CN108310449A - A kind of preparation method of gelfoam for hemostasis - Google Patents
A kind of preparation method of gelfoam for hemostasis Download PDFInfo
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Abstract
The invention discloses a kind of preparation methods of the gelfoam for hemostasis, belong to field of medicaments.The present invention, which thaws jellyfish to be crushed, is added papain hydrolysis, then enzyme deactivation filtering, obtains filtrate；Filtrate is centrifuged, takes supernatant that ethyl alcohol alcohol precipitation is added, is centrifuged, filters, obtains filter cake, filter cake ethyl alcohol, acetone alternating elution are dried, dried object is obtained；By gelatin, distilled water immersion, stirring adds dried object, crosslinking agent stirring, obtains stirring liquid；Extract, polycaprolactone, dichloromethane are mixed, sonic oscillation, obtain oscillation liquid, oscillation liquid, polyvinyl alcohol water solution are stirred, is centrifuged, is obtained sediment, sediment is washed, vacuum freezedrying obtains dried object a；Liquid, dried object a Hybrid Heatings will be stirred, stands, obtains mixed liquor, mixed liquor, sodium hydroxide, extracting solution are mixed, got blisters, is freezed, re-dry is cut into small pieces, is obturaged with paper wrapper, and hot air sterilization is sealed in polybag to get the gelfoam for hemostasis.
Description
Technical field
The invention belongs to field of medicaments, and in particular to a kind of preparation method of the gelfoam of hemostasis.
Background technology
Gelatin has good bioactivity, has extremely extensive purposes in biomedical sector.Gelfoam meets
Pharmacopeia has excellent haemostatic properties, is a kind of hemostat that sterile, apyrogeneity can be absorbed by tissue.It can be allowed to stay
It need not take out in human body, absorb naturally.Main component：Protein content accounts for 95% or more.Suitable for various surgical hemostasis, have
Wound is clogged, the function of healing is promoted.Such as：Neurosurgery, the department of stomatology, ear-nose-throat department, gynaecology etc., it is all suitable when need to stop blooding
With.In use, inner packing should be opened under tight aseptic condition, it is cut into the shape of needs, is applied after gently rubbing, or immerses life
It manages in brine, gently rubs after being allowed to drenched, squeeze liquid to the greatest extent, spread at bleeding, until pressing waits for blood clotting.The concrete specification can be by
Break forth blood area selection.A large amount of moisture and diffusate can be quickly absorbed, the dressing of operative incision, the various surface of a wound is can be used as,
And the suppository of various interventional treatment.
At present traditional gelfoam or gelatine-chitosan sponge be mostly with hydrochloric acid, acetate dissolution chitosan, with formaldehyde,
The crosslinking agents such as glutaraldehyde make gelatin or gelatin/chitosan mixed liquor be crosslinked, and finally obtain sponge with freeze-drying dehydration, sterilizing
Shape product.But current gelfoam only plays simple anastalsis, without any anti-infectious function.Therefore, one is produced
The gelfoam of kind higher performance is a necessary thing.
Invention content
The technical problems to be solved by the invention：It is not ideal enough for the haemostatic effect of current gelfoam, it is no any anti-
The problem of infection effect, provides a kind of preparation method of the gelfoam for hemostasis.
In order to solve the above technical problems, the present invention is using technical solution as described below：
The preparation method of a kind of gelfoam for hemostasis, which is characterized in that the preparation method includes the following steps：
（1）Jellyfish thaw broken, is placed in container, the papain of jellyfish quality 0.01 ~ 0.02% is added, in pH
Value is 5 ~ 7, is hydrolyzed at 60 ~ 75 DEG C of temperature, then the enzyme deactivation in the water-bath of 98 ~ 100 DEG C of temperature, is filtered, is obtained with double gauze
Filtrate；
（2）Filtrate is centrifuged, supernatant is taken, the ethyl alcohol that 1 ~ 2 times of supernatant quality is added into supernatant carries out alcohol precipitation, from
The heart filters, obtains filter cake, dry at 30 ~ 40 DEG C of temperature by filter cake ethyl alcohol, acetone alternating elution, obtains dried object；
（3）Count in parts by weight, take 30 ~ 40 parts of distilled water, 10 ~ 15 parts of gelatin, 5 ~ 6 parts of dried objects, 3 ~ 4 parts of extracting solutions, 1 ~ 2 part
Crosslinking agent mixes gelatin, distilled water, impregnates 1 ~ 2h, and extracting solution is added, uniform in 50 ~ 55 DEG C of stirred in water bath of temperature,
Dried object, crosslinking agent are added, is stirred evenly, stirring liquid is obtained, it is spare；
（4）In mass ratio 1：1：3 ~ 4, extract, polycaprolactone, dichloromethane are mixed, sonic oscillation, obtains oscillation liquid,
In mass ratio 1：10 ~ 15, by oscillation liquid, 2 ~ 3% polyvinyl alcohol water solution of mass fraction, stirring, then closed stirring, centrifugation divide
From obtaining sediment, by precipitate washed with water, vacuum freezedrying obtains dried object a；
（5）In mass ratio 10 ~ 15：1, stirring liquid, dried object a are mixed, are heated to 38 ~ 50 DEG C, stands, obtains mixed liquor,
In mass ratio 10：4：0.1, mixed liquor, sodium hydroxide, extracting solution are mixed, got blisters, is freezed in temperature -20 ~ -10 DEG C, then
In 30 ~ 38 DEG C of heated-air dryings of temperature, it is cut into small pieces, is obturaged with paper wrapper, then in 100 ~ 120 DEG C of hot air sterilizations of temperature, is sealed in
To get the gelfoam for hemostasis in polybag.
The step（3）Middle extracting solution is that the rhizome of cogongrass is dried, and crushes, obtains crushed material, in mass ratio 1：7~
8, crushed material, the ethyl alcohol that mass fraction is 80% are mixed, the refluxing extraction at 70 ~ 80 DEG C of temperature, filtering, separating filtrate,
Filter residue, 2 ~ 3 times repeatedly, merging filtrate is to get extracting solution.
The step（3）Middle crosslinking agent is glutaraldehyde.
The step（5）Middle extracting solution is to be ground grape branch, obtains powder, in mass ratio 1：10, by powder,
0.5mol/L ethanol solutions are mixed, in 40 ~ 60 DEG C of temperature, ultrasonic frequency carried out under the conditions of being 100 Hz extraction 15 ~
20min is to get extracting solution.
Compared with other methods, advantageous effects are the present invention：
（1）A kind of present invention polysaccharose substance from jellyfish can significantly affect proliferation, the migration of cell, promote wound repair
And regeneration, and gelatin cross-blend, since gelatin belongs to protein substance, and there may be hydrogen together with protein and carbohydrate
Key acts on and electrostatic interaction, adds crosslinked action, is cross-linked with each other between amido, hydroxyl etc., it is strong to further increase its mechanics
Degree, breaks through sponge, then mixed with polycaprolactone to more efficiently prevent from blood, since polycaprolactone has good life
Object compatibility, degradability and catabolite nontoxicity, good mechanical mechanics property, and can be dissolved out from cavernous body,
Skin surface forms a kind of polycaprolactone film, and due to the presence of film, and a physics is formd between the surface of a wound and subcutaneous tissue
Barrier efficiently reduces the adhesion of the surface of a wound and surrounding tissue, also helps surface of a wound recovery, while gelatin is copolymerized with caprolactone, from
And the degradation rate for even controlling copolymerization product can be improved, absorption of the different pharmaceutical carrier in human body is adapted to, to effectively
Improve the haemostatic effect of gelfoam.
（2）The present invention adsorbs extraction hemostasis class in a kind of rhizome from cogongrass also using polycaprolactone as carrier material
Substance, and hemostasis substance is wrapped, achieve the effect that slow release, to further increase stopping for gelfoam
Blood effect is additionally added a kind of polyphenols extracted from grape branch, to vermin and fungi, bacterium even disease
The infection of poison has good active force, and can do rapid to pathogen etc. and effectively handle, to enable gelfoam
With certain disinfection anti-corrosion, anti-infectious effect.
Specific implementation mode
Extract is that the rhizome of cogongrass is dried, and crushes, obtains crushed material, in mass ratio 1：7 ~ 8, by crushed material, matter
The ethyl alcohol that amount score is 80% is mixed, the refluxing extraction at 70 ~ 80 DEG C of temperature, filtering, separating filtrate, filter residue, and repeatedly 2 ~ 3
Secondary, merging filtrate obtains total filtrate, and total filtrate is concentrated to get extract.
Crosslinking agent is glutaraldehyde.
Extracting solution is to be ground grape branch, obtains powder, in mass ratio 1：10, powder, 0.5mol/L ethyl alcohol is molten
Liquid is mixed, and in 40 ~ 60 DEG C of temperature, ultrasonic frequency carries out 15 ~ 20min of extraction to get extracting solution under the conditions of being 100 Hz.
A kind of preparation method of gelfoam for hemostasis, the preparation method include the following steps：
（1）Jellyfish thaw broken, is placed in container, the papain of jellyfish quality 0.01 ~ 0.02% is added, in pH
Value is 5 ~ 7, is hydrolyzed 1 ~ 2h at 60 ~ 75 DEG C of temperature, then 8 ~ 10min of enzyme deactivation in the water-bath of 98 ~ 100 DEG C of temperature, with bilayer
Filtered through gauze obtains filtrate；
（2）Filtrate is centrifuged into 5 ~ 6min with 800 ~ 1000r/min, supernatant is taken, 1 ~ 2 times of supernatant quality is added into supernatant
Ethyl alcohol carry out alcohol precipitation 20 ~ for 24 hours, 8 ~ 10min is centrifuged with 2000 ~ 3000r/min, filters, filter cake is obtained, by filter cake ethyl alcohol, third
Ketone alternating elution, it is dry at 30 ~ 40 DEG C of temperature, obtain dried object；
（3）It counts in parts by weight, takes 30 ~ 40 parts of distilled water, 10 ~ 15 parts of gelatin, 3 ~ 4 parts of extracting solutions, 1 ~ 2 part of crosslinking agent, it will be bright
Glue, distilled water are mixed, and are impregnated 1 ~ 2h, then uniform in 50 ~ 55 DEG C of stirred in water bath of temperature, are added dried object, crosslinking agent,
It stirs evenly, obtains stirring liquid, it is spare；
（4）In mass ratio 1：1：3 ~ 4, extract, polycaprolactone, dichloromethane are mixed, 25 ~ 30min of sonic oscillation is obtained
Oscillation liquid, in mass ratio 1：10 ~ 15, it will vibrate in liquid, 2 ~ 3% polyvinyl alcohol water solution of mass fraction, with 800 ~ 900r/min
25 ~ 30min is stirred, then with 100 ~ 200r/min, 2 ~ 3h of closed stirring, centrifugation obtains sediment, by precipitate washed with water,
Vacuum freezedrying obtains dried object a；
（5）In mass ratio 10 ~ 15：1, stirring liquid, dried object a are subjected to 1 ~ 3h of mixing, are heated to 38 ~ 50 DEG C, stand 25 ~
30min obtains mixed liquor, in mass ratio 10：4：0.1, mixed liquor, sodium hydroxide, extracting solution are mixed, get blisters 10 ~ 20min
Afterwards, 18 are freezed in temperature -20 ~ -10 DEG C ~ for 24 hours, then in 30 ~ 38 DEG C of 3 ~ 7d of heated-air drying of temperature, it is cut into small pieces, is sealed with paper wrapper
Sternly, it then in 100 ~ 120 DEG C of hot air sterilizations of temperature, is sealed in polybag to get the gelfoam for hemostasis.
Embodiment 1
Extract is that the rhizome of cogongrass is dried, and crushes, obtains crushed material, in mass ratio 1：7, by crushed material, mass fraction
It is mixed, the refluxing extraction under temperature 70 C, is filtered, separating filtrate, filter residue, 2 times repeatedly, merging filtrate for 80% ethyl alcohol,
Total filtrate is obtained, total filtrate is concentrated to get extract.
Crosslinking agent is glutaraldehyde.
Extracting solution is to be ground grape branch, obtains powder, in mass ratio 1：10, powder, 0.5mol/L ethyl alcohol is molten
Liquid is mixed, and in 40 DEG C of temperature, ultrasonic frequency carries out extraction 15min to get extracting solution under the conditions of being 100 Hz.
A kind of preparation method of gelfoam for hemostasis, the preparation method include the following steps：
（1）Jellyfish thaw broken, is placed in container, the papain of jellyfish quality 0.01% is added, is 5 in pH value,
It is hydrolyzed 1h under temperature 60 C, then the enzyme deactivation 8min in the water-bath of 98 DEG C of temperature, is filtered with double gauze, obtain filtrate；
（2）Filtrate is centrifuged into 5min with 800r/min, takes supernatant, be added into supernatant the ethyl alcohol of 1 times of supernatant quality into
Row alcohol precipitation 20h centrifuges 8min with 2000r/min, filters, filter cake is obtained, by filter cake ethyl alcohol, acetone alternating elution, in temperature 30
It is dry at DEG C, obtain dried object；
（3）Count in parts by weight, take 30 parts of distilled water, 10 parts of gelatin, 3 parts of extracting solutions, 1 part of crosslinking agent, by gelatin, distilled water into
Row mixing, impregnates 1h, then uniform in temperature 50 C stirred in water bath, adds dried object, crosslinking agent, stir evenly, must stir
Liquid, it is spare；
（4）In mass ratio 1：1：3, extract, polycaprolactone, dichloromethane are mixed, sonic oscillation 25min must vibrate
Liquid, in mass ratio 1：10, it will vibrate in liquid, 2% polyvinyl alcohol water solution of mass fraction, with 800r/min stirring 25min, then with
The closed stirring 2h of 100r/min centrifuge, obtain sediment, by precipitate washed with water, vacuum freezedrying obtains dried object a；
（5）In mass ratio 10：1, stirring liquid, dried object a are subjected to mixing 1h, 38 DEG C is heated to, stands 25min, obtain mixed liquor,
In mass ratio 10：4：0.1, mixed liquor, sodium hydroxide, extracting solution are mixed, after the 10min that gets blisters, in -20 DEG C of frosts of temperature
18h, then in 30 DEG C of heated-air drying 3d of temperature, be cut into small pieces, obturaged with paper wrapper, then in 100 DEG C of hot air sterilizations of temperature, be sealed in
To get the gelfoam for hemostasis in polybag.
Embodiment 2
Extract is that the rhizome of cogongrass is dried, and crushes, obtains crushed material, in mass ratio 1：8, by crushed material, mass fraction
It is mixed for 80% ethyl alcohol, the refluxing extraction at 80 DEG C of temperature, filtering, separating filtrate, filter residue, 3 times repeatedly, merging filtrate,
Total filtrate is obtained, total filtrate is concentrated to get extract.
Crosslinking agent is glutaraldehyde.
Extracting solution is to be ground grape branch, obtains powder, in mass ratio 1：10, powder, 0.5mol/L ethyl alcohol is molten
Liquid is mixed, and in temperature 60 C, ultrasonic frequency carries out extraction 20min to get extracting solution under the conditions of being 100 Hz.
A kind of preparation method of gelfoam for hemostasis, the preparation method include the following steps：
（1）Jellyfish thaw broken, is placed in container, the papain of jellyfish quality 0.02% is added, is 7 in pH value,
It is hydrolyzed 2h at 75 DEG C of temperature, then the enzyme deactivation 10min in the water-bath of 100 DEG C of temperature, is filtered with double gauze, obtain filtrate；
（2）Filtrate is centrifuged into 6min with 1000r/min, takes supernatant, be added into supernatant the ethyl alcohol of 2 times of supernatant quality into
For 24 hours, 3000r/min centrifuges 10min to row alcohol precipitation, filters, obtains filter cake, by filter cake ethyl alcohol, acetone alternating elution, in 40 DEG C of temperature
Lower drying, obtains dried object；
（3）Count in parts by weight, take 40 parts of distilled water, 15 parts of gelatin, 4 parts of extracting solutions, 2 parts of crosslinking agents, by gelatin, distilled water into
Row mixing, impregnates 2h, then uniform in 55 DEG C of stirred in water bath of temperature, adds dried object, crosslinking agent, stir evenly, must stir
Liquid, it is spare；
（4）In mass ratio 1：1：4, extract, polycaprolactone, dichloromethane are mixed, 25 ~ 30min of sonic oscillation must shake
Swing liquid, in mass ratio 1：15, by oscillation liquid, 3% polyvinyl alcohol water solution of mass fraction, 30min is stirred with 900r/min, then
With the closed stirring 3h of 200r/min, centrifuges, obtain sediment, by precipitate washed with water, vacuum freezedrying obtains dried object
a；
（5）In mass ratio 15：1, stirring liquid, dried object a are subjected to mixing 3h, 50 DEG C is heated, stands 30min, obtain mixed liquor, press
Mass ratio 10：4：0.1, mixed liquor, sodium hydroxide, extracting solution are mixed, after the 20min that gets blisters, in -10 DEG C of frosts of temperature
For 24 hours, then in 38 DEG C of heated-air drying 7d of temperature, it is cut into small pieces, is obturaged with paper wrapper, then in 120 DEG C of hot air sterilizations of temperature, is sealed in
To get the gelfoam for hemostasis in polybag.
Embodiment 3
Extract is that the rhizome of cogongrass is dried, and crushes, obtains crushed material, in mass ratio 1：7, by crushed material, mass fraction
It is mixed for 80% ethyl alcohol, the refluxing extraction at 75 DEG C of temperature, filtering, separating filtrate, filter residue, 3 times repeatedly, merging filtrate,
Total filtrate is obtained, total filtrate is concentrated to get extract.
Crosslinking agent is glutaraldehyde.
Extracting solution is to be ground grape branch, obtains powder, in mass ratio 1：10, powder, 0.5mol/L ethyl alcohol is molten
Liquid is mixed, and in temperature 50 C, ultrasonic frequency carries out extraction 18min to get extracting solution under the conditions of being 100 Hz.
A kind of preparation method of gelfoam for hemostasis, the preparation method include the following steps：
（1）Jellyfish thaw broken, is placed in container, the papain of jellyfish quality 0.01% is added, is 6 in pH value,
It is hydrolyzed 1.5h under temperature 70 C, then 8 ~ 10min of enzyme deactivation in the water-bath of 99 DEG C of temperature, is filtered, must be filtered with double gauze
Liquid；
（2）Filtrate is centrifuged into 5min with 900r/min, takes supernatant, be added into supernatant the ethyl alcohol of 1 times of supernatant quality into
Row alcohol precipitation 22h centrifuges 9min with 2500r/min, filters, filter cake is obtained, by filter cake ethyl alcohol, acetone alternating elution, in temperature 35
It is dry at DEG C, obtain dried object；
（3）Count in parts by weight, take 35 parts of distilled water, 13 parts of gelatin, 3 parts of extracting solutions, 1 part of crosslinking agent, by gelatin, distilled water into
Row mixing, impregnates 2h, then uniform in 53 DEG C of stirred in water bath of temperature, adds dried object, crosslinking agent, stir evenly, must stir
Liquid, it is spare；
（4）In mass ratio 1：1：3, extract, polycaprolactone, dichloromethane are mixed, 25 ~ 30min of sonic oscillation must shake
Swing liquid, in mass ratio 1：13, by oscillation liquid, 2% polyvinyl alcohol water solution of mass fraction, stirred with 800 ~ 900r/min
28min, then with the closed stirring 2h of 150r/min, centrifuge, obtain sediment, by precipitate washed with water, vacuum freezedrying,
Obtain dried object a；
（5）In mass ratio 10 ~ 15：1, stirring liquid, dried object a are subjected to mixing 2h, 42 DEG C is heated to, stands 28min, must mix
Liquid, in mass ratio 10：4：0.1, mixed liquor, sodium hydroxide, extracting solution are mixed, after the 15min that gets blisters, in -15 DEG C of temperature
20h is freezed, then in 33 DEG C of heated-air drying 4d of temperature, is cut into small pieces, is obturaged with paper wrapper, then in 110 DEG C of hot air sterilizations of temperature, it is close
It is encapsulated in polybag to get the gelfoam for hemostasis.
Comparative example：The gelfoam of Guangzhou company production.
It respectively selects 100 exceptions to hinder bleeding patients at random in hospital, is divided into experimental group and contrast groups, experimental group uses the present embodiment
Products obtained therefrom, contrast groups are stopped blooding using commercially available gelfoam.Application method：Gelfoam is taken directly to press on bleeding portion
Position.Curative effect situation criterion：Recovery from illness：Using 3 minutes, blood flow stopped completely, and pain is substantially reduced；Effectively：Using 3 minutes,
Blood flow is reduced, and pain is alleviated；In vain：Using 3 minutes, symptom was unchanged.Case infection conditions after observation hemostasis, standard
For：The apparent pain of surrounding tissue or redness, part pressure is well-illuminated aobvious, and laboratory leukocyte increases, and the results are shown in Table 1.
Table 1：
| Test event | Embodiment 1 | Embodiment 2 | Embodiment 3 | Comparative example |
| Recovery from illness（Example） | 82 | 80 | 84 | 27~30 |
| Effectively（Example） | 17 | 19 | 14 | 40~42 |
| In vain（Example） | 1 | 1 | 2 | 28~33 |
| Stop blooding postoperative infection（Example） | 0 | 1 | 0 | 6~10 |
In summary, not only haemostatic effect well more has anti-infection effect to gelfoam of the invention, is integrated compared to commercial product
Performance is more preferable, is worthy to be popularized.
Claims (4)
1. a kind of preparation method of gelfoam for hemostasis, which is characterized in that the preparation method includes the following steps：
（1）Jellyfish thaw broken, is placed in container, the papain of jellyfish quality 0.01 ~ 0.02% is added, in pH
Value is 5 ~ 7, is hydrolyzed at 60 ~ 75 DEG C of temperature, then the enzyme deactivation in the water-bath of 98 ~ 100 DEG C of temperature, is filtered, is obtained with double gauze
Filtrate；
（2）Filtrate is centrifuged, supernatant is taken, the ethyl alcohol that 1 ~ 2 times of supernatant quality is added into supernatant carries out alcohol precipitation, from
The heart filters, obtains filter cake, dry at 30 ~ 40 DEG C of temperature by filter cake ethyl alcohol, acetone alternating elution, obtains dried object；
（3）Count in parts by weight, take 30 ~ 40 parts of distilled water, 10 ~ 15 parts of gelatin, 5 ~ 6 parts of dried objects, 3 ~ 4 parts of extracting solutions, 1 ~ 2 part
Crosslinking agent mixes gelatin, distilled water, impregnates 1 ~ 2h, and extracting solution is added, uniform in 50 ~ 55 DEG C of stirred in water bath of temperature,
Dried object, crosslinking agent are added, is stirred evenly, stirring liquid is obtained, it is spare；
（4）In mass ratio 1：1：3 ~ 4, extract, polycaprolactone, dichloromethane are mixed, sonic oscillation, obtains oscillation liquid,
In mass ratio 1：10 ~ 15, by oscillation liquid, 2 ~ 3% polyvinyl alcohol water solution of mass fraction, stirring, then closed stirring, centrifugation divide
From obtaining sediment, by precipitate washed with water, vacuum freezedrying obtains dried object a；
（5）In mass ratio 10 ~ 15：1, stirring liquid, dried object a are mixed, are heated to 38 ~ 50 DEG C, stands, obtains mixed liquor,
In mass ratio 10：4：0.1, mixed liquor, sodium hydroxide, extracting solution are mixed, got blisters, is freezed in temperature -20 ~ -10 DEG C, then
In 30 ~ 38 DEG C of heated-air dryings of temperature, it is cut into small pieces, is obturaged with paper wrapper, then in 100 ~ 120 DEG C of hot air sterilizations of temperature, is sealed in
To get the gelfoam for hemostasis in polybag.
2. being used for the preparation method of the gelfoam of hemostasis according to claim 1, which is characterized in that the step（3）In
Extracting solution is that the rhizome of cogongrass is dried, and crushes, obtains crushed material, in mass ratio 1：7 ~ 8, be by crushed material, mass fraction
80% ethyl alcohol is mixed, the refluxing extraction at 70 ~ 80 DEG C of temperature, filtering, separating filtrate, filter residue, 2 ~ 3 times repeatedly, merges filter
Liquid is to get extracting solution.
3. being used for the preparation method of the gelfoam of hemostasis according to claim 1, which is characterized in that the step（3）In
Crosslinking agent is glutaraldehyde.
4. being used for the preparation method of the gelfoam of hemostasis according to claim 1, which is characterized in that the step（5）In
Extracting solution is to be ground grape branch, obtains powder, in mass ratio 1：10, powder, 0.5mol/L ethanol solutions are mixed
It closes, in 40 ~ 60 DEG C of temperature, ultrasonic frequency carries out 15 ~ 20min of extraction to get extracting solution under the conditions of being 100 Hz.
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