CN113768950B - Flushing fluid special for pets and preparation method thereof - Google Patents
Flushing fluid special for pets and preparation method thereof Download PDFInfo
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- CN113768950B CN113768950B CN202111069302.0A CN202111069302A CN113768950B CN 113768950 B CN113768950 B CN 113768950B CN 202111069302 A CN202111069302 A CN 202111069302A CN 113768950 B CN113768950 B CN 113768950B
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- pet
- chloride
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- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 claims abstract description 33
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Abstract
The invention discloses a special flushing fluid for pets and a preparation method thereof, wherein the flushing fluid comprises the following components in concentration: 120-180 mmol/L of sodium ions, 5-15 mmol/L of potassium ions, 0.5-3 mmol/L of magnesium ions, 1-5 mmol/L of calcium ions, 10-40 mmol/L of acetate ions and 1-10 mmol/L of citrate ions; 1-10 mmol/L of EDTA ions, 0.05-0.15 wt% of polyhexamethylene biguanide salt and 0.05-0.15 wt% of undecyleneamidopropyl betaine. The pH value of the flushing liquid special for the pet is 6.8-8.0, and the osmotic pressure value is 260-350 mOsm/kg. The components in the proportion have the functions of maintaining cell osmotic pressure, regulating and controlling the pH value balance of cell sap and inhibiting bacteria, promote the cell growth of pet wounds and are beneficial to the rapid healing of operation wounds.
Description
Technical Field
The invention relates to the technical field of medicines, and particularly relates to a flushing fluid special for pets and a preparation method thereof.
Background
Wound irrigation is a widely used therapeutic means, plays an important role in wound management, and can hydrate wounds, remove deep debris and assist medical staff in better wound examination. The irrigation fluid may remove cellular debris and surface microorganisms or residual topical drugs and dressings. Wound irrigation cleans the wound more effectively than cotton swab swabs or dips.
Normal wound healing is divided into 3 phases, the inflammatory, proliferative and epithelial phases, which are linked to each other. During normal healing, infectious microbes, exogenous debris and necrotic tissue are cleared during the inflammatory phase by the collective blood vessels and cells' response to the wound. However, insufficient inflammatory response can lead to ineffective clearance of pathogenic microorganisms by the body, resulting in impaired angiogenesis and granulation tissue growth, and infection. Pathogenic microorganisms can also affect collagen synthesis and matrix metalloproteinase function, and can also cause hypoxia and inhibit the normal function of neutrophils and macrophages.
Along with debridement, wound irrigation can effectively remove debris and pathogens from wounds, helping wound healing progress from the inflammatory phase to the proliferative phase. Wound irrigation, if performed properly, can promote healing of the wound from deep tissue to the skin surface. It also prevents false healing on purulent cysts or infected sinus surfaces. The goal of irrigation is to clean the wound while avoiding causing wound bed damage and reducing the risk of bacteria entering the deep tissue.
The selection of an appropriate irrigation fluid is a critical step in irrigation and fluids that may be used for topical irrigation include wound cleansers, antibiotics, antifungal drugs, antiseptic antiseptics, anesthetics, and the like. The ideal rinse solution should be isotonic, non-hemolytic, non-toxic, transparent, easily sterilizable, and inexpensive. Unfortunately, however, there is not such a perfect liquid. The current literature generally favors the use of saline as a flushing fluid. Many disinfectants and antibiotics are used as irrigation fluids, but the effectiveness is debated. The cytotoxicity of irrigation fluids should be considered, especially antibacterial disinfectants such as iodonium, chlorhexidine, and hydrogen peroxide, which may be toxic to tissues and detrimental to acute wound healing. The wound irrigation solution for clinical use comprises normal saline, distilled water, drinkable tap water, disinfectant (such as povidone iodine, chlorhexidine, hydrogen peroxide and sodium hypochlorite), surfactant (such as soap water, benzalkonium chloride, etc.), antibacterial drug (metronidazole, ornidazole, etc.), etc. At present, no wound irrigation solution specially for pets (cats and dogs) exists, and the wound irrigation solution for pets in clinical use is usually directly replaced by the wound irrigation solution for pets.
Most mammalian cells tolerate osmoses of 260-350mOsm, and animal cell osmoses are also distinguished from human plasma osmoses. For example: the human plasma osmotic pressure of 290mQsm/kg can be regarded as the ideal osmotic pressure for culturing human cells; the osmotic pressure of the rat cells was around 320 mOsm/kg. In addition, fibroblasts critical for wound healing in animals are more suitable for mildly alkaline environments with pH 7.4-7.7.
Considering that the wound irrigation solution for pets is wide in application and strong in pertinence of the using groups, how to prepare the special irrigation solution for pets, which is isotonic with animal cells, non-hemolytic, non-toxic, transparent, easy to disinfect, economical and environment-friendly, suitable for wound treatment and promotion of wound cell growth, becomes a technical requirement in the field.
Disclosure of Invention
The invention aims to provide a special washing liquid for pets, which is suitable for wound treatment and promotes wound cell growth.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect of the present invention, there is provided a pet-specific washing liquid comprising the following components in concentration: 120-180 mmol/L of sodium ions, 5-15 mmol/L of potassium ions, 0.5-3 mmol/L of magnesium ions, 1-5 mmol/L of calcium ions, 10-40 mmol/L of acetate ions and 1-10 mmol/L of citrate ions.
Further, the special pet flushing liquid comprises the following components in concentration: 140-160 mmol/L of sodium ions, 8-12 mmol/L of potassium ions, 1.0-2.0 mmol/L of magnesium ions, 2-4 mmol/L of calcium ions, 15-35 mmol/L of acetate ions and 4-7 mmol/L of citrate ions.
Further, the flushing liquid special for the pet comprises the following components in concentration: 154.2mmol/L sodium ion, 10mmol/L potassium ion, 1.48mmol/L magnesium ion, 3.2mmol/L calcium ion, 28.8mmol/L acetate, 5.6mmol/L citrate ion.
Further, the flushing liquid special for the pet also comprises the following components in concentration: EDTA 1-10 mmol/L, polyhexamethylene biguanide salt 0.05-0.15 wt% and undecylenic amido propyl betaine 0.05-0.15 wt%.
Further, the special pet flushing fluid also comprises the following components in concentration: EDTA 2-8 mmol/L, polyhexamethylene biguanide salt 0.08-0.12 wt% and undecylenic amide propyl betaine 0.08-0.12 wt%.
Further, the flushing liquid special for the pet also comprises the following components in concentration: EDTA5mmol/L, polyhexamethylene biguanide salt 0.1wt% and undecylenamidopropyl betaine 0.1wt%.
The pH value of the flushing liquid special for the pet is 6.8-8.0, and the osmotic pressure value is 260-350 mOsm/kg; further, the pH value of the flushing liquid special for the pet is 7.0-7.7, and the osmotic pressure value is 290-330 mOsm/kg.
In a second aspect of the present invention, there is provided a method for preparing a pet-specific rinse solution, the method comprising:
dissolving the components (sodium chloride, potassium chloride, calcium chloride, magnesium chloride, sodium acetate, sodium citrate, EDTA disodium, polyhexamethylene biguanide salt and undeceneamidopropyl betaine) of the flushing fluid special for the pet in purified water, adjusting the pH value, and sterilizing to obtain the flushing fluid special for the pet.
Further, the sterilizing comprises: moist heat sterilization at 121 deg.C for 30min.
One or more technical solutions in the embodiments of the present invention at least have the following technical effects or advantages:
1. experiments show that the ratio (120-180 mmol/L of sodium ions, 5-15 mmol/L of potassium ions, 0.5-3 mmol/L of magnesium ions, 1-5 mmol/L of calcium ions, 10-40 mmol/L of acetate ions and 1-10 mmol/L of citrate ions) of the flushing liquid special for pets can reach the pH value in the range of 6.8-8.0 and the osmotic pressure in the range of 260-350 mOsm/kg; the components in the proportion are closest to extracellular fluid components, have the minimum interference on the internal environment of body fluid of an organism, have the functions of maintaining the osmotic pressure of cells and regulating and controlling the pH value balance of cell sap, are suitable for wound treatment and promotion of the growth of wound cells, and are beneficial to the rapid healing of surgical wounds.
2. The special flushing fluid for pets, provided by the invention, is added with 1-10 mmol/L of EDTA, 0.05-0.15 wt% of polyhexamethylene biguanide salt and 0.05-0.15 wt% of undecylene amidopropyl betaine, and is in synergistic cooperation, so that the flushing fluid has an obvious synergistic and additive effect, and can specifically kill pathogenic microorganisms such as staphylococcus aureus, escherichia coli, bacillus subtilis, pseudomonas aeruginosa and candida albicans. Meanwhile, the wound healing agent has no or small killing and inhibiting effects on normal cells, and can obviously prevent wound infection, thereby further improving the wound healing rate.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on the drawings without creative efforts.
FIG. 1 is a diagram showing the bacteriostatic effects of a pet-specific rinse solution and physiological saline provided in examples 2, 4 to 7 and 14 of the present invention;
FIG. 2 is a diagram illustrating the bacteriostatic effects of the pet-specific rinse solution and physiological saline according to examples 8 to 13 of the present invention;
fig. 3 is a diagram illustrating the bacteriostatic effects of the pet-specific rinse solution, the physiological saline and the ornidazole provided in examples 10 and 12 to 13 of the present invention;
FIG. 4 is a photograph of the bacteriostatic circle of Pseudomonas aeruginosa, staphylococcus aureus, escherichia coli, bacillus subtilis and Candida albicans for the prescription in an embodiment of the present invention;
FIG. 5 is a diagram illustrating the safety effect of the washing liquid for pets on immortalized epidermal cells;
FIG. 6 is a diagram showing the effect of the rinse solution for pets on the growth of immortalized epidermal cells;
FIG. 7 is a diagram showing the therapeutic effect of the rinse solution for pets according to the present invention.
Detailed Description
The present invention will be described in detail below with reference to specific embodiments and examples, and the advantages and various effects of the present invention will be more clearly apparent therefrom. It will be understood by those skilled in the art that these specific embodiments and examples are for the purpose of illustrating the invention and are not to be construed as limiting the invention.
Throughout the specification, unless otherwise specifically noted, terms used herein should be understood as having meanings as commonly used in the art. Accordingly, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. If there is a conflict, the present specification will control.
Unless otherwise specifically stated, various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or can be obtained by existing methods.
In order to solve the technical problems, the general idea of the embodiment of the application is as follows:
the application firstly analyzes the reason that the special washing liquid for pets in the prior art has the problems that the wound treatment suitability is small and the growth of wound cells cannot be promoted or cannot be promoted to a small extent:
in the market, normal saline is mostly adopted for wound irrigation solution for human and animals. The normal saline is a solution only containing sodium ions and chloride ions, has a large difference from normal plasma components, and is easy to cause acid-base balance, electrolyte disorder and kidney injury of a human body due to overhigh content of chloride ions. Although the animal body has stronger compensation and metabolic conversion capability, when the body organs, especially the kidney, are damaged, the body compensation capacity is reduced (young, disabled state, etc.), or the body is rapidly infused in a large amount in a short time, so that the body organs and other drugs or metabolites are discharged from the kidney in a large amount competitive manner, the kidney of the animal is difficult to effectively discharge the excessive chloride ions in the plasma in time. At present, the special operation wound washing liquid for pets is also lacked in the market.
Therefore, how to develop a special pet flushing fluid can reach the osmotic pressure of 260-350 mOsm/kg; the components in the proportion are closest to extracellular fluid components, have minimum interference on the internal environment of body fluid of an organism, have the functions of maintaining the osmotic pressure of cells and regulating and controlling the pH value balance of cell sap, and are suitable for wound treatment and promotion of the growth of wound cells.
According to an exemplary embodiment of the present invention, there is provided a pet-specific washing solution, which includes the following components in concentration: 120-180 mmol/L of sodium ions, 5-15 mmol/L of potassium ions, 0.5-3 mmol/L of magnesium ions, 1-5 mmol/L of calcium ions, 10-40 mmol/L of acetate ions and 1-10 mmol/L of citrate ions;
the pH value of the flushing liquid special for the pet is 6.8-8.0, and the osmotic pressure value is 280-330 mOsm/kg.
In the invention, experiments show that the osmotic pressure of the mixture ratio (120-180 mmol/L of sodium ions, 5-15 mmol/L of potassium ions, 0.5-3 mmol/L of magnesium ions, 1-5 mmol/L of calcium ions, 10-40 mmol/L of acetate ions and 1-10 mmol/L of citrate ions) can reach the range of 260-330 mOsm/kg; the components in the proportion are closest to extracellular fluid components, have minimum interference on the internal environment of body fluid of an organism, have the functions of maintaining the osmotic pressure of cells and regulating and controlling the pH value balance of cell sap, and are suitable for wound treatment and promotion of the growth of wound cells; specifically, the method comprises the following steps:
120-180 mmol/L of sodium ions: sodium is the major extracellular ion in plasma and aqueous humor and is essential for maintaining cell tone.
10-30 mmol/L of potassium ions: potassium is the major ion in the cell.
K + influx and Na + efflux are regulated by Na + K + -ATPase, actively spanning the membrane to maintain intracellular potentials. Too high or too low sodium ion concentration and potassium ion concentration are not favorable for maintaining osmotic pressure in the range of 260-330 mOsm/kg, so that water and electrolyte in intracellular and extracellular fluids are disturbed, and cells are broken and die.
Magnesium ion 0.5-3 mmol/L: magnesium is one of the major intracellular cations in mammals and is a cofactor for certain a + D enzymes and many cellular biochemical reactions. Too high or too low concentration of magnesium ions is not favorable for regulating energy consumption of cells and influencing growth of cells.
1-5 mmol/L of calcium ions: the existence of calcium ions has certain promotion effect on maintaining the connection function between endothelial cells. Too low a concentration of calcium ions is detrimental to cell motility.
1-10 mmol/L of citrate radical ion: the citrate ion and the calcium ion in blood form a dissociable soluble complex calcium citrate, which is easily soluble in water but not dissociable, so that the calcium ion in blood is reduced, the blood coagulation process is inhibited, and the blood coagulation is prevented. Excessive or insufficient concentrations of citric acid and/or sodium citrate are not favorable for keeping the cation concentration balance inside and outside the cells.
10-40 mmol/L of acetate ions: the acetate ion can be used for slightly resisting systemic acidosis. And too high or too low acetate concentration is not favorable for accelerating cell metabolism.
Preferably, the pet-specific washing liquid comprises the following components in concentration: 140-160 mmol/L of sodium ions, 8-12 mmol/L of potassium ions, 1.0-2.0 mmol/L of magnesium ions, 2-4 mmol/L of calcium ions, 15-35 mmol/L of acetate ions and 4-7 mmol/L of citrate ions; the pH value of the flushing liquid special for the pet is 7.0-7.7, and the osmotic pressure value is 280-330 mOsm/kg.
More preferably, the pet-specific washing solution comprises the following components in concentration: 154.2mmol/L sodium ion, 10mmol/L potassium ion, 1.48mmol/L magnesium ion, 3.2mmol/L calcium ion, 28.8mmol/L acetate, 5.6mmol/L citrate ion; the pH value of the flushing liquid special for the pet is 7.0-7.4, and the osmotic pressure value is 280-330 mOsm/kg.
In addition to maintaining wound cell growth, preventing microbial growth and infection at the wound is another need for irrigation fluids. The conventional chemical antibiotics can kill pathogenic microorganisms and can kill and inhibit normal cells. Therefore, how to develop a bacteriostatic component capable of specifically killing pathogenic microorganisms and simultaneously having no or less killing and inhibiting effects on normal cells becomes a technical problem.
Therefore, according to another exemplary embodiment of the present invention, there is provided a pet-specific washing solution comprising, in terms of concentration, the following components: 140-160 mmol/L of sodium ions, 8-12 mmol/L of potassium ions, 1.0-2.0 mmol/L of magnesium ions, 2-4 mmol/L of calcium ions, 15-35 mmol/L of acetate ions and 4-7 mmol/L of citrate ions;
the pH value of the flushing liquid special for the pet is 6.8-8.0, and the osmotic pressure value is 260-350 mOsm/kg;
further comprising: EDTA 1-10 mmol/L, polyhexamethylene biguanide salt 0.05-0.15 wt% and undecylenic amido propyl betaine 0.05-0.15 wt%.
The embodiment of the invention discovers through experiments that: adding 1-10 mmol/L of EDTA, 0.05-0.15 wt% of polyhexamethylene biguanide salt and 0.05-0.15 wt% of undecylenamidopropyl betaine, and synergistically matching to kill pathogenic microorganisms specifically and have no or less killing and inhibiting effects on normal cells.
EDTA 1-10 mmol/L: EDTA itself is also an antibacterial and antibiofilm agent. For the treatment of biofilm-related diseases in dentistry, medical devices and veterinary medicine. In veterinary medicine, EDTA solutions can be used to treat biofilm-related infections in the ears of dogs. Furthermore, EDTA can also be used as an osmotic and sensitizing agent in combination with various other active ingredients. Experiments show that EDTA can not specifically kill pathogenic microorganisms when used alone, and does not have or has small killing and inhibiting effects on normal cells when being cooperated with other two bacteriostatic components in the range to specifically kill the pathogenic microorganisms. Too large or too small a concentration makes it difficult to achieve other effects described above.
0.05-0.15 wt% of polyhexamethylene biguanide salt: polyhexamethylene biguanidine (PHMB Polyhexamethylene biguanidine cas: 57029-18-2) has a broad spectrum of bactericidal activity, including gram positive and gram negative bacteria, as well as some fungi and resistant microorganisms, such as MRSA. In vitro studies have also demonstrated that PHMB is also effective against bacteria in biofilms. And has excellent biocompatibility. In addition, due to the diversity of the mechanism of action of PHMB, the potential for resistance is minimal. Experiments show that PHMB cannot specifically kill pathogenic microorganisms when being used alone, and does not have or have small killing and inhibiting effects on normal cells when being matched with other two bacteriostatic components in the range to specifically kill the pathogenic microorganisms. Too large or too small a concentration makes it difficult to achieve other effects.
0.05 to 0.15wt% of undecylenamidopropylbetaine: in wound management, irrigation of wounds is to reduce bacterial biofilm and prevent biofilm remodeling, such as polyhexamethylene biguanide and surfactants (undecenamidopropylbetaine) can significantly enhance penetration of difficult to remove wound coverings, enhancing the ability to remove wound tissue debris, bacteria and biofilm; can reduce surface tension and promote the removal of bacteria and debris during lavage, while the conventional 0.9% sodium chloride solution can only slide across the surface of the biofilm and can not destroy and remove the biofilm. Experiments show that the undeceneamidopropyl betaine can not specifically kill pathogenic microorganisms when being used alone, and the undeceneamidopropyl betaine is synergistically matched with other two antibacterial components within the range to specifically kill the pathogenic microorganisms, and has no or less killing and inhibiting effects on normal cells. Too large or too small a concentration makes it difficult to achieve other effects.
Preferably, the special pet flushing liquid further comprises the following components in concentration: EDTA 2-8 mmol/L, polyhexamethylene biguanide salt 0.08-0.12 wt% and undecylenic amide propyl betaine 0.08-0.12 wt%.
More preferably, the special pet washing liquid further comprises the following components in concentration: EDTA5mmol/L, polyhexamethylene biguanide salt 0.1wt% and undecylenamidopropylbetaine 0.1wt%.
According to another typical embodiment of the invention, the preparation method of the special pet washing liquid is provided, and the special pet washing liquid is prepared by dissolving the components (sodium chloride, potassium chloride, calcium chloride, magnesium chloride, sodium acetate, sodium citrate, disodium EDTA, polyhexamethylene biguanide salt and undecenylamidopropylbetaine) of the special pet washing liquid in purified water, adjusting the pH value and sterilizing the solution.
As a specific embodiment, the sterilizing comprises: sterilizing at 121 deg.C for 30min.
A pet rinse solution of the present application will be described in detail with reference to examples, comparative examples, and experimental data. The concentrations of the bacteriostatic agents of the embodiment and the comparative example are within the national standard range.
Example 1
This example provides a pet rinse solution, which is an aqueous solution with a ph of 6.8-8.0, and contains the following components: 154.2mmol/L sodium ion, 10mmol/L potassium ion, 1.476mmol/L magnesium ion, 3.2mmol/L calcium ion, 28.8mmol/L acetate ion, 5.6mmol/L citrate ion.
The preparation method of the rinse solution of the present embodiment includes the following steps:
weighing 6.40g of sodium chloride, 0.74g of potassium chloride, 0.36g of anhydrous calcium chloride, 0.14g of anhydrous magnesium chloride, 2.36g of anhydrous sodium acetate and 1.70g of sodium citrate dihydrate; preparing 1000ml solution with purified water, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30min.
Example 2
This example provides a pet rinse solution, which is an aqueous solution with a ph of 6.8-8.0, and contains the following components: 180mmol/L of sodium ions, 15mmol/L of potassium ions, 3mmol/L of magnesium ions, 5mmol/L of calcium ions, 50mmol/L of acetate ions and 10mmol/L of citrate ions.
The preparation method of the rinse solution of the present embodiment includes the following steps:
weighing 5.85g of sodium chloride, 1.12g of potassium chloride, 0.56g of anhydrous calcium chloride, 0.28g of anhydrous magnesium chloride, 4.10g of anhydrous sodium acetate and 3.03g of sodium citrate dihydrate; preparing 1000ml solution with purified water, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30min.
Example 3
This example provides a pet rinse solution, which is an aqueous solution with a ph of 6.8-8.0, and contains the following components: 120mmol/L of sodium ions, 5mmol/L of potassium ions, 0.5mmol/L of magnesium ions, 1mmol/L of calcium ions, 10mmol/L of acetate ions and 1mmol/L of citrate ions.
The preparation method of the rinse solution of the present embodiment includes the following steps:
weighing 6.26g of sodium chloride, 0.37g of potassium chloride, 0.112g of anhydrous calcium chloride, 0.05g of anhydrous magnesium chloride, 0.82g of anhydrous sodium acetate and 0.30g of sodium citrate dihydrate; preparing 1000ml solution with purified water, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30min.
Comparative example 1
In this comparative example, the rinsing solution was physiological saline (0.9% NaCl solution).
Comparative example 2
In this comparative example, the following components were contained: 300mmol/L of sodium ions, 45mmol/L of potassium ions, 6mmol/L of magnesium ions, 7.5mmol/L of calcium ions, 75mmol/L of acetate ions and 15mmol/L of citrate ions.
The preparation method of the rinse solution of the embodiment includes the following steps:
weighing 11.82g of sodium chloride, 3.33g of potassium chloride, 0.84g of anhydrous calcium chloride, 060g of anhydrous magnesium chloride, 6.15g of anhydrous sodium acetate and 4.5g of sodium citrate dihydrate; preparing 1000ml solution with purified water, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30min.
Example 4
This example provides a rinse solution, which is an aqueous solution having a pH of 6.8 to 8.0, and contains the following components: 154.2mmol/L sodium ion, 10mmol/L potassium ion, 1.476mmol/L magnesium ion, 3.2mmol/L calcium ion, 28.8mmol/L acetate ion, 5.6mmol/L citrate ion, 0.1% polyhexamethylene biguanide salt.
The preparation method of the rinse solution of the embodiment includes the following steps:
weighing 6.4g of sodium chloride, 0.74g of potassium chloride, 0.35g of anhydrous calcium chloride, 0.14g of anhydrous magnesium chloride, 2.36g of anhydrous sodium acetate, 1.70g of sodium citrate dihydrate and 1.0g of polyhexamethylene biguanide hydrochloride; preparing 1000ml solution with purified water, adjusting pH to 7.0-7.7, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30min.
Example 5
This example provides a rinse solution, which is an aqueous solution with a pH of 6.8 to 8.0, and contains the following components: 154.2mmol/L sodium ion, 10mmol/L potassium ion, 1.476mmol/L magnesium ion, 3.2mmol/L calcium ion, 28.8mmol/L acetate ion, 5.6mmol/L citrate ion and 1mmol/L EDTA ion.
The preparation method of the rinse solution of the embodiment includes the following steps:
weighing 6.19g of sodium chloride, 0.74g of potassium chloride, 0.36g of anhydrous calcium chloride, 0.14g of anhydrous magnesium chloride, 2.36g of anhydrous sodium acetate, 1.70g of sodium citrate dihydrate and 0.372g of EDTA disodium dihydrate; preparing 1000ml solution with purified water, adjusting pH to 7.0-7.7, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30min.
Example 6
This example provides a rinse solution, which is an aqueous solution having a pH of 6.8 to 8.0, comprising the following components: 154.2mmol/L sodium ion, 10mmol/L potassium ion, 1.476mmol/L magnesium ion, 3.2mmol/L calcium ion, 28.8mmol/L acetate ion, 5.6mmol/L citrate ion and 0.1% undecylenamidopropyl betaine.
The preparation method of the rinse solution of the present embodiment includes the following steps:
weighing 6.4g of sodium chloride, 0.74g of potassium chloride, 0.35g of anhydrous calcium chloride, 0.14g of anhydrous magnesium chloride, 2.36g of anhydrous sodium acetate, 1.70g of sodium citrate dihydrate and 1.0g of undecylenamidopropyl betaine; adjusting pH to about 7.0, preparing into 1000ml solution with purified water, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30min.
Example 7
This example provides a rinse solution, which is an aqueous solution having a pH of 6.8 to 8.0, comprising the following components: 154.2mmol/L of sodium ions, 10mmol/L of potassium ions, 1.476mmol/L of magnesium ions, 3.2mmol/L of calcium ions, 28.8mmol/L of acetate ions, 5.6mmol/L of citrate ions, 0.1 percent of polyhexamethylene biguanide salt and 1mmol/L of EDTA.
The preparation method of the rinse solution of the embodiment includes the following steps:
weighing 6.19g of sodium chloride, 0.74g of potassium chloride, 0.36g of anhydrous calcium chloride, 0.14g of anhydrous magnesium chloride, 2.36g of anhydrous sodium acetate, 1.70g of sodium citrate dihydrate, 0.372g of EDTA disodium dihydrate and 1.0g of polyhexamethylene biguanide hydrochloride; adjusting pH to about 7.0, preparing into 1000ml solution with purified water, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30min.
Example 8
This example provides a rinse solution, which is an aqueous solution having a pH of 6.8 to 8.0, comprising the following components: 154.2mmol/L sodium ion, 10mmol/L potassium ion, 1.476mmol/L magnesium ion, 3.2mmol/L calcium ion, 28.8mmol/L acetate ion, 5.6mmol/L citrate ion, 0.1% polyhexamethylene biguanide salt, 1mmol/L EDTA, and 0.1% undecenylamidopropyl betaine.
The preparation method of the rinse solution of the present embodiment includes the following steps:
weighing 6.19g of sodium chloride, 0.74g of potassium chloride, 0.36g of anhydrous calcium chloride, 0.14g of anhydrous magnesium chloride, 2.36g of anhydrous sodium acetate, 1.70g of sodium citrate dihydrate, 0.372g of EDTA disodium dihydrate, 1.0g of polyhexamethylene biguanide hydrochloride and 1.0g of undecylenamidopropyl betaine; adjusting pH to about 7.0, preparing into 1000ml solution with purified water, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30min.
Example 9
This example provides a rinse solution, which is an aqueous solution having a pH of 6.8 to 8.0, comprising the following components: 154.2mmol/L sodium ion, 10mmol/L potassium ion, 1.476mmol/L magnesium ion, 3.2mmol/L calcium ion, 28.8mmol/L acetate ion, 5.6mmol/L citrate ion, 0.1% polyhexamethylene biguanide salt, 2mmol/L EDTA, and 0.1% undecenylamidopropyl betaine.
The preparation method of the rinse solution of the present embodiment includes the following steps:
weighing 6.08g of sodium chloride, 0.74g of potassium chloride, 0.36g of anhydrous calcium chloride, 0.14g of anhydrous magnesium chloride, 2.36g of anhydrous sodium acetate, 1.70g of sodium citrate dihydrate, 0.744g of EDTA disodium dihydrate, 1.0g of polyhexamethylene biguanide hydrochloride and 1.0g of undecylenamidopropyl betaine; adjusting pH to about 7.0, preparing into 1000ml solution with purified water, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30min.
Example 10
This example provides a rinse solution, which is an aqueous solution having a pH of 6.8 to 8.0, comprising the following components: 154.2mmol/L sodium ion, 10mmol/L potassium ion, 1.476mmol/L magnesium ion, 3.2mmol/L calcium ion, 28.8mmol/L acetate ion, 5.6mmol/L citrate ion, 0.1% polyhexamethylene biguanide salt, 5mmol/L EDTA, and 0.1% undecenylamidopropyl betaine.
The preparation method of the rinse solution of the embodiment includes the following steps:
weighing 5.73g of sodium chloride, 0.74g of potassium chloride, 0.36g of anhydrous calcium chloride, 0.14g of anhydrous magnesium chloride, 2.36g of anhydrous sodium acetate, 1.70g of sodium citrate dihydrate, 1.86g of EDTA disodium dihydrate, 1.0g of polyhexamethylene biguanide hydrochloride and 1.0g of undecylenamidopropyl betaine; adjusting pH to about 7.0, preparing into 1000ml solution with purified water, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30min.
Example 11
This example provides a washing solution, which is an aqueous solution with a pH of 6.8-8.0, comprising the following components: 154.2mmol/L of sodium ions, 10mmol/L of potassium ions, 1.476mmol/L of magnesium ions, 3.2mmol/L of calcium ions, 28.8mmol/L of acetate ions, 5.6mmol/L of citrate ions, 0.1% of polyhexamethylene biguanide salt, 10mmol/L of EDTA and 0.1% of undecylenamidopropyl betaine.
The preparation method of the rinse solution of the present embodiment includes the following steps:
weighing 5.15g of sodium chloride, 0.74g of potassium chloride, 0.36g of anhydrous calcium chloride, 0.14g of anhydrous magnesium chloride, 2.36g of anhydrous sodium acetate, 1.70g of sodium citrate dihydrate, 3.72g of EDTA disodium dihydrate, 1.0g of polyhexamethylene biguanide hydrochloride and 1.0g of undecylenamidopropyl betaine; adjusting pH to about 7.0, preparing into 1000ml solution with purified water, filtering with 0.45 μm filter membrane, and wet-heat sterilizing at 121 deg.C for 30min.
Example 12
This example provides a washing solution, which is an aqueous solution with a pH of 6.8-8.0, comprising the following components: 154.2mmol/L of sodium ions, 10mmol/L of potassium ions, 1.476mmol/L of magnesium ions, 3.2mmol/L of calcium ions, 28.8mmol/L of acetate ions, 5.6mmol/L of citrate ions, 0.05% of polyhexamethylene biguanide salt, 0.5mmol/L of EDTA and 0.05% of undecylenamidopropyl betaine.
The preparation method of the rinse solution of the present embodiment includes the following steps:
weighing 6.26g of sodium chloride, 0.74g of potassium chloride, 0.36g of anhydrous calcium chloride, 0.14g of anhydrous magnesium chloride, 2.36g of anhydrous sodium acetate, 1.70g of sodium citrate dihydrate, 0.186g of EDTA disodium dihydrate, 1.0g of polyhexamethylene biguanide hydrochloride and 1.0g of undecylenamidopropyl betaine; adjusting pH to about 7.0, preparing into 1000ml solution with purified water, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30min.
Example 13
This example provides a washing solution, which is an aqueous solution with a pH of 6.8-8.0, comprising the following components: 154.2mmol/L of sodium ions, 10mmol/L of potassium ions, 1.476mmol/L of magnesium ions, 3.2mmol/L of calcium ions, 28.8mmol/L of acetate ions, 5.6mmol/L of citrate ions, 0.15% of polyhexamethylene biguanide salt, 5mmol/L of EDTA and 0.15% of undecylenamidopropyl betaine.
The preparation method of the rinse solution of the present embodiment includes the following steps:
weighing 5.73g of sodium chloride, 0.74g of potassium chloride, 0.36g of anhydrous calcium chloride, 0.14g of anhydrous magnesium chloride, 2.36g of anhydrous sodium acetate, 1.70g of sodium citrate dihydrate, 1.86g of EDTA disodium dihydrate, 1.0g of polyhexamethylene biguanide hydrochloride and 1.0g of undecylenamidopropyl betaine; adjusting pH to about 7.0, preparing into 1000ml solution with purified water, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30min.
Example 14
This example provides a rinse solution, which is an aqueous solution having a pH of 6.8 to 8.0, comprising the following components: 154.2mmol/L sodium ion, 10mmol/L potassium ion, 1.476mmol/L magnesium ion, 3.2mmol/L calcium ion, 28.8mmol/L acetate ion, 5.6mmol/L citrate ion, 0.1% polyhexamethylene biguanide salt and 5mmol/L EDTA.
The preparation method of the rinse solution of the embodiment includes the following steps:
weighing 5.73g of sodium chloride, 0.74g of potassium chloride, 0.36g of anhydrous calcium chloride, 0.14g of anhydrous magnesium chloride, 2.36g of anhydrous sodium acetate, 1.70g of sodium citrate dihydrate, 1.86g of EDTA disodium dihydrate and 1.0g of polyhexamethylene biguanide hydrochloride; adjusting pH to about 7.0, preparing into 1000ml solution with purified water, filtering with 0.45 μm filter membrane, and wet-heat sterilizing at 121 deg.C for 30min.
Experimental example 1
1. A summary of the formulations for each example and comparative example is shown in table 1;
table 1: examples and comparative summaries
From the data in table 1, it can be seen that:
in examples 1 to 3, the sodium ion concentration was 120 to 180mmol/L, the potassium ion concentration was 5 to 15mmol/L, the magnesium ion concentration was 0.5 to 3mmol/L, the calcium ion concentration was 1 to 5mmol/L, the acetate ion concentration was 10 to 40mmol/L, the citrate ion concentration was 1 to 10mmol/L, the pH range was 6.8 to 8.0, and the osmotic pressure was 260 to 350mOsm/kg, which are similar to the physiological saline of comparative example 1.
In comparative example 2, outside the above concentration range, the osmotic pressure also exceeded the expected range.
2. Bacteriostatic test
The bacteriostasis experiment adopts a filter paper method, adopts staphylococcus aureus, escherichia coli, bacillus subtilis, pseudomonas aeruginosa and candida albicans to investigate the bacteriostasis of the prescription, and has the dosage of 10 mu L.
For the screened prescription, two-fold dilutions of 4 gradients were performed, setting two parallel sets for a total of 18 dishes. The appearance of the bacteriostatic circle is shown in the attached figure 1: statistics of the data are shown in table 2;
table 2: bacillus subtilis bacteriostasis circle diameter (mm)
Table 3: diameter of escherichia coli inhibition zone (mm)
Table 4: diameter of antibacterial ring of staphylococcus aureus (mm)
Table 5: diameter of pseudomonas aeruginosa inhibition zone (mm)
Table 6: candida albicans antibacterial circle diameter (mm)
The data histograms of tables 2-6 are shown in FIGS. 1-3.
As can be seen from FIG. 1: both example 2 and saline were not bacteriostatic; both example 5 and example 6 were not bacteriostatic, indicating that neither EDTA or undecenylamidopropyl betaine alone was bacteriostatic;
example 4, example 7 and example 14 all were bacteriostatic; the polyhexamethylene biguanide salt is an essential bacteriostatic component; the bacteriostatic ratios of example 7 and example 14 are obviously enhanced compared with example 4, which shows that EDTA and polyhexamethylene biguanide salt are synergistic, and EDTA has obvious bacteriostatic enhancement effect.
As can be seen from fig. 2: examples 8-13 are all bacteriostatic; the polyhexamethylene biguanide salt, EDTA and undecylenic amide propyl betaine are added into the bacteriostatic component, the bacteriostatic effect is greatly superior to that of the mixture obtained in examples 4, 7 and 14, and the synergistic effect of the undecylenic amide propyl betaine, the EDTA and the polyhexamethylene biguanide salt is demonstrated, so that the bacteriostatic effect is enhanced. And shows that the antibacterial activity is increased along with the increase of the amount of EDTA.
As can be seen in FIG. 3: examples 10 to 13 all had bacteriostatic activity and were superior to the ornidazole control group (ornidazole sodium chloride solution, prepared by weighing 0.85g of sodium chloride and 0.5g of ornidazole, preparing 100ml of solution with purified water, filtering with 0.45 μm filter membrane, and sterilizing at 121 deg.C for 30 min). The antibacterial effect is influenced by the change of the amount of the polyhexamethylene biguanide salt, and the change is not large at the concentration of more than 0.1 percent.
Performing one-way anova on the drug group and the control group by using GraphPad Prism, and obtaining that p is less than 0.05, which indicates that the drug and the control group have significant difference;
the results of the bacteriostasis test show that: adding 1-10 mmol/L of EDTA, 0.05-0.15 wt% of polyhexamethylene biguanide salt and 0.05-0.15 wt% of undecenylamidopropylbetaine into the mixture, and performing synergistic cooperation, so that the composition has an obvious synergistic and additive effect, and can specifically kill pathogenic microorganisms such as staphylococcus aureus, escherichia coli, bacillus subtilis, pseudomonas aeruginosa and candida albicans.
3. Cell survival and growth experiments (MTT experiments)
The specific operation is as follows: a culture solution containing 10% fetal calf serum is prepared into a single HaCat (immortalized epidermal cell) cell suspension, and 1000-10000 cells per hole are inoculated into a 96-hole plate, wherein the hole volume is 200 mu l. After 3-5 days of incubation, the media (example 13) was dosed with 100. Mu.l/well and 20. Mu.l/well MTT thiazole blue solution (5 mg/ml in PBS). Incubation was continued for 4 hours, the culture was terminated, and the culture supernatant in the wells was carefully aspirated, after centrifugation was required for suspension cells, and the culture supernatant in the wells was aspirated. Add 150. Mu.l DMSO/well and shake for 10 minutes to dissolve the crystals well. And selecting a 490nm wavelength, measuring the light absorption value of each pore on an enzyme-linked immunosorbent assay, recording the result, and drawing a HaCat cell growth histogram by taking the concentration as an abscissa and the cell survival rate as an ordinate. The results are shown in FIG. 5;
the results show that: 0.5-2 mmol/L of EDTA, 0.05-0.15 wt% of polyhexamethylene biguanide salt and 0.05-0.15 wt% of undecylenamidopropyl betaine have no or less killing and inhibiting effects on normal cells.
4. Cell scratch test
The method comprises the steps of inoculating HaCaT cells to a 6-hole culture plate, scratching the cells after the cells adhere to the wall by using a sterilized 20uL gun head compared with a ruler, removing culture solution, washing for 3 times by using PBS (phosphate buffer solution), taking the embodiment 10 and the embodiment 1 as experimental groups, and taking normal saline as a control group. The culture solution 500uL,0, 24 and 48h are respectively added to observe the migration condition of the cells and take pictures under a microscope, and the percentage of the area of the coverage area of the migration of the cells of the experimental group in the initial scratch area is calculated by using Image J software. 3 sets of 3 wells each, and the experiment was repeated 3 times. The results are shown in FIG. 6;
the results show that: the combination of example 1 and example 10, compared to normal saline, showed a faster growth rate for normal cells; is beneficial to the growth of cells at the wound.
5. Determination of wound healing Rate
8 mice were simply divided into groups, 2 mice were blank control groups, and another 6 mice were divided into 3 groups as drug groups, and the mice were weighed and recorded before the start of the experiment, and 3% of the body weight (50 mg/kg) of the pentobarbital sodium mice were subjected to intraperitoneal injection for anesthesia treatment. After anesthesia, the back of the mouse is preliminarily shaved by using a mouse shaver, the shaving range of the back of the mouse is as large as possible, then 8% sodium sulfide (8 g of sodium sulfide nonahydrate, 7g of starch, 2g of glucose, 5g of glycerol, 1g of borax and 60ml of water) is used for thoroughly unhairing, and the skin on the back of the mouse is cleaned and wiped dry by using physiological saline, so that the steps are convenient for subsequent wound treatment. After the depilating treatment was completed, the mouse was properly fixed on the small animal test bed with the back facing upward, and the operation was prepared. Then, a skin wound of about 1.0cm × 1.0cm was made on the back of the mouse without damaging the fascia layer. Wound dressing concentration of 10 8 And slightly and uniformly coating the CFU/mL and 100 mu L of escherichia coli liquid by using a gun head, independently placing each mouse in an independent ventilating rearing cage after the mice are anesthetized and revived, infecting the wound by using the gun head after 24 hours, observing and taking a photograph of the infection change condition of the wound and the like until an obvious infection focus at the wound is formed.
The wound was changed daily with 10ml of irrigation solution each time, starting from the time of wound production, and wound healing was observed. On days 0, 2, 8, 12, 14, wound healing was observed and photographed, and the results are shown in fig. 7. And calculating the wound healing area of the mouse by using Image J software, and then counting the wound healing rate.
Table 7: statistics of ulcer area healing rate of grouped mice
From the data in table 7 it can be seen that:
in comparative example 1, the irrigating solution was 0.9% physiological saline, which has a disadvantage of low wound healing rate;
in comparative example 2, the concentration ratio of sodium, potassium, magnesium, calcium and the like is not proper, so that the osmotic pressure exceeds the cell tolerance range;
the healing rate of example 1 was higher than 0.9% saline (comparative example 1);
the healing rates of example 10 and example 14 were both higher than 0.9% saline (comparative example 1); in the embodiment 10 and the embodiment 14, on the basis of the embodiment 1, bacteriostatic components of 0.5-2 mmol/L of EDTA, 0.05-0.15 wt% of polyhexamethylene biguanide salt and 0.05-0.15 wt% of undecylenamidopropyl betaine are added and are matched in a synergistic manner, so that pathogenic microorganisms can be killed specifically, wound infection is prevented remarkably, and the wound healing rate is further improved.
In summary, one or more technical solutions of the embodiments of the present invention have at least the following advantages:
the flushing liquid special for pets of the invention has the components of 120-180 mmol/L of sodium ions, 5-15 mmol/L of potassium ions, 0.5-3 mmol/L of magnesium ions, 1-5 mmol/L of calcium ions, 10-40 mmol/L of acetate ions and 1-10 mmol/L of citrate ions which are most close to extracellular fluid components, has the minimum interference on the environment in body fluid of an organism, has the functions of maintaining the osmotic pressure of cells and regulating and controlling the balance of the pH value of cell sap, also has the function of bacteriostasis, is suitable for the treatment of wounds and infected wounds of common pets (including but not limited to cats and dogs), promotes the growth of wound cells, and is beneficial to the rapid healing of operation wounds.
According to the special pet washing liquid, 0.5-2 mmol/L of EDTA, 0.05-0.15 wt% of polyhexamethylene biguanide salt and 0.05-0.15 wt% of undecylenic amide propyl betaine are used as bacteriostatic components in a synergistic manner, so that pathogenic microorganisms can be specifically killed, wound infection is remarkably prevented, and the wound healing rate is further improved.
Finally, it should also be noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (8)
1. The special pet washing liquid is characterized by consisting of the following components:
sodium chloride, potassium chloride, calcium chloride, magnesium chloride, sodium acetate, sodium citrate, disodium EDTA, polyhexamethylene biguanide salt, and undecylenamidopropyl betaine; the following concentrations were used:
120 to 180mmol/L of sodium ions, 5 to 15mmol/L of potassium ions, 0.5 to 3mmol/L of magnesium ions, 1 to 5mmol/L of calcium ions, 10 to 40mmol/L of acetate ions, 1 to 10mmol/L of citrate ions, 1 to 10mmol/L of EDTA, 0.05 to 0.15wt percent of polyhexamethylene biguanide salt and 0.05 to 0.15wt percent of undecenylamidopropyl betaine; the pH value of the flushing liquid special for the pet is 6.8-8.0, and the osmotic pressure value is 260-350 mOsm/kg.
2. The pet rinse solution of claim 1, wherein the pet rinse solution is composed of, in concentration: 140-160 mmol/L of sodium ions, 8-12 mmol/L of potassium ions, 1.0-2.0 mmol/L of magnesium ions, 2-4 mmol/L of calcium ions, 15-35 mmol/L of acetate ions and 4-7 mmol/L of citrate ions.
3. The pet rinse solution of claim 1, wherein the pet rinse solution is composed of, in concentration: 154.2mmol/L of sodium ions, 10mmol/L of potassium ions, 1.48mmol/L of magnesium ions, 3.2mmol/L of calcium ions, 28.8mmol/L of acetate and 5.6mmol/L of citrate ions.
4. The rinse solution special for pets according to any of claims 1 to 3, wherein the EDTA is 2 to 8mmol/L, polyhexamethylene biguanide salt is 0.08 to 0.12wt% and undecenamidopropyl betaine is 0.08 to 0.12wt%.
5. The rinse solution for pet use according to any one of claims 1 to 3, wherein the EDTA is 5mmol/L, polyhexamethylene biguanide salt is 0.1wt% and undecylenamidopropylbetaine is 0.1wt%.
6. A method for preparing a pet-specific washing solution according to any one of claims 1 to 5, wherein the method comprises:
dissolving the components of the special pet washing liquid as claimed in any one of claims 1 to 5, sodium chloride, potassium chloride, calcium chloride, magnesium chloride, sodium acetate, sodium citrate, disodium EDTA, polyhexamethylene biguanide salt and undecenylamidopropylbetaine in purified water, adjusting the pH value of the solution, and sterilizing to obtain the special pet washing liquid.
7. The method of manufacturing of claim 6, wherein the sterilizing comprises: moist heat sterilization at 121 deg.C for 30min.
8. Use of a dedicated irrigating solution for pets according to any of claims 1 to 5 for the preparation of a medicament for wound irrigation of pets, including general wounds and/or infected wounds.
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