JPH0315399A - Composition for testing humor ingredient - Google Patents

Abstract

PURPOSE:To readily enable obtaining of a composition, having a constant quality and excellent in preservation performance with high test accuracy by dispersing and kneading a physiologically active substance, such as an enzyme with a binder and water absorbing fine particles in a nonaqueous solvent. CONSTITUTION:A physiologically active substance, produced by an enzyme, etc., reactive with pathological ingredients, such as cholesterol, contained in humor and causing a change in color tone to an indicator, a buffer agent, the indicator, etc., as necessary, added thereto are pulverized into particles having <=about 25mu particle diameter. The resultant pulverized substance and water absorbing fine particles, such as silica-based clay or cellulose, are dispersed and kneaded with a nonaqueous solvent, such as alcohols, in which a binder, such as isobutylene-maleic anhydride copolymer, is predissolved utilizing a high-speed stirrer, etc., to afford a composition for testing humor ingredients. The resultant composition is applied to a substrate for specimens by utilizing a well-known printing method, such as silk screen printing. Thereby, the objective product of good quality for the specimens of the humor ingredients can be produced on an industrial scale.

Description

[Detailed description of the invention] [Industrial application field]

The present invention is applicable to blood, lymph fluid. This article relates to a composition for testing body fluid components, which is used as a specimen for testing pathological components contained in body fluids such as urine.

[Conventional technology]

Testing for pathological components contained in body fluids such as blood, lymph, and urine is essential for disease diagnosis and health management, and tests using various test specimens are essential. It is done l 2 . Conventional test specimens for testing the above-mentioned pathological components utilize changes in color due to chemical reactions between target components in body fluids and reagents, but in recent years, enzymes, etc. Testing methods that use physiologically active substances are becoming more popular because they do not require the use of strong acids or alkalis, and can test for pathological components in body fluids under relatively mild conditions. It has become. For example,
No. 1878 discloses that a liquid enzyme, a substance having a peroxidation reaction function, a buffer, orthotolidine dihydrochloride, copolymers such as polyvinylpyrrolidone, and a surfactant are added to a hygroscopic material. A test specimen for glucose detection obtained by impregnating a liquid mixture consisting of The conventional test specimen is made of water and water so that the color of the test reagent layer changes depending on the content of the test target substance in the test liquid.
After impregnating a hygroscopic material such as a filter paper in multiple stages with a solution in which enzymes, indicators, etc. are dissolved in an alcohol-based mixed solution, the resulting impregnated paper is cut into appropriate sizes. It is manufactured by attaching it to the substrate for the test object.

[Problem to be solved by the invention]

By the way, in the conventional body fluid test specimens using enzymes, the enzyme dissolved in the water/alcohol mixed solution during the process of obtaining the test specimens is unstable and deteriorates rapidly over time. To do this, the hygroscopic material must be impregnated in multiple stages immediately after the impregnation solution is prepared, and the manufacturing process is complicated. has drawbacks such as requiring special skill. In contrast, the present invention is a body fluid component testing composition used when obtaining a body fluid component test body, which exhibits extremely excellent preservation performance, has high testing accuracy, and has consistent quality. To provide a composition for testing body fluid components, which allows a test sample to be obtained extremely easily.

[Means for Solving the Problems] The composition for body fluid component testing of the present invention is applied to a base material for a body fluid component test body when obtaining a body fluid component test body, and is capable of containing physiologically active substances such as enzymes. It consists of a composition in which a binder and water-absorbing fine particles are dispersed and kneaded in a non-aqueous solvent. Each component used in the body fluid component testing composition of the present invention having the above structure will be explained below. Pathological components contained in body fluids, such as glucose, cholesterol, urea. Neutral fat, phospholipid. Ammonia, lactic acid, pyruvic acid, sialic acid, GPT, G
OT, cholinesterase, leucine aminobebutitase, amylase, LCAT. A physiologically active substance such as an enzyme that reacts with taleatin phosphokinase etc. is used and causes a color change to the indicator, or a substance produced by the reaction reacts with another physiologically active substance such as an enzyme. However, those that cause a change in color tone relative to the indicator are used. [Conclusion] The L-shaving bonding agent is used to fix the body fluid component testing composition of the present invention to a base material for a body fluid component test body, and to cover the composition of the present invention that is fixed to the base material for the body fluid component test body. It prevents elution into the test fluid, and in particular, the pathological components contained in the aforementioned body fluids and other components in the composition of the present invention, such as physiologically active substances such as enzymes, etc. It goes without saying that materials should be used that do not affect the body's health or interfere with testing of body fluid components. It is desirable to check whether the binder satisfies the above-mentioned conditions each time before using the binder, but in the experiments conducted by the present inventors, for example, polyacrylic ester, polyphenylacecur, Polymethacrylate. Synthetic polymers such as polyacrylamide, polyurethane, polyvinyl chloride, polystyrene, maleic acid copolymers, polyvinyl acetate, polyvinyl alcohol, polyvinyl pyrrolidone, and methylcellulose. Cellulose derivatives such as ethyl cellulose and probil cellulose, semi-synthetic polymers such as starch ether, starch, and casein. Natural polymers such as polysaccharides can be used. In particular, when a maleic acid copolymer such as inbutylene-maleic anhydride copolymer or styrene-maleic acid copolymer is used as a binder, the binder must have good shape retention performance and good liquid stability. This is advantageous because it provides good permeability. The water-absorbing fine particles increase the water-absorbing and water-retaining properties of the test area layer obtained by applying the body fluid component test composition of the present invention to a substrate for a test body, and can absorb enzymes, etc. It has the effect of smoothly promoting the reaction between the physiologically active substance, the indicator, and the pathological component that is the target component of the test.For example, silica clay. Inorganic or organic fine powders such as calcium carbonate, aluminum silicate, glass, and cellulose are used. The composition for body fluid component testing of the present invention is composed of a mixture of a non-aqueous solvent containing a physiologically active substance such as an enzyme, a binder, and water-absorbing fine particles as essential components. In addition, for example, anionic, thionic, or nonionic surfactants may be added as necessary to improve the water wettability of the buffering agent, test area layer, uniformity of color development, etc. It will be done. The nonaqueous solvent used for cross-mixing the components is, for example, an organic solvent such as alcohols, ketones, aromatics, esters, or hydrocarbons, and a solvent that can dissolve the binder is selected. Used. Furthermore, if the non-aqueous solvent used contains water,
Since the activity of physiologically active substances such as enzymes in the composition is rapidly lost, it is preferable to use an organic solvent that has been dehydrated in advance. Among non-aqueous solvents, methanol is
Since it tends to reduce the activity of physiologically active substances, albeit slowly, it is preferable to avoid it if possible. Furthermore, an indicator for performing a color reaction in a body fluid component test body obtained using the composition for body fluid component test of the present invention is contained together with essential components in the composition for body fluid component test. Good. This indicator uses a physiologically active substance such as the enzyme mentioned above that acts with a pathological component and changes its color tone. Physiologically active substances such as enzymes and indicators are selected and used as appropriate from among the many combinations known for detecting pathological components. For example, for glucose detection, enzymes... ...Glucose oxidase indicator...Mechanism in which the penzidine system is used, glucose oxidase oxidizes glucose, and the hydrogen peroxide produced at the same time changes the color of the benzidine system indicator in the presence of the system oxidase. is used. In addition, for uric acid detection, a mechanism is used in which uricase is activated and the generated hydrogen peroxide is colored in a reaction system equipped with a chromogen. Furthermore, for cholesterol detection, a mechanism is used in which cholesterol oxidase is activated and the generated hydrogen peroxide is colored in a reaction system equipped with a chromogen. The composition for body fluid component testing of the present invention includes pulverized physiologically active substances such as freeze-dried enzymes, buffers and indicators added as necessary, and
After pulverizing into particles of approximately 25 μm or less, these pulverized particles and water-absorbing fine particles are dispersed in a non-aqueous solvent in which a binder has been dissolved in advance. It is easily obtained by kneading. In addition, this section. The operation of crosstalk is, for example, high-speed stirring I.
I. Sand mill, ball mill. This is done using a homogenizer, three rolls, ultrasonic disperser, etc. The base material for the test body to which the composition for testing body fluid components of the present invention having the above-mentioned structure is applied includes the pathological components contained in the above-mentioned body fluids and the composition for testing body fluid components of the present invention. Any material may be used as long as it does not affect or interfere with body fluid testing, such as paper, synthetic paper, and various plastic films.
Complexes of two or more of these types are used. In order to apply the body fluid component testing composition of the present invention to a substrate for a test body, it is convenient to use a known printing method. In particular, printing methods that allow printing with a relatively large coating amount and a constant coating amount, such as silk screen printing, intaglio printing, gravure printing, etc., are suitable. The amount of the composition for body fluid component testing is usually 2 to 2.
30g (solid component) of approximately 7m''.

[For use]

The composition for body fluid component testing of the present invention is applied to a base material for a body fluid component test body when obtaining a body fluid component test body, and includes a physiologically active substance such as an enzyme, a binder, and water-absorbing fine particles. It consists of a composition in which the essential ingredients are dispersed and kneaded in a non-aqueous solvent. The composition for body fluid component testing of the present invention having the above-mentioned configuration is stable because the physiologically active substances such as enzymes in the composition are difficult to deteriorate, and therefore the composition can be stored and used for body fluid component testing. Since the composition is easy to handle during production, a body fluid component test sample of good quality can be easily obtained. In the composition for testing body fluid components of the present invention, that is,
It is not clear why physiologically active substances such as enzymes are difficult to alter and remain stable over long periods of time in body fluid component testing compositions that are contaminated with non-aqueous solvents. Water-soluble physiologically active substances exist in a dispersed state without being dissolved in non-aqueous solvents, so they are less susceptible to denaturation at the higher-order structures and active sites of the proteins that make up the physiologically active substances. This is presumed to be due to the fact that it is difficult for the solvent to penetrate into the interior of the physiologically active substance even if the area near the interface of the substance is denatured, and the activity of most of the physiologically active substances is not impaired. ．． This is also supported by the fact that the activity of physiologically active substances such as enzymes rapidly decreases in a mixed solvent of non-aqueous solvent and water.

【Example】

Hereinafter, the specific structure of the composition for testing body fluid components of the present invention will be explained based on Examples, and the effects of the composition for testing body fluid components and the effects of using the composition for testing body fluid components will be explained below. We will explain the effects of the body fluid test specimens obtained using this method. Example 1 A test composition for glucose detection [al], which is an example of the present invention, was prepared by finely dispersing a composition having the following composition using a homomixer. Glucose oxidase: 0.5 part by weight {Toyobo Co., Ltd., Grade II) Peroxidase: 0.1 part by weight {Toyobo Co., Ltd.
(Strain 1, Grade III}0-tolidine...
・・・・・・・・・・・・・・・0.5 parts by weight Polyoxytylene sorbicun monooleate・・・1.0 parts by weight Isobutylene/
Maleic anhydride copolymer: 3.0 parts by weight {kurarain butylene chemical, isobane #60} n-butanol: 50. 0 parts by weight Kaolin 45.0 parts by weight [Experiment 1] Using the obtained test composition [a] above, screen-screening was performed on a base plate for a test specimen made of a white polystyrene film having a thickness of 300 μm. Due to the printing method, one side is 61II
After printing the I+ square pattern, it was dried at 80°C for 1 hour. The mesh of the screen plate used in the above printing was 100, and the total thickness of the resist and screen gauze was approximately 100μ. Next, the white polystyrene film was
It was cut into a rectangular piece of I O Om+a with the square pattern printed portion located at one end of the resulting rectangular piece to obtain a body fluid component test body. The obtained body fluid component test sample was quickly immersed in urine containing a known glucose concentration, and the color tone was observed after 30 seconds. The results are shown in Table 1. Note that the color tone indications in Table 1 are based on JIS Z872.
6 according to the standard color standard according to l. 1st
From the results shown in Table 1, it is possible to measure urine containing a glucose concentration of 0.05% by using the test sample obtained with the composition for body fluid component testing of the example product. It is clear that Comparative Example 1 An impregnating solution [b] was prepared using a composition having the following composition. Glucose oxidase...0.6 parts by weight {Toyobo Co., Ltd. {Strain 1, Grade II} Peroxidase...{Toyobo Co., Ltd., 0.15 parts by weight Grade I■} 0-Toridine dihydrochloride・・・・・・ 0.5 parts by weight Citric acid monosodium citrate buffer (pi{=5.51・・・・・・・・・・・・・・・
80 parts by weight Gelatin aqueous solution (5% by weight) 82.5 parts by weight Distilled water 20 parts by weight Ethyl alcohol 23.7 parts by weight [Experiment 2] Obtained impregnating solution [bl is filter paper (Toyoro), and after cutting the impregnated paper into a square with one side of 6n+m,
! A body fluid component test sample was obtained by attaching a double-sided adhesive transfer tape (3M) to one end of a white polystyrene film in the form of a strip of IIIX 1 0 011111. Comparative Example 2 An ink composition having the following composition [cl was adjusted. Glucose oxidase...0.4 parts by weight {Toyobo Co., Ltd. 1, Grade II 1 peroxidase...
・・・・・・・・・0.・l parts by weight {Toyobo Co., Ltd. 1
, Grade III}0-tolidine...
・・・・・・・・・0.5 parts by weight Styrene/maleic acid copolymer resin solution {Seiko Kagaku Kogyo Co., Ltd. 1, X-1
202LLl...30.0 parts by weight PU buffer solution (pH=7.01...20
．． 0 parts by weight Ethyl alcohol ・・・・・・・・・l5
, 8 parts by weight kaolin ・・・・・・・・・・・・・・・
- 15.0 parts by weight [Experiment 3] After printing a pattern of 11 squares on each side with the obtained ink composition [cl] on a base plate for a test object made of a white polystyrene film with a thickness of 300 μm, It was dried at 45°C for 2 hours. The mesh of the screen plate used in the above printing was 100, and the total thickness of the resist and screen gauze was approximately 100μ. Next, the white polystyrene film was shaped into a strip of approximately 6+++ mX I O Osm.
The resulting strip was cut so that the square pattern printed portion was positioned at one end to obtain a body fluid component test piece. [Experiment 4] The above [Experiment 1],

Each body fluid component test sample obtained in [Experiment 2] and [Experiment 3] was briefly immersed in urine containing a known glucose concentration, and the color tone was observed after the elapse of a preset determination time. The results are shown in Table 2. In addition, when determining the color tone, the elapsed time at which the change in color tone due to glucose for each test specimen was most noticeable was tested in advance, and that time was selected as the elapsed time required for the determination. Table 2 From Table 2 above, it is clear that the test results from the body fluid component test specimen obtained using the body fluid component test composition of Example 1 have excellent quantitative properties. Furthermore, the stability of the color tone after color development was also good. Example 2 The composition for a body fluid test body obtained in Example 1 [al] was left at 40°C for 1 to 7 days to produce a composition for a body fluid test body, which is yet another example product of the present invention. I got something. [Experiment 5] Using a base material for a test body made of a white polystyrene film with a thickness of 300 μm and the composition for a body fluid test body obtained above, the above-mentioned procedure was carried out.

A body fluid component test sample was obtained using the same procedure as the corresponding step in [Experiment 1]. Comparative Example 3 After adjusting the impregnating solution [bl] obtained in Comparative Example 1, it was left to stand at 40°C for 1 to 7 days to obtain an impregnating solution for comparison.

[Experiment 6] The impregnation solution obtained in Comparative Example 3 and filter paper (Toyo Roshi,
No. A body fluid component test specimen was obtained using the same procedure as the corresponding step in [Experiment 2] described above. Comparative Example 4 The ink composition obtained in Comparative Example 2 was left at 40° C. for 1 to 7 days after adjusting the Cl to obtain a composition for body fluid component testing for comparison. [Experiment 7] Composition for testing body fluid components obtained in Comparative Example 4 and thickness 300
Using μ white polystyrene film, the above-mentioned

A body fluid component test sample was obtained using the same procedure as the corresponding step in [Experiment 3]. [Experiment 8] Each of the body fluid component test specimens obtained in [Experiment 5], [Experiment 6], and [Experiment 7] described above was quickly immersed in a 0.5% glucose solution. ] The coloring density after the corresponding judgment time elapsed was measured with a reflection densitometer and compared with the coloring density obtained in the previous [Experiment 4].

Table 3 shows the relative concentration (%) of each body fluid component test sample when the coloring density obtained in [Experiment 4] is taken as 100. Table 3: Umbrella: It is impossible to prepare a test specimen due to the ink being made into a genol. From the results shown in Table 3, the body fluid component test was performed using the composition for body fluid component test according to Example 2. The quality of the body remains unchanged even if the test composition is not immediately prepared, but the body fluid component test specimens obtained using the impregnating solutions and ink compositions according to Comparative Examples 3 and 4 do not change in quality. Compared to specimens manufactured by applying an impregnating solution or ink composition to a substrate for specimens immediately after preparation, the quality of the obtained body fluid component specimens deteriorates as the storage time increases. It is clear that this decreases.

【Effect of the invention】

As is clear from the experiments based on the above-mentioned Examples and Comparative Examples, the composition for body fluid component test body of the present invention has extremely good storage stability, and the test area layer formed by the composition has excellent storage stability. The color reaction is clear depending on the content in the fluid being tested, making it a body fluid component test material that can easily provide detailed test results. Furthermore, since the composition for body fluid component testing of the present invention can be applied to a substrate for a test body by ordinary printing means, it is possible to manufacture a body fluid component test body when using the composition. is extremely simple, and high-quality products can be mass-produced on an industrial scale.

Claims (1)

[Claims] 1. A composition for testing body fluid components, comprising a composition in which a physiologically active substance, a binder, and water-absorbing fine particles are dispersed and kneaded in a non-aqueous solvent.