JP2010220641A - beta-GLUCURONIDASE INHIBITOR - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an agent for persistently restraining the generation of malodor derived from urine by making clear malodor component more highly contributing urine odor than ammonia in a living environment and inhibiting enzyme related to the generation. <P>SOLUTION: New β-glucuronidase inhibitor is selected from creeping saxifrage, Damask rose, uva ursi, lianous Saururaceae, Henna (colorless), Rhododendron dauricum, tree peony and so on and extracts thereof. An environmental hygienics product and a sanitary product contain the β-glucuronidase inhibitor. A composition for restraining the generation of urine odor and a method of restraining the generation of urine odor use the β-glucuronidase inhibitor. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a β-glucuronidase inhibitor and a composition for suppressing urine odor production.

  In recent years, it has been strongly desired to remove not only the appearance of dirt but also the odor reminiscent of the presence of dirt due to the increase in consumer hygiene. Especially for urine and feces, the existence cannot be separated from the living environment, and furthermore, the odor strongly reminds the excrement itself. In toilets, stool can be easily discharged outdoors by washing with water, but a small amount of urine remains outside the toilet as droplets, and its presence is difficult to visually check, so it remains on the spot for a long time. , Easy to cause odor. In addition, sanitary products such as underwear, diapers, and sanitary products may exist in the living environment for a certain period of time with urine attached, and can be a source of malodor originating from urine.

  Conventionally, as a general deodorizing technique against such a bad odor, a deodorant that physically or chemically adsorbs the bad odor component has been used. As an effect having such an effect, the effect of green tea extract is widely known as a plant extract.

  On the other hand, it is generally considered that the odor of urine immediately after urination is very weak, and most of the malodorous components derived from urine are generated with the passage of time due to the action of metabolic enzymes derived from microorganisms. For this reason, development of the technique which can suppress continuously the production | generation of the malodorous component from urine is desired.

  Such urine-derived malodorous components include ammonia produced from urea by urease, and as its production suppression technology, ammonia generation suppression technology using a urease activity inhibitor has been proposed (for example, Patent Documents 1 to 3). 4).

  However, because ammonia has a high threshold as a bad odor component (it does not feel odor unless it is high in concentration), nowadays the excrement is immediately discharged to the outdoors due to the widespread use of the flushing method. Scenes that feel strong are very rare.

  On the other hand, β-glucuronidase is an enzyme that hydrolyzes glucuronide-conjugated compounds (glucuronides) with various alcohols, phenols, amines, etc., and is present in many organisms such as bacteria, fungi, plants, and animals. For example, it is known that a steroid compound secreted by glucuronide conjugation in human sweat is involved in the generation of glandular odor through metabolism of bacteria living in the skin (Patent Literature) 5) The relationship between the enzyme and urine odor is not known.

JP 2006-255290 A JP 2004-91338 A Japanese Patent Laid-Open No. 5-137774 JP 2006-192127 A Japanese Patent Laid-Open No. 2002-255776

  Therefore, the object of the present invention is to clarify the malodorous component that contributes more to urine odor than ammonia in the living environment, and to inhibit the generation of odor derived from urine by inhibiting the enzymes involved in its generation. It is to provide an agent that can be used.

  The present inventors have found that phenolic compounds and indoles generated from urine are components that have a high contribution to urine odor, and that these components are notable due to the action of β-glucuronidase derived from bacterial cells on urine. Found to increase. In addition, ammonia odor is rarely felt strongly in the living environment where current excrement is immediately excreted outdoors, and the urine odor generally felt by consumers is lower than that of ammonia. The above phenolic compounds and indoles are considered to be malodorous components.

  The inventors of the present invention have the advantage that a specific plant extract has an excellent β-glucuronidase inhibitory effect, and can further suppress the generation of unpleasant urine odor by these β-glucuronidase inhibitors, Has been found to be able to impart urine odor production inhibiting effects to these products.

  The present invention relates to saxifrage, damask rose, walrus, crested damselfly, henna (carales), Ezomura red azalea, buttons, itadori, maregale, rosa kentifolia, oysters, budgerigars, neutral henna, red snapper, american maple, hermanus, tartary buckwheat, Japanese yam, honey beetle, sorghum, lemon eucalyptus, oak (myrobaran), ganbeioki, tonkin nikkei, nagagamishigishi, aurora, dog larch, mistletoe, enoki bean, gyoju, hamasuge, eucalyptus, litchi, white-bellied sorghum, sorghum Whether it is emberia, dogwood, hornworm, mugwort, clove, mushroom, bracken, suou, taiwansendan, and their extracts There is provided a β- glucuronidase inhibitor selected.

  The present invention also provides a composition for suppressing urine odor production comprising the above novel β-glucuronidase inhibitor as an active ingredient. Furthermore, the present invention provides a method for suppressing the generation of urine odor, wherein the β-glucuronidase inhibitor is applied before urine adheres to an object or before drying after urine has adhered.

  The present invention also provides an environmental hygiene product and a sanitary product imparted with a urine odor production inhibitory effect by containing the β-glucuronidase inhibitor described above.

  Since the β-glucuronidase inhibitor of the present invention can continuously suppress the generation of malodorous components that characterize urine odors, it has excellent deodorizing effects, environmental hygiene products such as deodorants, diapers, sanitary products, It is useful in sanitary products such as animal excrement-related products such as pet excretion sheets for dogs and cats. The β-glucuronidase inhibitor of the present invention is a means for solving various problems other than urine odor caused by the action of β-glucuronidase, such as a body odor production inhibitor derived from volatile steroids, bladder cancer or colon cancer. It can also be used as a drug or a food that reduces the occurrence of.

It is a sensory evaluation result which shows the increase in the urine odor intensity | strength by (beta) -glucuronidase. It is a figure which shows the measurement result of (beta) -glucuronidase activity of sterilized urine, rot urine microbial cells, and rot urine supernatant. It is a figure which shows the sensory evaluation result of the urine odor intensity | strength of sanitized urine and rot urine. It is a figure which shows the urine odor production inhibitory effect of the plant extract with respect to (beta) -glucuronidase addition urine. It is a figure which shows the beta-glucuronidase inhibitory effect of the plant extract with respect to a urine odor production microbe. It is a figure which shows the urine odor production inhibitory effect of the plant extract with respect to urine odor production microbe addition urine.

[Β-glucuronidase inhibitor]
β-glucuronidase is an enzyme that hydrolyzes a compound (glucuronide) in which various alcohols, phenols, amines and the like are conjugated with glucuronide, and is present in many organisms such as bacteria, fungi, plants, and animals. In the present invention, β-glucuronidase derived from bacteria and fungi is important because microorganisms are greatly involved in the degradation of urine excreted outside the body. Specific examples include β-glucuronidase derived from Escherichia coli, Lactobacillus brevis, Propionibacterium acnes, Clostridium perfringens, Staphylococcus haemolyticus, Streptococcus agalactiae, Streptococcus pyogenes, Haemophilus somunus, Shigela sonnei, Aspergillus niger and the like. Β-glucuronidases derived from these microorganisms are classified into enzyme groups having a common domain. Furthermore, β-glucuronidase derived from human plasma is also classified into a similar protein group.

  In the β-glucuronidase inhibitor used in the present invention, the activity of 1.6 units / mL E. coli-derived β-glucuronidase Type VII-A was suppressed by 80% or more by adding 0.0025% by weight to the reaction solution. Refers to those marked with Furthermore, urine, stool, or a mixture of urine and stool from an animal such as a human, preferably urine, stool, or a mixture of urine and stool attached to an object such as a toilet environment such as a toilet bowl or toilet floor or a sanitary product, It also has an activity-inhibiting effect on β-glucuronidase active bacteria extracted from objects such as toilets such as toilets and toilet floors that have adhered or contacted with them or sanitary products, especially β-glucuronidase derived from Escherichia coli. What is shown is preferred.

  Hereinafter, among the β-glucuronidase inhibitors defined as described above, the novel β-glucuronidase inhibitors found in the present invention are listed.

  Yukinoshita, damask rose, walrus, tsudokudami, henna (carales), ezoramaki azalea, button, itadori, maregale, rosa kentifolia, oysters, budgerigars, neutral henna, red snapper, corn borer, hermanus, tartary dragonfly, clover Sorghum, Lemon Eucalyptus, Oak (Mirobaran), Gambiroki, Tonkin Nikkei, Nagabagishigishi, Fuso, Inu Larix, Mistletoe, Enoki Beans, Gyory, Hamasuge, Eucalyptus, Reishi, White-bellied Giraffe, Sequoia serrata, Syria Goldworm, mugwa, mushroom, firewood, suou, taiwansendan, and extracts thereof.

  One or more of the whole plant, leaves, roots, rhizomes, fruits, seeds and flowers of the plant can be used as it is or after being pulverized. When it is set as an extract, it can obtain by solvent-extracting the said plant or fungi, or its ground material at normal temperature or a heating. In the extraction, an extraction device such as a Soxhlet extractor can be used.

  Solvents used for extraction include water; alcohols such as methanol, ethanol and propanol; polyhydric alcohols such as propylene glycol, butylene glycol and glycerol; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate and ethyl acetate. Chain and cyclic ethers such as tetrahydrofuran and dimethyl ether; halogenated hydrocarbons such as dichloromethane, chloroform and carbon tetrachloride; hydrocarbons such as hexane, cyclohexane and petroleum ether; aromatic hydrocarbons such as benzene and toluene; Polyethers such as polyethylene glycol; pyridines and the like can be mentioned, and these can be used alone or as a mixture. A supercritical fluid such as carbon dioxide can also be used.

  Among the above extraction solvents, a solvent selected from water, alcohols such as ethanol, polyhydric alcohols such as butylene glycol, or a mixed solvent of two or more of these is preferable, and a mixed solvent of water and ethanol may be used. preferable. The volume ratio of the mixed solvent of water and ethanol (water: ethanol) is preferably 10:90 to 90:10, and more preferably 60:40 to 40:60.

  The obtained various solvent extracts can be used as they are, and can also be used as a diluted solution or concentrated solution, or after being concentrated or lyophilized, in the form of powder or paste. Moreover, it is also possible to remove inactive impurities from the extract by a technique such as liquid-liquid distribution, and it is preferable to use such a material in the present invention. These may be used after performing treatments such as deodorization and decolorization by a known method, if necessary.

[Composition for inhibiting urine odor production, method for inhibiting urine odor production, environmental hygiene products, sanitary products]
In the present invention, any of the above β-glucuronidase inhibitors can be contained alone or in combination of two or more in the composition for suppressing urine odor production. Further, by containing an effective amount of the β-glucuronidase inhibitor or urine odor production-suppressing composition of the present invention in an environmental hygiene product or sanitary product for home use or facility use, the production of malodor derived from urine is prevented. Thus, a product having a high suppression effect and deodorization effect can be obtained. In addition, it is preferable that the β-glucuronidase inhibitor is added to the target object (thing to suppress the generation of urine odor) before urine adheres or after adhering urine and before the urine is dried. By applying within 1 hour, more preferably within 10 minutes after adhering, the generation of urine odor can be effectively suppressed.

  The urine is not limited to those derived from humans, and may be urine derived from animals such as pets such as dogs and cats. The malodor derived from urine is not limited to the malodor generated from urine alone, and may be a malodor generated from a state in which urine and feces are mixed in a diaper, for example.

  Malodorous components that can be controlled by the β-glucuronidase inhibitor of the present invention are phenolic compounds and indoles, specifically phenol, p-cresol, 4-vinyl-2-methoxy-phenol, 4-vinylphenol, Examples include, but are not limited to, 2-methoxy-1,3-benzenediol, 1,4-benzenediol, 1,3-benzenediol, indole, and volatilization caused by the action of β-glucuronidase on urine. What is necessary is just a sex component.

  The content of the β-glucuronidase inhibitor in the composition for suppressing urine odor production of the present invention is such that the concentration of the β-glucuronidase inhibitor is 0.0001 to 5% by weight at the time of contact mixing of the composition and the urine component. It is preferable to make it 0.001 to 1% by weight, and further 0.005 to 0.5% by weight.

(Environmental hygiene products)
Examples of environmental sanitary products include cleaning agents, deodorants, textile detergents, wipes and the like. Moreover, it is also possible to use several usage methods in combination. The environmental hygiene product of the present invention controls the generation of urine odor by applying it to places where urine-derived malodors are likely to occur, such as toilet floors, wall surfaces, toilets, waste containers, nursing homes, clothing, underwear, and bedding. can do.

  When the β-glucuronidase inhibitor or urine odor production suppressing composition of the present invention is used as a spray-type deodorant, the content as the β-glucuronidase inhibitor is preferably 0.001 to 10% by weight. Further, it is preferable to be 0.005 to 5% by weight, further 0.01 to 1% by weight.

  The composition for suppressing urine odor production used in the environmental hygiene product of the present invention is a variety of commonly used components such as oils, surfactants, alcohols, chelating agents, pH adjusters, bactericides, antibacterial agents, and antiseptics. In addition to thickeners, pigments, fragrances, etc., the present invention includes components such as deodorant bases, moisturizers, softeners, keratin protective agents, medicinal agents, antioxidants, solvents, metal salts and metal ions. In the range which does not inhibit the effect of, it can mix | blend arbitrarily and can formulate.

(Sanitary products)
Examples of sanitary products include absorbent articles such as light incontinence items, disposable paper diapers, urine-absorbing pads, sanitary items, and animal excrement-related items such as pet excretion sheets for dogs, cats, etc. Can be mentioned. The sanitary product of the present invention can control the generation of urine-derived malodor that occurs over time from adhering urine.

When used for absorbent articles, the β-glucuronidase inhibitor or urine odor production-suppressing composition of the present invention is preferably impregnated into a backing sheet or absorbent layer, particularly a water-absorbing polymer. impregnation amount of the β- glucuronidase inhibitor is preferably a 0.005~10g / m ² with respect to the application surface, more preferably between 0.05 to 2 g / m ^2.

  In the composition for suppressing urine odor production used in the sanitary product of the present invention, various commonly used components such as oil, surfactant, antistatic agent, antioxidant, pH adjuster, smoothing agent, antibacterial agent, In addition to antifungal agents, preservatives, pigments, fragrances, etc., components such as deodorant bases, medicinal agents, solvents, metal salts and metal ions are arbitrarily added as long as the effects of the present invention are not impaired. be able to.

(Deodorant base)
When the β-glucuronidase inhibitor of the present invention or the composition for suppressing urine odor production containing the same is used for environmental hygiene products or sanitary products, it is generally known as long as the β-glucuronidase inhibitory effect is not lost. A deodorant base can be used in combination or contained.

As an example of the above deodorant base,
Metal compounds such as iron oxide, iron sulfate, iron chloride, zinc oxide, zinc sulfate, zinc chloride, silver oxide, copper oxide;
Acids such as phosphoric acid, citric acid and succinic acid, and triethanolamine, monoethanolamine, sodium hydroxide, potassium hydroxide, 2-amino-2-hydroxymethyl-1,3-propanediol (tris (hydroxymethyl) An acid or a salt thereof having a pH buffering effect comprising a combination with a base such as aminomethane);
Carboxylic acids such as lactic acid, succinic acid, malic acid, citric acid, maleic acid, malonic acid;
Fatty acid metals such as zinc undecylenate, zinc 2-ethylhexanoate, zinc ricinol;
Plant extract-type deodorants such as catechin, polyphenol, green tea extract, mushroom extract, wood vinegar and bamboo vinegar
Metal type chlorophyllin sodium such as iron and copper, metal phthalocyanine such as iron, copper and cobalt, catalyst type such as tetraphthalic acid phthalocyanine such as iron, copper and cobalt, titanium dioxide, visible light responsive titanium dioxide (nitrogen doped type, etc.) Deodorants;
cyclodextrins such as α-, β-, or γ-cyclodextrin, its methyl derivatives, hydroxypropyl derivatives, glucosyl derivatives, maltosyl derivatives;
Alkylene glycols that are said to have a retention of malodor such as ethylene glycol, propylene glycol, diethylene glycol, dipropylene glycol;
Ester oil agent that is said to have an effect of retaining bad odor such as myristic acid esters, palmitic acid esters, phthalic acid esters, adipic acid esters, sebacic acid esters, citrate esters;
Acrylic acid polymer such as porous methacrylic acid polymer and porous acrylic acid polymer, porous divinylbenzene polymer, porous styrene-divinylbenzene-vinylpyridine polymer, aromatic polymer such as porous divinylbenzene-vinylpyridine polymer, and copolymers thereof Synthetic porous polymers such as chitin, chitosan and other natural porous polymers;
Inorganic porous materials such as activated carbon, silica, silicon dioxide (silica gel), calcium silicate, high silica zeolite (hydrophobic zeolite), sepiolite, cancrinite, zeolite, hydrated zirconium oxide;
Examples thereof include metal-supporting porous materials such as silver-supporting zeolite, silver-supporting cancrinite, and silver-supporting porous styrene-divinylbenzene-vinylpyridine polymer. These deodorant bases may be used alone or in combination.

(Organic solvent)
Furthermore, when using the β-glucuronidase inhibitor or urine odor production-suppressing composition of the present invention for environmental hygiene products or sanitary products, it is more preferable to use or contain a water-soluble organic solvent in combination.

Examples of the above water-soluble organic solvents include
Alcohols such as ethanol, 1-propanol, 2-propanol, butyl alcohol;
Ethylene glycol, 1,2-propanediol, 1,3-butanediol, hexylene glycol, 2-ethyl-1,3-hexanediol, glycerin, triethylene glycol, dipropylene glycol, diglycerin, polyethylene glycol, polypropylene glycol Polyhydric alcohols such as
And ethers such as ethylene glycol ethyl ether, ethylene glycol methyl ether, ethylene glycol butyl ether, and diethylene glycol ethyl ether.

Reference Example 1 Production of volatile compounds from urine by addition of β-glucuronidase
(1) Preparation of measurement sample Human urine sample (mixture of 5 human urine) that was sterilized with 0.2μm filter (Millipore Gyrex GV) immediately after collection in a γ-ray sterilized container ) Add 9.9mL, then add and mix 0.1mL of E. coli-derived β-glucuronidase Type VII-A (purchased from Sigma) adjusted to 250units / mL, leave it in a 25 ° C constant temperature bath and let it react for 22 hours. It was. What added 0.1 mL of sterilized ion-exchange water instead of the enzyme solution was similarly left as an initial urine sample in a thermostat for 22 hours.
After completion of the reaction, benzyl benzoate ethanol solution was added as an internal standard to 9 mL of the reaction solution, and extracted twice with 10 mL of diethyl ether. The extracts were combined and dried over anhydrous magnesium sulfate. After drying, the solid matter was filtered off, and the filtrate was concentrated with a rotary evaporator to obtain a measurement sample.

(2) Measurement of volatile compounds A gas chromatograph mass spectrometer was used for measurement. The production amount of the detected volatile compound was calculated as a peak area ratio with respect to the internal standard. The results are shown in Table 1.

  From these results, it was confirmed that various phenolic compounds and indole were produced from urine by the action of β-glucuronidase.

Reference Example 2 Generation of urine odor by adding β-glucuronidase
(1) Preparation of evaluation sample In a γ-ray sterilized container, 720 μL of a human urine sample (mixture of 5 human urine) that had been sterilized with a 0.2 μm filter immediately after collection was placed, followed by 16 units / 80 μL of β-glucuronidase aqueous solution adjusted to mL was added and mixed, and left in a 37 ° C. constant temperature bath to react for 20 hours. An initial urine sample was prepared by adding 80 μL of sterilized ion-exchanged water instead of the enzyme solution, and left in a constant temperature bath for 20 hours. About each sample, the equivalent amount was dripped at the front-end | tip of odor paper, and this was made into the evaluation sample.

(2) Odor sensory evaluation Odor sensory evaluation was performed by 6 panelists based on a 6-step odor intensity display method based on an evaluation score of 0 to 5 for odor intensity. Evaluation score is “0” odorless, “1” odor that can be finally detected (detection threshold), “2” urine odor, but weak odor (cognitive threshold), “3” odor that can be easily felt as urine odor “4” indicates a strong urine odor and “5” indicates a strong urine odor. Odor intensity was determined in 0.5 increments, and the average value of the evaluation is shown in FIG.
From this result, it was confirmed that the volatile component produced by the action of β-glucuronidase significantly increases the urine odor intensity.

Reference Example 3 Measurement of β-glucuronidase activity in spoiled urine
(1) Preparation of sample Septic urine was obtained by collecting 20 mL of a urine sample, dispensing it into a γ-ray sterilized container without sterilization, and storing it at room temperature for 7 days. 1 mL of this septic urine was centrifuged at 10,000 rpm for 5 minutes to separate the cells and supernatant. The precipitated cells were suspended in 0.5 mL of physiological saline. The supernatant was sterilized through a 0.2 μm filter.
Similarly, 15 mL of urine sterilized using a filter immediately after collection was also stored at room temperature for 7 days.

(2) Measurement of β-glucuronidase activity In a γ-ray sterilized container, 250 μL of 2 mM p-nitrophenyl-β-D-glucuronide (PNPG) aqueous solution, 100 μL of 0.5 M phosphate buffer (pH 6.8) and ion-exchanged water 100 μL was mixed, and then 50 μL of the urine sample (sterilized urine, rot urine cell suspension or rot urine supernatant) was added and reacted at room temperature.
Immediately after the start of the reaction and after the reaction for 24 hours, the reaction solution was diluted with 0.2 M glycine-sodium hydroxide buffer (pH 10.4), and the absorbance at a wavelength of 400 nm was measured. The value obtained by subtracting the absorbance immediately after the start from the absorbance after the reaction for 24 hours was used as an index of β-glucuronidase activity, and comparison was made for each sample. The results of activity measurement are shown in FIG.
From these results, it can be seen that cells grown in spoiled urine have β-glucuronidase activity, and supernatants that do not contain cells also have β-glucuronidase activity. It was confirmed that it was also secreted.

(3) Odorous sensory evaluation of sterilized urine and septic urine The sterilized urine and septic urine were subjected to odor evaluation at the bottle mouth with the containers used for storage.
Odor sensory evaluation was performed by 6 panelists based on the 6-step odor intensity display method shown in Reference Example 2 (2). The average of panelists' evaluation is shown in FIG.
From this result, it was confirmed that the bacterial cells growing in the urine increase the urine odor intensity.

Example 1 Inhibition of β-glucuronidase activity by plant extracts
(1) Preparation of plant extract solution 40 g of a tender eel plant base (dried product) was placed in a resin container and immersed in 400 mL of 50 vol% ethanol for 1 to 4 weeks to extract the components. The obtained various extracts were filtered (Whatman, qualitative filter paper) and concentrated with an evaporator. The obtained plant extract (solid matter) was dissolved in 50 vol% ethanol to adjust the solid content concentration to 1 w / v%. Extract solutions were prepared for other plants in the same manner.

(2) Relative activity inhibition rate measurement of β-glucuronidase 100μL of 2mM PNPG aqueous solution, 40μL of 0.5M phosphate buffer (pH6.8), 39.5μL of ion-exchanged water, and various plant extract solutions 0.5 in γ-sterilized containers Next, 20 μL of β-glucuronidase aqueous solution adjusted to 16 units / mL was added, and the enzyme reaction was carried out in a 37 ° C. constant temperature bath for 2 hours. The solid content concentration in the reaction solution of the plant extract provided is 0.0025 w / v%.
In addition, 50 vol% ethanol was added instead of the plant extract solution as a control, and each sample and control was added with ion exchange water instead of an enzyme solution as a blank. went.
As a comparison, also for D-glucaro-1,4-lactone, which is widely known to competitively inhibit β-glucuronidase, a 1 vol / vol% 50 vol% ethanol solution was prepared and reacted in the same manner. It was.
The reaction solution was diluted with 0.2 M glycine-sodium hydroxide buffer (pH 10.4), and the absorbance at a wavelength of 400 nm was measured. From the measured values obtained, the relative activity inhibition rate of β-glucuronidase was determined according to the following formula, and is shown in Table 2.

  From this result, it can be seen that the plant extract sample provided has β-glucuronidase inhibitory activity. The β-glucuronidase inhibitory activity preferably suppresses the above activity by 80% or more, and more preferably suppresses 90% or more.

Example 2 Hue Measurement of Plant Extract Solution (Gardner Color Number)
Since the plant extract solution often contains coloring components due to its nature, the plant extract solution is prepared according to the measurement method described in “JIS K0071-3 Color test method for chemical products, Part 2, Gardner color number”. The Gardner color number was measured. The results are shown in Table 3.

  As shown in Table 3, among the plant extract samples, the number of Gardner colors of tendered yak and oak (Mirobaran) is small, which is preferable when used alone or in combination with a product.

Example 3 Suppression of the generation of urine odor by a plant extract against urine containing β-glucuronidase
(1) Sample preparation 720μL of human urine sample (mixture of 5 human urine) and 1w / v% of various plants that were sterilized with 0.2μm filter immediately after collection in γ-ray sterilized containers Add 2 or 8 μL of the extract solution or green tea extract (Shirai Matsushin Pharmaceutical Co., Ltd., FS-1000) or 1 w / v% D-glucaro-1,4-lactone solution prepared to 1 w / v dry fraction. 80 μL of β-glucuronidase aqueous solution adjusted to 16 units / mL was added and mixed, and left in a 37 ° C. constant temperature bath to react for 20 hours. The concentration of various plant extracts, green tea extract and D-glucaro-1,4-lactone in the reaction solution is 0.0025 or 0.01% by weight.
In addition, 720 μL of the sterilized urine sample was mixed with 50 vol% ethanol 2 or 8 μL and the enzyme solution 80 μL, and the sterilized urine sample 720 μL was mixed with 50 vol% ethanol 2 or 8 μL and ion-exchanged water 80 μL. The added mixture was used as a blank and reacted for 20 hours in the same manner. About each sample, the equivalent amount was dripped at the front-end | tip of odor paper, and this was made into the evaluation sample.

(2) Confirmation of suppression of urine odor by sensory evaluation The odor sensory evaluation was performed by 6 panelists based on the 6-step odor intensity display method shown in (2) of Reference Example 2. The average paneler evaluation is shown in FIG.
From this result, it was shown that the plant extract having β-glucuronidase inhibitory activity found in the present invention has an excellent urine odor production suppressing effect.

Example 4 Inhibition of β-glucuronidase activity by plant extracts against urine odor producing bacteria
(1) Collection and identification of urine odor producing bacteria Surface materials in contact with the skin were collected from adult paper diapers worn by 4 panelists for 3 hours. Further, an equal amount of urine collected from the four panelists was mixed, and the recovered surface material was immersed therein. The urine sample thus prepared was absorbed in an adult paper diaper and stored in a constant temperature bath at 30 ° C. for 24 hours to generate a strong odor.
Strains (microbe colonies) were separated from this sample by a conventional method, and when these were cultured in sterilized urine, generation of a strong odor was confirmed for a specific strain. In the following evaluation, strains assigned to Escherichia coli by 16s rDNA partial base sequence analysis were used from these specific strains (urine odor producing bacteria).

(2) Preparation of bacterial solution The above isolated Escherichia coli is cultured in Mueller Hinton medium containing 3 mM PNPG and washed several times with physiological saline to remove the medium components, and Dulbecco's phosphate buffered saline (DPBS, Gibco To make the turbidity 0.5.

(3) Measurement of relative activity inhibition rate of β-glucuronidase Extraction of 2 mM PNPG aqueous solution 400 μL, 0.5 M phosphate buffer (pH 6.8) 160 μL, ion-exchanged water 160 μL, and 1 w / v% tender yam extraction in γ-ray sterilized containers 8 μL of a green tea extract (manufactured by Shiraimatsu Shinyaku Co., Ltd., FS-1000) or a 1 w / v% D-glucaro-1,4-lactone solution was mixed with a physical solution or a dry fraction of 1 w / v%, followed by the above 80 μL of the separated Escherichia coli solution prepared to a turbidity of 0.5 was added and mixed, and the reaction was performed in a 37 ° C. constant temperature bath for 2 hours. Concentrations in the reaction solution of the tender yam extract, green tea extract and D-glucaro-1,4-lactone are 0.01 w / v%.
In addition, a reaction in which 50 vol% ethanol was added instead of the tendi yak extract solution was used as a control, and a sample in which DPBS was added in place of the bacterial solution for each sample and control was used as a blank, and the reaction was similarly performed for 2 hours. The reaction solution was centrifuged at 10,000 rpm for 2 minutes, the supernatant was diluted with 0.2 M glycine-sodium hydroxide buffer (pH 10.4), and the absorbance at a wavelength of 400 nm was measured. From the obtained measured values, the relative activity inhibition rate of β-glucuronidase was determined according to the above formula, and the results are shown in FIG.
From these results, it was shown that the tender yam extract has an excellent β-glucuronidase inhibitory activity against urine odor producing bacteria isolated from the site of urine odor generation.

Example 5 Suppression of generation of urine odor by plant extract against urine added with urine odor producing bacteria
(1) Sample preparation 720μL of human urine sample (mixture of 5 human urine) that was sterilized with 0.2μm filter immediately after collection in a γ-ray sterilized container, and extracted with 1w / v% tendi yak A green tea extract (manufactured by Shiraimatsu Shinyaku Co., Ltd., FS-1000) adjusted to a 1% w / v dry fraction or 8 μL of a 1 w / v% D-glucaro-1,4-lactone solution was added, followed by Examples 80 μL of the separated Escherichia coli solution prepared to a turbidity of 0.5 shown in 4 (2) was added and mixed, and left in a 37 ° C. constant temperature bath for 4 hours to react. Concentrations in the reaction solution of the tender yam extract, green tea extract and D-glucaro-1,4-lactone are 0.01% by weight.
Similarly, 720μL of the sterilized urine sample mixed with 8μL of 50vol% ethanol and 80μL of the bacterial solution was used as a control, and 720μL of the sterilized urine sample was mixed with 8μL of 50vol% ethanol and 80μL of DPBS. For 4 hours.
The reaction solution was centrifuged at 10,000 rpm for 2 minutes, and the supernatant was sterilized using a 0.2 μm filter. About each sample, the equivalent amount was dripped at the front-end | tip of odor paper, and this was made into the evaluation sample.

(2) Confirmation of urine odor suppression by sensory evaluation The odor sensory evaluation was performed by 6 panelists based on the 6-step odor intensity display method shown in Reference Example 2 (2). The average of panelists' evaluation is shown in FIG.
From these results, it was shown that the tendered yak extract has an excellent urine odor production inhibitory effect against urine odor producing bacteria isolated from the site of urine odor generation.

Example 6 Spray-type Deodorant A composition for suppressing urine odor production containing a β-glucuronidase inhibitor according to the present invention having the composition (% by weight) shown in Table 4 was prepared, put in a trigger spray container, and sprayed. A mold deodorant was prepared.

Example 7 Disposable diaper A composition for suppressing urine odor production containing a β-glucuronidase inhibitor according to the present invention having the composition (% by weight) shown in Table 5 was prepared, and a disposable diaper was prepared using the composition. Specifically, the composition for suppressing urine odor production is sprayed on a disposable diaper mount (basis weight 16 g / m ² ) with a basis weight of 16 g / m ² , dried, and then used to mount 100 parts by weight of flap pulp. An absorbent body was obtained by wrapping a uniform mixture of 100 parts by weight of a water-absorbing polymer (polyacrylic acid crosslinked body). The basis weight of the flap pulp / water-absorbing polymer mixture in the absorbent was 300 g / m ² . A disposable diaper was prepared using the obtained absorbent body. At this time, the amount of the β-glucuronidase inhibitor in the composition for suppressing urine odor generation sprayed on the mount is 1.6 g / m ² .

Example 8 Urine collection pad A composition for suppressing urine odor production containing a β-glucuronidase inhibitor according to the present invention having the composition (% by weight) shown in Table 5 was prepared, and a urine collection pad was prepared using the composition. Specifically, only the water-absorbing polymer of the commercially available urine removing pad “Relief urine absorbing pad made by Kao” was changed to a water-absorbing polymer dried by adding and mixing the composition for suppressing urine odor production of the present invention. Was prepared. The addition amount of the composition for suppressing urine odor generation to the water-absorbing polymer was 10% by weight (before drying) with respect to the water-absorbing polymer. At this time, the addition amount of the β-glucuronidase inhibitor is 1% by weight with respect to the water-absorbing polymer, and is equivalent to 1.5 g / m ^{2 with} respect to the urine absorption pad.

Claims (5)

  Yukinoshita, damask rose, walrus, tsudokudami, henna (carales), ezoramaki azalea, button, itadori, maregale, rosa kentifolia, oysters, budgerigars, neutral henna, red snapper, corn borer, hermanus, tartary dragonfly, clover Sorghum, Lemon Eucalyptus, Oak (Mirobaran), Gambiroki, Tonkin Nikkei, Nagabagishigishi, Fuso, Inu Larix, Mistletoe, Enoki Beans, Gyory, Hamasuge, Eucalyptus, Reishi, White-bellied Giraffe, Sequoia serrata, Syria It is selected from the kinmizuhiki, mugwa (mulberry), crested cypress, bitumen, suou, taiwansendan, and their extracts - glucuronidase inhibitor.   An environmental hygiene product comprising the β-glucuronidase inhibitor according to claim 1.   A sanitary product comprising the β-glucuronidase inhibitor according to claim 1.   A composition for suppressing urine odor production, comprising the β-glucuronidase inhibitor according to claim 1.   A method for suppressing the generation of urine odor, wherein the β-glucuronidase inhibitor according to claim 1 is applied before urine adheres to an object or before drying after urine adheres.

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