KR102480846B1 - Solid beads for diagnosing proteinuria in cat urine and proteinuria diagnostic kit containing the same - Google Patents

Abstract

The present invention relates to a solid bead for diagnosing proteinuria in cat urine and a kit for diagnosing proteinuria using the same, and in detail, includes a tetrabromophenol blue indicator; and an acid solution having a pH of 2 to 3; and a water-soluble polymer including cellulose or alginate or a mixture thereof; placed on alkaline sand such as bentonite, and when the cat urinated, the solid bead Because it is simple and easy to check the presence of proteinuria through color change, it can be widely used as a cat proteinuria diagnosis kit.

Description

Solid beads for diagnosing proteinuria in cat urine and proteinuria diagnostic kit containing the same {Solid beads for diagnosing proteinuria in cat urine and proteinuria diagnostic kit containing the same}

The present invention relates to a solid bead for diagnosing proteinuria in cat urine and a kit for diagnosing proteinuria using the same, and in detail, includes a tetrabromophenol blue indicator; and an acid solution having a pH of 2 to 3; and a water-soluble polymer including cellulose or alginate or a mixture thereof; the solid bead prepared by mixing the solid bead is placed on the alkaline coagulated sand and the color of the solid bead when the cat urinates Because it is simple and easy to check the presence of proteinuria through changes, it can be widely used as a cat proteinuria diagnostic kit.

In modern society, the perception of pets, including dogs and cats, has changed due to urbanization and the increase in single-person households, making them a culture that is not just an animal but a member of the family. In particular, the social atmosphere to find companion animals such as dogs and cats as a means to gain familiarity with nature and animals and to know nature through sympathy and improve the quality of life in the city center is expanding, starting from animal lovers and maniacs. Demand for companion animals is diversified and gradually increasing. Currently, companion animals are widely raised in ordinary homes, and there is a trend that goes beyond the concept of animals and takes their place as family members and life partners. In particular, cats are easier to raise indoors than dogs, take less time to raise, and have higher convenience for raising cats, so the number of raising objects is rapidly expanding from countries where companion animal culture has developed earlier, such as Japan and Europe. It is increasing.

Changes in perception of companion animals lead to an aging era of companion animals along with lifespan extension, and chronic and old-age diseases that have been found and treated in humans are becoming major diseases in companion animals. Among these, kidney disease is a major disease that is fatal to cats, as it is the number one cause of death in cats because it is not only a risk for most companion cats due to the lifestyle of cats, but also is related to diabetes, urinary stones, bacterial cystitis, etc., and is difficult to cure. being reported as Proteinuria is a representative item that is an indicator of kidney disease, and if the symptoms of proteinuria are left unattended, it is highly likely to cause serious damage to the kidneys and develop into chronic renal failure. to be.

Kidney disease is difficult to cure once it occurs, has a high probability of recurrence, and has the highest mortality rate within 6 months after diagnosis. Therefore, early detection has a significant impact on the cat's lifespan, so a quick examination is necessary. However, in the case of cats, it is very difficult for non-experts to test proteinuria at home due to regular inspection and early diagnosis due to difficulties in urine collection.

On the other hand, in the case of cats, unlike other companion animals, they have the characteristic of burying feces in sand when they defecate or urinate, so sand (aka cat sand) is essential for this, and without sand, smooth bowel movement is prevented. do. For this purpose, cat litter and toilet articles have been developed and used in various ways, and in the case of cat litter, depending on the method of use, it is largely classified into absorption type sand and coagulation type sand.

The absorbent sand is a product that has a function of absorbing cat feces, especially urine, and uses silica (SiO ₂ ㆍ 2H ₂ O), which is used as a desiccant, or paper sand or grains such as corn and beans. This is used, and the sand obtained by crushing bentonite is used as the coagulation type sand. In particular, coagulation type sand using bentonite is a product with excellent excrement treatment by aggregating excrement with bentonite sand, and is most widely used in households raising cats.

However, although the utilization and consumption of bentonite sand is superior to other absorbent sands, silica gel sand has been the only indicator used to test proteinuria in cat litter due to the special properties of bentonite.

Under this background, the present inventors tried to solve the error of existing indicators for detecting proteinuria in silica gel sand and to develop a diagnostic composition capable of simply and accurately diagnosing proteinuria in cat urine in alkaline coagulated sand such as bentonite. .

Republic of Korea Patent Publication No. 10-2017-0002318 (2017.01.06. Publication)

An object of the present invention is to provide a diagnostic composition capable of simply and accurately diagnosing proteinuria in cat urine in bentonite coagulated sand and a method for diagnosing proteinuria using the same.

The present invention relates to (a) 100 parts by weight of a water-soluble polymer containing cellulose or alginate or a mixture thereof, (b) 0.04 to 0.1 part by weight of tetrabromophenol blue indicator; and (c) 1.2 to 5 parts by weight of an acid solution having a pH of 2 to 3.5.

In addition, the present invention (i) 0.04 to 0.1 parts by weight of a tetrabromophenol blue indicator based on 100 parts by weight of a water-soluble polymer containing cellulose or alginate or a mixture thereof; and mixing and kneading 1.2 to 5 parts by weight of an acid solution having a pH of 2 to 3.5; (ii) putting the dough into a molding machine and processing it into a bead shape; and (iii) drying the processed product in the form of a bead.

In addition, the present invention the solid bead; and a kit for diagnosing feline proteinuria comprising bentonite coagulated sand.

In addition, the present invention comprises mixing the solid beads with bentonite solidified sand; And it provides a method for diagnosing proteinuria, including the step of confirming the color change when the cat's urine comes into contact with the solid beads.

According to the present invention, it is possible to provide a method for detecting proteinuria using solid beads that can simply and accurately diagnose proteinuria in cat urine in bentonite coagulated sand, so that kidney disease can be diagnosed at an early stage without separate cat urine collection. It is expected to contribute to the extension of the life span of companion cats, as well as to lower the burden of medical expenses for cat guardians and improve the quality of life.

1 is a schematic diagram showing that the color changes to blue in response to tetrabromophenol blue indicator and proteinuria.
Figure 2 is a schematic diagram showing the color change when tetrabromophenol blue indicator is sprayed on the surface of bentonite sand and reacted with proteinuria or distilled water.
Figure 3 shows the appearance of a solid bead according to the present invention.
4 is a result showing that the existing indicator used in silica gel sand is raised on bentonite sand, and distilled water and proteinuria are sprayed, and then the distilled water turns green.
5 is a result of confirming the color change after mixing bentonite and the existing indicator used in silica gel sand at a ratio of 1: 1 and sprinkling distilled water and proteinuria, and is a result showing that the color change could not be confirmed.
6 shows the results of diagnosis of proteinuria in bentonite sand using solid beads prepared using benzoic acid, citric acid, and tartaric acid.
FIG. 7 is a result of numerically calculating a color value difference based on the result of FIG. 6 .

Hereinafter, the present invention will be described in more detail.

The present inventors tried to devise a method for diagnosing proteinuria in urine in a convenient and easy way in bentonite sand used for raising cats. In the case of solid beads containing an acid solution along with tetrabromophenol blue indicator for the accuracy of diagnosis of proteinuria, an error in which the color change of the indicator used in existing silica gel sand occurs in solutions other than protein due to the strong alkali of bentonite sand It was confirmed that the accuracy of diagnosis can be improved by improving the bar, and the present invention has been completed.

In addition, the present invention (a) based on 100 parts by weight of a water-soluble polymer containing cellulose or alginate or a mixture thereof, (b) 0.04 to 0.1 parts by weight of tetrabromophenol blue indicator; and (c) 1.2 to 5 parts by weight of an acid solution having a pH of 2 to 3.5.

Here, the absorbent sand has a function of absorbing cat feces, especially urine, and uses silica (SiO ₂ ㆍ2H ₂ O), which is used as a desiccant, or paper sand or grains such as corn and beans. Absorption-type sand using is representative, and in the present invention, the solidification-type sand is preferably bentonite. In particular, coagulation type sand using bentonite is a product with excellent coagulability and is the most popular and used trend.

The water-soluble polymer is a formulation for preparing a solid bead for diagnosing proteinuria in cat urine in alkaline coagulated sand, and may be selected from cellulose or alginate or a mixture thereof. It is preferable to use microcrystalline cellulose as the cellulose. Cellulose, such as microcrystalline cellulose, is a material obtained from photosynthesis of plants, and can be used by hydrolyzing lignocellulosic cellulose, regardless of the presence or absence of lignin. . In addition, the alginate is a general term for those obtained from photosynthesis of marine organisms, and may be prepared and used by hydrolysis regardless of the type of organism, but is not limited thereto. Cellulose or alginate is preferably used because it has an advantage in that it can be molded and processed in the process of dispersing or dissolving it in water and drying it in an aqueous solution state, and more preferably microcrystalline cellulose and alginate It is used by mixing in a weight ratio of 15:1 to 30:1. This helps in forming solid beads due to the property of alginate to become hard after drying.

Furthermore, the water-soluble polymer of the present invention is carboxymethyl cellulose (CMC), methyl cellulose (MC), hydroxypropylmethyl cellulose (HPMC), hydroxypropyl methyl cellulose (HPMC), hydroxypropyl cellulose (HPC) and hydroxyethyl It may further include at least one selected from the group consisting of cellulose (HEC).

The tetrabromophenol blue indicator causes color development as the acidity of the pH indicator changes while meeting the charge of the peptide. The discoloration range is pH 3.0 to 4.6, and the acidic color develops yellow and the basic color develops blue.

The tetrabromophenol blue indicator is preferably used in an amount of 0.04 to 0.1 parts by weight based on 100 parts by weight of the water-soluble polymer. More preferably, it is 0.04 to 0.085 part by weight. In the case of less than 0.04 parts by weight, there is a limit that the color change may come out in the middle of yellow and blue instead of blue even in positive, and if it is more than 1 part by weight, there is a limit to change the color to positive at the protein concentration within the negative range within the above range good to use

The pH 2 to 3.5 acid solution plays the most important role in improving color change errors in solutions other than proteins due to strong alkalinity of bentonite sand, benzoic acid, citric acid, and tartar It is preferable to use at least one selected from the group consisting of acid (Tartaric acid). Among them, tartaric acid showed the most distinct color change.

The solid bead induces a color change by reacting with proteinuria in cat urine, and in particular, it may be prepared as a solid bead having a discoloration range at pH 3.0 to 4.6.

In addition, the solid bead composition may include at least one selected from the group consisting of cypress water, activated carbon, deodorant, phytoncide, and cypress oil, and these odor removal and antibacterial properties improve the product quality of the solid bead. can make it

In addition, the present invention (i) 0.01 to 0.03 parts by weight of tetrabromophenol blue indicator based on 100 parts by weight of a water-soluble polymer containing cellulose or alginate or a mixture thereof; and mixing and kneading 0.3 to 0.6 parts by weight of an acid solution having a pH of 2 to 3.5; and (iii) drying the processed product in the form of a bead.

In detail, the step (i) is a step of mixing the solid bead composition, and a solvent dissolving the tetrabromophenol blue indicator is additionally mixed to prepare the solid bead. The solvent may be at least one selected from the group consisting of ethanol, propanol, butanol, pentanol, hexanol, methanol, and dimethyl sulfoxide. The solvent is preferably used in an amount of 20 to 40 parts by weight based on 100 parts by weight of the water-soluble polymer. If it is less than 20 parts by weight, there is a limit that the tetrabromophenol blue indicator may not dissolve, and if it is more than 40 parts by weight, it does not play a role other than dissolving the tetrabromophenol blue indicator. Therefore, it is recommended to use it within the above range.

In addition, when mixing, the viscosity of the dough may be adjusted by additionally using 330 to 700 parts by weight of distilled water based on 100 parts by weight of the water-soluble polymer, but is not necessarily limited thereto.

Step (ii) is a step of processing into beads, and it is preferable to process the beads with an average particle size of 3 to 10 mm. If the average particle diameter of the beads is less than 3 mm, false positives may occur if a large amount of urine passing through the bentonite touches the beads, and there is a limit that it is difficult to visually check the color change. It is good to manufacture beads within the above range because there is a limit that a cat, which is characterized by being sensitive, may feel a foreign body feeling when stepping on a bead and refuse to use the toilet.

The step (iii) is to dry the beads to prepare solid beads, and the drying conditions may be performed at a temperature of 70 to 90 ° C for 1 to 3 hours, but are not necessarily limited thereto.

In addition, the present invention can provide a kit for diagnosing feline proteinuria comprising the solid beads and the bentonite coagulated sand.

In the present invention, "diagnosis" refers to determining a subject's susceptibility to a specific disease or disorder, determining whether an individual has a specific disease or disorder, and determining the prognosis of an individual suffering from a specific disease or disorder. or therametrics (eg, monitoring the condition of an individual to provide information about the efficacy of a treatment).

In addition, the present invention comprises the steps of mixing the solid beads with alkaline coagulation type sand; And confirming the color change when the cat's urine comes into contact with the solid beads; can provide a method for diagnosing proteinuria, including. At this time, the mixing of the solid beads with the alkaline coagulation-type sand may include both placing the beads on the surface of the alkaline coagulation-type sand or mixing them.

Therefore, the present invention uses a solid bead containing an acid solution together with a tetrabromophenol blue indicator for the accuracy of the diagnosis of proteinuria, so that when proteinuria is tested in alkaline coagulated sand such as bentonite, strong alkalis such as bentonite It is possible to accurately diagnose proteinuria by improving the color change errors.

Hereinafter, examples will be described in detail to aid understanding of the present invention. However, the following examples are merely illustrative of the contents of the present invention, but the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those skilled in the art.

<Production Example>

Materials used in the experiment were prepared in the following manner.

① Acid solution: Dissolve benzoic acid (5 mM, 15 mM, 25 mM), citric acid (5 mM, 15 mM, 25 mM, 35 mM), and tartaric acid (5 mM, 15 mM, 25 mM, 35 mM) in distilled water, respectively. An acid solution of was prepared.

② Tetrabromophenol blue indicator: A tetrabromophenol blue solution was prepared by dissolving 0.04 g of a commercially available tetrabromophenol blue reagent (product name: Tetrabromophenol blue) having a concentration of 85% in 30 ml of a 100% ethanol solution.

③ Water-soluble polymer: 100 g of microcrystalline cellulose was prepared.

2 g of the previously prepared acid solution and 30.04 g of a solution in which tetrabromophenol blue was dissolved were mixed, and 100 g of microcrystalline cellulose was added thereto and dispersed. After kneading them, they were put into a mold to prepare solid beads having an average particle size of 5 mm. The prepared solid beads were dried at 80° C. for 2 hours to prepare final solid beads. Figure 3 shows the appearance of a solid bead prepared according to the present invention.

<Comparative Example> Conventional proteinuria test method

The basic principle for measuring proteinuria changes color to blue by reacting tetrabromophenol blue with proteinuria as shown in FIG.

In order to confirm whether proteinuria can be tested with tetrabromophenol blue in bentonite, which is widely known as cat litter, the present inventors sprayed tetrabromophenol blue on a cat litter product (product name: PETHROOM DUST FREE BENTONITE) and tested the cat (female, age: 11). ) The scene for confirming the result by sprinkling again 5ml of urine collected from and observing the color change is shown in FIG.

As a result of FIG. 3, it was confirmed that bentonite showed a blue color change while showing an error reaction in distilled water, not proteinuria, because bentonite was alkaline in bentonite. That is, it was confirmed that the detection of proteinuria in bentonite with the conventional tetrabromophenol blue reagent was less accurate.

On the other hand, the indicator used in the existing silica gel sand was sprinkled on the bentonite in the same way as above, and distilled water was sprinkled thereon to observe the color change, and the results are shown in FIG. 4.

As a result of FIG. 4, it was confirmed that the color change to green was shown while indicating an error in the indicator used in the existing silica gel sand. This confirmed that the accuracy of the diagnosis of proteinuria was reduced with the indicator used in the existing silica gel sand in bentonite.

Taken together, these results confirm that it is difficult to detect proteinuria from cat urine in bentonite sand because bentonite is alkaline.

<Example> Confirmation of color change detection effect using solid beads: Confirmation of color change by type and concentration of acid solution

Through the previous experimental results, it was found that it was difficult to confirm the color change due to proteinuria from cat urine in bentonite sand.

The present inventors tried to detect proteinuria in bentonite sand in order to confirm the color change using solid beads prepared by varying the amount of acid solution for each type and concentration.

Specifically, benzoic acid 5 mM, 15 mM, 25 mM, citric acid 5 mM, 15 mM, 25 mM, 35 mM, tartaric acid 5 mM, 15 mM, 25 mM, 35 mM, respectively, were added to solid beads treated with cat (female). , Age: 5), 5 ml of urine collected from the sample was sprayed again, left for about 2 minutes, and color change was confirmed, and the results are shown in FIG. 6 .

As a result of FIG. 6, when observed with the naked eye, it was confirmed with the naked eye that the solid beads that did not contain an acid solution were slightly blue even in distilled water, and a clear color change was observed when tartaric acid was included in various acid solutions. could Furthermore, based on the results of FIG. 6, the difference in color change was objectively calculated numerically and shown in FIG. This is measured by measuring the RGB values of the cross section of the bead when it is negative and when it is positive, and then comparing the D value representing the distance from the reference value (1,1,1).

As a result of FIG. 7, benzoic acid showed a color change value of 2 to 25, citric acid a color change value of 8 to 44, and tartaric acid a color change value of 10 to 67. When 25 mM acid was included, the color change value for proteinuria was 67, indicating the most distinct color change. Through this, it was confirmed that tartaric acid, in particular, is the most suitable product because the color change for proteinuria is the most obvious.

Therefore, the present invention can accurately diagnose proteinuria by improving the error of color change due to strong alkali of bentonite during proteinuria test in bentonite sand by using solid beads containing an acid solution together with tetrabromophenol blue indicator. can

Having described specific parts of the present invention in detail above, it is clear to those skilled in the art that these specific descriptions are only preferred embodiments, and the scope of the present invention is not limited thereby. something to do. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (10)

(a) 100 parts by weight of a water-soluble polymer including cellulose or alginate or a mixture thereof,
(b) 0.04 to 0.1 parts by weight of tetrabromophenol blue indicator; and
(c) 1.2 to 5 parts by weight of an acid solution having a pH of 2 to 3.5;
A solid bead for diagnosing proteinuria in cat urine in alkaline coagulated sand, characterized in that it comprises a.
According to claim 1,
Characterized in that the cellulose is microcrystalline cellulose, a solid bead for diagnosing proteinuria in cat urine in alkaline coagulated sand.
According to claim 1,
The acid solution is at least one selected from the group consisting of benzoic acid, citric acid, and tartaric acid. body-shaped beads.
According to claim 1,
The solid bead is a solid bead for diagnosing proteinuria in cat urine in alkaline coagulated sand, characterized in that tetrabromophenol blue reacts with proteinuria in cat urine to induce a color change.
According to claim 4,
The color change is a solid bead for diagnosing proteinuria in cat urine in alkaline coagulated sand, characterized in that it has a discoloration range at pH 3.0 to 4.6.
According to claim 1,
The solid bead is characterized in that it further comprises at least one selected from the group consisting of cypress water, activated carbon, deodorant, phytoncide and cypress oil, a solid form for diagnosing proteinuria in cat urine in alkaline coagulated sand. bead.
(i) 0.04 to 0.1 parts by weight of a tetrabromophenol blue indicator based on 100 parts by weight of a water-soluble polymer including cellulose or alginate or a mixture thereof; and mixing and kneading 0.3 to 0.6 parts by weight of an acid solution having a pH of 2 to 3.5;
(ii) putting the dough into a molding machine and processing it into a bead shape; and
(iii) drying the bead-shaped workpiece;
Characterized in that it comprises, a method for producing solid beads for the diagnosis of proteinuria in cat urine.
According to claim 7,
The mixing in step (i) includes at least one solvent selected from the group consisting of ethanol, propanol, butanol, pentanol, hexanol, methanol, and dimethyl sulfoxide; Or distilled water; characterized in that further mixing, a method for producing solid beads for diagnosing proteinuria in cat urine.
Any one of the solid beads selected from claims 1 to 6; and alkaline coagulated sand, a diagnostic kit for feline proteinuria.
Mixing the solid beads of any one of claims 1 to 6 with alkaline coagulated sand; and
A method for diagnosing proteinuria comprising the step of checking the color change when the cat's urine contacts the solid beads.

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