CN104042719B - A kind of antibacterial modified chitosan spray and preparation method thereof - Google Patents
A kind of antibacterial modified chitosan spray and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to a kind of antibacterial modified chitosan spray and preparation method thereof.This preparation method comprises and prepares modification of chitosan, prepare Fructus Gardeniae oil, prepare jasminoidin, prepare spirulina polysaccharide and the step such as spirulina polypeptide and preparation chitosan spray.Spray of the present invention is applied to treatment men and women's property pruritus vulvae and fungal infection, courses of infection, folliculitis, vaginitis, also be applied to the treatment etc. of decubital ulcer and skin Chronic inflammation, application is very extensive, and it reaches more than 99% to the antibacterial ability of staphylococcus aureus, escherichia coli, streptococcus, Candida albicans, bacillus pyocyaneus or other fungus.
Description
[technical field]
The invention belongs to medical drugs technical field.More specifically, the present invention relates to a kind of antibacterial modified chitosan spray, also relate to the preparation method of described antibacterial modified chitosan spray,
[background technology]
Chitosan is that one derives from the shell of Crustacean (Eriocheir sinensis shrimp) and the natural alkaline macromolecule polysaccharide of segmental appendage class animal (Blatta seu periplaneta etc.) skeleton, there is good biocompatibility, biodegradability and regeneration that is antibacterial, anticorrosion, that promote endotheliocyte, blood capillary, myocardial cell and neural axon, the premium properties such as hemostasis and promotion wound healing.Research shows, low-molecular weight chitoglycan has human body and easily absorbs, can the physiologically active such as activated lymphocyte, enhancing human body immunity power, antibacterial bacteriostatic, inhibition tumor cell growth, in addition, research finds, deacetylating degree of chitosan is higher, and the effect of its scavenging hydroxyl, superoxide radical and organic free radical is better.These character make it have good application prospect as natural antibacterial, anti-biotic material, antiseptic.
Along with automiser spray is gradually improved, ejecting agent continues to bring out, for adapting to the needs of the allegro life style of modern society and clinical practice, the preparation technology of various spray is tending towards ripe, in local, whole body therapeutic and environment disinfected etc., application is more extensive, the advantage of abundant its uniqueness of display, development in recent years is very fast, has become an important branch in current medical instruments field.
Spray solely shows advantage in topical therapeutic, because medicinal liquid can be uniformly distributed in local patient part, and can absorb rapidly and play a role.At present, at treatment burn, scald Be very effective, cure rate is all more than 90%.Clinical demand in infantile eczema, tinea versicolor, closed bone injuries, beriberi and various dermatitis, gynaecopathia, skin injury and ulcer in body surface, periarthritis, control scar hyperplasia etc. is in rising trend.
At present, people utilize chitosan to prepare spray, such as CN102846655A discloses a kind of antibacterial spray and preparation method thereof, this spray is made up of water, chitosan, glycerol, polyhexamethylene guanidine, vitamin E, wherein 0.01% chitosan is difficult to play antibacterial effect, antibacterial main body polyhexamethylene guanidine is a kind of high molecular polymer of synthesis, is mainly used in aquatic products because of bacterial various prevention and treatment of diseases.CN102860567A discloses the fresh-keeping spray of a kind of chitosan, it by, chitosan, edible glue, glacial acetic acid, citric acid and water makes, but it does not clearly demonstrate molecular weight and the deacetylation of chitosan, does not evaluate its antibacterial, corrosion-resistanting fresh-keeping effect yet.CN102524316B discloses a kind of chitosan-based anti-freezing antibacterial spraying agent for cutting surfaces of rubber trees and preparation method thereof, this spray contains chitosan, tea tree ethereal oil, polyvinyl alcohol, plant growth regulator etc., it does not clearly demonstrate molecular weight and the deacetylation of chitosan equally, does not evaluate its antibacterial effect yet.
In addition, have not yet to see about utilizing chitosan to prepare the patent of spray as primary raw material.The chitosan of current application is mainly polymer chitosan, does not also see by chitin modified, controls its molecular weight and deacetylation to improve the relevant report of spray antibacterial action.
The object of the invention is to overcome the shortcoming existed in prior art, solve problems such as lacking the environmental protection of pure chitosan class, high-efficiency antimicrobial spray at present, a kind of preparation method of antibacterial modified chitosan spray is proposed, it is good that the spray obtained has spray effect, medicinal liquid can be uniformly distributed in patient part, and can absorb rapidly and play a role, antibacterial wide spectrum, can promote that focus cytothesis grows with healing simultaneously, can be applicable to the local of sufferer, whole body therapeutic.
[summary of the invention]
[technical problem that will solve]
The object of this invention is to provide a kind of antibacterial modified chitosan spray.
Another object of the present invention is to provide the preparation method of described antibacterial modified chitosan spray.
[technical scheme]
The present invention is achieved through the following technical solutions.
The present invention relates to a kind of preparation method of antibacterial modified chitosan spray.
The step of this antibacterial modified chitosan spray preparation method is as follows:
A, prepare modification of chitosan
By deacetylation 80%, molecular weight 20 × 10
4~ 30 × 10
4the chitosan of Da is dissolved in by weight in 1.0 ~ 3.0% aqueous acetic acids, be incorporated as two t-amyl peroxy compounds of chitosan weight 1 ~ 3 times again, the solution obtained carries out supersound process in ultrasonic washing unit, obtain low-molecular weight chitoglycan complex solution, again through polyacrylonitrile ultrafiltration film molecular retention and classification, obtain the chitosan that molecular weight is below 500Da; This chitosan is added to by weight in 35 ~ 45% potassium hydroxide aqueous solutions, 10 ~ 20min is processed under the condition of temperature 105 ~ 115 DEG C, then with 1.0 ~ 3.0N aqueous hydrochloric acid solution carry out neutralizing, filter, distilled water wash, drying, obtain deacetylation 99 ~ 100%, molecular weight be the modification of chitosan of below 500Da;
B, prepare Fructus Gardeniae oil
700 ~ 900 weight portion petroleum hydrocarbons are added in 80 ~ 120 weight portion gardenia powder, then in ultrasonic washing unit, supersound extraction is carried out, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 2 ~ 3 times in the same fashion with 500 ~ 700 weight portion petroleum ether, filtrate merges, employing heating in water bath mode removes the petroleum hydrocarbon in merging filtrate under the condition of temperature 60 ~ 90 DEG C, obtains Fructus Gardeniae oil;
C, prepare jasminoidin
800 ~ 1200 weight portions, 70 ~ 80% ethanol waters are by volume added in 80 ~ 120 weight portion gardenia powder, then in ultrasonic washing unit, supersound extraction is carried out, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 2 ~ 3 times in the same fashion, filtrate merges, then at flow velocity be under the condition of 20 ~ 30 microlitres/second by nonpolarity macroporous adsorptive resins chromatographic column and at flow velocity be 30 ~ 37 microlitres/second condition under pass through polyamide column, then working concentration 15 ~ 25% ethanol water eluting agent are by volume from adsorption column eluting, the eluent obtained processes in a rotary evaporator and obtains jasminoidin under the condition of distilling under reduced pressure,
D, prepare spirulina polysaccharide and spirulina polypeptide
Fresh spirulina is processed 12 ~ 18 minutes in low temperature biomixer under 4 DEG C ~ 6 DEG C cryogenic conditions, the treatment fluid obtained carries out centrifugal treating through low temperature supercentrifuge again under temperature 4 DEG C ~ 6 DEG C conditions, then filter at low temperature equipment is used to filter under the condition of temperature 2 DEG C ~ 5 DEG C, then the permeable membrane using molecular cut off to be 10kD under the condition of temperature 2 DEG C ~ 8 DEG C carries out membrane permeation process, obtains spirulina polysaccharide and spirulina polypeptide;
E, preparation chitosan spray
2.5 ~ 3.2 weight portions are dissolved in 6 ~ 15 weight portion concentration by weight in 0.8 ~ 1.2% aqueous acetic acid at the modification of chitosan that steps A obtains, then in obtained solution, 0.8 ~ 1.2 weight portion polysorbate60 is added, 0.8 ~ 1.2 weight portion lauroylamidopropyl betaine, mix homogeneously, add the jasminoidin that 0.3 ~ 0.8 weight portion obtains in step C again, the Fructus Gardeniae oil that 0.1 ~ 0.4 weight portion obtains in step B, the spirulina polypeptide that 0.1 ~ 0.4 weight portion obtains in step D, the spirulina polysaccharide that 0.1 ~ 0.4 weight portion obtains in step D, mix homogeneously, finally add 0.1 ~ 0.4 weight portion pi-allyl tributyl bromide phosphine, with distilled water, its gross weight is reached 100 weight portions, obtain described antibacterial modified chitosan spray.
A preferred embodiment of the invention, in step, dissolves 5 ~ 15min under the condition of deacetylation 80% chitosan low whipping speed 250 ~ 350rpm in aqueous acetic acid.
According to another kind of preferred implementation of the present invention, in step, the solution obtained supersound process 35 ~ 45min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W.
According to another kind of preferred implementation of the present invention, in stepb, described filtering residue is when the 1st supersound extraction, and the weight ratio of gardenia powder and petroleum hydrocarbon is 90 ~ 110:750 ~ 850; When the 2nd time or the 3rd supersound extraction, the weight ratio of gardenia powder and petroleum hydrocarbon is 90 ~ 110:550 ~ 650.
According to another kind of preferred implementation of the present invention, in step C, described macroporous adsorbent resin is nonpolar D101, LX-60 or LX-20 type resin prepared by styrene and divinylbenzene combined polymerization; Described polyamide synthesizes by caprolactam or adipic acid and hexamethylene diamine the high-molecular polyamide prepared.
According to another kind of preferred implementation of the present invention, in step C, the eluant of described macroporous adsorptive resins or the eluant of described polyamide column are concentration 18 ~ 22% ethanol waters by volume.
According to another kind of preferred implementation of the present invention, in step D, described permeable membrane is the permeable membrane prepared by polyether sulfone, Triafol T or regenerated cellulose.
According to another kind of preferred implementation of the present invention, in step D, the rotating speed of described low temperature supercentrifuge is 20000 ~ 30000 revs/min.
According to another kind of preferred implementation of the present invention, the weight ratio of modification of chitosan, polysorbate60, lauroylamidopropyl betaine, jasminoidin, Fructus Gardeniae oil, spirulina polypeptide, spirulina polysaccharide and pi-allyl tributyl bromide phosphine is 2.8 ~ 3.0:0.85 ~ 1.12:0.85 ~ 1.12:0.45 ~ 0.65:0.15 ~ 0.24:0.15 ~ 0.26:0.15 ~ 0.25:0.15 ~ 0.24.
The invention still further relates to the antibacterial modified chitosan spray adopting described preparation method to obtain.The antibacterial ability of described antibacterial modified chitosan spray to staphylococcus aureus, escherichia coli, streptococcus, Candida albicans, bacillus pyocyaneus or other fungus reaches more than 99%.
In more detail the present invention will be described below
The present invention relates to a kind of preparation method of antibacterial modified chitosan spray.
The present invention with pure-natural biological mass shell polysaccharide for main matrix, by adjusting its molecular weight, change its deacetylation, again with other synergist with the use of, be aided with the combination of the nutrition and growth regulating adjuvant etc. that can promote that histiocyte is repaired simultaneously, prepare novel chitosan anti-bacteria spray, in the treatment for the treatment of gynaecopathia, skin injury and ulcer in body surface, men and women's property pudendal pruritus, infantile eczema, tinea versicolor, beriberi and various dermatitis etc., solely show advantage.
The step of modified chitosan anti-bacteria spray preparation method of the present invention is as follows:
A, prepare modification of chitosan
By deacetylation 80%, molecular weight 20 × 10
4~ 30 × 10
4the chitosan of Da is dissolved in by weight in 1.0 ~ 3.0% aqueous acetic acids, be incorporated as two t-amyl peroxy compounds of chitosan weight 1 ~ 3 times again, the solution obtained carries out supersound process in ultrasonic washing unit, obtain low-molecular weight chitoglycan complex solution, again through polyacrylonitrile ultrafiltration film molecular retention and classification, obtain the chitosan that molecular weight is below 500Da; This chitosan is added to by weight in 35 ~ 45% potassium hydroxide aqueous solutions, 10 ~ 20min is processed under the condition of temperature 105 ~ 115 DEG C, then with 1.0 ~ 3.0N aqueous hydrochloric acid solution carry out neutralizing, filter, distilled water wash, drying, obtain deacetylation 99 ~ 100%, molecular weight be the modification of chitosan of below 500Da.
The chemical constitution of chitosan (β-(1 → 4)-2-amino-2-deoxy-D-Glucose) is band cationic macromolecule alkalescence polysaccharide polymer, is nature uniquely positively charged, cation edible fiber; It has the physicochemical properties such as stable physical property, good dissolubility, hydrophilic hygroscopicity, biological degradability, biocompatibility and film property, caking property, breathability.
In step, 5 ~ 15min is dissolved under the condition of deacetylation 80% chitosan low whipping speed 250 ~ 350rpm in aqueous acetic acid.
The chitosan that the present invention uses is product sold in the market, such as by Jinan Haidebei Marine Organism Engineering Co., Ltd. with trade name food pharmaceutical grade chitosan product sold.
The effect adding two t-amyl peroxy compounds (Di-tert-amylperoxide) in this step makes chitosan molecule chain produce fracture by the effect of peroxide, form low molecular chitosan, the object using organic peroxide instead of employing hydrogen peroxide makes chain breaking process gentleness controlled, and can not produce uncontrollable chain-breaking reaction.
The two t-amyl peroxy compounds that the present invention uses are product solds in the market, such as by Taizhou City Hai Xiang Chemical Co., Ltd. with trade name t-amyl peroxy compound product sold.
In the present invention, the object of ultrasonic waves for cleaning process is to allow high molecular weight chitosan aqueous acetic acid carry out ultrasonic Treatment 30 ~ 50 minutes under the condition of ultrasonic frequency 20 ~ 25kHz and power 180 ~ 220W, obtains low-molecular weight chitoglycan complex solution.Preferably, high molecular weight chitosan aqueous acetic acid supersound process 35 ~ 45min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W.
The ultrasonic washing unit that the present invention uses is product sold in the market, such as by. Changzhou You Da ultrasonic equipment company limited is with trade name YD1008 ultrasonic washing unit product sold.
The present invention is by molecule hyperfiltration technique; by chitosan classification; intercept the chitosan with best molecular level scope that molecular weight is less than 500Da; then by again deacetylated; obtain high deacetylized, low-molecular-weight, narrow molecular weight distribution modification of chitosan, greatly improve broad-spectrum antibacterial property energy.
Normally a kind of hydrophobicity ultrafilter membrane with good any biological inert, chemical stability, heat stability of polyacrylonitrile ultrafiltration film.
Carrying out classification with polyacrylonitrile ultrafiltration film is that a kind of chitosan molecule molecular weight being greater than 500Da retains, and the operation that the chitosan molecule allowing molecular weight be less than 500Da is passed through.The molecular weight of chitosan adopts conventional Gel Permeation chromatographic process to measure and obtains.
The performance of the polyacrylonitrile ultrafiltration film that the present invention uses is as follows: thickness 50-90 μm, Unit Weight 2.1-3.4mg/cm
2, porosity 75-81%, intensity is greater than 15pst.
The polyacrylonitrile ultrafiltration film that the present invention uses is product sold in the market, such as by Zhaoyuan Jinhuimo Technology Co., Ltd. with trade name UFbIA250-PAN film product sold.
In a fractionated, the molecular weight collected is allowed to be that the chitosan molecule part of below 500Da is dried to water content lower than 6% in hot-air dry equipment under the condition of temperature 60 C.
Then, modification is carried out to the chitosan molecule alkali that obtained molecular weight is below 500Da, i.e. further deacetylated process.
In this modification procedure, the object of washing is salt and other impurity of removing neutralization generation.Dry object is to remove moisture, and the equipment that this drying steps uses is identical with previously described, and baking temperature condition is also identical with previously described.
B, prepare Fructus Gardeniae oil
700 ~ 900 weight portion petroleum hydrocarbons are added in 80 ~ 120 weight portion gardenia powder, then in ultrasonic washing unit, supersound extraction is carried out, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 2 ~ 3 times in the same fashion with 500 ~ 700 weight portion petroleum ether, filtrate merges, employing heating in water bath mode removes the petroleum hydrocarbon in merging filtrate under the condition of temperature 60 ~ 90 DEG C, obtains Fructus Gardeniae oil;
Gardenia powder is product sold in the market, such as by north and south, Guangzhou row prepared slices of Chinese crude drugs company limited with trade name Fructus Gardeniae product sold.The granularity of gardenia powder is generally 60 ~ 100 orders.
Petroleum hydrocarbon is also product sold in the market, such as by Guangzhou chemical reagents corporation with trade name petroleum ether product sold.
In this step, from gardenia powder, oil-soluble ingredients Fructus Gardeniae oil is extracted with petroleum hydrocarbon.
In this step, described filtering residue is when the 1st supersound extraction, and the weight ratio of gardenia powder and petroleum hydrocarbon is 90 ~ 110:750 ~ 850; When the 2nd time or the 3rd supersound extraction, the weight ratio of gardenia powder and petroleum hydrocarbon is 90 ~ 110:550 ~ 650.
The ultrasonic washing unit that this step uses and supersound process condition thereof as previously described, do not repeat them here.
C, prepare jasminoidin
800 ~ 1200 weight portions, 70 ~ 80% ethanol waters are by volume added in 80 ~ 120 weight portion gardenia powder, then in ultrasonic washing unit, supersound extraction is carried out, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 2 ~ 3 times in the same fashion, filtrate merges, then at flow velocity be under the condition of 20 ~ 30 microlitres/second by nonpolarity macroporous adsorptive resins chromatographic column and at flow velocity be 30 ~ 37 microlitres/second condition under pass through polyamide column, then working concentration 15 ~ 25% ethanol water eluting agent are by volume from adsorption column eluting, the eluent obtained processes in a rotary evaporator and obtains jasminoidin under the condition of distilling under reduced pressure,
The granularity of the gardenia powder that this step uses is generally 60 ~ 100 orders.
In this step, from gardenia powder, Alcohol soluble composition jasminoidin is extracted with ethanol water.
In this step, the ultrasonic washing unit of use and supersound process condition thereof as previously described, do not repeat them here.
In this step, the filtrate obtained is to be separated jasminoidin and Gardenia Yellow and other impurity by the object of nonpolar macroporous adsorption resin process.The macroporous adsorbent resin that the present invention uses is nonpolar D101, LX-60 or LX-20 type resin prepared by styrene and divinylbenzene combined polymerization, D101, LX-60, LX-20 macroporous adsorbent resin such as sold with trade name macroporous resin by Xi'an Sunresin New Materials Co., Ltd..
In the present invention, described filtrate is less than 20 microlitres/second by the flow velocity of described resin bed, then can affect the delivery time; If described flow velocity is greater than 30 microlitres/second, then can makes absorption and be separated not exclusively.Therefore, described flow velocity is 20 ~ 30 microlitres/second is appropriate, preferably 22 ~ 28 microlitres/second, more preferably 24 ~ 26 microlitres/second.
The effluent of described resin bed, then by polyamide column, its object is to be separated pigment and impurity component further, purification of samples.
Described polyamide synthesizes by caprolactam or adipic acid and hexamethylene diamine the high-molecular polyamide prepared.The high-molecular polyamide that the present invention uses is the polyamide sold with trade name polyamide by the biochemical plastic molding and processing plant of Taizhou City of Zhejiang Province road and bridge tetramethyl.
In the present invention, described resin bed effluent is less than 30 microlitres/second by the flow velocity of described polyamide column, then can affect the delivery time; If described flow velocity is greater than 37 microlitres/second, then separating effect can be affected.Therefore, described flow velocity is 30 ~ 37 microlitres/second is appropriate, preferably 32 ~ 36 microlitres/second, more preferably 34 ~ 36 microlitres/second.
Preferably, the eluant of described macroporous adsorptive resins or the eluant of described polyamide column are concentration 18 ~ 22% ethanol waters by volume.
Under the condition of pressure 0.002-0.098MPa, distilling under reduced pressure is carried out in the rotary evaporator that the eluent obtained is sold with trade name RE-301 type rotary evaporator in Guangzhou Xing Shuo Instrument Ltd..
D, prepare spirulina polysaccharide and spirulina polypeptide
Fresh spirulina is processed 12 ~ 18 minutes in low temperature biomixer under 4 DEG C ~ 6 DEG C cryogenic conditions, the treatment fluid obtained carries out centrifugal treating through low temperature supercentrifuge again under temperature 4 DEG C ~ 6 DEG C conditions, then filter at low temperature equipment is used to filter under the condition of temperature 2 DEG C ~ 5 DEG C, then the permeable membrane using molecular cut off to be 10kD under the condition of temperature 2 DEG C ~ 8 DEG C carries out membrane permeation process, obtains spirulina polysaccharide and spirulina polypeptide;
Spirulina (SpirulinaplatensisGeitl.) is a class unicellular lower eukaryote, prokaryote, the filamentous be made up of unicellular or many cells, the long 200-500 μm of body, wide 5-10 μm, cylindrical, in loose or closely well-regulated spiral rotary shape bend.It has the toxicity alleviating cancer radiation, chemotherapy, improves immunologic function, reduces the effects such as blood fat.
The low temperature biomixer that the present invention uses is product sold in the market, such as by company of Guangdong Juneng Biology & Technology Co., Ltd. with trade name JN-02C low temperature continuous flow cell disintegrating machine product sold.
In this step, the rotating speed of described low temperature supercentrifuge is 20000 ~ 30000 revs/min.The low temperature supercentrifuge that the present invention uses is product sold in the market, such as by company of Beckman Coulter Inc. of the U.S. with trade name XPN-80 supercentrifuge product sold.
Described filter at low temperature equipment is product sold in the market, such as by Dongguan Long Tian filter plant company limited with trade name LT600 insulation jacket filter product sold.
The permeable membrane that the present invention uses is product sold in the market, such as by Hydranautics with trade name QUALSEPDairyRO series concentrating and separating film product sold.
In this step, membrane permeation process carries out under 2 DEG C ~ 8 DEG C cryogenic conditions.
The product that membrane permeation process obtains adopts conventional infrared spectrum, its spirulina polysaccharide and spirulina polypeptide are determined in mass spectrometric analysis method analysis.
E, preparation chitosan spray
2.5 ~ 3.2 weight portions are dissolved in 6 ~ 15 weight portion concentration by weight in 0.8 ~ 1.2% aqueous acetic acid at the modification of chitosan that steps A obtains, then in obtained solution, 0.8 ~ 1.2 weight portion polysorbate60 is added, 0.8 ~ 1.2 weight portion lauroylamidopropyl betaine, mix homogeneously, add the jasminoidin that 0.3 ~ 0.8 weight portion obtains in step C again, the Fructus Gardeniae oil that 0.1 ~ 0.4 weight portion obtains in step B, the spirulina polypeptide that 0.1 ~ 0.4 weight portion obtains in step D, the spirulina polysaccharide that 0.1 ~ 0.4 weight portion obtains in step D, mix homogeneously, finally add 0.1 ~ 0.4 weight portion pi-allyl tributyl bromide phosphine, with distilled water, its gross weight is reached 100 weight portions.
Preferably, the weight ratio of modification of chitosan, polysorbate60, lauroylamidopropyl betaine, jasminoidin, Fructus Gardeniae oil, spirulina polypeptide, spirulina polysaccharide and pi-allyl tributyl bromide phosphine is 2.8 ~ 3.0:0.85 ~ 1.12:0.85 ~ 1.12:0.45 ~ 0.65:0.15 ~ 0.24:0.15 ~ 0.26:0.15 ~ 0.25:0.15 ~ 0.24.
The effect of the polysorbate60 that the present invention uses is to stablize Fructus Gardeniae oil and jasminoidin, makes it can not occur layering, segregation phenomenon in spray; It is product sold in the market, the polysorbate60 such as sold with trade name T-60 by Jiangsu Hai'an Petrochemical Plant.
The effect of lauroylamidopropyl betaine that the present invention uses is can dispersed spirulina polypeptide and spirulina polysaccharide, and make it there will not be precipitation, meanwhile, lauroylamidopropyl betaine also can play the effect promoting chitosan anti-bacteria usefulness further; It is product sold in the market, such as by upper marine Xiang Fine Chemical Co., Ltd with trade name CAB35 product sold.
The invention still further relates to the antibacterial modified chitosan spray adopting described preparation method to obtain.Antibacterial modified chitosan spray of the present invention detects through the antibacterial detection method of national standard (GB/T16886) and analyzes, and reaches more than 99% to the antibacterial ability of staphylococcus aureus, escherichia coli, streptococcus, Candida albicans, bacillus pyocyaneus or other fungus.
Antibacterial modified chitosan spray main body of the present invention adopts the natural products such as chitosan, jasminoidin, Fructus Gardeniae oil, spirulina polypeptide, spirulina polysaccharide, improves the biological safety of spray and the environmental protection of production process.
Spray of the present invention is by the collaborative impact of high deacetylized micromolecule chitosan and Fructus Gardeniae extract, spirulina polysaccharide extract, give full play to the antibacterial action of chitosan polysaccharide, greatly improve the overall anti-harmful microorganism effect of spray, to antibacterial, virus, fungus, all there is obvious killing effect.Meanwhile, the extract of Fructus Gardeniae and spirulina has biological activity, can reach the healing and growth that promote damaged skin, tissue, promote cellular metabolism, strengthen the resistance against diseases of skin.
The present invention is by adding a small amount of Fructus Gardeniae oil, polysorbate60 and lauroylamidopropyl betaine two kinds of non-ionic surface active agents, improve the diffusion performance of product, makes effective ingredient in spray more easily and epidermis generation effect, even diffusion, plays therapeutic effect.
Spray of the present invention is applied to treatment men and women's property pruritus vulvae and fungal infection, and courses of infection, folliculitis, vaginitis, is also applied to the treatment etc. of decubital ulcer and skin Chronic inflammation, applies very extensive.
[beneficial effect]
The invention has the beneficial effects as follows:
The mode that the present invention adopts two t-amyl peroxy compounds to combine with ultrasonic technique is with degrade chitosan; the inventive method adopts organic peroxide; action temperature and; reduce chitosan oxidation stain; again by molecule hyperfiltration technique; by chitosan classification; intercept the micromolecule chitosan of molecular weight lower than 500Da; then by deacetylated; acquisition low-molecular-weight is high deacetylized, the modification of chitosan of narrow molecular weight distribution; its free radical resisting and Scavenging ability improve greatly, and to improve anti-inflammatory activity, the ability to function of pathogenic microorganism also improves greatly.
Antibacterial modified chitosan spray main body of the present invention adopts the natural products such as chitosan, jasminoidin, Fructus Gardeniae oil, spirulina polypeptide, spirulina polysaccharide, improves the biological safety of spray and the environmental protection of production process.
Spray of the present invention is by the collaborative impact of high deacetylized micromolecule chitosan and Fructus Gardeniae extract, spirulina polysaccharide extract, give full play to the antibacterial action of chitosan polysaccharide, greatly improve the overall anti-harmful microorganism effect of spray, to antibacterial, virus, fungus, all there is obvious killing effect.Meanwhile, the extract of Fructus Gardeniae and spirulina has biological activity, can reach the healing and growth that promote damaged skin, tissue, promote cellular metabolism, strengthen the resistance against diseases of skin.
The present invention is by adding a small amount of Fructus Gardeniae oil, polysorbate60 and lauroylamidopropyl betaine two kinds of non-ionic surface active agents, improve the diffusion performance of product, makes effective ingredient in spray more easily and epidermis generation effect, even diffusion, plays therapeutic effect.
Spray of the present invention is applied to treatment men and women's property pruritus vulvae and fungal infection, and courses of infection, folliculitis, vaginitis, is also applied to the treatment etc. of decubital ulcer and skin Chronic inflammation, applies very extensive.
The antibacterial ability of spray of the present invention to staphylococcus aureus, escherichia coli, streptococcus, Candida albicans, bacillus pyocyaneus or other fungus reaches more than 99%.
[detailed description of the invention]
The present invention can be understood better by following embodiment.
Embodiment 1: prepare antibacterial modified chitosan spray
The implementation step of this embodiment is as follows:
A, prepare modification of chitosan
Sold with trade name food pharmaceutical grade chitosan by Jinan Haidebei Marine Organism Engineering Co., Ltd., deacetylation 80%, molecular weight 20 × 10
4the chitosan of Da, in 1.6% aqueous acetic acid by weight low whipping speed 250rpm condition under dissolve 5min, be incorporated as two t-amyl peroxy compounds of chitosan weight 1.0 times again, the solution obtained is in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited, supersound process 35min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W, obtain low-molecular weight chitoglycan complex solution, again through the polyacrylonitrile ultrafiltration film molecular retention sold with trade name UFbIA250-PAN film by Zhaoyuan Jinhuimo Technology Co., Ltd. and classification, obtain the chitosan that molecular weight is below 500Da, this chitosan is added to by weight in 35% potassium hydroxide aqueous solution, 10min is processed under the condition of temperature 105 DEG C, then with 1.6N aqueous hydrochloric acid solution carry out neutralizing, filter, distilled water wash, dry under the condition of temperature 60 C, obtain deacetylation 99%, molecular weight be the modification of chitosan of below 500Da,
B, prepare Fructus Gardeniae oil
700 weight portion petroleum hydrocarbons are added in 80 weight portion gardenia powder, then in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited under supersonic frequency 22kHz and the ultrasonic power condition that is 200W supersound process 35min, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 2 times in the same fashion with 500 weight portion petroleum ether, filtrate merges, employing heating in water bath mode removes the petroleum hydrocarbon in merging filtrate under the condition of temperature 60 C, obtains Fructus Gardeniae oil;
C, prepare jasminoidin
800 weight portions, 70% ethanol water is by volume added in 80 weight portion gardenia powder, then in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited, supersound process 35min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 3 times in the same fashion, filtrate merges, then be the D101 type macroporous adsorptive resins by being sold with trade name macroporous resin by Xi'an Sunresin New Materials Co., Ltd. under the condition of 26 microlitres/second at flow velocity, then the polyamide column by being sold with trade name polyamide by the biochemical plastic molding and processing plant of Taizhou City of Zhejiang Province road and bridge tetramethyl under the condition of flow velocity 30 microlitres/second, then working concentration 15% ethanol water eluting agent is by volume from adsorption column eluting, under the condition of pressure 0.002-0.098MPa, carry out distilling under reduced pressure in the rotary evaporator that the eluent obtained is sold with trade name RE-301 type rotary evaporator in Guangzhou Xing Shuo Instrument Ltd. and obtain jasminoidin,
D, prepare spirulina polysaccharide and spirulina polypeptide
Fresh spirulina is processed 12 minutes in the low temperature biomixer sold with trade name JN-02C low temperature continuous flow cell disintegrating machine by company of Guangdong Juneng Biology & Technology Co., Ltd. under temperature 4 DEG C of cryogenic conditions, the low temperature supercentrifuge of the treatment fluid obtained again through being sold with trade name XPN-80 supercentrifuge by company of Beckman Coulter Inc. of the U.S. carries out centrifugal treating under temperature 4 DEG C of conditions, then the filter at low temperature equipment sold with trade name LT600 insulation jacket filter by Dongguan Long Tian filter plant company limited is used to filter under the condition of temperature 2 DEG C, then use under the condition of temperature 2 DEG C and carry out membrane permeation process by Hydranautics with the permeable membrane that the molecular cut off that trade name QUALSEPDairyRO series concentrating and separating film is sold is 10kD, obtain spirulina polysaccharide and spirulina polypeptide,
E, preparation chitosan spray
2.5 weight portions are dissolved in 6 weight portion concentration by weight in 0.8% aqueous acetic acid at the modification of chitosan that steps A obtains, then in obtained solution, add the polysorbate60 that 0.8 weight portion is sold with trade name T-60 by Jiangsu Hai'an Petrochemical Plant, the lauroylamidopropyl betaine that 0.8 weight portion is sold with trade name CAB35 by upper marine Xiang Fine Chemical Co., Ltd, mix homogeneously, add the jasminoidin that 0.3 weight portion obtains in step C again, the Fructus Gardeniae oil that 0.1 weight portion obtains in step B, the spirulina polypeptide that 0.4 weight portion obtains in step D, the spirulina polysaccharide that 0.2 weight portion obtains in step D, mix homogeneously, finally add 0.1 weight portion pi-allyl tributyl bromide phosphine, with distilled water, its gross weight is reached 100 weight portions.
Determine according to the national standard antibacterial detection method detection analysis that this description describes, the antibacterial ability of antibacterial modified chitosan spray to staphylococcus aureus, escherichia coli, streptococcus, bacillus pyocyaneus prepared by the present embodiment all reaches 99.0%.
Embodiment 2: prepare antibacterial modified chitosan spray
The implementation step of this embodiment is as follows:
A, prepare modification of chitosan
Sold with trade name food pharmaceutical grade chitosan by Jinan Haidebei Marine Organism Engineering Co., Ltd., deacetylation 80%, molecular weight 24 × 10
4the chitosan of Da, in 3.0% aqueous acetic acid by weight low whipping speed 320rpm condition under dissolve 5min, be incorporated as two t-amyl peroxy compounds of chitosan weight 1.6 times again, the solution obtained is in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited, supersound process 38min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W, obtain low-molecular weight chitoglycan complex solution, again through the polyacrylonitrile ultrafiltration film molecular retention sold with trade name UFbIA250-PAN film by Zhaoyuan Jinhuimo Technology Co., Ltd. and classification, obtain the chitosan that molecular weight is below 500Da, this chitosan is added to by weight in 40% potassium hydroxide aqueous solution, 12min is processed under the condition of temperature 105 DEG C, then with 3.0N aqueous hydrochloric acid solution carry out neutralizing, filter, distilled water wash, dry under the condition of temperature 60 C, obtain deacetylation 100%, molecular weight be the modification of chitosan of below 500Da,
B, prepare Fructus Gardeniae oil
800 weight portion petroleum hydrocarbons are added in 110 weight portion gardenia powder, then in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited under supersonic frequency 22kHz and the ultrasonic power condition that is 200W supersound process 45min, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 3 times in the same fashion with 500 weight portion petroleum ether, filtrate merges, employing heating in water bath mode removes the petroleum hydrocarbon in merging filtrate under the condition of temperature 70 C, obtains Fructus Gardeniae oil;
C, prepare jasminoidin
1060 weight portions, 75% ethanol water is by volume added in 100 weight portion gardenia powder, then in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited, supersound process 45min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 2 times in the same fashion, filtrate merges, then be the LX-60 type resin macroporous adsorptive resins by being sold with trade name macroporous resin by Xi'an Sunresin New Materials Co., Ltd. under the condition of 20 microlitres/second at flow velocity, then the polyamide column by being sold with trade name polyamide by the biochemical plastic molding and processing plant of Taizhou City of Zhejiang Province road and bridge tetramethyl under the condition of flow velocity 32 microlitres/second, then working concentration 22% ethanol water eluting agent is by volume from adsorption column eluting, under the condition of pressure 0.002-0.098MPa, carry out distilling under reduced pressure in the rotary evaporator that the eluent obtained is sold with trade name RE-301 type rotary evaporator in Guangzhou Xing Shuo Instrument Ltd. and obtain jasminoidin,
D, prepare spirulina polysaccharide and spirulina polypeptide
Fresh spirulina is processed 15 minutes in the low temperature biomixer sold with trade name JN-02C low temperature continuous flow cell disintegrating machine by company of Guangdong Juneng Biology & Technology Co., Ltd. under 5 DEG C of cryogenic conditions, the low temperature supercentrifuge of the treatment fluid obtained again through being sold with trade name XPN-80 supercentrifuge by company of Beckman Coulter Inc. of the U.S. carries out centrifugal treating under temperature 4 DEG C of conditions, then the filter at low temperature equipment sold with trade name LT600 insulation jacket filter by Dongguan Long Tian filter plant company limited is used to filter under the condition of temperature 3 DEG C, then use under the condition of temperature 5 DEG C and carry out membrane permeation process by Hydranautics with the permeable membrane that the molecular cut off that trade name QUALSEPDairyRO series concentrating and separating film is sold is 10kD, obtain spirulina polysaccharide and spirulina polypeptide,
E, preparation chitosan spray
3.2 weight portions are dissolved in 14 weight portion concentration by weight in 1.0% aqueous acetic acid at the modification of chitosan that steps A obtains, then in obtained solution, add the polysorbate60 that 0.9 weight portion is sold with trade name T-60 by Jiangsu Hai'an Petrochemical Plant, the lauroylamidopropyl betaine that 0.9 weight portion is sold with trade name CAB35 by upper marine Xiang Fine Chemical Co., Ltd, mix homogeneously, add the jasminoidin that 0.5 weight portion obtains in step C again, the Fructus Gardeniae oil that 0.3 weight portion obtains in step B, the spirulina polypeptide that 0.2 weight portion obtains in step D, the spirulina polysaccharide that 0.4 weight portion obtains in step D, mix homogeneously, finally add 0.1 weight portion pi-allyl tributyl bromide phosphine, with distilled water, its gross weight is reached 100 weight portions.
Determine according to the national standard antibacterial detection method detection analysis that this description describes, the antibacterial ability of antibacterial modified chitosan spray to staphylococcus aureus, escherichia coli, streptococcus, bacillus pyocyaneus prepared by the present embodiment all reaches 99.6%.
Embodiment 3: prepare antibacterial modified chitosan spray
The implementation step of this embodiment is as follows:
A, prepare modification of chitosan
Sold with trade name food pharmaceutical grade chitosan by Jinan Haidebei Marine Organism Engineering Co., Ltd., deacetylation 80%, molecular weight 24 × 10
4the chitosan of Da, in 3.0% aqueous acetic acid by weight low whipping speed 350rpm condition under dissolve 10min, be incorporated as two t-amyl peroxy compounds of chitosan weight 2.0 times again, the solution obtained is in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited, supersound process 40min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W, obtain low-molecular weight chitoglycan complex solution, again through the polyacrylonitrile ultrafiltration film molecular retention sold with trade name UFbIA250-PAN film by Zhaoyuan Jinhuimo Technology Co., Ltd. and classification, obtain the chitosan that molecular weight is below 500Da, this chitosan is added to by weight in 38% potassium hydroxide aqueous solution, 18min is processed under the condition of temperature 110 DEG C, then with 1.0N aqueous hydrochloric acid solution carry out neutralizing, filter, distilled water wash, dry under the condition of temperature 60 C, obtain deacetylation 99%, molecular weight be the modification of chitosan of below 500Da,
B, prepare Fructus Gardeniae oil
840 weight portion petroleum hydrocarbons are added in 100 weight portion gardenia powder, then in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited under supersonic frequency 22kHz and the ultrasonic power condition that is 200W supersound process 42min, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 3 times in the same fashion with 700 weight portion petroleum ether, filtrate merges, employing heating in water bath mode removes the petroleum hydrocarbon in merging filtrate under the condition of temperature 60 C, obtains Fructus Gardeniae oil;
C, prepare jasminoidin
880 weight portions, 75% ethanol water is by volume added in 95 weight portion gardenia powder, then in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited, supersound process 45min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 2 times in the same fashion, filtrate merges, then be the LX-20 type resin macroporous adsorptive resins by being sold with trade name macroporous resin by Xi'an Sunresin New Materials Co., Ltd. under the condition of 24 microlitres/second at flow velocity, then the polyamide column by being sold with trade name polyamide by the biochemical plastic molding and processing plant of Taizhou City of Zhejiang Province road and bridge tetramethyl under the condition of flow velocity 34 microlitres/second, then working concentration 18% ethanol water eluting agent is by volume from adsorption column eluting, under the condition of pressure 0.002-0.098MPa, carry out distilling under reduced pressure in the rotary evaporator that the eluent obtained is sold with trade name RE-301 type rotary evaporator in Guangzhou Xing Shuo Instrument Ltd. and obtain jasminoidin,
D, prepare spirulina polysaccharide and spirulina polypeptide
Fresh spirulina is processed 18 minutes in the low temperature biomixer sold with trade name JN-02C low temperature continuous flow cell disintegrating machine by company of Guangdong Juneng Biology & Technology Co., Ltd. under 4 DEG C of cryogenic conditions, the low temperature supercentrifuge of the treatment fluid obtained again through being sold with trade name XPN-80 supercentrifuge by company of Beckman Coulter Inc. of the U.S. carries out centrifugal treating under temperature 6 DEG C of conditions, then the filter at low temperature equipment sold with trade name LT600 insulation jacket filter by Dongguan Long Tian filter plant company limited is used to filter under the condition of temperature 2 DEG C, then use under the condition of temperature 3 DEG C and carry out membrane permeation process by Hydranautics with the permeable membrane that the molecular cut off that trade name QUALSEPDairyRO series concentrating and separating film is sold is 10kD, obtain spirulina polysaccharide and spirulina polypeptide,
E, preparation chitosan spray
3.2 weight portions are dissolved in 14 weight portion concentration by weight in 1.0% aqueous acetic acid at the modification of chitosan that steps A obtains, then in obtained solution, add the polysorbate60 that 0.8 weight portion is sold with trade name T-60 by Jiangsu Hai'an Petrochemical Plant, the lauroylamidopropyl betaine that 0.9 weight portion is sold with trade name CAB35 by upper marine Xiang Fine Chemical Co., Ltd, mix homogeneously, add the jasminoidin that 0.5 weight portion obtains in step C again, the Fructus Gardeniae oil that 0.2 weight portion obtains in step B, the spirulina polypeptide that 0.4 weight portion obtains in step D, the spirulina polysaccharide that 0.1 weight portion obtains in step D, mix homogeneously, finally add 0.4 weight portion pi-allyl tributyl bromide phosphine, with distilled water, its gross weight is reached 100 weight portions.
Determine according to the national standard antibacterial detection method detection analysis that this description describes, the antibacterial ability of antibacterial modified chitosan spray to staphylococcus aureus, escherichia coli, streptococcus, bacillus pyocyaneus prepared by the present embodiment all reaches 99.8%.
Embodiment 4: prepare antibacterial modified chitosan spray
The implementation step of this embodiment is as follows:
A, prepare modification of chitosan
Sold with trade name food pharmaceutical grade chitosan by Jinan Haidebei Marine Organism Engineering Co., Ltd., deacetylation 80%, molecular weight 26 × 10
4the chitosan of Da, in 2.5% aqueous acetic acid by weight low whipping speed 250rpm condition under dissolve 8min, be incorporated as two t-amyl peroxy compounds of chitosan weight 2.0 times again, the solution obtained is in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited, supersound process 40min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W, obtain low-molecular weight chitoglycan complex solution, again through the polyacrylonitrile ultrafiltration film molecular retention sold with trade name UFbIA250-PAN film by Zhaoyuan Jinhuimo Technology Co., Ltd. and classification, obtain the chitosan that molecular weight is below 500Da, this chitosan is added to by weight in 42% potassium hydroxide aqueous solution, 10min is processed under the condition of temperature 115 DEG C, then with 1.0N aqueous hydrochloric acid solution carry out neutralizing, filter, distilled water wash, dry under the condition of temperature 60 C, obtain deacetylation 99%, molecular weight be the modification of chitosan of below 500Da,
B, prepare Fructus Gardeniae oil
860 weight portion petroleum hydrocarbons are added in 120 weight portion gardenia powder, then in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited under supersonic frequency 22kHz and the ultrasonic power condition that is 200W supersound process 42min, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 3 times in the same fashion with 700 weight portion petroleum ether, filtrate merges, employing heating in water bath mode removes the petroleum hydrocarbon in merging filtrate under the condition of temperature 60 C, obtains Fructus Gardeniae oil;
C, prepare jasminoidin
920 weight portions, 80% ethanol water is by volume added in 90 weight portion gardenia powder, then in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited, supersound process 42min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 3 times in the same fashion, filtrate merges, then be the D101 type resin macroporous adsorptive resins by being sold with trade name macroporous resin by Xi'an Sunresin New Materials Co., Ltd. under the condition of 30 microlitres/second at flow velocity, then the polyamide column by being sold with trade name polyamide by the biochemical plastic molding and processing plant of Taizhou City of Zhejiang Province road and bridge tetramethyl under the condition of flow velocity 36 microlitres/second, then working concentration 18% ethanol water eluting agent is by volume from adsorption column eluting, under the condition of pressure 0.002-0.098MPa, carry out distilling under reduced pressure in the rotary evaporator that the eluent obtained is sold with trade name RE-301 type rotary evaporator in Guangzhou Xing Shuo Instrument Ltd. and obtain jasminoidin,
D, prepare spirulina polysaccharide and spirulina polypeptide
Fresh spirulina is processed 18 minutes in the low temperature biomixer sold with trade name JN-02C low temperature continuous flow cell disintegrating machine by company of Guangdong Juneng Biology & Technology Co., Ltd. under 6 DEG C of cryogenic conditions, the low temperature supercentrifuge of the treatment fluid obtained again through being sold with trade name XPN-80 supercentrifuge by company of Beckman Coulter Inc. of the U.S. carries out centrifugal treating under temperature 5 DEG C of conditions, then the filter at low temperature equipment sold with trade name LT600 insulation jacket filter by Dongguan Long Tian filter plant company limited is used to filter under the condition of temperature 3 DEG C, then use under the condition of temperature 2 DEG C and carry out membrane permeation process by Hydranautics with the permeable membrane that the molecular cut off that trade name QUALSEPDairyRO series concentrating and separating film is sold is 10kD, obtain spirulina polysaccharide and spirulina polypeptide,
E, preparation chitosan spray
2.8 weight portions are dissolved in 10 weight portion concentration by weight in 1.1% aqueous acetic acid at the modification of chitosan that steps A obtains, then in obtained solution, add the polysorbate60 that 1.2 weight portions are sold with trade name T-60 by Jiangsu Hai'an Petrochemical Plant, the lauroylamidopropyl betaine that 1.0 weight portions are sold with trade name CAB35 by upper marine Xiang Fine Chemical Co., Ltd, mix homogeneously, add the jasminoidin that 0.8 weight portion obtains in step C again, the Fructus Gardeniae oil that 0.4 weight portion obtains in step B, the spirulina polypeptide that 0.2 weight portion obtains in step D, the spirulina polysaccharide that 0.3 weight portion obtains in step D, mix homogeneously, finally add 0.2 weight portion pi-allyl tributyl bromide phosphine, with distilled water, its gross weight is reached 100 weight portions.
Determine according to the national standard antibacterial detection method detection analysis that this description describes, the antibacterial ability of antibacterial modified chitosan spray to staphylococcus aureus, escherichia coli, streptococcus, bacillus pyocyaneus prepared by the present embodiment all reaches 99.1%.
Embodiment 5: prepare antibacterial modified chitosan spray
The implementation step of this embodiment is as follows:
A, prepare modification of chitosan
Sold with trade name food pharmaceutical grade chitosan by Jinan Haidebei Marine Organism Engineering Co., Ltd., deacetylation 80%, molecular weight 24 × 10
4the chitosan of Da, in 1.6% aqueous acetic acid by weight low whipping speed 270rpm condition under dissolve 10min, be incorporated as two t-amyl peroxy compounds of chitosan weight 2.5 times again, the solution obtained is in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited, supersound process 42min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W, obtain low-molecular weight chitoglycan complex solution, again through the polyacrylonitrile ultrafiltration film molecular retention sold with trade name UFbIA250-PAN film by Zhaoyuan Jinhuimo Technology Co., Ltd. and classification, obtain the chitosan that molecular weight is below 500Da, this chitosan is added to by weight in 45% potassium hydroxide aqueous solution, 12min is processed under the condition of temperature 105 DEG C, then with 3.0N aqueous hydrochloric acid solution carry out neutralizing, filter, distilled water wash, dry under the condition of temperature 60 C, obtain deacetylation 100%, molecular weight be the modification of chitosan of below 500Da,
B, prepare Fructus Gardeniae oil
820 weight portion petroleum hydrocarbons are added in 105 weight portion gardenia powder, then in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited under supersonic frequency 22kHz and the ultrasonic power condition that is 200W supersound process 45min, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 2 times in the same fashion with 500 weight portion petroleum ether, filtrate merges, employing heating in water bath mode removes the petroleum hydrocarbon in merging filtrate under the condition of temperature 70 C, obtains Fructus Gardeniae oil;
C, prepare jasminoidin
1000 weight portions, 75% ethanol water is by volume added in 100 weight portion gardenia powder, then in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited, supersound process 45min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 3 times in the same fashion, filtrate merges, then be the LX-60 type resin macroporous adsorptive resins by being sold with trade name macroporous resin by Xi'an Sunresin New Materials Co., Ltd. under the condition of 25 microlitres/second at flow velocity, then the polyamide column by being sold with trade name polyamide by the biochemical plastic molding and processing plant of Taizhou City of Zhejiang Province road and bridge tetramethyl under the condition of flow velocity 37 microlitres/second, then working concentration 20% ethanol water eluting agent is by volume from adsorption column eluting, under the condition of pressure 0.002-0.098MPa, carry out distilling under reduced pressure in the rotary evaporator that the eluent obtained is sold with trade name RE-301 type rotary evaporator in Guangzhou Xing Shuo Instrument Ltd. and obtain jasminoidin,
D, prepare spirulina polysaccharide and spirulina polypeptide
Fresh spirulina is processed 18 minutes in the low temperature biomixer sold with trade name JN-02C low temperature continuous flow cell disintegrating machine by company of Guangdong Juneng Biology & Technology Co., Ltd. under 6 DEG C of cryogenic conditions, the low temperature supercentrifuge of the treatment fluid obtained again through being sold with trade name XPN-80 supercentrifuge by company of Beckman Coulter Inc. of the U.S. carries out centrifugal treating under temperature 5 DEG C of conditions, then the filter at low temperature equipment sold with trade name LT600 insulation jacket filter by Dongguan Long Tian filter plant company limited is used to filter under the condition of temperature 3 DEG C, then use under the condition of temperature 2 DEG C and carry out membrane permeation process by Hydranautics with the permeable membrane that the molecular cut off that trade name QUALSEPDairyRO series concentrating and separating film is sold is 10kD, obtain spirulina polysaccharide and spirulina polypeptide,
E, preparation chitosan spray
2.6 weight portions are dissolved in 10 weight portion concentration by weight in 1.1% aqueous acetic acid at the modification of chitosan that steps A obtains, then in obtained solution, add the polysorbate60 that 1.0 weight portions are sold with trade name T-60 by Jiangsu Hai'an Petrochemical Plant, the lauroylamidopropyl betaine that 1.0 weight portions are sold with trade name CAB35 by upper marine Xiang Fine Chemical Co., Ltd, mix homogeneously, add the jasminoidin that 0.3 weight portion obtains in step C again, the Fructus Gardeniae oil that 0.4 weight portion obtains in step B, the spirulina polypeptide that 0.1 weight portion obtains in step D, the spirulina polysaccharide that 0.2 weight portion obtains in step D, mix homogeneously, finally add 0.3 weight portion pi-allyl tributyl bromide phosphine, with distilled water, its gross weight is reached 100 weight portions.
Determine according to the national standard antibacterial detection method detection analysis that this description describes, the antibacterial ability of antibacterial modified chitosan spray to staphylococcus aureus, escherichia coli, streptococcus, bacillus pyocyaneus prepared by the present embodiment all reaches 99.4%.
Embodiment 6: prepare antibacterial modified chitosan spray
The implementation step of this embodiment is as follows:
A, prepare modification of chitosan
Sold with trade name food pharmaceutical grade chitosan by Jinan Haidebei Marine Organism Engineering Co., Ltd., deacetylation 80%, molecular weight 24 × 10
4the chitosan of Da, in 1.6% aqueous acetic acid by weight low whipping speed 270rpm condition under dissolve 10min, be incorporated as two t-amyl peroxy compounds of chitosan weight 2.5 times again, the solution obtained is in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited, supersound process 42min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W, obtain low-molecular weight chitoglycan complex solution, again through the polyacrylonitrile ultrafiltration film molecular retention sold with trade name UFbIA250-PAN film by Zhaoyuan Jinhuimo Technology Co., Ltd. and classification, obtain the chitosan that molecular weight is below 500Da, this chitosan is added to by weight in 35% potassium hydroxide aqueous solution, 12min is processed under the condition of temperature 105 DEG C, then with 3.0N aqueous hydrochloric acid solution carry out neutralizing, filter, distilled water wash, dry under the condition of temperature 60 C, obtain deacetylation 99%, molecular weight be the modification of chitosan of below 500Da,
B, prepare Fructus Gardeniae oil
900 weight portion petroleum hydrocarbons are added in 90 weight portion gardenia powder, then in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited under supersonic frequency 22kHz and the ultrasonic power condition that is 200W supersound process 35min, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 2 times in the same fashion with 500 weight portion petroleum ether, filtrate merges, employing heating in water bath mode removes the petroleum hydrocarbon in merging filtrate under the condition of temperature 90 DEG C, obtains Fructus Gardeniae oil;
C, prepare jasminoidin
1200 weight portions, 74% ethanol water is by volume added in 120 weight portion gardenia powder, then in the ultrasonic washing unit sold with trade name YD1008 ultrasonic washing unit by Changzhou You Da ultrasonic equipment company limited, supersound process 38min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 2 times in the same fashion, filtrate merges, then be the LX-20 type resin macroporous adsorptive resins by being sold with trade name macroporous resin by Xi'an Sunresin New Materials Co., Ltd. under the condition of 23 microlitres/second at flow velocity, then the polyamide column by being sold with trade name polyamide by the biochemical plastic molding and processing plant of Taizhou City of Zhejiang Province road and bridge tetramethyl under the condition of flow velocity 35 microlitres/second, then working concentration 25% ethanol water eluting agent is by volume from adsorption column eluting, under the condition of pressure 0.002-0.098MPa, carry out distilling under reduced pressure in the rotary evaporator that the eluent obtained is sold with trade name RE-301 type rotary evaporator in Guangzhou Xing Shuo Instrument Ltd. and obtain jasminoidin,
D, prepare spirulina polysaccharide and spirulina polypeptide
Fresh spirulina is processed 14 minutes in the low temperature biomixer sold with trade name JN-02C low temperature continuous flow cell disintegrating machine by company of Guangdong Juneng Biology & Technology Co., Ltd. under 4 DEG C of cryogenic conditions, the low temperature supercentrifuge of the treatment fluid obtained again through being sold with trade name XPN-80 supercentrifuge by company of Beckman Coulter Inc. of the U.S. carries out centrifugal treating under temperature 6 DEG C of conditions, then the filter at low temperature equipment sold with trade name LT600 insulation jacket filter by Dongguan Long Tian filter plant company limited is used to filter under the condition of temperature 5 DEG C, then use under the condition of temperature 8 DEG C and carry out membrane permeation process by Hydranautics with the permeable membrane that the molecular cut off that trade name QUALSEPDairyRO series concentrating and separating film is sold is 10kD, obtain spirulina polysaccharide and spirulina polypeptide,
E, preparation chitosan spray
3.0 weight portions are dissolved in 8 weight portion concentration by weight in 1.0% aqueous acetic acid at the modification of chitosan that steps A obtains, then in obtained solution, add the polysorbate60 that 1.1 weight portions are sold with trade name T-60 by Jiangsu Hai'an Petrochemical Plant, the lauroylamidopropyl betaine that 1.2 weight portions are sold with trade name CAB35 by upper marine Xiang Fine Chemical Co., Ltd, mix homogeneously, add the jasminoidin that 0.4 weight portion obtains in step C again, the Fructus Gardeniae oil that 0.1 weight portion obtains in step B, the spirulina polypeptide that 0.4 weight portion obtains in step D, the spirulina polysaccharide that 0.3 weight portion obtains in step D, mix homogeneously, finally add 0.2 weight portion pi-allyl tributyl bromide phosphine, with distilled water, its gross weight is reached 100 weight portions.
Determine according to the national standard antibacterial detection method detection analysis that this description describes, the antibacterial ability of antibacterial modified chitosan spray to staphylococcus aureus, escherichia coli, streptococcus, bacillus pyocyaneus prepared by the present embodiment all reaches 99.6%.
Claims (10)
1. a preparation method for antibacterial modified chitosan spray, is characterized in that the step of this preparation method is as follows:
A, prepare modification of chitosan
By deacetylation 80%, molecular weight 20 × 10
4~ 30 × 10
4the chitosan of Da is dissolved in by weight in 1.0 ~ 3.0% aqueous acetic acids, be incorporated as two t-amyl peroxy compounds of chitosan weight 1 ~ 3 times again, the solution obtained carries out supersound process in ultrasonic washing unit, obtain low-molecular weight chitoglycan complex solution, again through polyacrylonitrile ultrafiltration film molecular retention and classification, obtain the chitosan that molecular weight is below 500Da; This chitosan is added to by weight in 35 ~ 45% potassium hydroxide aqueous solutions, in temperature 105 ~ 115
oprocess 10 ~ 20min under the condition of C, then use 1.0 ~ 3.0N aqueous hydrochloric acid solution to carry out neutralizing, filter, distilled water wash, drying, obtain deacetylation 99 ~ 100%, molecular weight be the modification of chitosan of below 500Da;
B, prepare Fructus Gardeniae oil
700 ~ 900 weight portion petroleum hydrocarbons are added in 80 ~ 120 weight portion gardenia powder, then in ultrasonic washing unit, supersound extraction is carried out, filtration obtains filtrate and filtering residue, described filtering residue then uses 500 ~ 700 weight portion petroleum hydrocarbons to repeat supersound extraction in the same fashion 2 ~ 3 times, filtrate merges, employing heating in water bath mode removes the petroleum hydrocarbon in merging filtrate under the condition of temperature 60 ~ 90 DEG C, obtains Fructus Gardeniae oil;
C, prepare jasminoidin
800 ~ 1200 weight portions, 70 ~ 80% ethanol waters are by volume added in 80 ~ 120 weight portion gardenia powder, then in ultrasonic washing unit, supersound extraction is carried out, filtration obtains filtrate and filtering residue, described filtering residue then repeats supersound extraction 2 ~ 3 times in the same fashion, filtrate merges, then be pass through polyamide column by nonpolarity macroporous adsorptive resins chromatographic column with under the condition of flow velocity 30 ~ 37 microlitres/second under the condition of 20 ~ 30 microlitres/second at flow velocity, then working concentration 15 ~ 25% ethanol water eluting agent are by volume from adsorption column eluting, the eluent obtained processes in a rotary evaporator and obtains jasminoidin under the condition of distilling under reduced pressure,
D, prepare spirulina polysaccharide and spirulina polypeptide
Fresh spirulina is processed 12 ~ 18 minutes in low temperature biomixer under 4 DEG C ~ 6 DEG C cryogenic conditions, the treatment fluid obtained carries out centrifugal treating through low temperature supercentrifuge again under temperature 4 DEG C ~ 6 DEG C conditions, then filter at low temperature equipment is used to filter under the condition of temperature 2 DEG C ~ 5 DEG C, then the permeable membrane using molecular cut off to be 10kD under the condition of temperature 2 DEG C ~ 8 DEG C carries out membrane permeation process, obtains spirulina polysaccharide and spirulina polypeptide;
E, preparation chitosan spray
2.5 ~ 3.2 weight portions are dissolved in 6 ~ 15 weight portion concentration by weight in 0.8 ~ 1.2% aqueous acetic acid at the modification of chitosan that steps A obtains, then in obtained solution, 0.8 ~ 1.2 weight portion polysorbate60 is added, 0.8 ~ 1.2 weight portion lauroylamidopropyl betaine, mix homogeneously, add the jasminoidin that 0.3 ~ 0.8 weight portion obtains in step C again, the Fructus Gardeniae oil that 0.1 ~ 0.4 weight portion obtains in step B, the spirulina polypeptide that 0.1 ~ 0.4 weight portion obtains in step D, the spirulina polysaccharide that 0.1 ~ 0.4 weight portion obtains in step D, mix homogeneously, finally add 0.1 ~ 0.4 weight portion pi-allyl tributyl bromide phosphine, with distilled water, its gross weight is reached 100 weight portions.
2. preparation method according to claim 1, is characterized in that in step, dissolves 5 ~ 15min under the condition of deacetylation 80% chitosan low whipping speed 250 ~ 350rpm in aqueous acetic acid.
3. preparation method according to claim 1, is characterized in that in step, and the solution obtained is supersound process 35 ~ 45min under supersonic frequency 22kHz and ultrasonic power are the condition of 200W.
4. preparation method according to claim 1, is characterized in that in stepb, and described filtering residue is when the 1st supersound extraction, and the weight ratio of gardenia powder and petroleum hydrocarbon is 90 ~ 110:750 ~ 850; When the 2nd time or the 3rd supersound extraction, the weight ratio of gardenia powder and petroleum hydrocarbon is 90 ~ 110:550 ~ 650.
5. preparation method according to claim 1, is characterized in that in step C, and described macroporous adsorbent resin is nonpolar D101, LX-60 or LX-20 type resin prepared by styrene and divinylbenzene combined polymerization; Described polyamide synthesizes by caprolactam or adipic acid and hexamethylene diamine the high-molecular polyamide prepared.
6. preparation method according to claim 1, is characterized in that in step C, and the eluant of described macroporous adsorptive resins or the eluant of described polyamide column are concentration 18 ~ 22% ethanol waters by volume.
7. preparation method according to claim 1, is characterized in that in step D, and described permeable membrane is the permeable membrane prepared by polyether sulfone, Triafol T or regenerated cellulose.
8. preparation method according to claim 1, is characterized in that in step D, and the rotating speed of described low temperature supercentrifuge is 20000 ~ 30000 revs/min.
9. preparation method according to claim 1, is characterized in that the weight ratio of modification of chitosan, polysorbate60, lauroylamidopropyl betaine, jasminoidin, Fructus Gardeniae oil, spirulina polypeptide, spirulina polysaccharide and pi-allyl tributyl bromide phosphine is 2.8 ~ 3.0:0.85 ~ 1.12:0.85 ~ 1.12:0.45 ~ 0.65:0.15 ~ 0.24:0.15 ~ 0.26:0.15 ~ 0.25:0.15 ~ 0.24.
10. the antibacterial modified chitosan spray that preparation method described in claim obtains any one of claim 1-9, is characterized in that it reaches more than 99% to the antibacterial ability of staphylococcus aureus, escherichia coli, streptococcus, Candida albicans or bacillus pyocyaneus.
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| CN105418779A (en) * | 2015-09-10 | 2016-03-23 | 周玥 | Spirulina polysaccharide extraction process and applications thereof |
| CN105670335A (en) * | 2015-12-31 | 2016-06-15 | 江门市林业科学研究所 | Method for two-pot two stage countercurrent dynamic extraction of gardenia yellow pigment |
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