JPS63221243A - Composition for testing and test piece - Google Patents

Abstract

PURPOSE:To permit quick and exact detection of urobilinogen in bodily fluid by using a specific compd. as a diazo compd. to be used in detection of the urobilinogen based on a diazo reaction. CONSTITUTION:A compsn. contg. at least one of the diazo compds. expressed by the general formulas I-IV is used as the compsn. for testing to detect the urobilinogen, more particularly, the urobilinogen in urine. This compsn. is capable of deciding the concn. of the urobilinogen in urine in a short period without being affected by extraneous matter such as bilirubin co-existing therein. Since there is a large difference in the density of the coloration arising from the various concns. of the urobilinogen, the sure detection of the urobilinogen is permitted.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention relates to a test composition and a test piece for detecting urobilinogen, particularly in body fluids, especially urine.

[Conventional technology]

The concentration of urobilinogen in urine is an important indicator for diagnosing the presence or absence of liver II and gallbladder diseases.

As a method for detecting this urobilinogen, a method utilizing the "Ehrlich reaction" using a hydrochloric acid solution of p-dimethylaminobenzaldehyde is known as a standard method.

Test strips utilizing the above-mentioned "Ehrlich reaction" are also used in clinical tests that require simplicity and speed.

However, the above-mentioned conventional test strips have poor specificity in their reaction with urobilinogen, i.e., indole, skatole, and vorphobilinogen present in the urine are excreted after the administered drug is excreted or metabolized. Because it undergoes a significant reaction with aromatic amines and, in particular, urea, which is relatively present in urine, the inherent reaction of urobilinogen is hindered, and the reaction rate of the coloring reaction is slow, making it difficult to judge. It has the disadvantage of having to wait for at least one minute before the process.

Therefore, in recent years, attempts have been reported to use test pieces that utilize diazo reactions using various aromatic diazonium salts (for example, JP-A-48-11093, JP-A-49-1999).
44797),.

When using test pieces that utilize these diazo reactions,
Compared to using a test piece that uses the Ehrlich reaction, it has the advantage of a higher reaction rate with urobilinogen, but on the other hand, it also reacts with coexisting bilirubin, etc.
There is a drawback that the determination is inaccurate, and furthermore, the difference in color density due to the high and low urobilinogen concentrations is an unknown value, making it difficult to determine the density.

[Problem that the invention seeks to solve]

Therefore, the present invention aims to solve the problems that conventional test pieces lack specificity in their reaction with urobilinogen, and that the difference in coloration depending on the concentration of urobilinogen is unclear. be.

[Means for solving problems]

In this invention, the diazo compound used in the detection of urobilinogen is based on the diazo reaction.
It has become possible to solve the above problems by using the following specific items.

That is, this invention has the following formula: The present invention relates to a test composition for detecting urobilinogen, which is characterized by containing at least one of the compounds represented by the following, and a test piece using the test composition.

〔Example〕

(Diazo Compound) In the diazo compound represented by the general formulas [1] to (4) used in this invention, the phenylene group represented by pH in the formula includes, in addition to unsubstituted ones, alkyl groups, There are phenylene groups substituted with alkoxy groups, halogen atoms, etc.

Furthermore, the stabilizing anions represented by (X)- and (Y)' in the diazo compounds represented by general formulas [1] to [4] include sulfate ion, tetrafluoroborate ion, trifluoroborate ion, and trifluoroborate ion. Examples include fluoromethylsulfonate ion and arylsulfonate ion.

Among these, those using tetrafluoroborate ions are preferable in terms of the storage stability of the test composition.

In the above, (X)'' and (Y)- may be the same or different, and when used in a test composition, each of the groups [1] to [4] above Compounds within the same group may be combined, or compounds from other groups may be combined.

The compounds (1) to [4] above can be prepared by known methods in diazo chemistry. Alternatively, they can be prepared in solution or dispersion from the corresponding amines by known methods known in diazo chemistry.

(Composition) The above diazo compounds include adhesives, acids (especially solid acids)
After adding a solvent and, if necessary, a wetting agent, a water-absorbing carrier powder, etc. to form a composition, it is impregnated or coated onto a support, and then heated at the lowest possible temperature to suppress the decomposition of the diazonium salt. It can be dried and used as a test piece.

(Adhesive) The adhesive (binder) is one that does not affect the components and hydrogen ion concentration index in the test liquid, does not affect the reagents, and does not interfere with the color reaction. This is required.

Adhesives that have been confirmed to meet these conditions include: (1) Synthetic resins such as polyester resins, alkyd resins, polyurethane resins, acrylic resins, epoxy resins, vinyl chloride resins, and vinyl chloride copolymer resins (e.g., vinyl chloride /vinyl acetate copolymer resin), polyvinyl butyral resin, polyvinyl alcohol resin, polyvinyl pyrrolidone resin, or maleic anhydride copolymer resin, etc., ■Cellulose derivatives such as methylcellulose, ethylcellulose, hydroxyethylcellulose, or carboxymethylcellulose, or , ■ Natural polymers include starch, polyamide, gelatin, casein, and sodium alginate.

(Acid) It is necessary to add a sufficient amount of acid for the reaction between urobilinogen and the diazo compound, and preferably an acid that is solid at room temperature, such as citric acid, oxalic acid, sulfosalicylic acid, p-toluenesulfone. Acid or metaphosphoric acid may be used alone or in combination.

(Wetting agent) A wetting agent helps in dispersing each reagent, promotes formation of a uniform reagent layer, and can improve water wettability.

As the wetting agent, for example, surfactants such as nonionic surfactants, anionic surfactants, cationic surfactants, or amphoteric surfactants, or polyethylene glycols can be used.

(Water-absorbing carrier powder) By blending the water-absorbing carrier powder into the test composition, it has the function of promoting contact between the test liquid and each reagent and promoting color reaction.

Such a water-absorbing carrier powder is preferably one that exhibits extremely acidity or alkalinity when it comes into contact with water (moreover, one that exhibits high whiteness is preferable).

Specific water-absorbing carrier powders include kaolin, synthetic silica, glass, cellulose block, microcrystalline cellulose, ion exchange cellulose, ion curing resin, calcium carbonate, aluminum silicate, and the like.

(Solvent) The solvent is preferably one that can dissolve or disperse the above components.Solvents that satisfy this condition include non-aqueous solvents such as alcohols, esters, aliphatic hydrocarbons, aromatic hydrocarbons, etc. or water, or a mixture thereof.

(Blending ratio) In the above test composition, the mixing ratio of each component is preferably as follows.

Diazo compound・・・・・・・・−0゜002〜0.3
Weight% Adhesive・・・・−・−・・・・・−・−・
・−・−−−−0, 5~1O wt% acid ・・−・−
...
・-・・-・−・−・・2～30wt% wetting agent・
・−・−・・−・−・・〜・−・−・・−・・
・-・-0.2~10% by weight water-absorbing bundle ・-・-・
・・・−・−・−・・−・・・20 to 50% by weight solvent・・・・−・−・・−・・−・−・・・・・・・・・・
-································································40 to 95% by weight (manufacture of test pieces) As one method, the above test composition is prepared by using filter paper, polyester fleece, It is impregnated into an absorbent substrate such as a plastic sheet, dried, and then tested either as it is or by attaching it to another support, such as a plastic sheet such as a polystyrene sheet or a polyvinyl chloride sheet, using double-sided tape. There is a way to separate it.

Alternatively, instead of impregnation, the above composition may be printed or applied onto a suitable non-absorbent support such as synthetic paper, synthetic resin film, laminate of paper and synthetic resin film, etc. The specimen may be coated, dried, and used as it is or by being attached to another support.The other support is the same as the support used for printing or coating. can be used. As the printing or 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 this invention, among the two methods broadly classified above, the latter method, especially the printing method, allows a relatively large amount of the test composition to be coated, and the amount of the test composition to be coated is constant. Therefore, silk screen printing method, intaglio printing method,
Preferably, the test composition is provided on a support by a gravure printing method or the like.

The amount applied during printing or coating varies depending on the type of test composition, but is generally preferably about 2 to 150 g/rrf (dry).

The test piece thus obtained was immersed in the test liquid so that the test composition came into contact with it, and the color tone obtained was obtained by a color reaction with a liquid having a known urobilinogen concentration. The urobilinogen concentration in the liquid to be tested can be determined by comparing it with the standard color table after a predetermined period of time after immersion.

Although a test piece is a preferred form of use for the test composition of the present invention, it does not have to be in the form of a test piece, but can be used in the form of a solution, brought into contact with the liquid to be tested to develop color, and prepared in advance. It goes without saying that it is also possible to quantify urobilinogen using a standard color chart or using a spectrophotometer. In this case, the resulting pigment is extracted into an organic solvent such as chloroform and a chromaticity chart It is preferable to compare or measure absorbance.

[Effect]

According to this invention, the above-mentioned diazo compounds (1) to [4] (individually or as a mixture) react almost instantaneously with urobilinogen in the presence of an acid to exhibit a reddish-orange color. The above diazo compound does not react with impurities such as urea, aminosalicylic acid, or sulfonamide, and
The reactivity with indole, skatole, and vorphobilinogen is significantly lower than that with urobilinogen.
Even when large amounts of bilirubin, which often coexist, are present, the reactivity with bilirubin is significantly lower than that with urobilinogen, so the reaction with bilirubin proceeds after the reaction with urobilinogen has substantially completed. Therefore, the possibility of interfering with the detection of urobilinogen is extremely low.

〔Effect of the invention〕

The test composition of the present invention allows the concentration of urobilinogen in a test liquid to be determined in a short time without being affected by contaminants such as coexisting bilirubin, and color changes depending on the concentration of urobilinogen. Since the difference in concentration is large, urobilinogen can be reliably detected in test fluids, especially body fluids, and especially urine, and is extremely useful for clinical tests that require simplicity and speed.

[Specific examples]

A test composition with the following composition was prepared, and filter paper (Toyo Roshi type,
No. 51 was impregnated and dried at a temperature of 45° C. for 40 minutes to produce a test piece.

4.4'-bis-diazonium diphenylene sulfone sulfate--0,02 parts by weight of sulfosalicylic acid-1-- −・−１−−−−・・
················································································ 8 parts by weight 0.4 parts by weight methyl alcohol・・・・・・−・−・−・−・−・
−・・・・～・−20 weight 1 part water・−・−・−・・−
・・−・−・・−・−・−・・・・−・・・・−・
−・・−・・−・−・・−・−・−・So heavy weight part of test liquid, ■; normal urine, ■; 1 g/d 1 urobilinogen-containing bundle, ■; 5 mg Urine containing /d6 of urobilinogen and ■: bundle containing bilirubin of 5 g/dj were used.The test piece obtained above was immersed in each of the test liquids, immediately taken out, and After standing for a second, color development was observed and the following results were obtained.

■−・−・・−・No discoloration.

■−・・−・・・Pale pink coloration.

■－・・・－・・・－Pink coloration.

■－・・・－No discoloration.

11 and 1 The same procedure as Example 1 was carried out except that a test composition having the following composition was used.

4.4° -Bis-diazonium benzophenone aryl sulfonate --- 0.02 parts by weight Metaphosphoric acid --- ---・・・
・・−・・−・・−１−−−−−・−・−・−
・−・・−・6 parts by weight Sodium dodecyl sulfate・−・・
・−・・・・・−・0.6 parts by weight Methyl alcohol
・・・−・1s weight 1 part water・−・−・−・−・・−
・−・−・−・・−・−・−・・−・・・・・・・・・・・−
......85 parts by weight The color development of the test liquid in the same manner as in Example 1 was as follows.

■・・・・・・・・・－・－No discoloration.

■−・・−・−・Pale pink coloration.

■・・・－・－・・・Pink coloration.

■・−・−・・−・No discoloration.

11 Portions 11 A test composition having the following composition was prepared by finely dispersing it using a homomixer, and then printed in a rectangular shape with a size of 5@11×5■■ on a 300 μm thick polystyrene sheet by screen printing.

After printing, it was dried at a temperature of 45° C. for 40 minutes and cut into a stain shape to obtain a test piece. The number of lines of the screen plate used for printing was 80 meshes, and the total thickness of the resist and screen gauze was 160 μm.

4.4° -Bis-diazonium diphenyl sulfide tetrafluoroborate・・・・・・・−0.05 parts by weight Sulfosalicylic acid・・−・−・−・−・−・・・・・・・・・・・・・・
...-50 parts by weight Sorbitan monolaurate (manufactured by Kao, Rheodor 5P-LIO)
・4 parts by weight cetyltrimethylammonium bromide・−・−・−・−・・−・・−・・−・・・−・・−
... 2 parts by weight Cellulose fine powder (Asahi Kasei, Avicel TG-D) - 155 parts by weight Methyl vinyl ether/maleic anhydride copolymer (GAF Co., Ltd.) amyl alcohol ester of Gantorezotsu AN-169)...
...10 parts by weight methyl cellosolve...
・−・・−・・・−・−・−・−160 parts by weight Methyl cellosolve acetate ・・・・・・65 parts by weight Results of using the same test liquid as used in Example 1 The coloration was as follows.

■－・・－・・・－・No discoloration.

■・・〜・−・−・Pale pink coloration.

■・・・・・・；−・Pink coloration.

■−・・−・・−・No discoloration.

4.4° -Bis-diazonium diphenylmethane tetrafluoroborate・・・・・・−・−・−・−0.2 parts by weight Metaphosphoric acid・・−・−−11 −−−−−−・・−・・・・・
・・・−・・・・・・・−・−−−−−−−40 parts by weight Sodium dodecyl sulfate−−−−−−−・−・−・−
......6 parts by weight Cellulose fine powder (Asahi Kasei, Avicel TG-D) 160 parts by weight Methyl vinyl ether/ Maleic anhydride copolymer (GAF
Amyl alcohol ester of GANTRESOTSU AN-169) manufactured by MITSUMI CO., LTD.
−−・−・−10 parts by weight methyl cellosolve−・・−・−
・−・−−−−11111・・・・・160 parts by weight Methyl cellosolve acetate−・・・・・・80 parts by weight The same liquid to be tested as used in Example 1 was used. The resulting coloration was as follows.

■−・−・−・・No discoloration.

■−・−・−・−・・Pale pink coloration.

■－・・・・・・－Pink coloration.

■－・－・・・－・No discoloration.

1 Jel- A test piece was obtained in the same manner as in Example 3, except that a test composition having the following composition was finely dispersed using a homomixer and dried after printing at 40°C for 70 minutes. .

Issei U Rule" Ll p-Dimethylaminobenzaldehyde...-----1...-----0.8 parts by weight Sulfosalicylic acid--...・・・・・・
-・・・-・-・15 parts by weight Sorbitan monolaurate (manufactured by Kao, Rheodol 5P-LIO) −・−・・−・・1.3 parts by weight Cellulose fine powder (manufactured by Asahi Kasei, Avicel 5F) -・・・・−・−・・−・−・・40 parts by weight Methyl vinyl ether/maleic anhydride copolymer (manufactured by GAF,
Amyl alcohol ester of Gantorezotsu AN-169)
・−・・−・−・10 parts by weight amyl alcohol−・・・・
・−・・−・・・−・−・−・・−・−・−・
-351!16B Among the test liquids similar to those used in Example 1, when ■■ was used, it was immersed in the test liquid and taken out, and 60 seconds after leaving it still, ■-...・Colored yellowish brown.

■－・・・－・・・・・・Colored yellowish brown.

(■1 mg7d l of urobilinogen-containing ring) (■
5 ■g/dl of urobilinogen-containing ring), but the difference between the coloring density of ■ and the coloring density of ■ was only slightly recognized, and in the case of Examples 1 to 4. It was smaller than the difference between the color density of ■ and the color density of ■.

1. A test composition having the following composition was prepared, and a test piece was obtained in the same manner as in Example 1.

4,4-bis-diazonium tetrafluoroborate 0.2 parts by weight meclic acid
・−−−１−−・−・・・・・−−−−−１・・・・−
・−−−1−−−−−・−・・・・・・・−6 parts by weight Sodium dodecyl sulfate−−−・・−・・・・・・0
．． 6 parts by weight methyl alcohol--・・・・-・-・・
−・・−・・−・・−・−・tsx parts water・−・−・−
・・−・・−・−・−・−・−・・−・・−・−・・
・・−・−・−１−−−−−・−・・・−・−−−−−
-85 parts by weight The obtained test piece was immersed in 5 tagged 1 urobilinogen-containing ring (same as ``■'' in each of the above examples), taken out, and allowed to stand for 20 seconds, giving it a reddish-purple color.

Separately, 21g of the untested test piece obtained above was
/d linogen immersed in ffi bilirubin-containing ring (lower concentration of ■ in each of the above examples), 2
After being immersed in urine containing bilirubin at mg/d 1, it was immediately taken out and allowed to stand for 40 seconds, resulting in a dark greenish-brown color, confirming that bilirubin had a large effect.

Claims (2)

[Claims] (1) General formula▲There are mathematical formulas, chemical formulas, tables, etc.▼・・・・・・・・・
・[1] ▲There are mathematical formulas, chemical formulas, tables, etc.▼・・・・・・・・・
・[2] ▲There are mathematical formulas, chemical formulas, tables, etc.▼・・・・・・・・・
・[3] ▲There are mathematical formulas, chemical formulas, tables, etc.▼・・・・・・・・・
・[4] (In the formula, (X)^- and (Y)^- represent the same or different stabilizing anions, and Ph represents a substituted or unsubstituted phenylene group.) A test composition for detecting urobilinogen, characterized in that it contains at least one. (2) General formula▲There are mathematical formulas, chemical formulas, tables, etc.▼・・・・・・・・・
・[1] ▲There are mathematical formulas, chemical formulas, tables, etc.▼・・・・・・・・・
・[2] ▲There are mathematical formulas, chemical formulas, tables, etc.▼・・・・・・・・・
・[3] ▲There are mathematical formulas, chemical formulas, tables, etc.▼・・・・・・・・・
・[4] (In the formula, (X)^- and (Y)^- represent the same or different stabilizing anions, and Ph represents a substituted or unsubstituted phenylene group.) A test piece having, as a part of a support, a test composition for detecting urobilinogen, which contains at least one composition.