CN111220745A - Chromatographic column and preparation method thereof - Google Patents
Chromatographic column and preparation method thereof Download PDFInfo
- Publication number
- CN111220745A CN111220745A CN201811417535.3A CN201811417535A CN111220745A CN 111220745 A CN111220745 A CN 111220745A CN 201811417535 A CN201811417535 A CN 201811417535A CN 111220745 A CN111220745 A CN 111220745A
- Authority
- CN
- China
- Prior art keywords
- liquid crystal
- polymer particles
- column
- porous polymer
- internal structure
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/60—Construction of the column
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The invention discloses a chromatographic column applied to gel permeation chromatography, which comprises a hollow column tube and a stationary phase distributed in the column tube, wherein the stationary phase is formed by stacking porous polymer particles, and the porous polymer particles have a regular internal structure and pore channel arrangement and are basically spherical when being swelled. The invention also discloses a method for preparing the chromatographic column, which comprises the following steps: preparing porous polymer particles with a regular internal structure and pore channel arrangement; dispersing porous polymer particles in the homogenate for swelling to form a suspension; the suspension was packed into an empty column tube. The chromatographic column can effectively shorten the separation time of a sample, save a mobile phase and improve the separation effect under the condition of ensuring the separation range.
Description
Technical Field
The invention relates to a chromatographic column, in particular to a gel chromatographic column taking porous polymer microspheres with regular internal structures and pore canal arrangements as a stationary phase and a preparation method thereof.
Background
Gel Permeation Chromatography (GPC), which is a branch of Size Exclusion Chromatography (SEC), is mainly used for the separation of polymers and the molecular weight determination of polymers. The fundamental principle of GPC is that when a sample containing polymer molecules of different sizes passes through a chromatographic column containing porous gel packing, small molecules can diffuse into the gel through smaller and larger pores, larger molecules can only enter the larger pores, and macromolecules larger than the largest pores cannot enter the pores, and can only stay in the gaps between the packing particles and flow out of the chromatographic column together with a mobile phase. With the continuous elution of the mobile phase, the larger molecules are eluted first, and the smaller molecules are eluted later, thereby realizing the separation of molecules with different sizes. Wherein, the lgM-V calibration curve drawn by the relation of the relative molecular weight and the elution volume determines the characteristics of the gel chromatographic column, and the relative molecular mass range of the two ends is called the fractionation range of the gel chromatographic column.
The gel permeation chromatography uses organic solvent as mobile phase, and the commonly used mobile phase is tetrahydrofuran, chloroform, dimethylformamide and the like. Since the mobile phase only acts to dissolve the sample and does not control the degree of separation, the separation of the column is independent of the interaction between the sample and the mobile phase. In contrast, the stationary phase, which is a determining factor for determining the separation effect, must have the following conditions: the medium used as the stationary phase must be an inert substance, and cannot have any effect with the sample to be separated and the solvent in the separation process; the pore diameter and the particle size distribution of the stationary phase are uniform; the stationary phase has excellent physical and chemical stability and high mechanical strength. The stationary phase of the gel permeation chromatography is porous styrene-divinylbenzene copolymer microspheres (PS-DVB) which have uniform particle size, high mechanical strength, pressure resistance and acid and alkali resistance. However, the pore size distribution range of the microspheres is large, the pore channels in the microspheres are irregularly distributed, and the molecules are diffused in the pore channels to have non-uniformity, so that the final separation speed and the final separation effect are influenced.
Therefore, it is required to provide a gel permeation chromatography column, the stationary phase of which has a regular porous internal structure and regularly distributed internal pore channels, so as to improve the separation speed and separation effect.
Disclosure of Invention
In order to meet the above requirements, the present invention provides a chromatography column for gel permeation chromatography, comprising a hollow column tube and a stationary phase distributed in the column tube, wherein the stationary phase is formed by stacking porous polymer particles, the porous polymer particles have a regular internal structure and pore channel arrangement, and the porous polymer particles are substantially spherical when swelling.
In an embodiment of the present invention, the material of the porous polymer fine particles is an acrylic polymer.
In some embodiments of the invention, the regular internal structure and pore channel arrangement of the porous polymeric microparticles has a radial conformation. In other embodiments of the present invention, the regular internal structure and pore channel arrangement of the porous polymeric microparticles has a bipolar conformation.
In an embodiment of the invention, the porous polymeric microparticles have an average particle size when dry in the range of 5 to 100 microns. In a preferred embodiment, the porous polymeric microparticles have an average particle size when dry in the range of 5 to 60 microns. In a further preferred embodiment, the porous polymeric microparticles have an average particle size when dry in the range of 5 to 30 microns.
In an embodiment of the invention, the fractionation range of the chromatography column is 300 to 100000. In a preferred embodiment, the fractionation range of the column is 500 to 50000.
The invention also discloses a method for preparing a chromatographic column applied to gel permeation chromatography, which comprises the following steps: preparing porous polymer particles with a regular internal structure and pore channel arrangement; dispersing the porous polymer particles into the homogenate for swelling to form a suspension; the suspension was packed into an empty column tube. In an embodiment of the invention, the homogenate is tetrahydrofuran, chloroform, toluene or dimethylformyl.
In an embodiment of the invention, the suspension is packed into the hollow column tube using a homogenization method.
In a preferred embodiment, the step of preparing porous polymeric microparticles having a regular internal structure and pore channel arrangement further comprises: forming a homogeneous liquid crystal mixture, wherein the liquid crystal mixture comprises reactive liquid crystals, non-reactive liquid crystals and a polymerization initiator; emulsifying the liquid crystal mixture to form liquid crystal microdroplets dispersed in a continuous phase containing a liquid crystal conformation change agent, wherein the liquid crystal conformation change agent can ensure that liquid crystal molecules in the liquid crystal microdroplets are regularly arranged; polymerizing reactive liquid crystals in the liquid crystal microdroplets to form intermediate microspheres; removing the non-polymerized non-reactive liquid crystal from the intermediate microspheres to form porous polymer microparticles having a regular internal structure and pore channel arrangement.
In an embodiment of the invention, the reactive liquid crystal is present in an amount of 25% to 50% by weight of the total liquid crystal mixture.
In embodiments of the invention, the liquid crystal conformation change agent is a surfactant or a salt.
The chromatographic column applied to the gel permeation chromatography disclosed by the invention utilizes the stacking of the porous polymer particles with regular internal structures and pore canal arrangement to form the stationary phase, and can effectively reduce mass transfer resistance, shorten the separation time of a sample, save a mobile phase and improve the separation effect under the condition of ensuring the separation range.
Drawings
The invention may be better understood by reference to the following description of embodiments of the invention taken in conjunction with the accompanying drawings, in which:
FIG. 1 is a schematic diagram of the structure of a chromatography column disclosed herein;
FIG. 2 is a schematic structural view of porous polymeric microparticles made according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of a membrane emulsification device for preparing liquid crystal droplets;
FIG. 4 is (a) parallel and (b) cross-polarized microscopy images (same scale for multiple microscopy images) of porous polymer microparticles prepared according to embodiments of the invention when dry;
FIG. 5 is a log M-V calibration curve for a chromatography column prepared in accordance with an embodiment of the present invention;
FIG. 6 is a chromatogram of a column separation mixed sample prepared according to an embodiment of the present invention: (a) the chromatography column of example 1, (b) a contrast column;
FIG. 7 is a parallel and cross polarization microscope photograph of porous polymer particles prepared according to an example of the present invention in (a) dried and (b) tetrahydrofuran, and (c) a surface SEM photograph (scale of multiple microscope photographs is the same);
FIG. 8 is a log M-V calibration curve for a chromatography column prepared in accordance with an embodiment of the present invention;
FIG. 9 is (a) parallel and (b) cross-polarized microscopy images (same scale for multiple microscopy images) of porous polymer microparticles prepared according to embodiments of the invention when dry;
FIG. 10 is a log M-V calibration curve for a chromatography column prepared in accordance with an embodiment of the present invention;
FIG. 11 is (a) parallel and (b) cross-polarized microscopy images (same scale for multiple microscopy images) of porous polymer microparticles prepared according to embodiments of the invention when dry;
FIG. 12 is (a) parallel and (b) cross-polarized microscopy images (same scale for multiple microscopy images) of porous polymer microparticles prepared according to embodiments of the invention when dry;
FIG. 13 is a log M-V calibration curve for a chromatography column prepared according to an embodiment of the invention.
Detailed Description
In the following description, for purposes of explanation, numerous specific details are set forth in order to provide a thorough understanding of the present invention, however, it will be apparent to those skilled in the art that the present invention may be practiced without these specific details. In other instances, well-known structures and devices are shown in block diagram form. In this regard, the illustrated example embodiments are provided for purposes of illustration only and are not intended to be limiting of the invention. Therefore, it is intended that the scope of the invention be limited not by this detailed description, but rather by the claims appended hereto.
Abbreviations and molecular formulas as used herein are listed:
5 CB: 4-cyano-4' -pentylbiphenyl
RM 257: 2-methyl-1, 4-phenylene-bis [4- (3-acryloyloxypropoxy) benzoate ]
DMPAP: 2-bis-methoxy-2-phenyl ethanone
SPG film: shirasu Porous Glass membrane
SDS (sodium dodecyl sulfate): sodium dodecyl sulfate
PVP: polyvinylpyrrolidone
PVA: polyvinyl alcohol
PEG: polyethylene glycol
Pluronic: polyoxyethylene polyoxypropylene ether block copolymer
As shown in FIG. 1, the present invention discloses a chromatography column applied to a gel permeation chromatography column, comprising a hollow column tube 11 and a stationary phase distributed therein, wherein the stationary phase is formed by stacking porous polymer particles 12, wherein the porous polymer particles 12 can swell in a good solvent and are substantially spherical when swollen. The porous polymer particles 12 have an average particle size in the range of 5 to 100 micrometers when dry, preferably, the porous polymer particles 12 have an average particle size in the range of 5 to 60 micrometers when dry; more preferably, the porous polymeric microparticles 12 have an average particle size in the range of 5 to 30 microns when dry. The column tube adopts the materials and the sizes which are commonly used for gel chromatographic columns.
The porous polymer particles 12 have pore arrangements with different pore sizes, and when molecules 101 with different sizes (molecular weights) pass through a chromatographic column containing the porous polymer particles 12, smaller molecules can diffuse into the porous polymer particles 12 through the smaller pore passages, larger molecules can only enter the larger pore passages, but macromolecules larger than the pore passages cannot enter the inner pore passages of the porous polymer particles 12, and only stay in the particle gaps of the porous polymer particles 12, flow out of the chromatographic column together with a mobile phase, and leave the chromatographic column at first, so that the separation of the molecules with different sizes is realized. Meanwhile, since the porous polymer particles 12 have a regular internal structure and arrangement of pore channels, the diffusion paths of the molecules 101 of the same size into and out of the porous polymer particles 12 are regular and definite. Compared with the stationary phase with irregularly arranged inner pore channels, the speed of the sample molecules to be separated leaving the chromatographic column is faster, namely the retention time is further shortened. The regular internal structure and the pore arrangement of the porous polymer microparticles 12 may have any regular conformation, and in a ray-type conformation (as shown in fig. 2 (a)), the pores are arranged in the radial direction, thereby exhibiting optical anisotropy of ray type (maltese black cross) under a polarization microscope; in the bipolar conformation (as shown in fig. 2 (b)), the channels are parallel to the circumferential tangent and parallel to the bipolar axis (dashed line in the figure), thus showing the optical anisotropy of the bipolar under a polarization microscope. However, the invention is not limited thereto and other regular conformations may be used.
The range of fractionation of the chromatographic columns prepared therefrom may vary depending on the particle size, pore size and pore distribution of the porous polymeric microparticles 12. In an embodiment of the invention, the fractionation range of the chromatography column is between 300 and 100000. In a preferred embodiment of the invention, the fractionation range of the chromatography column is between 500 and 50000.
The invention also provides a method for preparing a chromatographic column applied to gel permeation chromatography, which comprises the following steps.
In a first step, porous polymer particles having a regular internal structure and pore channel arrangement are prepared. The porous polymer particles can be prepared by a liquid crystal-assisted template polymerization process, which comprises the steps of:
first, a reactive liquid crystal, a non-reactive liquid crystal and a polymerization initiator are mixed in a certain ratio to form a uniform liquid crystal mixture. Wherein the reactive liquid crystal has a polymerizable chemical group, and can react in the presence of a polymerization initiator to form a polymer, such as an acrylate liquid crystal (RM257), a methacrylate liquid crystal (HCM062), an allylic liquid crystal (HCM126), and the like. The mass percentage of the reactive liquid crystal in the liquid crystal mixture can be adjusted between 25% and 50%. The non-reactive liquid crystal has no polymerizable chemical groups and does not polymerize further. The non-reactive liquid crystal comprises at least one nematic liquid crystal, such as the commonly used nematic liquid crystal 5CB or nematic mixed crystal E7.
The liquid crystal mixture is then emulsified to form liquid crystal droplets dispersed in the continuous phase, wherein the liquid crystal droplets comprise a reactive liquid crystal, a non-reactive liquid crystal, and a polymerization initiator. In an embodiment of the invention, the continuous phase is water. The method of emulsification may include stirring, shaking, ultrasonic method, membrane emulsification and the like. In the embodiment of the invention, in order to better control the particle size and distribution of the liquid crystal droplets, the liquid crystal mixture is passed through a membrane emulsification device into a continuous phase by adopting a membrane emulsification mode to form monodisperse liquid crystal droplets. The principle of the membrane emulsification device is shown in fig. 3, which mainly utilizes the dispersion technology based on membrane emulsification to realize the preparation of monodisperse liquid crystal droplets. The specific operation is as follows: the liquid crystal mixture as a disperse phase slowly passes through an inorganic membrane with micropores, liquid crystal droplets are formed after the liquid crystal mixture is extruded from the micropores of the inorganic membrane and are dispersed in a continuous phase, so that a dispersion system taking the liquid crystal droplets as the disperse phase is formed, the size of the liquid crystal droplets can be controlled by the pore size of the micropores of the inorganic membrane, and the particle size of the prepared polymer particles with porous structures is finally controlled. In the following examples of the present application, a membrane emulsification apparatus using an SPG membrane having micro pores is selected to precisely control the particle size of liquid crystal droplets. The continuous phase contains a liquid crystal conformation modifier which can make the liquid crystal molecules (including reactive liquid crystal and non-reactive liquid crystal) in the liquid crystal microdroplets regularly arranged. When the liquid crystal conformation change agent is an ionic surfactant (SDS) or a salt, liquid crystal molecules in the liquid crystal microdroplets are arranged along the radial direction of the liquid crystal microdroplets to form a ray-type conformation; when the liquid crystal conformation change agent is a nonionic surfactant (PVP, PVA, PEG, Pluronic, etc.), liquid crystal molecules in the liquid crystal droplets are arranged in a manner that the director is tangent to the circumference of the liquid crystal droplets to form a bipolar conformation.
The reactive liquid crystals in the droplets are then polymerized to form mesospheres comprising non-reactive liquid crystals that do not participate in the polymerization. Before polymerization, the liquid crystal molecules in the liquid crystal microdroplets are regularly arranged due to the presence of the liquid crystal conformation change agent, so that the internal structure of the formed intermediate microspheres substantially maintains the regular arrangement before reaction after polymerization. The polymerization may be photopolymerization, thermal polymerization or radiation polymerization. In the embodiment of the present invention, the polymerization method is preferably photopolymerization.
And finally, removing the non-reactive liquid crystal which does not participate in polymerization to further form the porous polymer particles with regular internal structures and pore channel arrangement. Since the non-reactive liquid crystal compound does not participate in the polymerization reaction, micropores are formed in the polymer particles after removal, and the distribution of the micropores tends to be regularly aligned due to the influence of the previous alignment of the liquid crystal molecules. These porous polymeric microparticles are substantially spherical in the swollen state, but when dried, the porous polymeric microparticles shrink to different shapes due to different internal structures and pore channel arrangements, for example, porous polymeric microparticles having a radial conformation substantially retain a spherical shape after shrinking, while porous polymeric microparticles having a bipolar conformation shrink to an ellipsoidal shape. Meanwhile, the distribution and the pore diameter of the inner pore canal of the prepared porous polymer microsphere can be influenced by the content of reactive liquid crystal and the concentration of a liquid crystal conformation modifier in a continuous phase.
And secondly, weighing a certain amount of prepared porous polymer particles, and dispersing the porous polymer particles in the homogenate for swelling to form suspension. The homogenate is an organic solvent commonly used in gel permeation chromatography, such as tetrahydrofuran, chloroform, toluene, or dimethylformamide, and the like. In the following examples, tetrahydrofuran was used for the homogenates.
And a third step of filling the suspension prepared in the second step into a hollow column tube to finally form a chromatographic column containing porous polymer particles. In an embodiment of the invention, the suspension is packed into the hollow column tubes using a homogenization process. In embodiments of the invention, the homogenization employed is the usual operating method and conditions for packing chromatographic columns.
The structure, preparation and separation effects of the column of the present invention and the method for preparing the same will be described in detail below with reference to specific examples. In the following examples, the percentages are by weight unless otherwise specified. In the detection of the chromatographic column, the instrument used is a Waters gel chromatograph, wherein the detector is a time-lapse refractometer. In the detection, the mobile phase used is tetrahydrofuran, and the column temperature is 35 ℃.
In the following examples, the general steps for forming a homogeneous liquid crystal mixture are: mixing the reactive liquid crystal, the non-reactive liquid crystal and the polymerization initiator according to a certain proportion, heating the mixture to a temperature above the clearing point of the mixed liquid crystal until the mixture becomes a uniform solution, fully vibrating the solution to mix the solution uniformly, and then slowly cooling the solution to room temperature to form a liquid crystal mixture. When photopolymerization is used, the solution must be kept in the dark while slowly cooling, since the photoinitiator is sensitive to light.
The general steps for forming the polymer microparticles are: at a certain speed, the uniform liquid crystal mixture slowly and smoothly passes through the SPG film emulsifying device to be dispersed into a continuous phase containing the conformation change agent, the stirring speed of the continuous phase is 300r/min, and finally, an emulsion containing liquid crystal microdroplets with uniform size is formed. The emulsion containing the liquid crystal microdroplets is placed under a 365nm UV light source for curing polymerization, and the radiation intensity is 2.5mW/cm2The polymerization time is 30 minutes, and the system needs to be stirred continuously in the polymerization process. After polymerization, the polymer particles were washed three times with an ethanol solution, and centrifuged to remove the supernatant, thereby obtaining polymer particles from which unreacted substances were removed.
Example 1
A liquid crystal mixture containing 9g of reactive liquid crystal RM257, 21g of non-reactive liquid crystal 5CB and 0.3g of photoinitiator DMPAP (the reactive liquid crystal accounts for 29.7 percent of the liquid crystal mixture) was prepared, and polymer particles were prepared according to the above steps, wherein the pore diameter of the SPG membrane micropore is 2.8 microns, the continuous phase is water, and the molar concentration of the liquid crystal conformation change agent SDS in the water is 78 mM. As shown in fig. 4, the average size of the polymer fine particles was 7 μm when dried (fig. 4 (a)). Meanwhile, since the liquid crystal conformation modifier is SDS, the polarization micrograph shows the optical anisotropy of the ray type (Maltese black cross) (FIG. 4 (b)).
The prepared porous polymer particles were then swollen in tetrahydrofuran and poured into a 7.8 x 300mm empty column tube by a conventional homogenization method to form a chromatography column. And finally, connecting the prepared chromatographic column to a testing instrument to test the separation effect. The log W-V calibration curve of the column was measured as shown in FIG. 5, indicating that the fractionation range of the prepared column was 500 to 48000.
Further, the mixed samples containing polystyrenes of different molecular weights were passed through the prepared column and a comparative column, respectively, under the same test conditions, wherein the comparative column was a commercially available gel column having a stationary phase, a separation range and a separation column size substantially the same as the particle size of the column prepared according to this example. As shown in fig. 6 and table 1, the retention time of the column of this example was reduced by 20% or more for each molecular weight compound in the mixed sample compared to the retention time of the comparative column.
TABLE 1
Example 2
A liquid crystal mixture containing 12g of reactive liquid crystal RM257, 18g of non-reactive liquid crystal 5CB and 0.3g of photoinitiator DMPAP (the reactive liquid crystal accounts for 39.6% of the liquid crystal mixture) was prepared, and polymer particles were prepared according to the above procedure, wherein the pore diameter of the SPG membrane micropores was 10 μm, the continuous phase was water, and the molar concentration of the liquid crystal conformation change agent SDS in water was 2 mM. The average size of the polymer particles was 25 microns on drying (FIG. 7(a)), while the swollen size in tetrahydrofuran averaged 30 microns (FIG. 7 (b)). Meanwhile, since the liquid crystal conformation modifier is SDS, the polarization micrograph shows optical anisotropy of the ray type (Maltese black cross). Meanwhile, as shown in fig. 7(c), the SEM image of the surface of the polymer fine particle further shows the porous characteristic thereof.
The prepared porous polymer particles were then swollen in tetrahydrofuran and poured into a 7.8 x 300mm empty column tube by a conventional homogenization method to form a chromatography column. And finally, connecting the prepared chromatographic column to a testing instrument for separation and detection. The log W-V calibration curve of the column was measured as shown in FIG. 8, and it was revealed that the prepared column was increased in the content of the reactive liquid crystal as compared with example 1 in the fractionation range of 500 to 48000, but the column was still able to achieve the fractionation range of 500 to 48000 by adjusting the concentration of SDS.
Example 3
A liquid crystal mixture containing 12g of reactive liquid crystal RM257, 18g of non-reactive liquid crystal 5CB and 0.3g of photoinitiator DMPAP (the reactive liquid crystal accounts for 39.6 percent of the liquid crystal mixture) was prepared, and polymer particles were prepared according to the above steps, wherein the pore diameter of the SPG membrane micropore is 2.8 microns, the continuous phase is water, and the molar concentration of the liquid crystal conformation change agent SDS in the water is 78 mM. The average size of the polymer microparticles was 7 μm when dried (see fig. 9 (a)). Meanwhile, since the liquid crystal conformation modifier is SDS, the polarization micrograph shows the optical anisotropy of the ray type (Maltese black cross) (see FIG. 9 (b)).
The prepared porous polymer particles were then swollen in tetrahydrofuran and poured into a 7.8 x 300mm empty column tube by a conventional homogenization method to form a chromatography column. And finally, connecting the prepared chromatographic column to a testing instrument to test the separation effect. The log W-V calibration curve of the column was measured as shown in FIG. 10, indicating that the fractionation range of the prepared column was 850 to 2400. The content of the reactive liquid crystal was increased as compared with example 1, so that the pore size of the polymer fine particles became small, and the classification range of the final column was reduced to 850-2400.
Example 4
A liquid crystal mixture containing 12g of reactive liquid crystal RM257, 18g of non-reactive liquid crystal 5CB and 0.3g of photoinitiator DMPAP (the reactive liquid crystal accounts for 39.6 percent of the liquid crystal mixture) was prepared, and polymer particles were prepared according to the above steps, wherein the pore diameter of the SPG membrane micropores was 20 microns, the continuous phase was water, and the molar concentration of a liquid crystal conformation change agent SDS in the water was 78 mM. The average size of the polymer microparticles when dried was 55 μm (see fig. 11 (a)). Meanwhile, since the liquid crystal conformation modifier is SDS, the polarization micrograph shows the optical anisotropy of the ray type (Maltese black cross) (see FIG. 11 (b)).
Example 5
A liquid crystal mixture containing 9g of reactive liquid crystal RM257, 21g of non-reactive liquid crystal 5CB and 0.3g of photoinitiator DMPAP (the reactive liquid crystal accounts for 39.6% of the liquid crystal mixture) was prepared, and polymer particles were prepared according to the above procedure, wherein the pore diameter of the SPG film micropores was 2.8 microns, the continuous phase was water, and the concentration of liquid crystal conformation change agent Pluronic in water was 1% by weight. As shown in FIGS. 12(a) and (b), the polymer particles were spheroids when dried, and the average size along the major axis of the spheroids was 10 μm. When swollen, the polymer particles swell in the direction of the minor axis of the ellipsoid, and finally form substantially spherical polymer particles.
The prepared porous polymer particles were then swollen in tetrahydrofuran and poured into a 4.6 x 250mm empty column tube using a conventional homogenization method to form a chromatography column. And finally, connecting the prepared chromatographic column to a testing instrument to test the separation effect. The logW-V calibration curve of the column is shown in FIG. 13, and the fractionation range of the column can reach 500-48000.
It can be seen from the data of the above example and the comparative column that the stationary phase formed by the polymer particles with regular internal structure and pore arrangement is introduced, and under the condition of ensuring the separation range, the separation time of the sample can be greatly shortened, thereby saving the mobile phase and improving the separation effect.
Although several exemplary embodiments have been described above in detail, the disclosed embodiments are merely exemplary and not limiting, and those skilled in the art will readily appreciate that many other modifications, adaptations, and/or alternatives are possible in the exemplary embodiments without materially departing from the novel teachings and advantages of the present disclosure. Accordingly, all such modifications, adaptations, and/or alternatives are intended to be included within the scope of the present disclosure as defined by the following claims.
Claims (10)
1. A chromatographic column for gel permeation chromatography, comprising a hollow column tube and a stationary phase distributed in the column tube, wherein the stationary phase is formed by stacking porous polymer particles, characterized in that the porous polymer particles have a regular internal structure and pore channel arrangement, and the porous polymer particles are substantially spherical when swollen.
2. The chromatographic column as claimed in claim 1, wherein the internal structure and pore channel arrangement of the porous polymeric microparticles have a radial conformation.
3. The chromatography column of claim 1, wherein the internal structure and pore arrangement of the porous polymeric microparticles have a bipolar conformation.
4. A chromatography column as claimed in any of claims 1 to 3, wherein the porous polymeric microparticles have an average particle size, when dry, in the range of from 5 microns to 100 microns.
5. A column according to any one of claims 1 to 3, wherein the porous polymeric microparticles are of an acrylate polymer.
6. A chromatography column according to any one of claims 1 to 3, having a fractionation range of 300 to 100000.
7. A method of preparing the chromatography column of claim 1, the method comprising:
(I) preparing porous polymer particles with a regular internal structure and pore channel arrangement;
(II) dispersing the porous polymer particles in a homogenate for swelling to form a suspension;
(III) packing the suspension into an empty column tube.
8. The method of claim 7, wherein the homogenate is selected from tetrahydrofuran, chloroform, toluene, or dimethylformyl.
9. The method of claim 7, wherein the step of preparing porous polymeric microparticles having a regular internal structure and pore channel arrangement further comprises:
1) forming a homogeneous liquid crystal mixture, wherein the liquid crystal mixture comprises reactive liquid crystals, non-reactive liquid crystals, and a polymerization initiator;
2) emulsifying the liquid crystal mixture to form liquid crystal droplets dispersed in a continuous phase containing a liquid crystal conformation change agent, wherein the liquid crystal conformation change agent can enable liquid crystal molecules in the liquid crystal droplets to be regularly arranged;
3) polymerizing the reactive liquid crystal in the liquid crystal microdroplets to form intermediate microspheres;
4) removing the non-reactive liquid crystal that is not polymerized from the intermediate microspheres to form the porous polymeric microparticles having a regular internal structure and pore channel arrangement.
10. The method of claim 9, wherein the reactive liquid crystal is present in an amount of 25% to 50% by weight of the total liquid crystal mixture.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811417535.3A CN111220745B (en) | 2018-11-26 | 2018-11-26 | Chromatographic column and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811417535.3A CN111220745B (en) | 2018-11-26 | 2018-11-26 | Chromatographic column and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111220745A true CN111220745A (en) | 2020-06-02 |
| CN111220745B CN111220745B (en) | 2023-05-09 |
Family
ID=70828798
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811417535.3A Active CN111220745B (en) | 2018-11-26 | 2018-11-26 | Chromatographic column and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111220745B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112684031A (en) * | 2020-12-04 | 2021-04-20 | 东莞西典医药科技有限公司 | HPLC (high Performance liquid chromatography) determination method for content of povidone K30 |
| CN114702732A (en) * | 2022-06-07 | 2022-07-05 | 江苏集萃智能液晶科技有限公司 | Polymer particle with double-size pore channel and preparation method thereof |
| CN114700055A (en) * | 2022-06-07 | 2022-07-05 | 江苏集萃智能液晶科技有限公司 | Chromatography medium and chromatography device |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3568840A (en) * | 1967-12-30 | 1971-03-09 | Mitsubishi Chem Ind | Packing materials for gel permeation chromatography |
| JPS5599068A (en) * | 1979-01-24 | 1980-07-28 | Sekisui Chem Co Ltd | Filler for liquid chromatography |
| CN1243761A (en) * | 1998-07-30 | 2000-02-09 | 武汉大学 | Process for preparing porous filler of regenerated cellulose for gel permeation chromatographic column |
| CN104014322A (en) * | 2014-05-14 | 2014-09-03 | 四川师范大学 | Dispersive polymeric microsphere fixed phase padding with narrow grain diameter and preparation method thereof |
| CN104028252A (en) * | 2014-05-20 | 2014-09-10 | 四川师范大学 | Stationary-phase filler with polyacrylate matrix and preparation method thereof |
| CN109954454A (en) * | 2017-12-25 | 2019-07-02 | 江苏集萃智能液晶科技有限公司 | With optically anisotropic porous structure polymer microballoon and its preparation method and application |
-
2018
- 2018-11-26 CN CN201811417535.3A patent/CN111220745B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3568840A (en) * | 1967-12-30 | 1971-03-09 | Mitsubishi Chem Ind | Packing materials for gel permeation chromatography |
| JPS5599068A (en) * | 1979-01-24 | 1980-07-28 | Sekisui Chem Co Ltd | Filler for liquid chromatography |
| CN1243761A (en) * | 1998-07-30 | 2000-02-09 | 武汉大学 | Process for preparing porous filler of regenerated cellulose for gel permeation chromatographic column |
| CN104014322A (en) * | 2014-05-14 | 2014-09-03 | 四川师范大学 | Dispersive polymeric microsphere fixed phase padding with narrow grain diameter and preparation method thereof |
| CN104028252A (en) * | 2014-05-20 | 2014-09-10 | 四川师范大学 | Stationary-phase filler with polyacrylate matrix and preparation method thereof |
| CN109954454A (en) * | 2017-12-25 | 2019-07-02 | 江苏集萃智能液晶科技有限公司 | With optically anisotropic porous structure polymer microballoon and its preparation method and application |
Non-Patent Citations (2)
| Title |
|---|
| XIAOGUANG WANG等: "Synthesis of Optically Complex, Porous, and Anisometric Polymeric Microparticles by Templating from Liquid Crystalline Droplets", 《ADVANCED FUNCTIONAL MATERIALS》 * |
| 刘英超等: "《中枢神经系统疾病蛋白质组学》", 30 September 2010, 山东科学技术出版社 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112684031A (en) * | 2020-12-04 | 2021-04-20 | 东莞西典医药科技有限公司 | HPLC (high Performance liquid chromatography) determination method for content of povidone K30 |
| CN112684031B (en) * | 2020-12-04 | 2022-05-17 | 东莞西典医药科技有限公司 | HPLC (high Performance liquid chromatography) determination method for content of povidone K30 |
| CN114702732A (en) * | 2022-06-07 | 2022-07-05 | 江苏集萃智能液晶科技有限公司 | Polymer particle with double-size pore channel and preparation method thereof |
| CN114700055A (en) * | 2022-06-07 | 2022-07-05 | 江苏集萃智能液晶科技有限公司 | Chromatography medium and chromatography device |
| CN114702732B (en) * | 2022-06-07 | 2022-08-26 | 江苏集萃智能液晶科技有限公司 | Polymer particle with double-size pore channel and preparation method thereof |
| WO2023236488A1 (en) * | 2022-06-07 | 2023-12-14 | 江苏集萃智能液晶科技有限公司 | Chromatography medium and chromatography device |
| WO2023236487A1 (en) * | 2022-06-07 | 2023-12-14 | 江苏集萃智能液晶科技有限公司 | Polymeric microparticle having pore channels of two sizes and preparation method therefor |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111220745B (en) | 2023-05-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111220745B (en) | Chromatographic column and preparation method thereof | |
| Svec et al. | Design of the monolithic polymers used in capillary electrochromatography columns | |
| Zhou et al. | Synthesis of macroporous poly (glycidyl methacrylate) microspheres by surfactant reverse micelles swelling method | |
| Wang et al. | Synthesis of optically complex, porous, and anisometric polymeric microparticles by templating from liquid crystalline droplets | |
| CN109423300B (en) | Process for producing monodisperse polymer microparticles | |
| JP6733900B2 (en) | Separation carrier, method for producing separation carrier, column, and device for liquid chromatography or solid phase extraction | |
| EP0862737A1 (en) | Chromotographic method and device in which a continuous macroporous organic matrix is used | |
| CN111434610B (en) | Porous silicon dioxide microsphere and preparation method and application thereof | |
| CN110964142B (en) | Chiral polymer microsphere with porous structure and preparation method thereof | |
| Guo et al. | Preparation of monodispersed porous polyacrylamide microspheres via phase separation in microchannels | |
| EP2027163B1 (en) | A method for producing cross-linked spherical particles | |
| Ousalem et al. | Evaluation of the porous structures of new polymer packing materials by inverse size-exclusion chromatography | |
| Huš et al. | Tailoring morphological features of cross-linked emulsion-templated poly (glycidyl methacrylate) | |
| Chen et al. | Graft copolymers having hydrophobic backbone and hydrophilic branches. xvi. Polystyrene microspheres with poly (N‐isopropylacrylamide) branches on their surfaces: size control factors and thermosensitive behavior | |
| CN112246197B (en) | Porous structure polymer microsphere with optical anisotropy and application thereof | |
| CN110117341B (en) | Preparation method of porous functional polymer microspheres | |
| Gölgelioğlu et al. | Aqueous size exclusion chromatography in semimicro and micro-columns by newly synthesized monodisperse macroporous hydrophilic beads as a stationary phase | |
| JPH11189601A (en) | Manufacture of fine polymer particle and manufacture of filler for liquid chromatography | |
| Tikhonov et al. | New approach to the synthesis of a functional macroporous poly (vinyl alcohol) network and design of boronate affinity sorbent for protein separation | |
| Gwon et al. | General method for the production of hydrogel droplets from uniformly sized smart shell membranes | |
| Zhang | Progress in preparation of monodisperse polymer microspheres | |
| CN110553981B (en) | Method for detecting content of organic solvent in mixed solution | |
| TWI824976B (en) | Method for producing stationary phase medium of porous microspheres | |
| Kibar et al. | Synthesis and characterization of monodisperse-porous, zwitterionic microbeads | |
| Jin et al. | Monosized macroporous resins with epoxy groups and their structure and gel chromatography properties |
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 |