CN109769808A - A kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent and preparation method - Google Patents
A kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent and preparation method Download PDFInfo
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Abstract
The invention discloses a kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent, chitosan is reacted as carrier material by emulsification and cross linked with gelatin is wherein made sustained-release microparticle for the embedding of phenolic acid class Acting mode；Preparation step: phenolic acid class Acting mode is added in the glacial acetic acid solution containing chitosan, and the atoleine high temperature containing span80 is added in this solution and is emulsified uniformly；Aqueous gelatin solution is added in above-mentioned emulsion again, stirring and emulsifying, glutaraldehyde cross-linking solidification is added dropwise after cooling；Centrifugation, washing precipitate, and are dried in vacuo after suction filtration and sustained-release microparticle is made.The present invention is by phenolic acid class Acting mode in conjunction with micro-nano capsule technique, it is small that partial size is made, it is not easy the sustained-release microparticle algae-inhibiting agent that expansion is sunk, extend phenolic acid class Acting mode in water body to the inhibition time of microcystic aeruginosa, reduce cost, applied to true water body algal control field, new technological approaches is provided for natural water algal control for phenolic acid class algistatic activity substance.
Description
Technical field
The present invention relates to a kind of algae-inhibiting agent and preparation methods, and in particular to a kind of load medicine chitosan/gelatin sustained-release microparticle suppression
Algae agent and preparation method belong to water eutrophication and inhibit alga eruption technical field.
Background technique
Microcapsules are controlled because of its control release performance excellent with long-acting, efficient, targeting, low side effect etc. in drug
The fields such as release have broad application prospects.Since this technology is grown up in cross discipline, at home and abroad
Through the research hotspot for becoming the multidisciplinary field workers such as material, chemistry, chemical industry, biology and medicine.Drug-carried fine particle sustained release is one
Kind controls the rate of release for the drug that is embedded and the technology of release direction, and research application is less in terms of the improvement of Ecology.
Sustained-release microparticle carrier material generally can be divided into natural macromolecular material and synthesis high molecular material；Wherein, natural high
Molecular material because its is nontoxic, stable, biodegradable and biocompatibility preferably etc. characteristics in research in recent years using compared with
It is more.
Gelatin is the denatured protein extracted by acid or basic hydrolysis animal collagen, is that amino acid and peptides are crosslinked
The straight chain polymer compound of formation, because its is cheap, it is easy obtain, nontoxic, water-soluble, good biocompatibility and can biology
The characteristics such as degradation and be especially widely used in terms of medicament slow release and release in terms of the biomedical material.
Acting mode algal control has the characteristics that efficient, quick and ecological security is good.
The present invention combines Acting mode algal control with micro-nano capsule technique, provides for true water body algal control new
Technological approaches.
Summary of the invention
To solve the deficiencies in the prior art, that the purpose of the present invention is to provide a kind of partial sizes is small, is not easy expansion sinks, is nontoxic
Load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent.
In order to achieve the above objectives, the present invention adopts the following technical scheme that:
A kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent, including sustained-release microparticle and embedding phenolic acid class allelopathic activity therein
Substance；
The sustained-release microparticle is formed as carrier material by emulsification and cross linked reaction bonded by chitosan with gelatin.
A kind of above-mentioned load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent preparation method, comprising the following steps:
S1, chitosan is dissolved in glacial acetic acid solution, chitosan solution is made, while it is added in phenolic acid class Acting mode
In, it is made and carries medicine chitosan solution；
S2, it will carry in atoleine of the medicine chitosan solution addition containing span80, emulsion is made in stirring and emulsifying；
S3, aqueous gelatin solution is instilled in above-mentioned emulsion, continues stirring and emulsifying, it is cooling；
After S4, cooling, glutaraldehyde is added, stirring solidifies, is centrifuged, and washs, filters sediment, is dried in vacuo obtained load medicine shell and gathers
Sugar/gelatin sustained-release microparticle.
The mass ratio of chitosan and phenolic acid class Acting mode in above-mentioned steps S1 is 10:1 ~ 1:1.
Atoleine and water phase volume ratio are 2:1 ~ 8:1 in above-mentioned steps S2, span80 mass content be oily phase 1% ~
4%。
Gelatin/chitosan mass ratio in above-mentioned steps S3 is 1:1 ~ 4:1.
Glutaraldehyde content in above-mentioned steps S4 is the 1% ~ 4% of system solution volume.
The mass concentration of glacial acetic acid solution in above-mentioned steps S1 is 1% ~ 3%, the concentration of chitosan solution be 5mg/ml ~
15mg/ml。
Stirring in above-mentioned steps S2 includes magnetic agitation, and speed of agitator is 800r/min ~ 1000r/min, whipping temp
It is 45 DEG C ~ 55 DEG C, emulsification times are 10min ~ 20min.
Gelatin in above-mentioned steps S3 is dissolved in 50 DEG C ~ 65 DEG C of aqueous solution, the concentration of aqueous gelatin solution be 10mg/ml ~
30mg/ml；Stirring includes magnetic agitation, and speed of agitator is 800r/min ~ 1000r/min, and whipping temp is 45 DEG C ~ 55 DEG C, cream
The change time is 10min ~ 20min；Cooling temperature is 4 DEG C ~ 15 DEG C.
It is 30min ~ 90min that curing time is stirred in above-mentioned steps S4；The revolving speed of centrifugation is 600r/min ~ 1000r/min,
Centrifugation time is 10min ~ 15min；Detergent includes petroleum ether and dehydrated alcohol, and washing times at least 3 times；Dry environment be
It is dried in vacuo at 35 DEG C ~ 45 DEG C.
Above-mentioned phenolic acid class Acting mode includes: gallic acid, pyrogallic acid, ferulic acid, catechol.
The invention has the beneficial effects that:
A kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent of the invention and preparation method, by phenolic acid class Acting mode
In conjunction with micro-nano capsule technique, by using chitosan and gelatin to embed phenolic acid class Acting mode, system as carrier material
At sustained-release microparticle algae-inhibiting agent, particle size is 40 μm ~ 80 μm, is not easy expansion and sinks, can extend phenolic acid class Acting mode
In water body to the inhibition time of microcystic aeruginosa, reduce cost, efficiently solve that directly to add phenolic acid class allelopathic to water body living
Property substance and the active material dosage that generates is big, be easy to run off, part phenolic acid is insoluble in water, algal control concentration etc. is not achieved
Problem and the disadvantages of the resulting slow-releasing granules partial size of early-stage study is larger, easy sinking.
Chitosan used in the present invention, gelatin are natural polymer nontoxic, that biocompatibility is preferable, biodegradable
Material can effectively avoid the secondary pollution during algal control, be applied to true water body algal control field, can be widely applied to pond,
The natural waters such as reservoir, lake inhibit the outburst of algae, provide for phenolic acid class algistatic activity substance for natural water algal control
New technological approaches.
Detailed description of the invention
Fig. 1 is inhibition figure of the embodiment of the present invention 1 to microcystic aeruginosa.
Specific embodiment
Below in conjunction with detailed description of the invention by specific embodiments.
Reagent used in the present invention is commercially available.
Phenolic acid class Acting mode selects gallic acid in the embodiment of the present invention.
Embodiment 1:
S1, it weighs 0.1g chitosan and is dissolved in 1% glacial acetic acid solution of 10ml, while by 0.02g phenolic acid class Acting mode
It is added thereto；
S2, will carry medicine chitosan solution be added the span80 containing 3.2ml 80ml atoleine in, with 1000r/ at 50 DEG C
The revolving speed of min stirs evenly, and emulsifies 15min；
S3, then the aqueous gelatin solution of the 20mg/ml of 10ml instilled in above-mentioned emulsion, continues stirring and emulsifying at 50 DEG C
15min is subsequently agitated for being cooled to 15 DEG C；
S4,1.5ml glutaraldehyde, stirring solidification 60min is added；10min is centrifuged under conditions of 800r/min, precipitating uses petroleum ether
It is filtered after being washed 3 times with dehydrated alcohol, is finally dried in vacuo at 40 DEG C and load medicine chitosan/gelatin sustained-release microparticle is made.
Acute toxicity reality is carried out to the microcystic aeruginosa of sterile culture using medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent is carried
It tests:
Three parallel groups are set, and observation is primary for 24 hours, is observed continuously one week, carries medicine chitosan/gelatin sustained-release microparticle algal control when being added
When agent concentration is 1.2g/L, 72% is reached to the 96h inhibiting rate of microcystic aeruginosa, is observed continuously 30 days, microcystic aeruginosa inhibits
Rate reaches 99%.
Embodiment 2:
S1, it weighs 0.2g chitosan and is dissolved in 1% glacial acetic acid solution of 20ml, while by 0.04g phenolic acid class Acting mode
It is added thereto；
S2, it will carry in the atoleine of 150ml of the medicine chitosan solution addition containing 6mlspan80, with 1000r/min at 50 DEG C
Revolving speed stir evenly, emulsify 15min；
S3, then the aqueous gelatin solution of the 20mg/ml of 30ml instilled in above-mentioned emulsion, continues stirring and emulsifying at 50 DEG C
15min is subsequently agitated for being cooled to 15 DEG C；
S4,2.5ml glutaraldehyde, stirring solidification 60min is added；10min is centrifuged under conditions of 800r/min, precipitating uses petroleum ether
It is filtered after being washed 3 times with dehydrated alcohol, is finally dried in vacuo at 40 DEG C and load medicine chitosan/gelatin sustained-release microparticle is made.
Acute toxicity reality is carried out to the microcystic aeruginosa of sterile culture using medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent is carried
It tests:
Three parallel groups are set, and observation is primary for 24 hours, is observed continuously one week, carries medicine chitosan/gelatin sustained-release microparticle algal control when being added
When agent concentration is 1.2g/L, 70% is reached to the 96h inhibiting rate of microcystic aeruginosa, is observed continuously 30 days, microcystic aeruginosa inhibits
Rate reaches 95%.
Embodiment 3
S1, it weighs 0.15g chitosan and is dissolved in 1% glacial acetic acid solution of 15ml, while by 0.03g phenolic acid class allelopathic active matter
Matter is added thereto；
S2, it will carry in the atoleine of 100ml of the medicine chitosan solution addition containing 3.5mlspan80, with 800r/ at 50 DEG C
The revolving speed of min stirs evenly, and emulsifies 15min；
S3, then the aqueous gelatin solution of the 20mg/ml of 20ml instilled in above-mentioned emulsion, continues stirring and emulsifying at 50 DEG C
15min is subsequently agitated for being cooled to 15 DEG C；
S4,2ml glutaraldehyde, stirring solidification 60min is added；Be centrifuged 10min under conditions of 800r/min, precipitating petroleum ether and
Dehydrated alcohol filters after washing 3 times, is finally dried in vacuo at 40 DEG C and load medicine chitosan/gelatin sustained-release microparticle is made.
Acute toxicity reality is carried out to the microcystic aeruginosa of sterile culture using medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent is carried
It tests:
Three parallel groups are set, and observation is primary for 24 hours, is observed continuously one week, carries medicine chitosan/gelatin sustained-release microparticle algal control when being added
When agent concentration is 1.2g/L, 60% is reached to the 96h inhibiting rate of microcystic aeruginosa, is observed continuously 30 days, microcystic aeruginosa inhibits
Rate reaches 94%.
Embodiment 4
S1, it weighs 0.2g chitosan and is dissolved in 1% glacial acetic acid solution of 20ml, while by 0.03g phenolic acid class Acting mode
It is added thereto；
S2, it will carry in the atoleine of 160ml of the medicine chitosan solution addition containing 6.5mlspan80, with 900r/ at 50 DEG C
The revolving speed of min stirs evenly, and emulsifies 15min；
S3, then the aqueous gelatin solution of the 20mg/ml of 30ml instilled in above-mentioned emulsion, continues stirring and emulsifying at 50 DEG C
15min is subsequently agitated for being cooled to 15 DEG C；
S4,3ml glutaraldehyde, stirring solidification 60min is added；Be centrifuged 10min under conditions of 800r/min, precipitating petroleum ether and
Dehydrated alcohol filters after washing 3 times, is finally dried in vacuo at 40 DEG C and load medicine chitosan/gelatin sustained-release microparticle is made.
Acute toxicity reality is carried out to the microcystic aeruginosa of sterile culture using medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent is carried
It tests:
Three parallel groups are set, and observation is primary for 24 hours, is observed continuously one week, carries medicine chitosan/gelatin sustained-release microparticle algal control when being added
When agent concentration is 1.2g/L, 63% is reached to the 96h inhibiting rate of microcystic aeruginosa, is observed continuously 30 days, microcystic aeruginosa inhibits
Rate reaches 93%.
By comparing the chlorophyll-a concentration of example 1 and blank microcystic aeruginosa liquid, such as Fig. 1.It can significantly find, it is real
The chlorophyll-a concentration that load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent microcystic aeruginosa is added in example 1 is first increased and is reduced afterwards,
Algae is in abnormal growth state later, and inhibitory effect is obvious.Chitosan and gelatin after glutaraldehyde cross-linking by combining more
Closely, algae-inhibiting agent stability is good, and active material stablizes release in sustained-release microparticle in experimentation, and chlorophyll-a concentration was at the 10th day
After start to reduce, be constantly in lower concentration later.
The basic principles, main features and advantages of the invention have been shown and described above.The technical staff of the industry should
Understand, the above embodiments do not limit the invention in any form, all obtained by the way of equivalent substitution or equivalent transformation
Technical solution is fallen within the scope of protection of the present invention.
Claims (10)
1. a kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent, which is characterized in that including sustained-release microparticle and embed phenol therein
Acids Acting mode；
The sustained-release microparticle is formed as carrier material by emulsification and cross linked reaction bonded by chitosan with gelatin.
2. a kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent preparation method according to claim 1, feature exist
In, comprising the following steps:
S1, chitosan is dissolved in glacial acetic acid solution, chitosan solution is made, while it is added in phenolic acid class Acting mode
In, it is made and carries medicine chitosan solution；
S2, it will carry in atoleine of the medicine chitosan solution addition containing span80, emulsion is made in stirring and emulsifying；
S3, aqueous gelatin solution is instilled in above-mentioned emulsion, continues stirring and emulsifying, it is cooling；
After S4, cooling, glutaraldehyde is added, stirring solidifies, is centrifuged, and washs, filters sediment, is dried in vacuo obtained load medicine shell and gathers
Sugar/gelatin sustained-release microparticle.
3. a kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent preparation method according to claim 2, feature exist
In the mass ratio of chitosan and phenolic acid class Acting mode in the step S1 is 10:1 ~ 1:1.
4. a kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent preparation method according to claim 2, feature exist
In atoleine and water phase volume ratio are 2:1 ~ 8:1 in the step S2, and span80 mass content is the 1% ~ 4% of oily phase.
5. a kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent preparation method according to claim 2, feature exist
In the gelatin/chitosan mass ratio in the step S3 is 1:1 ~ 4:1.
6. a kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent preparation method according to claim 2, feature exist
In the glutaraldehyde content in the step S4 is the 1% ~ 4% of system solution volume.
7. a kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent preparation method according to claim 2, feature exist
In the mass concentration of the glacial acetic acid solution in the step S1 is 1% ~ 3%, and the concentration of chitosan solution is 5mg/ml ~ 15mg/
ml。
8. a kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent preparation method according to claim 2, feature exist
In the stirring in the step S2 includes magnetic agitation, and speed of agitator is 800r/min ~ 1000r/min, and whipping temp is 45 DEG C
~ 55 DEG C, emulsification times are 10min ~ 20min.
9. a kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent preparation method according to claim 2, feature exist
In the gelatin in the step S3 is dissolved in 50 DEG C ~ 65 DEG C of aqueous solution, and the concentration of aqueous gelatin solution is 10mg/ml ~ 30mg/
ml；Stirring includes magnetic agitation, and speed of agitator is 800r/min ~ 1000r/min, and whipping temp is 45 DEG C ~ 55 DEG C, emulsification times
For 10min ~ 20min；Cooling temperature is 4 DEG C ~ 15 DEG C.
10. a kind of load medicine chitosan/gelatin sustained-release microparticle algae-inhibiting agent preparation method according to claim 2, feature
It is, it is 30min ~ 90min that curing time is stirred in the step S4；The revolving speed of centrifugation is 600r/min ~ 1000r/min, from
The heart time is 10min ~ 15min；Detergent includes petroleum ether and dehydrated alcohol, and washing times at least 3 times；Dry environment is 35
DEG C ~ 45 DEG C at be dried in vacuo.
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