JPH0819595A - Agent for preventing generation of ammonia - Google Patents

Abstract

PURPOSE:To provide an agent for preventing generation of ammonia from urea under the presence of urease by using an urease inhibitor extracted from Rscus acule atus, Psidium quayava, Rosa rugosa, Rubussuavissimus, Artemisia princeps, Tamarindus indica, and Eribotrya japanica or the like, as an effective component. CONSTITUTION:In enzyme urease produced by microorganism in excrements, urea in urine excreted from the human body reacts with water so as to produce ammonia and carbon dioxide gas, and thus produced ammonia increases the pH value of the skin so as to tend to cause diaper rash. Accordingly, natural urease inhibitor which is highly safe and easily usable, is extracted from Rscus acule atus, Psidium quayava, Rosa rugosa, Rubussuavissimus, Artemisia princeps, Tamarindus indica, and Eribotrya japanica or the like. Concentrate obtained by only removing an extracting solution from the this extract or a dry substance thereof is directly used as an ammonium generation preventing agent, or is dissolved in water, water-containing alcohol, 1,3-butylene glycol or the like so as to obtain a liquid solution or a power-like material.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an ammonia generation inhibitor which prevents urea from producing ammonia in the presence of urease.

[0002]

BACKGROUND OF THE INVENTION Urease, an enzyme produced by microorganisms in feces, is an enzyme that catalyzes the reaction of urea in the urine excreted outside the body with water to produce ammonia and carbon dioxide. When human or livestock manure is left in the environment, urea in the urine causes the above-mentioned enzymatic reaction, and the produced ammonia diffuses into the atmosphere, causing a strong unpleasant odor.

When the above-mentioned enzymatic reaction occurs before exchanging a diaper soiled with manure, the produced ammonia raises the pH of the skin, thereby enhancing the activity of protease and lipase and inducing diaper rash. Has been.

Therefore, in the case where ammonia may be produced from urea, preventing urease from acting so as to prevent the generation of ammonia is an effective means for preventing a bad smell and diaper rash caused by ammonia. As a substance having an urease inhibitory action that can be used for that purpose, conventionally known are hydroxam amphoteric surfactants, phosphoric acid amide derivatives, citric acid, malic acid, fumaric acid, phosphoric acid and the like.

[0005]

DISCLOSURE OF THE INVENTION The object of the present invention is to find a natural product type urease inhibitor which is highly safe and easy to use, and provides a novel ammonia generation inhibitor comprising the urease inhibitor. It is to facilitate the prevention of offensive odors from animal manure and the prevention of diaper rash.

[0006]

SUMMARY OF THE INVENTION Ammonia generation inhibitors successfully provided by the present invention include rucas, guava,
Maikai, Rubus, Wormwood, Tamarind, Yukinoshita,
The active ingredient is a urease inhibitor extracted from loquat or persimmon.

[0007] The term "ruscus" as used herein refers to the lily plant Rscus a
It means the underground part of cule atus. Guava means the leaves of Psidium guayava. Maikai means the flower part of Rosa rugosa. Rubus means the leaves of Rubussuavissimus. Mugwort means the aerial part of the Asteraceae plant Artemisia princeps. Tamarind is a legume Tamar
indus means indica seed coat. Yukinoshita means the aerial part of the Saxifraga stolonifera plant. Loquat is a Rosaceae plant Eriobotrya japonica
Means the leaves. In addition, persimmon means Dio
It means the leaves of spyros kaki.

Urease inhibitors which are effective in preventing the generation of ammonia include rucas, guava, maikai, rubus, mugwort, tamarind, yukinoshita, loquat, or persimmon, lower aliphatic alcohol, lower alkyl ketone, lower fatty acid ester. , A halogenated hydrocarbon, water, and a mixture thereof, and then extracted with an extraction solvent (though the solvent usable for the extraction is not limited thereto).

The extraction may be carried out by immersing the raw material plant which has been dried for easy treatment and ground, if necessary, in an extraction solvent at room temperature or in a warm state, but any other extraction method may be adopted. be able to. Specific examples of preferable extraction solvents include methanol, ethanol, butanol, acetone, methyl ethyl ketone, ethyl acetate, methylene chloride, water and the like. The hydrophilic organic solvent can be used as a mixture with up to about 20 times the amount of water. Among these, methanol, ethanol, butanol, acetone,
Alternatively, a mixture of these and water is particularly preferable because it gives an extract having a strong urease inhibitory activity in a high yield.

The main body of the urease inhibitory substance extracted from the above-mentioned plants such as Ruscus has not been confirmed yet, but it is presumed to be a polyphenolic compound.

An extract containing a urease inhibitor is
The concentrate obtained by simply distilling off the extraction solvent or its dried product can be used as it is as the ammonia generation inhibitor of the present invention, but if necessary, any substance effective for decolorization, deodorization, or concentration of a urease inhibitor can be used. You may use it for an ammonia generation inhibitor, after performing a refining process. The urease inhibitory activity of the extract can be confirmed by the method described in Example 1 below.

The urease inhibitory substance extracted from Ruscus or the like or the above plant extract containing it is in the form of a solution dissolved in water, hydroalcohol, 1,3-butylene glycol or the like, or in the form of a powder. It is used as the ammonia generation inhibitor. When the solution is used, the concentration is not particularly limited, but when a plant extract containing a urease inhibitor is used, 0.0001 to 10% by weight is suitable, and a particularly preferable concentration is 0. 0.001-1.0
% By weight.

The ammonia generation inhibitor of the present invention may contain other urease inhibitors in addition to the above-mentioned plant-derived urease inhibitors. Further, when formulating the ammonia generation inhibitor of the present invention into a deodorant, a diaper rash inhibitor, etc., a substance having a deodorant action by a mechanism different from urease inhibition, such as green tea extract, Vladimirium vulgaris extract, You may use together chlorophyll, flavonoid, etc.

The ammonia generation inhibitor of the present invention can be used in any method wherever it is desired to prevent the formation of ammonia from urea in the presence of urease. For example, if it is desired to eliminate the ammonia odor in a litter box, livestock barn, pet animal breeding room or excretion site, etc., spray or spray it on or near the cause of the occurrence. It may be attached in advance to the straw of a barn or the excrement absorbent material of a pet animal. When used to prevent diaper rash due to ammonia generated from urea, it is attached to the diaper by a method such as spraying, dipping or coating.

[0015]

【Example】

Example 1 1000 g of 50% ethanol was added to 100 g of crushed rucus.
After adding ml, the mixture is allowed to stand at 40 ° C. for 24 hours to extract soluble components. The extract is concentrated to dryness under reduced pressure to obtain a dried product. The urease inhibitory activity of the obtained Ruscus extract is measured by the following method.

Urease inhibitory activity measurement method: 0.1% urea solution 20 μl urease solution (20 unit / ml, pH 7.0 phosphate buffer solution) 100 μl phenol-nitroprusside sodium test solution (Japanese Pharmacopoeia) 1.0 ml Ruscus extract solution 100 μl The above solutions were mixed well in a test tube and heated to 37 ° C for 15 minutes, then 1.0 ml of sodium hypochlorite / sodium hydroxide (Japanese Pharmacopoeia) was added and the mixture was further incubated at 37 ° C for 10 minutes.
Warm to. After the completion of the enzyme reaction and immediately after mixing the four types of solutions, the absorbance at a wavelength of 640 nm, which is proportional to the amount of indophenol produced from the sodium phenol-nitroprusside reagent, is measured.
As a control, the same operation and absorbance measurement are performed when water is added instead of the Ruscus extract solution.

From the measurement results, the inhibition rate of urease activity is calculated by the following formula. Inhibition rate (%) = [1- (A ₁ −A ₀ ) / (A ₃ −A ₂ )] ×
100 However, A ₀ : Absorbance in the case of adding the Ruscus extract; before the initiation of the enzymatic reaction A ₁ : Absorbance in the case of adding the Ruscus extract; After the completion of the enzymatic reaction A ₂ : Control; Before the initiation of the enzymatic reaction A ₃ : Control After completion of enzymatic reaction

The above-mentioned measurement is carried out with respect to the solution of the rucus extract having various concentrations, and the concentration of rucus extract and the IC ₅₀ value at which the inhibition rate is 50% are determined. The same raw material is subjected to the same extraction treatment by changing the extraction solvent, and the IC ₅₀ value of the obtained extract is determined.

The preparation of the extract of luscus and the measurement of the urease inhibitory activity were carried out by the above-mentioned method, and the preparation of the extract and the inhibition of urease of guava, maikai, rubus, mugwort, tamarind, yukinoshita, loquat and persimmon were also carried out in the same manner. The activity was measured. The results are shown in Tables 1-9.

[0020]

[Table 1] Ruscus extract Extraction solvent Extraction yield (wt%) Extract IC ₅₀ (μg / ml) Water 13.2 690 Water / ethanol mixture (1/1) 30.0 550 Ethanol 32.6 500 Methanol 34.3 520 Water / methanol mixture (1/1) 30.8 550 Acetone 24.8 730 Water / acetone mixture (1/1) 26.1 490 n-Butanol 8.2 880 Ethyl acetate 2.5 900 Chloride Methylene 0.4 950

[0021]

[Table 2] Guava extract Extraction solvent Extraction yield (wt%) Extract IC ₅₀ (μg / ml) Water 9 320 Water / ethanol mixture (1/1) 17 140 Ethanol 7 200 Methanol 10 190 Water / methanol Mixed liquid (1/1) 19 110 Acetone 8 380 Water / acetone mixed liquid (1/1) 14 200 n-Butanol 6 590 Ethyl acetate 4 890 Methylene chloride 1 950

[0022]

[Table 3] Maikai extract Extraction solvent Extraction yield (wt%) Extract IC ₅₀ (μg / ml) Water 20 410 Water / ethanol mixture (1/1) 23 150 Ethanol 10 230 Methanol 20 210 Water / methanol Mixed liquid (1/1) 29 290 Acetone 6 330 Water / acetone mixed liquid (1/1) 14 280 n-Butanol 4 720 Ethyl acetate 2 970 Methylene chloride 2 890

[0023]

[Table 4] Rubus extract Extraction solvent Extraction yield (wt%) Extract IC ₅₀ (μg / ml) Water 11 510 Water / ethanol mixture (1/1) 31 300 Ethanol 25 280 Methanol 21 320 Water / methanol Mixed liquid (1/1) 33 290 Acetone 19 450 Water / acetone mixed liquid (1/1) 21 350 n-Butanol 6 690 Ethyl acetate 3 730 Methylene chloride 1 790

[0024]

[Table 5] Artemisia extract Extract solvent Extract yield (wt%) Extract IC ₅₀ (μg / ml) Water 12 1030 Water / ethanol mixture (1/1) 29 470 Ethanol 27 560 Methanol 26 600 Water / methanol Mixed liquid (1/1) 30 530 Acetone 13 750 Water / acetone mixed liquid (1/1) 25 520 n-Butanol 10 920 Ethyl acetate 3 1320 Methylene chloride 0.8 1640

[0025]

[Table 6] Tamarind extract Extraction solvent Extraction yield (wt%) Extract IC ₅₀ (μg / ml) Water 13 390 Water / ethanol mixture (1/1) 30 100 Ethanol 33 200 Methanol 34 120 Water / methanol Mixed liquid (1/1) 21 100 Acetone 23 310 Water / acetone mixed liquid (1/1) 30 120 n-Butanol 16 430 Ethyl acetate 9 800 Methylene chloride 0.4 720

[0026]

[Table 7] Yukinoshita extract Extraction solvent Extraction yield (wt%) Extract IC ₅₀ (μg / ml) Water 14 500 Water / ethanol mixture (1/1) 15 280 Ethanol 4 420 Methanol 6 460 Water / methanol Mixed liquid (1/1) 16 300 Acetone 4 540 Water / acetone mixed liquid (1/1) 11 290 n-Butanol 3 640 Ethyl acetate 2 740 Methylene chloride 1 770

[0027]

[Table 8] Loquat extract Extraction solvent Extraction yield (% by weight) Extract IC ₅₀ (μg / ml) Water 12 330 Water / ethanol mixture (1/1) 15 290 Ethanol 9 310 Methanol 11 300 Water / methanol Mixed liquid (1/1) 19 240 Acetone 7 530 Water / acetone mixed liquid (1/1) 13 230 n-Butanol 6 600 Ethyl acetate 3 790 Methylene chloride 0.9 730

[0028]

Table 9 Persimmon extract Extraction solvent Extraction yield (wt%) Extract IC ₅₀ (μg / ml) Water 10 190 Water / ethanol mixture (1/1) 10 170 Ethanol 9 210 Methanol 11 240 Water / methanol Mixed liquid (1/1) 15 150 Acetone 6 420 Water / acetone mixed liquid (1/1) 11 180 n-Butanol 5 580 Ethyl acetate 2 740 Methylene chloride 2 880

Example 2 A water / ethanol mixed liquid extract of each plant according to Example 1 was dissolved in a 10% aqueous ethanol solution to prepare a solution having a concentration of 1000 ppm, 50 ml of which was 10
Sprayed on 0 g of straw. The treated straw was air-dried and then put in a plastic vat, and 500 ml of artificial urine (urea 1.7%, salt 1.5%, pH 6.0) was mixed with 200 units of urease and poured. After storing at room temperature for 3 days, the ammonia concentration in the space above the straw was measured.

As a control, the same measurement was carried out when artificial urine and urease were poured in the same manner except that 50 ml of a 10% aqueous ethanol solution containing no water / ethanol mixed liquid extract was first sprayed.

As a result, 500 ppm of ammonia was detected in the control group, whereas in the test group in which the solution of the water / ethanol mixed liquid extract had been sprayed in advance, ammonia was detected in all the plant extracts. Was not detected (detection limit 10 ppm).

In addition to the measurement of the ammonia concentration, 10
A sensory test was conducted by a panel of representatives, and all 10 people in the control area felt the unpleasant odor of ammonia, whereas the test area 1 person did not feel the unpleasant odor of ammonia, and a slight ammonia odor. There were 9 people who felt.

Example 3 The rooibos aqueous extract according to Example 1 was dissolved in a 10% aqueous solution of ethanol to prepare a solution having a concentration of 1000 ppm, which was used as a cotton diaper and 10 parts per diaper (about 50 g).
It was applied at a ratio of ml and dried. This rooibos extract-treated diaper was worn by 10 infants for 4 weeks. Untreated diapers were worn by 10 people in the control group under the same conditions.

After the end of the test period, diaper rash was observed in 3 out of 10 people in the control group, but no rash was found in 10 of the diaper wearing section treated with rooibos extract.

[0035]

The ammonia generation inhibitor of the present invention is highly safe because it comprises a plant-derived urease inhibitor.
Since it is considered to have good biodegradability, it can be used at any time to prevent the generation of offensive odors from manure and diaper rash without adversely affecting the environment.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI technical display location C12N 9/99

Claims (2)

[Claims] 1. Ruscus, guava, Maikai, Rubus,
An ammonia generation inhibitor comprising a urease inhibitor extracted from mugwort, tamarind, yukinoshita, loquat, or persimmon. 2. Ruscus, guava, Maikai, Rubus,
Mugwort, tamarind, yukinoshita, loquat, or persimmon,
It is characterized by containing an extract obtained by extraction treatment with an extraction solvent selected from the group consisting of lower aliphatic alcohols, lower alkyl ketones, lower fatty acid esters, halogenated hydrocarbons, water, and mixtures thereof. Ammonia prevention agent.