CN108047484A - The preparation method and application of camptothecine microballoon - Google Patents
The preparation method and application of camptothecine microballoon Download PDFInfo
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Abstract
The invention discloses a kind of preparation method and applications of camptothecine microballoon.For the present invention using methanol, chloroform mixed solvent as pore-foaming agent, MAA is function monomer, and camptothecine is template molecule, and EGDMA is crosslinking agent, and azodiisobutyronitrile is initiator, and camptothecine microballoon is made in water-bath vibration polymerization.Synthesizing obtained microballoon has higher adsorptivity, and in the case where concentration is 1mmol/L, adsorbance is up to 62.45 μm of ol/g；With good affine performance, higher Selective adsorption is shown, separation factor α is up to 2.09.Camptothecin molecule trace micro-sphere method prepared by the present invention is simple for process, and good high selection sorptive material can be provided for camptothecin analogues.
Description
Technical field
The present invention relates to the preparing technical field of molecular engram microsphere, more particularly, to a kind of system of camptothecine microballoon
Preparation Method and application.
Background technology
Camplotheca acuminata is south China specialty plant, and natural resources are extremely limited, and camptothecine has unique anticancer mechanism, clinical
The upper treatment for kinds cancer, content of the camptothecine in camplotheca acuminata is very low, and the content highest wherein in fruit of camptotheca acuminata is
0.02%, general yield about 0.008% in root.Low content in addition natural products ingredient complexity to camptothecine extraction and point
From bringing big inconvenience.Camptothecine content in camplotheca acuminata is relatively low, system is complicated, separation and concentration difficulty is big while water solubility is low,
The features such as effective pharmacophoric group stability is poor, bioavilability is poor limits its clinical research and development.
The development prospect that molecular imprinting technology will have in Separation of Natural Products and purifying.Theoretically if analysis can answer
With the molecular imprinting technology with particular molecule identification function, it is polymerize by synthesizing the molecular engram with camptothecine template molecule
Object obtains the hole exactly matched with camptothecine and the like function base after eluted template molecule, then is expected to using efficiently point
Technology from purifying solves the problems, such as the separating-purifying of camptothecine, to obtain the high anti-cancer activity of high-purity, wide anticancer spectrum
Camptothecin analogues.But have no at present can specific aim and high efficiency be applied to camptothecin analogues separating-purifying application
Correlation molecule trace object correlative study report.
The content of the invention
It is micro- the technical problem to be solved by the present invention is to be directed to existing camptothecin analogues separating-purifying molecular engram
The technical deficiency of ball provides a kind of preparation method of camptothecine microballoon.
Another technical problems to be solved of the present invention are to provide the camptothecine microballoon that the method is prepared.
A present invention also technical problems to be solved are to provide the camptothecine microballoon as camptothecine separating-purifying use point
Application in terms of sub- imprinted polymer.
The purpose of the present invention is achieved by the following technical programs：
A kind of preparation method of camptothecine microballoon is provided, is using the mixed solvent of methanol and chloroform as pore-foaming agent, MAA is work(
Energy monomer, camptothecine is template molecule, and EGDMA is crosslinking agent, under the action of initiator, dry after water-bath vibration polymerization,
It is obtained after elution.
The present invention can be achieved by the way that the amount of methanol in solvent is controlled to control the grain size of microballoon.Methanol content it is more more more
The easily microballoon of generation bulky grain.Preferably, the volume ratio of the in the mixed solvent, methanol and chloroform is 5~8:5~2.Into one
The volume ratio of the preferred the two of step is 6:4.
The dosage and reaction temperatures affect of the mixed solvent the mobile performance of polymerization system, too small dosage or excessively high
Temperature will cause system mobility reduce, product caking, and then influence microballoon absorption property.Preferred solvent dosage be by
According to the gauge of 1moL template molecules, the solvent of addition is 15~40mL, is most preferably that solvent dosage is 30mL.
Preferably, the temperature of the polymerization is 50 DEG C~60 DEG C, further preferred 60 DEG C.
Preferably, when the time of the polymerization is 18 small.
Preferably, the dosage of the function monomer is 2~8 according to the mol ratio of itself and template molecule:1 determines.Into one
The mol ratio for walking preferred function monomer and template molecule is 4:1 or 5:1.
Preferably, the dosage of the crosslinking agent is 10~30 according to the mol ratio of itself and template molecule:1 determines.Into one
The mol ratio for walking preferred crosslinking agent and template molecule is 15:1.
It is highly preferred that the mol ratio of the template molecule, function monomer, crosslinking agent is 1:5:15.Described preferred
Under the conditions of optimum proportioning, the microballoon polymerizeing has higher adsorptivity, and in the case where concentration is 1mmol/L, adsorbance is reachable
62.45μmol/g.Preferably, initiator uses azodiisobutyronitrile (AIBN) or azo diamyl cyanogen (ABDV).Further preferably
Ground, under the conditions of template molecule is 1mol, the dosage of the initiator is 0.1g.
Preferably, the elution is using V (methanol):V (acetic acid)=9:1 mixed solution is washed off using soxhlet extraction method
Template molecule and unreacted substance.
Currently preferred preparation method comprises the following steps：
S1. weigh a certain amount of template molecule camptothecine and function monomer MAA is dissolved in a certain amount of solvent, room temperature is shaken
Prepolymerization is shaken, camptothecine is completely dissolved and forms stable hydrogen bond with MAA；
S2. crosslinking agent EGDMA and initiator A IBN is added in into the system after step S1 prepolymerizations, leads to N2Deoxidation, sealing
After water bath with thermostatic control vibration polymerization under certain temperature；
S3. polymerizate taking-up is cooled to room temperature after the completion of the polymerisation described in step S2, centrifugal drying must polymerize
Object；
S4. eluted template molecule and unreacted substance, until it can't detect template molecule in eluent, then with molten
Agent washes away excessive acetic acid, is dried in vacuo to get to camptothecine microballoon namely camptothecine imprinted polymer (MIPs).
Preferably, solvent described in step S1 is mixed solvent, be methanol and chloroform according to volume ratio is 5~8:5~2 ratio
Example determines.
Preferably, when the prepolymerized time described in step S1 is 1 small.
Preferably, the temperature of water bath with thermostatic control is 50 DEG C~60 DEG C, further preferred 60 DEG C under certain temperature described in step S2.
Preferably, when the time polymerizeing described in step S2 is 18 small.
Preferably, the logical N2The time of deoxidation is 10min.
Preferably, elution described in step S4 is using V (methanol):V (acetic acid)=9:1 mixed solution uses soxhlet extraction
Method washes off template molecule and unreacted substance.Solvent described in step S4 is methanol.
What present invention offer the method was prepared obtains camptothecine microballoon.
Present invention simultaneously provides application of the camptothecine microballoon in terms of camptothecine is adsorbed, and printed particular as camptothecine
Application of the mark polymer (MIPs) in terms of separating-purifying camptothecine.
Beneficial effects of the present invention are as follows：
The present invention provides a kind of methods for preparing camptothecine microballoon, are using the mixed solvent of methanol and chloroform as pore
Agent, MAA are function monomer, and camptothecine is template molecule, and EGDMA is crosslinking agent, under the action of initiator, are vibrated through water-bath poly-
It being obtained after dry after conjunction, elution, the camptothecine microballoon being prepared can be applied as microsphere material in chromatographic stationary phases,
Efficient molecule distinguishability is shown, effective camplotheca acuminata alkali composition in complex extraction object is adsorbed, so as to reach what is isolated and purified
Purpose.
The present invention uses MAA to obtain excellent polymerization effect for function monomer.The molecule knot of methacrylic acid (MAA) class
Contain in structure there are one carbon-carbon double bond and a carboxyl, hydrogen bond can be generated with amino or carboxyl compound, it also can be in certain condition
Lower and amine substance generates ionization, and higher choosing is shown as the molecularly imprinted polymer that function monomer synthesizes using it
Select adsorptivity.
The present invention is realized creatively using the mixed solvent of methanol and chloroform as polymerisation by controlling mixing
The amount of the methanol added in solvent controls the grain size of microballoon, for the extensive use of camptothecine microballoon provides strong technology branch
It holding, the present invention further summarizes preferred solvent dosage and polymerization temperature, effectively controls the mobile performance of system in polymerisation,
So as to regulate and control the optimal absorption property of microballoon.
The present invention determines adsorption isotherms of the MIPs under different camplotheca acuminata alkali concns, and MIPs has good compatibility
Can, while probed into its absorption property with Scathard analytic approach.The molecularly imprinted polymer that present invention synthesis obtains is to mesh
Molecule tool is marked there are two types of binding site, by using hydroxycamptothecin the making choice property adsorption experiment similar with camptothecine structure
In, molecular engram microsphere shows Selective adsorption more higher than blank trace microballoon, and separation factor α is up to 2.09.With this hair
Camptothecin molecule trace microballoon prepared by bright method is simple for process, and good high selection can be provided for camptothecin analogues and is inhaled
Attached property material.
Description of the drawings
Standard curve of Fig. 1 camptothecines under 1~10 μ g/mL concentration.
The Dynamic Adsorption curve of Fig. 2 camptothecines MIPs.
The microballoon SEM figures of Fig. 3 different solvents ratio synthesis.
Influence of Fig. 4 different solvents dosage to the adsorption capacity of microballoon.
Influence of Fig. 5 function monomers dosage to the absorption property of microballoon.
The microballoon SEM figures of Fig. 6 difference in functionality monomer ratio synthesis.
Influence of the dosage of Fig. 7 crosslinking agents to microballoon absorption property.
The adsorption isotherm of Fig. 8 MIPs and NIPs.
Fig. 9 MIPs and NIPs is to the Scatchard analysis charts of the absorption property of camplotheca acuminata aqueous slkali.
Specific embodiment
It is further illustrated the present invention with reference to specific embodiment.Following embodiments are only for illustration, it is impossible to manage
It solves as limitation of the present invention.Unless stated otherwise, the reagent used in following embodiments is that conventional purchased in market or commercial sources obtain
The reagent obtained, unless stated otherwise, the method and apparatus used in following embodiments is method commonly used in the art and sets
It is standby.
Embodiment 1
1. experiment reagent
Camptothecine (analyzes pure, Jinan Dai Zheng Co., Ltds)；Camptothecine, hydroxycamptothecin (standard items, Chinese measuring science
Research institute)；α-methacrylic acid (MAA) (Tianjin Kermel Chemical Reagent Co., Ltd.)；2 ' 2- azodiisobutyronitriles
(AIBN, 98%), ethylene glycol dimethacrylate (EGDMA, 98%) (Aladdin reagent)；Acetonitrile, methanol, chloroform, ice second
Acid (analyzes pure, Tianjin great Mao chemical reagent factories).
2. laboratory apparatus
THZ-82 water-bath constant temperature oscillators, high honour instrument manufacturing Co., Ltd；KQ-300B type ultrasonic cleaners, Kunshan
Ultrasonic instrument Co., Ltd of city；UV-2450 ultraviolet specrophotometers, agilent company；Environmental scanning electron microscope
(SEM), the South China Botanical Garden Chinese Academy of Sciences；High performance liquid chromatograph, agilent company.
3. the synthesis microballoon of camptothecin molecule trace microballoon
It weighs a certain amount of template molecule camptothecine and function monomer MAA is dissolved in a certain amount of solvent, rocked at room temperature
Prepolymerization 1h is completely dissolved camptothecine and forms stable hydrogen bond with MAA, adds in crosslinking agent EGDMA and initiator A IBN, lead to
N2Deoxidation 10min is sealed after water bath with thermostatic control vibration polymerization 18h under certain temperature.Polymerizate taken out after the completion of reaction cold
But to room temperature, centrifugal drying obtains polymer.With V (methanol):V (acetic acid)=9:1 mixed solution washes off mould using soxhlet extraction method
Plate molecule and unreacted substance, until it can't detect template molecule in eluent, again with methanol washes away excessive acetic acid,
Vacuum drying is to get to camptothecine imprinted polymer (MIPs).
Under conditions of template molecule is not added with, blank imprinted polymer (NIPs) is made in the same way.
The influence under different synthesis conditions to microballoon equilibrium adsorption capacity (Q), microballoon pattern is investigated in experiment using single_factor method,
Main limit of consideration：
1. solvent selects：Acetonitrile, methanol, chloroform, methanol/chloroform mixed solvent
2. solvent dosage：15mL、20mL、30mL、40mL
3. mixed solvent ratio：V (methanol：Chloroform)：5:5、6:4、7:3、8:2
4. synthesis temperature：50℃、55℃、60℃
5. function monomer dosage：Mol (camptothecines：MAA)：1:2、1:4、1:6、1:8
6. dosage of crosslinking agent：Mol (camptothecines：EGDMA)：1:10、1:15、1:20、1:30
1 different condition of table synthesizes camptothecin molecule trace microballoon
4.HPLC measures camptothecine content
(1) HPLC chromatogram condition：Experiment measures camptothecine content using high performance liquid chromatography, and chromatographic condition is：Chromatographic column：
Kromasil 100-5C18 (4.6mm × 250mm, 5um)；Column temperature：25℃；Flow velocity：1mL/min；Mobile phase：Acetonitrile/water (40/
60, V:V)；Sample size：20μL；Detection wavelength：254nm.It is filtered before sample feeding through 0.45 μm of organic phase filter membrane.
(2) preparation of standard solution：Camptothecin standard product 0.0034g accurately is weighed, uses methanol：Acetonitrile (1:1, V/V) it is molten
Liquid dissolves and is settled to 10mL, obtains the camplotheca acuminata aqueous slkali of 0.976mmol/l.
(3) the Dynamic Adsorption experiment of imprinted polymer：
Six parts of 5mg camptothecin molecules imprinted polymers (MIPs) are weighed respectively, add in the camptothecine first of 10mL1mmol/L
Alcohol:Acetonitrile (1:1, V/V) in solution, take out, centrifuge, organic phase is through 0.45 μm after shaken at room temperature 2h, 4h, 6h, 8h, 10h, 12h
After membrane filtration, high performance liquid chromatography measures C before absorption1, C after absorption2The concentration of camptothecine, calculates Dynamic Adsorption in solution
Test the adsorption capacity Q of each period.
In formula (I)：
C1：Concentration (mmol/g) before absorption
C2:Concentration (mmol/g) after absorption
V:Adsorbent solution volume (mL)
m：Polymer quality (mg)
(4) equilibrium adsorption experiment
5mg polymer is weighed, adds in the camptothecine methanol of 10mL, 1mmol/L：Acetonitrile (1:1, V/V) chromatographically pure solution, room
Temperature vibration centrifuges solution after must adsorbing after a certain period of time, and the content of camptothecine in solution is measured by high performance liquid chromatography.Pass through
Formula (I) calculates equilibrium adsorption capacities Q, and camptothecin molecule trace microballoon is synthesized come each factor of comparison by adsorption capacity values
It influences.
(5) adsorption isotherm
It is accurate to weigh each 5mg of MIPs and NIPs, add in the camptothecine methanol of certain volume various concentration：Chloroform (1:1, V/
V) chromatographically pure solution, at room temperature vibration absorption certain time, centrifugation, takes supernatant liquor to be measured by high performance chromatograph in solution and likes
Set the content of alkali.Adsorption capacity Q is calculated, and adsorption isotherm is drawn according to the variation of adsorption capacity.
(6) the Selective adsorption experiment of microballoon
The competitive Adsorption experiment of MIPs：Camptothecine, the hydroxycamptothecin chromatographically pure solution of isoconcentration are prepared respectively, it is each to add in
The MIPs and NIPs of 5mg, to measure its dense for high performance liquid chromatography under 254nm, 266nm respectively after a certain period of time for shaken at room temperature absorption
Degree compares the adsorbance of the two.
(7) scanning electron microscope
The microballoon of the drying prepared is sticked on double faced adhesive tape, vacuum metal spraying, observed with scanning electron microscope (SEM) micro-
Spherical looks, and from SEM figures measure 50~100 microballoons grain size, calculate average grain diameter.
5. experimental result
(1) drafting of camptothecin standard curve
Measure camptothecine using high performance liquid chromatography is in the linear equation of methanol/acetonitrile mixed solution：Y=91.776x+
4.342,R2=0.9998.Standard curve of the camptothecine under 1~10 μ g/mL concentration is as shown in Figure 1.
(2) the Dynamic Adsorption curve of imprinted polymer
Dynamic Adsorption curve is made as shown in Figure 2 in adsorption capacity in different time periods according to MIPs.Experiment is found
MIPs initial absorbing rates are very fast, and maximal absorptive capacity is reached after 4h, and as the time increases, adsorbance, which tapers into, finally to exist
10~12h tends towards stability.The reason is that since saturation has not yet been reached in the binding site in microballoon when just starting absorption, still have and mould
The ability that plate molecule combines, with the increase of adsorption time, adsorbance also gradually increases；When adsorption time reaches 4h, polymerization
Object surface void has been occupied, and the camptothecin molecule to dissociate in solution, which reaches internal cavity, needs the longer time, while when molten
The concentration of camptothecine reduces in liquid, and outer the phenomenon that spitting can occur for the polymer that part surface has adsorbed template molecule so that absorption
Amount reduces.Dynamic Adsorption finally reaches adsorption equilibrium after 10~12h.
(3) optimizing research of camptothecin molecule trace microballoon synthesis condition and experiment
Solvent determines：
Experiment finds that solubility of the camptothecine in all kinds of solvents is relatively low, can only be dissolved in pyridine, chloroform, dichloromethane, first
Alcohol/chloroform mixed solution etc..Lot of experiments is summarized, it is found that camptothecine dissolubility in chloroform, dichloromethane is best.Initial stage
Polymerisation is carried out using single chloroformic solution, but because chloroform is volatile, volatilization is easy in heat polymerization process completely, is led
Cause the product ultimately generated caking, it is difficult to collect.Therefore a certain proportion of methanol is creatively added in system, increase solution pole
Property, reduce solution evaporation amount in reaction.
In the case where other conditions are constant, polymerization temperature is 60 DEG C, and camplotheca acuminata alkali concn is 4.167mmol/L, and template is divided
Son：Function monomer：Crosslinker ratio is 1:4:20, change the ratio of methanol and chloroform, by compare equilibrium adsorption capacities Q and
Pass through the change of size of scanning electron microscopic observation microballoon.
Fig. 3 show the microballoon SEM figures of different solvents ratio synthesis.It can be seen that suction of the change of solvent ratios to microballoon
Attached performance influence is simultaneously little, but has been largely fixed the size of microspherulite diameter.When methanol content is more, polymerization system
Polarity, viscosity and surface tension it is bigger, the mass transfer rate of system is caused to reduce, the aggregation velocity of polymer segment slows down, gather
The surface tension increase that synthesis ball needs overcome, the MIPs grain sizes of formation are smaller.Conversely, methanol content is reduced, chloroform content increases
Add, polarity, the surface tension of polymerization system reduce, and are more advantageous to the formation of big grain size MIPMs.Chloroform and methanol in polymerization system
Addition be the key that influence MIPs particle sizes, methanol is unfavorable for the balling-up of big ball, and chloroform is then conducive to generate big grain
The MIPs in footpath.Grain size is smaller, and microsphere surface product is bigger, can definitely increase with respect to its absorption property.It is closed using precipitation polymerization method
The ratio of methanol and chloroform according to practical application, can be changed into camptothecine microballoon, to achieve the purpose that control the size of microballoon.
Influence of the 2 different solvents ratio of table to microballoon absorption property
Solvent dosage determines：
Keep other conditions constant, the present embodiment is with the preferable value range of solvent dosage that numerous studies of the present invention determine
In 15mL~40mL, change solvent dosage 15mL, 20mL, 30mL, 40mL as representative, the concentration for being corresponding in turn to camptothecine is：
8.33mmol/L、6.25mmol/L、4.167mmol/L、3.15mmol/L.Fig. 4 show suction of the different solvents dosage to microballoon
The influence of attached ability.By researching and analysing and comparing the adsorption capacity of polymer obtained under various concentration to template molecule, really
Fixed optimal dosage.
Solvent capacity is formed with important role to polymer, as the reduction of solvent dosage causes system mobility to drop
Low, viscosity increase, monomer molecule movement is difficult, and polymer critical chain length increases, and wraps up the degree of function monomer and reduces, core aggregation
Polymer particle number declines, and when polymerization easily generates bonding, and the reactant frequently resulted in bonds blocking, that is, becomes tube sealing polymerization
Mode causes elution difficulty and increases, and then influences the adsorbance of microballoon.In 30mL, i.e. camplotheca acuminata alkali concn exists solvent
During 4.167mmol/L, adsorption capacity reaches maximum, and the viscosity of reaction system solvent reaches optimum capacity, and continuing, which increases solvent, holds
Amount, monomer concentration reduce, and crosslinked polymer is spent low, cannot partly be reacted completely, be caused the reduction of adsorption capacity.
Reaction temperature determines：Other reaction conditions are constant, and the present embodiment is anti-with the polymerization that numerous studies of the present invention determine
It selects 50 DEG C, 55 DEG C, 60 DEG C to be illustrated as representative in 50 DEG C~60 DEG C of the preferable temperature range answered, investigates different temperatures
Under influence to Macroscopic single crystal, be shown in Table 3.
Influence of 3 synthesis temperature of table to the character of microballoon
When polymerization temperature is 60 DEG C, the absorption property of the MIPs of preparation is best, and character is preferably also.Synthesis temperature determines
Reaction system viscosity size when temperature is more than 60 DEG C, causes polymerization speed to be accelerated, polymer chain is elongated, microsphere mass transfer
Become difficult, while volatile solvent easily escapes in system, mobility is reduced, the microballoon caking of generation；Temperature is too low, generation
Polymer rigid it is too small, be unable to maintain that opening structure, and react incomplete, and the reactant formed under too low synthesis temperature
A kind of state of jello is presented in mixture.
The dosage of function monomer determines：In the case where other conditions are constant, CPT concentration is 4.165mmol/L, changes mould
The ratio of plate molecule and function monomer, the present embodiment is with ratio 1:2、1:4、1:6、1:8 illustrate.Fig. 5 show function list
Influence situation of the body dosage to the absorption property of microballoon.Fig. 6 show the microballoon SEM figures of difference in functionality monomer ratio synthesis.It can
Work as camptothecine to see：MAA molar ratios are 1:When 4, adsorbance reaches maximum.The amount of increase MAA is conducive to itself and camptothecine shape
Into the polymer with hole trace, when molar ratio is 1:When 4, the hole that can be combined in function monomer with template molecule just reaches
To saturation, the association of MAA itself can only be increased by continuing increase, influence the absorption property of microballoon.In general, in dispersion
The monomer of low concentration is more advantageous to being formed the microballoon of grain size bigger[67-68].When monomer concentration is relatively low, polymerization initial stage is formed
Less oligomer core, therefore more free monomers diffuse to the surface of core with crosslinking agent, gradually form the microballoon of bigger.By
SEM electron microscopes can find that monomer concentration increases, and microspherulite diameter is smaller.
The dosage of crosslinking agent determines：It keeps other conditions constant, changes the ratio of template molecule and dosage of crosslinking agent, this reality
Example is applied with 1:10、1:15、1:20、1:It is illustrated exemplified by 30, the concentration (w/v, g/l) for being corresponding in turn to crosslinking agent is：8.26%th,
12.39%th, 16.52%, 24.77%.Fig. 7 show influence situation of the dosage of crosslinking agent to microballoon absorption property.Work as crosslinking
The absorption property for the molecularly imprinted polymer that agent dosage obtains when too high or too low is not ideal, and experiment obtains working as function monomer
It is 1 with crosslinker ratio:When 15, the absorption property of microballoon is optimal.Influence of the degree of cross linking to polymer beads Mesoporous property is close
Related cross-links agent dosage is too small, and the viscosity of polymerization system is reduced, and crosslinking points tail off, and influence the knot of template molecule and function monomer
It closes, while in elution process, macromolecule aggregation state easily adjusts, and makes poroid being destroyed of polymerization stage formation, destroys
The binding site with template molecule that originally forms influences the absorption property of microballoon.Dosage of crosslinking agent is excessive, although formed
Polymer orifices are relatively stablized, but aperture becomes smaller, and influences mass transfer of the template molecule to polymeric inner, cause absorption property under
Drop.
The absorption property of 2 microsphere microballoon (MIPs) of embodiment compares
The microsphere microballoon (MIPs) and blank microsphere (NIPs) prepared with the present invention is under various concentration to mesh
The adsorption capacity of molecule camptothecine is marked, draws the adsorption isotherm of polymer, as shown in Figure 8.As can be seen that the absorption of polymer
Capacity increases with the increase of camplotheca acuminata alkali concn, finally tends to balance.When concentration is relatively low, the adsorption site of polymer is not satisfied also
With, polymer to the target molecule in solution also there is adsorption capacity, and when concentration gradually increases, a certain amount of microsphere gathers
The adsorption capacity for closing object reaches saturation, and adsorption capacity is not further added by and tends to saturation state.
Imprinted polymer is compared with the adsorption capacity of non-imprinted polymer big simultaneously, it was demonstrated that the imprinted polymer of synthesis is to template
Molecule has higher affinity and specific recognition.
Gas selectivity α, microsphere β are introduced to compare the selectivity of microsphere and non-microsphere：
In formula (II)：Cp:During adsorption equilibrium in polymer camptothecine concentration (μm ol/g)；
Cs:During adsorption equilibrium in solution camptothecine concentration (mmol/L).
In formula (III), (IV), Kd,M:The equilibrium dissociation constant of imprinted polymer；
Kd,N:The equilibrium dissociation constant of non-imprinted polymer.
Wherein, separation factor α represents selectivity of the microsphere to template molecule and its analogue, be generally separated because
Sub- α>1.5, that is, think that the microsphere polymer has template molecule special selection identity.Microsphere β representatives are being detained
After non-selection absorption, microsphere polymer is to the ability of template molecule, general microsphere β>0.5, that is, think MIPs ratios
NIPs has significant difference to template molecule, and imprinting effect is good.As shown in table 2, table 3, it can be seen that when solution concentration (0.5~
1.5) in the range of mmol/L, microsphere β is all higher than 0.5, MIPs has significant otherness than NIPs, has better choice
Recognition capability.
The imprinting factor of MIPs and NIPs under 4 various concentration of table
Scatchard analyses are carried out to the absorption property of polymer, equation is：
In formula (V), C be template molecule equilibrium concentration, QmaxFor maximum apparent adsorption quantity.
Fig. 9 show Scatchard analysis charts of the MIPs and NIPs to the absorption property of camplotheca acuminata aqueous slkali.By left in Fig. 9
Figure understands that for Q/C and Q in non-linear relation, this is very universal in non-covalent type molecularly imprinted polymer, illustrates MIPs microballoons pair
Template molecule is anisotropic there are two kinds of different adsorption sites.The reason for generating two kinds of different binding sites may be
Function monomer MAA and template molecule camptothecine are combined with different proportionings during self assembly, form two
Kind imprinted cavity of different nature.It is acquired by the slope and intercept of seeking two straight lines, equilibrium dissociation constant is respectively：
Kd1=1.306mmol/l, Kd2=0.323mmol/L；
Maximum apparent adsorption quantity is：Qmax1=120.22 μm of ol/g, Qmax2=71.65 μm of ol/g.
Rather than the scatchard analysis charts of imprinted polymer are linearly distributed, i.e., its adsorption site is isotropism, is reacted
A kind of binding site has been only formed in the process, that is to say, that NIPs is in non-selective adsorption to the adsorptivity of template molecule.
The Selective adsorption experiment of 3 microsphere microballoon (MIPs) of embodiment
Both experiment uses hydroxycamptothecin the making choice property adsorption experiment similar with camptothecine structure, by comparing
Adsorption capacity size investigates the specific selection adsorptivity of microballoon.
5 MIPs of table compares the absorption property of camptothecine, hydroxycamptothecin
From upper table 5, MIPs is to the adsorbance of target molecule camptothecine apparently higher than the adsorbance to hydroxycamptothecin.
Separation factor α is 1.83~2.09, is all higher than 1.5, shows to select recognition capability well.This is because polymer MIPs exists
Form the stereochemical structure trace hole to match with template molecule (camptothecine) size, form during synthesis, and NIPs simply by
Function monomer is formed with crosslinking agent reactive polymeric, and formation is random, the smaller particle of grain size, does not identify position specifically
Point.It can prove that MIPs produced by the present invention has camptothecine special Selective adsorption.
Claims (10)
1. a kind of preparation method of camptothecine microballoon, which is characterized in that be using the mixed solvent of methanol and chloroform as pore-foaming agent,
MAA is function monomer, and camptothecine is template molecule, and EGDMA is crosslinking agent, under the action of initiator, vibrates and polymerize through water-bath
It is obtained after dry afterwards, elution.
2. the preparation method of camptothecine microballoon according to claim 1, which is characterized in that be by controlling in the mixed solvent first
The amount of alcohol controls the grain size of microballoon；It is preferred that the volume ratio of the in the mixed solvent, methanol and chloroform is 5~8:5~2.
3. the preparation method of camptothecine microballoon according to claim 1, which is characterized in that the dosage of the mixed solvent according to
The gauge of 1moL template molecules, the solvent of addition is 15~40mL；The solvent being preferably added to is 30mL；The temperature of the polymerization
For 50 DEG C~60 DEG C, preferably 60 DEG C；When the time of the polymerization is 18 small.
4. the preparation method of camptothecine microballoon according to claim 1, which is characterized in that the dosage of the function monomer according to
The mol ratio of itself and template molecule is 2~8:1 determines；The dosage of the crosslinking agent is according to its mol ratio with template molecule
For 10~30:1 determines；It is preferred that the mol ratio of the template molecule, function monomer, crosslinking agent is 1:5:15.
5. the preparation method of camptothecine microballoon according to claim 1, which is characterized in that the initiator is different using azo two
Butyronitrile or azo diamyl cyanogen.
6. according to the preparation method of any one of claim 1 to the 5 camptothecine microballoon, which is characterized in that comprise the following steps：
S1. weigh a certain amount of template molecule camptothecine and function monomer MAA is dissolved in a certain amount of solvent, rocked at room temperature is pre-
Polymerization, is completely dissolved camptothecine and forms stable hydrogen bond with MAA；
S2. crosslinking agent EGDMA and initiator A IBN is added in into the system after step S1 prepolymerizations, leads to N2 deoxidations, seal after
Water bath with thermostatic control vibration polymerization under certain temperature；
S3. polymerizate taking-up is cooled to room temperature after the completion of the polymerisation described in step S2, centrifugal drying obtains polymer；
S4. eluted template molecule and unreacted substance, are washed until it can't detect template molecule in eluent, then with solvent
Excessive acetic acid is removed, is dried in vacuo to get to camptothecine microballoon.
7. the preparation method of camptothecine microballoon according to claim 8, which is characterized in that solvent described in step S1 is molten to mix
It is 5~8 that agent, which is methanol and chloroform, according to volume ratio:5~2 ratio-dependent；When the prepolymerized time described in step S1 is 1 small；
The temperature of water bath with thermostatic control described in step S2 is 50 DEG C~60 DEG C；When the time polymerizeing described in step S2 is 18 small.
8. the preparation method of camptothecine microballoon according to claim 8, which is characterized in that lead to N2 deoxidations described in step S2
Time is 10 min；Elution described in step S4 is using V (methanol):V (acetic acid)=9:1 mixed solution uses soxhlet extraction method
Wash off template molecule and unreacted substance；Solvent described in step S4 is methanol.
9. the camptothecine microballoon that the preparation method of any one of claim 1 to the 8 camptothecine microballoon is prepared.
10. application of the camptothecine microballoon as camptothecine imprinted polymer in terms of separating-purifying camptothecine described in claim 9.
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