CN109303930A - A kind of Medical absorbable antiseptic dressing and preparation method thereof - Google Patents
A kind of Medical absorbable antiseptic dressing and preparation method thereof Download PDFInfo
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- CN109303930A CN109303930A CN201811133117.1A CN201811133117A CN109303930A CN 109303930 A CN109303930 A CN 109303930A CN 201811133117 A CN201811133117 A CN 201811133117A CN 109303930 A CN109303930 A CN 109303930A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/225—Mixtures of macromolecular compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
- A61L15/46—Deodorants or malodour counteractants, e.g. to inhibit the formation of ammonia or bacteria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
- A61L15/62—Compostable, hydrosoluble or hydrodegradable materials
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/04—Materials for stopping bleeding
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Abstract
The present invention relates to a kind of Medical absorbable antiseptic dressing and preparation method thereof, include the following steps: rat-tail tendon obtaining collagen supernatant by pretreatment, sour enzyme combination processing, by collagen supernatant through oversalting, dialysis, freeze-drying；It dissolves the chitosan in acid, process and acetic anhydride, obtains N- Acetylated Chitosans precipitating titrating by titer, N- Acetylated Chitosans precipitating is filtered, is dry；Collagen and N- Acetylated Chitosans are mixed in 0.05-0.5mol/L acid and form mixed liquor; wherein; the mass volume ratio of collagen and N- acetylated chitosan sugar juice in mixed liquor is 0.5%-5%; the mass ratio of collagen and N- Acetylated Chitosans is (6-9): (1-4), by the dry acquisition Medical absorbable antiseptic dressing of mixed liquor.The two is combined and makes it have synergistic effect by the present invention, and obtained dressing not only has good biocompatibility and degradability, but also plays the role of hemostasis, analgesic and antibacterial early period in wound healing.
Description
Technical field
The present invention relates to Iatrochemistry Material Fields, more particularly, to a kind of Medical absorbable antiseptic dressing and its system
Preparation Method.
Background technique
Collagen is a kind of high molecular weight protein generally existing in animal body, be extracellular matrix important composition at
Point, it is primarily present in the connective tissue of animal, accounts for the 25%~30% of mammalian body internal protein.Collagen is because having
Good biocompatibility, biodegradable, low antigenicity, at the same again can active cell characteristic gene expression, maintain thin
The normal characteristic expression of born of the same parents, is conducive to adherency, growth, proliferation and the differentiation of cell, thus is largely used to bio-medical material
Field.The type of collagen is more, and common type is I type, II type, III type, and wherein Type I collagen albumen is distributed mainly on skin
In the tissue such as skin, tendon, bone, it accounts for the 90% of organism whole collagen content, is that field of tissue engineering technology is ground in recent years
The hot spot studied carefully.
Chitosan is the deacetylated product of natural polysaccharide chitosan moiety, is unique positively charged alkaline polysaccharide in nature.
The structural units of chitosan molecule chain is amino and acetylglucosamine, has good cell affinity and bio-compatible
Property, while there are the different physiological roles such as hemostasis, antibacterial, wound healing promoting, obtain it widely in biomedicine field
Using.
Currently, most of medical materials in the market only play the role of package, mechanical property and degradation property compared with
Difference, and anti-microbial property and tissue repair performance is ineffective.
Summary of the invention
The present invention provides a kind of Medical absorbable antiseptic dressing and preparation method thereof, which has preferable mechanical property
Energy and degradation property, while but also with good antibiotic property and tissue repair performance.
To achieve the above object, the present invention adopts the following technical scheme:
A kind of preparation method of Medical absorbable antiseptic dressing, includes the following steps:
Rat-tail tendon is obtained into collagen supernatant by pretreatment, sour enzyme combination processing, collagen supernatant is passed through
Oversalting, dialysis, freeze-drying, can be obtained collagen；
It dissolves the chitosan in acid, process and acetic anhydride, obtains N- acetylated chitosan titrating by titer
N- Acetylated Chitosans can be obtained in the filtering of N- Acetylated Chitosans precipitating, drying by sugar precipitating；
The collagen and N- Acetylated Chitosans are mixed in 0.05-0.5mol/L acid and form mixed liquor, wherein
The mass volume ratio of collagen and N- acetylated chitosan sugar juice in the mixed liquor is 0.5%-5%, the collagen
The mass ratio of albumen and N- Acetylated Chitosans is (6-9): (1-4), and the mixed liquor is dry, and it is anti-to obtain Medical absorbable
Bacterium dressing.
The pre-treatment step includes: and cleans by rat-tail tendon after grease removal, inactivation treatment in one of the embodiments,
It smashs to pieces, it is dry.
The sour enzyme combination processing step includes: to immerse pretreated rat-tail tendon in one of the embodiments,
1-6h is dissolved in the acetic acid of 0.05-0.5mol/L, 1-4 parts of pepsins are being added, 24-72h is persistently stirred and obtains collagen
The collagen extracting solution refrigerated centrifuge is obtained collagen supernatant by extracting solution.
The revolving speed of the refrigerated centrifuge is 3000-12000rad/min in one of the embodiments, and centrifugation time is
10-30min, the temperature of freezing are 4 DEG C.
The salting-out step includes: to add hydroxide into the collagen supernatant in one of the embodiments,
Sodium solution is adjusted to neutrality, is 2-3mol/L salt is added to adjust the concentration of collagen supernatant, 12-24h is stirred, to collagen egg
White Precipitation.
The dialysis step includes: that the collagen deposit is dissolved in 0.05- in one of the embodiments,
It is outer dialyzate dialysis 2d with the acetic acid of 0.05-0.5mol/L, replacement daily is twice in the acetic acid of 0.5mol/L；Again with distillation
Water is outer dialyzate dialysis 2d, daily replacement 2 times.
The mass ratio of the chitosan and the acetic anhydride is (1-4) in one of the embodiments: 4.
The titer is the ethanol-water solution of potassium hydroxide in one of the embodiments, wherein the hydroxide
The concentration of potassium is 0.5-1mol/L, and the volume ratio of ethyl alcohol and water is 1:1.
The mixed liquor freezes 1-2h in -80 DEG C in one of the embodiments, is freeze-dried 24-48h.
The present invention also provides Medical absorbable antiseptic dressings made from a kind of preparation method.
Collagen can be used for wound dressing, hemostasis, while also have the advantages that degradation time is short；N- acetylated chitosan
Sugar has good cellular affinity and biocompatibility, also has the advantages that hemostasis, antibacterial, promotion wound healing.The present invention
The two is combined and makes it have synergistic effect, obtained dressing not only has good biocompatibility, but also is cured in wound
It closes and plays the role of hemostasis, analgesic and antibacterial early period.In addition, the dressing degradation time is short, the residual such as cross-linking agent-free, therefore, energy
Enough adhesives that dressing and tissue are reduced in wound healing mid-term.
Detailed description of the invention
Fig. 1 is the degradation figure of collagen.
Specific embodiment
In order to make the foregoing objectives, features and advantages of the present invention clearer and more comprehensible, with reference to the accompanying drawing to the present invention
Specific embodiment be described in detail.Many details are explained in the following description in order to fully understand this hair
It is bright.But the invention can be embodied in many other ways as described herein, those skilled in the art can be not
Similar improvement is done in the case where violating intension of the present invention, therefore the present invention is not limited to the specific embodiments disclosed below.
Unless otherwise defined, all technical and scientific terms used herein and belong to technical field of the invention
The normally understood meaning of technical staff is identical.Term as used herein in the specification of the present invention is intended merely to description tool
The purpose of the embodiment of body, it is not intended that in the limitation present invention.
Embodiment 1
1) by coypu tail tendon after grease removal, inactivation treatment, clean cutting, which is put into tissue mashing machine, is smashed to pieces, and cleaning is smashed to pieces
Tail tendon after, spontaneously dry.
2) 20 parts of tail tendons are immersed in 0.5mol/L acetum, are stirred to dissolve.Adding 1 part of pepsin supreme
It states in solution, persistently stirs 72h, obtain collagen extracting solution.Collagen extracting solution is subjected to refrigerated centrifuge, centrifuge temperature
Degree is 4 DEG C, revolving speed 12000r/min, centrifugation time 10min, obtains collagen supernatant.
3) the 10mol/LNaOH solution of the collagen supernatant after centrifugation is adjusted into pH to neutrality, NaCl is added, makes it
Concentration reaches 2mol/L, stirs 12h, collagen deposit is precipitated, and filtering obtains collagen solids.
4) 0.5mol/L acetum is added into collagen solids and dissolves collagen solids.With 0.5mol/L acetic acid
Solution be extracellular fluid dialysis dialyse 2d, daily replacement 2 times, then with distilled water be outer dialyzate dialyse 2d, daily replacement 2 times.By glue
Former protein frozen is dry, saves backup.
5) 1 part of chitosan is dissolved in 20 parts of 0.5mol/L acetums, adds 20 parts of methanol solution dilution chitosans, stirs
It mixes and mixes 10min.
6) acetic anhydride is added into the solution of step 5), makes the mass ratio 1:4 of acetic anhydride and chitosan, stir 15min,
Stand 2h.
7) precipitating, mistake is precipitated in the mixed liquor of step 6) alcohol-water (V:V=1:1) solution of the KOH of 0.5mol/L
Filter.It is washed with distilled water to neutrality, it is dry in 60 DEG C of vacuum tanks.N- Acetylated Chitosans are crushed, are stored after dry.
8) collagen and N- Acetylated Chitosans are added separately in 0.5mol/L acetum, collagen is molten
The mixed liquor for being configured to mass volume ratio and being 5% of liquid and N- acetylated chitosan sugar juice.Wherein, collagen solution and N- second
The mass ratio 6:4 of acylation chitosan solution.
9) mixed liquor of step 8) is freezed at -80 DEG C 1h, is placed in freeze drier and is freeze-dried 48h, cured
With absorbable antiseptic dressing.
Embodiment 2
1) by coypu tail tendon after grease removal, inactivation treatment, clean cutting, which is put into tissue mashing machine, is smashed to pieces, and cleaning is smashed to pieces
Tail tendon after, spontaneously dry.
2) 20 parts of tail tendons are immersed in 0.1mol/L acetum, are stirred to dissolve.Adding 2 parts of pepsins supreme
It states in mixed liquor, persistently stirs 48h, obtain collagen extracting solution.Collagen extracting solution is subjected to refrigerated centrifuge, centrifuge
Temperature is 4 DEG C, revolving speed 8000r/min, centrifugation time 20min and takes collagen supernatant.
3) the 10mol/L NaOH solution of the collagen supernatant after centrifugation is adjusted into pH to neutrality, NaCl is added, makes
Its concentration reaches 2.5mol/L, and stirring for 24 hours, is precipitated collagen deposit, and filtering obtains collagen solids.
4) 0.1mol/L acetum is added into collagen solids and dissolves collagen solids.With 0.1mol/L acetic acid
Solution be extracellular fluid dialysis dialyse 2d, daily replacement 2 times, then with distilled water be outer dialyzate dialyse 2d, daily replacement 2 times.By glue
Former protein frozen is dry, saves backup.
5) 1 part of chitosan is dissolved in 40 parts of 0.1mol/L acetums, adds 40 parts of methanol solution dilution chitosans, stirs
It mixes and mixes 10min.
6) acetic anhydride is added into the solution of step 5), makes the mass ratio 2:4 of acetic anhydride and chitosan, stir 15min,
Stand 3h.
7) precipitating is precipitated in the mixed liquor of step 6) alcohol-water (V:V=1:1) solution of the KOH of 1mol/L, filtered,
It is washed with distilled water to neutrality, it is dry in 60 DEG C of vacuum tanks.N- Acetylated Chitosans are crushed, are stored after dry.
8) collagen and N- Acetylated Chitosans are added separately in 0.1mol/L acetum, collagen is molten
The mixed liquor for being configured to mass volume ratio and being 2% of liquid and N- acetylated chitosan sugar juice.Wherein, collagen solution and N- second
Acylation chitosan solution 7:3 in mass ratio is uniformly mixed.
9) collagen and N- Acetylated Chitosans mixed liquor are freezed at -80 DEG C 2h, are placed in cold in freeze drier
Dry 48h is lyophilized, obtains Medical absorbable antiseptic dressing.
Embodiment 3
1) by coypu tail tendon after grease removal, inactivation treatment, clean cutting, which is put into tissue mashing machine, is smashed to pieces, and cleaning is smashed to pieces
Tail tendon after, spontaneously dry.
2) 20 parts of tail tendons are immersed in 0.05mol/L acetum, are stirred to dissolve.Plus 4 parts of pepsins then
Into mixed liquor, 72h is persistently stirred, obtains collagen extracting solution.Collagen extracting solution is subjected to refrigerated centrifuge, centrifuge
Temperature is 4 DEG C, revolving speed 10000r/min, centrifugation time 10min and takes collagen supernatant.
3) the 10mol/L NaOH solution of the collagen supernatant after centrifugation is adjusted into pH to neutrality, NaCl is added, makes
Its concentration reaches 3mol/L, and stirring for 24 hours, is precipitated collagen deposit, and filtering obtains collagen solids.
4) 0.05mol/L acetum is added into collagen solids and dissolves collagen solids.With 0.05mol/L vinegar
Acid solution be extracellular fluid dialysis dialyse 2d, daily replacement 2 times, then with distilled water be outer dialyzate dialyse 2d, daily replacement 2 times.It will
Collagen freeze-drying, saves backup.
5) 1 part of chitosan is dissolved in 60 parts of 0.05mol/L acetums, adds 60 parts of methanol solution dilution chitosans,
Stir and evenly mix 10min.
6) acetic anhydride is added into the solution in step 5), makes the mass ratio 4:4 of acetic anhydride and chitosan, stir
15min stands 2h.
7) precipitating is precipitated in the mixed liquor of step 6) alcohol-water (V:V=1:1) solution of the KOH of 0.5~1mol/L,
Filtering, is washed with distilled water to neutrality, dry in 60 DEG C of vacuum tanks.N- Acetylated Chitosans are crushed, are stored after dry.
8) collagen and N- Acetylated Chitosans are added separately in 0.05mol/L acetum, by collagen
The mixed liquor for being configured to mass volume ratio and being 0.5% of solution and N- acetylated chitosan sugar juice.Wherein, collagen solution
It is uniformly mixed with N- acetylated chitosan sugar juice 8:2 in mass ratio.
9) collagen and N- Acetylated Chitosans mixed liquor are placed in freeze drier and are freezed in -80 DEG C of freezing 1h
Dry 48h, obtains Medical absorbable antiseptic dressing.
Embodiment 4
The mass volume ratio of collagen solution and N- Acetylated Chitosans is configured as 5% mixed liquor, is freeze-dried.It will
The dressing of above-mentioned drying is placed in 0.01mg/L clostridiopetidase A and 0.05g/L bacteriolyze enzyme buffer liquid.Clostridiopetidase A and bacteriolyze enzyme buffer liquid
Replacement is primary weekly.Degradation sample is periodically taken out, weighs after dry and calculates degradation rate.
Note: I represents the degradation curve of pure collagen, and II to represent collagen and N- Acetylated Chitosans mass ratio be 8:
2, III, which represents collagen and N- Acetylated Chitosans mass ratio, represents collagen and N- acetylated chitosan saccharic as 7:3, IV
For amount than being 6:4, V represents pure N- Acetylated Chitosans.
From attached drawing 1 it is found that collagen faster degradation is occurred by the effect of clostridiopetidase A, and N- Acetylated Chitosans
Can also degrade by the effect of lysozyme, collagen and N- Acetylated Chitosans it is compound after maintain the drops of two kinds of materials
Solution, the compositely proportional of collagen is higher, and the degradation rate of the dressing of preparation within a certain period of time is bigger.
Embodiment 5
The mass volume ratio of collagen solution and N- Acetylated Chitosans is configured as 5% mixed liquor, is freeze-dried.It will
Above-mentioned sample is added in 0.1g 0.5mol/L acetum, the above-mentioned solution of 5mL is measured and is added to 15ml fluid nutrient medium, adopt
Adjust pH to 6.0 with the NaOH solution of 0.1mol/L, and make the concentration of N- Acetylated Chitosans be respectively 0,0.1%,
0.15%, 2%, 5%.Culture medium is sub-packed in 5 test tubes, every pipe 8ml, 121 ° of sterilizing 15min, with sterile shifting after cooling
Liquid pipe is adsorbed bacteria suspension 0.3ml and is added in test tube, and slight oscillatory is uniform；It is measured under 610nm wavelength with spectrophotometer initial
Then absorbance value is placed on culture in 37 DEG C of constant incubators, measures absorbance value, afterwards for 24 hours with absorbance value reacting bacteria number
The variation of amount.Using the sample without N- acetyl chitosan as control experiment group, every group is not added sample as control, other conditions
It is identical.
Bacteriostasis rate=(control bacterium solution absorbance-N- Acetylated Chitosans bacterium solution absorbance)/control bacterium solution absorbance ×
100%
| C% (N- Acetylated Chitosans) | 0 | 0.1% | 0.15% | 2% | 5% |
| Bacteriostasis rate | 0 | 72.2% | 78.5% | 82.1% | 90.6% |
Above-mentioned test result explanation, N- Acetylated Chitosans have good anti-microbial property, the N- acetylation of various concentration
Chitosan shows different bacteriostasis properties, therefore, the dressing that anti-microbial property can be selected different according to the gradient of infection of wound.
Each technical characteristic of embodiment described above can be combined arbitrarily, for simplicity of description, not to above-mentioned reality
It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, all should be considered as described in this specification.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that coming for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to protection of the invention
Range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (10)
1. a kind of preparation method of Medical absorbable antiseptic dressing, which comprises the steps of:
Rat-tail tendon is obtained into collagen supernatant by pretreatment, sour enzyme combination processing, by collagen supernatant through supersalt
Analysis, dialysis, drying, can be obtained collagen；
It dissolves the chitosan in acid, process and acetic anhydride, it is heavy titrating acquisition N- Acetylated Chitosans by titer
It forms sediment, by the filtering of N- Acetylated Chitosans precipitating, drying, N- Acetylated Chitosans can be obtained；
The collagen and N- Acetylated Chitosans are mixed in 0.05-0.5mol/L acid and form mixed liquor, wherein is described
The mass volume ratio of collagen and N- acetylated chitosan sugar juice in mixed liquor is 0.5%-5%, the collagen
Mass ratio with N- Acetylated Chitosans is (6-9): (1-4), and the mixed liquor is dry, obtains Medical absorbable antibacterial and applies
Material.
2. preparation method according to claim 1, which is characterized in that the pre-treatment step includes: by rat-tail tendon through removing
After rouge, inactivation treatment, cleans and smash to pieces, it is dry.
3. preparation method according to claim 1, which is characterized in that the acid enzyme combination processing step includes: by pre- place
1-6h is dissolved in the acetic acid of rat-tail tendon immersion 0.05-0.5mol/L after reason, 1-4 parts of pepsins are being added, are persistently stirring 24-
72h obtains extracting solution, and the extracting solution refrigerated centrifuge is obtained collagen supernatant.
4. preparation method according to claim 3, which is characterized in that the revolving speed of the refrigerated centrifuge is 3000-
12000rad/min, centrifugation time 10-30min, the temperature of freezing are 4 DEG C.
5. preparation method according to claim 1, which is characterized in that the salting-out step includes: to the collagen
Sodium hydroxide solution is added in supernatant and is adjusted to neutrality, is 2-3mol/L salt is added to adjust concentration, 12-24h is stirred, to collagen
Albumen precipitation is precipitated.
6. preparation method according to claim 5, which is characterized in that the dialysis step includes: by the collagen
Precipitating is dissolved in the acetic acid of 0.05-0.5mol/L, is outer dialyzate dialysis 2d with the acetic acid of 0.05-0.5mol/L, is replaced daily
Twice；It is again outer dialyzate dialysis 2d, daily replacement 2 times with distilled water.
7. preparation method according to claim 1, which is characterized in that the mass ratio of the chitosan and the acetic anhydride is
(1-4): 4.
8. preparation method according to claim 1, which is characterized in that the titer is that the alcohol-water of potassium hydroxide is molten
Liquid, wherein the concentration of the potassium hydroxide solution is 0.5-1mol/L, and the volume ratio of ethyl alcohol and water is 1:1.
9. preparation method according to claim 1, which is characterized in that the mixed liquor freezes 1-2h in -80 DEG C, freezing
Dry 24-48h.
10. according to right to go any one of 1 to 10 preparation method obtained by Medical absorbable antiseptic dressing.
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