CN107789332A - A kind of calcium carbonate/calcium alginate compounded microballoon that adjustable drug release rate is prepared based on aqueous two-phase biomineralization technology - Google Patents
A kind of calcium carbonate/calcium alginate compounded microballoon that adjustable drug release rate is prepared based on aqueous two-phase biomineralization technology Download PDFInfo
- Publication number
- CN107789332A CN107789332A CN201710768009.0A CN201710768009A CN107789332A CN 107789332 A CN107789332 A CN 107789332A CN 201710768009 A CN201710768009 A CN 201710768009A CN 107789332 A CN107789332 A CN 107789332A
- Authority
- CN
- China
- Prior art keywords
- solution
- calcium
- calcium carbonate
- calcium alginate
- phase
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/20—Pills, tablets, discs, rods
- A61K9/28—Dragees; Coated pills or tablets, e.g. with film or compression coating
- A61K9/2886—Dragees; Coated pills or tablets, e.g. with film or compression coating having two or more different drug-free coatings; Tablets of the type inert core-drug layer-inactive layer
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/38—Albumins
- A61K38/385—Serum albumin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/02—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
- B01J13/02—Making microcapsules or microballoons
- B01J13/06—Making microcapsules or microballoons by phase separation
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Inorganic Chemistry (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Dispersion Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention discloses a kind of calcium carbonate/calcium alginate compounded microballoon that adjustable drug release rate is prepared based on aqueous two-phase biomineralization technology and preparation method thereof, belongs to high polymer material and controlled release drug carrier field.Preparation method of the present invention, simple to operate, power consumption is low, and system moderate notoxic；Selective partition characteristic using aqueous two-phase to macro-molecular protein, macro-molecular protein class medicine can be retained in well in the interior phase of aqueous two-phase drop template, the inside of compound calcium alginate microballoon is admirably encapsulated in solidification, there is excellent envelop rate to protein drug；And calcium carbonate/calcium alginate microsphere of different composite degree by the urase concentration of regulation system, can be prepared, and then the rate of release of controllable embedding medicine therein；It is larger to overcome conventional composite calcium alginate microsphere network hole, contains the shortcomings that thing easily flows out.Can widely develop its fields such as biologic medical, cosmetic industry, food service industry and enzyme engineering application.
Description
Technical field
The present invention relates to polymeric material field, especially a kind of carbonic acid prepared using aqueous two-phase biomineralization technology
Calcium/calcium alginate compounded microballoon and preparation method thereof.
Aqueous two-phase system is the aqueous solution or a kind of high molecular polymer aqueous solution and one by two kinds of high molecular polymers
Kind salting liquid is formed.When two kinds of solution concentrations exceed some threshold value, two kinds of solution of mixing can spontaneous generation split-phase, shape
All it is the two-phase system of the aqueous solution, as aqueous two-phase system into about one two-phase.Aqueous two-phase system has to bioactive substance
There is the advantages of high biocompatibility, while there is steric effect, macromolecular substances are not easy face transmission transboundary, so double water
Mutually there is extensive use in terms of biology.Cacace etc. is using two-phase polymer aqueous medium simulation macromolecular crowding and carefully
Intracellular environment divides, and carries out biomineralization in polyethylene glycol and dextran double-aqueous phase system, that is, utilizes urase catalyzing urea
CO caused by hydrolysis3 2-Be present in solution calcium ion reaction production calcium carbonate ([1]David N.Cacace,
Chrisstine D.keating,Biocatalyzed mineralization in an aqueous two-phase
system:effect of background polymers and enzyme partitioning[J].Journal
Material Chemistry B,2013,1,1794-1803)。
Biomineralization refers to organism and adjusted by itself to control the generating process of inorganic mineral, is that one kind can be prepared again
The straightforward procedure of miscellaneous structure, got well much than artificial synthesized by the combination property of this method resulting materials.Xie etc. utilizes sea
Alginates control to form the crystallization process of nanometer grade calcium carbonate and hydroxyapatite, and its achievement in research is advantageous to composite wood from now on
Material design ([2]M. Xie,M.J.P.Andreassen,S.M.Selbach,B.L.Strand,P.Sikorski
Alginate-controlled formation of nanoscale calcium carbonate and
hydroxyapatite mineral phase within hydrogel networks[J].Acta Biomaterialia,
2010,6(9):3665-3675)。
Calcium alginate microsphere Nantural non-toxic, has good a biocompatibility and biodegradability, preparation process is simple,
Gently, safety, is now widely used for biological respinse engineering and medical transmission systems.Such as Moebus is high by being spray-dried
Molecular medicine solution prepares the calcium alginate microsphere of load bovine serum albumin(BSA) (BSA), but spray drying device high energy consumption, system
Standby process it is complicated ([3]Moebus K,Siepmann J,Bodmeier R.Novel preparation techniques for
alginate-poloxamer microparticles controlling protein release on mucosal
surfaces.European Journal of Pharmaceutical Sciences,2012,45(3):358-366)。
However, single calcium alginate hydrogel microballoon has, network pore-size is larger, and mechanical strength is poor, hygroscopic
Crushed after swelling, cause to contain logistics and, the shortcomings that repeat usage is not high.Song Limin etc. prepares chitosan using emulsion process
The calcium alginate compounded microballoon of overlay film, the mechanical strength of calcium alginate is improved, but preparation process is unavoidably using toxic
Organic reagent, and can not keep biopharmaceutical macromolecular drug activity ([4]Song Limin, Li Ming, Ni Yaxin, Ni Tian, Gao Mengmeng,
Preparation [J] application chemical industry of Wang Xiaolei, Zhang Baohua effective cypermethrins/alginate/chitosan gel micro-ball, 2016,
(11):2118-2124.)。
Shi et al. uses aliphatic polyurethane-amine (PU) to be used as thermal response component, in high pressure CO2Under the conditions of, use is bionical
Mineralising technology prepares alginates/calcium carbonate hybrid pearl with pH and thermal response drug release characteristics, and uses polyacrylic acid
(PAA) as crystal growth additive come control the structure of hydridization pearl ([5]Shi,J.；Zhang,Z.；Li,G.；Cao,
S.Biomimetic Fabrication of Alginate/Caco3Hybrid Beads for Dual-Responsive
Drug Delivery under Compressed CO2[J].Journal Material Chemistry B,2011,21
(40)16028-16034.)
This kind of method reaction condition is violent, is equally also unfavorable for biopharmaceutical macromolecular drug and keeps activity.Isiklan etc. is utilized
Glutaraldehyde makees crosslinking agent, and hydrochloric acid is made the copolymer material of the alginate of catalyst preparation nifepine and Itaconic Acid Grafted, is used for
Insoluble drug release ([6]Isiklan.N,Inal.M,Kursun.F,et al,pH responsive itaconic acid
grafted alginate microspheres for the controlled release of nifedipine[J]
.Carbohydrate Polymers,2011,84(3):933).
The inevitable use of this kind of method is crosslinked to the virose glutaraldehyde of human body.Meanwhile calcium alginate microsphere because
Its gel network is larger, and the envelop rate of entrapped drug is not high always, causes the waste of embedded object such as medicine and enzyme.More than being based on,
Inventor overcomes the shortcomings of that tradition prepares complex microsphere method, while the shortcomings that improve single calcium alginate microsphere, devises
A kind of calcium carbonate/calcium alginate compounded microballoon prepared using aqueous two-phase system by biomineralization technology.The complex microsphere exists
Aqueous two-phase system benign environment and without using other poisonous and harmful reagents under conditions of, using aqueous two-phase to macro-molecular protein
Selective partition characteristic, macro-molecular protein class medicine can be retained in well in the interior phase of aqueous two-phase drop template, entered
And the inside of compound calcium alginate microballoon is admirably encapsulated in solidification, there is excellent envelop rate, reduce raw material
Loss., can be to change calcium carbonate/calcium alginate and the compound calcium alginate is by regulation system response parameter-enzyme concentration
The compactness extent of the calcium carbonate of microballoon, and then can flexible modulation its internal drug release rate.Compared with other method, we
Method avoids other method in outer layer coated high molecular material, more easy.The method that the complex microsphere utilizes biomimetic mineralization,
Calcium carbonate crystal is grown among calcium alginate microsphere gel network, overcome single calcium alginate hydrogel microballoon net
Network hole is larger, contains the shortcomings that thing easily flows out.Can widely develop its biologic medical, cosmetic industry, food service industry and
The application in the fields such as enzyme engineering.
The content of the invention
The purpose of the present invention is to prepare a kind of calcium carbonate/calcium alginate compounded micro- by aqueous two-phase biomineralization technology
Ball, the microballoon has excellent envelop rate to protein medicaments, while can be realized by changing enzymatic reaction parameter-enzyme concentration
Allow loading medicine to possess different release rates, there is more preferable regulation performance compared to single calcium alginate microsphere, so as to apply
In industrial structure step slow release medicine system.It is a further object to provide a kind of aqueous two-phase biomineralization skill
Art prepares the preparation method of calcium carbonate/calcium alginate compounded microballoon.
Realize that the technical scheme of the object of the invention is as follows：
A kind of calcium carbonate/calcium alginate that adjustable drug release rate is prepared based on PEG/Dex aqueous two-phase biomineralization technologies
Complex microsphere, calcium carbonate/calcium alginate compounded microballoon are shell solid construction, and its shell is that shape is blended with calcium carbonate in calcium alginate
Into compound, its layer is that the calcium carbonate that mineralising reaction is formed is grown on grid in the layer of calcium alginate microsphere, and can be encapsulated big
The protein drug of molecule；Complex microsphere particle diameter distribution is 200~600 μm.
Further, the selective partition characteristic using the aqueous two-phase to macro-molecular protein, by the albumen of macromolecular
Interior phase of the matter class drug encapsulation in the Dex aqueous two-phases drop template.
Further, the Dex drops are that template forms microballoon using drop template；And then cause macro-molecular protein
Class medicine is encapsulated in compound calcium alginate microballoon when solidifying and adjusting.
Further, by adjusting different enzyme reaction condition-urase concentration, can to prepare different calcium carbonate/calcium alginates multiple
The microballoon of conjunction degree, and obtain different drug release rates.
Further, the aqueous two-phase drop template is that a certain amount of urea and a certain amount of calcium chloride are scattered in into 8%
In polyethylene glycol phase PEG (molecular weight 8kDa), a certain amount of urase and a certain amount of sodium alginate are scattered in 8% glucan phase Dex
In (molecular weight 500kDa)；Dex phases are instilled in PEG phases by the tip capillary that tip internal diameter is 20-80 μm and form carbonic acid
Calcium/calcium alginate compounded microballoon.
A kind of calcium carbonate/calcium alginate that adjustable drug release rate is prepared based on PEG/Dex aqueous two-phase biomineralization technologies
The preparation method of complex microsphere, it is comprised the following steps that：
First step：The preparation of PEG-Dex aqueous two-phases：
(1) mass fraction 10-20% PEG solution, molecular weight 8kDa are configured to；
(2) mass fraction 10-20% Dex solution, molecular weight 500kDa are configured to；
(3) take step (1) and step (2) solution to be sufficiently mixed, stand split-phase, obtain phase and be rich in PEG, lower phase is rich in
Dex aqueous two-phase solution；Extract PEG solution respectively and Dex solution is standby into two beakers；
Second step：Drop template prepares calcium carbonate/calcium alginate compounded microballoon：
(1) solution A：0.01-0.10g sodium alginates and 0.002-0.050g urases are weighed, is added to the above-mentioned of 5-10mL
In Dex solution, it is well mixed, set aside for use；
(2) B solution：Anhydrous calcium chloride 0.1-0.5g and urea 0.1-0.5g are dissolved in 5-10mL PEG solution,
It is well mixed, set aside for use；
(3) solution A is sucked in common commercially available syringe, by tip internal diameter is 20-80 μm with commercial silica gel pipe, its pipe shaft
Internal diameter is that 500 μm of capillary is connected with commercially available syringe, and solution A is advanced in tip capillary by syringe, and will
Solution A is vertically added dropwise in B solution by the way that tip capillary is hanging with the speed of 2 seconds one drops, and its capillary tip is apart from liquid level
0.5-5cm, a diameter of 200~600 μm of calcium alginate microsphere can be formed after the several seconds；Continue to place 1-10 at 20-60 DEG C
After h, treat that enzymatic reaction is fully carried out, produce calcium carbonate/calcium alginate compounded microballoon of adjustable drug release rate.
The present invention compared with prior art, has the advantages that：
1st, calcium carbonate of the present invention/calcium alginate compounded microballoon introduces double-aqueous phase system, is divided greatly using aqueous two-phase
Sub- steric effect and specific selectivity so that protein substance of the complex microsphere for macromolecular made from drop template
With high envelop rate, for example, measure BSA envelop rate be 97.27%.
2nd, complex microsphere of the present invention had both had the advantages that the biocompatibility of calcium alginate, bioadhesive, led to
Prepared by the technology for crossing biomineralization, the advantages of making it have bionical mineral high mechanical strength, make it be not easy to be swelled.
3rd, in preparation process, regulation system enzyme concentration is passed through, thus it is possible to vary the shell of calcium carbonate/calcium alginate microsphere is fine and close
Degree, and then its rate of release is adjusted, realization allows loading medicine to possess different release rates.
4th, calcium carbonate of the present invention/calcium alginate compounded microballoon is nontoxic using reagent, and preparation process condition
Gently.Had broad application prospects in fields such as biologic medical, cosmetic industry, food service industry and chemical materials；Especially
It can be applied to build step slow release medicine system.
5th, it is of the invention compared with other methods, other methods are avoided in outer layer coated high molecular material, it is more easy.Profit
With the method for biomimetic mineralization, calcium carbonate crystal is grown among calcium alginate microsphere gel network, overcome single marine alga
Sour calcium hydrogel microsphere fenestral porosity is larger, contains the shortcomings that thing easily flows out.
Brief description of the drawings
Fig. 1 is the schematic diagram that drop template of the present invention prepares calcium carbonate/calcium alginate microsphere into method.
Fig. 2A 1 is that urase concentration is 5mgmL-1When obtained calcium carbonate/calcium alginate microsphere microphotograph signal
Figure.
Fig. 2A 2 is Fig. 2A 1 micro- schematic diagram of partial enlargement.
Fig. 2 B are that urase concentration is 5mgmL-1When obtained calcium carbonate/calcium alginate microsphere complex microsphere in encapsulate
FITC-BSA fluorescence photo schematic diagram.
Fig. 2 C figures are that urase concentration is respectively 0-5mgmL-1When obtained calcium carbonate/calcium alginate microsphere digital photograph
Piece schematic diagram.
Fig. 3 A are that common alginic acid calcisphere embedding small molecule dimethyl diaminophenazine chloride of the present invention is released in 0min, 10min, 20min
Put design sketch (being mixed without calcium carbonate).
Fig. 3 B are that the compound calcium alginate ball of urase reaction generation embeds small molecule dimethyl diaminophenazine chloride in 0min, 10min, 20min
Releasing effect figure (has calcium carbonate mixing).
Fig. 3 C be ammonium carbonate/calcium chloride directly react formation compound calcium alginate microballoon embedding small molecule dimethyl diaminophenazine chloride exist
0min, 10min, 20min releasing effect figure (fine and close calcium carbonate mixing).
Fig. 4 is the bag of calcium carbonate/calcium alginate microsphere encapsulating various concentrations bovine serum albumin BSA of preparation of the present invention
The column schematic diagram of envelope rate.
Fig. 5 is that calcium carbonate/calcium alginate microsphere prepared by different enzyme concentrations of the present invention contains bovine serum albumin BSA and released
Put the column schematic diagram of rate.
Part instrument：Instrument biomicroscope, model：XSP-24, manufacturer：Phoenix Optics Holding Co., Ltd., production
Ground：Jiangxi, China Shangrao.Canon's camera, model：SX260HS, manufacturer：CANON (CHINA) Co Ltd, the place of production：China
Beijing.
Embodiment
The present invention is further illustrated with reference to drawings and Examples.
Fig. 1 describes the mechanism and example that aqueous two-phase drop template prepares calcium carbonate/calcium alginate microsphere.The Dex
Drop is that template forms microballoon using drop template；And then macro-molecular protein class medicine is encapsulated in solidification
In compound calcium alginate microballoon.Even if a certain amount of urea and a certain amount of calcium chloride are scattered in 8% polyethylene glycol phase PEG (molecular weight
In 8kDa), a certain amount of urase and a certain amount of sodium alginate are scattered in 8% glucan phase Dex (molecular weight 500kDa)；Pass through
Dex phases are instilled formation calcium carbonate/calcium alginate compounded microballoon in PEG phases by the tip capillary that tip internal diameter is 20-80 μm.
Can be seen that from Fig. 2A 1, Fig. 2A 2, Fig. 2 B, Fig. 2 C prepared based on PEG/Dex aqueous two-phase biomineralization technologies it is adjustable
The calcium carbonate of drug release rate/calcium alginate compounded microballoon, is shell solid construction, and its shell is that calcium alginate is blended with calcium carbonate
The compound of formation, its layer is that the calcium carbonate that mineralising reaction is formed is grown on grid in the layer of calcium alginate microsphere, and can be encapsulated
The protein drug of macromolecular.Complex microsphere particle diameter distribution is 200~600 μm.Using aqueous two-phase of the present invention to big point
The selective partition characteristic of sub- protein, the protein drug of macromolecular is encapsulated in the Dex aqueous two-phases drop template
Interior phase.The figures of Fig. 2A 1 are that urase concentration is 5mgmL-1When obtained calcium carbonate/calcium alginate microsphere microphotograph, figure
2A2 is Fig. 2A 1 partial enlargement micrograph, and black particle is calcium carbonate granule in figure, and hyalomere is divided into calcium alginate gel.
Fig. 3 B are that urase concentration is 5mgmL-1When obtained calcium carbonate/calcium alginate microsphere complex microsphere in the FITC-BSA that encapsulates
Fluorescence photo.What the green portion of mark represented is FITC-BSA position.Fig. 3 C are that urase concentration is respectively 0-
5mg·mL-1When obtained calcium carbonate/calcium alginate microsphere exterior appearance digital photograph.As a result obtained by the explanation present invention
Complex microsphere is that calcium carbonate is blended together with calcium alginate, and for the BSA overwhelming majority in complex microsphere at interface
And microballoon internal layer, good containment is made it have, Fig. 3 A are that common alginic acid calcisphere of the present invention mixes without calcium carbonate
Small molecule dimethyl diaminophenazine chloride is embedded during conjunction in 0min, 10min, 20min releasing effect figure.Fig. 3 B are the compounding seas of urase reaction generation
Alginic acid calcisphere embeds small molecule dimethyl diaminophenazine chloride in 0min, 10min, 20min releasing effect figure when having calcium carbonate mixing.Fig. 3 C are
Releasing effect figure of the small molecule dimethyl diaminophenazine chloride in ammonium carbonate/calcium chloride directly reacts the compound calcium alginate microballoon of formation (causes
During close calcium carbonate mixing).As a result one kind can be prepared by the method for the present invention by illustrating changes its release by adjusting calcium carbonate
The complex microsphere of effect.Sodium alginate in this experiment, calcium chloride, sodium carbonate and urase concentration are 5mgmL respectively-1, 20mg
mL-1, 20mgmL-1, 5mgmL-1.Fig. 4 experimental data illustrates calcium carbonate of the present invention/calcium alginate compounded micro-
Ball introduces double-aqueous phase system in preparing, and utilizes aqueous two-phase macromolecular steric effect and specific selectivity so that drop template
Obtained complex microsphere has high envelop rate for the protein substance of macromolecular, and embodiment measures BSA envelop rate
For 97.27%.Sodium alginate in this experiment, calcium chloride concentration are 5mgmL respectively-1, 20mgmL-1。
Fig. 5 data illustrate that by adjusting different enzyme reaction condition-urase concentration different calcium carbonate/alginic acids can be prepared
The microballoon of calcium Compound Degree, thus it is possible to vary the compactness extent of calcium carbonate on calcium carbonate/calcium alginate microsphere, and obtain different
Drug release rate.
A kind of calcium carbonate/calcium alginate that adjustable drug release rate is prepared based on PEG/Dex aqueous two-phase biomineralization technologies
The preparation method of complex microsphere, it is comprised the following steps that：
First step：The preparation of PEG-Dex aqueous two-phases：
(1) mass fraction 10-20% PEG solution, molecular weight 8kDa are configured to；
(2) mass fraction 10-20% Dex solution, molecular weight 500kDa are configured to；
(3) take step (1) and step (2) solution to be sufficiently mixed, stand split-phase, obtain phase and be rich in PEG, lower phase is rich in
Dex aqueous two-phase solution；Extract PEG solution respectively and Dex solution is standby into two beakers；
Second step：Drop template prepares calcium carbonate/calcium alginate compounded microballoon：
(1) solution A：0.01-0.10g sodium alginates and 0.002-0.050g urases are weighed, is added to the above-mentioned of 5-10mL
In Dex solution, it is well mixed, set aside for use；
(2) B solution：Anhydrous calcium chloride 0.1-0.5g and urea 0.1-0.5g are dissolved in 5-10mL PEG solution,
It is well mixed, set aside for use；
(3) solution A is sucked in common commercially available syringe, by tip internal diameter is 20-80 μm with commercial silica gel pipe, its pipe shaft
Internal diameter is that 500 μm of capillary is connected with commercially available syringe, and solution A is advanced in tip capillary by syringe, and will
Solution A is vertically added dropwise in B solution by the way that tip capillary is hanging with the speed of 2 seconds one drops, and its capillary tip is apart from liquid level
0.5-5cm, a diameter of 200~600 μm of calcium alginate microsphere can be formed after the several seconds；Continue to place 1-10 at 20-60 DEG C
After h, treat that enzymatic reaction is fully carried out, produce calcium carbonate/calcium alginate compounded microballoon of adjustable drug release rate.
Embodiment 1
Calcium carbonate/calcium alginate compounded microballoon (enzyme concentration 5mgmL-1, survey 1mgmL-1BSA envelop rate)
1. the preparation of polyethylene glycol (PEG)/glucan (Dex) aqueous two-phase
(1) precision weighs PEG 3.2g and is put into 25mL beaker, adds 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the PEG solution that mass fraction is all 16%；
(2) precision weighs Dex 3.2g and is put into 25mL beaker, adds 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the Dex solution that mass fraction is all 16%；
(3) step (1) and each 10mL mixing of step (2) solution are taken, it is molten that the 20mL aqueous two-phases that mass fraction is 8% are made
Liquid, stand, wait layering.
2. drop template prepares calcium carbonate/calcium alginate compounded microballoon
(1) solution A：0.025g sodium alginates, 0.0050g BSA are weighed, is added in 5mL 8%Dex solution, is mixed
Uniformly, 0.025g urases are added, are well mixed, set aside for use.
(2) B solution：Precision weighs anhydrous calcium chloride 0.2g and urea 0.2g and is dissolved in 10mL 8%PEG solution,
It is stand-by.
(3) solution A suction obtained by step (1) with tip internal diameter is in the syringe of 50 μm of syringe needles, with syringe by its
It is slowly dropped into B solution obtained by 1mL to step (2), rapid balling-up.Continue at 40 DEG C after placing 4h, treat that enzymatic reaction is abundant
Carry out, produce embedding BSA calcium carbonate/calcium alginate compounded microballoon.
(4) B solution 5mL is taken, utilizes the absorbance of BSA in ultraviolet specrophotometer determination sample liquid.And then with original BSA
Absorbance contrasts, and the envelop rate for measuring BSA is 95.13%.
Embodiment 2
Calcium carbonate/calcium alginate compounded microballoon (enzyme concentration 5mgmL-1, survey 2mgmL-1BSA envelop rate)
1. the preparation of polyethylene glycol (PEG)/glucan (Dex) aqueous two-phase
(1) precision weighs PEG 3.2g and is put into 25mL beaker, adds 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the PEG solution that mass fraction is all 16%；
(2) precision weighs Dex 3.2g and is put into 25mL beaker, adds 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the Dex solution that mass fraction is all 16%；
(3) step (1) and each 10mL mixing of step (2) solution are taken, it is molten that the 20mL aqueous two-phases that mass fraction is 8% are made
Liquid, stand, wait layering.
2. drop template prepares calcium carbonate/calcium alginate compounded microballoon
(1) solution A：0.025g sodium alginates, 0.010g BSA are weighed, is added in 5mL 8%Dex solution, is mixed
Uniformly, 0.025g urases are added, are well mixed, set aside for use.
(2) B solution：Precision weighs anhydrous calcium chloride 0.2g and urea 0.2g and is dissolved in 10mL 8%PEG solution,
It is stand-by.
(3) solution A suction obtained by step (1) with tip internal diameter is in the syringe of 50 μm of syringe needles, with syringe by its
It is slowly dropped into B solution obtained by 1mL to step (2), rapid balling-up.Continue at 40 DEG C after placing 4h, treat that enzymatic reaction is abundant
Carry out, produce embedding BSA calcium carbonate/calcium alginate compounded microballoon.
(4) B solution 5mL is taken, utilizes the absorbance of BSA in ultraviolet specrophotometer determination sample liquid.And then with original BSA
Absorbance contrasts, and the envelop rate for measuring BSA is 97.31%.
Embodiment 3
Calcium carbonate/calcium alginate compounded microballoon (enzyme concentration 5mgmL-1, survey 3mgmL-1BSA envelop rate)
1. the preparation of polyethylene glycol (PEG)/glucan (Dex) aqueous two-phase
(1) precision weighs PEG 3.2g and is put into 25mL beaker, adds 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the PEG solution that mass fraction is all 16%；
(2) precision weighs Dex 3.2g and is put into 25mL beaker, adds 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the Dex solution that mass fraction is all 16%；
(3) step (1) and each 10mL mixing of step (2) solution are taken, it is molten that the 20mL aqueous two-phases that mass fraction is 8% are made
Liquid, stand, wait layering.
2. drop template prepares calcium carbonate/calcium alginate compounded microballoon
(1) solution A：0.025g sodium alginates, 0.0150g BSA are weighed, is added in 5mL 8%Dex solution, is mixed
Uniformly, 0.025g urases are added, are well mixed, set aside for use.
(2) B solution：Precision weighs anhydrous calcium chloride 0.2g and urea 0.2g and is dissolved in 10mL 8%PEG solution,
It is stand-by.
(3) solution A suction obtained by step (1) with tip internal diameter is in the syringe of 50 μm of syringe needles, with syringe by its
It is slowly dropped into B solution obtained by 1mL to step (2), rapid balling-up.Continue at 40 DEG C after placing 4h, treat that enzymatic reaction is abundant
Carry out, produce embedding BSA calcium carbonate/calcium alginate compounded microballoon.
(4) B solution 5mL is taken, utilizes the absorbance of BSA in ultraviolet specrophotometer determination sample liquid.And then with original BSA
Absorbance contrasts, and the envelop rate for measuring BSA is 97.27%.
Embodiment 4
Calcium carbonate/calcium alginate compounded microballoon (enzyme concentration 5mgmL-1, survey 4mgmL-1BSA envelop rate)
1. the preparation of polyethylene glycol (PEG)/glucan (Dex) aqueous two-phase
(1) precision weighs PEG 3.2g and is put into 25mL beaker, adds 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the PEG solution that mass fraction is all 16%；
(2) precision weighs Dex 3.2g and is put into 25mL beaker, adds 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the Dex solution that mass fraction is all 16%；
(3) step (1) and each 10mL mixing of step (2) solution are taken, it is molten that the 20mL aqueous two-phases that mass fraction is 8% are made
Liquid, stand, wait layering.
2. sessile drop method template prepares calcium carbonate/calcium alginate compounded microballoon
(1) solution A：0.025g sodium alginates, 0.020g BSA are weighed, is added in 5mL 8%Dex solution, is mixed
Uniformly, 0.025g urases are added, are well mixed, set aside for use.
(2) B solution：Precision weighs anhydrous calcium chloride 0.2g and urea 0.2g and is dissolved in 10mL 8%PEG solution,
It is stand-by.
(3) solution A suction obtained by step (1) is delayed with syringe of the internal diameter for 500 μm of syringe needles with syringe
It is slow to instill in B solution obtained by 1mL to step (2), rapid balling-up.Continue at 40 DEG C after placing 4h, treat that enzymatic reaction is fully entered
OK, embedding BSA calcium carbonate/calcium alginate compounded microballoon is produced.
(4) B solution 5mL is taken, utilizes the absorbance of BSA in ultraviolet specrophotometer determination sample liquid.And then with original BSA
Absorbance contrasts, and the envelop rate for measuring BSA is 96.85%.
Embodiment 5
Calcium carbonate/calcium alginate compounded microballoon (enzyme concentration 5mgmL-1, survey 5mgmL-1BSA envelop rate)
1. the preparation of polyethylene glycol (PEG)/glucan (Dex) aqueous two-phase
(1) precision weighs PEG 3.2g and is put into 25mL beaker, adds 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the PEG solution that mass fraction is all 16%；
(2) precision weighs Dex 3.2g and is put into 25mL beaker, adds 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the Dex solution that mass fraction is all 16%；
(3) step (1) and each 10mL mixing of step (2) solution are taken, it is molten that the 20mL aqueous two-phases that mass fraction is 8% are made
Liquid, stand, wait layering.
2. drop template prepares calcium carbonate/calcium alginate compounded microballoon
(1) solution A：0.025g sodium alginates, 0.0250g BSA are weighed, is added in 5mL 8%Dex solution, is mixed
Uniformly, 0.025g urases are added, are well mixed, set aside for use.
(2) B solution：Precision weighs anhydrous calcium chloride 0.2g and urea 0.2g and is dissolved in 10mL 8%PEG solution,
It is stand-by.
(3) solution A suction obtained by step (1) with tip internal diameter is in the syringe of 50 μm of syringe needles, with syringe by its
It is slowly dropped into B solution obtained by 1mL to step (2), rapid balling-up.Continue at 40 DEG C after placing 4h, treat that enzymatic reaction is abundant
Carry out, produce embedding BSA calcium carbonate/calcium alginate compounded microballoon.
(4) B solution 5mL is taken, utilizes the absorbance of BSA in ultraviolet specrophotometer determination sample liquid.And then with original BSA
Absorbance contrasts, and the envelop rate for measuring BSA is 98.03%.
Embodiment 6
Calcium carbonate/calcium alginate compounded microballoon (enzyme concentration 5mgmL-1, encapsulate dimethyl diaminophenazine chloride)
1. the preparation of polyethylene glycol (PEG)/glucan (Dex) aqueous two-phase
(1) precision weighs PEG 3.2g and is put into 25mL beaker, adds 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the PEG solution that mass fraction is all 16%；
(2) precision weighs Dex 3.2g and is put into 25mL beaker, adds 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the Dex solution that mass fraction is all 16%；
(3) step (1) and each 10mL mixing of step (2) solution are taken, it is molten that the 20mL aqueous two-phases that mass fraction is 8% are made
Liquid, stand, wait layering.
2. sessile drop method prepares template calcium carbonate/calcium alginate compounded microballoon
(1) solution A：0.025g sodium alginates, 0.0050g dimethyl diaminophenazine chlorides are weighed, is added in 5mL 8%Dex solution, is mixed
Close uniformly, add 0.025g urases, be well mixed, set aside for use.
(2) B solution：Precision weighs anhydrous calcium chloride 0.2g and urea 0.2g and is dissolved in 10mL 8%PEG solution,
It is stand-by.
(3) solution A suction obtained by step (1) with tip internal diameter is in the syringe of 50 μm of syringe needles, with syringe by its
It is slowly dropped into B solution obtained by step (2), rapid balling-up.Continue at 40 DEG C after placing 4h, treat that enzymatic reaction is fully carried out,
Produce calcium carbonate/calcium alginate compounded microballoon of embedding small molecule dyes.It is embedded in calcium carbonate/calcium alginate compounded microballoon
Dimethyl diaminophenazine chloride release conditions as shown in fig. 1b.
Embodiment 7
Calcium carbonate/calcium alginate compounded microballoon (enzyme concentration 0mgmL-1, encapsulating BSA surveys release rate)
The preparation of 1.PEG-Dex aqueous two-phases
(1) the accurate PEG for weighing 3.2g is put into 25mL beaker, is added 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the PEG solution that mass fraction is all 16%；
(2) the accurate Dex for weighing 3.2g is put into 25mL beaker, is added 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the Dex solution that mass fraction is all 16%；
(3) step (1) and each 10mL mixing of step (2) solution are taken, it is molten that the 20mL aqueous two-phases that mass fraction is 8% are made
Liquid, stand, wait layering.
2. drop template prepares BSA- calcium carbonate/calcium alginate compounded microballoon
(1) solution A：0.025g sodium alginates, 0.010g BSA are weighed, the mass percent for being added to 5mL is 8%
In Dex solution, it is well mixed, adds 0g urases, is well mixed, set aside for use.
(2) B solution：Precision weighs anhydrous calcium chloride 0.2g and urea 0.2g and is dissolved in 10mL mass percent
It is stand-by in 8% PEG solution.
(3) solution A suction obtained by step (1) with tip internal diameter is in the syringe of 50 μm of syringe needles, with syringe by its
It is slowly dropped into B solution obtained by step (2), rapid balling-up.Continue at 40 DEG C after placing 4h, treat that enzymatic reaction is fully carried out,
Obtain the BSA- calcium carbonate/calcium alginate compounded microballoon prepared under the enzyme concentration.
3. calcium carbonate/calcium alginate compounded microballoon release bovine serum albumin.
Obtained BSA- calcium carbonate/calcium alginate compounded microballoon is all placed in vial with cover, adds deionized water 10mL,
Vial room temperature is placed on rotary mixer, holding rotary speed is 10r/min, takes clear liquid in bottle after 2h, and utilization is ultraviolet
BSA absorbance in spectrophotometric determination sample liquid.The release rate for measuring BSA is 70.59%.
Embodiment 8
Calcium carbonate/calcium alginate compounded microballoon (enzyme concentration 1mgmL-1, encapsulating BSA surveys release rate)
The preparation of 1.PEG-Dex aqueous two-phases
(1) the accurate PEG for weighing 3.2g is put into 25mL beaker, is added 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the PEG solution that mass fraction is all 16%；
(2) the accurate Dex for weighing 3.2g is put into 25mL beaker, is added 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the Dex solution that mass fraction is all 16%；
(3) step (1) and each 10mL mixing of step (2) solution are taken, it is molten that the 20mL aqueous two-phases that mass fraction is 8% are made
Liquid, stand, wait layering.
2. drop template prepares BSA- calcium carbonate/calcium alginate compounded microballoon
(1) solution A：0.025g sodium alginates, 0.010g BSA are weighed, the mass percent for being added to 5mL is 8%
In Dex solution, it is well mixed, adds 0.005g urases, is well mixed, set aside for use.
(2) B solution：Precision weighs anhydrous calcium chloride 0.2g and urea 0.2g and is dissolved in 10mL mass percent
It is stand-by in 8% PEG solution.
(3) solution A suction obtained by step (1) with tip internal diameter is in the syringe of 50 μm of syringe needles, with syringe by its
It is slowly dropped into B solution obtained by step (2), rapid balling-up.Continue at 40 DEG C after placing 4h, treat that enzymatic reaction is fully carried out,
Obtain the BSA- calcium carbonate/calcium alginate compounded microballoon prepared under the enzyme concentration.
3. calcium carbonate/calcium alginate compounded microballoon release bovine serum albumin.
Obtained BSA- calcium carbonate/calcium alginate compounded microballoon is all placed in vial with cover, adds deionized water 10mL,
Vial room temperature is placed on rotary mixer, holding rotary speed is 10r/min, takes clear liquid in bottle after 2h, and utilization is ultraviolet
BSA absorbance in spectrophotometric determination sample liquid.The release rate for measuring BSA is 67.37%.
Embodiment 9
Calcium carbonate/calcium alginate compounded microballoon (enzyme concentration 2mgmL-1, encapsulating BSA surveys release rate)
The preparation of 1.PEG-Dex aqueous two-phases
(1) the accurate PEG for weighing 3.2g is put into 25mL beaker, is added 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the PEG solution that mass fraction is all 16%；
(2) the accurate Dex for weighing 3.2g is put into 25mL beaker, is added 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the Dex solution that mass fraction is all 16%；
(3) step (1) and each 10mL mixing of step (2) solution are taken, it is molten that the 20mL aqueous two-phases that mass fraction is 8% are made
Liquid, stand, wait layering.
2. drop template prepares BSA- calcium carbonate/calcium alginate compounded microballoon
(1) solution A：0.025g sodium alginates, 0.010g BSA are weighed, the mass percent for being added to 5mL is 8%
In Dex solution, it is well mixed, adds 0.010g urases, is well mixed, set aside for use.
(2) B solution：Precision weighs anhydrous calcium chloride 0.2g and urea 0.2g and is dissolved in 10mL mass percent
It is stand-by in 8% PEG solution.
(3) solution A suction obtained by step (1) with tip internal diameter is in the syringe of 50 μm of syringe needles, with syringe by its
It is slowly dropped into B solution obtained by step (2), rapid balling-up.Continue at 40 DEG C after placing 4h, treat that enzymatic reaction is fully carried out,
Obtain the BSA- calcium carbonate/calcium alginate compounded microballoon prepared under the enzyme concentration.
3. calcium carbonate/calcium alginate compounded microballoon release bovine serum albumin.
Obtained BSA- calcium carbonate/calcium alginate compounded microballoon is all placed in vial with cover, adds deionized water 10mL,
Vial room temperature is placed on rotary mixer, holding rotary speed is 10r/min, takes clear liquid in bottle after 2h, and utilization is ultraviolet
BSA absorbance in spectrophotometric determination sample liquid.The release rate for measuring BSA is 58.69%.
Embodiment 10
Calcium carbonate/calcium alginate compounded microballoon (enzyme concentration 3mgmL-1, encapsulating BSA surveys release rate)
The preparation of 1.PEG-Dex aqueous two-phases
(1) the accurate PEG for weighing 3.2g is put into 25mL beaker, is added 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the PEG solution that mass fraction is all 16%；
(2) the accurate Dex for weighing 3.2g is put into 25mL beaker, is added 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the Dex solution that mass fraction is all 16%；
(3) step (1) and each 10mL mixing of step (2) solution are taken, it is molten that the 20mL aqueous two-phases that mass fraction is 8% are made
Liquid, stand, wait layering.
2. sessile drop method prepares template BSA- calcium carbonate/calcium alginate compounded microballoon
(1) solution A：0.025g sodium alginates, 0.010g BSA are weighed, the mass percent for being added to 5mL is 8%
In Dex solution, it is well mixed, adds 0.015g urases, is well mixed, set aside for use.
(2) B solution：Precision weighs anhydrous calcium chloride 0.2g and urea 0.2g and is dissolved in 10mL mass percent
It is stand-by in 8% PEG solution.
(3) solution A suction obtained by step (1) with tip internal diameter is in the syringe of 50 μm of syringe needles, with syringe by its
It is slowly dropped into B solution obtained by step (2), rapid balling-up.Continue at 40 DEG C after placing 4h, treat that enzymatic reaction is fully carried out,
Obtain the BSA- calcium carbonate/calcium alginate compounded microballoon prepared under the enzyme concentration.
3. calcium carbonate/calcium alginate compounded microballoon release bovine serum albumin.
Obtained BSA- calcium carbonate/calcium alginate compounded microballoon is all placed in vial with cover, adds deionized water 10mL,
Vial room temperature is placed on rotary mixer, holding rotary speed is 10r/min, takes clear liquid in bottle after 2h, and utilization is ultraviolet
BSA absorbance in spectrophotometric determination sample liquid.The BSA measured release rate is 52.81%.
Embodiment 11
Calcium carbonate/calcium alginate compounded microballoon (enzyme concentration 4mgmL-1, encapsulating BSA surveys release rate)
The preparation of 1.PEG-Dex aqueous two-phases
(1) the accurate PEG for weighing 3.2g is put into 25mL beaker, is added 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the PEG solution that mass fraction is all 16%；
(2) the accurate Dex for weighing 3.2g is put into 25mL beaker, is added 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the Dex solution that mass fraction is all 16%；
(3) step (1) and each 10mL mixing of step (2) solution are taken, it is molten that the 20mL aqueous two-phases that mass fraction is 8% are made
Liquid, stand, wait layering.
2. drop template prepares BSA- calcium carbonate/calcium alginate compounded microballoon
(1) solution A：0.025g sodium alginates, 0.010g BSA are weighed, the mass percent for being added to 5mL is 8%
In Dex solution, it is well mixed, adds 0.020g urases, is well mixed, set aside for use.
(2) B solution：Precision weighs anhydrous calcium chloride 0.2g and urea 0.2g and is dissolved in 10mL mass percent
It is stand-by in 8% PEG solution.
(3) solution A suction obtained by step (1) with tip internal diameter is in the syringe of 50 μm of syringe needles, with syringe by its
It is slowly dropped into B solution obtained by step (2), rapid balling-up.Continue at 40 DEG C after placing 4h, treat that enzymatic reaction is fully carried out,
Obtain the BSA- calcium carbonate/calcium alginate compounded microballoon prepared under the enzyme concentration.
3. calcium carbonate/calcium alginate compounded microballoon release bovine serum albumin.
Obtained BSA- calcium carbonate/calcium alginate compounded microballoon is all placed in vial with cover, adds deionized water 10mL,
Vial room temperature is placed on rotary mixer, holding rotary speed is 10r/min, takes clear liquid in bottle after 2h, and utilization is ultraviolet
BSA absorbance in spectrophotometric determination sample liquid.The release rate for measuring BSA is 48.6%.
Embodiment 12
Calcium carbonate/calcium alginate compounded microballoon (enzyme concentration 5mgmL-1, bag BSA surveys release rate)
The preparation of 1.PEG-Dex aqueous two-phases
(1) the accurate PEG for weighing 3.2g is put into 25mL beaker, is added 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the PEG solution that mass fraction is all 16%；
(2) the accurate Dex for weighing 3.2g is put into 25mL beaker, is added 16.8mL deionized water, is stirred using magnetic force
It is 300r/min to mix device speed, stirs 2h, is configured to the Dex solution that mass fraction is all 16%；
(3) step (1) and each 10mL mixing of step (2) solution are taken, it is molten that the 20mL aqueous two-phases that mass fraction is 8% are made
Liquid, stand, wait layering.
2. drop template prepares BSA- calcium carbonate/calcium alginate compounded microballoon
(1) solution A：0.025g sodium alginates, 0.010g BSA are weighed, the mass percent for being added to 5mL is 8%
In Dex solution, it is well mixed, adds 0.025g urases, is well mixed, set aside for use.
(2) B solution：Precision weighs anhydrous calcium chloride 0.2g and urea 0.2g and is dissolved in 10mL mass percent
It is stand-by in 8% PEG solution.
(3) solution A suction obtained by step (1) with tip internal diameter is in the syringe of 50 μm of syringe needles, with syringe by its
It is slowly dropped into B solution obtained by step (2), rapid balling-up.Continue at 40 DEG C after placing 4h, treat that enzymatic reaction is fully carried out,
Obtain the BSA- calcium carbonate/calcium alginate compounded microballoon prepared under the enzyme concentration.
3. calcium carbonate/calcium alginate compounded microballoon release bovine serum albumin.
Obtained BSA- calcium carbonate/calcium alginate compounded microballoon is all placed in vial with cover, adds deionized water 10mL,
Vial room temperature is placed on rotary mixer, holding rotary speed is 10r/min, takes clear liquid in bottle after 2h, and utilization is ultraviolet
BSA absorbance in spectrophotometric determination sample liquid.The release rate for measuring BSA is 43.25%.
Claims (6)
1. it is a kind of adjustable drug release rate is prepared based on PEG/Dex aqueous two-phase biomineralization technologies calcium carbonate/calcium alginate it is multiple
Close microballoon, it is characterised in that calcium carbonate/calcium alginate compounded microballoon is shell solid construction, and its shell is calcium alginate and carbonic acid
The compound to be formed is blended in calcium, and its layer is that the calcium carbonate that mineralising reaction is formed is grown on grid in the layer of calcium alginate microsphere, and
The protein drug of macromolecular can be encapsulated；Complex microsphere particle diameter distribution is 200~600 μm.
2. the calcium carbonate of adjustable drug release rate according to claim 1/calcium alginate compounded microballoon, it is characterised in that
Selective partition characteristic using the aqueous two-phase to macro-molecular protein, the protein drug of macromolecular is encapsulated in described
The interior phase of Dex aqueous two-phase drop templates.
3. the calcium carbonate of adjustable drug release rate according to claim 1 or 2/calcium alginate compounded microballoon, its feature exist
In, using the Dex drops as template using drop template formed microballoon；And then macro-molecular protein class medicine is being solidified
It is encapsulated in during regulation in compound calcium alginate microballoon.
4. the calcium carbonate of adjustable drug release rate according to claim 3/calcium alginate compounded microballoon, it is characterised in that
The microballoon of different calcium carbonate/calcium alginate compounded degree can be prepared by the enzyme reaction condition-urase concentration for adjusting different, and obtained
Obtain different drug release rates.
5. the calcium carbonate of the adjustable drug release rate according to claim 1 or 2 or 3/calcium alginate compounded microballoon, its feature
It is, the aqueous two-phase drop template, is that a certain amount of urea and a certain amount of calcium chloride are scattered in 8% polyethylene glycol phase PEG
In (molecular weight 8kDa), a certain amount of urase and a certain amount of sodium alginate are scattered in 8% glucan phase Dex (molecular weight 500kDa)
In；Dex phases are instilled by the tip capillary that tip internal diameter is 20-80 μm calcium carbonate/calcium alginate compounded is formed in PEG phases
Microballoon.
6. it is a kind of adjustable drug release rate is prepared based on PEG/Dex aqueous two-phase biomineralization technologies calcium carbonate/calcium alginate it is multiple
Close the preparation method of microballoon, it is characterised in that concretely comprise the following steps：
First step：The preparation of PEG-Dex aqueous two-phases：
(1) mass fraction 10-20% PEG solution, molecular weight 8kDa are configured to；
(2) mass fraction 10-20% Dex solution, molecular weight 500kDa are configured to；
(3) take step (1) and step (2) solution to be sufficiently mixed, stand split-phase, obtain phase and be rich in PEG, lower phase is rich in Dex's
Aqueous two-phase solution；Extract PEG solution respectively and Dex solution is standby into two beakers；Second step：Drop template prepares carbon
Sour calcium/calcium alginate compounded microballoon：
(1) solution A：0.01-0.10g sodium alginates and 0.002-0.050g urases are weighed, the above-mentioned Dex for being added to 5-10mL is molten
In liquid, it is well mixed, set aside for use；
(2) B solution：Anhydrous calcium chloride 0.1-0.5g and urea 0.1-0.5g are dissolved in 5-10mL PEG solution, mixing is equal
It is even, set aside for use；
(3) solution A is sucked in common commercially available syringe, by tip internal diameter is 20-80 μm with commercial silica gel pipe, its tube body diameter
It is connected with commercially available syringe for 500 μm of capillaries, solution A is advanced in tip capillary by syringe, and by solution A
Vertically it is added dropwise to by the way that tip capillary is hanging with the speed of 2 seconds one drops in B solution, its capillary tip is apart from liquid level 0.5-
5cm, a diameter of 200~600 μm of calcium alginate microsphere can be formed after the several seconds；Continue at 20-60 DEG C after placing 1-10h,
Treat that enzymatic reaction is fully carried out, produce calcium carbonate/calcium alginate compounded microballoon of adjustable drug release rate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710768009.0A CN107789332B (en) | 2017-08-31 | 2017-08-31 | Calcium carbonate/calcium alginate composite microspheres capable of adjusting drug release rate and prepared based on double-aqueous-phase biomineralization technology |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710768009.0A CN107789332B (en) | 2017-08-31 | 2017-08-31 | Calcium carbonate/calcium alginate composite microspheres capable of adjusting drug release rate and prepared based on double-aqueous-phase biomineralization technology |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107789332A true CN107789332A (en) | 2018-03-13 |
| CN107789332B CN107789332B (en) | 2020-01-14 |
Family
ID=61532246
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710768009.0A Active CN107789332B (en) | 2017-08-31 | 2017-08-31 | Calcium carbonate/calcium alginate composite microspheres capable of adjusting drug release rate and prepared based on double-aqueous-phase biomineralization technology |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107789332B (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108514896A (en) * | 2018-03-23 | 2018-09-11 | 西南交通大学 | A kind of preparation method and device of micro-fluidic aqueous two-phase monodisperse calcium alginate microsphere |
| CN109701461A (en) * | 2018-10-25 | 2019-05-03 | 西南交通大学 | A kind of preparation and its application of the calcium carbonate based on PEG/Dex aqueous two-phase/calcium alginate compounded micro-capsule |
| CN109943558A (en) * | 2019-04-22 | 2019-06-28 | 西南交通大学 | A method of based on double-aqueous phase system biomimetic mineralization process immobilised enzymes |
| CN110343288A (en) * | 2019-07-19 | 2019-10-18 | 西南交通大学 | It is a kind of using aqueous two-phase lotion as the porous calcium alginate microsphere of template, preparation method and applications |
| CN111077182A (en) * | 2019-12-31 | 2020-04-28 | 东南大学 | Method for reducing nanobelt contact thermal resistance by liquid assistance |
| CN111304189A (en) * | 2020-02-28 | 2020-06-19 | 西南交通大学 | Enzyme-loaded calcium alginate microsphere enhanced cascade enzymatic reaction method based on aqueous two-phase system |
| CN113634208A (en) * | 2021-08-20 | 2021-11-12 | 西南交通大学 | Method for preparing porous calcium alginate microspheres by using microfluidic double-aqueous-phase emulsion as template |
| WO2022000304A1 (en) * | 2020-06-29 | 2022-01-06 | 江苏苏博特新材料股份有限公司 | Microcapsule type polycarboxylate superplasticizer and preparation method therefor |
| CN118561557A (en) * | 2024-07-30 | 2024-08-30 | 浙江大学 | Active and passive integrated repair mortar for leakage tunnel lining and construction process |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06240040A (en) * | 1993-02-16 | 1994-08-30 | Dainichiseika Color & Chem Mfg Co Ltd | Composite microballoon and its production |
| CN101081911A (en) * | 2007-06-15 | 2007-12-05 | 天津大学 | Big molecular engram calcium orthophosphate/calcium alginate hybridized micro-balloon and method for preparing the same |
| CN101092494A (en) * | 2007-06-15 | 2007-12-26 | 天津大学 | Hybridized microballons of calcium polyacrylate / calcium alginate of macro molecule engram |
| CN102698667A (en) * | 2012-06-19 | 2012-10-03 | 重庆大学 | Spherical pore foaming agent with nuclear shell structure and three-dimensional cytoskeleton prepared by same |
-
2017
- 2017-08-31 CN CN201710768009.0A patent/CN107789332B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH06240040A (en) * | 1993-02-16 | 1994-08-30 | Dainichiseika Color & Chem Mfg Co Ltd | Composite microballoon and its production |
| CN101081911A (en) * | 2007-06-15 | 2007-12-05 | 天津大学 | Big molecular engram calcium orthophosphate/calcium alginate hybridized micro-balloon and method for preparing the same |
| CN101092494A (en) * | 2007-06-15 | 2007-12-26 | 天津大学 | Hybridized microballons of calcium polyacrylate / calcium alginate of macro molecule engram |
| CN102698667A (en) * | 2012-06-19 | 2012-10-03 | 重庆大学 | Spherical pore foaming agent with nuclear shell structure and three-dimensional cytoskeleton prepared by same |
Non-Patent Citations (1)
| Title |
|---|
| 赵武奇等: "壳聚糖/海藻酸钠微球对红景天苷控制释放的研究", 《吉林农业大学学报》 * |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108514896A (en) * | 2018-03-23 | 2018-09-11 | 西南交通大学 | A kind of preparation method and device of micro-fluidic aqueous two-phase monodisperse calcium alginate microsphere |
| CN109701461A (en) * | 2018-10-25 | 2019-05-03 | 西南交通大学 | A kind of preparation and its application of the calcium carbonate based on PEG/Dex aqueous two-phase/calcium alginate compounded micro-capsule |
| CN109701461B (en) * | 2018-10-25 | 2021-06-08 | 西南交通大学 | Preparation and application of PEG/Dex double aqueous phase-based calcium carbonate/calcium alginate composite microcapsule |
| CN109943558A (en) * | 2019-04-22 | 2019-06-28 | 西南交通大学 | A method of based on double-aqueous phase system biomimetic mineralization process immobilised enzymes |
| CN109943558B (en) * | 2019-04-22 | 2023-02-17 | 西南交通大学 | Method for immobilizing enzyme based on biomimetic mineralization process of double-aqueous-phase system |
| CN110343288B (en) * | 2019-07-19 | 2021-06-08 | 西南交通大学 | Porous calcium alginate microspheres using aqueous two-phase emulsion as template, preparation method and application thereof |
| CN110343288A (en) * | 2019-07-19 | 2019-10-18 | 西南交通大学 | It is a kind of using aqueous two-phase lotion as the porous calcium alginate microsphere of template, preparation method and applications |
| CN111077182A (en) * | 2019-12-31 | 2020-04-28 | 东南大学 | Method for reducing nanobelt contact thermal resistance by liquid assistance |
| CN111077182B (en) * | 2019-12-31 | 2022-04-08 | 东南大学 | Method for reducing nanobelt contact thermal resistance by liquid assistance |
| CN111304189A (en) * | 2020-02-28 | 2020-06-19 | 西南交通大学 | Enzyme-loaded calcium alginate microsphere enhanced cascade enzymatic reaction method based on aqueous two-phase system |
| WO2022000304A1 (en) * | 2020-06-29 | 2022-01-06 | 江苏苏博特新材料股份有限公司 | Microcapsule type polycarboxylate superplasticizer and preparation method therefor |
| CN113929342A (en) * | 2020-06-29 | 2022-01-14 | 南京博特新材料有限公司 | Microcapsule type polycarboxylic acid superplasticizer and preparation method thereof |
| CN113634208A (en) * | 2021-08-20 | 2021-11-12 | 西南交通大学 | Method for preparing porous calcium alginate microspheres by using microfluidic double-aqueous-phase emulsion as template |
| CN118561557A (en) * | 2024-07-30 | 2024-08-30 | 浙江大学 | Active and passive integrated repair mortar for leakage tunnel lining and construction process |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107789332B (en) | 2020-01-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107789332A (en) | A kind of calcium carbonate/calcium alginate compounded microballoon that adjustable drug release rate is prepared based on aqueous two-phase biomineralization technology | |
| US4933185A (en) | System for controlled release of biologically active compounds | |
| US5427935A (en) | Hybrid membrane bead and process for encapsulating materials in semi-permeable hybrid membranes | |
| CN109701461B (en) | Preparation and application of PEG/Dex double aqueous phase-based calcium carbonate/calcium alginate composite microcapsule | |
| CA2874374C (en) | Micro-particles, blood-substitute and method for forming same | |
| CN104762289B (en) | The method that microporous membrane permeation emulsification prepares the gelatine microsphere of fixed alcohol dehydrogenase | |
| CA1184518A (en) | Reversible microencapsulation | |
| CA1230054A (en) | Sustained release semi-permeable capsule | |
| CA1245984A (en) | Polyionic microencapsulation | |
| CA1172586A (en) | Method of culturing anchorage dependent cells | |
| CN106582465A (en) | Method for preparing chitosan/sodium alginate natural polymer core-shell microspheres by one-step process | |
| CN109897387A (en) | Application, porous gel and its preparation of a kind of modified gelatin in water packet air lotion | |
| CN109054846B (en) | Preparation method of soil conditioner based on layer-by-layer self-assembled microspheres | |
| CN106076214A (en) | A kind of calcium alginate microsphere preparation method with nucleocapsid structure | |
| CN107198791A (en) | The method that electrostatic spraying prepares cross linked porous starch hemostatic microsphere | |
| CN105296460A (en) | Microbial capsule for wastewater treatment and preparation method thereof | |
| CN104624129A (en) | Preparation method of starch nanometer microspheres based on ionic liquid-type surfactant microemulsion system | |
| CN107970228A (en) | A kind of preparation method using chitosan-TPP-KGM as the nano-microcapsule of compound wall materials | |
| WO1989001034A1 (en) | Encapsulation of biological materials in semi-permeable membranes | |
| CN101401958B (en) | Chitosan-hydroxyapatite composite spheroidal particle, producing method and apparatus thereof | |
| JPS59205985A (en) | Recovery of non-secretory substance produced from cell | |
| Meng et al. | Poly (vinyl alcohol)/sodium alginate polymer membranes as eco‐friendly and biodegradable coatings for slow release fertilizers | |
| CN106345419B (en) | A kind of preparation method of the calcium alginate microsphere with porous structure | |
| CN100334036C (en) | Composite microsphere containing hydroxyapatite and preparing method thereof | |
| CN105838013B (en) | One kind is based on methyl vinyl ether maleic acid copolymer and the sensitive composite Nano gels of chitosan pH and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |