CN111418602A - No-clean disinfectant and preparation method and application thereof - Google Patents
No-clean disinfectant and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a no-clean disinfectant, which consists of polyhexamethylene guanidine hydrochloride, glycyrrhizic acid, glycerol and purified water. The washing-free disinfectant disclosed by the invention utilizes polyhexamethylene guanidine hydrochloride and a virus protein receptor ACE2) inhibitor glycyrrhizic acid as main sterilization and antiviral components, and plays a role in sterilization and antivirus. The product overcomes the defects of strong irritation, slight toxicity and the like of most disinfectants sold in the market, becomes a high-efficiency environment-friendly disinfectant with broad-spectrum sterilization and disinfection, high action speed, good stability, low toxicity, no corrosiveness and small irritation, particularly easily forms a layer of 'liquid gloves' on hands of a human body when the product is used, plays a role in disinfection and protection, and has wide application prospect.
Description
Technical Field
The invention relates to a disinfectant, a preparation method and application thereof, in particular to a washing-free virus-killing disinfectant, and a preparation method and application thereof. Belongs to the technical field of chemical disinfectants.
Background
Hospitals, banks, supermarkets, organs, enterprises and public institutions, movie theaters, troops, entertainment venues, schools, kindergartens, families, hotels, restaurants, airports, docks, railway stations and other places are very easy to cause the spread of bacteria and viruses, thereby causing public cross infection. The presence of harmful microorganisms can cause a number of diseases and, in severe cases, can endanger human life. Disinfection and sterilization are effective measures for maintaining the life safety of human beings. In daily life, various disinfectants are often used.
At present, the active ingredients of the commonly used disinfectants mainly comprise the following substances: oxidizing substances, aldehydes, phenols, alcohols, alkali salts, halogens, surfactants, and the like, but all have disadvantages. The ideal disinfectant has the characteristics of wide bactericidal spectrum, strong bactericidal capacity, high action speed, good stability, low toxicity, corrosiveness, small irritation (no toxicity, no residue, no corrosion or no irritation), easy water dissolution, safety to human and animals, low price, easy obtainment, low environmental pollution degree and the like. Therefore, there is an urgent need to develop such disinfectants that are more rapidly effective and non-toxic.
Railings, door handles, handrails in a carriage, money bills and the like which are used for daily hand holding or hand touching are important factors causing public cross infection. In this case, the leave-on disinfectant takes place at the end of life. The hand-washing-free disinfectant can save precious water resources while ensuring health of people, and does not need to use towels, water and soaps. At present, related washing-free disinfectants have been reported or products, but the principle of sterilization and disinfection of polyhexamethylene guanidine hydrochloride is utilized, and glycyrrhizic acid and glycerol reagents are matched, so that the defect that the conventional polyhexamethylene guanidine hydrochloride has certain corrosivity to aluminum and carbon steel is overcome, and the prepared high-efficiency environment-friendly disinfectant has the advantages of broad-spectrum sterilization, high acting speed, good stability, low toxicity, no corrosivity and small irritation, is particularly easy to form a layer of 'liquid gloves' on hands of a human body, and has a disinfection and protection effect.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a washing-free virus-killing disinfectant and a preparation method and application thereof.
The washing-free disinfectant disclosed by the invention consists of polyhexamethylene guanidine hydrochloride, glycyrrhizic acid, glycerol and purified water, and is characterized in that: the pH value of the no-clean disinfectant is 5.5-7, the components of the no-clean disinfectant are calculated by 100ml of solution, and the no-clean disinfectant comprises 1.5 ml-3 ml of polyhexamethylene guanidine hydrochloride solution with the mass concentration of 25%, 0.05 ml-0.2 ml of glycyrrhizic acid, 3 ml-6 ml of glycerol and the balance of purified water.
Wherein: the pH value of the no-clean disinfectant is preferably 6.0-6.8, and the components of the no-clean disinfectant are preferably 2ml of a 25% polyhexamethylene guanidine hydrochloride solution, 0.1ml of glycyrrhizic acid, 5ml of glycerol and 92.9ml of purified water in terms of 100ml of solution.
The preparation method of the no-clean disinfectant comprises the following steps:
(1) adding 70-80% of purified water and 25% of polyhexamethylene guanidine hydrochloride solution at the mass concentration at the formula amount into a stirring tank at normal temperature, and stirring until the mixture is completely uniform for later use;
(2) adding glycyrrhizic acid and glycerol in formula amount and purified water in balance at normal temperature, and stirring uniformly;
(3) testing pH value, determining whether the pH value reaches 5.5-7, and filling if the pH value is between 5.5-7; if the pH value does not reach the standard, the pH value is adjusted by citric acid.
The preparation method of the no-clean disinfectant comprises the following steps:
(1) adding 70ml of purified water and 2ml of poly (hexamethylene guanidine) hydrochloride solution with the mass concentration of 25% into a stirring tank at normal temperature based on 100ml of prepared disinfectant, and stirring until the mixture is completely uniform for later use;
(2) adding glycyrrhizic acid 0.1ml, glycerin 5ml and purified water 22.9ml at normal temperature, and stirring;
(3) testing the pH value, determining whether the pH value reaches 6.0-6.8, and filling if the pH value is 6.0-6.8; if the pH value does not reach the standard, the pH value is adjusted by citric acid.
The washing-free disinfectant disclosed by the invention is applied to antibiosis and antivirus.
Wherein: the bacteria are preferably Escherichia coli 8099, Staphylococcus aureus ATCC6538, Pseudomonas aeruginosa ATCC15442 and Candida albicans ATCC 10231; the virus preferably refers to a novel coronavirus, specifically a novel coronavirus SARS-CoV-2 and a novel coronavirus 2019-nCoV.
The no-clean disinfectant provided by the invention can be applied to sterilization and disinfection of skin, hand disinfection, air disinfection and other various environmental aspects.
In the components of the no-clean disinfectant disclosed by the invention, polyhexamethylene guanidine hydrochloride is a sterilization component, methyl guanidine has high activity per se, so that a polymer is electropositive, the sterilization mechanism of the no-clean disinfectant is the combined action of chemistry and physics, firstly, the no-clean disinfectant can strongly act with a microbial cell membrane which usually has negative charges, and the ion gradient necessary for storing energy and transporting nutrients of the microbe is destroyed, so that the cell membrane is broken; meanwhile, the substance is positive and can be easily adsorbed on the surfaces of various negatively charged bacteria and viruses, the division function of the bacteria and the viruses is inhibited, the bacteria and the viruses lose the reproductive capacity, and the polymers form to block the breathing channel of the microorganisms, so that the microorganisms are suffocated. Therefore, at an extremely low concentration (10ppm), the polyhexamethylene guanidine hydrochloride can kill cotton fabric bacteria catalase and fungi, acanthamoeba keratitis pathogenic bacteria, escherichia coli, staphylococcus aureus and candida albicans, skin natural bacteria, inactivated viruses, bacterial spores, bacterial propagules and the like. Glycyrrhizic acid is a novel inhibitor of a coronavirus (SARS-CoV-2) protein receptor ACE2, can be combined with angiotensin converting enzyme 2(ACE2) to block combination of a virus S protein and an ACE2 receptor, and reduces invasion of viruses into epidermal cells of a human body, so that harm of the viruses to the human body is reduced. Neurosurgery of the medical college of Stanford university in USA and the college of traditional Chinese medicine of hong Kong university in United states together published papers, and it was also confirmed that glycyrrhizic acid can bind to ACE2, and thus it can be concluded that glycyrrhizic acid is a potential anti-SARS-CoV-2 compound, and has important value for preventing novel coronavirus (SARS-CoV-2) infection and treating SARS. In medicine, glycerin is generally used for preparing various preparations, solvents, moisture absorbents, antifreezes and sweeteners, and formulating ointments or suppositories for external use, etc. Because glycerin has water absorption, it is commonly used as a moisture absorbent and moisturizer for cosmetics, foods, and textiles. The glycyrrhizic acid and the glycerol are added into the no-clean disinfectant, and the polyhexamethylene guanidine hydrochloride is coordinated, so that the product is convenient to use and easy to paint, can form a better 'film' after being dried, forms 'invisible gloves' on the surfaces of objects or hands, and plays a role in inhibiting bacteria for a long time.
The invention provides a washing-free virus-killing disinfectant and a preparation method and application thereof. Wherein the disinfectant is a hydrophilic solution consisting of polyhexamethylene guanidine hydrochloride, glycyrrhizic acid and glycerol. The disinfectant has the remarkable advantages that: 1. broad-spectrum antibiosis; 2. the action speed is high, and the effect is achieved within 5 minutes; 3. the effect lasts for 4-5 hours, and the maximum time can reach 6 hours; 4. the liquid gloves are formed, have the functions of gentle skin care, preventing skin damage and moistening and caring hand skin; 5. completely non-toxic, degradable and good in environmental protection property; 6. is suitable for disinfection and sterilization of object surfaces, water, air and other environments.
Detailed Description
The present invention will be described in detail with reference to specific examples. The following examples are merely illustrative of the preferred embodiments of the present invention and are not intended to limit the present invention in any way, and any simple modifications, equivalent changes and modifications made to the embodiments according to the technical spirit of the present invention fall within the scope of the technical solution of the present invention.
Example 1: preparation of washing-free virus-killing disinfectant
Taking the preparation of 100ml of washing-free virus-killing disinfectant as an example:
(1) adding 70ml of purified water and 2ml of poly (hexamethylene guanidine hydrochloride) solution with the mass concentration of 25% into a stirring tank at normal temperature, and stirring until the solution is completely uniform for later use;
(2) adding glycyrrhizic acid 0.1ml, glycerin 5ml and purified water 22.9ml at normal temperature, and stirring;
(3) testing the pH value, determining whether the pH value reaches 6.0-6.8, and if the pH value is 6.0-6.8, preparing 100ml of the washing-free virus-killing disinfectant; if the pH value does not reach the standard, the pH value is adjusted by citric acid.
Example 2: evaluation of Disinfection by disinfectant
2.1 killing of microorganisms test
2.1.1 preparation of bacterial suspensions and pellets
The strains are Escherichia coli 8099, Staphylococcus aureus ATCC6538, Pseudomonas aeruginosa ATCC15442 and Candida albicans ATCC 10231.
2.1.2 tests were carried out according to the sterilization test method specified in the Disinfection technical Specification 2002 edition, the results of which are shown in Table 1.
Table 1: evaluation of Effect of killing microorganisms
2.2 disinfectant on-site experiment on hand disinfection
2.2.1 at the site of use, subjects were randomly selected. The test is not less than 30 people.
2.2.2 before disinfection, after the two hands of the testee are rubbed fully, the left fingers of the testee are closed, the testee is soaked in a test tube containing 10ml of diluent by using a sterile cotton swab, after the tube wall is squeezed to be dry, the fingers of the five fingers are bent to the roots of the fingers, and the two fingers are rubbed repeatedly for 2 times, and the cotton swab is rotated once every rubbing. After sampling, the sampling end of the swab was aseptically cut into a tube of neutralizing reagent to serve as a positive control sample.
2.2.3 the right hand is disinfected according to the using method of the disinfectant, and the acting time of the sanitary disinfection of the hand is generally set to be 1 min. After disinfection, the neutralizing agent is used to replace the diluent, and the natural bacteria remained on the right hand of the subject is sampled once in the same way as the positive control group and is used as a test group sample.
2.2.4 the unused neutralizer of the same batch, 1.0ml of each diluent and 1-2 parts of cotton swab are respectively used as negative control group samples.
2.2.5 taking 1.0ml of each sample of the test group, the positive control group and the negative control group, inoculating the samples into the plates by an agar pouring method, inoculating 2 plates into each sample, putting the plates into a 37 ℃ incubator for culturing for 48 hours, and observing the final result.
3.3 evaluation assay
The positive control group should have more bacteria growing, the negative control group should have sterile growth, and the disinfection is qualified when the average killing log value of the natural bacteria on hands of 30 persons is more than or equal to 1.00. The bactericidal effect is shown in table 2.
Table 2: sterilizing effect after 30 people spray disinfectant
Example 3: disinfectant protection evaluation
3.1 antibacterial circle method for testing bactericidal and antibacterial effects of disinfectant
3.1.1 preparation of 100m L L B liquid medium 2 bottles (yeast extract 5 g/L, tryptone 10 g/L10 g/L, agar powder 15 g/L), autoclaving at 121 ℃ for 20min, cooling, inoculating 1 strain on the inclined plane of Escherichia coli and 1 strain on the inclined plane of Staphylococcus aureus, shaking up, and culturing at 37 ℃ for 1-2 days until the culture is turbid.
3.1.2 in the clean bench, put into 100m L about 45 ℃ nutrient agar medium, shake, then pour the culture medium into the dish, wait to solidify, pour 4 dishes, suck 0.2m L of colibacillus (8099) suspension and 0.2ml of staphylococcus aureus (ATCC6538) with the aseptic pipette, spread in the culture medium.
3.1.3 taking sterile circular filter paper sheets (5mm) completely soaked by the disinfectant, respectively and horizontally placing the sterile circular filter paper sheets into a culture dish containing escherichia coli and a culture dish containing staphylococcus aureus, carrying out static culture at 37 ℃, and taking out the sterile circular filter paper sheets for 24h/48h/72h to determine the size of a bacteriostatic zone. The results are shown in Table 3.
Table 3: zone of inhibition test
According to the size of the inhibition zone, the disinfectant provided by the invention can effectively inhibit the growth of escherichia coli and staphylococcus aureus, and can continuously and stably play a role in inhibiting bacteria.
Example 4: batch preparation of washing-free virus-killing disinfectant
1) Adding purified water and a formula amount of 25% polyhexamethylene guanidine hydrochloride solution at the mass concentration of 2% into a stirring tank at normal temperature, and stirring until the mixture is completely uniform for later use;
2) adding purified water and glycyrrhizic acid with formula amount to final concentration of 0.1% and glycerol to final concentration of 5% at room temperature, and stirring;
3) testing pH value, if the pH value is not between 5.5-7, adjusting pH value to 5.5-7 with citric acid, and bottling.
Example 5: application of washing-free virus-killing disinfectant in novel coronavirus nucleic acid detection verification
In order to detect the disinfection effect of the no-clean virus-killing disinfectant on viruses, 5 nucleic acid samples extracted from novel SARS-CoV-2 nucleic acid positive patients in infectious disease hospitals in Jinan City are utilized, and 1 volume percent of the no-clean virus-killing disinfectant is added into each SARS-CoV-2 positive RNA and mixed uniformly. And a novel coronavirus (SARS-CoV-2) nucleic acid detection kit (constant temperature nucleic acid amplification method) (Shandongbaobo Biotechnology Co., Ltd., batch No. 202002001) is used together with a nucleic acid rapid enhancement solution (Shandongbaobo Biotechnology Co., Ltd., batch No. 202002094) to detect the RNA content in each case of SARS-CoV-2 positive RNA, and to verify whether the low-concentration virus RNA exists after disinfection. The specific operation method comprises the following steps:
5.1 Add 2.5. mu. L nucleic acid template to the reaction tube (5. mu. L for positive and negative controls).
5.2 Add 2.5 μ L B Buffer to each reaction tube, cover the tube cap (for many reactions, it is recommended to add B Buffer to the inner side of the cap of the reaction tube), reverse the reaction tube from top to bottom for 8-10 times, mix well, throw (or fast centrifuge) the reaction solution to the bottom of the tube after mixing well, transfer to the amplification area.
5.3 Using Roche L ightCycler480, the reaction solution was transferred to a Roche-adapted dedicated reaction tube and subjected to amplification reaction.
5.4 nucleic acid amplification
| Step (ii) of | Temperature of | Time of day | Number of cycles |
| ① incubation | 42℃ | 1min | 1cycle |
| ② amplification/detection of fluorescence | 42℃ | 30s | 40ycle |
5.5, uniformly mixing the mixed solution by using a pipettor in a blowing and sucking or vortex mode, and instantaneously centrifuging to remove bubbles; can be premixed and then packaged; the operation on ice is not needed. And (3) carrying out reaction on the uniformly mixed system on a qPCR instrument.
Reaction procedure: FAM fluorescence was detected every 30s at 48 ℃ for 20 min. For the reinspection of negative (Ct is more than or equal to 40) samples, the reaction time is recommended to be set to 60min, and the sensitization effect is enlarged.
5.6 more than 20 groups of fluorescence values are collected from each sample, and straight line fitting is carried out by taking the cycle times as an X axis and the fluorescence values as a Y axis to obtain a straight line slope K value. (negative for K value < 500)
Table 4: positive RNA sample nucleic acid detection result
| Sample numbering | Ct before adding no-washing disinfectant | Ct after adding washing-free disinfectant | Adding washing-free disinfectant K |
| 1 | 33 | Not detected out | 50 |
| 2 | 30 | Not detected out | 55 |
| 3 | 28 | Not detected out | 35 |
| 4 | 34 | Not detected out | 46 |
| 5 | 31 | Not detected out | 73 |
The results show that the results of PCR detection of the nucleic acid positive sample (SARS-CoV-2) after the treatment of the washing-free disinfection solution and the detection after the subsequent addition of the enhancement solution to improve the sensitivity are negative. It shows that the washing-free disinfectant of the present invention has strong effect of destroying SARS-CoV-2 and may be used in killing SARS-CoV-2. This provides a good basis for the virus protection work.
Claims (7)
1. A washing-free disinfectant consists of polyhexamethylene guanidine hydrochloride, glycyrrhizic acid, glycerol and purified water, and is characterized in that: the pH value of the no-clean disinfectant is 5.5-7, the components of the no-clean disinfectant are calculated by 100ml of solution, and the no-clean disinfectant comprises 1.5 ml-3 ml of polyhexamethylene guanidine hydrochloride solution with the mass concentration of 25%, 0.05 ml-0.2 ml of glycyrrhizic acid, 3 ml-6 ml of glycerol and the balance of purified water.
2. The no-clean disinfectant as set forth in claim 1, wherein: the pH value of the no-clean disinfectant is 6.0-6.8, and the components of the no-clean disinfectant are calculated by 100ml of solution, and the no-clean disinfectant comprises 2ml of poly (hexamethylene guanidine hydrochloride) solution with the mass concentration of 25%, 0.1ml of glycyrrhizic acid, 5ml of glycerol and 92.9ml of purified water.
3. The method for preparing the no-clean disinfectant as claimed in claim 1 or 2, comprising the steps of:
(1) adding 70-80% of purified water and 25% of polyhexamethylene guanidine hydrochloride solution at the mass concentration at the formula amount into a stirring tank at normal temperature, and stirring until the mixture is completely uniform for later use;
(2) adding glycyrrhizic acid and glycerol in formula amount and purified water in balance at normal temperature, and stirring uniformly;
(3) testing pH value, determining whether the pH value reaches 5.5-7, and filling if the pH value is between 5.5-7; if the pH value does not reach the standard, the pH value is adjusted by citric acid.
4. The method for preparing a no-clean disinfectant as claimed in claim 3, wherein the method comprises the following steps:
(1) adding 70ml of purified water and 2ml of poly (hexamethylene guanidine) hydrochloride solution with the mass concentration of 25% into a stirring tank at normal temperature based on 100ml of prepared disinfectant, and stirring until the mixture is completely uniform for later use;
(2) adding glycyrrhizic acid 0.1ml, glycerin 5ml and purified water 22.9ml at normal temperature, and stirring;
(3) testing the pH value, determining whether the pH value reaches 6.0-6.8, and filling if the pH value is 6.0-6.8; if the pH value does not reach the standard, the pH value is adjusted by citric acid.
5. The use of the leave-on disinfectant of claim 1 or 2 for antimicrobial and antiviral purposes.
6. Use according to claim 5, characterized in that: the bacteria are Escherichia coli 8099, Staphylococcus aureus ATCC6538, Pseudomonas aeruginosa ATCC15442 and Candida albicans ATCC 10231; the virus is a novel coronavirus.
7. Use according to claim 6, characterized in that: the virus refers to a novel coronavirus SARS-CoV-2 and a novel coronavirus 2019-nCoV.
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| CN113647417A (en) * | 2021-08-18 | 2021-11-16 | 江苏耀研达科技发展有限公司 | Ecological environment-friendly plant-derived virus and bacterium inactivator, preparation method, inactivation identification method and application thereof |
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Application publication date: 20200717 |