodecyl" and "sulfonic acid" on page 1, line 7 of the specification.

Claims (1) Ehrlich reagent, strong acidic buffer, binding material,
A composition for detecting urobilinogen, characterized in that a water-absorbing carrier powder and a reaction activator are dissolved or dispersed in a non-aqueous solvent.

(2) The reaction activator is an anionic surfactant and H
The composition for detecting urobilinogen according to claim (1), which is a nonionic surfactant having an LB of 6.0 to 11.0.

(3) The composition for detecting urobilinogen according to claim (2), wherein the anionic surfactant is sodium lauryl sulfate or sodium dodecylbenzenesulfonate.

(4) Ehrlich reagent, strong acidic buffer, binding material,
1. A urobilinogen test material, characterized in that a composition comprising a water-absorbing carrier powder and a reaction activator is coated on a support.

(5) The reaction activator is an anionic surfactant and H
The urobilinogen test material according to claim (4), which is a nonionic surfactant with an LB of 6.0 to 11.0.

(6) The urobilinogen test material according to claim (5), wherein the anionic surfactant is sodium lauryl sulfate or sodium dodecylbenzenesulfonate.

Claims (6)

[Claims] (1) A composition for detecting urobilinogen, characterized in that an Ehrlich reagent, a strong acidic buffer, a binder, a water-absorbing carrier powder, and a reaction activator are dissolved or dispersed in a non-aqueous solvent. (2) The reaction activator is an anionic surfactant and HLB
The composition for detecting urobilinogen according to claim (1), which is a nonionic surfactant with a surfactant of 6.0 to 11.0. (3) The composition for detecting urobilinogen according to claim (2), wherein the anionic surfactant is sodium lauryl sulfate or sodium dodecylbenzenesulfonate. (4) A urobilinogen test material, characterized in that a composition comprising Ehrlich's reagent, a strong acidic buffer, a binder, a water-absorbing carrier powder, and a reaction activator is coated on a support. (5) The reaction activator is an anionic surfactant and HLB
The urobilinogen test material according to claim (4), which is a nonionic surfactant with a molecular weight of 6.0 to 11.0. (6) The urobilinogen test material according to claim (5), wherein the anionic surfactant is sodium lauryl sulfate or sodium dodecyl sulfonate.