CN104491913A - Method for preparing silkworm chitosan composite haemostatic material - Google Patents
Method for preparing silkworm chitosan composite haemostatic material Download PDFInfo
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- CN104491913A CN104491913A CN201410773223.1A CN201410773223A CN104491913A CN 104491913 A CN104491913 A CN 104491913A CN 201410773223 A CN201410773223 A CN 201410773223A CN 104491913 A CN104491913 A CN 104491913A
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Abstract
The invention relates to a method for preparing a silkworm chitosan composite haemostatic material. The method comprises the following steps: (1) adding silkworm chitosan powder into 2 percent of glacial acetic acid, stirring until the powder is completely dissolved to form 1.0-2.5 mass percent of silkworm chitosan solution, mixing 3.0 mass percent of gelatin into the solution at the constant temperature of 60 DEG C, and stirring until the material is completely dissolved; (2) cooling to room temperature, sequentially dripping 0.005-0.020 mass percent of glutaraldehyde solution, 0.1-0.4 mass percent of calcium chloride solution and 0.05-0.2 mass percent of glycerin, and uniformly stirring and mixing; and (3) stirring and foaming the prepared blended solution at room temperature for 30 minutes by using a magnetic stirrer, pouring the solution into a culture dish paved with a polyethylene film, pre-freezing in an ultra-low temperature refrigerator at the pre-freezing temperature of 40 DEG C below zero for 1 hour, and finally drying in a lyophilization freeze drying machine for 30 hours, thereby obtaining the silkworm chitosan composite haemostatic material.
Description
Technical field
The invention belongs to Chinese patent medicine particles field, especially a kind of preparation method of silkworm chitosan compound hemostatic material.
Background technology
Stanch fibre is a kind of medical fiber material, because fibrous material surface amasss more much bigger than the surface area of general medical film, sponge etc., the contact area of itself and hemorrhage wound surface blood can be expanded, platelet ratio is made to be easier to adhere to or adsorb on the fiber surface, thus be conducive to solidifying of hemorrhage wound surface blood, reach object of stopping blooding rapidly.According to the retrieval, current domestic widely used hemostatic material has respective weak point, and the efficient hemostatic material of external import is because of its widespread adoption in daily hemostasis of price limit of its costliness.Existing stanch fibre is main component manufacture mainly with gelatin, and process is also different, and product quality is uneven, and real world applications is unsatisfactory.
Summary of the invention
The object of the invention is to overcome prior art weak point, the preparation method of the good silkworm chitosan compound hemostatic material of a kind of wound healing promoting performance, bacteriostasis property is provided.
Technical scheme of the present invention is:
A preparation method for silkworm chitosan compound hemostatic material, the steps include:
(1) silkworm chitosan powder is put into the glacial acetic acid of 2%, is stirred to and dissolves completely, form the silkworm chitosan solution that mass fraction is 1.0% ~ 2.5%, then at constant temperature 60 DEG C by mass fraction be 3.0% gelatin be mixed into wherein to be stirred to and dissolve completely;
(2) after being cooled to room temperature, drip successively mass fraction be 0.005% ~ 0.020% glutaraldehyde solution, mass fraction be 0.1% ~ 0.4% calcium chloride solution, mass fraction be the glycerol of 0.05% ~ 0.2%, be uniformly mixed;
(3) the above-mentioned blend solution magnetic stirring apparatus prepared at room temperature is carried out the stirring foaming of 30min, then pour into and be covered with in the culture dish of polyethylene film, schedule to last at pre-freezing temperature in the ultra cold storage freezer of-40 DEG C and carry out 1h pre-freeze, finally put into the dry 30h of frozen vacuum dryer, i.e. obtained silkworm chitosan compound hemostatic material.
And, after described silkworm chitosan compound hemostatic material ultra violet lamp sterilizing 20min, to preserve under Polythene Bag encapsulation room temperature.
And, by the preparation method of silkworm chitosan powder be: Pupa bombycis shell through de-inorganic salt, alkali deproteinization, washing and drying, decolouring drying, chitin, de-second phthalein, washing filtering and drying, obtains chitosan, gets product after crushed successively.
And described de-inorganic salt adopts the HCl of 3%, and soaking temperature is 30 DEG C, and soak time is 18h.
And during described alkali deproteinization, adopt the NaOH solution of 5% to embathe deproteinising, embathing temperature is 85 DEG C, and the time of embathing is 4h.
And, when described decolouring is dry, adopt mass fraction to be the H of 20%
2o
2solution, pH=11, treatment temperature are 80 DEG C, and the reason time is 2h, and bath raio is 1:40.
Advantage of the present invention and beneficial effect are:
1. staltic cellulose fibre raw material Cicadae puparium polysaccharide wide material sources
State of Shi Can sparetime university of China, the Bombyx bombycis great majority that people obtain by breeding silkworms for a long time are in order to reel silk from cocoons raw silk, and one of by-product of filature Pupa bombycis is taken as waste material abandons and it is reported, usual 1.5 ten thousand tons of Bombyx bombycis can produce 1 kiloton raw silk, and often produce the dry Pupa bombycis that 1 kiloton raw silk can obtain about 1 kiloton, the annual Main By product Pupa bombycis of China's filature industry can reach 530,00.More than ton, as can be seen here, the output of Pupa bombycis is considerable.
Although the chitin of extensive use at present and chitosan great majority extract from Crusta Penaeus seu Panulirus Carapax Eriocheir sinensis, but be rich in pupa oil and protein in Pupa bombycis, pupa slag is then that Pupa bombycis is through extracting the garbage after pupa oil and protein, wherein containing a large amount of unemployed chitin, and due to the content of ashes in Pupa bombycis less, so be chitin extraction, so prepare chitosan good raw material we can see, the content of chitosan in Pupa bombycis, up to 33.7-44.2%, is appreciable.
Development and utilization Pupa bombycis resource, both can turn waste into wealth, improve Business Economic Benefit, the shortage problem of natural resources can be alleviated again, research in recent years shows, Pupa bombycis has considerable value, and it contains the composition such as rich in protein, fatty acid, add and be rich in a large amount of chitin and chitosan, make this " refuse " at food, medicine, and the industry such as chemical industry has purposes widely, can be described as a kind of very abundant and natural resources of preciousness.
2. the mature preparation process of staltic cellulose fibre raw material Cicadae puparium polysaccharide in the present invention
Chitin extraction and then the higher chitosan of preparation purity from Pupa bombycis, can obtain the silkworm chitosan of different de-second phthalein degree by controlling the response time, the highest de-second phthalein degree can reach 93.13%, is suitable as pharma grade product raw material.By the Chitosan powder extracted from Pupa bombycis, to dissolve with gelatin cross-blend and after adding suitable adjuvants, silkworm chitosan compound hemostatic material can be prepared by lyophilization, the sample prepared is spongy, surface and internal void enrich, are evenly distributed, and have certain elastomeric material sense of touch.
3. stanch fibre element coagulating effectiveness of the present invention is good
Silkworm chitosan is the key factor of the coagulant property of compound hemostatic material in this research leading, sample coagulant property effectively improves along with the rising of silkworm chitosan concentration: compared with shrimp and crab shells commodity chitosan, sample coagulating effectiveness prepared by silkworm chitosan is better; Gelatin is added as complexing agent in silkworm chitosan compound hemostatic material, add calcium chloride as coagulant, add glutaraldehyde as cross-linking agent, and add glycerol as moulding dose and antifreeze, all the coagulant property of sample and combination property are served and improve.The silkworm chitosan compound hemostatic material prepared under optimization of C/C composites all has excellent haemostatic effect in vitro and on living animal: compared with hospital gauze, its external clotting index reduces 62.71, make the amount of bleeding in rabbit arteria auricularis Hemorrhage Model decrease 92.08%, bleeding stopping period shortens 80.55%; Compare with medical gelatin sponge group, its external clotting index reduces 35.61%, and make the amount of bleeding in rabbit arteria auricularis Hemorrhage Model decrease 88.35%, bleeding stopping period shortens 57.01%.
4. plasticity is strong
Because the material made by method of the present invention is cellulose fibre, be easy to be processed into various shape, such as spherical (as hemostatic ball), lamellar, plug-like, column, membranaceous, cushion, capsule, bag-shaped etc., not only can be used for plane wound surface, also can be used for cavity-like wound surface, also can with multilayer material and medicine compound make multi-functional take advantage of expect or cohesive stanch bleeding dressing for medical treatment, family stops blooding, takes up an area to give first aid to or burn and protect wound.
5. the material applications made by method of the present invention is extensive
Product of the present invention makes medicine or cosmetics carrier, packaging material.During as pharmaceutical carrier, because many adsorbances are dried in the air greatly in hole, good biocompatibility, degradable easily absorbs, and efficiency is high, and side reaction is little.When the material made by method of the present invention is used as packaging material, solubilized, is easy to process garbage, not only easy to use, and is conducive to environmental conservation; Can be made into non-woven fabrics with the material that method of the present invention is made, also can be made into namely abandoning medicated clothing.
Detailed description of the invention
Below by specific embodiment, the invention will be further described, and following examples are descriptive, is not determinate, can not limit protection scope of the present invention with this.
Example 1
A preparation method for silkworm chitosan compound hemostatic material, the steps include:
(1) prepare silkworm chitosan powder: Pupa bombycis shell is successively through de-inorganic salt, alkali deproteinization, washing and drying, decolouring drying, chitin, de-second phthalein, washing filtering and drying, obtain chitosan, get product after crushed, wherein de-inorganic salt adopts the HCl of 2%, soaking temperature is 30 DEG C, and soak time is 18h; During alkali deproteinization, adopt the NaOH solution of 5% to embathe deproteinising, embathing temperature is 85 DEG C, and the time of embathing is 4h; Employing mass fraction is the H2O2 solution of 20%, and pH=11, treatment temperature are 80 DEG C, and the reason time is 2h, and bath raio is 1:40, and in this step, other operational approach are prior art.
(2) silkworm chitosan powder is put into the glacial acetic acid of 2%, is stirred to and dissolves completely, form the silkworm chitosan solution that mass fraction is 2.5%, then at constant temperature 60 DEG C by mass fraction be 3.0% gelatin be mixed into wherein to be stirred to and dissolve completely;
(3) after being cooled to room temperature, drip successively mass fraction be 0.005% glutaraldehyde solution, mass fraction be 0.1% calcium chloride solution, mass fraction be the glycerol of 0.05%, be uniformly mixed;
(4) the above-mentioned blend solution magnetic stirring apparatus prepared at room temperature is carried out the stirring foaming of 30min, then pour into and be covered with in the culture dish of polyethylene film, schedule to last at pre-freezing temperature in the ultra cold storage freezer of-40 DEG C and carry out 1h pre-freeze, finally put into the dry 30h of frozen vacuum dryer, i.e. obtained silkworm chitosan compound hemostatic material, after ultra violet lamp sterilizing 20min, to preserve under Polythene Bag encapsulation room temperature.
All the other steps are prior art.
Embodiment 2
A preparation method for silkworm chitosan compound hemostatic material, the steps include:
(1) prepare silkworm chitosan powder: with embodiment 1 step (1).
(2) silkworm chitosan powder is put into the glacial acetic acid of 2%, is stirred to and dissolves completely, form the silkworm chitosan solution that mass fraction is 2%, then at constant temperature 60 DEG C by mass fraction be 3.0% gelatin be mixed into wherein to be stirred to and dissolve completely;
(3) after being cooled to room temperature, drip successively mass fraction be 0.010% glutaraldehyde, mass fraction be 0.2% calcium chloride solution, mass fraction be the glycerol of 0.1%, be uniformly mixed;
(4) the above-mentioned blend solution magnetic stirring apparatus prepared at room temperature is carried out the stirring foaming of 30min, then pour into and be covered with in the culture dish of polyethylene film, schedule to last at pre-freezing temperature in the ultra cold storage freezer of-40 DEG C and carry out 1h pre-freeze, finally put into the dry 30h of frozen vacuum dryer, i.e. obtained silkworm chitosan compound hemostatic material, after ultra violet lamp sterilizing 20min, to preserve under Polythene Bag encapsulation room temperature.
Embodiment 3
A preparation method for silkworm chitosan compound hemostatic material, the steps include:
(1) prepare silkworm chitosan powder: with embodiment 1 step (1).
(2) silkworm chitosan powder is put into the glacial acetic acid of 2%, is stirred to and dissolves completely, form the silkworm chitosan solution that mass fraction is 1.5%, then at constant temperature 60 DEG C by mass fraction be 3.0% gelatin be mixed into wherein to be stirred to and dissolve completely;
(3) after being cooled to room temperature, drip successively mass fraction be 0.015% glutaraldehyde, mass fraction be 0.4% calcium chloride solution, mass fraction be the glycerol of 0.15%, be uniformly mixed;
(4) the above-mentioned blend solution magnetic stirring apparatus prepared at room temperature is carried out the stirring foaming of 30min, then pour into and be covered with in the culture dish of polyethylene film, schedule to last at pre-freezing temperature in the ultra cold storage freezer of-40 DEG C and carry out 1h pre-freeze, finally put into the dry 30h of frozen vacuum dryer, i.e. obtained silkworm chitosan compound hemostatic material, after ultra violet lamp sterilizing 20min, to preserve under Polythene Bag encapsulation room temperature.
Embodiment 4
A preparation method for silkworm chitosan compound hemostatic material, the steps include:
(1) prepare silkworm chitosan powder: with embodiment 1 step (1).
(2) silkworm chitosan powder is put into the glacial acetic acid of 2%, is stirred to and dissolves completely, form the silkworm chitosan solution that mass fraction is 1%, then at constant temperature 60 DEG C by mass fraction be 1.0% gelatin be mixed into wherein to be stirred to and dissolve completely;
(3) after being cooled to room temperature, drip successively mass fraction be 0.020% glutaraldehyde, mass fraction be 0.1% calcium chloride solution, mass fraction be the glycerol of 0.1%, be uniformly mixed;
(4) the above-mentioned blend solution magnetic stirring apparatus prepared at room temperature is carried out the stirring foaming of 30min, then pour into and be covered with in the culture dish of polyethylene film, schedule to last at pre-freezing temperature in the ultra cold storage freezer of-40 DEG C and carry out 1h pre-freeze, finally put into the dry 30h of frozen vacuum dryer, i.e. obtained silkworm chitosan compound hemostatic material, after ultra violet lamp sterilizing 20min, to preserve under Polythene Bag encapsulation room temperature.
The wound healing assay of silkworm chitosan compound hemostatic material
Experimental raw is in table 1
Table 1 test raw material and source
| Sequence number | Material name | Raw material sources |
| 1 | Silkworm chitosan compound hemostatic material | Self-control |
| 2 | Hospital gauze | Shandong Wei Guang hygienic material company limited |
| 3 | Rabbit | Female |
Agents useful for same is in table 2
Table 2 reagent name and manufacturer
| Sequence number | Reagent name | Manufacturer |
| 1 | Ether | Tianjin chemical reagent factory |
| 2 | Ethanol | Tianjin chemical reagent factory |
| 3 | Formalin | Tianjin chemical reagent factory |
Experimental technique
Rabbit scalding model makes
Preparatory work of experiment
Select adult healthy Female rabbits 6, quality is 2.5-3.0kg/.Before test, lose hair or feathers in rabbit back spinal both sides by 24h, exposes skin completely, notes not injuring skin shaving in mao process.Be the trial zone of 3cmx3cm at 2 areas that each side select of unhairing district spinal column, respectively as silkworm chitosan compound hemostatic material group and gelfoam matched group trial zone, carry out disinfection with 70% ethanol.Sample is cut into corresponding size for subsequent use: chitosan compound hemostatic material 2cmX2cm; Gauze 3cmX3cm.
Scalding model makes
Adopt open inhalation to anaesthetize tested rabbit before scalding, 5-7 layer hospital gauze is covered on the mouth and nose of rabbit, dropwise drips a small amount of ether slowly on gauze, until rabbit no longer struggles and breathes steadily.
Flat based tubes certain for floor space size is put into 99 DEG C of thermostat water baths to heat, take out after l 0min, the trial zone fitting in rabbit spinal column both sides is immediately scalded, and notes avoiding ilium and shoulder foot bone.Start timing to contact rabbit skin bottom test tube, the scald time is 30s.Two scald wounds are prepared in every rabbit side.
The deposited of sample adds
Each for rabbit side is divided into silkworm chitosan composite materials testing group and blank group at random.Test group scald wound applies in addition silkworm chitosan compound hemostatic material, then covers hospital gauze outward at silkworm chitosan compound hemostatic material and fixed; Fixed after blank group scald wound directly covers hospital gauze.The hyperphoric once deposited sample gauze added every other day.
Wound tissue sections checks
Rabbit scald wound tissue is delivered Tianjin Union Medicine Centre Pathology Deparment, entrusts its pathologic finding carrying out wound tissue sections and take tissue slice figure.
Experimental result and analysis
Wound area and healing rate are in table 4
The wound area of table 4 different time and wound healing rate
Apply as can be seen from Table 4 and added silkworm chitosan compound hemostatic material and do not applied the wound surface area of two groups of scald wounds when 3d, 7d, 14d and 21d and the situation of change of healing rate that add sample.Original wound surface area is scald the area in test bottom test tube, and healing rate is after 0.21d, and apply the wound surface area having added silkworm chitosan compound hemostatic material and narrowed down to about 19mm2, healing rate reaches 96.07%; The deposited blank adding sample is formed face area and is only reduced to about 163mm2, and healing rate is only 67.34%.The wound promoting healing effect of visible silkworm chitosan compound hemostatic material to rabbit scalding model is fairly obvious.
Apply and added silkworm chitosan compound hemostatic material and the deposited situation of change of two groups of scald wound areas along with the time adding sample.After scald in 21d, wound is all in contraction healing, but the deposited speed that Wound Contraction is healed that adds of silkworm chitosan compound hemostatic material is obviously accelerated, and wound area is all less than blank group in each time interval.
Apply and added silkworm chitosan compound hemostatic material and the deposited situation of change of two groups of wound tissue healing rates along with the time adding sample.When 3d, 7d, 14d and 21d, apply added silkworm chitosan compound hemostatic material Wound healing rate all higher than sky from matched group.Blank group is after 7d, and healing rate slows down to some extent, and test group heals, Jie's speed remains on comparatively faster state always.
Claims (6)
1. a preparation method for silkworm chitosan compound hemostatic material, is characterized in that: the steps include:
(1) silkworm chitosan powder is put into the glacial acetic acid of 2%, is stirred to and dissolves completely, form the silkworm chitosan solution that mass fraction is 1.0% ~ 2.5%, then at constant temperature 60 DEG C by mass fraction be 3.0% gelatin be mixed into wherein to be stirred to and dissolve completely;
(2) after being cooled to room temperature, drip successively mass fraction be 0.005% ~ 0.020% glutaraldehyde solution, mass fraction be 0.1% ~ 0.4% calcium chloride solution, mass fraction be the glycerol of 0.05% ~ 0.2%, be uniformly mixed;
(3) the above-mentioned blend solution magnetic stirring apparatus prepared at room temperature is carried out the stirring foaming of 30min, then pour into and be covered with in the culture dish of polyethylene film, schedule to last at pre-freezing temperature in the ultra cold storage freezer of-40 DEG C and carry out 1h pre-freeze, finally put into the dry 30h of frozen vacuum dryer, i.e. obtained silkworm chitosan compound hemostatic material.
2. the preparation method of silkworm chitosan compound hemostatic material according to claim 1, is characterized in that: after described silkworm chitosan compound hemostatic material ultra violet lamp sterilizing 20min, to preserve under Polythene Bag encapsulation room temperature.
3. the preparation method of silkworm chitosan compound hemostatic material according to claim 1, it is characterized in that: by the preparation method of silkworm chitosan powder be: Pupa bombycis shell is successively through de-inorganic salt, alkali deproteinization, washing and drying, decolouring drying, chitin, de-second phthalein, washing filtering and drying, obtain chitosan, get product after crushed.
4. the preparation method of silkworm chitosan compound hemostatic material according to claim 3, is characterized in that: described de-inorganic salt adopts the HCl of 3%, and soaking temperature is 30 DEG C, and soak time is 18h.
5. the preparation method of silkworm chitosan compound hemostatic material according to claim 3, is characterized in that: during described alkali deproteinization, and adopt the NaOH solution of 5% to embathe deproteinising, embathing temperature is 85 DEG C, and the time of embathing is 4h.
6. the preparation method of silkworm chitosan compound hemostatic material according to claim 3, is characterized in that: when described decolouring is dry, and employing mass fraction is the H2O2 solution of 20%, and pH=11, treatment temperature are 80 DEG C, and the reason time is 2h, and bath raio is 1:40.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114949332A (en) * | 2022-04-15 | 2022-08-30 | 永康市第一人民医院 | Treating agent for rapidly promoting wound hemostasis and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114949332A (en) * | 2022-04-15 | 2022-08-30 | 永康市第一人民医院 | Treating agent for rapidly promoting wound hemostasis and preparation method thereof |
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