CN111742925B - Environment-friendly long-acting washing-free sterilizing disinfectant and preparation and use methods thereof - Google Patents
Environment-friendly long-acting washing-free sterilizing disinfectant and preparation and use methods thereof Download PDFInfo
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Abstract
The invention provides an environment-friendly long-acting no-clean sterilizing disinfectant and a preparation method and a use method thereof. The sterilizing disinfectant comprises the following raw material components in percentage by mass: ACS: 1-50%; and (3) chitosan: 30-95%; low odor fast volatilization accelerator: 0.1-5%; water: and (4) the balance. Wherein: the ACS is added in three ACS aqueous solutions with different mass percentage concentrations, and specifically comprises ACS aqueous solutions with the concentrations from high to low, namely a first concentration, a second concentration and a third concentration; the chitosan consists of chitosan with a medium deacetylation degree of 80-85% and chitosan with a high deacetylation degree of 90-95%. The invention provides a killerThe bacteria disinfectant has transparent appearance, no color and no smell, and can kill various bacteria and coronavirus for a long time by 100 times of water diluent; the product does not contain ethanol, active chlorine, free chlorine and the like, and the LD of the product is detected50The value is more than or equal to 5000mg/kg, and the product is safe, environment-friendly and non-toxic; when in use, the product can be directly sprayed into indoor, clothes, skin, livestock and poultry houses, etc.
Description
Technical Field
The invention relates to the technical field of disinfectant, in particular to an environment-friendly washing-free sterilizing disinfectant.
Background
The use of sterilizing and disinfecting liquid has become one of the necessary protective means in people's life. The current common disinfectant mainly comprises: 75 percent of medical alcohol, 0.05 percent of sodium hypochlorite solution (84 main components of disinfectant), hydrogen peroxide/peroxyacetic acid disinfectant and the like, wherein the 9 disinfectant can effectively kill coronavirus. But has potential safety hazard and environmental destruction. 75% of medical alcohol belongs to flammable and explosive dangerous goods, when the content of ethanol in the air reaches 19% and the temperature is higher than 13 ℃, the air can deflagrate when meeting sparks, so that the alcohol-containing disinfectant can only perform wiping disinfection on skin or appliances; 84 the disinfection solution has strong irritation and corrosiveness, free chlorine released by the disinfection solution has toxicity, and organisms can inhale or ingest the disinfection solution harmfully; peroxide disinfectant has strong irritation to skin and reacts violently with alkali, so that safety risk exists.
Acid Calcium Sulfate (ACS) is a novel bactericidal and anticorrosive product, is prepared from calcium hydroxide, calcium sulfate and sulfuric acid, and has the characteristics of no color, no odor, easy water solubility, no volatility, stability and low corrosivity. The chemical essence of the method is an acidic solution of a slightly soluble IIA complex, the acid equivalent concentration is equivalent to 98% sulfuric acid (36N), and the strong acidity of ACS can be utilized to break hydrogen bonds of glycoprotein on the surface of an envelope fat membrane outside a virus, destroy the secondary structure and the tertiary structure of protein, and cause the crystalline damage and the biological activity loss of the glycoprotein. The ACS can effectively create a low-corrosivity extremely-low-pH-value environment for inhibiting harmful microorganisms, and further achieve the effects of prolonging sterilization and disinfection. The bactericidal effect of ACS has a large relationship with the acidity, concentration and the like of an acidic solvent, the higher the concentration of the acidic solvent is, the higher the bactericidal rate of ACS is, but the higher the acidity is, the higher the irritation and corrosivity are, and meanwhile, chitosan, which is a product obtained by deacetylating chitin, is a natural antibacterial substance with a wider antibacterial spectrum, and the mechanisms of chitosan sterilization and virus resistance are embodied in three aspects: (1) the chitosan has a polycation structure and is easy to combine with a group with negative charges on the surfaces of pathogenic bacteria cells or viruses, so that the fluidity and the permeability of pathogenic bacteria cell membranes or virus surface envelopes are blocked; (2) interfere with DNA replication and transcription; preventing reverse transcription and translation of RNA (3) blocks normal metabolism of pathogenic bacteria and viruses. But the chitosan alone has long time for sterilization and disinfection and low efficiency.
Aiming at the defects of the single acidic calcium sulfate and chitosan disinfectant, the invention aims to provide the sterilizing disinfectant with the synergistic effect of ACS and chitosan, so as to further reduce the potential low corrosivity of ACS, improve the overall safety, sterilizing effect and environmental protection of the disinfectant and reduce the irritation and corrosivity of the disinfectant. No sterilizing disinfectant adopting the synergistic effect of ACS and chitosan exists in the prior art.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: provides a long-acting environment-friendly washing-free sterilizing disinfectant which has the characteristics of safety, environmental protection, washing-free, high sterilizing rate, long sterilizing sustainable time and the like.
In order to solve the technical problem, the technical scheme adopted by the invention is as follows:
on one hand, the invention provides an environment-friendly long-acting no-clean sterilizing disinfectant, which comprises the following raw material components in percentage by mass:
ACS：1～50％；
and (3) chitosan: 30-95%;
low odor fast volatilization accelerator: 0.1-5%;
water: the balance;
wherein:
the ACS is added in three ACS aqueous solutions with different mass percentage concentrations, and specifically comprises an ACS aqueous solution with a first concentration, an ACS aqueous solution with a second concentration and an ACS aqueous solution with a third concentration; the ACS aqueous solution with the first concentration has the lowest concentration, and the ACS aqueous solution with the third concentration has the highest concentration;
the chitosan consists of chitosan with a medium deacetylation degree of 80-85% and chitosan with a high deacetylation degree of 90-95%.
In the invention, ACS is essentially an acidic solution of a slightly soluble IIA complex (AGIIS) and is prepared from sulfuric acid, calcium sulfate, calcium hydroxide and organic acid, and the pH value of the acidic solution can reach-3 at the lowest; the core value of the corrosion inhibitor is characterized in that the corrosion inhibitor has extremely low corrosivity when the strong acidity is maintained. The group IIA complex AGIIS of ACS has an element arrangement structure similar to that of Ca (OH) 2. The special arrangement structure enables ACS to have strong acidity and simultaneously has extremely low corrosivity and corrosion inhibition.
Preferably, the first and second electrodes are formed of a metal,
the mass percentage concentration of the ACS aqueous solution with the first concentration is 4-6%; the mass percentage concentration of the ACS aqueous solution with the second concentration is 8-10%; and the mass percentage concentration of the ACS aqueous solution with the third concentration is 25-35%.
More preferably still, the first and second liquid crystal compositions are,
the mass percentage concentration of the ACS aqueous solution with the first concentration is 5%; the mass percentage concentration of the ACS aqueous solution with the second concentration is 10 percent; the ACS aqueous solution with the third concentration has a mass percentage concentration of 30%.
Further, the air conditioner is provided with a fan,
and the chitosan with the medium deacetylation degree of 80-85 percent: the mass ratio of 90-95% of chitosan with high deacetylation degree is (1-5): 1.
further, the air conditioner is provided with a fan,
the low-odor quick-volatilization promoter is one or a mixture of two of AMP-95 and AMP-96. Preferably, the low odor fast volatilization promoter is a mixture of AMP-95 and AMP-96.
On the other hand, the invention provides a preparation method of the environment-friendly long-acting no-clean sterilizing disinfectant, which specifically comprises the following steps:
s1, diluting the measured ACS into three ACS aqueous solutions with the concentration from low to high: ACS aqueous solution with first concentration, ACS aqueous solution with second concentration and ACS aqueous solution with third concentration;
s2, mixing and stirring the measured ACS aqueous solution with the first concentration and 90-95% of chitosan with the high deacetylation degree uniformly to obtain a first mixed solution, and then heating to 50 ℃;
s3, adding 80-85% of measured chitosan with medium deacetylation degree into the heated first mixed solution, uniformly stirring, then starting dropwise adding an ACS aqueous solution with a second concentration, completing dropwise adding within 10 minutes to obtain a second mixed solution, and heating to 80 ℃;
s4, adding metered low-odor fast-volatilization accelerator, uniformly stirring, then beginning to dropwise add ACS aqueous solution with a third concentration, completing dropwise addition within 30 minutes, and reacting for 2-8 hours at 70-90 ℃;
and S5, adding the balance of water, stirring, cooling and cooling to obtain the product.
The disinfectant prepared by the invention is transparent in appearance, colorless and tasteless.
Further, in step S1, the measured ACS is divided into three equal volumes, and then diluted into three different ACS aqueous solutions for use.
Further, in the method, the mass percentages of the measured raw materials are as follows:
ACS: 1-50%; and the ACS is added in a manner that the diluted ACS aqueous solution with the mass percentage concentration of 4-6% and the first concentration, the diluted ACS aqueous solution with the mass percentage concentration of 8-10% and the diluted ACS aqueous solution with the third concentration are added in a manner of multiple times; more preferably, the ACS aqueous solution of the first concentration is 5% by mass; the mass percentage concentration of the ACS aqueous solution with the second concentration is 10 percent; the ACS aqueous solution with the third concentration has a mass percentage concentration of 30%.
The sum of the chitosan with medium deacetylation degree and the chitosan with high deacetylation degree is as follows: 30-95%; and the mass ratio of the chitosan with medium deacetylation degree to the chitosan with high deacetylation degree is (1-5): 1.
low odor fast volatilization accelerator: 0.1-5%; preferably, the low odor fast volatilization promoter is AMP-95 (a multifunctional amine adjuvant, chemical name: 2-amino-2-methyl-1-propanol; formula: (CH)3)2C(NH2)CH2OH), AMP-96 (an organic alcohol amine)Class compound) or a mixture of two. More preferably, the low odor fast volatilization promoter is a mixture of two of AMP-95 and AMP-96.
Water: and (4) the balance.
The invention also provides a using method of the environment-friendly long-acting no-clean sterilizing disinfectant or the environment-friendly long-acting no-clean sterilizing disinfectant prepared by the preparation method, and specifically, the environment-friendly long-acting no-clean sterilizing disinfectant is diluted by 100 times by adding water and is used for killing staphylococcus aureus, escherichia coli and/or coronavirus.
The sterilization principle of the disinfectant disclosed by the invention is as follows:
under the acidic condition, due to the existence of a chitosan acetal structure, a beta-1, 4-glycosidic bond of the chitosan can be slowly hydrolyzed to generate chitosan with positive charge and low relative molecular mass, so that the binding force of the chitosan with negatively charged protein and nucleic acid is further enhanced, and the chitosan permeates into a membrane structure to be combined with DNA or RNA in bacteria or viruses, so that the translation, reverse transcription and protein synthesis of the RNA can be inhibited and hindered, and the effects of sterilizing and killing the viruses are achieved. The chitosan, particularly the chitosan with medium deacetylation degree adopted in the invention, is very easy to form a film and plays a role in water retention, and the potential low corrosivity of ACS is further reduced under the protection of the chitosan. Therefore, the sterilization disinfectant with the synergistic effect of the ACS and the chitosan can durably kill and effectively kill the representative staphylococcus aureus and escherichia coli, can effectively inactivate coronavirus, and has the characteristics of safety, harmlessness, no irritation and environmental protection.
According to the invention, the chitosan with a deacetylation degree of 80-85% and the chitosan with a high deacetylation degree of 90-95% are adopted at the same time: 1) as the deacetylation degree of the chitosan is higher, the free amino groups are more, the binding force with lipoid polysaccharide substances is stronger, and the bacteriostasis effect is stronger, the binding sites of the amino groups and the teichoic acid of the chitosan with high deacetylation degree are more, and the binding force is stronger. Taking staphylococcus aureus as an example, the chitosan with high deacetylation degree is more easily combined with rich teichoic acid in the cell wall of staphylococcus aureus, so that the physiological function of the staphylococcus aureus is influenced, the propagation of the staphylococcus aureus is finally inhibited, and the killing effect is realized. 2) Meanwhile, the higher the deacetylation degree of chitosan, the better the water solubility. The chitosan with medium deacetylation degree has better film forming property, larger molecular weight and longer decomposition time, and also has bacteriostatic ability which is slightly weaker than that with high deacetylation degree. Therefore, the main function of the chitosan with medium deacetylation degree in the disinfectant is to ensure the durability of the disinfection effect.
Because chitosan is insoluble in pure water, and the solubility of chitosan is better in weak acid solution, in the preparation method of the invention, the chitosan is firstly dissolved by using ACS aqueous solution with weaker acidity and lower concentration (such as ACS aqueous solution with 5% of mass percentage concentration), and the ACS aqueous solution with higher concentration and third concentration (such as ACS aqueous solution with 30% of mass percentage concentration) has relatively lower pH value and stronger acidity, and is added finally, and the low-odor quick-volatilization accelerant is required to be used for firstly adjusting the pH value and then adding to help mixing and dissolving.
In the preparation method, the control of the reaction time and temperature of each step can make the impression that the transparency of the product and part of chitosan are further hydrolyzed into glucosamine, chitobiose, chitotriose and other substances, and influence the final compatibility and durability effect of the product, and the substances are separated out due to insufficient reaction time; the preparation time is too long, the preparation time process increases the production cost, and the production cost and the product quality can be better ensured at the same time under the reaction, time and temperature of the scheme provided by the invention.
Compared with the prior art, the invention has the following beneficial effects:
1. the killing effect is quick and long-acting. The invention uses inorganic ACS and natural chitosan as raw materials to sterilize, and uses the strong acidity and low corrosivity of ACS and the biocompatibility and antibacterial property of chitosan to prepare the disinfectant product with quick and strong sterilization and disinfection effects, which can effectively kill the representative staphylococcus aureus and escherichia coli and has better inactivation effect on coronavirus.
2. Is safe and environment-friendly. 1) The invention adopts chitosan as the carrier, thus further reducing the corrosivity of ACS; the chitosan in the product can be slowly hydrolyzed and finally dissolved in water, and has no stimulation to skin, and the chitosan which carelessly enters the human body can be gradually decomposed into glucosamine which is normally metabolized, so that the product is safer. 2) Compared with disinfectant containing sodium hypochlorite, the sterilization disinfectant provided by the invention does not contain free chlorine and active chlorine, and is harmless to the environment and human bodies. 3) The sterilization disinfectant does not contain ethanol, and has no flammable and explosive dangers. 4) The sterilizing disinfectant disclosed by the invention is transparent in appearance, colorless and tasteless, free of pungent smell and free of corrosiveness.
3. Is non-toxic and free of washing. LD50 (median lethal dose, LD50 for short) of the sterilizing disinfectant disclosed by the invention indicates that the minimum bacterial count or toxin amount required by killing half of certain animals with certain weight or age through specifying an infection way within a specified time is more than or equal to 5000mg/kg, and the sterilizing disinfectant belongs to a practical non-toxic product. The sterilization disinfectant can be used by wiping, or by dipping or spraying, does not need to be cleaned additionally after being used, and belongs to a no-clean disinfectant.
4. Broad spectrum applicability. The sterilization disinfectant provided by the invention can be directly used for sterilization and disinfection of human bodies, clothes, masks, indoor houses, automobile appliances and the like, and can also be used for sterilization and disinfection of classrooms, elevators, hospitals, hotels, airplane interiors, food production lines, kitchen appliances, restaurants, public facilities, animal houses, poultry houses and the like.
Detailed Description
The invention provides an environment-friendly long-acting no-clean sterilizing disinfectant, and a preparation method and a use method thereof. Wherein:
the environment-friendly long-acting no-clean sterilizing disinfectant provided by the invention comprises the following raw material components in percentage by mass:
ACS：1～50％；
and (3) chitosan: 30-95%;
low odor fast volatilization accelerator: 0.1-5%;
water: and (4) the balance.
Wherein:
the ACS is added in three ACS aqueous solutions with different mass percentage concentrations, and specifically comprises an ACS aqueous solution with a first concentration, an ACS aqueous solution with a second concentration and an ACS aqueous solution with a third concentration; the ACS aqueous solution with the first concentration has the lowest concentration, and the ACS aqueous solution with the third concentration has the highest concentration; in a preferred embodiment, the ACS aqueous solution with the first concentration has a mass percentage concentration of 4-6%; the mass percentage concentration of the ACS aqueous solution with the second concentration is 8-10%; and the mass percentage concentration of the ACS aqueous solution with the third concentration is 25-35%. More preferably, the ACS aqueous solution of the first concentration is 5% by mass; the mass percentage concentration of the ACS aqueous solution with the second concentration is 10 percent; the ACS aqueous solution with the third concentration has a mass percentage concentration of 30%.
The chitosan consists of chitosan with a medium deacetylation degree of 80-85% and chitosan with a high deacetylation degree of 90-95%. As a preferred embodiment, the chitosan with a medium deacetylation degree of 80-85 percent: the mass ratio of 90-95% of chitosan with high deacetylation degree is (1-5): 1.
as a preferred embodiment, the low-odor quick volatilization accelerant is one or a mixture of two of AMP-95 and AMP-96.
The preparation method of the environment-friendly long-acting no-clean sterilizing disinfectant specifically comprises the following steps:
s1, diluting the measured ACS into three ACS aqueous solutions with the concentration from low to high: ACS aqueous solution with first concentration, ACS aqueous solution with second concentration and ACS aqueous solution with third concentration;
in a preferred embodiment, in step S1, the measured ACS is divided into three equal volumes, and then diluted into three different ACS aqueous solutions for use. The mass percentage concentration of the ACS aqueous solution with the first concentration is 4-6%; the mass percentage concentration of the ACS aqueous solution with the second concentration is 8-10%; and the mass percentage concentration of the ACS aqueous solution with the third concentration is 25-35%. More preferably, the ACS aqueous solution of the first concentration is 5% by mass; the mass percentage concentration of the ACS aqueous solution with the second concentration is 10 percent; the ACS aqueous solution with the third concentration has a mass percentage concentration of 30%.
S2, mixing and stirring the measured ACS aqueous solution with the first concentration and 90-95% of chitosan with the high deacetylation degree uniformly to obtain a first mixed solution, and then heating to 50 ℃;
s3, adding 80-85% of measured chitosan with medium deacetylation degree into the heated first mixed solution, uniformly stirring, then starting dropwise adding an ACS aqueous solution with a second concentration, completing dropwise adding within 10 minutes to obtain a second mixed solution, and heating to 80 ℃;
s4, adding metered low-odor fast-volatilization accelerator, uniformly stirring, then beginning to dropwise add ACS aqueous solution with a third concentration, completing dropwise addition within 30 minutes, and reacting for 2-8 hours at 70-90 ℃;
and S5, adding the balance of water, stirring, cooling and cooling to obtain the product. The prepared disinfectant has transparent appearance, and is colorless and tasteless.
As a preferred embodiment, the weight percentage of each metered raw material is as follows: ACS: 1-50%; the sum of the chitosan with medium deacetylation degree and the chitosan with high deacetylation degree is as follows: 30-95%; low odor fast volatilization accelerator: 0.1-5%; water: and (4) the balance.
In a preferred embodiment, the mass ratio of the chitosan with medium deacetylation degree to the chitosan with high deacetylation degree is (1-5): 1.
as a preferred embodiment, the low-odor quick volatilization accelerant is one or a mixture of two of AMP-95 and AMP-96.
The application method of the environment-friendly long-acting no-clean sterilizing disinfectant provided by the invention is to dilute the environment-friendly long-acting no-clean sterilizing disinfectant to 100 times by adding water, and can be used for killing staphylococcus aureus, escherichia coli and/or coronavirus.
The sterilization disinfectant provided by the invention simultaneously adopts the chitosan with the deacetylation degree of 80-85% and the chitosan with the high deacetylation degree of 90-95%, wherein the antibacterial effect of the high deacetylation degree is stronger, the sterilization effect is mainly achieved, the film forming property of the chitosan with the medium deacetylation degree is better, and meanwhile, the sterilization disinfectant has antibacterial ability, and the durability of the sterilization effect can be ensured while the sterilization effect of the product is ensured.
In the preparation process of the sterilizing disinfectant, ACS is diluted into three aqueous solutions with different concentrations and heights, the three aqueous solutions are added for three times, the ACS aqueous solution with lower concentration and weaker acidity is firstly used for assisting the chitosan to be dissolved, the solubility of the chitosan in the weak acid solution is better, the pH value of the ACS aqueous solution with higher concentration and the third concentration is relatively lower and stronger in acidity, and the ACS aqueous solution is added finally, and the low-odor quick-volatilization accelerant is required to be used for firstly adjusting the pH value and then assisting the mixture to be dissolved.
In the preparation method, the production cost and the product quality can be better ensured simultaneously by controlling the reaction time and the temperature of each step.
In order to better illustrate the content of the invention, the invention is further verified by the following specific examples and comparative examples. It should be noted that the examples are given for the purpose of describing the invention more directly and are only a part of the present invention, which should not be construed as limiting the invention in any way.
Composition and preparation of Components of examples and comparative examples
1. The components of each example and comparative example constitute the following table 1:
commercially available finished products were used for each raw material component used in the examples and comparative examples. However, for purposes of the present invention, the components of each raw material are obtained including, but not limited to, those commercially available or prepared by methods provided in the art.
Table 1: the mass percentage of each raw material component of the novel environment-friendly sterilization disinfectant
2. The components measured in Table 1 above were used to prepare a sterilizing disinfectant solution as follows
S1, dividing the measured ACS into three parts with equal volume, and then respectively diluting the three parts into three ACS aqueous solutions with the concentration from low to high: an ACS aqueous solution of a first concentration, the concentration being 5%; an ACS aqueous solution of a second concentration, the concentration being 10%; an ACS aqueous solution of a third concentration, 30%;
s2, mixing and stirring an ACS aqueous solution with the concentration of 5% and metered chitosan with the high deacetylation degree of 90-95% uniformly to obtain a first mixed solution, and then heating to 50 ℃;
s3, adding 80-85% of measured chitosan with medium deacetylation degree into the heated first mixed solution, uniformly stirring, then starting to dropwise add 10% ACS aqueous solution within 10 minutes to obtain a second mixed solution, and heating to 80 ℃;
s4, adding metered AMP-95 and AMP-96, uniformly stirring, then beginning to dropwise add 30% ACS aqueous solution, completing dropwise addition within 30 minutes, and reacting for 2-8 hours at 70-90 ℃;
and S5, adding the balance of water, stirring, cooling and cooling to obtain the product. The prepared disinfectant has transparent appearance, and is colorless and tasteless.
S6, diluting for later use: and (4) mixing the sterilizing disinfectant obtained in the step S5 with disinfectant liquid: water 1: 99 is diluted by adding water into the sterilizing disinfectant to form 100 times of the sterilizing disinfectant for later use.
It is intended to be noted that the preparation method of the present invention is to emphasize that ACS should be added in three times, and the ACS with low concentration and weak acidity should be added in a concentration from low to high, so that chitosan dissolution is facilitated, and then ACS with higher and high concentrations should be added in sequence, and ACS with high concentration should be added with low-odor fast-volatilization promoter to adjust pH value first and then to assist mixing dissolution. Therefore, the present invention only provides the above embodiment of diluting ACS three equal parts to three concentrations of 5%, 10% and 30%, but the present invention is not limited to these three concentrations.
Second, test of sterilizing Performance of sterilizing disinfectant prepared in examples and comparative examples
1. Performance test for killing bacteria
The test methods were described in reference to "Disinfection Specification [ s ] -2002: taking a sterile test tube, adding the test bacteria suspension, adding 0.5mL of 3% bovine serum albumin solution, fully mixing, and preserving the heat in a water bath at 20 ℃ for 5 min. Sucking 4mL of 100 times diluted solution of the sterilizing disinfectant prepared in examples 1-5 and comparative examples 1-2, injecting, acting for 2min and 10min respectively, transferring 0.5mL of the mixed solution into a 4.5mL neutralizing agent test tube, blowing uniformly, acting for 10min, sucking 1mL of the mixed solution, inoculating to a culture dish, placing in a constant-temperature cell incubator, culturing for 48h, and counting viable bacteria. The positive control group is used for replacing the function of a sterilization disinfectant product by using a conventional PBS buffer solution, and the negative control group is used for enabling the sterilization disinfectant to act on a culture medium which is not inoculated with bacteria. Results were averaged over 6 results. The test results are shown in table 2 below.
TABLE 2 results of sterilization Performance test of each example and comparative example
The higher the kill log value, the better the kill effect of the bacteria. The detection result shows that: the 100-time diluent of the sterilization disinfectant provided by the invention has a very good killing effect on escherichia coli and staphylococcus aureus. Comparative example 2 ACS alone is more effective than examples 1-5 in sterilization, but ACS is corrosive and thus cannot be used directly for sterilization of skin, medical devices, and other surfaces. The ACS and the chitosan are synergistic, so that a good sterilization effect can be achieved, and the sterilization composition has the characteristics of no stimulation, no washing, environmental protection and the like.
2. Corona virus killing Performance test
The sterilized disinfectant products prepared in the embodiments 1,4 and 5 are diluted by 100 times for standby, the virus inactivation effect is detected by a third party detection mechanism, the virus detection model is feline coronavirus FCV-WSU79.1683, and the cell model is Hep-G2 cells. The immunofluorescent-oven assay was carried out using commercially available Feline Coronavir (FCoV) Feline fluorescence PCR kit, human serum albumin, fetal bovine serum, and a buffer solution of sodium thiosulfate phosphate containing lecithin (PBS buffer solution). The used instruments are LW-200 fluorescence microscope and CO2Constant temperature incubator according to Disinfection technical Specification (S)]Vaccination with the protocol of-2002, viral Titer (TCID)50) The determination is carried out by adopting a 96-well plate conventional method, the positive control group has the effect of PBS buffer solution replacing a sterilization disinfectant product, and the negative control group does not contain virusIs added directly to the cell culture plate. During detection, each group of experiment plates which are cultured for 48 hours and infected with viruses are taken, culture solution is sucked out and washed by PBS buffer solution for 3 times, then 80% acetone solution is added, the experiment plates are treated at the temperature of-20 ℃ for 20min, the acetone solution is sucked out, the fluorescence kit reagent is completely thawed and then is centrifugally treated, diluted by 100 times by matched diluent, 100ul of diluted reagent solution is added, the reagent solution is placed in a 37 ℃ water bath for 1 hour, the reagent solution is sucked out and washed by PBS for three times, then observation is carried out under an LW-200 fluorescence microscope, the experiment results are the average value of the results of six groups of experiments, and the test results are shown in the following tables 4 and 5.
Viral Titer (TCID)50) The calculation formula of (2) is as follows:
wherein: a is the dilution of greater than 50% cell death, B is greater than 50% cell death, and C is less than 50% cell death.
The inactivation logarithm value is calculated by the following formula: mean log inactivation value ═ logD1-LogD0，
Wherein: d1TCID as positive control group50，D0TCID for Experimental groups50。
The test results are shown in the following tables 3-5:
table 3 example 1 test results of fluorescence detection of inactivated virus
Table 4 example 4 inactivated virus fluorescence detection assay results
TABLE 5 example 5 inactivated Virus fluorescence detection assay results
The results of the positive control group and the experimental group show that: the sterilization disinfectant products of the embodiments 1,4 and 5 of the invention can effectively inactivate feline coronavirus, and the disinfectant of the embodiment 5 has the best virus inactivation effect.
Third, acute oral toxicity test
The test method comprises the following steps: the standard adopted GB 15193.3-2014. The sterilized disinfectant prepared in example 1, example 4, and example 5 was diluted 100 times, and 62.5mL of the diluted solution was taken and put in a 250mL volumetric flask. 8w ICR mice (Schlekschad laboratory animals Co., Ltd. in Hunan) were selected, and 10 mice each were used. The weight of the human body is 18-20 g. The designed amount of the test substance is 5000 mg/kg. Mice were fasted for 16h before gavage, with free access to water. Each mouse was gavaged once every 6h, for a total of two times. Free feeding was restored 3h after gavage, and mice were observed and recorded for 14 consecutive days for performance and mortality. The results of the experiment are shown in table 6 below.
TABLE 6 ICR mouse acute toxicity oral test results
The results show that: the LD50 value of the 100-time diluent of the sterilizing disinfectant is more than 5000mg/kg, and the sterilizing disinfectant belongs to actual nontoxic substances according to acute toxicity classification.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Variations or modifications to the above-described embodiments may be made by those skilled in the art without departing from the spirit and scope of the invention. Therefore, all equivalent modifications or changes that can be made by those skilled in the art without departing from the spirit and technical idea of the present invention shall be covered by the claims of the present invention.
Claims (6)
1. A preparation method of an environment-friendly long-acting washing-free sterilizing disinfectant is characterized by comprising the following steps:
s1, diluting the measured ACS into three ACS aqueous solutions with the concentration from low to high: ACS aqueous solution with first concentration, ACS aqueous solution with second concentration and ACS aqueous solution with third concentration; the mass percentage concentration of the ACS aqueous solution with the first concentration is 4-6%; the mass percentage concentration of the ACS aqueous solution with the second concentration is 8-10%; the mass percentage concentration of the ACS aqueous solution with the third concentration is 25-35%;
s2, mixing and stirring the measured ACS aqueous solution with the first concentration and 90-95% of chitosan with the high deacetylation degree uniformly to obtain a first mixed solution, and then heating to 50 ℃;
s3, adding 80-85% of measured chitosan with medium deacetylation degree into the heated first mixed solution, uniformly stirring, then starting dropwise adding an ACS aqueous solution with a second concentration, completing dropwise adding within 10 minutes to obtain a second mixed solution, and heating to 80 ℃;
s4, adding metered low-odor fast-volatilization accelerator, uniformly stirring, then beginning to dropwise add ACS aqueous solution with a third concentration, completing dropwise addition within 30 minutes, and reacting for 2-8 hours at 70-90 ℃;
s5, adding the balance of water, stirring, cooling and cooling to obtain the environment-friendly long-acting washing-free sterilizing disinfectant;
the method comprises the following raw materials in percentage by mass:
ACS: 1-50%; the sum of the chitosan with medium deacetylation degree and the chitosan with high deacetylation degree is as follows: 30-95%, wherein the mass ratio of the chitosan with medium deacetylation degree to the chitosan with high deacetylation degree is (1-5): 1; low odor fast volatilization accelerator: 0.1-5%; water: and (4) the balance.
2. The production method according to claim 1,
in step S1, the measured ACS is divided into three equal volumes, and then diluted into three ACS aqueous solutions with low to high concentrations: and the ACS aqueous solution with the first concentration, the ACS aqueous solution with the second concentration and the ACS aqueous solution with the third concentration are used for standby.
3. The production method according to claim 1,
the mass percentage concentration of the ACS aqueous solution with the first concentration is 5%; the mass percentage concentration of the ACS aqueous solution with the second concentration is 10 percent; the ACS aqueous solution with the third concentration has a mass percentage concentration of 30%.
4. The production method according to claim 1, 2 or 3,
the low-odor quick-volatilization promoter is one or a mixture of two of AMP-95 and AMP-96.
5. The method of claim 4, wherein the low-odor fast-volatilization promoter is a mixture of two of AMP-95 and AMP-96.
6. The environment-friendly long-acting no-clean sterilizing disinfectant prepared by the method according to any one of claims 1 to 5, which is characterized by comprising the following raw material components in percentage by mass:
ACS：1～50%；
and (3) chitosan: 30-95%;
low odor fast volatilization accelerator: 0.1-5%;
water: the balance;
wherein:
the ACS is added in three ACS aqueous solutions with different mass percentage concentrations, and specifically comprises an ACS aqueous solution with a first concentration, an ACS aqueous solution with a second concentration and an ACS aqueous solution with a third concentration; the ACS aqueous solution with the first concentration has the lowest concentration, and the ACS aqueous solution with the third concentration has the highest concentration; specifically, the mass percentage concentration of the ACS aqueous solution with the first concentration is 4-6%; the mass percentage concentration of the ACS aqueous solution with the second concentration is 8-10%; the mass percentage concentration of the ACS aqueous solution with the third concentration is 25-35%;
the chitosan consists of chitosan with a medium deacetylation degree of 80-85% and chitosan with a high deacetylation degree of 90-95%; and the mass ratio of the chitosan with medium deacetylation degree to the chitosan with high deacetylation degree is (1-5): 1.
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