CN111840211A - Antibacterial gel containing amphoteric alkaloid and preparation method thereof - Google Patents
Antibacterial gel containing amphoteric alkaloid and preparation method thereof Download PDFInfo
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- CN111840211A CN111840211A CN201910345500.1A CN201910345500A CN111840211A CN 111840211 A CN111840211 A CN 111840211A CN 201910345500 A CN201910345500 A CN 201910345500A CN 111840211 A CN111840211 A CN 111840211A
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- 229930013930 alkaloid Natural products 0.000 title claims abstract description 44
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 38
- 150000003797 alkaloid derivatives Chemical class 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 239000000661 sodium alginate Substances 0.000 claims abstract description 45
- 235000010413 sodium alginate Nutrition 0.000 claims abstract description 45
- 229940005550 sodium alginate Drugs 0.000 claims abstract description 45
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims abstract description 40
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 33
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 29
- NIXOWILDQLNWCW-UHFFFAOYSA-N Acrylic acid Chemical compound OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims abstract description 27
- 229920002125 Sokalan® Polymers 0.000 claims abstract description 27
- 229960001631 carbomer Drugs 0.000 claims abstract description 27
- 239000008213 purified water Substances 0.000 claims abstract description 26
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 claims abstract description 15
- 239000002994 raw material Substances 0.000 claims abstract description 6
- 238000003756 stirring Methods 0.000 claims description 25
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 16
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 15
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 15
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 11
- -1 nitrogen-containing organic compounds Chemical class 0.000 claims description 11
- JQWHASGSAFIOCM-UHFFFAOYSA-M sodium periodate Chemical compound [Na+].[O-]I(=O)(=O)=O JQWHASGSAFIOCM-UHFFFAOYSA-M 0.000 claims description 10
- 238000000967 suction filtration Methods 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 6
- 238000005303 weighing Methods 0.000 claims description 6
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 5
- 238000004108 freeze drying Methods 0.000 claims description 5
- 230000001590 oxidative effect Effects 0.000 claims description 5
- 230000001376 precipitating effect Effects 0.000 claims description 5
- 239000011780 sodium chloride Substances 0.000 claims description 5
- 238000003892 spreading Methods 0.000 claims description 5
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- 238000000034 method Methods 0.000 claims description 2
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- 206010052428 Wound Diseases 0.000 abstract description 17
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- 206010048038 Wound infection Diseases 0.000 abstract description 5
- 239000000499 gel Substances 0.000 description 38
- 239000000243 solution Substances 0.000 description 33
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- 239000008055 phosphate buffer solution Substances 0.000 description 5
- 125000003277 amino group Chemical group 0.000 description 4
- 239000013642 negative control Substances 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 241000222122 Candida albicans Species 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
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- 206010061218 Inflammation Diseases 0.000 description 3
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- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 3
- 238000009987 spinning Methods 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 235000009161 Espostoa lanata Nutrition 0.000 description 2
- 240000001624 Espostoa lanata Species 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- 150000004753 Schiff bases Chemical class 0.000 description 2
- 125000003172 aldehyde group Chemical group 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
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- 230000005764 inhibitory process Effects 0.000 description 2
- 230000009878 intermolecular interaction Effects 0.000 description 2
- 150000003951 lactams Chemical group 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
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- 239000002674 ointment Substances 0.000 description 2
- 230000002980 postoperative effect Effects 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
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- 206010002091 Anaesthesia Diseases 0.000 description 1
- 206010048946 Anal abscess Diseases 0.000 description 1
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- 206010030113 Oedema Diseases 0.000 description 1
- 208000004680 Rectal Fistula Diseases 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 206010002156 anal fistula Diseases 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
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- 230000000249 desinfective effect Effects 0.000 description 1
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- 210000003195 fascia Anatomy 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 208000035474 group of disease Diseases 0.000 description 1
- 210000004209 hair Anatomy 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 208000035861 hematochezia Diseases 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 230000001050 lubricating effect Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 230000000474 nursing effect Effects 0.000 description 1
- 239000006916 nutrient agar Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000036407 pain Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 125000001453 quaternary ammonium group Chemical group 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 238000007390 skin biopsy Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 238000004381 surface treatment Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0014—Skin, i.e. galenical aspects of topical compositions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Inorganic Chemistry (AREA)
- Dermatology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses an antibacterial gel containing amphoteric alkaloids and a preparation method thereof, wherein the antibacterial gel is prepared from the following raw materials in parts by weight: 1-3 parts of amphoteric alkaloid, 1-3 parts of oxidized sodium alginate, 0.5-2 parts of carbomer, 0.5-4 parts of triethanolamine, 7-13 parts of glycerol and 100 parts of purified water. The invention is applied to the protection of anorectal diseases and perioperative wound care, more effectively prevents wound infection and promotes wound healing.
Description
Technical Field
The invention relates to the technical field of medical instruments, in particular to an antibacterial gel containing amphoteric alkaloids and a preparation method thereof.
Background
Anorectal diseases refer to a group of diseases occurring in rectum, anus and surrounding tissues, the types of the diseases are more than 30, and the most common anorectal diseases in clinic comprise four diseases of hemorrhoids, anal fissure, anal fistula and perianal abscess. The anorectal diseases are common and frequent diseases of human beings, have high incidence rate, cause various diseases and have various diseases, and the incidence rate of the anorectal diseases is on an increasing trend along with the improvement of the living standard of human beings. The anus and the intestine are important daily excretion channels for human beings, the diseased parts are special, the wound is easily polluted by excrement, the bacterial hyperplasia is difficult to heal, and the disease condition is easy to repeat. In recent years, surgery has become the main treatment means of anorectal diseases, aiming at improving the condition of the disease to the greatest extent and reducing the pain of patients, but due to the special diseased part, complications such as anal edema, wound infection, postoperative hematochezia and the like often occur after the surgery, so postoperative perioperative nursing is very important.
The invention relates to an antibacterial gel containing amphoteric alkaloid, wherein the amphoteric alkaloid is a nitrogen-containing organic compound simultaneously having amino and carboxyl, the molecule has a lactam structure, a primary amino group is obtained by partial hydrolysis, and the Schiff base compound and an active aldehyde group of oxidized sodium alginate have Schiff base cross-linking reaction, so that the formed Schiff base compound has good antifungal effect, can effectively inhibit superoxide radical, and has double antibacterial effect with the original quaternary ammonium group. The antibacterial gel has mild effect, can effectively inhibit bacterial growth and prevent wound infection, has tight intermolecular interaction of all components, has stable property and proper viscosity, can stay at the wound for a long time, has the effects of moisturizing, lubricating and cleaning the wound, maintains micro-ecological balance and effectively promotes healing.
Disclosure of Invention
The invention aims to provide a safe and efficient broad-spectrum antibacterial gel which is applied to protection of anorectal diseases and perioperative wound care, more effectively prevents wound infection and promotes wound healing.
In order to solve the above problems, the technical scheme of the invention is as follows: an antibacterial gel containing amphoteric alkaloids comprises the following raw materials in parts by weight: 1-3 parts of amphoteric alkaloid, 0.8-4.5 parts of oxidized sodium alginate, 0.5-2 parts of carbomer, 0.5-4 parts of triethanolamine, 7-13 parts of glycerol and 100 parts of purified water.
Preferably, the amphoteric alkaloid is a nitrogen-containing organic compound having a carboxyl group.
Preferably, the weight ratio of the carbomer to the triethanolamine is 1 (1-2).
Preferably, the weight ratio of the amphoteric alkaloid to the oxidized sodium alginate is 1 (0.8-1.5).
A preparation method of antibacterial gel containing amphoteric alkaloid comprises the following steps:
s1: preparing oxidized sodium alginate: weighing 1g of sodium alginate, dissolving the sodium alginate in purified water to obtain 1% sodium alginate solution, adding 1mL of 0.25mol/L sodium periodate solution, and oxidizing for 24h in a dark place; adding ethylene glycol, stirring for 15min to terminate the reaction, adding 0.3g of sodium chloride, stirring uniformly, adding 200mL of absolute ethanol to precipitate the product, performing suction filtration, dissolving with purified water, precipitating with ethanol, performing suction filtration, repeating for three times, and freeze-drying the product to obtain oxidized sodium alginate;
s2: dropwise adding acetic acid into 40-60 parts of purified water to adjust the pH value to be 5.0-5.5, adding 0.8-4.5 parts of oxidized sodium alginate and 1-3 parts of amphoteric alkaloid, switching on condensed water, and carrying out oil bath at a constant temperature of 90 ℃ for 30min to obtain an amphoteric alkaloid sodium alginate solution;
s3: spreading 0.5-2 parts of carbomer on 40-60 parts of purified water for multiple times, standing for 12 hours to fully swell the carbomer, and stirring uniformly to obtain a carbomer gel solution;
S4: dropwise adding 0.5-4 parts of triethanolamine into the solution obtained in the step S3, and stirring while adding to obtain a high-transparency gel solution;
s5: and (3) mixing the solution S4 with the solution S2, adding 7-13 parts of glycerol, and uniformly stirring in vacuum to obtain the glycerol-free oil.
The invention has the beneficial effects that:
(1) the gel prepared by the process has compact intermolecular interaction, and the gel matrix is transparent, uniform and fine; under the conditions of high temperature, low temperature and strong light, the appearance and the pH value of the gel are not obviously changed, and the product quality is stable and controllable.
(2) The antibacterial agent used for the antibacterial gel containing the amphoteric alkaloid has mild action, high safety and suitability for wounds, wherein the amphiphilic tail end of the amphoteric alkaloid is easy to combine with dirt and fragments, can effectively clean the wounds, has good antibacterial property and has the effect of preventing wound infection.
(3) The amphoteric alkaloid has a lactam structure, is partially hydrolyzed to obtain primary amine groups, and undergoes Schiff base crosslinking reaction with active aldehyde groups of oxidized sodium alginate to form a Schiff base compound, which has good antifungal effect, can effectively inhibit superoxide radicals, and forms double antibacterial action with the original quaternary amine groups, so that the antibacterial gel is more stable and has better use effect.
(4) The amino groups at two ends of the triethanolamine molecule can react with the carboxyl groups on the carbomer molecule, so that the carbomer intermolecular crosslinking is realized, the gel viscosity is greatly increased, when the ratio of the carboxyl groups to the amino groups is 1:1 to 1:2, the amino groups can be fully reacted, the viscosity of the gel cannot be influenced by continuously increasing the dosage of the triethanolamine, and the cytotoxicity is increased.
(5) An antibacterial gel containing amphoteric alkaloid has good biocompatibility and has effect of promoting wound healing;
(6) an antibacterial gel containing amphoteric alkaloid has effects of relieving inflammation, and can be applied to anorectal region to effectively inhibit anal swelling.
Detailed Description
The first embodiment is as follows:
an antibacterial gel containing amphoteric alkaloids comprises the following raw materials in parts by weight: 1 part of amphoteric alkaloid, 3 parts of oxidized sodium alginate, 1 part of carbomer, 1 part of triethanolamine, 10 parts of glycerol and 100 parts of purified water. The preparation method comprises the following steps:
s1: preparing oxidized sodium alginate: weighing 1g of sodium alginate, dissolving the sodium alginate in purified water to obtain 1% sodium alginate solution, adding 1mL of 0.25mol/L sodium periodate solution, and oxidizing for 24h in a dark place; adding ethylene glycol, stirring for 15min to terminate the reaction, adding 0.3g of sodium chloride, stirring uniformly, adding 200mL of absolute ethanol to precipitate the product, performing suction filtration, dissolving with purified water, precipitating with ethanol, performing suction filtration, repeating for three times, and freeze-drying the product to obtain oxidized sodium alginate;
S2: dropwise adding acetic acid into 50 parts of purified water to adjust the pH value to 5.0, adding 3 parts of oxidized sodium alginate and 1 part of amphoteric alkaloid, and boiling for 10min to obtain an amphoteric alkaloid sodium alginate solution;
s3: spreading 1 part of carbomer on 50 parts of purified water for multiple times, standing for 12h to fully swell the carbomer, and stirring uniformly to obtain carbomer gel solution;
s4: dropwise adding 1 part of triethanolamine into the solution in the S3, and stirring while adding to obtain a highly transparent gel solution;
s5: and (3) mixing the solution S4 with the solution S2, adding 10 parts of glycerol, and uniformly stirring in vacuum to obtain the glycerol-free oil.
Example two:
an antibacterial gel containing amphoteric alkaloids comprises the following raw materials in parts by weight: 2 parts of amphoteric alkaloid, 3 parts of oxidized sodium alginate, 1 part of carbomer, 1.5 parts of triethanolamine, 10 parts of glycerol and 100 parts of purified water. The preparation method comprises the following steps:
s1: preparing oxidized sodium alginate: weighing 1g of sodium alginate, dissolving the sodium alginate in purified water to obtain 1% sodium alginate solution, adding 1mL of 0.25mol/L sodium periodate solution, and oxidizing for 24h in a dark place; adding ethylene glycol, stirring for 15min to terminate the reaction, adding 0.3g of sodium chloride, stirring uniformly, adding 200mL of absolute ethanol to precipitate the product, performing suction filtration, dissolving with purified water, precipitating with ethanol, performing suction filtration, repeating for three times, and freeze-drying the product to obtain oxidized sodium alginate;
S2: dropwise adding acetic acid into 50 parts of purified water to adjust the pH value to 5.0, adding 3 parts of oxidized sodium alginate and 2 parts of amphoteric alkaloid, and boiling for 10min to obtain an amphoteric alkaloid sodium alginate solution;
s3: spreading 1 part of carbomer on 50 parts of purified water for multiple times, standing for 12h to fully swell the carbomer, and stirring uniformly to obtain carbomer gel solution;
s4: dropwise adding 1.5 parts of triethanolamine into the solution obtained in the step S3, and stirring while adding to obtain a high-transparency gel solution;
s5: and (3) mixing the solution S4 with the solution S2, adding 10 parts of glycerol, and uniformly stirring in vacuum to obtain the glycerol-free oil.
Example three:
an antibacterial gel containing amphoteric alkaloids comprises the following raw materials in parts by weight: 3 parts of amphoteric alkaloid, 3 parts of oxidized sodium alginate, 1 part of carbomer, 1 part of triethanolamine, 10 parts of glycerol and 100 parts of purified water. The preparation method comprises the following steps:
s1: preparing oxidized sodium alginate: weighing 1g of sodium alginate, dissolving the sodium alginate in purified water to obtain 1% sodium alginate solution, adding 1mL of 0.25mol/L sodium periodate solution, and oxidizing for 24h in a dark place; adding ethylene glycol, stirring for 15min to terminate the reaction, adding 0.3g of sodium chloride, stirring uniformly, adding 200mL of absolute ethanol to precipitate the product, performing suction filtration, dissolving with purified water, precipitating with ethanol, performing suction filtration, repeating for three times, and freeze-drying the product to obtain oxidized sodium alginate;
S2: dropwise adding acetic acid into 50 parts of purified water to adjust the pH value to 5.0, adding 3 parts of oxidized sodium alginate and 3 parts of amphoteric alkaloid, and boiling for 10min to obtain an amphoteric alkaloid sodium alginate solution;
s3: spreading 1 part of carbomer on 50 parts of purified water for multiple times, standing for 12h to fully swell the carbomer, and stirring uniformly to obtain carbomer gel solution;
s4: dropwise adding 1 part of triethanolamine into the solution in the S3, and stirring while adding to obtain a highly transparent gel solution;
s5: and (3) mixing the solution S4 with the solution S2, adding 10 parts of glycerol, and uniformly stirring in vacuum to obtain the glycerol-free oil.
Example four:
and (3) carrying out antibacterial performance detection on the antibacterial gel sample containing the amphoteric alkaloid prepared in the second example.
Streaking one of the preserved bacteria in nutrient agar, culturing at 37 ℃ for 24 hours, inoculating one of the preserved bacteria in 20mL of nutrient broth, performing shake culture at 37 ℃ for 18 hours at 130r/min, and diluting with PBS (0.03 mol/L) until the number of viable bacteria is 1 × 109 CFU/mL-5 × 109CFU/mL to obtain a bacterial suspension for later use. 95ml PBS buffer (0.03 mol/L) and 1g of sample were added to the flask and stirred uniformly, and a blank was set.
Adding 5mL of the bacterial suspension into a flask, uniformly shaking at constant temperature, immediately taking 1mL of the bacterial suspension, performing gradient dilution for four times by using PBS (phosphate buffer solution) and 10 times, coating 200 microliters of the fourth diluted bacterial suspension on a culture medium, performing constant-temperature back-off culture at 37 ℃ for 48 hours, and observing the antibacterial efficiency of the medical hydrogel after contact with the bacterial suspension 0.
The remaining solution in the flask was further shake-cultured at 37 ℃ at 130r/min for 4 and 8 hours, and then diluted with PBS in four 10-fold gradients. And (3) coating 200 microliters of the fourth diluted bacterial suspension on a culture medium, carrying out constant-temperature back-off culture at 37 ℃ for 48 hours, and observing the bacteriostasis efficiency of the medical hydrogel after the medical hydrogel is contacted with the bacterial suspension.
The results of the experiment are shown in table 1:
TABLE 1 results of antibacterial experiments
Contact time | Candida albicans | Escherichia coli | Staphylococcus aureus |
0h | 24.7% | 90.2% | 94.6% |
4h | 68.9% | 95.3% | 98.0% |
8h | 86.3% | 95.7% | 98.2% |
As can be seen from Table 1, the antibacterial gel containing the bioactive alkali has good antibacterial effects on Escherichia coli 8099, Staphylococcus aureus ATCC 6538 and Candida albicans ATCC 10231, wherein the antibacterial gel has relatively slow effect on Candida albicans ATCC 10231, but has rapid bactericidal effect on Escherichia coli 8099 and Staphylococcus aureus ATCC 6538,
example five:
because the influence of the antibacterial gel containing the amphoteric bioactive alkali on anorectal diseases is difficult to verify by directly modeling the animal anus, in order to verify the effectiveness of the antibacterial gel, a rat wound healing test and a rat anal swelling model test are selected for comprehensive analysis and verification.
(1) Rat wound healing test:
Animal strains were used: SD rat, clean grade;
animal sources: the production license number is SCXK (Guangdong) 2018-;
a breeding environment: the temperature of the SPF animal house is 20 +/-2 ℃, and the relative humidity is 20-30%.
The experimental steps are as follows: injecting anesthesia to a rat, shearing hairs on the back, wiping and disinfecting the skin by 75% alcohol, forming a 1 cm-by-1 cm circular wound at the position 4cm away from the median line of the back of the ear in the middle of the back of the rat by using a skin biopsy device with the diameter of 1cm, cutting the wound to reach fascia deeply, performing wound surface treatment, and observing the wound healing condition.
Treating a wound surface:
negative control group: the double-layer spun yarn is covered with a wound and bandaged after being soaked in normal saline.
Sample group: and (3) smearing the antibacterial gel of the second embodiment on the wound surface, and covering and wrapping the wound surface by using a double-layer spinning pack.
Positive control-1: the wound surface is smeared with a certain hemorrhoid ointment in the market, and is wrapped by a double-layer spinning.
Positive control-2: a certain carbomer gel product is coated on the surface of a wound and bound by a double-layer spinning pack.
The test results are shown in table 2 below:
table 2 rat wound test results
Experimental groups | Eighth balance average wound healing Rate |
Negative control group | 66.6% |
Sample set | 94.9% |
Positive control group-1 | 84.6% |
Positive control group-2 | 66.2% |
As can be seen from table 2, the antibacterial gel containing amphoteric alkaloid according to the present invention has a relatively significant effect of promoting wound healing compared to the positive control group.
Example four: rat anal swelling model test
Animal strains were used: SD rat, clean grade;
animal sources: the production license number is SCXK (Guangdong) 2018-;
a breeding environment: the temperature of the SPF animal house is 20 +/-2 ℃, and the relative humidity is 20-30%.
The experimental steps are as follows: weighing 0.8mL of distilled water, 4mL of pyridine 3.2mL of ether and 8mL of 6% croton oil, uniformly mixing to prepare an inflammation agent, soaking the inflammation agent in a cotton ball, inserting the cotton ball into a rat anus for 15 seconds, establishing a rat anus croton oil swelling model, observing the rat anus after 4 hours, and if the anus is red and swollen, and yellow secretion exists, determining that the model building is successful; after the molding is successful, the medicine is administrated, and the volume of each medicine is about 2.5ml (100 ml is used for each adult, adult: rat = 36: 1), 2 times per day; and measuring the whole perianal tissue swelling range by using a two-foot compass 4h and 48h after the molding, and calculating the swelling inhibition rate, wherein the test results are shown in the following table 3:
anal administration treatment
Negative control group: 2.5mL of physiological saline was injected into the anus of the rat.
Sample group: a2.5 mL sample of the antimicrobial gel was injected into the anus of the rat.
Positive control-1: 2.5mL of a hemorrhoid ointment from the market was injected into the anus of the rat.
Positive control-2: 2.5mL of a commercially available carbomer gel product was injected into the anus of the rat.
TABLE 3 rat anal swelling model test results
Experimental groups | Mean inhibition of swelling |
Negative control group | 62.4% |
Sample set | 85.5% |
Positive control group-1 | 71.5% |
Positive control group-2 | 71.1% |
As can be seen from Table 3, the amphoteric alkaloid-containing antibacterial gel of the present invention is more effective in inhibiting anal swelling than the positive control group.
Claims (5)
1. An antibacterial gel containing amphoteric alkaloids is characterized by comprising the following raw materials in parts by weight: 1-3 parts of amphoteric alkaloid, 0.8-4.5 parts of oxidized sodium alginate, 0.5-2 parts of carbomer, 0.5-4 parts of triethanolamine, 7-13 parts of glycerol and 100 parts of purified water.
2. The antibacterial gel containing amphoteric alkaloids according to claim 1, wherein the amphoteric alkaloids are nitrogen-containing organic compounds having carboxyl group.
3. The antibacterial gel containing amphoteric alkaloids according to claim 1, wherein the weight ratio of carbomer to triethanolamine is 1 (1-2).
4. The antibacterial gel containing amphoteric alkaloids according to claim 1, wherein the weight ratio of amphoteric alkaloids to sodium alginate oxide is 1 (0.8-1.5).
5. A process for the preparation of an antibacterial gel containing amphoteric alkaloids according to any one of claims 1 to 4, characterized in that it comprises the following steps:
s1: preparing oxidized sodium alginate: weighing 1g of sodium alginate, dissolving the sodium alginate in purified water to obtain a 1% sodium alginate solution, adding 1mL of 0.25mol/L sodium periodate solution, oxidizing for 24 hours in a dark place, adding ethylene glycol, stirring for 15min to terminate the reaction, adding 0.3g of sodium chloride, stirring uniformly, adding 200mL of absolute ethanol to precipitate a product, performing suction filtration, dissolving with purified water, precipitating with ethanol, performing suction filtration, repeating for three times, and freeze-drying the product to obtain oxidized sodium alginate;
s2: dropwise adding acetic acid into 40-60 parts of purified water to adjust the pH value to be 5.0-5.5, adding 0.8-4.5 parts of oxidized sodium alginate and 1-3 parts of amphoteric alkaloid, switching on condensed water, and carrying out oil bath at a constant temperature of 90 ℃ for 30min to obtain an amphoteric alkaloid sodium alginate solution;
s3: spreading 0.5-2 parts of carbomer on 40-60 parts of purified water for multiple times, standing for 12 hours to fully swell the carbomer, and stirring uniformly to obtain a carbomer gel solution;
s4: dropwise adding 0.5-4 parts of triethanolamine into the solution obtained in the step S3, and stirring while adding to obtain a high-transparency gel solution;
s5: and (3) mixing the solution S4 with the solution S2, adding 7-13 parts of glycerol, and uniformly stirring in vacuum to obtain the glycerol-free oil.
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