CN109646361B

CN109646361B - Composite biological deodorization spray for pets and preparation method thereof

Info

Publication number: CN109646361B
Application number: CN201910143893.8A
Authority: CN
Inventors: 李静静; 李朝阳; 张思雨; 邵泽香; 刘学军; 乔彦良; 马广斌
Original assignee: Shandong Chongzhiyoupin Pet Food Co ltd; Shandong Zhuzi Biotechnology Co ltd; Shandong Sinder Technology Co ltd
Current assignee: Shandong Chongzhiyoupin Pet Food Co ltd; Shandong Sinder Technology Co ltd; Shandong Zhuzi Biotechnology Co ltd
Priority date: 2019-02-27
Filing date: 2019-02-27
Publication date: 2022-03-29
Anticipated expiration: 2039-02-27
Also published as: CN109646361A

Abstract

The invention relates to a composite biological deodorization spray for pets, which is suitable for solving the problems of dog and cat halitosis, body odor, excrement odor and the like, and a preparation method thereof, wherein the deodorization spray comprises, by mass, 5-8% of yucca extract, 4-5% of green tea extract, 6-8% of oregano oil extract, 4-5% of persimmon extract, 4-5% of horseradish extract, 3-4% of zinc ricinoleate, 5-6% of emulsifier, 1-2% of stabilizer, 1-2% of pH value regulator, and the balance of purified water. The deodorizing spray is finally verified by a deodorizing effect, and the deodorizing spray has good deodorizing performance, can quickly eliminate a plurality of common pet odor molecules such as ammonia gas, hydrogen sulfide, methyl mercaptan, methyl sulfide, dimethyl disulfide and the like within 5 minutes, can effectively inhibit and kill pathogenic microorganisms, and can radically inhibit the generation of odor.

Description

Composite biological deodorization spray for pets and preparation method thereof

Technical Field

The invention relates to a deodorant, and in particular relates to a composite biological deodorization spray for pets and a preparation method thereof.

Background

Bad breath, body odor and odor of fermented excreta of pets are a nuisance to every pet fan. The odor components are various and complex, so the removal difficulty is high, and the high-concentration odor not only influences the life quality of a pet owner, but also greatly influences the body health of the pet and the pet owner, such as the health of a respiratory system, an immune system, a digestive system, a mucous membrane and the like.

Most of the odor is generated due to the breeding of putrefying bacteria, which decompose organic matters such as carbohydrates, proteins and fats in pet excrement, oral cavity and skin, and generate odor molecules such as ammonia gas, hydrogen sulfide and thiols. The key point of radical treatment of odor is to kill putrefying bacteria and viruses such as harmful microorganisms, adsorb and dissolve odor in air, and decompose odor molecules through biological enzyme and catalyst, thereby achieving the purpose of radical treatment and deodorization.

The deodorant in the market is various in types, and mainly comprises a physical shielding deodorant, a chemical deodorant, an enzyme product deodorant, a plant deodorant and a microorganism deodorant. The deodorization methods all have advantages and disadvantages, wherein the physical shielding type deodorant is abundant, the odor is covered mainly by substances such as essence, the smell sense of the pet is very sensitive, and the odor is covered by the perfume but not a long-term and intelligent method, so that the deodorization effect is only temporary, can only treat symptoms and can not treat the root causes, can only temporarily cover the symptoms and can not be removed and maintained for a long time; the second kind of chemical deodorant represented by chlorine dioxide, hypochlorite, ferrous sulfate and the like mainly has the main effects of oxidation sterilization, the deodorization mechanism mainly comprises the reactions of desulfurization, oxidation, condensation, addition and the like on peculiar smell molecules, the sterilization products have strong sterilization effects, putrefying bacteria are eradicated, and the root source of odor generation is eradicated, so that the second kind of chemical deodorant has the auxiliary effect of serving as a deodorant, can treat the root, but the original odor in the air can not be rapidly removed, so that the second kind of chemical deodorant can treat the root and can not treat the symptoms, and the safety is poor; the plant deodorant is prepared by compounding extract liquor of oil, juice or extract extracted from roots, stems, leaves or flowers of plants according to a certain formula, wherein effective molecules in the plant extract contain active groups such as chemical activity, biological activity and conjugated double bonds, and can react with different odor substances such as acid-base reaction, catalytic oxidation reaction, Lewis acid-base reaction, redox reaction and the like to promote the odor substances to change the original molecular structure and decompose into nontoxic and tasteless small molecules, so that the aim of deodorization is fulfilled, if the single action has limited deodorization effect, only a part of odor molecules can be acted, and meanwhile, the cost is higher; the microorganism deodorant adopts deodorizing microorganisms obtained by screening to construct a compound microbial inoculum, has a better control effect on malodor sources consisting of perishable organic substances, and the deodorizing effect is not obvious probably because the microorganisms can only eliminate one or more malodor compounds, and if the eliminated compounds have poor correlation with the malodor intensity, the deodorizing effect of the microbial inoculum is not obvious.

In chinese patent application CN105724439A published in 2016, 7, 6, a disinfectant and bactericidal deodorant for pets is disclosed, which comprises dioctadecyl dimethyl ammonium chloride, didecyl dimethyl ammonium chloride, benzalkonium bromide, glutaraldehyde, pure plant extract, plant essential oil, and distilled water. The deodorant has remarkable sterilizing and deodorizing effects, and can be sprayed on pet cages, urine and feces, or sprayed in the air to the part needing sterilizing and deodorizing, to achieve the effects of sterilizing and deodorizing. However, the disinfectant and bactericidal action of the deodorant is that chemical disinfectant components are added, so that the safety cannot be guaranteed, and the deodorant is inconvenient to use because the deodorant needs to be diluted for use.

In chinese patent application CN108245449A published in 2018, 7 and 6, a deodorant for pets is disclosed, which contains linalool acetate, citronellol, phenethyl alcohol, parachlorometaxylenol, lauryl methacrylate, camphor extract, polysorbate, citric acid, essence, alcohol, deionized water. The deodorant can be directly sprayed on pet body, and has remarkable deodorizing effect, long lasting time, and antibacterial effect. However, the preparation process of the deodorant is complicated, resulting in an increase in production cost.

The invention is provided to solve the above problems.

Disclosure of Invention

The invention provides a composite biological deodorization spray for pets and a preparation method thereof, solves the problems of poor deodorization effect, temporary solution, non-permanent solution, no sterilization effect, poor safety, environmental hazard, high cost and the like of a market deodorization product for pets, improves the deodorization effect, improves the safety, has low cost, and is suitable for market popularization.

In order to achieve the purpose, the invention provides the following technical scheme:

a composite biological deodorization spray for pets comprises, by mass, 5-8% of Yucca extract, 4-5% of green tea extract, 6-8% of oregano oil extract, 4-5% of persimmon extract, 4-5% of horseradish extract, 3-4% of zinc ricinoleate, 5-6% of emulsifier, 1-2% of stabilizer, 1-2% of pH value regulator and the balance of purified water.

Preferably, the yucca extract comprises saponin, polysaccharide and polyphenol, wherein the content of saponin is 10-12%, the content of polysaccharide is 10-12%, and the content of polyphenol is 15-17%.

Preferably, the green tea extract comprises tea polyphenol, wherein the content of the tea polyphenol is 20-25%.

Preferably, the oregano oil extract comprises thymol, polyphenol and carvacrol, wherein the content of thymol is 10-11%, the content of polyphenol is 7-8% and the content of carvacrol is 8-10%.

Preferably, the persimmon extract comprises persimmon tannin, wherein the content of the persimmon tannin is 15-18%.

Preferably, the horseradish extract comprises horseradish peroxidase, wherein the content of the horseradish peroxidase is 7-10%.

Preferably, the zinc ricinoleate is in liquid state, and the content of the effective component is 30-35%.

Preferably, the emulsifier is a mixture of Tween-80 and Span-80, and the mixture ratio of the Tween-80 to the Span-80 is Tween-80: span-80 ═ 6: 4.

preferably, the stabilizer is a mixture of acetate buffer, lysozyme and glycerol, and the pH regulator is citric acid or lactic acid or a combination thereof.

A method for preparing the composite biological deodorization spray for the pets comprises the following steps:

step 1, taking a certain amount of purified water, then sequentially adding yucca extract, green tea extract, persimmon extract and zinc ricinoleate according to the proportion, and stirring to dissolve the mixture in sequence to obtain solution A;

step 2, weighing an emulsifier, sequentially adding the oregano oil extract, horseradish peroxidase and a stabilizer, and stirring to dissolve the mixture to obtain a solution B;

step 3, pouring the solution B and the solution A into a high-shear homogeneous vacuum mixing and stirring emulsifying machine for stirring at 3000-4000 rpm for 15-20min to obtain a light yellow transparent liquid with uniform color;

and 4, adding a pH value regulator to regulate the mixed solution of the solution A and the solution B to a pH value of 6.5-7.5, and finally adding purified water to a specified volume to obtain the deodorizing spray.

Compared with the prior art, the invention has the following beneficial effects:

1. the deodorizing spray has good deodorizing property on odor molecules of various sources such as pet halitosis, body odor, excrement odor and the like. The main components of the odor are ammonia gas, hydrogen sulfide, methyl mercaptan, trimethylamine and the like, and various odor molecules can be effectively removed by various deodorization components contained in the deodorization spray agent through various different action mechanisms such as catching, adsorption, decomposition and the like, so that the deodorization efficiency is obviously improved.

2. The deodorizing spray has the sterilization characteristic and can radically remove odor. The phenolic compounds in various plant extracts contained in the deodorizing spray have obvious antibacterial, bactericidal, antioxidant and antiseptic effects.

3. The deodorizing spray of the invention has rapid reaction and high deodorizing efficiency. The deodorizing spray can rapidly deodorize, kill bacteria and eliminate viruses, can be used for treating symptoms and root causes, atomize the contained effective components by a spraying method, has the radius of liquid drops which are dispersed in space and is less than or equal to 0.04mm, has large specific surface area and large surface energy, and has the average surface energy of about dozens of kilocalories per mole, the surface energy of the solution can effectively adsorb peculiar smell molecules in the air, simultaneously can change the three-dimensional configuration of the adsorbed peculiar smell molecules, weaken chemical bonds in the peculiar smell molecules, increase the instability of the peculiar smell molecules, easily perform chemical reaction with other molecules and an acid buffer solution in plant liquid, finally generate tasteless and nontoxic substances, eliminate the stink of pet excrement garbage and the like within 5 minutes, inhibit mosquitoes and flies from laying eggs, purify the environment and reduce diseases.

4. The deodorizing spray has good deodorizing effect and long duration. The deodorizing spray can radically inhibit the generation of odor, has long deodorizing duration and can maintain the deodorizing effect for more than 72 hours.

5. The deodorizing spray of the invention is a safe and environment-friendly green product. The deodorizing spray disclosed by the invention is prepared from nontoxic, harmless and non-irritant raw materials, does not contain components such as alcohol and essence, can be safely used for oral cavity deodorization, body deodorization, excrement deodorization and the like, has no harm to people, pets and the environment, and can be safely and safely used for disinfection, sterilization and deodorization in the oral cavity environment of pets, excrement, body peculiar smell, cages, bowls, pet hair and the like. Meanwhile, the product has no corrosiveness and no oxidability, can reduce the harm of disinfectants, chemical agents and the like to the health of pets, and creates an ecological, non-toxic and fragrant growth environment for human beings and animals.

6. The deodorizing spray has strong stability. The emulsifier and the stabilizer contained in the deodorizing spray can keep all components in the formula in a dissolved state, and the deodorizing spray does not have the properties of layering, precipitation, turbidity and the like, so that the product stability is stronger.

7. The deodorizing spray product of the invention is convenient to use. The sterilization component of the deodorizing spray is mainly a plant extraction component, so the deodorizing spray can be directly sprayed for use without dilution, and is convenient for pet owners and pet doctors to use.

8. The deodorizing spray has low cost. The deodorizing spray is a combination of plant type deodorizing spray and biological enzyme type deodorizing spray, and has the advantages of simple preparation method, low cost and suitability for market popularization.

9. The deodorizing spray has wide application range. The deodorizing spray can eliminate various odor in the air, can also eliminate odor generated in the oral cavity and the body of a pet, and can also inhibit bacteria and sterilize, so that the deodorizing spray has wider application range.

Detailed Description

The technical solutions of the present invention will be described clearly and completely with reference to specific embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The compound biological deodorization spray for pets is a novel compound biological deodorization spray which is stable, synchronous and efficient in sterilization and deodorization, free of irritation, convenient to use, safe and green and is prepared by scientifically composing and taking yucca extract, green tea extract, oregano oil extract, persimmon extract, horseradish extract and zinc ricinoleate as main active ingredients and adding an emulsifier, a stabilizer, a pH value regulator and the like as auxiliary ingredients. The emulsifier is a mixture of Tween-80 and Span-80, and the mixture ratio of the Tween-80 to the Span-80 is as follows: span-80 ═ 6: 4, the stabilizing agent is a mixture of acetate buffer, lysozyme and glycerol, and the pH value regulator is citric acid or lactic acid or a combination thereof.

The yucca extract comprises saponin, polysaccharide and polyphenol, wherein the content of saponin is 10-12%, the content of polysaccharide is 10-12%, the content of polyphenol is 15-17%, the B50 value is less than 4mg, the B50 value is used for measuring the ammonia binding capacity of the yucca extract of genus yucca, and the quantity of yucca extract of genus yucca required for binding 50% of ammonia is determined, wherein the lower the value is, the better the value is. The yucca saponin in the yucca extract can effectively combine odor molecules such as ammonia gas and hydrogen sulfide generated by pet excrement, and is a urease inhibitor which can effectively inhibit the generation of ammonia gas from the source; the polysaccharide component of yucca extract has very strong affinity with ammonia gas and is easily combined with ammonia gas, thereby inhibiting the harmful effect of ammonia and converting it into harmless nitride; the polyphenol in Yucca Schidigera extract has strong antioxidant and free radical scavenging ability.

The green tea extract comprises tea polyphenols, wherein the content of tea polyphenols is 20-25%, and the tea polyphenols in the green tea extract has an odor component of methyl mercaptan (CH) generated by oral microorganisms₃SH) has good cleaning effect. The oregano oil extract comprises thymol, polyphenol and carvacrol, wherein the content of thymol is 10-11%, the content of polyphenol is 7-8%, and the content of carvacrol is 8-10%. The persimmon extract comprises persimmon tannin, wherein the content of the persimmon tannin is 15-18%, and the persimmon tannin in the persimmon extract is a polyphenol substance, has good antioxidant activity and strong deodorization performance, and has strong deodorization performance on trimethylamine. And thymol, polyphenol, carvacrol and the like contained in the oregano oil extract, and polyphenol structure in persimmon tannin can react with protein, polysaccharide, alkaloid and the like to change the physical and chemical behaviors, so the method has unique advantages in the aspect of microbial inhibition.

The horseradish extract comprises horseradish peroxidase, wherein the content of the horseradish peroxidase is 7-10%, horseradish catalase in the horseradish extract has a removal effect on 12 odoriferous substances, the horseradish catalase can completely remove phenolic odoriferous substances, the deodorization effect can be maintained for more than 72h, and the horseradish catalase also has a good removal effect on volatile fatty acids. The zinc ricinoleate is in liquid state, the content of active ingredients is 30-35%, the zinc ricinoleate has the greatest advantages of low absorption toxicity through skin, mouth and respiratory tract, safe and reliable use, can effectively eliminate main odor components of ammonia gas, hydrogen sulfide and organic sulfide, and obviously reduces odor intensity. The preparation method of the deodorizing spray comprises the following steps:

The deodorizing spray prepared by the preparation method has the advantages of good deodorizing effect, long duration and stable properties. In order to further understand the present invention, the following will specifically describe the composite biological deodorant spray for pets and the preparation method thereof in combination with the examples.

Example 1

A composite biological deodorization spray for pets comprises the following components in percentage by mass:

the preparation method comprises the following steps:

(1) firstly, taking a certain amount of purified water, sequentially adding yucca extract, green tea extract, persimmon extract and zinc ricinoleate, adding, and stirring to sequentially dissolve to obtain solution A for later use;

(2) then weighing the emulsifier, and sequentially adding the following raw materials: stirring and dissolving the oregano oil extract, horseradish peroxidase and a stabilizer in sequence to obtain a solution B for later use;

(3) pouring the solution B and the solution A into a high-shear homogenizing vacuum mixing stirring emulsifying machine, and stirring at 3000 r/min for 20min to obtain a light yellow transparent liquid with uniform color;

(4) adding a pH value regulator to regulate the mixed solution of the solution A and the solution B to a pH value of 6.5-7.5, and finally adding purified water to 1L to obtain the deodorizing spray.

Example 2

the preparation method comprises the following steps:

(3) pouring the solution B and the solution A into a high-shear homogenizing vacuum mixing and stirring emulsifying machine, and stirring at 4000 rpm for 15min to obtain a light yellow transparent liquid with uniform color;

Example 3

the preparation method comprises the following steps:

(3) pouring the solution B and the solution A into a high-shear homogenizing vacuum mixing stirring emulsifying machine, and stirring at 3500 rpm for 20min to obtain a light yellow transparent liquid with uniform color;

Example 4

the preparation method comprises the following steps:

(3) pouring the solution B and the solution A into a high-shear homogenizing vacuum mixing stirring emulsifying machine, and stirring at 4000 rpm for 20min to obtain a light yellow transparent liquid with uniform color;

Example 5

the preparation method comprises the following steps:

(3) pouring the solution B and the solution A into a high-shear homogenizing vacuum mixing stirring emulsifying machine, and stirring at 3300 r/min for 20min to obtain a light yellow transparent liquid with uniform color;

Example 6

the preparation method comprises the following steps:

(3) pouring the solution B and the solution A into a high-shear homogenizing vacuum mixing stirring emulsifying machine, and stirring at 3800 rpm for 17min to obtain a light yellow transparent liquid with uniform color;

Example 7

the preparation method comprises the following steps:

(3) pouring the solution B and the solution A into a high-shear homogenizing vacuum mixing stirring emulsifying machine, and stirring at 3500 rpm for 18min to obtain a light yellow transparent liquid with uniform color;

Example 8

the preparation method comprises the following steps:

(3) pouring the solution B and the solution A into a high-shear homogenizing vacuum mixing and stirring emulsifying machine, and stirring at 3000 r/min for 15min to obtain a light yellow transparent liquid with uniform color;

Test example 1

The deodorizing spray was subjected to an appearance inspection test to examine whether the deodorizing spray was uniform in appearance, and whether delamination or precipitation occurred.

The deodorizing sprays of examples 1 to 8 were fully shaken in an original packaging container, and then not less than 500ml of stock solution was taken, 100ml of the deodorizing spray stock solution was taken from the stock solution and placed in a 100ml measuring cylinder with a plug, and after being placed for 24 hours at 15 to 35 ℃, the appearance of the deodorizing spray stock solution was observed according to the liquid appearance inspection standard of the pharmacopoeia of the people's republic of China.

The results of the appearance inspection test of the deodorizing sprays are shown in table 1, and it can be seen from table 1 that the deodorizing sprays of examples 1 to 8 are uniform in appearance and have no delamination and precipitation, and all meet the standards of the appearance inspection of the liquid.

Table 1 results of visual inspection of formulations of different examples

Test example 2

And (3) pH value investigation test of the deodorizing spray, and detecting the pH value of the deodorizing spray stock solution.

The deodorizing sprays of examples 1 to 8 were shaken well in the original packaging container, and then a certain amount of deodorizing spray stock solution was taken, and the pH value of the deodorizing spray stock solution was tested by a pH meter at room temperature (25 ℃ ± 2 ℃) to be accurate to 0.1pH unit, and the test results are shown in table 2, and as can be seen from table 2, the pH of the deodorizing spray stock solutions of examples 1 to 8 were all between 6.5 and 7.5, and were all neutral, safe to use, non-corrosive, and safe to be used for spraying in oral cavity, body and environment.

Table 2 pH measurements of different example formulations

Test example 3

The high and low temperature stability test of the deodorizing spray comprises the steps of treating a deodorizing spray sample at high temperature and low temperature, and observing whether the deodorizing spray sample has the phenomena of layering, precipitation and the like.

The test method of the high temperature stability test comprises the steps of pouring the deodorization spray into a 50ml measuring cylinder with a plug to 50ml scales, placing the measuring cylinder with the plug into a constant temperature water bath with the temperature of 60 +/-2 ℃ after the plug is added, taking the measuring cylinder out after the water surface is 20mm higher than the scales and recovering to the room temperature after 6 hours, and observing the appearance, wherein the results are shown in table 3.

Test method of low temperature stability test the sample which has not changed in the high temperature test (if it has changed, another new sample is needed) is placed in a refrigerator of minus 5 +/-2 ℃ for 2h, and then taken out, and after it naturally returns to room temperature, the appearance is observed, and the result is shown in table 3.

As can be seen from table 3, the deodorizing spray samples of examples 1 to 8 were stable, uniform, and non-delaminated under both high temperature and low temperature conditions, and no crystal or precipitate was precipitated.

TABLE 3 high and Low temperature stability Change test results for different example formulations

Test example 4

The deodorizing effect test of the deodorizing spray evaluates the deodorizing effect of each of the deodorizing spray formulas of examples 1 to 8 by the deodorizing effect of several odor molecules common to pets.

The test reagent used in the test for testing the deodorization effect of the deodorization spray is ammonia water, hydrogen sulfide, methyl mercaptan, methyl sulfide and dimethyl disulfide. The test instruments used for the test of the deodorization effect of the deodorization spray agent comprise an ammonia and hydrogen sulfide gas detection tube, an HP5840A gas chromatograph, a 5.5L glass bottle and an injector.

1. Odor instrument analysis method

Ammonia gas, hydrogen sulfide and 3 kinds of organic sulfides with different concentrations are respectively prepared in a 5.5L glass bottle, the deodorization effect is evaluated by absorbing and fixing ammonia gas with boric acid, and the deodorization effect of the deodorization spray on the removal effect of hydrogen sulfide, methyl mercaptan, dimethyl disulfide and dimethyl disulfide is evaluated by headspace gas chromatography. Gas chromatography conditions: column DB-1 with injection port temperature of 170 deg.C, column temperature of 40 deg.C, checker temperature of 250 deg.C, ammonia gas-carrying gas (60ml/min), hydrogen as fuel, and pressure of 1kg/cm²Air combustion supporting, pressure 1.5kg/cm². Weighing 0.01g of sample, dissolving in a headspace bottle with 1ml of water, injecting a certain amount of odor substance in a sealing way, and keeping for 1h to absorb 1ml of headspace air for sample injection. The concentration of each gas before and after 1 hour of action was measured, and the elimination ratio was calculated, and a blank control (purified water) was also used in the experiment.

The deodorizing effect on a single odor molecule is shown in table 4, and it can be seen from table 4 that the deodorizing spray of the present invention can effectively and rapidly remove odors such as ammonia, hydrogen sulfide, methyl mercaptan, dimethyl sulfide, dimethyl disulfide, etc., and the elimination rate of these 5 single odor molecules can reach 85% or more.

Table 4 individual deodorizing effects of the different example formulations on 5 odors

2. Olfaction evaluation method

The mixed gas of ammonia gas, hydrogen sulfide, methyl mercaptan, methyl sulfide and dimethyl disulfide is prepared in a bottle, a six-grade odor intensity representation method (shown in table 5) is adopted, 5 trained sniffers are employed to measure the odor intensity before and after elimination, the average value of the odor intensity is calculated, and the elimination rate is calculated at the same time.

The results of the comparison experiments using purified water as the reference are shown in table 6, and it can be seen from table 6 that the deodorizing effect of the deodorizing spray of examples 1 to 8 of the present invention is excellent, and the ratio of the elimination rate of the composite odor molecules can be up to 85% or more.

TABLE 5 odor degree table according to smell sense

Table 6 deodorant effect of different example formulations on 5 mixed odors table

Test example 5

The safety test of the deodorizing spray evaluates the safety of the deodorizing spray formulas in the examples 1 to 8 through an acute toxicity test, a subacute toxicity test, a skin toxicity test, an animal eye combined membrane irritation test and a human skin local irritation test.

1. Test material and test object

Test materials: deodorizing spray suspension containing cod liver oil (containing 1500U vitamin A and 500U vitamin D/g) for acute toxicity test; mixed feed containing 0.5% of deodorizing spray for subacute test; glycerin suspension of 25% deodorant spray for skin toxicity test; cod liver oil suspension of 5% deodorant spray for eye conjunctival irritation test; the human skin irritation test uses a 15% deodorant spray petrolatum mixture.

Test subjects: the acute toxicity test uses 40 wistar female rats, the subacute toxicity test uses 40 wistar rats (male half and female half), the skin toxicity and irritation test uses 15 female guinea pigs, the eye conjunctiva irritation test uses 10 guinea pigs and rabbits, and the human skin local irritation test uses 5 healthy men and women in Zhuang years.

2. Acute toxicity test

Animals were kept alive following intragastric administration of 6.8g/kg and 8g/kg of the deodorant spray of example 1 in preliminary experiments. The test dosage is set to be 9.0g/kg of deodorization spray, namely 5-8 mL of cod liver oil suspension. Control animals were given equal amounts of cod liver oil. The test is performed by fasting for 1 night and filling medicine on day 2, then the general condition and survival condition of the animals are observed, the animals are killed by cutting off the heads on day 8, the lung, the liver and the kidney are taken out, and the tissue morphology change is observed under an optical microscope. Immediately after killing animals, the lung, heart, liver, kidney and blood (separated serum) were placed in 25ml beakers, respectively, 5ml nitric acid and 2ml perchloric acid were added, and after nitration, the zinc content was measured by atomic absorption spectrometry.

During the test period, 20 rats each with 9.0g/kg body weight in the gavage test group and the control group died, so that the LD of the rats can be confirmed₅₀The value (median lethal dose) is far above 9.0g/kg, and due to limited gastric volume, LD is difficult to obtain₅₀The specific numerical value of (1).

The animals in the experimental group and the control group breathe faster than the original animals after the gavage, do not love the activity, last for about 1 day, and the appetite is reduced to some extent in the period. After one week, the body weight of the experimental group increased from 178.4g to 216.3g before the experiment, and the body weight increased 21.2% averagely; the control group increased from 182.6g to 217.8g before the test, with an average increase of 19.3%, and there was no significant difference between the statistically treated groups, as detailed in table 7.

Observing the liver cells, sinusoids, sinus walls, cortex and medulla of kidney by tissue morphology; no specific pathological changes are found on the alveolar wall of the lung.

After one week of gavage, the zinc contents of the major organs and serum of the animals are almost similar, and no significant difference is observed in statistical treatment, which indicates that no zinc is accumulated, and the details are shown in Table 8.

The acute toxicity test results of examples 2 to 8 are the same as those of example 1, and are not repeated here.

TABLE 7 acute toxicity test animal body weight Change (g)

n＝20

TABLE 8 acute toxicity test Zinc content (ug/g) in organs and serum

n is 10(m + -SD) units umol/L

3. Subacute toxicity test

The test animals were randomly divided into two groups of experimental and control animals, and were housed in 4 cages. The test animals were fed with the normal feed containing 0.5% of the deodorizing spray of example 1, and the control was fed with the normal feed, with water ad libitum. Weighing once a month, performing abdominal incision with 2.2% sodium pentobarbital by intraperitoneal injection anesthesia (about 1m 1) after 3 months, performing abdominal active and venous bloodletting, taking lung, liver and kidney tissues, performing routine pathological examination on one part, observing tissue morphological change, and measuring zinc content in the tissue on the other part.

The animals in both groups survived all the time during the 90d period and gained weight, and the results are shown in Table 9. As can be seen from Table 9, the body weight of the experimental group increased 96.9% in 3 months, the body weight of the control group increased 87.2%, and there was no significant difference between the two groups.

Observing the liver cells, sinusoids, sinus walls, cortex and medulla of kidney by tissue morphology; no specific pathological changes are found on the alveolar wall of the lung. The result of histomorphosis observation shows that the pathological changes of lung, liver and kidney tissues of the experimental group and the control group have no obvious difference.

The results of measuring the zinc content in the major organs of the subacute toxicity test animals are shown in table 10, and it is understood from table 10 that the zinc content in the major organs of the animals is substantially similar, and there is no significant difference in statistical treatment, indicating that there is no zinc accumulation.

The subacute toxicity test results of examples 2 to 8 are the same as those of example 1, and are not repeated herein.

TABLE 9 subacute toxicity test animal Change

n＝40(M±SD)P＞0.5

TABLE 10 Zinc content (ug/g) in major organs of test animals

n＝10(M±SD)P＞0.2

4. Skin toxicity test

Before the test, the guinea pigs are cut short and long on the back, coated with depilatory (prepared from 10% sodium sulfide and Arabic gum), shaved after 5-7 min, washed and observed for 24h, and the guinea pigs without obvious redness and swelling and skin injury are selected as test objects. The midline of the back of guinea pig is used as boundary, and the two sides are respectively test zone and control zone, each of which is about 6cm²The test area was coated with 25% of the deodorant spray glycerin of the formulation of example 1 (1:1 aqueous solution) to cover the skin and short hairs 1 time a day for 5 consecutive days. And (4) coating a 50% glycerol aqueous solution as a control, and observing whether the epidermis has red and swollen spots and the like day by day. And 6, killing the broken ends and the living ends, respectively adopting conventional staining of the skin of two areas, and observing the morphological change of the tissues under an optical microscope.

The results of 5d observation after the deodorant spray of the formula of example 1 was applied showed that no obvious irritation symptoms and red, swollen, blotchy and other lesions were observed on the skin surface in the test area and the control area. Histological morphology observation under a light lens shows that the skins of two groups of animals belong to normal skins.

The results of the skin toxicity tests of examples 2 to 8 are the same as those of example 1, and are not repeated here.

5. Animal eye binding membrane irritation test

5% of the deodorizing spray cod liver oil suspension 2 drops according to the formula of example 1 is dripped into the left eye of a rabbit or a guinea pig, and only 2 drops of cod liver oil are dripped into the right eye, and the conjunctiva of the eye is observed for the existence of hyperemia, lacrimation and the like immediately after dripping, 10min, 30min, l h and 4h respectively.

The result of the animal eye combined membrane stimulation test of the deodorizing spray prepared in the example 1 shows that 2 medicinal eyes and 1 control eye in rabbits have mild mucosal congestion within 30min after the application of the deodorizing spray, and then the deodorizing spray is recovered to be normal, and no obvious stimulation reaction is found after the guinea pig eyes are applied.

The results of the animal eye binding membrane stimulation tests of examples 2 to 8 are substantially the same as those of the animal eye binding membrane stimulation test of example 1, and the detailed description thereof is omitted.

6. Local irritation test of human skin

A petrolatum paste containing 15% of the deodorizing spray of the formulation of example 1 was prepared, applied to the skin of one forearm of the subject, covered with a gauze with a paper pad, and wrapped 1 time a day for 5 days, and the control was coated with petrolatum paste only, and the irritation to the skin was observed.

The result of the local irritation test of the human skin of the deodorant spray prepared according to the formula in the example 1 shows that 5 primary reactions of redness, itching, rash, and the like occur on 3 persons in 10 subjects in the application process, wherein no obvious difference is found when 3 times are performed on one side of the test and 2 times are performed on the other side of the control.

The results of the human skin local stimulation tests of examples 2 to 8 are substantially the same as those of the human skin local stimulation test of example 1, and are not repeated herein.

According to the results, the selected formula products in the examples 1 to 8 are extremely high in safety as an external pet product, and the external pet product is almost non-toxic to eyes, skin, oral cavity and the like of a pet, is extremely high in safety and can be licked safely by the pet through tests such as an acute toxicity test, a sub-acute toxicity test, a skin toxicity test, an animal eye binding membrane irritation test, a human skin local irritation test and the like.

Test example 6

The deodorizing spray is used for testing the on-site deodorizing effect of a dog factory. In the test, a large dog factory in Shandong is selected, and in winter with poor ventilation conditions, the deodorizing effect of the deodorizing spray formulas in the examples 1 to 8 on the dog factory is verified through instrument detection.

Selecting 10 kennels in the large-scale dog factory, performing on-site ammonia detection on the kennels by using a portable ammonia gas detector before deodorization and after deodorization for 5 minutes respectively, specifically, closing doors and windows 2 hours before the detection, performing ammonia gas value detection on 3 different distribution points of each kennels at a height of 0.9 meter from the ground, and averaging.

The test results are shown in table 11, and it can be seen from table 11 that the deodorizing sprays of the formulas of examples 1 to 8 have good deodorizing effect on the site of dog factories, can rapidly eliminate ammonia gas after 5 minutes of action, and the percentage of ammonia reduction reaches more than 75%, so that the air in the dog house is effectively purified, the respiratory system diseases are reduced, and the deodorizing effect of the deodorizing spray of the invention is superior to that of the similar deodorizing sprays in the market.

TABLE 11 results of the deodorization effectiveness test of different formulations of examples on dog factories

Test example 7

The deodorization spray is used for a test for on-site sterilization and disinfection of a dog factory. In the experiment, a large dog factory in Shandong is selected, and the killing effect of the deodorizing spray formulas in examples 1 to 8 on natural bacteria on the surfaces of the dog factory is verified.

1. Test reagents and materials

Phosphate buffered saline (PBS, 0.03mol/L, pH 7.2), diluent (PBS solution containing 0.1% Tween 80), sterile cotton swab, format plate (made of stainless steel material with 5.0cm × 5.0cm of blank space left in the center as sampling site), and nutrient agar medium.

2. Test method

(1) Randomly taking the surface of the wall, and calibrating 3 blocks by using a specification board to obtain areas of 25cm respectively²The block of (1) is used for sampling before disinfection, sampling after chemical disinfection and sampling by deodorization spray disinfection.

(2) Spraying purified water on the sampling area, wetting a sterile cotton swab in a test tube containing 5ml of diluent, smearing and sampling a block, and reciprocating 8 times in the horizontal direction and the vertical direction. After sampling, the sampling end of the cotton swab is cut into the original diluent test tube in a sterile operation mode, and is shaken for 20s or rapped for 80 times, and after appropriate dilution, the cotton swab is used as a negative control group sample.

(3) Diluting the chemical disinfectant according to the instruction amount, spraying the diluted chemical disinfectant to a sampling area for disinfection, wetting a sterile cotton swab in a test tube containing 5ml of diluent, smearing and sampling a block, and reciprocating 8 times horizontally and vertically. After sampling, the sampling end of the cotton swab is cut into the original diluent test tube in a sterile operation mode, and is shaken for 20s or rapped for 80 times, and after appropriate dilution, the cotton swab is used as a positive control group sample.

(4) Directly spraying the deodorizing spray to a sampling area for deodorizing and disinfecting, wetting a sterile cotton swab in a test tube containing 5ml of diluent, smearing and sampling a deodorizing and disinfecting area, and reciprocating 8 times horizontally and vertically. After sampling, the sampling end of the cotton swab is cut into the original release liquid test tube in a sterile operation mode, and the electric mixer vibrates for 20s or raps for 80 times to be used as a sample of the deodorization and disinfection group.

(5) And (3) sucking 1.0ml of the positive control group sample, the negative control group sample and the deodorization and disinfection group sample in each part, inoculating the plates by an agar pouring method, inoculating 2 plates in each sample, culturing for 48 hours in a 37 ℃ incubator, and observing the final result.

(6) Calculating kill logarithm value

The experiment was repeated 3 times, and the number of colonies (cfu) for each group was calculated and converted to a log value (N), and then a log kill value was calculated as follows:

log Kill (KL) is log of average viable bacteria concentration of control group (No) -log of viable bacteria concentration of experimental group (Nx)

The test is repeated for 3 times, more bacteria should grow in the positive control group, more bacteria should grow in the negative control group, the average killing log value of the disinfection sample is more than or equal to 1, and the disinfection is qualified.

3. Test results

The test results of the tests of the deodorant spray on the field sterilization and disinfection test of the dog factory are shown in table 12, and it can be seen from table 12 that the deodorant sprays of the formulas of examples 1 to 8 have good effects on the field sterilization and disinfection of the dog factory, and the average killing logarithm value of each group is more than or equal to 1, and is similar to the effect of a positive control group (chemical disinfectant). Therefore, the deodorizing spray prepared according to each of the formulas of the embodiments 1 to 8 can deodorize and efficiently sterilize, so that the odor problem can be solved essentially and fundamentally.

TABLE 12 results of the on-site sterilization and disinfection test of different example formulations in dog factories

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims

1. A preparation method of a composite biological deodorization spray for pets is characterized by comprising the following steps:

step 4, adding a pH value regulator to regulate the mixed solution of the solution A and the solution B to a pH value of 6.5-7.5, and finally adding purified water to a specified volume to obtain the deodorizing spray;

the components comprise, by mass, 5-8% of yucca extract, 4-5% of green tea extract, 6-8% of oregano oil extract, 4-5% of persimmon extract, 4-5% of horseradish extract, 3-4% of zinc ricinoleate, 5-6% of emulsifier, 1-2% of stabilizer, 1-2% of pH value regulator and the balance of purified water, wherein,

the yucca extract comprises saponin, polysaccharide and polyphenol, wherein the content of saponin is 10-12%, the content of polysaccharide is 10-12%, and the content of polyphenol is 15-17%;

the green tea extract comprises tea polyphenols, wherein the content of the tea polyphenols is 20-25%;

the oregano oil extract comprises thymol, polyphenol and carvacrol, wherein the content of thymol is 10-11%, the content of polyphenol is 7-8%, and the content of carvacrol is 8-10%;

the persimmon extract comprises persimmon tannin, wherein the content of the persimmon tannin is 15-18%;

the horseradish extract comprises horseradish peroxidase, wherein the content of the horseradish peroxidase is 7-10%; the zinc ricinoleate is in a liquid state, and the content of the effective component is 30-35%;

the emulsifier is a mixture of Tween-80 and Span-80, and the mixture ratio of the Tween-80 to the Span-80 is as follows: span-80 ═ 6: 4;

the stabilizer is a mixture of acetate buffer solution, lysozyme and glycerol, and the pH value regulator is citric acid or lactic acid or a combination thereof.