CN107189011A - Hollow molecules imprinted polymer, solid-phase extraction column and its preparation method and application - Google Patents
Hollow molecules imprinted polymer, solid-phase extraction column and its preparation method and application Download PDFInfo
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- CN107189011A CN107189011A CN201611191091.7A CN201611191091A CN107189011A CN 107189011 A CN107189011 A CN 107189011A CN 201611191091 A CN201611191091 A CN 201611191091A CN 107189011 A CN107189011 A CN 107189011A
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- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F222/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by a carboxyl radical and containing at least one other carboxyl radical in the molecule; Salts, anhydrides, esters, amides, imides, or nitriles thereof
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- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/22—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the construction of the column
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- C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
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- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F222/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by a carboxyl radical and containing at least one other carboxyl radical in the molecule; Salts, anhydrides, esters, amides, imides, or nitriles thereof
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Abstract
The extraction column being made the invention provides a kind of hollow molecules imprinted polymer of phenyl boric acid modification and using the polymer, the synthetic method of the polymer is using 3 amino phenyl boric acids as dressing agent, methacrylic acid monomer is chemically modified, using uridine as template, the methacrylic acid of modification is used as function monomer, add matrix, crosslinking agent and initiator and prepare molecular engram material, eluted template is carried out to obtained molecular engram material, and matrix is eroded using hydrofluoric acid, hollow molecules imprinted polymer is made.The affine hollow molecules trace solid phase of boron that the present invention is provided is mainly due to molecular engram is to the selective absorption of template molecule and analogue and most of binding site of hollow molecules trace is distributed in carrier base material surface and hollow intracavitary, improve adsorption efficiency, in addition, boric acid base group can realize reversible absorption and dissociation to the nucleosides material containing syn diol structure, therefore have obvious concentration effect to nucleosides material.
Description
Technical field
The invention belongs to solid phase extraction techniques field, more particularly to a kind of hollow molecules trace solid phase extraction of phenyl boric acid modification
The preparation method and application method of post are taken, and is combined with efficient liquid-phase chromatography method for Selective Separation, enrichment and detection medicine
The content of nucleosides material in product.
Background technology
Molecular imprinting technology refers to prepare using manual method to be had specially on space structure and binding site to specific molecular
The polymer technology of one sexual reaction.Molecularly imprinted polymer (Molecularly imprinted polymers, MIPs)
Be synthesized by template molecule, the interphase interaction of function monomer and crosslinking agent three it is a kind of have three-D space structure
" acceptor ", has unique " memory " function to template molecule, thus show special compatibility, the selectivity of height and
Remarkable molecule distinguishability.Nevertheless, molecularly imprinted polymer still suffers from imprinted sites distribution not in its preparation process
Homogeneous, " embedding " is too deep, the problems such as unfavorable elution, and in order to solve these problems, some new molecular engram materials are mutually secondary
Exhibition is got up.
Hollow molecules trace (Hollow molecularly imprinted polymers, h-MIPs) technology is used as one
Novel molecular engram technology is planted, is on the basis of surface molecule print technology, to pass through chemolysis or the method for etching, removes
Carrier matrix, obtains the polymer with hollow structure, and polymer made from this novel preparation technology makes most of bound site
Point is distributed on carrier base material surface, be conducive to template molecule elution and object in conjunction with effectively solving
The imprinted sites distributing inhomogeneity that occurs in molecular imprinting technology evolution, " embedding " are too deep, the problems such as unfavorable elution, together
When reduce polymeric inner again and there is diffusional resistance, improve imprinted material adsorption separation efficiency, thus be widely used in solid
In terms of mutually extracting.
Boric acid chemical method is can be with c/s-diol class material, such as ucleosides thing in alkaline aqueous solution using boric acid base group
Cis ortho position or meta-hydroxyl contained by matter occur reversible reaction and form cyclic diester, and realize in acid condition reversibly
Dissociation, so as to carry out separation and concentration to the nucleosides containing syn diol structure.The characteristic of this pH switches makes boric acid turn into excellent
Affine identification aglucon.Using specific recognition of the boric acid to syn diol structure, the h-MIPs of boric acid modified, Ke Yishi are prepared
Now to the Selective recognition and separation and concentration of nucleosides material.Also, syn diol structure can in utilization phenyl boric acid and nucleosides
Reverse action, can be with selectivity of the reinforcing material to specific nucleosides.Simultaneously, it is possible to use pH value controls adsorption/desorption process.
Phenyl boric acid modification hollow molecules trace solid-phase extraction column preparation process in particle diameter, dispersiveness, specific surface area and
The optimization of adsorption effect enables before nucleosides material is effectively enriched with and quickly and easily separated with Matrix Solution by material
Put forward condition.In addition, after MCM-48 matrix is removed, a single hole is formed on the surface of polymer, so that hollow molecules are made
Imprinted polymer, it is desirable to which most of adsorption site of hollow molecules imprinted polymer is all located on the surfaces externally and internally of polymer, this
One feature is conducive to improving the mass transfer rates of polymer while will also improve the adsorption capacity of polymer.Therefore, above-mentioned conjunction
Control into condition is key technical problem that this patent is intended to solve.
The content of the invention
It is an object of the invention to provide a kind of hollow molecules imprinted polymer of phenyl boric acid modification, the polymer is included
The preparation and application of solid-phase extraction column, is combined with high-efficiency liquid chromatography method for detecting, can be achieved cis to containing in medicine
Detected while nucleosides material (cytidine, guanosine, uridine and inosine) content of diol structure.
Specifically, the preparation method of the hollow molecules imprinted polymer for the phenyl boric acid modification that the present invention is provided, including it is as follows
Step：
S1：Using 3- amino phenyl boric acid as dressing agent, methacrylic acid monomer is chemically modified so that methacrylic acid
Surface modification has the phenyl boric acid structure to nucleosides with boron affinity interaction, methacrylic acid monomer and 3- the amino phenyl boric acid
The amount of material compares 1：1~3；
Shown in specific course of reaction such as formula (1)；
S2：Using mesopore molecular sieve MCM-48 as matrix, the uridine with syn diol structure is template, table made from S1
Face is modified with the methacrylic acid of phenyl boric acid structure as function monomer, adds crosslinking agent and initiator prepares molecular engram material
Material, eluted template is carried out to obtained molecular engram material, and erodes matrix mesopore molecular sieve MCM-48 using hydrofluoric acid, is made
Obtain hollow molecules imprinted polymer；
Wherein, the ratio between template, function monomer, crosslinking agent, amount of material of initiator are：2.5~4：10：40~100：
0.6~5, the function monomer often added in 1g matrix is 1.5~5mmol.
Shown in specific course of reaction such as formula (2)；
Preferably, the mesopore molecular sieve MCM-48 is using tetraethyl orthosilicate as silicon source, with cetyl trimethyl bromination
Amine is activating agent, synthesizes what is obtained using hydrothermal synthesis method；
Shown in specific course of reaction such as formula (3)；
Preferably, the crosslinking agent is GDMA, and the initiator is azodiisobutyronitrile.
Preferably, S1 detailed process is：It is according to the amount ratio of material with 3- amino phenyl boric acid by methacrylic acid monomer
1:1~3 ratio mixing, is dissolved in acetonitrile, 12~18h is stirred at room temperature, obtaining surface modification has the first of phenyl boric acid structure
Base acrylic acid.
It is highly preferred that S2 detailed process is：Have to surface modification in the methacrylic acid of phenyl boric acid structure and add template
Uridine, stands 4~6h；Sequentially add crosslinking agent GDMA and initiator azodiisobutyronitrile, make template,
The ratio between amount of material of function monomer, crosslinking agent and initiator is：2.5~4： 10：40~100：0.6~5, lead to N210~
30min, 1~2h is stirred at 40~80 DEG C；Afterwards, mesopore molecular sieve MCM-48,15~30min of ultrasound are added into system, is led to
N2Sealing, stirs at 40~70 DEG C and stirs 6~8h at 18~20h of stirring at 2h, 65~80 DEG C, 80~100 DEG C, polymerization is made
Thing microballoon；
After the polymer microballoon suction filtration ethanol prepared is washed, 10h, mistake are etched with the alcohol solution dipping containing HF
After filter washing, polymer microballoon is placed in cable type extractor according, is washed away after template molecule, is put into methanol/acetic acid mixed solvent
Dried in vacuum desiccator to constant weight, obtain hollow molecules imprinted polymer；
The volume ratio of the methanol/acetic acid in the mixed solvent methanol and acetic acid is 10-X：X, X are selected from appointing between 1~3
Meaning numerical value.
Preferably, present invention also offers a kind of hollow molecules imprinted polymer of phenyl boric acid modification, specifically by above-mentioned
Preparation method described in one is made.
Preferably, present invention also offers a kind of hollow molecules trace solid-phase extraction column of phenyl boric acid modification, including it is hollow
Packed layer is filled with solid-phase extraction column, the hollow solid-phase extraction column, the bottom and top of the packed layer are mounted on sieve
Plate；Packed layer institute filling chock material is the hollow molecules imprinted polymer that above-mentioned phenyl boric acid is modified.
It is highly preferred that present invention also offers the hollow molecules trace solid-phase extraction column of phenyl boric acid modification and efficient liquid phase
Chromatographic detection method is combined, and the application of the nucleosides material content containing syn diol structure is detected in medicine.
It is highly preferred that the nucleosides material is cytidine, guanosine, uridine or inosine.
It is highly preferred that the hollow molecules trace solid-phase extraction column and high-efficiency liquid chromatography method for detecting knot of phenyl boric acid modification
Close, the nucleosides material content containing syn diol structure comprises the following steps that in detection medicine：
The hollow molecules trace solid-phase extraction column that the phenyl boric acid is modified is placed on vacuum solid-phase extraction device, is used successively
Solvent methanol and water activate pillar under negative pressure state, and solvent is drained afterwards, standby；
The hollow molecules trace that sample solution to be detected is continued through into the phenyl boric acid modification under negative pressure state is consolidated
Phase extraction column, drains sample solution；
Eluent is done with normal hexane continue through solid-phase extraction column under negative pressure state and eluted, drain eluent, and
Holding continuously drains state；
Eluant, eluent is done under negative pressure state using 1~5% trifluoroacetic acid/acetonitrile mixed solvent and continues through solid-phase extraction column
Analyte is eluted, eluant, eluent, 1~5% trifluoroacetic acid/acetonitrile in the mixed solvent, 1~5% trifluoroacetic acid is drained
Volume ratio with acetonitrile is 10-Y：Y, Y are selected from any number between 1~5；
Detected after eluent is filtered into high performance liquid chromatograph, determine in medicine and contain syn diol structure
Nucleosides material content.
There is the affine hollow molecules trace solid-phase extraction column of boron that the present invention is provided obvious enrichment to imitate to nucleosides material
Really, mainly due to molecular engram to the big of the selective absorption of template molecule and analogue and hollow molecules trace
Part binding site is distributed in carrier base material surface and hollow intracavitary, improves adsorption efficiency, in addition, boric acid base group to containing
The nucleosides material for having syn diol structure can realize reversible absorption and dissociation.The characteristic of this pH switches makes boric acid turn into excellent
Good affine recognition ligand.The collective effect of three of the above effect so that solid-phase extraction column has obvious adsorption effect.
Brief description of the drawings
The hollow molecules print that Fig. 1 is the carrier matrix MCM-48 that the embodiment of the present invention 1 is provided and the boron prepared is affine
The scanning electron microscope (SEM) photograph of mark polymer.
Embodiment
In order that those skilled in the art more fully understand that technical scheme can be practiced, with reference to specific
The invention will be further described for embodiment, but illustrated embodiment is not as a limitation of the invention.
It should be noted that reagent used in following examples is unless otherwise noted, and it is conventional reagent, can be in city
It is commercially available on, the preparation method or detection method of involved related compound, are conventional side unless otherwise noted
Method.
Embodiment 1
A kind of preparation of the affine hollow molecules trace solid-phase extraction column of boron
1.00mmol monomer methacrylic acid and 1.00mmol 3- amino phenyl boric acids are weighed respectively, and the two is mixed equal
It is even, be dissolved in afterwards in 40mL acetonitriles, 12h stirred at room temperature, obtain methacrylic acid surface modification have to nucleosides have boron parent
With the phenyl boric acid structure of effect；
Template uridine is added into above-mentioned phenyl boric acid structure, addition is to stand 4h at 0.25mmol, 0 DEG C；Add
5.00mmol crosslinking agent GDMA, adds initiator azodiisobutyronitrile 0.457mmol, leads to
N210min, stirs 1h at 40 DEG C；Afterwards, 200mg mesopore molecular sieve MCM-48, ultrasonic 15min is added into system, leads to N2It is close
Envelope, stirs to stir at 2h, 65 DEG C at 40 DEG C and stirs 6h at 18h, 80 DEG C, and polymer microballoon is made；
After the above-mentioned polymer microballoon suction filtration ethanol washing prepared, etched with the alcohol solution dipping containing HF
After 6h, filtration washing, polymer microballoon is placed in cable type extractor according, is 9 with volume ratio:1 methanol/acetic acid mixed solvent is washed
Go after template molecule, be put into vacuum desiccator and dry to constant weight, obtain hollow molecules imprinted polymer, the hollow molecules trace
The scanning electron microscope (SEM) photograph of polymer is as shown in Figure 1；
The above-mentioned hollow molecules imprinted polymer materials of 200mg are weighed, fills to hollow solid phase of the 5mL bottoms equipped with sieve plate and extracts
Take in post, then place sieve plate in the top of filler and be compacted, complete.
The application of the above-mentioned affine hollow molecules trace solid-phase extraction column of the boron prepared
Using Ginkgo Damo injection as actual sample, the enrichment and purification of actual sample are carried out, specific step is, smart
It is close to measure actual sample 2mL, put in 50mL volumetric flasks, plus mobile phase is diluted to 50mL scales, shakes up, and takes appropriate solution to tool
Fill in centrifuge tube, in 4000rmin-1Under the conditions of centrifugation 10min after, filtering, take filtrate, and regard this filtrate as actual sample
Load solution；
The affine hollow molecules trace solid-phase extraction column of boron is placed on vacuum solid-phase extraction device, successively with 3 mL first
Alcohol, 3mL water is activated under negative pressure state, and drains activating solvent；
Above-mentioned actual sample load solution 2mL is continued through into solid-phase extraction column under negative pressure state, made in actual sample
Gradient elution in solid-phase extraction column be adsorbed, drain load solution；Eluent is done in negative pressure state with 3mL normal hexanes again
Under continue through solid-phase extraction column and eluted, drain eluent, and keep continuously draining state；Using 0.5mL volume ratios as 7:3
5% trifluoroacetic acid/acetonitrile mixed solvent as eluant, eluent, solid-phase extraction column is continued through under negative pressure state to analyte
Eluted, drain eluant, eluent；Eluent is filtered into laggard high performance liquid chromatography detection.
Comparative example 1
A kind of preparation of the affine hollow non-molecularly imprinted solid phase extraction column of boron, its preparation method and embodiment 1 are identical, different
Part is only that, template molecule uridine is added without during reaction.
Comparative example 2
Using Ginkgo Damo injection as actual sample, directly using high performance liquid chromatography to the nucleosides material in medicine
Detected, its specific analytical method and embodiment 1 are identical.
Comparative example 3
Using Ginkgo Damo injection as actual sample, directly combined using the solid-phase extraction column UF-C18 of commercialization high
Effect liquid phase chromatogram detects that its specific analytical method and embodiment 1 are identical to the nucleosides material in medicine.
Embodiment 2
A kind of preparation of the affine hollow molecules trace solid-phase extraction column of boron
1.00mmol monomer methacrylic acid and 2.00mmol 3- amino phenyl boric acids are weighed respectively, and the two is mixed equal
It is even, be dissolved in afterwards in 50mL acetonitriles, 16h stirred at room temperature, obtain methacrylic acid surface modification have to nucleosides have boron parent
With the phenyl boric acid structure of effect；
Template uridine is added into above-mentioned phenyl boric acid structure, addition is to stand 4h at 0.4mmol, 20 DEG C；Add 4mmol
Crosslinking agent GDMA, add initiator azodiisobutyronitrile 0.061mmol, lead to N220min, 60
2h is stirred at DEG C；Afterwards, 400mg mesopore molecular sieve MCM-48, ultrasonic 20min is added into system, leads to N2Sealing, at 60 DEG C
Lower stirring 3h, stirs at 80 DEG C and stirs 6h at 20h, 100 DEG C, and polymer microballoon is made；
After the above-mentioned polymer microballoon suction filtration ethanol washing prepared, etched with the alcohol solution dipping containing HF
After 10h, filtration washing, polymer microballoon is placed in cable type extractor according, is 8 with volume ratio:2 methanol/acetic acid mixed solvent
Wash away after template molecule, be put into vacuum desiccator and dry to constant weight, obtain hollow molecules imprinted polymer；
The above-mentioned hollow molecules imprinted polymer materials of 200mg are weighed, fills to hollow solid phase of the 5mL bottoms equipped with sieve plate and extracts
Take in post, then place sieve plate in the top of filler and be compacted.
The application of the above-mentioned affine hollow molecules trace solid-phase extraction column of the boron prepared
Using Clary injection as actual sample, the enrichment and purification of actual sample are carried out, specific step is that precision is measured
Actual sample 5mL is taken, is put in 50mL volumetric flasks, plus mobile phase is diluted to 50mL scales, shakes up, and takes appropriate solution to be centrifuged to having plug
Guan Zhong, in 4000rmin-1Under the conditions of after centrifugation 20min, filtering takes filtrate, and this filtrate is molten as actual sample loading
Liquid；
The affine hollow molecules trace solid-phase extraction column of boron is placed on vacuum solid-phase extraction device, successively with 3 mL second
Nitrile, 3mL water is activated under negative pressure state, and drains activating solvent；
Above-mentioned load solution is continued through into solid-phase extraction column under negative pressure state, makes the gradient elution in actual sample
It is adsorbed in solid-phase extraction column, drains load solution；Eluent is done with 5mL acetone again and continues through solid phase under negative pressure state
Extraction column is eluted, and drains eluent, and keeps continuously draining state；Using 1.0mL volume ratios as 6:4 2% trifluoroacetic acid/
The mixed solvent of acetonitrile is as eluant, eluent, and under negative pressure state continue through solid-phase extraction column elutes to analyte, drains
Eluant, eluent；Eluent is filtered into laggard efficient liquid phase detection.
Comparative example 4
A kind of preparation of the affine hollow non-molecularly imprinted solid phase extraction column of boron, its preparation method and embodiment 2 are identical, different
Part is only that, template molecule uridine is added without during reaction.
Comparative example 5
Using Clary injection as actual sample, directly the nucleosides material in medicine is carried out using high performance liquid chromatography
Detection, its specific analytical method and embodiment 2 are identical.
Comparative example 6
Using Clary injection as actual sample, directly using the solid-phase extraction column UF-C18 combination efficient liquid phases of commercialization
Chromatogram detects that its specific analytical method and embodiment 2 are identical to the nucleosides material in medicine.
Embodiment 3
A kind of preparation of the affine hollow molecules trace solid-phase extraction column of boron
1.00mmol monomer methacrylic acid and 3.00mmol 3- amino phenyl boric acids are weighed respectively, and the two is mixed equal
It is even, be dissolved in afterwards in 60mL acetonitriles, 18h stirred at room temperature, obtain methacrylic acid surface modification have to nucleosides have boron parent
With the phenyl boric acid structure of effect；
Template uridine is added into above-mentioned phenyl boric acid structure, addition is to stand 6h at 0.25mmol, 30 DEG C；Add
10mmol crosslinking agent GDMAs, add the mmol of initiator azodiisobutyronitrile 0.12, lead to N230min,
1h is stirred at 80 DEG C；Afterwards, 600mg mesopore molecular sieve MCM-48, ultrasonic 30min is added into system, leads to N2Sealing, 70
Stir to stir at 4h, 80 DEG C at DEG C and stir 8h at 20 h, 90 DEG C, polymer microballoon is made；
After the above-mentioned polymer microballoon suction filtration ethanol washing prepared, etched with the alcohol solution dipping containing HF
After 10h, filtration washing, polymer microballoon is placed in cable type extractor according, is 7 with volume ratio:3 methanol/acetic acid mixed solvent
Wash away after template molecule, be put into vacuum desiccator and dry to constant weight, obtain hollow molecules imprinted polymer；
The above-mentioned hollow molecules imprinted polymer materials of 200mg are weighed, fills to hollow solid phase of the 5mL bottoms equipped with sieve plate and extracts
Take in post, then place sieve plate in the top of filler and be compacted.
The application of the above-mentioned affine hollow molecules trace solid-phase extraction column of the boron prepared
Using safflower oral liquid as actual sample, the enrichment and purification of actual sample are carried out, specific step is that precision is measured
Actual sample 2mL is taken, is put in 50mL volumetric flasks, plus mobile phase is diluted to 50mL scales, shakes up, and takes appropriate solution to be centrifuged to having plug
Guan Zhong, in 4000rmin-1Under the conditions of centrifugation 30min after, filtering, take filtrate；
The affine hollow molecules trace solid-phase extraction column of boron is placed on vacuum solid-phase extraction device, successively with 5 mL second
Nitrile, 5mL water is activated under negative pressure state, and drains activating solvent；
Above-mentioned filtrate is continued through into solid-phase extraction column under negative pressure state, solvent is drained；Eluted again with 1mL normal butanes
Agent continues through solid-phase extraction column under negative pressure state and eluted, and drains eluent, and keeps continuously draining state；With
1.0mL volume ratios are 5:The mixed solvent of 5 1% trifluoroacetic acid/acetonitrile is continued through solid as eluant, eluent under negative pressure state
Phase extraction column is eluted to analyte, drains eluant, eluent；Eluent is filtered into laggard efficient liquid phase detection.
Comparative example 7
A kind of preparation of the affine hollow non-molecularly imprinted solid phase extraction column of boron, its preparation method and embodiment 3 are identical, different
Part is only that, template molecule uridine is added without during reaction.
Comparative example 8
Using safflower oral liquid as actual sample, directly the nucleosides material in medicine is carried out using high performance liquid chromatography
Detection, its specific analytical method and embodiment 3 are identical.
Comparative example 9
Using safflower oral liquid as actual sample, directly using the solid-phase extraction column UF-C18 combination efficient liquid phases of commercialization
Chromatogram detects that its specific analytical method and embodiment 3 are identical to the nucleosides material in medicine.
Detection solution obtained by above-described embodiment 1-3, and the liquid phase testing conditions of comparative example 1-9 detection solution are equal
For：Chromatographic column：Diamonsil C18 (250mm × 4.6mm, 5 μm)；Mobile phase：Water-methanol (V:V=90: 10)；Flow velocity：
0.9mL·min-1；Detection wavelength：260nm；Column temperature：30℃；Sample size： 10μL.The liquid-phase chromatographic analysis result of gained is seen below
Shown in table 1.
Each embodiment liquid-phase chromatographic analysis result of table 1
The affine hollow molecules trace solid-phase extraction column of boron that 1-3 of the embodiment of the present invention is provided prints with the hollow non-molecule of comparative example
Mark solid-phase extraction column, commercialization solid-phase extraction column UF-C18 are compared to the adsorption effect of nucleosides material, are as a result shown, this
Invention provide the affine hollow molecules trace solid-phase extraction column of boron there is obvious concentration effect to nucleosides material, mainly by
In molecular engram to the selective absorption of template molecule and analogue and most of binding site of hollow molecules trace
Carrier base material surface and hollow intracavitary are distributed in, adsorption efficiency is improved, boric acid base group is to containing c/s-diol knot in addition
The nucleosides material of structure can realize reversible absorption and dissociation.The characteristic of this pH switches makes boric acid turn into excellent affine identification
Part.Three of the above effect collective effect so that the h-MIPs solid-phase extraction columns that the present invention is synthesized have obvious adsorption effect.
Embodiment described above is only the preferred embodiment to absolutely prove the present invention and being lifted, and its protection domain is not limited
In this.Equivalent substitute or conversion that those skilled in the art are made on the basis of the present invention, the protection in the present invention
Within the scope of, protection scope of the present invention is defined by claims.
Claims (10)
1. a kind of preparation method of the hollow molecules imprinted polymer of phenyl boric acid modification, it is characterised in that comprise the following steps：
S1：Using 3- amino phenyl boric acid as dressing agent, methacrylic acid monomer is chemically modified so that metering system acid surfaces
It is modified with the phenyl boric acid structure to nucleosides with boron affinity interaction, the material of methacrylic acid monomer and 3- the amino phenyl boric acid
Amount compare 1：1~3；
S2：Using mesopore molecular sieve MCM-48 as matrix, the uridine with syn diol structure is template, surface modification made from S1
There is the methacrylic acid of phenyl boric acid structure as function monomer, add crosslinking agent and initiator prepares molecular engram material, to system
The molecular engram material obtained carries out eluted template, and erodes matrix mesopore molecular sieve MCM-48 using hydrofluoric acid, is made hollow
Molecularly imprinted polymer；
Wherein, the ratio between template, function monomer, crosslinking agent, amount of material of initiator are：2.5~4：10：40~100：0.6~
5, the function monomer often added in 1g matrix is 1.5~5mmol.
2. the preparation method of the hollow molecules imprinted polymer of phenyl boric acid modification according to claim 1, it is characterised in that
The mesopore molecular sieve MCM-48 is, using tetraethyl orthosilicate as silicon source, using CTAB as activating agent, to use
Hydrothermal synthesis method synthesis is obtained.
3. the preparation method of the hollow molecules imprinted polymer of phenyl boric acid modification according to claim 1, it is characterised in that
The crosslinking agent is GDMA, and the initiator is azodiisobutyronitrile.
4. the preparation method of the hollow molecules imprinted polymer of phenyl boric acid modification according to claim 1, it is characterised in that
S1 detailed process is：
According to the amount ratio of material it is 1 by methacrylic acid monomer and 3- amino phenyl boric acid:1~3 ratio mixing, is dissolved in acetonitrile
In, 12~18h is stirred at room temperature, and obtaining surface modification has the methacrylic acid of phenyl boric acid structure.
5. the preparation method of the hollow molecules imprinted polymer of phenyl boric acid modification according to claim 2, it is characterised in that
S2 detailed process is：
There is addition template uridine in the methacrylic acid of phenyl boric acid structure to surface modification, stand 4~6h；Sequentially add crosslinking agent
GDMA and initiator azodiisobutyronitrile, make the material of template, function monomer, crosslinking agent and initiator
The ratio between amount be：2.5~4：10：40~100：0.6~5, lead to N210~30min, 1~2h is stirred at 40~80 DEG C；Afterwards, to
Mesopore molecular sieve MCM-48,15~30min of ultrasound are added in system, leads to N2Sealing, stirs 2h, 65~80 DEG C at 40~70 DEG C
Lower stirring 18~20h, stirs 6~8h at 80~100 DEG C, polymer microballoon is made；
After the polymer microballoon suction filtration ethanol prepared is washed, 10h is etched with the alcohol solution dipping containing HF, diafiltration is crossed
After washing, polymer microballoon is placed in cable type extractor according, is washed away with methanol/acetic acid mixed solvent after template molecule, is put into vacuum
Dried in drier to constant weight, obtain hollow molecules imprinted polymer；
The volume ratio of the methanol/acetic acid in the mixed solvent methanol and acetic acid is 10-X：X, X are selected from the Arbitrary Digit between 1~3
Value.
6. a kind of hollow molecules imprinted polymer of phenyl boric acid modification, it is characterised in that as described in claim any one of 1-5
Preparation method is made.
7. a kind of hollow molecules trace solid-phase extraction column of phenyl boric acid modification, it is characterised in that including hollow solid-phase extraction column, institute
State and packed layer is filled with hollow solid-phase extraction column, the bottom and top of the packed layer are mounted on sieve plate；
Packed layer institute filling chock material is the hollow molecules imprinted polymer of the phenyl boric acid modification described in claim 6.
8. the hollow molecules trace solid-phase extraction column of phenyl boric acid according to claim 7 modification, it is characterised in that with it is efficient
Liquid chromatography detecting method is combined, and the application of the nucleosides material content containing syn diol structure is detected in medicine.
9. application according to claim 8, it is characterised in that the nucleosides material is cytidine, guanosine, uridine or flesh
Glycosides.
10. application according to claim 8, it is characterised in that comprise the following steps that：
The hollow molecules trace solid-phase extraction column that the phenyl boric acid is modified is placed on vacuum solid-phase extraction device, and solvent is used successively
First alcohol and water activates pillar under negative pressure state, and solvent is drained afterwards, standby；
Sample solution to be detected is continued through to the hollow molecules trace solid phase extraction of the phenyl boric acid modification under negative pressure state
Post is taken, sample solution is drained；
Eluent is done with normal hexane continue through solid-phase extraction column under negative pressure state and eluted, drain eluent, and keep
Continuously drain state；
Eluant, eluent is done using 1~5% trifluoroacetic acid/acetonitrile mixed solvent solid-phase extraction column is continued through under negative pressure state to dividing
Analysis thing is eluted, and drains eluant, eluent, 1~5% trifluoroacetic acid/acetonitrile in the mixed solvent, 1~5% trifluoroacetic acid and second
The volume ratio of nitrile is 10-Y：Y, Y are selected from any number between 1~5；
Detected after eluent is filtered into high performance liquid chromatograph, determine the nucleosides containing syn diol structure in medicine
Class content of material.
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