CN111714455A - Quaternary ammonium salinization chitin anti-pathogenic microorganism spray, preparation method and application thereof - Google Patents
Quaternary ammonium salinization chitin anti-pathogenic microorganism spray, preparation method and application thereof Download PDFInfo
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- CN111714455A CN111714455A CN202010510996.6A CN202010510996A CN111714455A CN 111714455 A CN111714455 A CN 111714455A CN 202010510996 A CN202010510996 A CN 202010510996A CN 111714455 A CN111714455 A CN 111714455A
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- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
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- A—HUMAN NECESSITIES
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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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- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
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- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
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Abstract
The invention provides a salinized quaternary ammonium chitin anti-pathogenic microorganism spray, a preparation method and application thereof, wherein the salinized quaternary ammonium chitin anti-pathogenic microorganism spray comprises the following components: the concentration of the quaternary ammonium salinization chitin is 0.2 to 1.0 weight percent, the concentration of the glycerol is 0.5 to 3.5 weight percent, the concentration of the dexpanthenol is 0.3 to 4.0 weight percent, and the balance is medical sterile pure water; wherein, the deacetylation degree of the quaternary ammonium salinization chitin is 7.4-60.5%, and the substitution degree of the quaternary ammonium salt is 0.28-0.82. The quaternary ammonium salinization chitin anti-pathogenic microorganism spray provided by the invention has the characteristics of excellent biological safety, no toxicity, no immunogenicity and biodegradability, and has various effects including broad-spectrum efficient antibacterial and antiviral properties and promotion of wound healing, and the preparation method is simple and practical and is beneficial to industrial production.
Description
Technical Field
The invention belongs to the field of biomedical materials, and particularly relates to a quaternary ammonium salinization chitin anti-pathogenic microorganism spray, a preparation method and application thereof.
Background
Chitin is a crystalline polysaccharide composed of beta- (1 → 4) -2-acetamido-D-glucose, widely exists in exoskeletons of aquatic animals such as shrimps and crabs, the horny layer of invertebrates, green algae, fungi and yeast cell walls, and is a natural polymer with the next accumulation amount to cellulose on the earth. The chitosan is a product obtained by deacetylation reaction of chitin, so that the solubility is further improved, and the amino group with positive charges on the surface also enhances the antibacterial performance. The chitin and the chitosan are derived from organisms, are renewable natural polymer materials, have the characteristics of good biocompatibility, biodegradability, no toxicity, no immunogenicity and the like, and are widely applied to the fields of research and product development of biomedical materials such as antibiosis, hemostasis, wound healing promotion, antivirus and the like. However, the most common method for preparing chitosan is to deacetylate chitin under the conditions of concentrated alkali and high temperature, which has high cost and low efficiency and causes environmental pollution. And chitin is more stable than chitosan and has natural biocompatibility, so that the chitin is directly used as a matrix for chemical modification and transformation to prepare the multifunctional biomedical material, and the multifunctional biomedical material has higher application value.
Due to the interaction of ultra-strong hydrogen bonds in molecules and among molecules, chitin forms a microfiber-net-shaped highly-crystallized ordered structure, and the chitin is difficult to be directly utilized and still cannot meet the requirements of practical application to a certain extent because the chitin is difficult to dissolve in water, dilute acid, dilute alkali and common organic solvents. The quaternary ammonium salt modification of chitin is an important direction for chemical modification research of chitin, and on the basis of retaining excellent biological safety of chitin, the introduction of quaternary ammonium salt groups greatly improves the solubility of chitin and further enhances the antibacterial performance of chitin. The amino group and the quaternary ammonium salt group with strong positive charges in the quaternary ammonium salinization chitin molecule can be combined with the negative charge component on the pathogenic microorganism biomembrane, so that the structure and the function of the microorganism membrane are damaged, and the microorganism membrane has the efficient and broad-spectrum pathogenic microorganism resistance effect; meanwhile, the quaternary ammonium salinization chitin can also be used for wound surface repair, can reduce inflammatory cell infiltration, increases the production of new blood vessels and collagen fibers, and has the functions of resisting infection and promoting healing in vivo. In the field of biomedical material research and development, quaternary ammonium salinized chitin has great potential.
At present, the chitin-based antibacterial material mainly comprises physical forms such as non-woven fabric dressing, hydrogel, powder and coating, and can only be applied to treatment and nursing of patients with clinically damaged wound surfaces, surgical operations and the like, and the application scene is limited. In some special but common application scenes, the chitin-based antibacterial material with excellent performance cannot be practically applied. The mask and the protective clothing have no capability of killing pathogenic microorganisms such as bacteria and viruses, the pathogenic microorganisms remained on the waste mask and the protective clothing are very important potential infection sources, and the waste mask and the protective clothing are discarded at will without medical treatment, so that serious secondary pollution is easily caused. At the same time, a large number of medical devices also require treatment with anti-pathogenic microbial materials to block nosocomial infections. If the quaternary ammonium salinization chitin can be made into spray in a certain way and sprayed on the surfaces of masks, protective clothing and related medical appliances, the static action generated by amino and quaternary ammonium salt molecules in the quaternary ammonium salinization chitin can be utilized to adsorb and kill residual pathogenic microorganisms such as bacteria and viruses, and the secondary pollution is blocked. The quaternary ammonium salinization chitin spraying agent can be applied to treatment and nursing of wound patients, and the spraying agent can be contacted with the wound more uniformly and fully, so that the effects of resisting infection and promoting wound healing are achieved.
Therefore, the quaternary ammonium salinization chitin spraying agent with broad-spectrum efficient anti-pathogenic microorganism performance and wound healing promotion performance is developed, an efficient preparation method is searched, and the quaternary ammonium salinization chitin spraying agent has a good application prospect.
Disclosure of Invention
The invention aims to provide a quaternary ammonium salinization chitin anti-pathogenic microorganism spray which has the characteristics of excellent biological safety, no toxicity, no immunogenicity and biodegradability, and has multiple functions of broad-spectrum efficient antibiosis, antivirus performance and wound healing promotion.
The invention also aims to provide a preparation method of the quaternary ammonium salinization chitin anti-pathogenic microorganism spray, which is simple and practical and is beneficial to industrial production.
The invention also aims to provide application of the quaternary ammonium salinization chitin anti-pathogenic microorganism spray in antibiosis, antivirus and promotion of wound healing.
One of the purposes of the invention adopts the following technical scheme:
a quaternary ammonium salinization chitin anti-pathogenic microorganism spray comprises the following components:
the concentration of the quaternary ammonium salinization chitin is 0.2 to 1.0 weight percent, the concentration of the glycerol is 0.5 to 3.5 weight percent, the concentration of the dexpanthenol is 0.3 to 4.0 weight percent, and the balance is medical sterile pure water; wherein, the deacetylation degree of the quaternary ammonium salinization chitin is 7.4-60.5%, and the substitution degree of the quaternary ammonium salt is 0.28-0.82.
Preferably, the concentration of the quaternary ammonium salinization chitin is 0.8 wt%, the concentration of glycerol is 1.5 wt%, the concentration of dexpanthenol is 3.5 wt%, and the balance is medical sterile pure water, the deacetylation degree of the quaternary ammonium salinization chitin is 45.8-55.3%, and the substitution degree of quaternary ammonium salt is 0.46-0.82.
The second purpose of the invention is realized by adopting the following technical scheme:
a preparation method of a quaternary ammonium salinization chitin anti-pathogenic microorganism spray comprises the following steps:
(1) preparing raw materials according to the following mass percentages:
the concentration of the quaternary ammonium salinization chitin is 0.2 to 1.0 weight percent, the concentration of the glycerol is 0.5 to 3.5 weight percent, the concentration of the dexpanthenol is 0.3 to 4.0 weight percent, and the balance is medical sterile pure water; wherein, the deacetylation degree of the quaternary ammonium salinization chitin is 7.4-60.5%, and the substitution degree of the quaternary ammonium salt is 0.28-0.82;
(2) firstly, adding medical sterile pure water into a cleaned and dried stirrer;
(3) adding the quaternary ammonium salinization chitin into the stirrer in the step (2), and continuously stirring until the quaternary ammonium salinization chitin is fully dissolved to obtain a quaternary ammonium salinization chitin solution;
(4) filtering to remove solid residual impurities in the quaternary ammonium salinization chitin solution in the step (3), and dialyzing with sterile pure water until chloride ions are completely removed;
(5) continuously stirring the quaternary ammonium salinization chitin solution obtained after dialysis in the step (4), slowly adding the glycerol, and stirring for 10-20 minutes until the solution is uniformly mixed;
(6) adding the dexpanthenol into the uniformly mixed solution obtained in the step (5), and continuously stirring until the mixture is completely and uniformly mixed to obtain a quaternary ammonium salinization chitin anti-pathogenic microorganism spray solution;
(7) and (4) after the quaternary ammonium salinization chitin anti-pathogenic microorganism spray solution obtained in the step (6) is detected to meet the product standard, packaging by using a medical-grade spray bottle to obtain a finished product.
Further, in the step (4), the filtering is specifically to select a polycarbonate microporous filter membrane with the pore size of 5 μm, and filter to remove solid residual impurities in the solution; specifically, the dialysis is carried out by selecting a MW 8000-14000 dialysis bag, dialyzing the quaternary ammonium salt possibly remaining in the sterile pure water environment, and determining the concentration of chloride ions in the dialysate by adopting a silver ion titration method until the chloride ions are completely removed, thus obtaining the dialyzed quaternary ammonium salinized chitin solution.
The third technical scheme for realizing the purpose of the invention is as follows:
an application of the quaternary ammonium salinization chitin anti-pathogenic microorganism spray in resisting bacteria and virus and promoting wound healing is provided.
Further, the fungi include Escherichia coli, Staphylococcus aureus, Candida albicans, MRSA, Acinetobacter baumannii, Bacteroides fragilis, and Klebsiella pneumoniae.
Further, the virus includes a novel coronavirus.
Compared with the prior art, the invention has the following beneficial effects:
(1) the quaternary ammonium salinization chitin applied in the technical scheme of the invention has the characteristics of good biological safety, biodegradability, no toxicity, no immunogenicity and the like, and simultaneously has the pathogenic microorganism performances of broad-spectrum efficient antibiosis, antivirus and the like and the wound healing promotion performance. In addition, the low-concentration glycerin has good moisture retention, can keep the wound moist, accords with the treatment theory of moist healing of the wound surface, and can also play a role in protecting the skin and the mucosa by adding the dexpanthenol.
(2) The quaternary ammonium salinization chitin anti-pathogenic microorganism spraying agent provided by the invention has the anti-pathogenic microorganism performance stronger than that of common antibacterial spraying agents such as chitosan, and the main antibacterial mechanism is that amino groups and quaternary ammonium salt groups with strong positive charges in quaternary ammonium salinization chitin molecules can be combined with negative charge components on a pathogenic microorganism biological membrane to cause the structure and the function of the microbial membrane to be damaged, so that pathogenic microorganisms are killed. The quaternary ammonium salinization chitin is prepared into the spray for the first time, the spray is not only used for treating common acute wound surface, infective wound surface and chronic wound surface infection difficult to heal clinically and accelerating wound healing, but also can be sprayed on the surfaces of waste masks, protective clothing and related medical appliances at any time and any place when epidemic situations such as influenza, new crown pneumonia and the like occur, so that pathogenic microorganisms such as residual bacteria, viruses and the like are killed, the propagation path is cut off, the risk of secondary pollution is reduced, and the diffusion of infection is effectively restrained.
(3) The quaternary ammonium salinization chitin anti-pathogenic microorganism spray provided by the invention can resist escherichia coli, staphylococcus aureus, candida albicans, MRSA, multi-resistant acinetobacter baumannii, bacteroides fragilis and klebsiella pneumoniae, has a virus inhibition rate of more than 95% on novel coronavirus, has a good effect of resisting the novel coronavirus, and has important significance for further research and use of the quaternary ammonium salinization chitin anti-pathogenic microorganism spray.
(4) The quaternary ammonium salinization chitin anti-pathogenic microorganism spray provided by the invention is convenient to use and very wide in application, and can be used for treating skin abrasion, living goods possibly polluted by pathogenic microorganisms and the like. Meanwhile, the quaternary ammonium salinization chitin anti-pathogenic microorganism spray in the technical scheme has the advantages of wide sources of preparation raw materials, low cost, simple and practical preparation method, contribution to industrial production and wide application prospect.
Detailed Description
The features and advantages of the present invention will be further understood by the following detailed description. The examples provided are merely illustrative of the method of the present invention and do not limit the remainder of the disclosure in any way.
The experimental methods used in the following examples of the present invention are all conventional methods unless otherwise specified. The raw materials of the medical quaternary ammonium salinization chitin, glycerin, dexpanthenol and medical sterile pure water used in the following examples are all commercially available medical products. The materials, reagents and the like used are commercially available unless otherwise specified.
[ example 1 ]
Preparing the quaternary ammonium salinization chitin anti-pathogenic microorganism spray according to the following steps:
(1) preparing raw materials according to the following components: 20g of quaternary ammonium salinization chitin; 350g of glycerol; dexpanthenol is 30 g;
(2) firstly, 8L of medical sterile pure water is added into a stirrer after being cleaned and dried;
(3) adding the quaternary ammonium salinized chitin into a stirrer, and continuously stirring for 1 hour until the quaternary ammonium salinized chitin is fully dissolved;
(4) filtering to remove solid residual impurities in the solution, and dialyzing with sterile pure water until chloride ions are completely removed;
(5) continuously stirring the completely dissolved quaternary ammonium salinization chitin solution obtained after dialysis and slowly adding glycerin, and stirring for 10-20 minutes until the solution is uniformly mixed;
(6) adding dexpanthenol, then fixing the volume to 10L, and continuously stirring until the mixture is completely and uniformly mixed;
(7) and (5) conveying to the next procedure for subpackage, and packaging by using a medical grade spray bottle after the detection that the product meets the standard to obtain a finished product.
In this example, the deacetylation degree of the quaternized chitin was 7.4% and the substitution degree of the quaternary ammonium salt was 0.28.
In this embodiment, before preparing the quaternary ammonium salinized chitin anti-pathogenic microorganism spray, the selected quaternary ammonium salinized chitin should be subjected to purification dialysis, and the purification dialysis specifically comprises the following steps: selecting a polycarbonate microporous filter membrane with the aperture of 5 mu m, and filtering to remove solid residual impurities in the solution; then dialyzing possible residual quaternary ammonium salt in a sterile pure water environment by using a MW 8000-14000 dialysis bag, and determining the concentration of chloride ions in the dialyzate by adopting a silver ion titration method until the chloride ions are completely removed to obtain the quaternary ammonium salinized chitin solution which can be used subsequently.
[ example 2 ]
Preparing the quaternary ammonium salinization chitin anti-pathogenic microorganism spray according to the following steps:
(1) preparing raw materials according to the following components: 50g of quaternary ammonium salinization chitin; 200g of glycerol; dexpanthenol is 150 g;
(2) firstly, 8L of medical sterile pure water is added into a stirrer after being cleaned and dried;
(3) adding the quaternary ammonium salinized chitin into a stirrer, and continuously stirring for 1 hour until the quaternary ammonium salinized chitin is fully dissolved;
(4) filtering to remove solid residual impurities in the solution, and dialyzing with sterile pure water until chloride ions are completely removed;
(5) continuously stirring the completely dissolved quaternary ammonium salinization chitin solution obtained after dialysis and slowly adding glycerin, and stirring for 10-20 minutes until the solution is uniformly mixed;
(6) adding dexpanthenol, then fixing the volume to 10L, and continuously stirring until the mixture is completely and uniformly mixed;
(7) and (5) conveying to the next procedure for subpackage, and packaging by using a medical grade spray bottle after the detection that the product meets the standard to obtain a finished product.
In this example, the deacetylation degree of the quaternized chitin was 45.8% and the substitution degree of the quaternary ammonium salt was 0.82.
In this embodiment, before preparing the quaternary ammonium salinized chitin anti-pathogenic microorganism spray, the selected quaternary ammonium salinized chitin should be subjected to purification dialysis, and the purification dialysis specifically comprises the following steps: selecting a polycarbonate microporous filter membrane with the aperture of 5 mu m, and filtering to remove solid residual impurities in the solution; then dialyzing possible residual quaternary ammonium salt in a sterile pure water environment by using a MW 8000-14000 dialysis bag, and determining the concentration of chloride ions in the dialyzate by adopting a silver ion titration method until the chloride ions are completely removed to obtain the quaternary ammonium salinized chitin solution which can be used subsequently.
[ example 3 ]
Preparing the quaternary ammonium salinization chitin anti-pathogenic microorganism spray according to the following steps:
(1) preparing raw materials according to the following components: 80g of quaternary ammonium salinization chitin; 150g of glycerol; dexpanthenol is 350 g;
(2) firstly, 8L of medical sterile pure water is added into a stirrer after being cleaned and dried;
(3) adding the quaternary ammonium salinized chitin into a stirrer, and continuously stirring for 1 hour until the quaternary ammonium salinized chitin is fully dissolved;
(4) filtering to remove solid residual impurities in the solution, and dialyzing with sterile pure water until chloride ions are completely removed;
(5) continuously stirring the completely dissolved quaternary ammonium salinization chitin solution obtained after dialysis and slowly adding glycerin, and stirring for 10-20 minutes until the solution is uniformly mixed;
(6) adding dexpanthenol, then fixing the volume to 10L, and continuously stirring until the mixture is completely and uniformly mixed;
(7) and (5) conveying to the next procedure for subpackage, and packaging by using a medical grade spray bottle after the detection that the product meets the standard to obtain a finished product.
In this example, the deacetylation degree of the quaternized chitin was 49.1%, and the substitution degree of the quaternary ammonium salt was 0.61.
In this embodiment, before preparing the quaternary ammonium salinized chitin anti-pathogenic microorganism spray, the selected quaternary ammonium salinized chitin should be subjected to purification dialysis, and the purification dialysis specifically comprises the following steps: selecting a polycarbonate microporous filter membrane with the aperture of 5 mu m, and filtering to remove solid residual impurities in the solution; then dialyzing possible residual quaternary ammonium salt in a sterile pure water environment by using a MW 8000-14000 dialysis bag, and determining the concentration of chloride ions in the dialyzate by adopting a silver ion titration method until the chloride ions are completely removed to obtain the quaternary ammonium salinized chitin solution which can be used subsequently.
[ example 4 ]
Preparing the quaternary ammonium salinization chitin anti-pathogenic microorganism spray according to the following steps:
(1) preparing raw materials according to the following components: 80g of quaternary ammonium salinization chitin; 150g of glycerol; dexpanthenol is 350 g;
(2) firstly, 8L of medical sterile pure water is added into a stirrer after being cleaned and dried;
(3) adding the quaternary ammonium salinized chitin into a stirrer, and continuously stirring for 1 hour until the quaternary ammonium salinized chitin is fully dissolved;
(4) filtering to remove solid residual impurities in the solution, and dialyzing with sterile pure water until chloride ions are completely removed;
(5) continuously stirring the completely dissolved quaternary ammonium salinization chitin solution obtained after dialysis and slowly adding glycerin, and stirring for 10-20 minutes until the solution is uniformly mixed;
(6) adding dexpanthenol, then fixing the volume to 10L, and continuously stirring until the mixture is completely and uniformly mixed;
(7) and (5) conveying to the next procedure for subpackage, and packaging by using a medical grade spray bottle after the detection that the product meets the standard to obtain a finished product.
In this example, the deacetylation degree of the quaternized chitin was 55.3% and the substitution degree of the quaternary ammonium salt was 0.46.
In this embodiment, before preparing the quaternary ammonium salinized chitin anti-pathogenic microorganism spray, the selected quaternary ammonium salinized chitin should be subjected to purification dialysis, and the purification dialysis specifically comprises the following steps: selecting a polycarbonate microporous filter membrane with the aperture of 5 mu m, and filtering to remove solid residual impurities in the solution; then dialyzing possible residual quaternary ammonium salt in a sterile pure water environment by using a MW 8000-14000 dialysis bag, and determining the concentration of chloride ions in the dialyzate by adopting a silver ion titration method until the chloride ions are completely removed to obtain the quaternary ammonium salinized chitin solution which can be used subsequently.
[ example 5 ]
Preparing the quaternary ammonium salinization chitin anti-pathogenic microorganism spray according to the following steps:
(1) preparing raw materials according to the following components: 100g of quaternized chitin; 50g of glycerol; dexpanthenol is 400 g;
(2) firstly, 8L of medical sterile pure water is added into a stirrer after being cleaned and dried;
(3) adding the quaternary ammonium salinized chitin into a stirrer, and continuously stirring for 1 hour until the quaternary ammonium salinized chitin is fully dissolved;
(4) filtering to remove solid residual impurities in the solution, and dialyzing with sterile pure water until chloride ions are completely removed;
(5) continuously stirring the completely dissolved quaternary ammonium salinization chitin solution obtained after dialysis and slowly adding glycerin, and stirring for 10-20 minutes until the solution is uniformly mixed;
(6) adding dexpanthenol, then fixing the volume to 10L, and continuously stirring until the mixture is completely and uniformly mixed;
(7) and (5) conveying to the next procedure for subpackage, and packaging by using a medical grade spray bottle after the detection that the product meets the standard to obtain a finished product.
In this example, the deacetylation degree of the quaternized chitin was 60.5% and the substitution degree of the quaternary ammonium salt was 0.29.
In this embodiment, before preparing the quaternary ammonium salinized chitin anti-pathogenic microorganism spray, the selected quaternary ammonium salinized chitin should be subjected to purification dialysis, and the purification dialysis specifically comprises the following steps: selecting a polycarbonate microporous filter membrane with the aperture of 5 mu m, and filtering to remove solid residual impurities in the solution; then dialyzing possible residual quaternary ammonium salt in a sterile pure water environment by using a MW 8000-14000 dialysis bag, and determining the concentration of chloride ions in the dialyzate by adopting a silver ion titration method until the chloride ions are completely removed to obtain the quaternary ammonium salinized chitin solution which can be used subsequently.
The performance test of the finished product of the quaternary ammonium salinization chitin anti-pathogenic microorganism spray prepared in each embodiment 1-5 is as follows:
(1) testing the in vitro broad-spectrum antibacterial performance of the quaternary ammonium salinization chitin spray:
referring to the provisions of GB15979-2002 hygienic Standard for Disposable sanitary products, the quaternary ammonium salinized chitin spray is tested in vitro for the antibacterial effect on gram-negative bacteria, gram-positive bacteria, fungi, drug-resistant bacteria and anaerobic bacteria, and the experimental results are shown in the following table 1:
TABLE 1
Aiming at the quaternary ammonium salinization chitin spraying agent prepared in the 5 embodiments, 3 samples in different batches are distributed and selected for carrying out antibacterial performance detection, the antibacterial rate of all the samples exceeds 90%, and the quality requirement is met. In addition, as can be seen from table 1, all of the embodiments of example 4 have the best antibacterial effect.
(2) In-vitro antiviral performance test of the quaternary ammonium salinization chitin spray:
50 μ L of 100TCID was added to vero-E6 cells grown in 96-well plates50The novel coronavirus diluent of (1) was adsorbed in a carbon dioxide incubator at 33 ℃ for 1 hour, and the virus solution was discarded and washed 3 times with the virus diluent. The quaternary ammonium salinization chitin spray solution prepared in the example 1-5 is added into the cells which grow into a monolayer in a 96-well plate, 100 mu L of the quaternary ammonium salinization chitin spray solution is added into each well, each concentration is carried out for 4 multiple wells, the culture is carried out at 33 ℃, the CPE result is observed and recorded day by day, and the observation is continuously carried out for 3 days.
Aiming at the quaternary ammonium salinization chitin spraying agent prepared in the 5 embodiments, 3 spraying agent samples in different batches are distributed and selected for antiviral performance test, and experimental results show that the virus inhibition rate of the quaternary ammonium salinization chitin spraying agent obtained in each embodiment on novel coronavirus is over 95%, the effective virus killing time of all samples is within 30min, and the antiviral performance is excellent.
(3) Testing the in vivo anti-infection and wound healing promoting performance of the quaternary ammonium salinization chitin spray:
after anesthesia of SD male rats, hair on the back was shaved with a razor, followed by thorough hair removal treatment with 7% sodium sulfide, 3 minutes later, the skin on the back of the rats was wiped off with physiological saline, after the skin on the back was sterilized, skin perforation was performed on both sides of the spinal column on the back of the rats with a 1.5 cm-diameter punch until the muscle layer was destroyed, and 100. mu.l of 1 × 10 was dropped onto the wound site8Forming an infected wound model after 48 hours by using the CFU/ml staphylococcus aureus liquid; the blank group covered the wound surface with common medical gauze, and the experimental group evenly sprayed the quaternary ammonium salinization chitin spraying agent prepared in the above examples 1-5 on the wound surface respectively, wherein the spraying time lasts more than 30 seconds; after the spraying agent acts for 3min, the wound is properly sealed by using the air-permeable remaining needle film and the medical adhesive tape, and the wound infection and healing days are dynamically observed.
The experimental results show that the average healing time of the infective wounds after the treatment of the quaternary ammonium salt chitin spray obtained in the example 1 is 23 +/-1 days, the average healing time of the infective wounds after the treatment of the quaternary ammonium salt chitin spray obtained in the example 2 is 23 +/-1 days, the average healing time of the infective wounds after the treatment of the quaternary ammonium salt chitin spray obtained in the example 3 is 21 +/-1 days, the average healing time of the infective wounds after the treatment of the quaternary ammonium salt chitin spray obtained in the example 4 is 21 +/-1 days, the average healing time of the infective wounds after the treatment of the quaternary ammonium salt chitin spray obtained in the example 5 is 21 +/-1 days, and the average healing time of the infective wounds after the treatment of the blank gauze is 29 +/-1 days. According to statistical analysis, the quaternary ammonium salinization chitin spraying agent has the obvious performance of promoting the healing of infectious wound, and compared with common medical gauze, the average healing time is shortened by 20-30%.
(4) Detecting pathogenic microorganisms on the surface of the waste mask treated by the quaternary ammonium salinization chitin spray:
the quats prepared in examples 1-5 above were combinedSpraying the spray with salted chitin on the outer surface of medical protective mask, controlling the spraying thickness to be 1-2mm, standing for 10-15min, cutting the mask fabric into small pieces with square shape of 2cm × 2cm, and respectively collecting 100 μ L of cultured golden yellow staphylococcus liquid, Escherichia coli liquid and Klebsiella pneumoniae liquid (the number of bacteria is about 10)6cfu/mL) were dropped onto the surface of different mask cubes and spread evenly with a sterile spatula. Placing the small square mask with the bacteria liquid in a sterile bacteria culture dish, placing at room temperature in sterile environment for 12h, placing the small square mask in a sterile centrifuge tube preheated at 37 ℃ and containing 5mL of LB bacterial culture medium, removing the small square mask after shaking and swirling for 1min, centrifuging, removing supernatant, and diluting with LB bacterial culture medium by 0 time, 10 times respectively210 times of310 times of4After doubling, 100. mu.L of each was spread on LB solid plate medium, cultured at 37 ℃ for 24 hours, and checked for residual bacterial survival. Residual virus survival was determined similarly to the above method. Take 100. mu.L of 100TCID50The novel coronavirus diluent is dripped on the surfaces of different mask small square pieces and is evenly smeared by a sterile tongue depressor. Placing the small square mask dropwise added with the virus diluent into a sterile bacteria culture dish, placing the small square mask in a sterile environment at room temperature for 12 hours, placing the small square mask into a sterile centrifuge tube precooled at 4 ℃ and containing 5mL of virus transport leaves, removing the small square mask after shaking and vortexing for 1min, removing supernatant after centrifugation at 4 ℃, performing suspension precipitation by using 1mL of culture solution, performing cryopreservation at-80 ℃, freezing and thawing before use, adding a double antibody, and performing action at 4 ℃ for 1-2 hours to obtain the inoculum. Vero-E6 cells overgrow 80-90% within 48h of passage, inoculated with virus inoculation liquid, and treated in the same way by replacing inoculum with maintenance liquid, and used as normal cell control. The bottles were examined daily from the next day for the presence of CPE cytopathic effect typical of the novel coronavirus until day 5.
Aiming at the quaternary ammonium salinization chitin spraying agent prepared in the 5 embodiments, 3 mask samples treated by spraying agents in different batches are distributed and selected for detection of pathogenic microorganisms, and experimental results show that the protective mask treated by the quaternary ammonium salinization chitin spraying agent obtained in each embodiment has the bacteriostasis rate of over 99 percent on staphylococcus aureus, escherichia coli and klebsiella pneumoniae, and novel coronavirus remained on the surface of the mask treated by the spraying agent is dead and does not survive.
The foregoing is a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications and variations can be made without departing from the technical principle of the present invention, and these modifications and variations should also be regarded as the protection scope of the present invention.
Claims (7)
1. The quaternary ammonium salinization chitin anti-pathogenic microorganism spray is characterized in that: comprises the following components:
the concentration of the quaternary ammonium salinization chitin is 0.2 to 1.0 weight percent, the concentration of the glycerol is 0.5 to 3.5 weight percent, the concentration of the dexpanthenol is 0.3 to 4.0 weight percent, and the balance is medical sterile pure water; wherein, the deacetylation degree of the quaternary ammonium salinization chitin is 7.4-60.5%, and the substitution degree of the quaternary ammonium salt is 0.28-0.82.
2. The quaternized chitin pathogenic microorganism spray of claim 1, wherein: the quaternary ammonium salinization chitin is 0.8 wt%, the glycerol is 1.5 wt%, the dexpanthenol is 3.5 wt%, and the balance is medical sterile pure water, wherein the deacetylation degree of the quaternary ammonium salinization chitin is 45.8-55.3%, and the substitution degree of the quaternary ammonium salt is 0.46-0.82.
3. A method of preparing the quaternized chitin anti-pathogenic microorganism spray of claim 1 or 2, characterized in that: the method comprises the following steps:
(1) preparing raw materials according to the following mass percentages:
the concentration of the quaternary ammonium salinization chitin is 0.2 to 1.0 weight percent, the concentration of the glycerol is 0.5 to 3.5 weight percent, the concentration of the dexpanthenol is 0.3 to 4.0 weight percent, and the balance is medical sterile pure water; wherein, the deacetylation degree of the quaternary ammonium salinization chitin is 7.4-60.5%, and the substitution degree of the quaternary ammonium salt is 0.28-0.82;
(2) firstly, adding medical sterile pure water into a cleaned and dried stirrer;
(3) adding the quaternary ammonium salinization chitin into the stirrer in the step (2), and continuously stirring until the quaternary ammonium salinization chitin is fully dissolved to obtain a quaternary ammonium salinization chitin solution;
(4) filtering to remove solid residual impurities in the quaternary ammonium salinization chitin solution in the step (3), and dialyzing with sterile pure water until chloride ions are completely removed;
(5) continuously stirring the quaternary ammonium salinization chitin solution obtained after dialysis in the step (4), slowly adding the glycerol, and stirring for 10-20 minutes until the solution is uniformly mixed;
(6) adding the dexpanthenol into the uniformly mixed solution obtained in the step (5), and continuously stirring until the mixture is completely and uniformly mixed to obtain a quaternary ammonium salinization chitin anti-pathogenic microorganism spray solution;
(7) and (4) after the quaternary ammonium salinization chitin anti-pathogenic microorganism spray solution obtained in the step (6) is detected to meet the product standard, packaging by using a medical-grade spray bottle to obtain a finished product.
4. The method of making the quaternized chitin pathogenic microorganism spray of claim 3, wherein: in the step (4), the filtration is specifically to select a polycarbonate microporous filter membrane with the pore size of 5 microns, and filter to remove solid residual impurities in the solution; specifically, the dialysis is carried out by selecting a MW 8000-14000 dialysis bag, dialyzing the quaternary ammonium salt possibly remaining in the sterile pure water environment, and determining the concentration of chloride ions in the dialysate by adopting a silver ion titration method until the chloride ions are completely removed, thus obtaining the dialyzed quaternary ammonium salinized chitin solution.
5. Use of the quaternized chitin anti-pathogenic microorganism spray of claim 1 for antibacterial, antiviral and wound healing promotion.
6. The use of claim 5, wherein: the bacteria include Escherichia coli, Staphylococcus aureus, Candida albicans, MRSA, Acinetobacter baumannii, Bacteroides fragilis, and Klebsiella pneumoniae.
7. The use of claim 5, wherein: the viruses include novel coronaviruses.
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| CN112662562A (en) * | 2021-01-12 | 2021-04-16 | 哈尔滨工业大学 | Method for targeted capture and killing of escherichia coli by constructing artificial cell model based on yeast cell wall camouflage |
| CN112662562B (en) * | 2021-01-12 | 2023-01-06 | 哈尔滨工业大学 | Method for targeted capture and killing of escherichia coli by constructing artificial cell model based on yeast cell wall camouflage |
| CN116041568A (en) * | 2023-01-11 | 2023-05-02 | 武汉大学中南医院 | Water-soluble imidazole salt chitin with broad-spectrum antibacterial property, preparation method and application thereof |
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