JPS6010261B2 - Composition for detecting occult blood - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a significantly improved method useful for the qualitative and quantitative determination of blood in body fluids such as urine, vomit, gastrointestinal contents, cerebrospinal fluid, feces, and waste materials. The present invention relates to a composition for detection.

Detection of occult blood in body fluids and bodily excreta and knowing its amount provides valuable information for diagnosing diseases and observing the progress of prognosis, and is currently an important test indispensable in clinical medicine. ing.

That is, in diseases that involve erosion of the mucous membranes, such as thin water and cancer, blood is found in the stomach contents and vomit, and latent blood is increasingly seen in feces. From the viewpoint of clinical medicine, a submersible dish detection method is required that can reliably detect even minute amounts of occult blood that cannot be detected by microscopic identification of blood in body fluids or body excreta. In addition, blood components or blood molecules may be present in the urine as a result of typhus fever, scurvy, purpura, cerebral hemorrhage, hives, hives, burns over large parts of the body, or the effects of various engorged toxins. It is even more important to be able to detect occult blood in urine since it is excreted. There are various methods for detecting occult blood in body fluids or biological wastes, but the main method currently in use is to detect the oxidation state of blood components hemoglobin, myoglobin, or decomposition products by alternating with an oxygen source. If the receptor is a dye precursor, it is colorless until it becomes oxidized, but it develops color in the oxidized state. The presence of hemoglobin, a blood component, or its decomposition products can be detected from the color tone that appears, and the presence or absence of bleeding, as well as its amount, can be determined.

The speed of color change and the intensity or density of the color provide a means of quantifying the amount of blood present. However, these test methods have drawbacks such as requiring measuring instruments or devices and being complicated to operate.Although diagnostic test strips were considered that would be easy for beginners to perform measurements, they lacked sufficient sensitivity. However, there is a need for even better test compositions because of problems such as poor performance and insufficient stability of the test paper.

The present inventors have conducted extensive research on methods for measuring blood that have such drawbacks, and have found that acridine has been added to various detection compositions in methods for measuring occult blood that utilize reagent compositions consisting of indicators, buffers, and peroxides. , 2-methoxyacridine, phenanthridine, 506-benzoquinoline, 7.8
- By adding benzoquinoline or their organic acid salts and key salts such as hydrochloric acid or sulfuric acid, unexpectedly,
He completed the present invention by discovering that the speed of melancholy is accelerated, the sensitivity is increased, and the sensitivity is maintained for a long period of time.

As mentioned above, the hemolysis test was performed using acridine, 2-methoxyacridine, phenanthridine, 5,6-benzoquinoline,
This can be improved by adding 7,8-benzoquinoline or an organic acid salt thereof and a key salt such as hydrochloric acid or sulfuric acid.

For example, cumene hydroperoxide o-tolidine, p
Table 1 shows the color development of a test piece obtained by impregnating and drying a paper with a solution prepared by adding several different synergists of the present invention to a submersible dish measurement composition consisting of an H5 citric acid buffer. become.

Also shown in the same table are comparative examples in which quinoliso, 6-methoxyquinoline, quinine, and pyridine were used as synergists instead of the compounds of the present invention. However, 1' blood
After immersing the test piece in a solution diluted to 250,000, 1'100,000, 1'300,000, 1/600,000, take it out immediately.3 No star color detected after sanding (1) Slight star color. It was measured in four stages: (10), normal coloration (20), and deep coloration (30). Table 1 *: Slightly darker than that of female and black.

Note) For pyridine, use an undried test piece. (This is because pyridine evaporates when dried.) Data regarding the stability of the composition is also shown in Table 2. However, 1 blood
After the test piece was immersed in the diluted solution, it was immediately taken out and the coloration after 3 quarts was measured. The present invention is an organic hydroperoxide, a substance capable of maintaining the pH of the substance to be tested in the range of 4 to 7, and a substance that oxidizes with a color change due to the presence of a blood complementing molecular group present in blood. The indicator used is one or more selected from the group consisting of acridine, 2-methoxyacridine, phenanthridine, 5,6-benzoquinoline, 718-penzoquinoline, or their organic acid salts and key salts such as hydrochloric acid or sulfuric acid. By coexisting the compound, it has excellent sensitivity and storage stability.
Although it is possible to provide a composition for detecting erythremia in body fluids and body excreta, in carrying out the present invention, organic hydroperoxides such as cumene hydro/gu oxide, diisopropylbenzene, etc. Hydro/Goo Oxide, /f Ramentane Hydro/
Examples include goo oxide, 215-dimethylhexane-2,5-dihydroperoxide, and the like.

In general, organic hydroperoxides are unstable and easily decompose especially when they come into contact with various other substances, and they also alter the quality of the pond material, so it is effective to isolate them as microcapsules using encapsulating materials as a means of isolation. The encapsulating material used can be one or a mixture of two or more of various proteins or polysaccharide materials, such as gelatin, arginisotope, carrageenan, gum arabic, casein, albumin, etc. The encapsulating material and encapsulation method It is not limited to. Useful buffers include, for example, tartaric acid, phosphoric acid, phthalic acid, citric acid, acetate buffers, etc., and the preferred range for buffering in the composition is from pH 4 to pH 7. Various indicators can be used as indicators that are oxidized with a color change in the presence of hemoglobin complement molecules. Examples include aniline and phenol derivatives, such as o-toluidine, p-toluidine, o-phenylenediamine, N.N'-dimethyl-p-phenylenediamine, N.N'-diethyl-p-phenylenediamine, benzidine, p-anisidine, dianisidine,
Examples include o-tolidine, o-cresol, m-cresol, p-cresol, Q-naphthol, 8-naphthol, catechol, guaiacol, pyrogallol, and the like. The novel composition of the present invention can be used in various forms for diagnostic purposes, and impregnating a water-absorbing material such as paper or fiber with the composition in the form of a disk or stick is the most effective means in practice. becomes.

The present invention provides a reagent composition for detecting port blood with high sensitivity and excellent stability, and provides a submersible plate detection method that can be easily and reliably performed without requiring any skill. There are great things that can contribute to therapeutic medicine. Next, examples of the present invention will be shown and explained in detail.

Example 1 30% aqueous solution of gum arabic 600 g
Heat to 60°C, add 40°C of cumene hydroperoxide, add 300ml of pH 5 citric acid solution containing 5% gelatin, and keep at 60°C.

Cool this with water and pH 5 citric acid softening solution (0.8 core 30
After soaking the monkey paper in test solution 1 prepared by adding 0' and mixing thoroughly, put it in a dryer and dry it at 60-8000 °C.
A test solution prepared by dissolving 3 nights of -tolysine and 1 night of acridine in 1 night of chloroform is immersed in a funnel, and then dried. Example 2 According to Example 1, test solution 1 was prepared and impregnated with dried o-tolysine for 3 nights and 7,8-benzoquinoline for 1 night.

After impregnating it with test solution 0 prepared by dissolving it in form 1, it is dried. Example 3 According to Example 1, test solution 1 was prepared, impregnated with a test solution prepared by dissolving 1 solution of otolysine and 5,6-penzoquinoline in 1 solution of chloroform, and then dried. do.

Example 4 According to Example 1, a test solution 1 is prepared, and after impregnating and drying, it is impregnated with a test solution 0 prepared by dissolving 10 minutes of o-tolysine and 5 hours of phenanthridine in chloroform, and then drying.

Example 5 Example 1 is followed.

However, diisopropylbenzene hydroperoxide is used instead of cumene hydroperoxide.
Example 6 Example 1 is followed.

However, paramenthane hydroperoxide is used instead of cumene hydroperoxide. Example 7 Example 1 is followed.

However, instead of the citric acid buffer solution (0.9M) for plaque 5,
of acetate buffer (0.8M) is used. Example 8 Example 1 is followed.

Claims (1)

[Claims] 1. Acridine in a composition for detecting occult blood consisting of an organic hydroperoxide, a buffer that maintains the pH in the range of 4 to 7, and an indicator that is oxidized with a color change in the presence of a blood complement group present in the blood. , 2-methoxyacridine, phenanthridine, 5,6-benzoquinoline, 7.
8-Benzoquinoline or one or more compounds selected from the group consisting of organic acid salts thereof and mineral acid salts such as hydrochloric acid or sulfuric acid, for detecting occult blood in body fluids and body excreta. Composition.