CN106377763B - Multi-purpose sterilizing and cleaning spray and its application - Google Patents
Multi-purpose sterilizing and cleaning spray and its application Download PDFInfo
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- CN106377763B CN106377763B CN201610884076.4A CN201610884076A CN106377763B CN 106377763 B CN106377763 B CN 106377763B CN 201610884076 A CN201610884076 A CN 201610884076A CN 106377763 B CN106377763 B CN 106377763B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/40—Transferrins, e.g. lactoferrins, ovotransferrins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/14—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12
- A23B4/18—Preserving with chemicals not covered by groups A23B4/02 or A23B4/12 in the form of liquids or solids
- A23B4/20—Organic compounds; Microorganisms; Enzymes
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- 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
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
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- 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
- A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
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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
- A61K9/10—Dispersions; Emulsions
- A61K9/12—Aerosols; Foams
Abstract
A multi-purpose sterilizing and cleaning spray comprises a base material liquid, wherein the base material liquid is a base material liquid formed by mixing bovine lactoferrin and water, or a base material liquid formed by mixing a combination of bovine lactoferrin, a hydrolysate or an zymolyte of bovine lactoferrin and water; when the base material liquid is selected as the former, the bovine lactoferrin accounts for 0.5-25% of the weight of the base material liquid; when the base material liquid is the latter, the sum of the weight of the bovine lactoferrin and the hydrolysate or zymolyte of the bovine lactoferrin accounts for 0.5-25% of the weight of the base material liquid, and the weight ratio of the bovine lactoferrin to the hydrolysate or zymolyte of the bovine lactoferrin is 100: 5-380; the hydrolysate or hydrolysate of bovine lactoferrin comprises one or a mixture of lactoferricin B, lactofettampin. The sterilization cleaning spray has broad-spectrum sterilization, can be sprayed on a human body without side effect or drug resistance, has good treatment effect, good taste and fragrance, good patient compliance and convenient carrying and use by colleagues; can be sprayed on fresh meat, and has good fresh-keeping effect, safety and harmlessness.
Description
Technical Field
The invention relates to the technical field of sterilization cleaning agents, in particular to a formula and a method of sterilization cleaning spray type medicines and cleaning agents which take bovine lactoferrin and hydrolysate or zymolyte thereof, namely lactoferricin B (LFcin B) and lactofettmampin (LFampin), and other peptides as main effective components.
Background
The environment in which people live is full of various bacteria, viruses, harmful dust and chemicals which easily cause harm to human bodies, and the harm to human bodies, particularly to sores and wounds on oral cavities, nasal cavities, throats, pudendum, nipples and other parts of the human bodies is larger. How to effectively protect and treat these vulnerable sites is a great concern. The following problems should be solved: the medicament selected by the treatment scheme is one, and has broad spectrum effect on viruses and bacteria; secondly, no side effect or few side effects; thirdly, drug resistance cannot be generated; fourthly, the taste and the fragrance are better, and the compliance of patients is good; fifth, the carrying and the use are convenient, and so on, and no ideal product is found yet. In addition, fresh-keeping agents harmless to health are also needed for fresh-keeping of fresh meat such as beef, sheep, pigs, ducks, chickens and the like.
A great deal of research at home and abroad shows that the bovine lactoferrin is a multifunctional bioactive protein, has good biological activity against various microorganisms (bacteria, viruses, fungi and the like), is a natural broad-spectrum antibiotic, has immunoregulatory activity on organisms and can enhance the immune function of the organisms. Bovine lactoferrin has a high affinity for binding iron and can compete with bacteria for iron ions in the internal environment of the body. Iron is an essential nutrient for the growth of pathogenic bacteria. The bovine lactoferrin has the functions of robbing and blocking bacteria from obtaining iron, so that bacteria, viruses, fungi and the like can be inactivated or can not grow and reproduce because sufficient iron ions can not be obtained. Bovine lactoferrin can cause detrimental changes in the structure of bacterial cell surface membranes. Such as bovine lactoferrin, can be released from the outer membrane of gram-negative bacteria by binding to Lipopolysaccharide (LPS) in the outer membrane, resulting in damage to the cell wall. In addition, bovine lactoferrin can also interact with escherichia coli, outer membrane porin (porin) of salmonella typhimurium, colonization factor (collagen factor) of haemophilus influenzae, and hemoglobin receptor of porphyromonas gingivalis to cause bacterial cell damage. It was reported by Marieke i.a (2005) that bovine lactoferrin contains two antimicrobial domain regions, called lactoferricin B (LFcin B) and lactofettampin (lfampin), respectively. The authors investigated the effect of these two typical antimicrobial peptide fragments on candida albicans and escherichia coli. Confocal laser scanning microscopy showed that these peptides could undergo cellular internalization within minutes, while cryo-etching of the transmission electron microscopy indicated that the integrity of the membrane was also compromised. The most striking findings are the unique vesicle-like structure produced on the Candida albicans membrane induced by Lfampin, and the outer membrane separation and surface elevation [ [ i ] ] of Escherichia coli caused by LFcin B peptide. In China, plum crystal and the like (2005) report that [ [ ii ] ], the multifunctional activity of bovine lactoferrin such as antibacterial and anti-inflammatory activity is related to the highly basic N-terminal region and can be released under the condition of acidic pH value, and the newly generated antimicrobial peptide containing 25 amino acid residues is called as lactoferrin. It readily reacts with negatively charged bacterial membrane structures and peptide binding can disrupt the normal structure of the membrane. Among the most important amino acid residues associated with the activity of lactoferricin are tryptophan and arginine. The bovine lactoferrin has protective and repairing effects on mucosa. Bovine lactoferrin has immunomodulatory activity and can promote the growth and differentiation of T lymphocytes. It has been reported abroad that lactoferrin uniquely binds to regions of Major Histocompatibility (MHC) proteins, as well as to the CD4 and CD8 determinants present on the surface of T4 (helper T cells) and T8 (suppressor T cells), and carries sequences homologous to MHC class II determinants. Lactoferrin also has the effect of regulating cytokines and lymphokines. Oral administration of bovine lactoferrin increased the number of NK cells (dose-dependent) in peripheral blood and spleen and increased the production of interferon-alpha, as recently reported by foreign scholars Kuhara (2006) [ [ iii ] ]. Experiments have also shown that oral administration of bovine lactoferrin also results in an immediate increase in IL-18 levels in the portal circulation. Furthermore, oral administration of bovine lactoferrin also increased the expression levels of interferon-alpha and interferon-beta in Peyer's Patches (PP) and Mesenteric Lymph Nodes (MLN). In summary, the above results demonstrate that oral administration of bovine lactoferrin stimulates gut-associated immune functions, including production of IL-18 and type I interferons, as well as potentiating NK cell activity. A large number of researches show that the lactoferrin with different iron saturation degrees has different bacteriostatic effects, and the verified that the iron saturation degree of the lactoferrin is in the range of 6-20%, and the bacteriostatic activity of the lactoferrin is enhanced along with the increase of the saturation degree. In addition, a large number of studies at home and abroad also show that peptides such as bovine lactoferrin and hydrolysate or zymolyte thereof, lactoferricin B (LFcin B), lactofettmampin (LFampin) and the like have the broad-spectrum bacteriostatic and bactericidal biological activity which is 200-fold and 400-fold higher than that of bovine lactoferrin.
Because of the above-mentioned characteristics of bovine lactoferrin and its hydrolysate or zymolyte, such as lactoferricin B (LFcin B) and lactofettmampin (LFampin), etc., it is a problem that needs to be solved at present if it is reasonably applied to realize better sterilization and cleaning functions.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a multi-purpose sterilization cleaning spray, which can be used for treating and maintaining sores at oral cavity, nasal cavity, throat, pudendum, nipple and other parts of human body, and can be used for keeping fresh of fresh meat of cattle, sheep, pig, duck, chicken and the like.
The multipurpose sterilizing clean-keeping spray comprises base material liquid, wherein the base material liquid is base material liquid formed by mixing bovine lactoferrin and water, or is base material liquid formed by mixing combination of bovine lactoferrin, hydrolysate or zymolyte of bovine lactoferrin and water; when the base material liquid is selected as the former, the bovine lactoferrin accounts for 0.5-25% of the weight of the base material liquid; when the base material liquid is the latter, the sum of the weight of the bovine lactoferrin and the hydrolysate or zymolyte of the bovine lactoferrin accounts for 0.5-25% of the weight of the base material liquid, and the weight ratio of the bovine lactoferrin to the hydrolysate or zymolyte of the bovine lactoferrin is 100: 5-380; the hydrolysate or zymolyte of the bovine lactoferrin comprises one or a mixture of lactoferricin B, lactofettampin.
Further, the spray also comprises one or the mixture of any of essence, monosodium glutamate and cooling agent; and are all added into the base material liquid to be mixed evenly.
Further, the hydrolysate or zymolyte molecular weight of the bovine lactoferrin is between 500-8500 Da.
Further, the hydrolysate or zymolyte of the bovine lactoferrin comprises lactoferrin B, and the weight ratio of the bovine lactoferrin to the lactoferrin B is 6-10: 0.5 to 2; the mass ratio of the sum of the weight of the bovine lactoferrin and the lactoferrin B to the water is 5-12: 100.
Further, the lactoferrin is low-iron saturation lactoferrin with iron saturation of 5-8%; the bovine lactoferrin, the hydrolysate or the zymolyte of the bovine lactoferrin are obtained by adding lactoferrin with low iron saturation into 1000u/g pepsin at constant temperature of 37 ℃ for enzymolysis for 15-25 min.
The preparation method of the low-iron saturation lactoferrin comprises the following steps:
step 1, adsorbing and analyzing lactoferrin in milk by using cation exchange resin; the analysis process adopts secondary analysis, the primary analysis adopts 1-2.5% of salt solution for analysis, and the analysis solution is lactoperoxidase protein and is discarded; the second-stage analysis is to adopt 3.5-7% of salt solution for analysis, and the analysis solution is sent to the step 2 for treatment; the salt solution is sodium salt, potassium salt or calcium salt solution;
step 2, concentrating the analytic solution obtained in the step 1 by adopting a separation membrane; the separation membrane is an ultrafiltration membrane with the molecular weight cutoff of 1000-30K;
step 3 and step 2, the concentrated solution obtained by the step adopts insoluble Fe3+Chelating resin is used for adsorption, and a separation membrane is used for concentrating the concentrated solution; during the adsorption operation, acetic acid, acetate and citrate are added into the concentrated solution to prepare the solution with the citrate concentration of 0.1-0.5 mol/L in the adsorption environment, and the pH value is adjusted to 5.0-6.7; the separation membrane is an ultrafiltration membrane with the cut-off molecular weight of 1000-30K.
Further, before the concentrated solution obtained in the step 2 is sent to the step 3, the concentrated solution can be subjected to adsorption and desorption of weak acid type cation resin and concentration treatment of an ultrafiltration membrane; the weak acid type cation resin is CM resin.
Further, in the step 3, the insoluble Fe3+The chelating resin is D403 chelating resin; the adsorption time of the resin is 20-120 minutes, the rotating speed rate of a stirring paddle during adsorption is 20-150 rpm, and the adsorption temperature is 3-30 ℃.
Further, in the step 1, the preparation step of the cation exchange resin is as follows:
step 1, taking 10g of chloracetyl polystyrene-divinylbenzene microsphere carrier PS-acyl-Cl, adding 60ml of tetrahydrofuran to swell for 12 hours, adding 40ml of methanol, and then adding the PS-acyl-Cl into the mixture according to the weight ratio: ethylene diamine: sodium bicarbonate is 1: 8: 0.7, sequentially adding ethylenediamine and sodium bicarbonate, stirring and reacting at 80 ℃ for 24 hours, after the reaction is finished, pouring the product into a sand core funnel, washing the product to be neutral by using distilled water, washing the product by using methanol, filtering the product for 3 times, and drying the product in vacuum until the weight is constant to obtain the cross-linked EDA microsphere carrier;
step 2, weighing 60g of cross-linked EDA microsphere carrier, placing the carrier in a three-neck flask, adding dimethylformamide to swell for 12 h, and then adding 16g of 2-acrylamide-2-methylpropanesulfonic acid, 3g of N-butoxy methacrylamide, dicyclopentenyl acrylate and K2CO35g and 12g of tetrabutylammonium bromide, stirring the mixture, performing reflux reaction in an oil bath at a certain temperature, transferring a reaction product into a sand core funnel after the reaction is finished, cleaning the reaction product for 4 times by using 5wt% of HC1, filtering and washing the reaction product to be neutral by using distilled water, finally washing and filtering the reaction product by using methanol, draining the washing product, and drying the washing product in vacuum to constant weight to obtain the cation exchange resin
The multi-purpose sterilizing and cleaning spray is applied to inhibiting and killing bacteria and viruses. For example, the spray is sprayed as mist or solution fine particles by pressing a gas valve with hand, and is used for treating and maintaining oral cavity, throat, nasal cavity, female vagina, vulva, nipple, male vulva, sore and wound under armpit, or for keeping fresh meat.
The sterilization cleaning spray has broad-spectrum sterilization, can be sprayed on a human body without side effect or drug resistance, has good treatment effect, good taste and fragrance, good patient compliance and convenient carrying and use by colleagues; can be sprayed on fresh meat, and has good fresh-keeping effect, safety and harmlessness.
Detailed Description
The invention will be further described with reference to specific examples, wherein the amounts of the flavors, pigments and sweeteners are all trace and can be adjusted slightly according to actual needs.
Example 1:
a multi-purpose sterilizing and cleaning spray comprises the following components:
lactoferrin with low iron saturation of 7g
Borneol 1.5g
Menthol 1g
Appropriate amount of sweetener
Right amount of olive pigment
Proper amount of essence
100ml of water
1. Preparation of low iron saturation lactoferrin: adding SPC70 resin into a stirring tank, adsorbing for 80min at the stirring speed of 60rpm, collecting the adsorbed milk, washing the resin with RO water, adding a first-stage 100L 2.5% NaCl aqueous solution, desorbing for 40min at the stirring speed of 40rpm, collecting the desorbed solution, and leaching and collecting with RO water; adding 100L of 5.5% NaCl aqueous solution of the second stage, desorbing for 40min, stirring at 40rpm, collecting the desorbed solution, and eluting with RO water to collect the resin for 3 times. The second-stage desorption solution and the second RO eluent are combined and collected for 180L, and conveyed to a microfiltration membrane device with the diameter of 0.1 μm for degerming and impurity removal; desalting and concentrating by using ultrafiltration membrane equipment with 30K molecular cut-off, and sampling to detect that the iron saturation is 20%; inputting the lactoferrin concentrated solution into a CM resin tank to remove impure proteins, collecting the lactoferrin solution adsorbed with the impure proteins, continuously performing desalination and concentration by using ultrafiltration membrane equipment with 10K molecular cut-off, and performing freeze drying on a small amount of concentrated solution to obtain light pink dry powder. The water content of the powder is 3.0 percent, the ash content is 1.1 percent, the total protein content is 96.3 percent, and the iron saturation is 20 percent; and inputting the residual lactoferrin concentrated solution into a tank filled with D403 chelating resin for deferrization, adding 0.1mol/L citrate, adjusting the pH value to 6.0 by using acetic acid-sodium acetate buffer solution, stirring to adsorb iron ions, wherein the resin adsorption time is 120 minutes, and the rotating speed of a stirring paddle is 60 rpm. Collecting the lactoferrin solution after adsorbing iron ions, continuously desalting and concentrating by using an ultrafiltration membrane device with 10K molecular cut-off, collecting the concentrated solution, and freeze-drying to obtain light pink dry powder. The powder contained 3.0% water, 1.0% ash, 96.3% total protein, and 7% iron saturation.
2. Adding the rest materials into lactoferrin with low iron saturation.
Example 2
A multi-purpose sterilizing and cleaning spray comprises the following components:
lactoferrin with low iron saturation of 7g
Borneol 1.5g
Menthol 1g
Appropriate amount of sweetener
Right amount of olive pigment
Proper amount of essence
100ml of water
1. Preparation of low iron saturation lactoferrin: adding SPC70 resin into a stirring tank, adsorbing for 80min at the stirring speed of 60rpm, collecting the adsorbed milk, washing the resin with RO water, adding a first-stage 100L 2.5% NaCl aqueous solution, desorbing for 40min at the stirring speed of 40rpm, collecting the desorbed solution, and leaching and collecting with RO water; adding 100L of 5.5% NaCl aqueous solution of the second stage, desorbing for 40min, stirring at 40rpm, collecting the desorbed solution, and eluting with RO water to collect the resin for 3 times. The second-stage desorption solution and the second RO eluent are combined and collected for 180L, and conveyed to a microfiltration membrane device with the diameter of 0.1 μm for degerming and impurity removal; desalting and concentrating by using ultrafiltration membrane equipment with 30K molecular cut-off, and sampling to detect that the iron saturation is 20%; inputting the lactoferrin concentrated solution into a CM resin tank to remove impure proteins, collecting the lactoferrin solution adsorbed with the impure proteins, continuously performing desalination and concentration by using ultrafiltration membrane equipment with 10K molecular cut-off, and performing freeze drying on a small amount of concentrated solution to obtain light pink dry powder. The water content of the powder is 3.0 percent, the ash content is 1.1 percent, the total protein content is 96.3 percent, and the iron saturation is 20 percent; and inputting the residual lactoferrin concentrated solution into a tank filled with D403 chelating resin for deferrization, adding 0.1mol/L citrate, adjusting the pH value to 6.0 by using acetic acid-sodium acetate buffer solution, stirring to adsorb iron ions, wherein the resin adsorption time is 120 minutes, and the rotating speed of a stirring paddle is 60 rpm. Collecting the lactoferrin solution after adsorbing iron ions, continuously desalting and concentrating by using an ultrafiltration membrane device with 10K molecular cut-off, collecting the concentrated solution, and freeze-drying to obtain light pink dry powder. The powder contained 3.0% water, 1.0% ash, 96.3% total protein, and 7% iron saturation.
2. Lactoferrin with low iron saturation is added into pepsin with the concentration of 800-.
3. Adding the rest materials into the mixture of lactoferrin and lactoferricin B.
Example 3
A multi-purpose sterilizing and cleaning spray comprises the following components:
lactoferrin 9g with low iron saturation
Borneol 2g
Menthol 1.5g
Appropriate amount of sweetener
Right amount of olive pigment
Proper amount of essence
100ml of water
1. Preparation of low iron saturation lactoferrin: adding SPC70 resin into a stirring tank, adsorbing for 80min at the stirring speed of 60rpm, collecting the adsorbed milk, washing the resin with RO water, adding a first-stage 100L 2.5% NaCl aqueous solution, desorbing for 40min at the stirring speed of 40rpm, collecting the desorbed solution, and leaching and collecting with RO water; adding 100L of 5.5% NaCl aqueous solution of the second stage, desorbing for 40min, stirring at 40rpm, collecting the desorbed solution, and eluting with RO water to collect the resin for 3 times. The second-stage desorption solution and the second RO eluent are combined and collected for 180L, and conveyed to a microfiltration membrane device with the diameter of 0.1 μm for degerming and impurity removal; desalting and concentrating by using ultrafiltration membrane equipment with 30K molecular cut-off, and sampling to detect that the iron saturation is 20%; inputting the lactoferrin concentrated solution into a CM resin tank to remove impure proteins, collecting the lactoferrin solution adsorbed with the impure proteins, continuously performing desalination and concentration by using ultrafiltration membrane equipment with 10K molecular cut-off, and performing freeze drying on a small amount of concentrated solution to obtain light pink dry powder. The water content of the powder is 3.0 percent, the ash content is 1.1 percent, the total protein content is 96.3 percent, and the iron saturation is 20 percent; and inputting the residual lactoferrin concentrated solution into a tank filled with D403 chelating resin for deferrization, adding 0.1mol/L citrate, adjusting the pH value to 6.0 by using acetic acid-sodium acetate buffer solution, stirring to adsorb iron ions, wherein the resin adsorption time is 120 minutes, and the rotating speed of a stirring paddle is 60 rpm. Collecting the lactoferrin solution after adsorbing iron ions, continuously desalting and concentrating by using an ultrafiltration membrane device with 10K molecular cut-off, collecting the concentrated solution, and freeze-drying to obtain light pink dry powder. The powder contained 3.0% water, 1.0% ash, 96.3% total protein, and 7% iron saturation.
2. Adding lactoferrin with low iron saturation into pepsin with the concentration of 800-: 1.
3. adding the rest materials into the mixture of lactoferrin and lactoferricin B.
Example 4
A multi-purpose germicidal sanitation spray in accordance with example 3, except that SPC70 resin was replaced with a cation exchange resin prepared by the following method:
step 1, taking 10g of chloracetyl polystyrene-divinylbenzene microsphere carrier (PS-acyl-Cl), adding 60ml of tetrahydrofuran to swell for 12 hours, adding 40ml of methanol, and then mixing according to the weight ratio (PS-acyl-Cl): ethylenediamine (EDA): sodium bicarbonate is 1: 8: 0.7, sequentially adding ethylenediamine and sodium bicarbonate, stirring and reacting at 80 ℃ for 24 hours, after the reaction is finished, pouring the product into a sand core funnel, washing the product to be neutral by using distilled water, washing the product by using methanol, filtering the product for 3 times, and drying the product in vacuum until the weight is constant to obtain the cross-linked EDA microsphere carrier;
step 2, weighing 60g of cross-linked EDA microsphere carrier, placing the carrier in a three-neck flask, adding dimethylformamide to swell for 12 h, and then adding 16g of 2-acrylamido-2-methylpropanesulfonic Acid (AMPS), 3g of N-butoxymethacrylamide (NBMA), dicyclopentenyl acrylate (DCPA) and K2CO3 5g and 12g of tetrabutylammonium bromide (TBAB), stirring, carrying out reflux reaction in an oil bath at a certain temperature, transferring a reaction product into a sand core funnel after the reaction is finished, washing for 4 times by using 5wt% of HC1, filtering and washing by using distilled water to be neutral, finally washing and filtering by using methanol, draining, and drying in vacuum to constant weight to obtain the cation exchange resin.
The technical indexes of the finally obtained low-iron saturation lactoferrin are as follows: the powder contained 3.0% water, 1.0% ash, 96.8% total protein and 6% iron saturation.
2. Adding lactoferrin with low iron saturation into pepsin with the concentration of 800-: 1.
3. adding the rest materials into the mixture of lactoferrin and lactoferricin B.
In vitro antibacterial test of multi-purpose bactericidal spray
In vitro antibacterial test, the concentration of the multi-purpose bactericidal spray prepared in example 2 is 5000mg/l, and the reference substance is Jieeryin lotion produced by Chengdu Enwei pharmaceutical Co Ltd; the test strains were: staphylococcus aureus ATCC 6538, candida albicans ATCC 10231, bacillus subtilis ATCC 6633. The test data are shown in the following table:
in vitro sterilization experiments show that: the multi-purpose sterilizing clean-keeping spray has strong sterilizing effect on staphylococcus aureus, the sterilizing rate can reach more than 99 percent when the spray is acted for 5 minutes under three concentrations (50 mg/l, 100mg/l and 200 mg/l); the bactericidal composition has a certain killing effect on bacillus subtilis, and the sterilization rate can reach more than 90% under three concentrations (50 mg/l, 100mg/l and 200 mg/l); has strong killing effect on candida albicans, acts for 10 minutes under three concentrations (50 mg/l, 100mg/l and 200 mg/l), and the sterilization rate can reach more than 98.5 percent on average.
Comparative tests are carried out on the first, second, third and fourth examples, and the experimental results are shown in the following table, compared with the best effect of example 4:
Claims (7)
1. a multi-purpose sterilizing and cleaning spray is characterized in that: the base liquid is base liquid formed by mixing bovine lactoferrin and water, or is base liquid formed by mixing the combination of bovine lactoferrin, hydrolysate or zymolyte of bovine lactoferrin and water; when the base material liquid is selected as the former, the bovine lactoferrin accounts for 0.5-25% of the weight of the base material liquid; when the base material liquid is the latter, the sum of the weight of the bovine lactoferrin and the hydrolysate or zymolyte of the bovine lactoferrin accounts for 0.5-25% of the weight of the base material liquid, and the weight ratio of the bovine lactoferrin to the hydrolysate or zymolyte of the bovine lactoferrin is 100: 5-380; the hydrolysate or zymolyte of the bovine lactoferrin comprises one or the mixture of lactoferricin B, lactofettampin;
the bovine lactoferrin is low-iron-saturation lactoferrin with iron saturation of 5-8%; the bovine lactoferrin, the hydrolysate or the zymolyte of the bovine lactoferrin are products obtained by adding lactoferrin with low iron saturation into pepsin with the temperature of 800-;
the preparation method of the low-iron saturation lactoferrin comprises the following steps:
step 1, adsorbing and analyzing lactoferrin in milk by using cation exchange resin; the analysis process adopts secondary analysis, the primary analysis adopts 1-2.5% of salt solution for analysis, and the analysis solution is lactoperoxidase protein and is discarded; the second-stage analysis is to adopt 3.5-7% of salt solution for analysis, and the analysis solution is sent to the step 2 for treatment; the salt solution is sodium salt, potassium salt or calcium salt solution;
step 2, concentrating the analytic solution obtained in the step 1 by adopting a separation membrane; the separation membrane is an ultrafiltration membrane with the molecular weight cutoff of 1000-30K;
step 3, adsorbing the concentrated solution obtained in the step 2 by using insoluble Fe3+ chelate resin, and concentrating the concentrated solution by using a separation membrane; during the adsorption operation, acetic acid, acetate and citrate are added into the concentrated solution to prepare the solution with the citrate concentration of 0.1-0.5 mol/L in the adsorption environment, and the pH value is adjusted to 5.0-6.7; the separation membrane is an ultrafiltration membrane with the molecular weight cutoff of 1000-30K;
the spray also comprises one or a mixture of any of essence, monosodium glutamate and algefacient, and is added into the base material liquid to be mixed uniformly.
2. The multi-purpose sterilizing and cleaning spray according to claim 1, characterized in that: the hydrolysate or zymolyte of the bovine lactoferrin has a molecular weight of between 500-8500 Da.
3. The multi-purpose sterilizing and cleaning spray according to claim 1, characterized in that: the hydrolysate or zymolyte of the bovine lactoferrin comprises lactoferrin B, and the weight ratio of the bovine lactoferrin to the lactoferrin B is 6-10: 0.5 to 2; the mass ratio of the sum of the weight of the bovine lactoferrin and the lactoferrin B to the water is 5-12: 100.
4. The multi-purpose sterilizing cleaning spray according to claim 1, wherein in the step 3, the insoluble Fe3+ chelating resin is D403 chelating resin; the adsorption time of the resin is 20-120 minutes, the rotating speed rate of a stirring paddle during adsorption is 20-150 rpm, and the adsorption temperature is 3-30 ℃.
5. The multi-purpose sterilizing and cleaning spray according to claim 1, wherein in the step 1, the cation exchange resin is prepared by the following steps:
step 1, taking 10g of chloracetyl polystyrene-divinylbenzene microsphere carrier PS-acyl-Cl, adding 60ml of tetrahydrofuran to swell for 12 hours, adding 40ml of methanol, and then adding the PS-acyl-Cl into the mixture according to the weight ratio: ethylene diamine: sodium bicarbonate is 1: 8: 0.7, sequentially adding ethylenediamine and sodium bicarbonate, stirring and reacting at 80 ℃ for 24 hours, after the reaction is finished, pouring the product into a sand core funnel, washing the product to be neutral by using distilled water, washing the product by using methanol, filtering the product for 3 times, and drying the product in vacuum until the weight is constant to obtain the cross-linked EDA microsphere carrier;
step 2, weighing 60g of a cross-linked EDA microsphere carrier, placing the carrier in a three-necked flask, adding dimethylformamide to swell for 12 h, adding 16g of 2-acrylamido-2-methylpropanesulfonic acid, 3g of N-butoxymethacrylamide, dicyclopentenyl acrylate, K2CO 35 g and 12g of tetrabutylammonium bromide, stirring the mixture to perform reflux reaction in an oil bath at a certain temperature, transferring a reaction product to a sand core funnel after the reaction is finished, cleaning the reaction product for 4 times by using 5wt% HC1, filtering and washing the reaction product to be neutral by using distilled water, finally washing and filtering the reaction product by using methanol, draining the reaction product, and drying the reaction product in vacuum to constant weight to obtain the cation exchange resin.
6. The use of the multi-purpose germicidal and sanitation spray as claimed in any one of claims 1 to 5 in the preparation of a medicament for inhibiting and killing bacteria and viruses.
7. The use of the multi-purpose sterilizing cleaning spray according to any one of claims 1-5 in fresh-keeping of raw meat.
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|---|---|---|---|---|
| CN1840179A (en) * | 2006-01-19 | 2006-10-04 | 方雅悯 | Application of bovine lactoferricin in preparation of medicine for treating HP infected gastric disease |
| CN101247823A (en) * | 2005-05-05 | 2008-08-20 | 布里斯托尔-迈尔斯斯奎布公司 | Use of bovine lactoferrin in the manufacture of a medicament for inhibiting the growth of bacteria |
| CN105999236A (en) * | 2016-07-06 | 2016-10-12 | 方雅悯 | Medicament for treating bovine mastitis and application thereof |
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2016
- 2016-10-11 CN CN201610884076.4A patent/CN106377763B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101247823A (en) * | 2005-05-05 | 2008-08-20 | 布里斯托尔-迈尔斯斯奎布公司 | Use of bovine lactoferrin in the manufacture of a medicament for inhibiting the growth of bacteria |
| CN1840179A (en) * | 2006-01-19 | 2006-10-04 | 方雅悯 | Application of bovine lactoferricin in preparation of medicine for treating HP infected gastric disease |
| CN105999236A (en) * | 2016-07-06 | 2016-10-12 | 方雅悯 | Medicament for treating bovine mastitis and application thereof |
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| Ultrastructural effects of antimicrobial peptides from bovine lactoferrin on the membranes of Candida albicans and Escherichia coli;Kraan, M. I. A. van d等;《Peptides》;20050930;第90卷(第3期);第1537-1542页 * |
| 酶解牛乳铁蛋白制备乳铁素B的酶解条件研究;张东送等;《中国乳品工业》;20040531;第32卷(第5期);第13-15页 * |
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