CN103525097A - Aquatic product fish skin gelatin sponge and preparation method thereof - Google Patents
Aquatic product fish skin gelatin sponge and preparation method thereof Download PDFInfo
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- CN103525097A CN103525097A CN201310503401.4A CN201310503401A CN103525097A CN 103525097 A CN103525097 A CN 103525097A CN 201310503401 A CN201310503401 A CN 201310503401A CN 103525097 A CN103525097 A CN 103525097A
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Abstract
The invention relates to the technical field of medical instruments, and in particular relates to an aquatic product fish skin gelatin sponge and a preparation method thereof. The aquatic product fish skin gelatin is used as a main material, chitosan is used as an auxiliary material, and glutaraldehyde is used as a cross-linking agent; the raw material liquids of different proportions are subjected to freeze drying to prepare fish skin gelatin sponges with different properties; the preparation process of the sponge is simple; the sponge is low in cost and readily available; some defects of the conventional collagen sponge are overcome. The prepared fish skin gelatin sponge has a uniform porous netlike microstructure and higher mechanical strength and good moisture absorption and is a better biomedical material.
Description
Technical field
The present invention relates to technical field of medical instruments, particularly relate to a kind of aquatic products fishskin gelatin sponge and preparation method thereof.
Background technology
Collagen protein is that animal body intensive amount is the abundantest, protein the most widely distributes.Collagen protein has the features such as unique triple-helix structure, low antigenicity, nontoxicity, good biocompatibility and biodegradability, has been widely used in the bio-medical fields such as tissue engineering bracket material, styptic sponge, medicine control slow-released system, gene delivery vector, cosmetic surgery.At present, the collagen protein of domestic use is mainly from skin and the heel string of pig and ox, but along with the generation of the diseases such as mad cow disease and foot and mouth disease, makes people produce query to their security; In addition, due to reasons such as religion and customs, the application of the collagen protein in pig source in certain areas is restricted.Therefore, in the urgent need to seeking the better collagen protein of quality and security source.
The annual fish scale of China, the fishery products wastes such as fish-skin approximately have more than 40 ten thousand tons, these fishery products wastes all contain abundant collagen protein, its amino acid forms and Lu Sheng Mammals collagen protein no significant difference, and there is not the communicable disease of infecting both domestic animals and human in it, have great development and utilization and be worth, but these wastes are used to produce feed except small portion, major part is but dropped.Therefore, develop the new way of utilizing of the fishery products wastes such as fish-skin, not only can improve the added value of processing of aquatic products, and can reduce environmental pollution, there is good economic and social benefit.
Yet as bio-medical material, aquatic products collagen and Lu Sheng Mammals collagen exist common shortcoming, as fast in vivo degradation, snappiness is poor etc., has limited its application.Chitosan is alkaline polysaccharide unique in natural polysaccharide, there is the features such as good biocompatibility, biodegradability, germ resistance, nontoxicity, reduced immunogenicity, water absorbability, flexibility, short blood coagulation, anti, made up to a certain extent the weak point of collagen protein.In addition, by glutaraldehyde, as linking agent, also can increase the degradation rate in the physical strength of collagen protein medical material its body that slows down.
Summary of the invention
The object of the invention is to provide a kind of aquatic products fishskin gelatin sponge and preparation method thereof.
For achieving the above object, the present invention adopts technical scheme to be:
A kind of aquatic products fishskin gelatin sponge:
Aquatic products fishskin gelatin fully dissolves through water-bath, freezing, be drying to obtain aquatic products fishskin gelatin sponge;
Or aquatic products fishskin gelatin fully dissolves through water-bath, after dissolving, add glutaraldehyde cross-linking, freezing, be drying to obtain aquatic products fishskin gelatin sponge;
Or aquatic products fishskin gelatin and chitosan part are fully dissolved through water-bath, dissolve rear mixed freezing, be drying to obtain aquatic products fishskin gelatin sponge;
Or aquatic products fishskin gelatin and chitosan part are fully dissolved through water-bath, after dissolving, mix and add glutaraldehyde cross-linking freezing, be drying to obtain aquatic products fishskin gelatin sponge.
Described aquatic products fishskin gelatin is tilapia fishskin gelatin, and it is triple-helix structure, molecular weight 300kDa left and right, and concentration is 5mg/mL-30mg/mL.
Described glutaraldehyde concentration is 0.5%, the 0.5%-2.5% that add-on is mixeding liquid volume.
Described aquatic products fishskin gelatin is 9:1-5:5 with chitosan mass ratio, and deacetylating degree of chitosan is 85%.
The preparation method of a kind of aquatic products fishskin gelatin sponge:
Aquatic products fishskin gelatin is dissolved in the 30-50 ℃ of acetum under water bath condition, after vacuum outgas 15-30min, in 4 ℃, spend the night crosslinked,-20 ℃ of precooling 8h, then-40--60 ℃ of lyophilize 20-30h, after lyophilize, in 50-60 ℃ of vacuum drying oven, dry 12-20h, remove free acetic acid molecule, obtain aquatic products fishskin gelatin sponge;
Or, aquatic products fishskin gelatin is dissolved in the 30-50 ℃ of acetum under water bath condition, then adding mass concentration is 0.5% glutaraldehyde, after fully mixing, after vacuum outgas 15-30min, in 4 ℃, spend the night crosslinked,-20 ℃ of precooling 8h, then-40--60 ℃ of lyophilize 20-30h dries 12-20h after lyophilize in 50-60 ℃ of vacuum drying oven, remove free glutaraldehyde and acetic acid molecule, obtain aquatic products fishskin gelatin sponge;
Or, aquatic products fishskin gelatin and chitosan are dissolved in respectively in the 30-50 ℃ of acetum under water bath condition, then mixing fishskin gelatin and chitosan total concn in mixed solution is 10mg/mL, after mixed solution vacuum outgas 15-30min, in 4 ℃, spend the night crosslinked ,-20 ℃ of precooling 8h, then-40--60 ℃ of lyophilize 20-30h, after lyophilize, in 50-60 ℃ of vacuum drying oven, dry 12-20h, remove free acetic acid molecule, obtain aquatic products fishskin gelatin sponge;
Or, aquatic products fishskin gelatin and chitosan are dissolved in respectively in the 30-50 ℃ of acetum under water bath condition, then mixing fishskin gelatin and chitosan total concn in mixed solution is 10mg/mL, in mixed solution, adding mass concentration is 0.5% glutaraldehyde again, fully mix after the degassed 15-30min of final vacuum, in 4 ℃, spend the night crosslinked,-20 ℃ of precooling 8h, then-40--60 ℃ of lyophilize 20-30h, after lyophilize, in 50-60 ℃ of vacuum drying oven, dry 12-20h, remove free glutaraldehyde and acetic acid molecule, obtain aquatic products fishskin gelatin sponge.
Described aquatic products fishskin gelatin is tilapia fishskin gelatin, and it is triple-helix structure, molecular weight 300kDa left and right.
Described aquatic products fishskin gelatin is 9:1-5:5 with chitosan mass ratio.
Further, aquatic products fishskin gelatin and chitosan are dissolved in respectively in 0.05mol/L acetum, in 50 ℃ of water-baths, fully dissolve respectively, after dissolving, chitosan-acetic acid solution is slowly joined in aquatic products fishskin gelatin acetum, after mixing, 0.5% glutaraldehyde cross-linking that adds 1% mixeding liquid volume under magnetic agitation condition, after stirring and evenly mixing by mixed solution vacuum outgas 20min, pour in mould, 4 ℃ are spent the night crosslinked,-20 ℃ of precooling 8h, then-60 ℃ of lyophilize 20h, take out in type sponge product, in 60 ℃ of vacuum drying ovens, dry 12h, remove free glutaraldehyde and acetic acid molecule, obtain aquatic products fishskin gelatin sponge.
The present invention has advantages of: preparation process of the present invention is simple, cheap and easy to get, has overcome the some shortcomings of existing collagen sponge.Fishskin gelatin sponge prepared by the present invention, its microtexture is uniform holey, has stronger physical strength and good water absorbability, is a kind of good bio-medical material.Particular embodiment,
1. the present invention's gelatin used is aquatic products waste material Java tilapia skin extract, raw material sources safety, and the present invention makes full use of the fish-skin waste material producing in the tilapia aquatic products course of processing, turns waste into wealth, and increases economic efficiency.
2. to adopt raw material be to adopt hot-water extraction method from tilapia fishskin, to extract the gelatin obtaining in the present invention, but not the collagen protein that enzymolysis process obtains, its extracting method is easier, economy and gelatin thermostability higher; Meanwhile, the raw material in the present invention is the Java tilapia skin gelatin dried product of spraying, more easy and economical in transportation, storage and the use procedure of raw material.
3. the present invention can develop Mammals (pig and ox) with the aquatic products collagen sponge of external source, and its safety coefficient is higher, output is larger.
4. although the present invention is to be compared with prior art that major ingredient, chitosan auxiliary material and glutaraldehyde as cross linker are produced sponge with the raw material that aquatic products fish-skin extracts, but the proportioning of prepared sponge is all not identical, and the present invention can obtain the gelfoam with glutaraldehyde cross-linking, also can obtain the gelfoam of being made by Java tilapia skin gelatin separately, its safety coefficient is higher, subsequent disposal is simpler; Chemical cross-linking agent of the present invention only has glutaraldehyde simultaneously, and main formula material is Java tilapia skin gelatin and a small amount of chitosan, does not use other chemical feedstockss.
5. the sponge of the resulting a series of different proportionings of the present invention, its mode of appearance is attractive in appearance, texture is soft is more easily accepted by human consumer; And its fiber thickness is even, pore size is moderate and even; Ventilation property and water absorbability are good, physical strength is moderate, snappiness is better; Simultaneously from microtexture, fiber thickness more evenly, pore size is more moderate, and pore distribution is more even, is mostly to communicate between Kong Yukong, and this is more conducive to breathe freely.
Accompanying drawing explanation
The mode of appearance figure of several different proportioning gelfoams that Fig. 1-6 provide for the embodiment of the present invention; Wherein, Fig. 1 proportioning is 10mg/mL gelatin; Fig. 2 proportioning is 0.5% glutaraldehyde of 10mg/mL gelatin and 1% volume; Fig. 3 proportioning is that gelatin and chitosan ratio are 9:1, and total concn is 10mg/mL; Fig. 4 proportioning is that gelatin and chitosan ratio are 8:2, and total concn is 10mg/mL; Fig. 5 proportioning is that gelatin and chitosan ratio are 8:2, and total concn is 0.5% glutaraldehyde of 10mg/mL and 1% volume; Fig. 6 proportioning is that gelatin and chitosan ratio are 9:1, and total concn is 0.5% glutaraldehyde of 10mg/mL and 2% volume.
The plane microtexture of several different proportioning gelfoams that Fig. 7-12 provide for the embodiment of the present invention; Wherein, Fig. 7 proportioning is 10mg/mL gelatin; Fig. 8 proportioning is 0.5% glutaraldehyde of 10mg/mL gelatin and 1% volume; Fig. 9 proportioning is that gelatin and chitosan ratio are 9:1, and total concn is 10mg/mL; Figure 10 proportioning is that gelatin and chitosan ratio are 8:2, and total concn is 10mg/mL; Figure 11 proportioning is that gelatin and chitosan ratio are 8:2, and total concn is 0.5% glutaraldehyde of 10mg/mL and 1% volume; Figure 12 proportioning is that gelatin and chitosan ratio are 9:1, and total concn is 0.5% glutaraldehyde of 10mg/mL and 2% volume.
The section microtexture of several different proportioning gelfoams that Figure 13-18 provide for the embodiment of the present invention; Wherein, Figure 13 proportioning is 10mg/mL gelatin; Figure 14 proportioning is 0.5% glutaraldehyde of 10mg/mL gelatin and 1% volume; Figure 15 proportioning is that gelatin and chitosan ratio are 9:1, and total concn is 10mg/mL; Figure 16 proportioning is that gelatin and chitosan ratio are 8:2, and total concn is 10mg/mL; Figure 17 proportioning is that gelatin and chitosan ratio are 8:2, and total concn is 0.5% glutaraldehyde of 10mg/mL and 1% volume; Figure 18 proportioning is that gelatin and chitosan ratio are 9:1, and total concn is 0.5% glutaraldehyde of 10mg/mL and 2% volume.
Embodiment
With specific embodiment, further illustrate essentiality content of the present invention below, but content of the present invention is not limited to this.
The preparation of embodiment 1 aquatic products fishskin gelatin sponge
The preparation of 1.1 different concns aquatic products fishskin gelatin sponges:
Take the spray-dried Java tilapia skin aquatic products gelatin obtaining of 75mg, 150mg, 225mg, 300mg, 375mg and 450mg, join respectively in 0.05mol/L acetum, constant volume is 15mL, both obtained the gelatin acetum that gelatin concentration is respectively 5mg/mL, 10mg/mL, 15mg/mL, 20mg/mL, 25mg/mL and 30mg/mL, under room temperature suitably after swelling, put into 50 ℃ of water-bath hydrotropies, every 10min, take out to be placed on during this time and on agitator, stir 5min, until gelatin is dissolved in acetum completely, till liquid becomes transparent even shape.After vacuum outgas 20min, pour gelatin solution into 7.5cm * 1.5cm(diameter * height) in culture dish, after preservative film sealing, 4 ℃ are spent the night crosslinked,-20 ℃ of precooling 8h, then-60 ℃ of lyophilize 20h, take out in type sponge product, in 60 ℃ of vacuum drying ovens, dry 12h, remove free acetic acid molecule.Both above-mentioned different concns aquatic products fishskin gelatin sponge (being 10mg/mL gelatin referring to Fig. 1).
The preparation of the aquatic products fishskin gelatin sponge of 1.2 different amount glutaraldehyde cross-linkings:
Take 5 parts of spray-dried Java tilapia skin aquatic products gelatin that obtain, every part of 150mg, join respectively in 0.05mol/L acetum, constant volume is 15mL, both the concentration gelatin acetum that is 10mg/mL, under room temperature suitably after swelling, put into 50 ℃ of water-bath hydrotropies, 10min takes out to be placed on and on agitator, stirs 5min during this time, until gelatin is dissolved in acetum completely, till liquid becomes transparent even shape.Under the condition constantly stirring at magnetic stirring apparatus, in gelatin solution, add respectively 0.075mL (0.5%), 0.15mL (1%), 0.225mL(1.5%), 0.3mL (2%) and 0.5% glutaraldehyde 0.375mL(2.5%), fully after stirring and evenly mixing, after vacuum outgas 20min, pour mixed solution into 7.5cm * 1.5cm(diameter * height) in culture dish, after preservative film sealing, 4 ℃ are spent the night crosslinked,-20 ℃ of precooling 8h, then-60 ℃ of lyophilize 20h, take out in type sponge product, in 60 ℃ of vacuum drying ovens, dry 12h, remove free glutaraldehyde and acetic acid molecule.Both the crosslinked aquatic products fishskin gelatin sponge (being 0.5% glutaraldehyde of 10mg/mL gelatin and 1% volume referring to Fig. 2) of different glutaraldehyde amounts.
1.3 different aquatic products fishskin gelatins and chitosan mass are than the preparation of sponge:
Take the spray-dried Java tilapia skin aquatic products gelatin obtaining of 135mg and 15mg chitosan (mass ratio 9:1 between fishskin gelatin and chitosan), join respectively in 10mL and 5mL0.05mol/L acetum, under room temperature suitably after swelling, put into 50 ℃ of water-bath hydrotropies, 10min takes out to be placed on and on agitator, stirs 5min during this time, until gelatin and chitosan be dissolved in acetum completely, till liquid becomes transparent even shape.Under the condition constantly stirring at magnetic stirring apparatus, chitosan solution is slowly joined in gelatin solution, fully after stirring and evenly mixing, after vacuum outgas 20min, pour mixed solution into 7.5cm * 1.5cm(diameter * height) in culture dish, after preservative film sealing, 4 ℃ spend the night crosslinked ,-20 ℃ of precooling 8h, then-60 ℃ of lyophilize 20h, take out in type sponge product, in 60 ℃ of vacuum drying ovens, dry 12h, remove free acetic acid molecule.Both for the aquatic products fishskin gelatin sponge of 9:1, (referring to Fig. 3, be that gelatin and chitosan ratio are 9:1, total concn is 10mg/mL with chitosan mass ratio to have obtained aquatic products fishskin gelatin; Fig. 4 is that gelatin and chitosan ratio are 8:2, and total concn is 10mg/mL).
Then according to aforesaid way, obtain respectively aquatic products fishskin gelatin and chitosan mass than being the aquatic products fishskin gelatin sponge of 8:2,7:3,6:4 and 5:5.
The preparation of the gelfoam of 1.4 different aquatic products fishskin gelatins, chitosan and glutaraldehyde proportioning:
In above-mentioned 1.1,1.2 and 1.3, test, can determine suitable single factor, that is: aquatic products fishskin gelatin concentration is 10mg/mL, and glutaraldehyde add-on is 1%, and gelatin than being 8:2, is designed suitable level of factor table with chitosan mass thus, and result is as follows:
Level of factor table
By quadrature software, draw orthogonal design table, as follows:
Orthogonal design table
Note: the final volume of each group experiment is 15mL.
With reference to orthogonal design table, after each group experimental liquid is mixed, after vacuum outgas 20min, pour mixed solution into 7.5cm * 1.5cm(diameter * height) in culture dish, after preservative film sealing, 4 ℃ are spent the night crosslinked,-20 ℃ of precooling 8h, then-60 ℃ of lyophilize 20h, take out in type sponge product, in 60 ℃ of vacuum drying ovens, dry 12h, remove free glutaraldehyde and acetic acid molecule.Both the gelfoam of different aquatic products fishskin gelatins, chitosan and glutaraldehyde proportioning (referring to Fig. 5 gelatin and chitosan, than being 8:2, total concn is 0.5% glutaraldehyde of 10mg/mL and 1% volume; Fig. 6 gelatin and chitosan are than being 9:1, and total concn is 0.5% glutaraldehyde of 10mg/mL and 2% volume).
Embodiment 2
The mensuration of aquatic products fishskin gelatin sponge character:
Choose each gelfoam product that the good above-described embodiment of mode of appearance obtains and carry out scanning electron microscope detection, observe the thickness of its fiber, (referring to Fig. 7-18) such as degree of uniformity in the size in aperture, aperture.
By scanning electron microscope picture, can be found out, when amplifying 100 times:
1. its hole of sponge of being made by Java tilapia skin gelatin is separately large, fiber is thin, space is not very even;
2. the sponge space of being made by Java tilapia skin gelatin and glutaraldehyde is less, fiber is thick, space is very even;
3. the sponge space of being made by Luo Fei fishskin gelatin and chitosan (9:1 and 8:2) is less, fiber is thick, space is even, and the sponge space that the 8:2 sponge of making is made than 9:1 is more even;
4. the sponge space of being made by Luo Fei fishskin gelatin, chitosan and glutaraldehyde is very little, fiber is very thick but space is even.
The mensuration of aquatic products fishskin gelatin sponge hygroscopic nature:
Each fishskin gelatin sponge that above-described embodiment is obtained is measured accordingly, and W specifically weighs the gelfoam of oven dry
1, then gelfoam is put into pure water, allow its 2min that naturally absorbs water, with skimming ladle, the sponge of fully water suction to be pulled out, drop is its surface-moisture to the greatest extent, about 30s, rapid weighing W
2, the water suction multiple of calculating gelfoam product, formula is as follows:
Water suction multiple=(W2-W1)/W1
After measured, proportioning is 10mg/mL gelatin; Proportioning is 0.5% glutaraldehyde of 10mg/mL gelatin and 1% volume; Proportioning is that gelatin and chitosan ratio are 9:1, and total concn is 10mg/mL; Proportioning is that gelatin and chitosan ratio are 8:2, and total concn is 10mg/mL; Proportioning is that gelatin and chitosan ratio are 8:2, and total concn is 0.5% glutaraldehyde of 10mg/mL and 1% volume; Proportioning be gelatin and chitosan than being 9:1, total concn is that the water suction multiple of the made gelfoam product of 0.5% glutaraldehyde of 10mg/mL and 2% volume is all in 32-45 left and right doubly.
The ventilative property testing of aquatic products fishskin gelatin sponge:
Each fishskin gelatin sponge that above-described embodiment is obtained is measured accordingly, specifically in the identical 50mL Erlenmeyer flask of two sizes, adds the pure water of same amount, about 20mL, and accurately weigh is designated as W respectively
1and W
2, aquatic products fishskin gelatin sponge is cut into Erlenmeyer flask bottleneck circle of the same size and is encapsulated in bottleneck, bottleneck is around by the sealed membrane reality of obturaging, control group open-ended, all be placed in 24h in same glass moisture eliminator by two bottles, make moisture spontaneous evaporation, the weight that then accurately weighs experimental group and control group is designated as W
1' and W
2'.Calculate gelfoam product ventilation property, formula is as follows:
Air penetrability=(W1-W1 ')/(W2-W2 ')
After measured, said ratio is 10mg/mL gelatin; Proportioning is 0.5% glutaraldehyde of 10mg/mL gelatin and 1% volume; Proportioning is that gelatin and chitosan ratio are 9:1, and total concn is 10mg/mL; Proportioning is that gelatin and chitosan ratio are 8:2, and total concn is 10mg/mL; Proportioning is that gelatin and chitosan ratio are 8:2, total concn be 0.5% glutaraldehyde of 10mg/mL and 1% volume and proportioning be gelatin and chitosan than being 9:1, total concn is that the air penetrability of the made gelfoam product of 0.5% glutaraldehyde of 10mg/mL and 2% volume is all in 42%-49% left and right.
The mensuration of aquatic products fishskin gelatin sponge moisturizing character:
Each fishskin gelatin sponge that above-described embodiment is obtained is measured accordingly, the aquatic products gelfoam that concrete clip the is of moderate size W that weighs
1, then gelfoam is put into pure water, allow its 2min that naturally absorbs water, with skimming ladle, the sponge of fully water suction to be pulled out, drop is its surface-moisture to the greatest extent, and about 30s, weighs its weight W
2, then sponge is placed in to the special centrifuge tube with filter membrane, 25 ℃, the centrifugal 5min of 500r/min, takes out the sponge W that again weighs
3.Calculate gelfoam moisture retention, formula is as follows:
Moisturizing rate=(W
2-W
1)/(W
3-W
1)
After measured, proportioning is 10mg/mL gelatin; Proportioning is 0.5% glutaraldehyde of 10mg/mL gelatin and 1% volume; Proportioning is that gelatin and chitosan ratio are 9:1, and total concn is 10mg/mL; Proportioning is that gelatin and chitosan ratio are 8:2, and total concn is 10mg/mL; Proportioning is that gelatin and chitosan ratio are 8:2, and total concn is 0.5% glutaraldehyde of 10mg/mL and 1% volume; Proportioning be gelatin and chitosan than being 9:1, total concn is that the moisturizing rate of the made gelfoam product of 0.5% glutaraldehyde of 10mg/mL and 2% volume is all in 22-39 left and right doubly.
The mensuration of aquatic products fishskin gelatin sponge stretching resistance character:
Each fishskin gelatin sponge that above-described embodiment is obtained is measured accordingly, specifically gelfoam is trimmed to square, calculate the area S of section, with glue, fixedly one end of sponge is on wooden unit, and the other end, on plank, applies one perpendicular to the pulling force of plank at sponge plank end, till slowly increasing to sponge fracture, tensile force f during record fracture, the stretching resistance character of calculating gelfoam product, formula is as follows:
Tensile strength (N/cm
2)=F/S
After measured, proportioning is 10mg/mL gelatin; Proportioning is 0.5% glutaraldehyde of 10mg/mL gelatin and 1% volume; Proportioning is that gelatin and chitosan ratio are 9:1, and total concn is 10mg/mL; Proportioning is that gelatin and chitosan ratio are 8:2, and total concn is 10mg/mL; Proportioning is that gelatin and chitosan ratio are 8:2, total concn be 0.5% glutaraldehyde of 10mg/mL and 1% volume and proportioning be gelatin and chitosan than being 9:1, total concn is that the Tensile strength of the made gelfoam product of 0.5% glutaraldehyde of 10mg/mL+2% volume is all in 0.4-0.9 left and right.
Degrade the outward mensuration of character of aquatic products fishskin gelatin cavernous body:
Each fishskin gelatin sponge that above-described embodiment is obtained is measured accordingly, the concrete gelfoam product that takes respectively 5mg gelatin and different ingredients is in 5mL centrifuge tube, add 1mL0.1M PBS(pH7.4) immersion 1h, under 37 ℃ of water-baths, add 0.05MCaCl2 and 200U collagenase, place respectively 12h, 24h, 36h, 48h, 60h and 72h, then centrifuge tube being put into ice-water bath adds 0.2mL0.25M EDTA solution to make reaction terminating, centrifugal 15min under 4 ℃ of 5000r/min, take out 1mL supernatant liquor in ampoule, the hydrochloric acid that adds 1mL6M, with being hydrolyzed 3h after alcohol blast burner sealing at 130 ℃.Take out cooling, with distilled water, be settled to 10mL, get 1mL sample liquid and add each reagent by the measuring method of oxyproline typical curve, after blank zeroing, measure the light absorption value at each sample liquid 560nm place, after converting, draw the amount of the gelatin being hydrolyzed by collagenase, calculate gelatin product external degradation rate, formula is as follows:
Degradation rate (%)=W
1/ W
2
W1: the gelatin amount being hydrolyzed
W2: the gelatin amount containing in gelatin product
The results are shown in following table
Claims (8)
1. an aquatic products fishskin gelatin sponge, is characterized in that:
Aquatic products fishskin gelatin fully dissolves through water-bath, freezing, be drying to obtain aquatic products fishskin gelatin sponge;
Or aquatic products fishskin gelatin fully dissolves through water-bath, after dissolving, add glutaraldehyde cross-linking, freezing, be drying to obtain aquatic products fishskin gelatin sponge;
Or aquatic products fishskin gelatin and chitosan part are fully dissolved through water-bath, dissolve rear mixed freezing, be drying to obtain aquatic products fishskin gelatin sponge;
Or aquatic products fishskin gelatin and chitosan part are fully dissolved through water-bath, after dissolving, mix and add glutaraldehyde cross-linking freezing, be drying to obtain aquatic products fishskin gelatin sponge.
2. by aquatic products fishskin gelatin sponge claimed in claim 1, it is characterized in that: described aquatic products fishskin gelatin is tilapia fishskin gelatin, it is triple-helix structure, molecular weight 300kDa left and right, and concentration is 5mg/mL-30mg/mL.
3. by aquatic products fishskin gelatin sponge claimed in claim 1, it is characterized in that: telling glutaraldehyde concentration is 0.5%, the 0.5%-2.5% that add-on is mixeding liquid volume.
4. by aquatic products fishskin gelatin sponge claimed in claim 1, it is characterized in that: described aquatic products fishskin gelatin is 9:1-5:5 with chitosan mass ratio, and deacetylating degree of chitosan is 85%.
5. a preparation method for aquatic products fishskin gelatin sponge claimed in claim 1, is characterized in that:
Aquatic products fishskin gelatin is dissolved in the 30-50 ℃ of acetum under water bath condition, after vacuum outgas 15-30min, in 4 ℃, spend the night crosslinked,-20 ℃ of precooling 8h, then-40--60 ℃ of lyophilize 20-30h, after lyophilize, in 50-60 ℃ of vacuum drying oven, dry 12-20h, remove free acetic acid molecule, obtain aquatic products fishskin gelatin sponge;
Or, aquatic products fishskin gelatin is dissolved in the 30-50 ℃ of acetum under water bath condition, then adding mass concentration is 0.5% glutaraldehyde, after fully mixing, after vacuum outgas 15-30min, in 4 ℃, spend the night crosslinked,-20 ℃ of precooling 8h, then-40--60 ℃ of lyophilize 20-30h dries 12-20h after lyophilize in 50-60 ℃ of vacuum drying oven, remove free glutaraldehyde and acetic acid molecule, obtain aquatic products fishskin gelatin sponge;
Or, aquatic products fishskin gelatin and chitosan are dissolved in respectively in the 30-50 ℃ of acetum under water bath condition, then mixing fishskin gelatin and chitosan total concn in mixed solution is 10mg/mL, after mixed solution vacuum outgas 15-30min, in 4 ℃, spend the night crosslinked ,-20 ℃ of precooling 8h, then-40--60 ℃ of lyophilize 20-30h, after lyophilize, in 50-60 ℃ of vacuum drying oven, dry 12-20h, remove free acetic acid molecule, obtain aquatic products fishskin gelatin sponge;
Or, aquatic products fishskin gelatin and chitosan are dissolved in respectively in the 30-50 ℃ of acetum under water bath condition, then mixing fishskin gelatin and chitosan total concn in mixed solution is 10mg/mL, in mixed solution, adding mass concentration is 0.5% glutaraldehyde again, fully mix after the degassed 15-30min of final vacuum, in 4 ℃, spend the night crosslinked,-20 ℃ of precooling 8h, then-40--60 ℃ of lyophilize 20-30h, after lyophilize, in 50-60 ℃ of vacuum drying oven, dry 12-20h, remove free glutaraldehyde and acetic acid molecule, obtain aquatic products fishskin gelatin sponge.
6. by the preparation method of aquatic products fishskin gelatin sponge claimed in claim 4, it is characterized in that: described aquatic products fishskin gelatin is tilapia fishskin gelatin, it is triple-helix structure, molecular weight 300kDa left and right, and concentration is 5mg/mL-30mg/mL.
7. by the preparation method of aquatic products fishskin gelatin sponge claimed in claim 4, it is characterized in that: described aquatic products fishskin gelatin is 9:1-5:5 with chitosan mass ratio, and deacetylating degree of chitosan is 85%.
8. by the preparation method of aquatic products fishskin gelatin sponge claimed in claim 4, it is characterized in that: aquatic products fishskin gelatin and chitosan are dissolved in respectively in 0.05mol/L acetum, in 50 ℃ of water-baths, fully dissolve respectively, after dissolving, chitosan-acetic acid solution is slowly joined in aquatic products fishskin gelatin acetum, after mixing, 0.5% glutaraldehyde cross-linking that adds 1% mixeding liquid volume under magnetic agitation condition, after stirring and evenly mixing by mixed solution vacuum outgas 20min, pour in mould, 4 ℃ are spent the night crosslinked,-20 ℃ of precooling 8h, then-60 ℃ of lyophilize 20h, take out in type sponge product, in 60 ℃ of vacuum drying ovens, dry 12h, remove free glutaraldehyde and acetic acid molecule, obtain aquatic products fishskin gelatin sponge.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105400214A (en) * | 2015-11-30 | 2016-03-16 | 北京化工大学 | Method for preparing hemostatic cotton through hyaluronic acid crosslinked gelatin |
| CN106110379A (en) * | 2016-07-21 | 2016-11-16 | 王雅 | A kind of preparation method of collagen protein sponge |
| IT201700022625A1 (en) * | 2017-02-28 | 2018-08-28 | Consiglio Nazionale Ricerche | FILTER FOR THE EXCHANGE OF HEAT AND HUMIDITY BY APPLICATION IN MEDICAL FIELD AND PROCEDURE FOR ITS PRODUCTION |
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| CN107349912A (en) * | 2017-08-10 | 2017-11-17 | 武汉科技大学 | A kind of carbon composite gelatin sponge and preparation method thereof |
| CN109438774A (en) * | 2018-09-30 | 2019-03-08 | 浙江工业大学 | A kind of preparation method of the natural aromatic sponge of efficient absorption |
| CN113318095A (en) * | 2021-05-17 | 2021-08-31 | 海南海灵化学制药有限公司 | Magnesium sulfate external preparation and preparation method thereof |
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