CN115251092B - Environment-friendly disinfectant capable of rapidly inactivating bacteria and viruses and preparation method thereof - Google Patents
Environment-friendly disinfectant capable of rapidly inactivating bacteria and viruses and preparation method thereof Download PDFInfo
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- CN115251092B CN115251092B CN202211065164.3A CN202211065164A CN115251092B CN 115251092 B CN115251092 B CN 115251092B CN 202211065164 A CN202211065164 A CN 202211065164A CN 115251092 B CN115251092 B CN 115251092B
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Classifications
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- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
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- B01D53/007—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols by irradiation
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
- B01D53/34—Chemical or biological purification of waste gases
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- B01D2255/802—Photocatalytic
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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Abstract
The invention provides an environment-friendly disinfectant capable of rapidly inactivating bacterial viruses and a preparation method thereof, wherein the disinfectant is mainly prepared from the following raw materials: 60-80 parts of single-chain quaternary ammonium salt, 10-15 parts of acorn extract, 10-15 parts of polygonum orientale extract, 2-8 parts of Galium aparine extract, 2-8 parts of photocatalyst, 5-10 parts of phosphoric acid, 2-8 parts of radix puerariae powder, 1-5 parts of ganoderma lucidum powder, 1-5 parts of trehalose and 1-5 parts of maltose. The disinfectant prepared by the invention has the advantages of strong sterilization capability, no toxicity, no stimulation and no corrosion, low cost, simple preparation steps and capability of meeting the use requirements of most people.
Description
Technical Field
The invention relates to the technical field of disinfectants, in particular to an environment-friendly disinfectant capable of rapidly inactivating bacterial viruses and a preparation method thereof.
Background
The disinfectant is used for killing pathogenic microorganisms on a transmission medium to achieve the harmless requirement, and the pathogenic microorganisms are killed outside human bodies to cut off the transmission path of infectious diseases so as to achieve the aim of controlling the infectious diseases.
The most difficult to solve infectious diseases at present is the new coronavirus epidemic situation caused by the new coronavirus, the new coronavirus is popular in the world at present, the new coronavirus is mainly transmitted through respiratory tract droplets and closely contacts and transmitted, and the new coronavirus has stronger viability in the external environment through research, so that in order to protect the new coronavirus, the current people can spray the disinfectant when going home or taking express, so as to kill the virus, but the existing disinfectant has a certain difference in the killing effect on the new coronavirus, and the risk of being infected by the virus is increased.
In view of this, the present invention has been made.
Disclosure of Invention
The first object of the invention is to provide an environment-friendly disinfectant capable of rapidly inactivating bacteria and viruses, which has good sterilizing effect, can kill most of bacteria and ensures the safety of people.
The second object of the invention is to provide a preparation method of the disinfectant, which is simple and quick, and can prepare the disinfectant which meets national regulations, has good sterilization effect and is safer.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
the invention provides an environment-friendly disinfectant capable of rapidly inactivating bacterial viruses, which is mainly prepared from the following raw materials:
60-80 parts of single-chain quaternary ammonium salt, 10-15 parts of acorn extract, 10-15 parts of polygonum orientale extract, 2-8 parts of Galium aparine extract, 2-8 parts of photocatalyst, 5-10 parts of phosphoric acid, 2-8 parts of radix puerariae powder, 1-5 parts of ganoderma lucidum powder, 1-5 parts of trehalose and 1-5 parts of maltose.
Further, the main raw materials for preparing the disinfectant are selected as follows: 65-78 parts of single-chain quaternary ammonium salt, 12-14 parts of acorn extract, 12-14 parts of polygonum perfoliatum extract, 3-7 parts of Galium aparine extract, 3-7 parts of photocatalyst, 6-9 parts of phosphoric acid, 4-7 parts of radix puerariae powder, 2-4 parts of ganoderma lucidum powder, 2-4 parts of trehalose and 2-4 parts of maltose.
Further, the main raw materials for preparing the disinfectant are as follows: 70 parts of single-chain quaternary ammonium salt, 13 parts of acorn extract, 13 parts of polygonum multiflorum extract, 5 parts of Galium verum extract, 5 parts of photocatalyst, 7 parts of phosphoric acid, 5 parts of kudzuvine root powder, 3 parts of lucid ganoderma powder, 3 parts of trehalose and 3 parts of maltose.
Preferably, the single-chain quaternary ammonium salt is one or a combination of two of dodecyl dimethyl benzyl ammonium chloride and dodecyl dimethyl phenethyl ammonium bromide.
Preferably, the photocatalyst is a nano silica photocatalyst.
Preferably, the nano silicon dioxide photocatalyst contains rare earth elements and boron elements, wherein the rare earth elements are at least one of lanthanum, cerium, praseodymium or neodymium.
In the invention, the raw materials of the disinfectant are selected to be the single-chain quaternary ammonium salt, and the single-chain quaternary ammonium salt can be hydrolyzed in water to generate positive charges, is adsorbed on the surfaces of microorganisms to form ion groups and permeates the quasi-support and protein layers of bacterial plasma membranes, so that the permeability of the plasma membranes is changed, the sterilizing effect is achieved, the disinfectant has no residue, is beneficial to treatment, has little skin irritation to human bodies, does not have toxicity accumulation after long-term use, and has sterilizing and deodorizing effects.
In order to achieve a better sterilization effect, phosphoric acid is added into the raw materials, and phosphate can be formed after the phosphoric acid is esterified, so that the main component of the phosphate is the same as the main component of lipid of cell membranes of bacteria, the phosphate has better affinity with the bacteria, and meanwhile, the phosphate is matched with single-chain quaternary ammonium salt for use, so that the phosphate can better permeate into the bacteria for sterilization, and the sterilization effect is improved.
Meanwhile, the invention also adopts acorn extract and polygonum extract, and the extracts belong to natural plant extracts, so that the natural plant extract has rich content in nature and low cost, and simultaneously contains flavonoids, phenols, essential oil and the like, thereby having the function of bacteriostasis, resisting oxidation and preventing deterioration.
Through researches, it can be known that viruses generally exist in the environment and can be attached to clothes, so that in order to clean some viruses on the clothes, a Galium extract is selected in the raw materials, and can be quickly polymerized, flocculated and then shed with various impurities and peculiar smell molecules of the clothes, so that the comprehensive penetration of a disinfectant is facilitated, the thoroughness of the disinfection is improved, and meanwhile, the effects of sterilization and antibiosis are also achieved.
In order to prevent viruses in the air from infecting the human body, the photocatalyst is added, and the photocatalyst can effectively degrade toxic and harmful gases in the air, such as formaldehyde and the like, so that the air is purified efficiently; meanwhile, the photocatalyst can effectively kill various bacteria, can decompose and harmlessly treat toxins released by the bacteria or fungi, has stable chemical properties, does not cause secondary pollution and cannot pollute the environment, and has better sterilization capability when doped with rare earth elements and boron elements.
Although the disinfectant can play a role in disinfection and sterilization and possibly cause certain harm to human bodies, the disinfectant is proved to be nontoxic, non-irritating and non-corrosive by a third-party experiment, in order to prevent the harm, the ganoderma lucidum powder and the kudzuvine root powder are added into the raw materials, and the toxicity of the disinfectant can be reduced by adding the two raw materials, so that the product is more green and natural, and no pollution residue exists on the environment.
Finally, in order to ensure the shelf life of the disinfectant prepared by the invention, maltose and trehalose are selected, which have certain stability, so that the shelf life of the product can be prolonged, and the toxicity of the disinfectant is fully reduced after a small amount of the disinfectant is added, so that the neutralization effect is achieved.
In a word, for the disinfectant disclosed by the invention, the compatibility of the components ensures a good sterilization effect and also fully reduces the toxicity of the disinfectant; meanwhile, experiments show that the novel coronavirus has good inactivation effect on the novel coronavirus. Besides, the sterilizing agent has good inactivating effect on various common viruses in daily life, and particularly, the sterilizing agent fully considers the toxicity of the sterilizing agent while improving the sterilizing effect, and simultaneously considers low toxicity and high sterilizing effect. Especially, the combination of the three extracts increases the permeability while sterilizing, and the dosage is controlled, because if the dosage is too large, the sterilization medium is possibly damaged to some extent, and the disinfectant cannot play a role.
The invention also provides a preparation method of the disinfectant, which comprises the following steps:
mixing single-chain quaternary ammonium salt with phosphoric acid, photocatalyst, acorn extract and Polygonum tinctorium extract, stirring, adding Ganoderma powder, radix Puerariae powder, trehalose, and maltose, heating and stirring to obtain the final product.
Preferably, the phosphoric acid is subjected to an esterification reaction prior to mixing.
Preferably, the stirring speed is 500-600rpm;
preferably, the speed of agitation is 550rpm.
Preferably, the temperature is raised to 50-70 ℃;
preferably, the temperature is raised to 60 ℃.
Preferably, the extraction method of the acorn extract comprises the following steps:
drying, grinding, leaching, concentrating, extracting, degreasing, extracting, filtering and drying acorn to obtain acorn extract;
preferably, the temperature of the drying is 85-95 ℃, and the extraction is selected from one or a combination of two of carbon tetrachloride and ethylene. The temperature of drying is selected to be 85-95 ℃ because the temperature is too low, the rubber is possibly not thoroughly dried, the rubber is softened according to the old water, the temperature is too high, the rubber is not beneficial to the subsequent treatment, carbon tetrachloride and ethylene are selected as extracting agents, the carbon tetrachloride and ethylene are quite stable, the reaction is not easy to occur, and the rubber extract with good effect can be perfectly extracted. The method for preparing the acorn extract provided by the invention is simple to operate, can reduce the cost, can recycle and reuse the used solvent, and is easy to produce.
Preferably, the preparation method of the cortex polygoni orientalis extract comprises the following steps:
performing osmotic extraction on the cortex polygoni multiflori, concentrating, extracting, recovering under reduced pressure, and eluting by adopting silica gel column chromatography to obtain the cortex polygoni multiflori extract;
preferably, the pressure is reduced to 40-48 ℃ and 1-1.5MPa, and the mesh number of the silica gel column adopted by the chromatographic elution method is selected to be 250-300 mesh. Firstly, the invention selects to cool and decompress in order to ensure that the extract of the Polygonum tinctorium linn is in a stable and non-decomposed state when being recovered, and is convenient to recover, and adopts silica gel column chromatography elution to effectively separate and extract the extract of the Polygonum tinctorium linn, so that the extract of the Polygonum tinctorium linn with good disinfection effect can be obtained, and the mesh number of the silica gel column is selected to be 250-300 mesh, so that the extract of the Polygonum tinctorium linn with finer separation can be separated and treated, the separation effect is better, and the extract of the Polygonum tinctorium linn can be better matched with other raw materials to produce disinfection and sterilization effects.
In a word, the purity of the extract obtained by the specific method can reach more than 99%, the impurities are few, and the harmful components contained in the impurities are few, so that the sterilization effect is more ensured after the disinfectant is prepared subsequently, and the low-toxicity safety of the disinfectant can be further ensured.
Through practice, the application effect of the extract obtained by direct purchase in the prior art is poorer than that of the extract of the invention even under the condition that the purity of the extract is consistent with that of the extract of the acorn prepared by the extraction method of the invention, and the reason is probably that the beneficial components of the acorn extract extracted by the extraction method of the invention are more, the quality is more ensured, and the effect is more remarkable after the extract is compatible with other components.
Preferably, the preparation method of the photocatalyst comprises the following steps:
adding rare earth elements and boron elements into the nano silicon dioxide photocatalyst, adding water, stirring, roasting in a muffle furnace, and crushing to obtain modified nano silicon dioxide;
preferably, the stirring speed is 1100-1200rpm, and the temperature of the muffle furnace is 550-650 ℃. The stirring speed is set to ensure that rare earth elements, boron elements and nano silicon dioxide photocatalyst can be thoroughly mixed, the mixing is incomplete due to the too high or too low rotating speed, the modification is incomplete, and the effect can not reach an ideal state when the modified nano silicon dioxide photocatalyst is mixed with other raw materials for use.
The photocatalyst after modification is more beneficial to the exertion of the sterilization effect, and the stability of the photocatalyst material is also improved.
Compared with the prior art, the invention has at least the following advantages:
(1) The raw materials selected by the invention are nontoxic or low in toxicity, and cannot harm human bodies.
(2) The invention has the advantages of common raw materials, low price, simple preparation method, no need of expensive special equipment, and labor and money saving.
(3) The disinfectant prepared by the invention can disinfect the surface of an object, can permeate into clothes to deeply disinfect, can protect the disinfected skin and avoid hurting human bodies during disinfection.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to examples, but it will be understood by those skilled in the art that the following examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
Example 1
The embodiment 1 provides an environment-friendly disinfectant capable of rapidly inactivating bacterial viruses, which is mainly prepared from the following raw materials:
70g of single-chain quaternary ammonium salt, 13g of acorn extract, 13g of polygonum seed extract, 5g of Galium verum extract, 5g of photocatalyst, 7g of phosphoric acid, 5g of kudzuvine root powder, 3g of lucid ganoderma powder, 3g of trehalose and 3g of maltose.
The embodiment also provides a preparation method of the disinfectant, which comprises the following steps:
1. pretreatment of materials is carried out:
(1) Preparation of acorn extract:
washing acorn, oven drying, grinding into acorn powder, adding diethyl ether solution for leaching, performing water bath reflux to obtain leaching solution, concentrating the leaching solution, adding the leaching solution for extraction and degreasing, concentrating into paste in boiling water solution, adding acetone for dissolving, and continuously extracting and filtering to obtain acorn extract.
In order to obtain purer acorn extract, the extract is selected as carbon tetrachloride and the drying temperature is selected as 90 ℃.
(2) Preparation of the extract of cortex polygoni orientalis:
adding methanol solution into cortex Polygoni Multiflori radix, performing osmotic extraction to obtain extractive solution, concentrating the extractive solution to paste, adding water, extracting with dichloromethane, recovering under reduced pressure, and eluting with silica gel column chromatography to obtain cortex Polygoni Multiflori radix extract.
In order to obtain purer extract of Polygonum tinctorium, the present example selects to decompress to 45 deg.C, 1.3MPa, and the mesh number of silica gel column of chromatography is selected to 280 mesh.
(3) Preparation of photocatalyst:
adding rare earth elements and boron elements into the nano silicon dioxide photocatalyst, adding water, stirring for 50min, standing for 1h, drying, putting the dried powder into a muffle furnace for roasting for 1.5h, standing and cooling to room temperature, and crushing to obtain the modified nano silicon dioxide photocatalyst,
In order to obtain a purer photocatalyst, the stirring speed is 1150rpm, and the temperature of the muffle furnace is 600 ℃.
2. Preparing a disinfectant:
(1) Firstly, esterifying phosphoric acid, then sequentially adding single-chain quaternary ammonium salt, photocatalyst, acorn extract and polygonum extract, stirring at a rotation speed of 500-600rpm, and mixing, wherein in the embodiment, the rotation speed is selected to be 550rpm in order to achieve a good effect;
(2) Pulverizing Ganoderma powder, radix Puerariae powder, trehalose and maltose into 90-120 mesh powder, in this embodiment, pulverizing into 100 mesh powder for good effect;
(3) The crushed powder is screened, mixed with the mixture, then added with water and warmed to 50-70 ℃ and mixed at a rotational speed of 500-600rpm until the mixture is completely mixed, in this example, the rotational speed is selected to be 550rpm and the warming is selected to be 60 ℃ in order to achieve a good effect.
In this embodiment, in order to ensure the effect of the disinfectant, the single-chain quaternary ammonium salt is selected to be a mixture of dodecyl dimethyl benzyl ammonium chloride and dodecyl dimethyl phenethyl ammonium bromide, wherein the molar ratio between the dodecyl dimethyl benzyl ammonium chloride and the dodecyl dimethyl phenethyl ammonium bromide is 1:1.5, and the photocatalyst is a nano silicon dioxide photocatalyst containing praseodymium and boron.
Examples 2 to 6
The specific embodiment corresponds to example 1, except that table 1 below shows:
TABLE 1 variation of parts of raw materials
| Example 2 | Example 3 | Example 4 | Example 5 | Example 6 | |
| Single-chain quaternary ammonium salt/g | 60 | 80 | 65 | 78 | 63 |
| Acorn extract/g | 15 | 10 | 14 | 12 | 12 |
| Polygonum tinctorium extract/g | 10 | 15 | 12 | 14 | 14 |
| Galium head extract/g | 8 | 2 | 7 | 3 | 6 |
| Photocatalyst/g | 2 | 8 | 3 | 7 | 4 |
| Phosphoric acid/g | 10 | 5 | 9 | 6 | 8 |
| Radix Puerariae powder/g | 2 | 8 | 4 | 7 | 6 |
| Glossy ganoderma powder/g | 1 | 5 | 2 | 4 | 2 |
| Trehalose/g | 5 | 1 | 4 | 2 | 5 |
| Maltose/g | 1 | 5 | 2 | 4 | 3 |
Example 7
The specific embodiment corresponds to example 1, with the only difference that the single-chain quaternary ammonium salt is selected as dodecyldimethylbenzyl ammonium chloride.
Example 8
The specific implementation mode is consistent with the embodiment 1, and the only difference is that the photocatalyst is a nano silicon dioxide photocatalyst and is not doped with any trace elements.
Example 9
The specific embodiment is the same as example 1 except that the drying temperature is 85 ℃ and the extract is cyclohexane, the pressure is reduced to 48 ℃ and 1MPa, the number of silica gel columns is 250 mesh, the stirring speed is 1100rpm, and the temperature of the muffle furnace is 650 ℃ when preparing the photocatalyst.
Example 10
The specific embodiment is the same as example 1 except that the drying temperature is selected to be 95 ℃ and the extract is selected to be cyclohexane when preparing acorn extract, the pressure is reduced to be 40 ℃ and 1.5MPa when preparing magnolia officinalis extract, the number of silica gel columns is selected to be 300 meshes, the stirring speed is selected to be 1200rpm when preparing photocatalyst, and the temperature of a muffle furnace is 550 ℃.
Comparative example 1
The specific embodiment is identical to example 1, except that the extract of Polygonum tinctorium is not added.
Comparative example 2
The specific embodiment corresponds to example 1, with the only difference that the acorn extract selected is one sold by the company Ningxia vanilla biotechnology Co.
Comparative example 3
The specific embodiment is the same as in example 1, except that the radix Puerariae powder and the Ganoderma lucidum powder are not added.
Comparative example 4
The specific embodiment corresponds to example 1, with the only difference that no trehalose is added.
Comparative example 5
The specific embodiment corresponds to example 1, with the sole difference that 8g of maltose is used.
Comparative example 6
The specific embodiment corresponds to example 1, with the only difference that no phosphoric acid is added.
Experimental example 1 quantitative test of killing of suspension
Example 1, example 3, example 10 and comparative example 1 and comparative example 6 are selected as sample raw materials, escherichia coli, staphylococcus aureus, candida albicans and pseudomonas aeruginosa are selected as experimental strains, a neutralizing agent is added, the experimental strains are put into a biochemical incubator, the effective time is 0.5 times of the effective time under the lowest using concentration, and the control group is not added with any disinfectant, and the experimental strains are obtained by adding physiological saline.
The experimental results are shown in table 2 below:
table 2 results of killing experiments
As is clear from the above experiments, the disinfectant prepared by the invention can play a role in killing most harmful strains in a short time, and can meet the daily demands of most people, and the disinfectant adopted in the comparative example 1 and the comparative example 6 has greatly reduced sterilization effect because the comparative example 1 does not adopt the Polygonum tinctorium extract, and has a good sterilization synergistic relationship because the three extracts are required to be matched in proper proportion, and when the dosage of one of the three extracts is not set in the required range of the invention, the sterilization effect of the whole disinfectant is influenced, and the comparison experiment results of the comparative example 1 and the embodiment of the invention also show that the sterilization synergistic effect of the extracts is more remarkable.
In contrast, in comparative example 6, phosphoric acid was not used, which resulted in a great decrease in permeability to bacteria at the time of sterilization, resulting in a lower killing effect than in the examples.
Experimental example 2 toxicology detection experiment
Example 1, example 5, example 9 and comparative example 3 were selected as sample materials.
1. Acute oral toxicity test
SPF grade KM mice supplied by the Biotechnology Co., ltd. Of Pearl sea are selected for acute oral toxicity test.
6.2502 parts of sample raw material is weighed, pure water is added to prepare 25mL of solution, then 5000m parts/kg of body weight dose is fed to a fasted mice at one time, the mice are continuously observed for 14 days, weighing is carried out every 7 days, and general anatomical observation is carried out on animals which survive after death and observation are finished, and the test results are shown in the following table 3:
table 3 body weight observation record
The infected animals do not see any toxic symptoms and toxic death within 14 days, the animal body weight is not abnormal, the surviving animals are generally dissected after the test is finished, no abnormality is found, and the animal is actually nontoxic according to the evaluation rules of the disinfection technical Specification (2002 edition).
In the experimental results of the above examples and comparative examples, the mice of comparative example 3 grew significantly more slowly than the other groups, and were found to have slight toxicity by assaying the mice in vivo, but the mice contained therein had reduced physical condition within the following days although they did not die, and had a certain effect on the body although they recovered to normal after detoxification. Therefore, in order to ensure low toxicity of the disinfectant, the added kudzuvine root and lucid ganoderma can be matched with other substances to remove toxicity so as to achieve good sterilization effect of the disinfectant.
2. Acute inhalation toxicity test
SPF grade KM mice supplied by the Biotechnology Co., ltd. Of Pearl sea were selected for acute inhalation toxicity test.
Selecting 1.0mL of sample raw material as a test solution, atomizing and taking the weighed mice for 2 hours, recovering residual liquid, calculating the contamination concentration, observing each mouse at least once within 30 minutes after contamination, continuously observing for 14 days, weighing every 7 days, and performing general anatomical observation on the animals which die and survive after the observation is finished, if abnormality of tissues and organs is found, further performing histopathological examination, wherein the test results are shown in the following table 4:
table 4 body weight observation record
As is evident from the test, the mice grew normally during the period of exposure to the toxin and during the period of observation, and no death or abnormal symptoms were observed, and after the test, all mice were generally dissected and no abnormality was observed by naked eyes, and the toxicity was classified as practically nontoxic according to the evaluation rules of the disinfection technical Specification (2002 edition), except that the comparative example 3 had a slow weight gain compared with other examples, and may also have a relation with the weight affected by some toxicity remaining in the body.
3. Multiple complete skin irritation test
The skin irritation test was performed with normal grade FMMU guinea pigs supplied by the eastern Xinhua laboratory animal farm in the city of guangzhou.
Example 1, example 5, example 9 and comparative example 3 were selected as sample materials, which were used without treatment, and physiological saline was selected as a control.
Shaving the two sides of the back of a guinea pig, checking skin conditions, coating a sample raw material on one side for testing, coating normal saline on the other side for comparison, cleaning with warm water after coating for 4 hours, removing residues, coating once a day, continuously coating for 14 days, and simultaneously testing two guinea pigs corresponding to each sample raw material, wherein the test results are shown in the following table 5:
table 5 skin irritation response score
As a result of the above-described test, the average score of each guinea pig per day was 0, and the skin irritation intensity was classified according to the "disinfection technical Specification" (2002 edition) and it was judged to be of no irritation.
4. Acute eye irritation test
And selecting common New Zealand rabbits provided by the Guangzhou white cloud area Dragon Guixing family animal farm for acute eye irritation test.
Two eyes of New Zealand rabbits are checked, the test is carried out after one day of the check, the sample raw material is sucked, the new Zealand rabbits are dripped into conjunctival sac of one side of the New Zealand rabbits, after 4s are closed, 30s, physiological saline is used for flushing, the other eye is dripped into physiological saline to be used as a control, the observation is carried out, the injury and recovery condition of conjunctiva, iris and cornea of the New Zealand rabbits are observed by naked eyes 1h, 24h and 72h,7 days, 14 days or 21 days after the dripping, if no stimulus reaction occurs in 72h, or the eye stimulus reaction is completely recovered on 7 th or 14 th day, the test can be terminated in advance, and the test results are shown in Table 6:
TABLE 6 eye irritation response score
According to the test results, only example 9 showed some degree of conjunctival edema and conjunctival congestion when the test was performed, but it was soon recovered, and the eye irritation was not recognized by comparing the average score of cornea damage, iris damage, conjunctival congestion and conjunctival edema of New Zealand rabbits in the acute eye irritation test with the eye irritation response classification standard of "disinfection technical Specification" (2002 edition).
In comparative example 3, however, since the radix Puerariae powder and the Ganoderma lucidum powder were not added, a small amount of toxicity was possibly remained, and thus a certain stimulus was generated to the iris and conjunctiva, resulting in that the New Zealand rabbits required a long time to recover.
5. Mouse bone marrow multipotent erythrocyte micronucleus test
SPF grade KM mice supplied by the Biotechnology Co., ltd. Of Pearl sea Bai Shitong were selected for the mouse bone marrow multi-dye erythrocyte micronucleus test.
6.2530 parts of sample is taken, and pure water is added to prepare 25.0mL of solution;
the solution was formulated as acute oral toxic LD in mice 50 The contamination is carried out by > 5000m parts per kg body weight, then pure water (contamination according to 5000m parts per kg body weight) is selected as a negative control, and a pure water solution of cyclophosphamide (contamination according to 40m parts per kg body weight, contamination by intraperitoneal injection) is selected as a positive control;
the number of mice selected from the male and female groups is 25, and the mice are randomly divided into 5 groups, namely a positive control group, a negative control group, an example 1 group, an example 5 group and an example 9 group, and ten mice in each group are male and female participants.
Oral gavage for 30h contamination method is adopted for each group of mice;
killing a mouse after dislocation of cervical vertebra, removing femur, removing tissues such as muscle, cutting femur, sucking 0.1mL of serum from an exposed marrow cavity by using a syringe, flushing the marrow cavity, performing conventional smear, naturally airing, fixing in methanol solution, and flushing and airing by using PBS solution with pH of=6.8 after being dyed by Gitiemsa;
1000 multi-stained erythrocytes (PCEs) were observed per animal, the number of PCEs with micronuclei was recorded, expressed in thousandths, and in addition, 200 PCEs were counted simultaneously with the NCE seen, the PCE/NCE ratio was calculated, and the experimental results are shown in Table 7:
TABLE 7 results of mice bone marrow multiple-stained red blood cell micronucleus test
Note that: (1) micronucleus cell rate (%) was counted in units of groups and expressed as mean ± standard deviation.
(2) P-values were analyzed for significant differences between other groups and negative control groups.
(3) 200 PCEs are counted, and the NCEs found are simultaneously counted to obtain PCEs/NCEs.
From the above experimental analysis, the micronucleus cell rates of the mice in example 1, example 5 and example 9 were all not significantly different from those of the shadow control group (P > 0.05), and micronucleus was not generated according to the evaluation rules of "sterilizing technical Specification" (2002 edition).
As is clear from the above experiments, the disinfectant prepared in the examples of the present invention meets the national specification of disinfection technical Specification (2002 edition), and compared with the examples, the comparative example 3 does not use radix Puerariae powder and Ganoderma powder, which may have a certain toxicity during use, resulting in weaker effects after use than the solutions of the examples of the present invention.
Experimental example 3 New coronavirus inactivation experiment
The disinfectants prepared in examples 1 to 10 and comparative examples 1 to 6 were selected and subjected to a killing experiment of a new coronavirus: after the action time is 1min at the experimental temperature of 20 ℃, the detection effect is shown in the following table 8:
TABLE 8 effect of deactivation experiments
As is clear from the above data, the disinfectant prepared within the scope of the present invention has a strong killing ability against a new coronavirus, whereas the disinfectant prepared outside the scope of the present invention has a relatively weak killing ability against a new coronavirus, and the positive control group has a virus titer value not exceeding the national regulation, which is not marketable, whereas the disinfectant prepared from the commercial raw materials, although it can also have a killing effect at the end, is likely to be infected within 0.5min, has a short time, but still has a certain risk, and the killing effect is significantly lower than that of the present invention.
Meanwhile, after comparative analysis of the raw materials of comparative example 2 and example 1 by thin layer chromatography, it was found that the purity of the prepared acorn extract of example 1 was 99% which contained a part of the extract impurities, whereas the purity of the acorn extract of comparative example 2 was 90%, which was significantly lower than that of the acorn extract of the present invention, and thus the effect of the prepared disinfectant was better in the case that the raw materials prepared by the present invention were excellent.
Experimental example 4 identification of neutralizing agent and continuous antibacterial test
1. Neutralization agent identification test
Example 1, example 6 and example 7 were selected as sample materials and identified using D/E broth as neutralizing agent, and the test results are shown in table 9 below:
table 9 neutralizer identification data
From the above test data, the test results were found to meet all requirements specified by the evaluation, and the measured neutralizer was found to be acceptable.
2. Continuous antimicrobial test
Samples of the control group (physiological saline) and the test group, which selected example 1, example 6, example 7 and comparative example 4, comparative example 5 as sample materials, were tested after being stored at room temperature for a duration (6 h) specified in the long-acting antibacterial instruction, and the test results are shown in the following table 10:
table 10 antibacterial test data
From the above analysis results, it is found that the disinfectant of the present invention has a good continuous sterilization effect within a predetermined period of time (6 h), and it is found from the results that the effect of example 7 of the present invention is lower than that of example 1 in sterilization because the single-chain quaternary ammonium salt used in example 1 of the present invention is dodecyldimethylbenzyl ammonium chloride and the single-chain quaternary ammonium salt used in example 1 of the present invention is a mixture of dodecyldimethylbenzyl ammonium chloride and dodecyldimethylbenzyloxy ethyl ammonium bromide.
In contrast, in comparative example 4, since trehalose was not added, the prepared disinfectant had no good stability, was volatilized or denatured during storage, and thus the disinfection effect was drastically reduced, whereas in comparative example 5, since maltose was added too much, other raw materials were not allowed to function due to the stability of the maltose, resulting in a low disinfection effect, which also could indicate that each raw material and component was indispensable for the preparation in the present invention.
Experimental example 5 sterilizing Effect
Test groups the disinfectants prepared in example 1, example 2, example 8 and comparative example 6 were used as sample materials, and the control group was tested using physiological saline as sample materials.
1. Disinfection field test on skin
The sample materials were directly applied to the skin and the test was repeated three times with the results shown in table 11 below:
surface 11 disinfectant has effect of killing natural skin bacteria
According to the test data, after the disinfectant is acted for 1min, the average sterilization logarithmic value of the disinfectant for natural bacteria in a skin disinfection field test is more than 1.59, and the disinfectant meets the standard requirement that the sterilization logarithmic value is more than or equal to 1.00. It can also be seen from the data that the effect of example 8 in the present invention is weaker than that of example 1, which is that the photocatalyst used in example 8 is not doped with trace elements, which results in the effect thereof being weaker than that of example 1 of the present invention.
In contrast, comparative example 6 was not added with phosphoric acid, and thus the effect was low during sterilization, and the sterilization effect of the present invention was not achieved.
2. Sterilization field test on other surfaces
The wood tabletop was selected as a sterilization object, and the sample materials were directly applied to the wood tabletop, and the test was repeated three times, and the results are shown in table 12 below:
disinfecting effect of table 12 disinfectant on wooden table top
According to the test, after the wood tabletop is disinfected for 1min, the average killing logarithmic value of natural bacteria is more than 1.81, and the standard requirement that the killing logarithmic value is more than or equal to 1.00 is met.
In contrast, in comparative example 6, the phosphoric acid was not added, so that the lipid of bacteria could not be broken through well during use, and the bacteria could be killed more deeply, resulting in poor killing effect, which could indicate that the raw materials of the present invention are mutually matched.
Through the above experimental examples, it is clear that the raw materials and parts used in the invention are extremely important, and meanwhile, the preparation method of the raw materials is also particularly important, and only the raw materials prepared by the preparation method of the invention are used for preparing the disinfectant, so that a good disinfection effect can be achieved.
Finally, it is to be understood that the above embodiments are merely exemplary embodiments employed for the purpose of illustrating the principles of the present invention, however, the present invention is not limited thereto. Various modifications and improvements may be made by those skilled in the art without departing from the principles and spirit of the invention, and such modifications and improvements are also considered within the scope of the invention.
Claims (9)
1. An environment-friendly disinfectant capable of rapidly inactivating bacteria and viruses is characterized by being prepared from the following raw materials: 60-80 parts of single-chain quaternary ammonium salt, 10-15 parts of acorn extract, 10-15 parts of polygonum orientale extract, 2-8 parts of Galium aparine extract, 2-8 parts of photocatalyst, 5-10 parts of phosphoric acid, 2-8 parts of radix puerariae powder, 1-5 parts of ganoderma lucidum powder, 1-5 parts of trehalose and 1-5 parts of maltose;
the preparation method of the disinfectant comprises the following steps:
mixing single-chain quaternary ammonium salt with phosphoric acid, photocatalyst, acorn extract, polygonum tinctorium extract and Galium aparine extract, stirring, adding Ganoderma powder, radix Puerariae powder, trehalose, and maltose, heating and stirring to obtain target product;
the preparation method of the acorn extract comprises the following steps:
washing acorn, drying, grinding into acorn powder, adding diethyl ether solution for leaching, carrying out water bath reflux to obtain leaching liquor, concentrating the leaching liquor, adding the leaching liquor for extraction and degreasing, then adding the leaching liquor into boiling water solution for concentrating to paste, then adding acetone for dissolving, and continuously extracting and filtering to obtain the acorn extract;
the preparation method of the cortex polygoni orientalis extract comprises the following steps:
adding methanol solution into cortex polygoni chinensis, performing osmotic extraction to obtain an extracting solution, concentrating the extracting solution to paste, adding water, selecting dichloromethane for extraction, recovering under reduced pressure, and eluting by adopting silica gel column chromatography to obtain the cortex polygoni chinensis extract;
the photocatalyst is a nano silicon dioxide photocatalyst; the nano silicon dioxide photocatalyst contains rare earth elements and boron elements, wherein the rare earth elements are at least one of lanthanum, cerium, praseodymium or neodymium.
2. A disinfectant according to claim 1, which is prepared essentially from the following raw materials: 65-78 parts of single-chain quaternary ammonium salt, 12-14 parts of acorn extract, 12-14 parts of polygonum perfoliatum extract, 3-7 parts of Galium aparine extract, 3-7 parts of photocatalyst, 6-9 parts of phosphoric acid, 4-7 parts of radix puerariae powder, 2-4 parts of ganoderma lucidum powder, 2-4 parts of trehalose and 2-4 parts of maltose.
3. A disinfectant according to claim 1, which is prepared essentially from the following raw materials: 70 parts of single-chain quaternary ammonium salt, 13 parts of acorn extract, 13 parts of polygonum multiflorum extract, 5 parts of Galium verum extract, 5 parts of photocatalyst, 7 parts of phosphoric acid, 5 parts of kudzuvine root powder, 3 parts of lucid ganoderma powder, 3 parts of trehalose and 3 parts of maltose.
4. The disinfectant according to claim 1, wherein the single-chain quaternary ammonium salt is one or a combination of two of dodecyldimethylbenzyl ammonium chloride and dodecyldimethylbenzyloxyethyl ammonium bromide.
5. A disinfectant according to claim 1, wherein the phosphoric acid is esterified prior to mixing.
6. The disinfectant according to claim 1, wherein the drying temperature is 85-95 ℃ and the extract is selected from one or a combination of carbon tetrachloride and ethylene oxide.
7. The disinfectant according to claim 1, wherein the silica gel column is 250-300 mesh in size by chromatography eluting while the extract of Polygonum tinctorium is prepared by depressurizing to 40-48 ℃ and 1-1.5 MPa.
8. The disinfectant according to claim 1, wherein the photocatalyst preparation method comprises the steps of:
adding rare earth elements and boron elements into the nano silicon dioxide photocatalyst, adding water, stirring, roasting in a muffle furnace, and crushing to obtain the modified nano silicon dioxide.
9. The disinfectant according to claim 8, wherein the photocatalyst is prepared at a stirring speed of 1100-1200rpm and the muffle furnace is at a temperature of 550-650 ℃.
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