CN109021173A - A kind of preparation method of thymopeptide-5 molecular engram core-shell particles - Google Patents

A kind of preparation method of thymopeptide-5 molecular engram core-shell particles Download PDF

Info

Publication number
CN109021173A
CN109021173A CN201810641436.7A CN201810641436A CN109021173A CN 109021173 A CN109021173 A CN 109021173A CN 201810641436 A CN201810641436 A CN 201810641436A CN 109021173 A CN109021173 A CN 109021173A
Authority
CN
China
Prior art keywords
solution
core
shell particles
thymopeptide
hevim
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
Application number
CN201810641436.7A
Other languages
Chinese (zh)
Other versions
CN109021173B (en
Inventor
管萍
王姗
胡小玲
李帮鹏
陈婷婷
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Northwestern Polytechnical University
Original Assignee
Northwestern Polytechnical University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Northwestern Polytechnical University filed Critical Northwestern Polytechnical University
Priority to CN201810641436.7A priority Critical patent/CN109021173B/en
Publication of CN109021173A publication Critical patent/CN109021173A/en
Application granted granted Critical
Publication of CN109021173B publication Critical patent/CN109021173B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F267/00—Macromolecular compounds obtained by polymerising monomers on to polymers of unsaturated polycarboxylic acids or derivatives thereof as defined in group C08F22/00
    • C08F267/06—Macromolecular compounds obtained by polymerising monomers on to polymers of unsaturated polycarboxylic acids or derivatives thereof as defined in group C08F22/00 on to polymers of esters
    • 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
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
    • B01J20/26—Synthetic macromolecular compounds
    • B01J20/261—Synthetic macromolecular compounds obtained by reactions only involving carbon to carbon unsaturated bonds
    • 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
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
    • B01J20/26—Synthetic macromolecular compounds
    • B01J20/268—Polymers created by use of a template, e.g. molecularly imprinted polymers
    • 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
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
    • B01J20/28016—Particle form
    • B01J20/28019—Spherical, ellipsoidal or cylindrical
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
    • C08J9/28—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof by elimination of a liquid phase from a macromolecular composition or article, e.g. drying of coagulum
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2201/00—Foams characterised by the foaming process
    • C08J2201/04—Foams characterised by the foaming process characterised by the elimination of a liquid or solid component, e.g. precipitation, leaching out, evaporation
    • C08J2201/05—Elimination by evaporation or heat degradation of a liquid phase
    • C08J2201/0502—Elimination by evaporation or heat degradation of a liquid phase the liquid phase being organic
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J2351/00—Characterised by the use of graft polymers in which the grafted component is obtained by reactions only involving carbon-to-carbon unsaturated bonds; Derivatives of such polymers

Landscapes

  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Polymers & Plastics (AREA)
  • Engineering & Computer Science (AREA)
  • Materials Engineering (AREA)
  • Manufacturing Of Micro-Capsules (AREA)
  • Medicinal Preparation (AREA)

Abstract

The present invention relates to a kind of preparation methods of thymopeptide-5 molecular engram core-shell particles, with the preparation method for the thymopeptide-5 molecular engram core-shell particles that 1- (2- ethoxy) -3- vinyl imidazole chlorine ([HeVIM] Cl) is function monomer, it take 1- (2- ethoxy) -3- vinyl imidazole chlorine ([HeVIM] Cl) as the thymopeptide-5 molecular engram core-shell particles preparation method of function monomer on polyethylene glycol double methyl methacrylate (PEGDMA) caryosphere surface.The preparation method is directed to water-soluble thymopeptide-5 (TP5) molecule, using 1- (2- ethoxy) -3- vinyl imidazole chlorine ([HeVIM] Cl) ionic liquid with a variety of functional groups as function monomer, the water phase trace of TP5 is realized by multiple cooperative interaction and surface imprinted method.Prepared trace core-shell particles have the fast rate of adsorption, higher adsorption capacity and imprinting efficiency, solve the problems, such as biomolecule water phase trace.

Description

A kind of preparation method of thymopeptide-5 molecular engram core-shell particles
Technical field
The invention belongs to the preparation technical fields of functional material, are related to a kind of system of thymopeptide-5 molecular engram core-shell particles Preparation Method is the thymopeptide-5 molecular engram with 1- (2- ethoxy) -3- vinyl imidazole chlorine ([HeVIM] Cl) for function monomer The preparation method of core-shell particles.
Background technique
Polypeptide is related to the multiple function of various cells in organism, to the physiology and pathologic process of people, the generation of disease, Development and treatment etc. play an important role.But polypeptide is normally present in complicated mixture system, and has stability poor, biological The high feature of activity, so that the separation and identification of polypeptide have become a hot topic of research and difficult point.The separation method of traditional polypeptide is for example The disadvantages of extraction, chromatography etc. are long, sample pretreatment requirement is high and at high cost there are extraction time.And surface molecule print method is made For a kind of novel separation and identification technology, prepared molecularly imprinted polymer imprinted sites are mainly distributed on solid-phase matrix table Face has good accessibility, highly beneficial to polypeptide trace.
Thymopeptide-5 (Arg-Lys-Asp-Va1-Tyr, TP5) is artificial synthesized pentapeptide, amino acid sequence and parent The active site 32-36 amino acids residue sequence of thymopoietins is consistent, has bioactivity identical with parent molecule, Immunity can be significantly increased, so being widely used in clinical treatment immunological diseases.
Thymopeptide-5 TP5 is water-soluble biological molecule, and trace need to be usually carried out in water phase, and common is suitable for water phase trace Interference and destruction of the hydrogen bond action that is formed with TP5 such as monomeric acrylic vulnerable to hydrone, reduce imprinting efficiency and accurate Property, so that imprinting effect be made to substantially reduce.And ionic liquid has the characteristics that good designability and stronger solvability, It can be used as reaction dissolvent and function monomer be applied in molecular imprinting technology.Utilize 1- (the 2- hydroxyl second with a variety of functional groups Base) T-MIPs that -3- vinyl imidazole chlorine ([HeVIM] Cl) ionic liquid is prepared as function monomer, multiple collaboration can be passed through Interaction realizes to the good trace of TP5 in aqueous phase system, prepared imprinted polymer will have the faster rate of adsorption, Higher adsorption capacity and imprinting efficiency are expected to solve the problems, such as biomolecule water phase trace.
Currently, there has been no with 1- (2- ethoxy) -3- vinyl imidazole chloride ion liquid ([HeVIM] Cl) be function monomer Thymopeptide-5 molecule (TP5) trace core-shell particles preparation correlative study report.
Summary of the invention
Technical problems to be solved
In order to avoid the shortcomings of the prior art, the present invention proposes a kind of system of thymopeptide-5 molecular engram core-shell particles Preparation Method take 1- (2- ethoxy) -3- vinyl imidazole chlorine ([HeVIM] Cl) as the thymopeptide-5 molecular engram core of function monomer The preparation method of shell microballoon.
Technical solution
A kind of preparation method of thymopeptide-5 molecular engram core-shell particles, it is characterised in that: with 1- (2- ethoxy) -3- second Alkenyl imidazoles chlorine ([HeVIM] Cl) is function monomer, and steps are as follows:
Step 1: by polyvinylpyrrolidone PVP with distilled water mix, stirring is until PVP at 500~700rpm All dissolutions, obtain solution A；The concentration of PVP is 25mg/mL in the solution A；
Step 2: methacrylic acid being added in the mixed solvent, adds azodiisobutyronitrile, bis- (a, the a'- diformazans of S, S'- Guanidine-acetic acid) trithiocarbonate CAT and glycol methacrylate EGDMA, 20~30min of ultrasound obtain clear solution B；The concentration of the methacrylic acid, azodiisobutyronitrile, CAT, EGDMA in solution B be respectively 0.47mmol/mL, 0.013g/mL,0.009g/mL,0.583g/mL；The mixed solvent is methanol and chloroform, ratio V/V=1:7~1:8；
Step 3: under nitrogen atmosphere protection, solution B being added drop-wise in solution A with constant pressure funnel, 500~700rpm Lower stirring, in 50~60 DEG C of condensing reflux 2h；70~80 DEG C will be adjusted in temperature again, reaction is for 24 hours；Resulting product is filtered, And much filtrate is washed 3-5 times with the mixed solution of first alcohol and water, it is dried in vacuo 25~35h at 40~50 DEG C, obtains poly- second two Alcohol double methyl methacrylate PEGDMA caryosphere；The ratio V/V=1:1 of the first alcohol and water；
Step 4: PEGDMA caryosphere being mixed with phosphate buffer solution, sequentially adds TP5, function monomer [HeVIM] Cl And N, N- methylene-bisacrylamide MBA, 25~30min of ultrasonic disperse, it is passed through nitrogen 10~25min deoxygenation, adds 25 μ L N, N- tetramethylethylenediamine TEMED and 45mg ammonium persulfate, the polymerization reaction 10h at 45~50 DEG C filter, it is micro- to obtain nucleocapsid Ball；
The PEGDMA caryosphere and phosphate buffer solution dosage are 4mg/mL；
The ratio between amount of substance of the TP5 and [HeVIM] Cl is 1:45；The ratio between the amount of substance of [HeVIM] Cl and MBA is 1:1~1:4；
The concentration of the phosphate buffer solution is 0.01mol/L, pH=9；
Step 5: core-shell particles methanol and acetic acid mixed solution oscillation elution use UV-vis spectroscopy light after centrifugation Degree meter detects the absorbance of thymopeptide-5 in supernatant at 275.5nm wavelength；If absorbance is higher than 0.001, this step is repeated Suddenly；If absorbance is lower than 0.001, remaining acid and impurity on microballoon is washed with deionized, filters, it is dry in 40~50 DEG C of vacuum Dry 20~30h obtains thymopeptide-5 molecular engram core-shell particles；
The ratio V/V=9:1 of the methanol and acetic acid mixed solution.
Beneficial effect
One kind proposed by the present invention is function monomer with 1- (2- ethoxy) -3- vinyl imidazole chlorine ([HeVIM] Cl) The preparation method of thymopeptide-5 molecular engram core-shell particles, on polyethylene glycol double methyl methacrylate (PEGDMA) caryosphere surface, It take 1- (2- ethoxy) -3- vinyl imidazole chlorine ([HeVIM] Cl) as the thymopeptide-5 molecular engram core-shell particles of function monomer Preparation method.The preparation method is directed to water-soluble thymopeptide-5 (TP5) molecule, utilizes 1- (the 2- hydroxyl with a variety of functional groups Ethyl) -3- vinyl imidazole chlorine ([HeVIM] Cl) ionic liquid as function monomer, passes through multiple cooperative interaction and table Face blotting realizes the water phase trace of TP5.There is prepared trace core-shell particles the fast rate of adsorption, higher absorption to hold Amount and imprinting efficiency, solve the problems, such as biomolecule water phase trace.
There are many bases for 1- (2- ethoxy) -3- vinyl imidazole chlorine ([HeVIM] Cl) function monomer band that the present invention selects Group can be realized the good trace in aqueous phase system to TP5 by multiple cooperative interaction, have the faster rate of adsorption, more Good binding capacity and imprinting efficiency.Research Thinking is provided to the difficulties for solving biomolecule water phase trace and solves way Diameter.
Specific embodiment
Now in conjunction with embodiment, the invention will be further described:
A kind of preparation method of thymopeptide-5 molecular engram core-shell particles, with 1- (2- ethoxy) -3- vinyl imidazole chlorine ([HeVIM] Cl) is the preparation method of the thymopeptide-5 molecular engram core-shell particles of function monomer, it is characterised in that steps are as follows:
The distilled water of a certain amount of polyvinylpyrrolidone (PVP) and 80~100mL is added to round-bottomed flask by step 1 In, the stirring at 500~700rpm obtains solution A until PVP all dissolutions.The concentration of PVP is 25mg/ in the solution A mL。
Methacrylic acid is added in the mixed solvent (V/V=1:7~1:8) being made of methanol and chloroform by step 2, Add azodiisobutyronitrile, bis- (a, the a'- dimethyl acetic acid) trithiocarbonates (CAT) of S, S'- and the double methyl-props of ethylene glycol Olefin(e) acid ester (EGDMA), 20~30min of ultrasound, obtains clear solution B.The methacrylic acid, azodiisobutyronitrile, CAT, Concentration of the EGDMA in solution B is respectively 0.47mmol/mL, 0.013g/mL, 0.009g/mL, 0.583g/mL.
Solution B is slowly dropped in solution A under nitrogen atmosphere protection with constant pressure funnel by step 3,500~ It is stirred under 700rpm, in 50~60 DEG C of condensing reflux 2h.70~80 DEG C will be adjusted in temperature again, reaction is for 24 hours.By resulting product Filter, and wash much filtrate 3-5 times with methanol and water mixed solution (V/V=1:1), at 40~50 DEG C vacuum drying 25~ 35h obtains polyethylene glycol double methyl methacrylate (PEGDMA) caryosphere.
PEGDMA caryosphere and phosphate buffer solution (0.01mol/L, pH=9) are added in round-bottomed flask by step 4, then according to It is secondary to be added a certain amount of TP5, function monomer [HeVIM] Cl and N, N- methylene-bisacrylamide MBA, ultrasonic disperse 25~ 30min is passed through nitrogen 10~25min deoxygenation, adds 25 μ L N, N- tetramethylethylenediamine TEMED and 45mg ammonium persulfates, in Polymerization reaction 10h at 45~50 DEG C filters, obtains core-shell particles.The PEGDMA caryosphere and phosphate buffer solution dosage are 4mg/mL；The ratio between amount of substance of the TP5 and [HeVIM] Cl is 1:45；The ratio between the amount of substance of [HeVIM] Cl and MBA is 1:1~1:4；
Step 5 core-shell particles methanol and acetic acid mixed solution (V/V=9:1) oscillation elution, after centrifugation using it is ultraviolet-can See that spectrophotometer detects the absorbance of thymopeptide-5 in supernatant at 275.5nm wavelength.If absorbance is higher than 0.001, The step is repeated, until absorbance is lower than 0.001.Remaining acid and impurity on microballoon is washed with deionized, filters, in 40~ 50 DEG C of 20~30h of vacuum drying, obtain TP5 molecular engram core-shell particles.
Specific embodiment:
Embodiment 1:
The distilled water of a certain amount of polyvinylpyrrolidone (PVP) and 80mL is added separately to 150mL three-necked flask In, 30min is stirred at 600 rpm, until PVP all dissolves, obtains solution A.The concentration of PVP is 25mg/ in the solution A mL；
Methacrylic acid is added to the mixed solvent (V/V=1:7.5) being made of 1.6mL methanol and 12mL chloroform In, add azodiisobutyronitrile, bis- (a, the a'- dimethyl acetic acid) trithiocarbonates (CAT) of S, S'- and the double methyl of ethylene glycol Acrylate (EGDMA), ultrasonic 20min obtain clear solution B.The methacrylic acid, azodiisobutyronitrile, CAT, EGDMA Concentration in solution B is respectively 0.47mmol/mL, 0.013g/mL, 0.009g/mL, 0.583g/mL；
Under nitrogen atmosphere protection, solution B is slowly dropped in solution A with constant pressure funnel, under 600rpm revolving speed Stirring, in 60 DEG C of condensing reflux 2h.70 DEG C will be adjusted in temperature again, reaction is for 24 hours.Resulting product is filtered, and with methanol and Water (V/V=1:1) mixed solution washs 3 times, and each 100mL is dried in vacuo 10h at 40 DEG C, obtains the double first of polyethylene glycol Base acrylate PEGDMA caryosphere；
PEGDMA caryosphere and 50mL phosphate buffer solution (0.01mol/L, pH=9) are added to 100mL round-bottomed flask, then Sequentially add 0.025mmol TP5,1.125mmol function monomer [HeVIM] Cl and 1.125mmol N, N- methylene bisacrylamide Amide MBA, ultrasonic disperse 30min are passed through nitrogen 10min deoxygenation, and 25 μ L N, N- tetramethylethylenediamine TEMED and 45mg are added Ammonium persulfate, polymerization reaction 10h at 45 DEG C filter, obtain core-shell particles.The PEGDMA caryosphere and phosphate buffer solution dosage For 4mg/mL；The ratio between amount of substance of the TP5 and [HeVIM] Cl is 1:45；The ratio between the amount of substance of [HeVIM] Cl and MBA For 1:1；
Core-shell particles 500mL methanol and acetic acid mixed solution (V/V=9:1) oscillation elution 72h, centrifugation, using it is ultraviolet- Visible spectrophotometer detects the absorbance of thymopeptide-5 in supernatant at 275.5nm wavelength.If absorbance is higher than 0.001, The step is then repeated, until absorbance is lower than 0.001.Remaining acid and impurity on microballoon is washed with deionized, filters, in 45 DEG C vacuum drying 10h, obtain TP5 molecular engram core-shell particles.Through detecting, the adsorption capacity of trace core-shell particles is 13.5mg/ G, imprinting factor 2.12.
Embodiment 2:
The distilled water of a certain amount of polyvinylpyrrolidone (PVP) and 80mL is added separately to 150mL three-necked flask In, 30min is stirred at 600 rpm, until PVP all dissolves, obtains solution A.The concentration of PVP is 25mg/ in the solution A mL；
Methacrylic acid is added to the mixed solvent (V/V=1:7.5) being made of 1.6mL methanol and 12mL chloroform In, add azodiisobutyronitrile, bis- (a, the a'- dimethyl acetic acid) trithiocarbonates (CAT) of S, S'- and the double methyl of ethylene glycol Acrylate (EGDMA), ultrasonic 20min obtain clear solution B.The methacrylic acid, azodiisobutyronitrile, CAT, EGDMA Concentration in solution B is respectively 0.47mmol/mL, 0.013g/mL, 0.009g/mL, 0.583g/mL；
Under nitrogen atmosphere protection, solution B is slowly dropped in solution A with constant pressure funnel, under 600rpm revolving speed Stirring, in 60 DEG C of condensing reflux 2h.70 DEG C will be adjusted in temperature again, reaction is for 24 hours.Resulting product is filtered, and with methanol and Water (V/V=1:1) mixed solution washs 4 times, each 100mL, and 10h is dried in vacuo at 40 DEG C and obtains the double methyl of polyethylene glycol Acrylate PEGDMA caryosphere；
PEGDMA caryosphere and 50mL phosphate buffer solution (0.01mol/L, pH=9) are added to 100mL round-bottomed flask, then Sequentially add 0.025mmol TP5,1.125mmol function monomer [HeVIM] Cl and 2.25mmol N, N- methylene bisacrylamide acyl Amine MBA, ultrasonic disperse 30min are passed through nitrogen 10min deoxygenation, and 25 μ L N, N- tetramethylethylenediamine TEMED and 45mg mistakes are added Ammonium sulfate, polymerization reaction 10h at 45 DEG C filter, obtain core-shell particles.The PEGDMA caryosphere and phosphate buffer solution dosage are 4mg/mL；The ratio between amount of substance of the TP5 and [HeVIM] Cl is 1:45；The ratio between the amount of substance of [HeVIM] Cl and MBA is 1:2；
Core-shell particles 500mL methanol and acetic acid mixed solution (V/V=9:1) oscillation elution 72h, centrifugation, using it is ultraviolet- Visible spectrophotometer detects the absorbance of thymopeptide-5 in supernatant at 275.5nm wavelength.If absorbance is higher than 0.001, The step is then repeated, until absorbance is lower than 0.001.Remaining acid and impurity on microballoon is washed with deionized, filters, in 45 DEG C vacuum drying 10h, obtain TP5 molecular engram core-shell particles.Through detecting, the adsorption capacity of trace core-shell particles is 15.5mg/ G, imprinting factor 2.27.
Embodiment 3:
The distilled water of a certain amount of polyvinylpyrrolidone (PVP) and 80mL is added separately to 150mL three-necked flask In, 30min is stirred at 600 rpm, until PVP all dissolves, obtains solution A.The concentration of PVP is 25mg/ in the solution A mL；
Methacrylic acid is added to the mixed solvent (V/V=1:7.5) being made of 1.6mL methanol and 12mL chloroform In, add azodiisobutyronitrile, bis- (a, the a'- dimethyl acetic acid) trithiocarbonates (CAT) of S, S'- and the double methyl of ethylene glycol Acrylate (EGDMA), ultrasonic 20min obtain clear solution B.The methacrylic acid, azodiisobutyronitrile, CAT, EGDMA Concentration in solution B is respectively 0.47mmol/mL, 0.013g/mL, 0.009g/mL, 0.583g/mL；
Under nitrogen atmosphere protection, solution B is slowly dropped in solution A with constant pressure funnel, under 600rpm revolving speed Stirring, in 60 DEG C of condensing reflux 2h.70 DEG C will be adjusted in temperature again, reaction is for 24 hours.Resulting product is filtered, and with methanol and Water (V/V=1:1) mixed solution washs 5 times, each 100mL, and 10h is dried in vacuo at 40 DEG C and obtains the double methyl of polyethylene glycol Acrylate PEGDMA caryosphere；
PEGDMA caryosphere and 50mL phosphate buffer solution (0.01mol/L, pH=9) are added to 100mL round-bottomed flask, then Sequentially add 0.025mmol TP5,1.125mmol function monomer [HeVIM] Cl and 3.375mmol N, N- methylene bisacrylamide Amide MBA, ultrasonic disperse 30min are passed through nitrogen 10min deoxygenation, and 25 μ L N, N- tetramethylethylenediamine TEMED and 45mg are added Ammonium persulfate, polymerization reaction 10h at 45 DEG C filter, obtain core-shell particles.The PEGDMA caryosphere and phosphate buffer solution dosage For 4mg/mL；The ratio between amount of substance of the TP5 and [HeVIM] Cl is 1:45；The ratio between the amount of substance of [HeVIM] Cl and MBA For 1:3；
Core-shell particles 500mL methanol and acetic acid mixed solution (V/V=9:1) oscillation elution 72h, centrifugation, using it is ultraviolet- Visible spectrophotometer detects the absorbance of thymopeptide-5 in supernatant at 275.5nm wavelength.If absorbance is higher than 0.001, The step is then repeated, until absorbance is lower than 0.001.Remaining acid and impurity on microballoon is washed with deionized, filters, in 45 DEG C vacuum drying 10h, obtain TP5 molecular engram core-shell particles.Through detecting, the adsorption capacity of trace core-shell particles is 20mg/g, Imprinting factor is 2.78.
Embodiment 4:
The distilled water of a certain amount of polyvinylpyrrolidone (PVP) and 80mL is added separately to 150mL three-necked flask In, 30min is stirred at 600 rpm, until PVP all dissolves, obtains solution A.The concentration of PVP is 25mg/ in the solution A mL；
Methacrylic acid is added to the mixed solvent (V/V=1:7.5) being made of 1.6mL methanol and 12mL chloroform In, add azodiisobutyronitrile, bis- (a, the a'- dimethyl acetic acid) trithiocarbonates (CAT) of S, S'- and the double methyl of ethylene glycol Acrylate (EGDMA), ultrasonic 20min obtain clear solution B.The methacrylic acid, azodiisobutyronitrile, CAT, EGDMA Concentration in solution B is respectively 0.47mmol/mL, 0.013g/mL, 0.009g/mL, 0.583g/mL；
Under nitrogen atmosphere protection, solution B is slowly dropped in solution A with constant pressure funnel, under 600rpm revolving speed Stirring, in 60 DEG C of condensing reflux 2h.70 DEG C will be adjusted in temperature again, reaction is for 24 hours.Resulting product is filtered, and with methanol and Water (V/V=1:1) mixed solution washs 5 times, each 100mL, and 10h is dried in vacuo at 40 DEG C and obtains the double methyl of polyethylene glycol Acrylate PEGDMA caryosphere；
PEGDMA caryosphere and 50mL phosphate buffer solution (0.01mol/L, pH=9) are added to 100mL round-bottomed flask, then Sequentially add 0.025mmol TP5,1.125mmol function monomer [HeVIM] Cl and 4.5mmol N, N- methylene bisacrylamide acyl Amine MBA, ultrasonic disperse 30min are passed through nitrogen 10min deoxygenation, and 25 μ L N, N- tetramethylethylenediamine TEMED and 45mg mistakes are added Ammonium sulfate, polymerization reaction 10h at 45 DEG C filter, obtain core-shell particles.The PEGDMA caryosphere and phosphate buffer solution dosage are 4mg/mL；The ratio between amount of substance of the TP5 and [HeVIM] Cl is 1:45；The ratio between the amount of substance of [HeVIM] Cl and MBA is 1:4；
Core-shell particles 500mL methanol and acetic acid mixed solution (V/V=9:1) oscillation elution 72h, centrifugation, using it is ultraviolet- Visible spectrophotometer detects the absorbance of thymopeptide-5 in supernatant at 275.5nm wavelength.If absorbance is higher than 0.001, The step is then repeated, until absorbance is lower than 0.001.Remaining acid and impurity on microballoon is washed with deionized, filters, in 45 DEG C vacuum drying 10h, obtain TP5 molecular engram core-shell particles.Through detecting, the adsorption capacity of trace core-shell particles is 16.5mg/ G, imprinting factor 2.43.

Claims (1)

1. a kind of preparation method of thymopeptide-5 molecular engram core-shell particles, it is characterised in that: with 1- (2- ethoxy) -3- ethylene Base imidazoles chlorine ([HeVIM] Cl) is function monomer, and steps are as follows:
Step 1: by polyvinylpyrrolidone PVP with distilled water mix, the stirring at 500~700rpm is until PVP is whole Dissolution, obtains solution A；The concentration of PVP is 25mg/mL in the solution A；
Step 2: methacrylic acid being added in the mixed solvent, adds azodiisobutyronitrile, bis- (a, a'- the dimethyl second of S, S'- Acid) trithiocarbonate CAT and glycol methacrylate EGDMA, 20~30min of ultrasound obtain clear solution B；Institute Stating the concentration of methacrylic acid, azodiisobutyronitrile, CAT, EGDMA in solution B is respectively 0.47mmol/mL, 0.013g/ mL,0.009g/mL,0.583g/mL；The mixed solvent is methanol and chloroform, ratio V/V=1:7~1:8；
Step 3: under nitrogen atmosphere protection, solution B being added drop-wise in solution A with constant pressure funnel, is stirred under 500~700rpm It mixes, in 50~60 DEG C of condensing reflux 2h；70~80 DEG C will be adjusted in temperature again, reaction is for 24 hours；Resulting product is filtered, is used in combination The mixed solution of first alcohol and water washs much filtrate 3-5 times, and 25~35h is dried in vacuo at 40~50 DEG C, and it is double to obtain polyethylene glycol Methacrylate PEGDMA caryosphere；The ratio V/V=1:1 of the first alcohol and water；
Step 4: PEGDMA caryosphere is mixed with phosphate buffer solution, sequentially adds TP5, function monomer [HeVIM] Cl and N, N- methylene-bisacrylamide MBA, 25~30min of ultrasonic disperse, are passed through nitrogen 10~25min deoxygenation, add 25 μ L N, N- Tetramethylethylenediamine TEMED and 45mg ammonium persulfate, the polymerization reaction 10h at 45~50 DEG C filter, obtain core-shell particles；
The PEGDMA caryosphere and phosphate buffer solution dosage are 4mg/mL；
The ratio between amount of substance of the TP5 and [HeVIM] Cl is 1:45；The ratio between amount of substance of [HeVIM] Cl and MBA is 1:1 ~1:4；
The concentration of the phosphate buffer solution is 0.01mol/L, pH=9；
Step 5: core-shell particles methanol and acetic acid mixed solution oscillation elution use ultraviolet-visible spectrophotometer after centrifugation The absorbance of thymopeptide-5 in supernatant is detected at 275.5nm wavelength；If absorbance is higher than 0.001, this step is repeated；If Absorbance is lower than 0.001, and remaining acid and impurity on microballoon is washed with deionized, filters, is dried in vacuo 20 in 40~50 DEG C ~30h obtains thymopeptide-5 molecular engram core-shell particles；
The ratio V/V=9:1 of the methanol and acetic acid mixed solution.
CN201810641436.7A 2018-06-21 2018-06-21 Preparation method of thymopentin molecularly imprinted core-shell microspheres Expired - Fee Related CN109021173B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201810641436.7A CN109021173B (en) 2018-06-21 2018-06-21 Preparation method of thymopentin molecularly imprinted core-shell microspheres

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201810641436.7A CN109021173B (en) 2018-06-21 2018-06-21 Preparation method of thymopentin molecularly imprinted core-shell microspheres

Publications (2)

Publication Number Publication Date
CN109021173A true CN109021173A (en) 2018-12-18
CN109021173B CN109021173B (en) 2020-07-03

Family

ID=64610429

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201810641436.7A Expired - Fee Related CN109021173B (en) 2018-06-21 2018-06-21 Preparation method of thymopentin molecularly imprinted core-shell microspheres

Country Status (1)

Country Link
CN (1) CN109021173B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108997219A (en) * 2018-07-04 2018-12-14 西北工业大学 A kind of 1- (2- ethoxy) -3- vinyl imidazole chloride ion liquid function monomer and preparation method

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103333295A (en) * 2013-06-18 2013-10-02 西北工业大学 Preparation method of thymopentin molecularly-imprinted magnetic microspheres
CN104140501A (en) * 2014-07-22 2014-11-12 中国科学院烟台海岸带研究所 Thermosensitive bisphenol A (BPA) imprinted polymeric microsphere and application thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103333295A (en) * 2013-06-18 2013-10-02 西北工业大学 Preparation method of thymopentin molecularly-imprinted magnetic microspheres
CN104140501A (en) * 2014-07-22 2014-11-12 中国科学院烟台海岸带研究所 Thermosensitive bisphenol A (BPA) imprinted polymeric microsphere and application thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
CHAOLI WANG ET.AL.: "《Separation and purification of thymopentin with molecular imprinting membrane by solid phase extraction disks》", 《JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108997219A (en) * 2018-07-04 2018-12-14 西北工业大学 A kind of 1- (2- ethoxy) -3- vinyl imidazole chloride ion liquid function monomer and preparation method

Also Published As

Publication number Publication date
CN109021173B (en) 2020-07-03

Similar Documents

Publication Publication Date Title
CN103833915B (en) Molecularly imprinted polymer nano particle suitable for pure biological sample and preparation method thereof
CN104861172B (en) A kind of preparation method of the star copolymer with fluorescent effect, pH responses and temperature-responsive with porphyrin as core
CN103570871A (en) Mesoporous molecular sieve SBA-15 composite nano surface imprinted polymer and preparation method thereof
Farzaneh et al. Molecularly imprinted polymer nanoparticles for olanzapine recognition: application for solid phase extraction and sustained release
CN101718041A (en) Fiber base protein molecularly imprinted polymer aquagel and preparation method thereof
CN103497277A (en) Baicalein molecularly imprinted polymer, preparation method and application thereof
Javanbakht et al. Extraction and purification of penicillin G from fermentation broth by water-compatible molecularly imprinted polymers
CN110078934A (en) A kind of preparation method and its utilization of PDA supermolecular gel
Arabzadeh et al. Synthesis and characterization of molecularly imprinted polymers for selective solid-phase extraction of pseudoephedrine
CN101845127B (en) Method for preparing core-shell structured composite nano surface molecular imprinting polymer of tanshinone compound
CN106632529B (en) A kind of shell tetrose monomer separation extracting method based on molecular imprinting technology
CN108864364B (en) Preparation method of L-phenylalanine molecularly imprinted polymer
CN103333295B (en) Preparation method of thymopentin molecularly-imprinted magnetic microspheres
CN109021173A (en) A kind of preparation method of thymopeptide-5 molecular engram core-shell particles
CN103232572A (en) Molecular imprinting polymer for roxarsone detection, and preparation method thereof
CN103599755A (en) Phenylethanolamine A molecularly imprinted membrane, preparation method thereof and applications thereof
CN103224590B (en) Glabridin molecularly imprinted polymer, as well as preparation method and application thereof
CN103304804B (en) A kind of polyethylene glycol-amide-amine-polyamino acid is linear-dendritic block polymkeric substance and preparation method thereof
CN110590998B (en) Synthesis method and application of sulfonamide dimethylpyrimidine molecularly imprinted polymer
Yang et al. Chiral separation of nateglinide and its l enantiomer on a molecularly imprinted polymer-based stationary phase
CN105542066B (en) A kind of preparation method of spiro-pyrans base random copolymer P (SPMA co MAA)
Shen et al. A triple-function zwitterion for preparing water compatible diclofenac imprinted polymers
CN110404584A (en) A kind of 4-methyl umbelliferone molecular engram nanofiber, preparation method and application
CN108084341B (en) Synthesis method and application of crystal violet molecularly imprinted microspheres
CN109851717A (en) A kind of preparation method and application of anionic PSSS/PSA microballoon

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
CF01 Termination of patent right due to non-payment of annual fee
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20200703

Termination date: 20210621