CN111729661A - Chromatography media for separating boron-containing species - Google Patents
Chromatography media for separating boron-containing species Download PDFInfo
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- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
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- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 - B01D15/36
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Abstract
The invention relates to the field of chromatography media, in particular to a chromatography medium for separating boron-containing substances. The chromatography medium comprises a base support and grafted chains grafted to the base support; the general formula of the grafting chain is MA-R1‑X1‑R2Or
Shown; wherein MA is a group that can be linked to other chemical functional groups, either covalently or non-covalently; r1、R2And R3Independently selected from hydrogen atoms, saturated hydrocarbon groups, unsaturated hydrocarbon groups or saturated hydrocarbon groups containing heteroatoms which are not carbon electronegative, and at least one of the groups carries cis-ortho-polyhydroxy, wherein the cis-ortho-polyhydroxy is a linear chain skeleton structure or a ring chain skeleton structure;X1and X2Divalent and trivalent heteroatoms of said non-carbon electronegative heteroatom, respectively; r in the graft chain1、R2And R3Sum of number of carbon atoms present therein>2 pieces of the Chinese herbal medicines.
Description
Technical Field
The invention relates to the field of chromatography media, in particular to a chromatography medium for separating boron-containing substances.
Background
Borax is commonly used as a medicine in ancient times of China. There are many descriptions of the properties of Chinese medicine: borax, also known as Yueshi, is sweet, salty and cool in nature, and enters lung and stomach meridians. Has effects of clearing away heat and toxic materials, relieving swelling, preventing putrefaction, clearing lung-heat, and eliminating phlegm, and can be used for treating sore throat, aphtha, conjunctival congestion, nebula, phlegm heat, and cough. Boron atoms have unique properties that are different from the atoms that are often present in other drug molecules. The boron atom center may be from sp2Hybrid triangular plane becomes sp3The hybrid tetrahedron and boron-containing compound increase the diversity of the molecular structure of the medicine. The compound becomes a new way for drug development by virtue of a brand-new structure and a special action mechanism. With the development of boron chemistry, more and more organoboron compounds having better biological activity have been successively synthesized in recent years. They have good effects in resisting tumor, resisting infection, inhibiting thrombin and dipeptidyl peptidase activities, etc.
At present, many kinds of boron-containing medicines are industrialized, and the commercial value is high. Including Tavaborole (trade name Kerydin), criborole (trade name eucristole), VABOMERE complex, boracic derivatives, boracic agents and oligonucleotides, Bortezomib (trade name Velcade), ixazomide (trade name Ninlaro).
Among them, tavaborole and criborole are new molecular entities from Anacor that were approved by the U.S. FDA for topical use in 2014 and 2016, respectively, for the treatment of onychomycosis (onychomycosis/fingernail) and atopic dermatitis (eczema), respectively. VABOMERE consists of Meropenem (Meropenem, an antibacterial drug) and vaboractam (which can inhibit resistance to certain bacteria), approved by the U.S. FDA for treatment of adult complicated urinary tract infections in 2017. Vaborbacam is a cyclic boronic acid pharmacophore beta-lactamase inhibitor and Vabomere can cope with infections by beta-lactamase producing gram negative bacteria. All three drugs are pioneered organoborates. And bortezomib and ethacryl are also organic boron drugs containing dipeptidyl groups. Bortezomib, marketed in 2003 for the treatment of multiple myeloma, is a proteasome inhibitor; efalazomib was marketed in 2015 as the first oral drug for the treatment of multiple myeloma, a second generation proteasome inhibitor.
As a medicine, the most critical point of quality control is the control of medicine purity, so that the separation and purification of the medicine in production are critical processes, and the separation efficiency and the process not only influence the medicine quality but also influence the medicine production cost. At present, the production and purification methods of the boron-containing drug molecules are generally crystallization purification and reverse phase chromatography purification. The advantage of such purification methods is the simplicity of operation, but several serious disadvantages: a large amount of organic solvents are often needed in the purification process, and potential safety hazards such as fire, explosion and the like may exist in the use process of the organic solvents; the organic solvent is easy to remain in the medicine and has certain harm to human body; the subsequent treatment of organic solvent and reverse phase chromatography silica gel medium can cause great damage to the environment; the method has low biocompatibility and is easy to cause the denaturation and inactivation of biological medicines.
Disclosure of Invention
The invention aims to fill the blank of the prior purification technology and provide an affinity chromatography medium for separating or enriching boron-containing micromolecules, oligonucleotides, polypeptides and proteins;
the invention relates to a chromatographic medium for separating boron-containing substances, which comprises a matrix support and a grafting chain grafted on the matrix support; the invention also aims to provide a preparation method of the affinity chromatography medium; the affinity chromatography medium is used for the analysis application of a chromatographic column; the application of the affinity chromatography medium in industrial separation and purification. The prepared chromatographic medium has high separation efficiency, good separation effect and low comprehensive total cost, and is suitable for industrialized large-scale separation and purification of boron-containing micromolecules, oligonucleotides, polypeptides, proteins and the like.
Specifically, the chromatographic medium for separating boron-containing substances provided by the invention comprises a matrix support and a grafting chain grafted on the matrix support;
the general formula of the grafting chain is MA-R1-X1-R2Or
Shown;
wherein MA is a group that can be linked to other chemical functional groups, either covalently or non-covalently;
R1、R2and R3Independently selected from hydrogen atoms, saturated hydrocarbon groups, unsaturated hydrocarbon groups or saturated hydrocarbon groups containing heteroatoms which are not carbon electronegative, and at least one of the groups carries cis-ortho-polyhydroxy, wherein the cis-ortho-polyhydroxy is a linear chain skeleton structure or a ring chain skeleton structure;
X1and X2Divalent and trivalent heteroatoms of said non-carbon electronegative heteroatom, respectively;
r in the graft chain1、R2And R3Sum of number of carbon atoms present therein>2 pieces of the Chinese herbal medicines.
The invention also relates to a chromatographic separation device containing a chromatographic medium as described above.
According to a further aspect of the invention, the invention also relates to the use of a chromatography medium as described above, or a chromatography column as described above, for separating or enriching a boron-containing substance from a liquid medium.
The invention has the beneficial effects that:
1) has specific binding effect on boron-containing molecules;
2) the conditions of separation and purification are mild/simple, the biocompatibility is high, and the biological activity of purified molecules can be well maintained;
3) the purified chromatographic medium and the chromatographic column can be repeatedly used after being cleaned;
4) does not need to use a large amount of organic reagent;
5) less waste liquid is generated, the treatment is easy, and the environmental pollution is reduced.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a scanning electron micrograph of a chromatographic medium according to one embodiment of the present invention;
FIG. 2 is a graph of the particle size distribution of a chromatographic medium in one embodiment of the invention;
FIG. 3 is a chromatogram of the separation of boron-containing molecules from non-boron-containing molecules by a chromatographic medium in accordance with an embodiment of the present invention;
FIG. 4 is a chromatogram of the adsorption effect of a chromatographic medium on a boron-containing molecule in a comparative example of the present invention;
FIG. 5 is a chromatogram of the separation and purification of a bortezomib sample with a chromatographic medium in accordance with an embodiment of the present invention;
FIG. 6 is a chromatogram of the separation of a bortezomib sample with a chromatographic medium in a control example of the invention.
Detailed Description
Reference will now be made in detail to embodiments of the invention, one or more examples of which are described below. Each example is provided by way of explanation, not limitation, of the invention. In fact, it will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the scope or spirit of the invention. For instance, features illustrated or described as part of one embodiment, can be used on another embodiment to yield a still further embodiment.
It is therefore intended that the present invention cover the modifications and variations of this invention provided they come within the scope of the appended claims and their equivalents. Other objects, features and aspects of the present invention are disclosed in or are apparent from the following detailed description. It is to be understood by one of ordinary skill in the art that the present discussion is a description of exemplary embodiments only, and is not intended as limiting the broader aspects of the present invention.
The present invention relates to a chromatography medium for separating boron-containing species, said chromatography medium comprising a base support and polymer chains grafted to said base support;
the polymer chain has the general formula MA-R1-X1-R2Or
Shown;
wherein MA is a group that can be linked to other chemical functional groups, either covalently or non-covalently;
R1、R2and R3Independently selected from hydrogen atoms, saturated hydrocarbon groups, unsaturated hydrocarbon groups or saturated hydrocarbon groups containing heteroatoms which are not carbon electronegative, and at least one of the groups carries cis-ortho-polyhydroxy, wherein the cis-ortho-polyhydroxy is a linear chain skeleton structure or a ring chain skeleton structure;
X1and X2Divalent and trivalent heteroatoms of said non-carbon electronegative heteroatom, respectively;
r in the graft chain1、R2And R3Sum of number of carbon atoms present therein>2 pieces of the Chinese herbal medicines.
The separation principle of the invention is based on the fact that boric acid and cis-diol are combined through a fast and stable bond to form a borate, the covalent B-O bond of the borate is stable, but the formation of the B-O bond is reversible under certain conditions or under the action of certain external stimuli. The principle has wider application in boric acid affinity chromatography, namely, phenylboronic acid is connected to a matrix framework to carry out affinity adsorption on molecules with cis-diol bonds. The present invention innovatively develops a reverse application based on the above principle, i.e., bonding groups with cis-ortho-polyhydroxy groups to the matrix backbone for affinity adsorption of boron-containing molecules. The invention skillfully passes the group with cis-ortho-polyhydroxy through a non-carbon with charge negativityX(X1Or X2) The heteroatom is bonded to the matrix backbone; the X heteroatom is connected to a spacer arm with a proper length to increase the flexibility of the ortho-polyhydroxy group (active group), so that the ortho-polyhydroxy group (active group) is easier to be combined with the boron atom in space to form effective coordination; on one hand, lone pair electrons on the X heteroatom can form intramolecular hydrogen bonds with hydroxyl in the active group, and the ortho-position polyhydroxy group forms cis-form conformation which is favorable for combining boron atoms; the lone pair of electrons on the X heteroatom can interact with the boron atom on the other hand, and the molecule containing the boron atom is drawn to the periphery of the group with cis-ortho polyhydroxy; the synergy of the multiple actions makes the ortho-polyhydroxy more easily combined with boron atoms in space and conformation to form effective coordination.
In some embodiments, the non-carbon electronegative heteroatom has a strong electronegativity.
In some embodiments, the non-carbon electronegative heteroatoms each have a pauling electronegativity value of greater than 2.0, preferably greater than 2.1, more preferably greater than 2.5, and more preferably greater than 2.9.
In some embodiments, the non-carbon electronegative heteroatom is selected from one or more of O, S, N, P.
In some embodiments, the saturated hydrocarbyl group containing a non-carbon electronegative heteroatom may be a polyamine, an oligoimine, a polyether, an oligoether, a polythioether, an oligosulfide; more preferred are polyamines and oligoimines, such as: ethylenediamine, diethylenetriamine, dipropylenetriamine, triethylenetetramine, linear and branched oligoethyleneimines.
In some embodiments, X1Selected from O or S atoms.
In some embodiments, X2Selected from N or P atoms.
The number of cis-ortho polyols is greater than 2, for example 3,4, 5, 6, 7, 8, 9, 10 or more.
In some embodiments, R in the grafted chain1、R2And R3The sum of the number of carbon atoms in the above is 3 to 40, for example, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, and 13, 14, 15, 20, 25, 30 and 35 can be selected.
The saturated or unsaturated hydrocarbon group may be Cx-CyAn alkyl group. The term "Cx-CyAlkyl "refers to an alkyl group having from x to y (inclusive) carbon atoms in a branched or unbranched hydrocarbon group. By way of illustration, and not limitation, the term "C1-C4Alkyl "refers to a straight or branched chain hydrocarbon moiety having from 1 to 4 carbon atoms and includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. The term "C1-C4N-alkyl "refers to a straight chain hydrocarbon moiety having from 1 to 4 carbon atoms, including methyl, ethyl, n-propyl, and n-butyl. Cx-CyWhere x may be from 1 to 10 and y is from 2 to 20. Each alkyl, cycloalkyl group may be optionally substituted such as, but not limited to, as indicated herein. In some embodiments, the group is mono-or di-substituted. In some embodiments, alkyl is C1-C3、C1-C4、C1-C6、C4-C6Or C1-C10An alkyl group.
In some embodiments, the ring chain backbone structure may also be a cycloalkyl group. Such as, but not limited to, cyclopropylmethyl, dimethylcyclopropyl, cyclopropylethyl, cyclopropylpropyl, cyclopropylbutyl, cyclobutylmethyl, cyclobutylethyl, cyclobutylpropyl, cyclopentylmethyl, cyclopentylethyl, cyclopentylpropyl, cyclohexylmethyl, cyclohexylethyl, and cyclohexylpropyl.
Each alkyl, cycloalkyl group may be optionally substituted or modifiable, such as but not limited to as indicated herein. In some embodiments, the group is mono-or di-substituted.
In some embodiments, the cyclic backbone structure is a monosaccharide, disaccharide, or polysaccharide;
in some embodiments, the cyclic chain backbone structure is preferably mannose, fructose, galactose, mannose-containing disaccharides, and mannose-containing oligosaccharides.
The matrix support comprises sites MA which can be chemically modified. May be covalently or non-covalently linked to other chemical functional groups. These modification sites include, but are not limited to, the following chemical functional groups: MA is selected from at least one of epoxy group, amino group, aldehyde, hydroxyl group, carboxyl group, oxo group and thiol group. The modification sites typically also comprise derivatising groups with these corresponding chemical functionalities, for example: amino derivatives, thiol derivatives, aldehyde derivatives, formyl derivatives, biotin derivatives, alkyne derivatives, hydroxyl derivatives, activated hydroxyl or derivatives, carboxylate derivatives, activated carbonates, activated esters, NHS esters (succinimidyl), NHS carbonates (succinimidyl), imido esters or derivatives, cyanobromide derivatives, maleimide derivatives, haloacetyl derivatives, iodoacetamide/iodoacetyl derivatives, epoxy derivatives, streptavidin derivatives, Tresyl derivatives, diene/conjugated diene derivatives (Diels-Alder type reactions), alkene derivatives, substituted phosphate derivatives, bromohydrin/halohydrin, substituted disulfides, pyridyl-disulfide derivatives, azalides, hydride derivatives, halobenzene derivatives, nucleoside derivatives, branched/multifunctional linker molecules, dendrimers and derivatives, nucleoside derivatives, or any combination thereof.
In some embodiments, the density of the surface chemical modification sites on the chromatographic medium is optimizable. For example, >0.1mmol/ml medium, preferably >0.5mmol/ml medium, more preferably >1.0mmol/ml medium, more preferably >2.0mmol/ml medium.
In some embodiments, R1、R2And R3At least one of which carries a cis-ortho-polyol; preferably a cis-vicinal diol; more preferably a terminal cis-vicinal diol;
in some embodiments, R1、R2And R3At least one of which carries a catechol, such as a catechol homolog.
In some embodiments, the matrix support comprises any one of borosilicate glass, agar, agarose derivatives, magnetic beads, silica, titanium dioxide, alginate, cellulose derivatives, dextran, starch, cyclodextrin, chitosan, carrageenan, guar gum, gum arabic, gum ghatti, gum tragacanth, karaya gum, locust bean gum, xanthan gum, pectin, mucin, liver thioesters and gelatin, silicon, ceramics, glass, polyurethane, polystyrene divinyl benzene, polymethyl methacrylate, polyacrylamide, polyethylene glycol terephthalate, polyvinyl acetate, polyethylene, polypropylene, polyvinyl chloride, polyvinyl pyrrolidone, or copolymers formed of any of several.
The substrate support may be selected from a range of options of suitable shapes, forms, materials and variations depending on the application. The substrate support surface may be substantially flat or planar. Or may be circular or the like. The shape of the surface of the substrate support includes, but is not limited to, a hole, a depression, a pillar, a ridge, a channel, a membrane, or the like. The substrate support is preferably a particulate material having a substantially planar surface. The shape of the particulate matter is preferably spherical or substantially spherical.
In some embodiments, the matrix support is in the form of particles having a particle size in the range of 1 μm to 500 μm.
In some embodiments, the particle size ranges from 1 μm to 300 μm;
in some embodiments, the particle size ranges from 2 μm to 200 μm.
Further, the matrix support may be non-porous or include one or more pores. In particular, a porous resin may be used, and examples of the porous resin may include: other agarose-based resins (e.g., Sepharose Fast Flow series (GE)), polymethacrylates: (Sepax PGMA series), polystyrene divinylbenzene (Sepax PolyRP and Sepax Proteomix series), Silica (Sepax Silica series), controlled pore glass, dextran derivatives, polyacrylamide derivatives, and/or other polymers; or any combination thereof. The non-porous resin may be prepared from the same matrix material as the above porous resin.
In some embodiments, the pore size is
In some embodiments, the pore size is
In some embodiments, the pore size is
In the context of the present invention, "non-porous" means that the substrate support has no appreciable pores, e.g.pore diameter
Further, the products are exemplified by the different types of chromatographic media products of Seikagaku corporation, including but not limited to the listed types:
(a) chromatography media having covalently attached epoxy groups: polar MC-Epoxy (porous polydisperse polymethacrylate series), Monomix MC-Epoxy (porous monodisperse polymethacrylate series), Proteomix Epoxy (nonporous monodisperse cross-linked polystyrene series), Proteomix POR-Epoxy (porous monodisperse cross-linked polystyrene series).
(b) Chromatography media having covalently attached other groups: including but not limited to-OH (hydroxyl), -CHO (aldehyde group), IEX (anion, cation exchange group), HIC (Hydrophobic group), IMAC (metal chelating group), Affinity, Hydrophilic-Hydrophobic balanced (Hydrophilic Hydrophobic amphoteric group), Mixed-mode (Mixed acting group).
The present invention can develop various chromatographic separation devices using a chromatographic medium containing the above-mentioned components. There are many types of chromatographic separation devices, including but not limited to the following: SPE solid phase extraction column, centrifuge tube with separating membrane, magnetic bead, separating membrane (membranes), fast detection biochip (bio-chips), kit, fiber bundle column, monolithic column and conventional analytical grade or preparative grade chromatographic column.
The chromatographic separation device is preferably cylindrical and consists of a number of essential components: chromatography media material, particle size and pore size of the chromatography media, matrix material of the column, size of the column, and method of packing the column. The chromatographic column filled by the chromatographic medium can have various physical forms and is formed by combining different essential components of the chromatographic column.
The method for assembling the column comprises the following steps: constant current/variable current/constant voltage/multi-stage pressure regulating/DAC.
Different column packing methods are selected according to the performances of the medium particle size, the aperture and the like and the specification of the chromatographic column.
The chromatographic medium with the grain diameter less than 15 mu m is packed into a column by a multi-purpose constant-current and constant-pressure method;
the chromatographic medium with the particle size larger than 15 μm is packed into a column by a constant-current or variable-current method according to the pressure resistance of a column bed and the chromatographic medium;
the large-size chromatographic column is assembled by a multi-level pressure regulation or DAC method.
Homogenizing: water, brine, water or buffer salts containing the organic phase, etc., 20-80% (v: v) solids content
Selecting proper homogenate mobile phase and homogenate volume according to chromatographic column specification and chromatographic medium performance
Chromatography column material: stainless steel, glass, plastic and SPE tube
The column tubes made of different materials have different pressure resistance, and the maximum pressure resistance of the material of the column tube is not exceeded while the pressure is controlled during the filling of the chromatographic column.
Chromatographic column cleaning conditions: the long service life of the column adsorbs difficult to clean impurities which affect the performance of the column and require periodic cleaning. Different impurity cleaning methods are different, the common impurity is 0.5MHCl or 0.5-1.0M NaOH, and the impurity with strong hydrophobic combination can be cleaned by 0.1-1% Tween or Triton X-100. Alternatively, the column may be washed with 0.5M NaOH and stored in 20% aqueous ethanol at room temperature.
The chromatographic column preservation method comprises the following steps:
the column was kept in 20% ethanol in water at room temperature.
The chromatography medium is stored in 20% ethanol water solution at 2-8 deg.C.
According to a further aspect of the invention, the invention also relates to the use of a chromatographic medium as described above, or a chromatographic separation device as described above, for separating or enriching a boron-containing substance from a liquid medium.
The removing of the boron-containing species comprises: separation and purification between boron-containing molecules and boron-free molecules (affinity chromatography) and separation and purification between different boron-containing molecules (not limited to the method of using the affinity chromatography medium).
In some embodiments, the chromatographic medium adsorbs the boron-containing species under basic conditions and elutes the boron-containing species under acidic conditions.
The pH of the alkaline conditions may be >7.0, preferably pH >7.5, more preferably pH > 8.0.
The pH of acidic conditions may be <7.0, preferably pH <6.0, more preferably pH < 5.0.
In addition, in the presence of Mg2+ ions, the interaction between the boron-containing species and the cis-diol group is enhanced. A carbohydrate-containing mobile phase (e.g., sorbitol) may also be used to elute the boron-containing species.
In some embodiments, the boron-containing species comprises the following:
a) boron-containing compounds for proteasome inhibitors, such as bortezomib, efavir;
b) boron-containing, anti-fungal, bacterial, viral, organoboronic drug molecules such as tavaborole, kreb's and vaborbacam-Meropenem;
c) boron compound carriers, such as Boron-containing acridine derivatives, for use in Neutron Capture tumor Therapy (BNCT);
d) a boron-containing compound having thrombin inhibitory activity;
e) boron-containing oligonucleotides, polypeptides, proteins having therapeutic effects;
f) boron compounds for use in boron selective reactions for molecular modular synthesis.
g) For the recovery of unreacted or excess boron-containing reagent in a chemical reaction, preferably for solid surface or polymer and soluble boron-containing reagent reactions.
Boron compounds for use in boron selective reactions for molecular modular synthesis are well known to those skilled in the art, for example "Liang Xu, et al, chem.soc.rev.,2015, 44 (24): 8848-8858 "and" Raphael Oeschager, et al, Science,2020,368, 736-741 ". Boron-containing intermediates for organic synthesis such as organic halides containing masked borane groups and di-or polyboron compounds; boron selective reaction refers to chemical conversion in which two or more borane groups are present in the reactants and their reactivity can be distinguished; thus, more reactive borane groups can be selectively converted while the relatively inert borane groups remain intact. Inert boryl groups can generally be used under different reaction conditions and converted to other functional groups, thus providing a continuous and flexible synthesis for complex molecules.
Embodiments of the present invention will be described in detail with reference to examples.
Example 1
Amine diol example, epoxy grafting from media surface
In this example, porous polymethacrylate microspheres (particle size 30 μm, pore size 30 μm) produced by Suzhou Seiko technology Ltd were used
The goods number is: 281030952) having a large number of epoxy groups on the surface.
1) Adding 60.0g of 65-70% ethylamine aqueous solution by weight percentage based on 100g of chromatographic medium with epoxy group on the surface, adding into a reaction device, and mechanically stirring and reacting for 16h at room temperature;
2) after the reaction is finished, the washing medium is pumped and filtered to be neutral by deionized water; the chromatography medium was then transferred to 300ml of 0.05M hydrochloric acid solution and stirred well. Dipping a small amount of liquid by using pH test paper to detect that the pH is less than 3, and if the pH is not less than 3, continuously adding a hydrochloric acid solution, stirring until the pH is less than 3, filtering and draining; filtering the washing medium with deionized water to neutrality;
3) the chromatography medium is transferred to 500ml of 0.1M aqueous sodium bicarbonate solution and stirred well. Dipping a small amount of liquid by using pH test paper to detect that the pH is more than 8, and if the pH is not more than 8, continuously adding a sodium bicarbonate aqueous solution, stirring until the pH is more than 8, filtering and draining; washing with purified water for several times until the pH test paper is neutral, and draining until no filtrate comes out.
4) Adding 150ml of solution containing 50% of glycidol and 2.0g of sodium hydroxide into the medium, adding the mixture into a reaction device, mechanically stirring the mixture for reaction for 2 hours at room temperature under the condition of keeping out of the sun, and leaching and washing the medium by using deionized water with at least 10 times of volume after the reaction is finished.
The chromatography medium prepared in this example contains cis-diol structure that has an adsorption effect on boron-containing molecules under alkaline conditions, and the sample molecules are eluted under acidic conditions.
The scanning electron microscope of the examples is shown in FIG. 1, and the particle size distribution is shown in FIG. 2.
Example 2
In this example, porous polymethacrylate microspheres (particle size 30 μm, pore size 30 μm) produced by Suzhou Seiko technology Ltd were used
The goods number is: 281030952) having a large number of epoxy groups on the surface.
Adding 276g of methylamine water solution with the mass fraction of 30 percent into 100g of chromatographic medium with epoxy groups on the surface, adding into a reaction device, and mechanically stirring and reacting for 16h at room temperature; other procedures refer to example 1.
Example 3
In this example, porous polymethacrylate microspheres (particle size 30 μm, pore size 30 μm) produced by Suzhou Seiko technology Ltd were used
The goods number is: 281030502) having a large number of epoxy groups on the surface.
Adding 276g of methylamine water solution with the mass fraction of 30 percent into 100g of chromatographic medium with epoxy groups on the surface, adding into a reaction device, and mechanically stirring and reacting for 16h at room temperature; other procedures refer to example 1.
Example 4
In this example, porous polymethacrylate microspheres (particle size 30 μm, pore size 30 μm) produced by Suzhou Seiko technology Ltd were used
The goods number is: 281030502) having a large number of epoxy groups on the surface.
Adding 276g of methylamine water solution with the mass fraction of 30 percent into 100g of chromatographic medium with epoxy groups on the surface, adding into a reaction device, and mechanically stirring and reacting for 16h at room temperature; other procedures refer to example 1.
Example 5
In this example, porous polymethacrylate microspheres (particle size 60 μm, pore size 60 μm) produced by Suzhou Seiko technology was used
The goods number is: 281060952) having a large number of epoxy groups on the surface.
Adding 60.0g of 65-70% ethylamine aqueous solution by weight percentage based on 100g of chromatographic medium with epoxy group on the surface, adding into a reaction device, and mechanically stirring and reacting for 16h at room temperature; other procedures refer to example 1.
Example 6
In this example, porous polymethacrylate microspheres (particle size 60 μm, pore size 60 μm) produced by Suzhou Seiko technology was used
The goods number is: 281060952) having a large number of epoxy groups on the surface.
Adding 276g of methylamine water solution with the mass fraction of 30 percent into 100g of chromatographic medium with epoxy groups on the surface, adding into a reaction device, and mechanically stirring and reacting for 16h at room temperature; other procedures refer to example 1.
Example 7
In this example, porous polymethacrylate microspheres manufactured by Suzhou Seiko technologies, Inc. were used(particle diameter of 10 μm, pore diameter
The goods number is: 281010952) having a large number of epoxy groups on the surface.
Adding 60.0g of 65-70% ethylamine aqueous solution by weight percentage based on 100g of chromatographic medium with epoxy group on the surface, and mechanically stirring for reaction at room temperature for 16 h; other procedures refer to example 1.
Example 8
In this example, porous polymethacrylate microspheres (particle size 10 μm, pore size 10 μm) produced by Suzhou Seiko technology, Inc. were used
The goods number is: 281010952) having a large number of epoxy groups on the surface.
Adding 165.6g of methylamine water solution with the mass fraction of 30 percent into 100g of chromatographic medium with epoxy groups on the surface, adding the mixture into a reaction device, and mechanically stirring the mixture at room temperature for reaction for 16 hours; other procedures refer to example 1.
Example 9 (comparative example)
This example is a comparative example, where the surface of the chromatography medium has common cis-diol groups, but is not further modified. Porous polymethacrylate microspheres (particle size 30 μm, pore size 30 μm) produced by Suzhou Sai Kezhi Co Ltd
The goods number is: 281030952) having a large number of epoxy groups on the surface thereof, the epoxy groups being opened to form cis-diol groups.
Adding 300ml of 0.5M sulfuric acid into 100g of chromatographic medium with epoxy groups on the surface, adding the mixture into a reaction device, and mechanically stirring the mixture for reaction for 2 hours at the temperature of 40 ℃; after the reaction is finished, the washing medium is filtered by deionized water until the washing medium is neutral.
Example 10
In this example, porous polystyrene divinylbenzene (particle diameter: 15 μm, pore diameter: Suzhou Seiki technology, Ltd.) was used
The goods number is: 221715952) having a large number of epoxy groups on the surface.
Adding 60.0g of 65-70% ethylamine aqueous solution by weight percentage based on 100g of chromatographic medium with epoxy group on the surface, adding into a reaction device, and mechanically stirring and reacting for 16h at room temperature; other procedures refer to example 1.
Example 11
In this example, porous polystyrene divinylbenzene (particle diameter: 15 μm, pore diameter: Suzhou Seiki technology, Ltd.) was used
The goods number is: 221715952) having a large number of epoxy groups on the surface.
Adding 165.6g of methylamine water solution with the mass fraction of 30 percent into 100g of chromatographic medium with epoxy groups on the surface, adding the mixture into a reaction device, and mechanically stirring the mixture at room temperature for reaction for 16 hours; other procedures refer to example 1.
Example 12
This example used non-porous polystyrene divinylbenzene (particle size: 5 μm, non-porous, cat # 221605002) produced by Suzhou Seiki technology, Inc., which contained a large amount of epoxy groups on the surface.
Adding 60.0g of 65-70% ethylamine aqueous solution by weight percentage based on 100g of chromatographic medium with epoxy group on the surface, adding into a reaction device, and mechanically stirring and reacting for 16h at room temperature; other procedures refer to example 1.
Example 13
This example used non-porous polystyrene divinylbenzene (particle size: 5 μm, non-porous, cat # 221605002) produced by Suzhou Seiki technology, Inc., which contained a large amount of epoxy groups on the surface.
Adding 165.6g of methylamine water solution with the mass fraction of 30 percent into 100g of chromatographic medium with epoxy groups on the surface, adding the mixture into a reaction device, and mechanically stirring the mixture at room temperature for reaction for 16 hours; other procedures refer to example 1.
Example 14
This example used non-porous polystyrene divinylbenzene (particle size: 10 μm, non-porous, cat # 221610002) produced by Suzhou Seiki technology, Inc., which contained a large amount of epoxy groups on the surface.
Adding 60.0g of 65-70% ethylamine aqueous solution by weight percentage based on 100g of chromatographic medium with epoxy group on the surface, adding into a reaction device, and mechanically stirring and reacting for 16h at room temperature; other procedures refer to example 1.
Example 15
This example used non-porous polystyrene divinylbenzene (particle size: 10 μm, non-porous, cat # 221610002) produced by Suzhou Seiki technology, Inc., which contained a large amount of epoxy groups on the surface.
Adding 165.6g of methylamine water solution with the mass fraction of 30 percent into 100g of chromatographic medium with epoxy groups on the surface, adding the mixture into a reaction device, and mechanically stirring the mixture at room temperature for reaction for 16 hours; other procedures refer to example 1.
Example 16
Thioether diol example, epoxy grafting from media surface
In this example, porous polymethacrylate microspheres (particle size 30 μm, pore size 30 μm) produced by Suzhou Seiko technology Ltd were used
The goods number is: 281030952) having a large number of epoxy groups on the surface.
Adding 100ml of 0.1M sodium bicarbonate solution into 20.0g of chromatographic medium with epoxy groups on the surface, uniformly dispersing, adding 1.30g of 1-thioglycerol, adding into a reaction device, mechanically stirring at room temperature for reacting for 16 hours, and filtering and washing the medium to be neutral by deionized water after the reaction is finished.
Example 17
Example of Ether diol, epoxy grafting from the surface of the Medium
In this example, porous polymethacrylate microspheres (particle size 30 μm, pore size 30 μm) produced by Suzhou Seiko technology Ltd were used
Goods number: 281030952) having a large number of epoxy groups on the surface.
1) Adding 50ml of 5M sodium hydroxide aqueous solution into a chromatographic medium with 20.0g of epoxy groups on the surface, uniformly dispersing, adding into a reaction device, heating to 35 ℃, and keeping stirring for 2 hours.
2) Glycidol (20.0 g) was added and the reaction was stirred at 35 ℃ for 3 hours. Adding 100ml of purified water, stirring for 10min, and filtering the washing medium to be neutral by deionized water after the reaction is finished.
Example 18
Polyamine diol example, aldehyde group attachment from the media surface
In this example, porous polymethacrylate microspheres (particle size 30 μm, pore size 30 μm) produced by Suzhou Seiko technology Ltd were used
The goods number is: 280130950) having a large number of hydroxyl groups on the surface.
1) 100mL of 0.2M NaIO, based on 100.0g of chromatography medium having hydroxyl groups on the surface4And (3) mechanically stirring the solution at room temperature at 100rpm for reaction for 1.5h, and leaching and washing the microspheres by using deionized water with at least 10 times of volume after the reaction is finished.
2) Then 100mL of 0.1M phosphate buffer containing 1.52M dipropylene triamine (CAS #56-18-8, pH 7.2) was added along with 0.63g NaCNBH3And (3) mechanically stirring at 100rpm at room temperature for 5 hours for reaction, after the reaction is finished, repeatedly performing suction filtration and washing on the microspheres by using deionized water with at least 10 times of volume until the pH test paper of the filtrate is neutral, and performing suction drying until no filtrate is discharged.
3) The above medium was added to 150ml of a solution containing 50% glycidol and 2.0g of sodium hydrogencarbonate to complete the preparation of a chromatography medium according to step (4) in example 1.
Example 19
Polyamine diol example, aldehyde group attachment from the media surface
In this example, porous polymethacrylate microspheres (particle size 30 μm, pore size 30 μm) produced by Suzhou Seiko technology Ltd were used
The goods number is: 280130950) having a large number of hydroxyl groups on the surface.
1) 100mL of 0.2M NaIO, based on 100.0g of chromatography medium having hydroxyl groups on the surface4And (3) mechanically stirring the solution at room temperature at 100rpm for reaction for 1.5h, and leaching and washing the microspheres by using deionized water with at least 10 times of volume after the reaction is finished.
2) Then 100mL of 0.1M phosphate buffer containing 1.50M triethylenetetramine (CAS #112-24-3) was added, along with 0.63g of NaCNBH3And (3) mechanically stirring at 100rpm at room temperature for 5 hours for reaction, after the reaction is finished, repeatedly performing suction filtration and washing on the microspheres by using deionized water with at least 10 times of volume until the pH test paper of the filtrate is neutral, and performing suction drying until no filtrate is discharged.
3) The above medium was added to 150ml of a solution containing 50% glycidol and 2.0g of sodium hydrogencarbonate to complete the preparation of a chromatography medium according to step (4) in example 1.
Example 20
Amine mannose diol example, epoxy group attachment to media surface
In this example, porous polymethacrylate microspheres (particle size 30 μm, pore size 30 μm) produced by Suzhou Seiko technology Ltd were used
The goods number is: 281030952) having a large number of epoxy groups on the surface.
1) A chromatography medium having amine groups on the surface was prepared by following the steps (1) to (3) of example 1, based on 100g of the chromatography medium having epoxy groups on the surface.
2) 150ml of N, N-dimethylformamide solution containing 25% of alpha-D-mannopyranose chloride (2,3,4, 6-tetra-O-acetyl-alpha-D-mannopyranose chloride, CAS #14257-40-0) and 2.0g of sodium carbonate are added to the above medium, the mixture is mechanically stirred at room temperature and in the absence of light for reaction overnight, and after the reaction is finished, the medium is washed by suction filtration with at least 10 volumes of deionized water.
3) Adding 300ml of 0.5M sulfuric acid, adding the mixture into a reaction device, mechanically stirring the mixture for reaction for 2 hours at the temperature of 40 ℃, and filtering a washing medium by deionized water until the medium is neutral after the reaction is finished.
Example 21
Polyamine mannose diol example, aldehyde group attachment from the media surface
In this example, porous polymethacrylate microspheres (particle size 30 μm, pore size 30 μm) produced by Suzhou Seiko technology Ltd were used
The goods number is: 280130950) having a large number of hydroxyl groups on the surface.
1) 100mL of 0.2M NaIO, based on 100.0g of chromatography medium having hydroxyl groups on the surface4And (3) mechanically stirring the solution at room temperature at 100rpm for reaction for 1.5h, and leaching and washing the microspheres by using deionized water with at least 10 times of volume after the reaction is finished.
2) Then 100mL of 0.1M phosphate buffer containing 1.50M triethylenetetramine (CAS #112-24-3) was added, along with 0.63g of NaCNBH3And (3) mechanically stirring at 100rpm at room temperature for 5 hours for reaction, after the reaction is finished, repeatedly performing suction filtration and washing on the microspheres by using deionized water with at least 10 times of volume until the pH test paper of the filtrate is neutral, and performing suction drying until no filtrate is obtained, thereby preparing the chromatography medium with the polyamine on the surface.
3) 150ml of N, N-dimethylformamide solution containing 25% of alpha-D-mannopyranose chloride (2,3,4, 6-tetra-O-acetyl-alpha-D-mannopyranose chloride, CAS #14257-40-0) and 2.0g of sodium carbonate are added to the above medium, the mixture is mechanically stirred at room temperature and in the absence of light for reaction overnight, and after the reaction is finished, the medium is washed by suction filtration with at least 10 volumes of deionized water.
4) Adding 300ml of 0.5M sulfuric acid, adding the mixture into a reaction device, mechanically stirring the mixture for reaction for 2 hours at the temperature of 40 ℃, and filtering a washing medium by deionized water until the medium is neutral after the reaction is finished.
Example 22
Chromatographic medium separation of phenylboronic acid and ribonuclease A
The chromatographic media synthesized by the methods of examples 1 and 9 (comparative example) were packed into 4.6 x 50mm stainless steel chromatographic columns, respectively, and the adsorption-desorption of boron-containing molecules by the affinity chromatography media modified by the method of the present invention was compared with that of the conventional chromatography media with vicinal diols.
The samples selected were: phenylboronic acid (a boron-containing molecule); ribonuclease A (boron-free molecule)
The specific separation conditions were as follows:
sample preparation: ribonuclease A and phenylboronic acid mixed solution
Equilibrium solution: 0.25M ammonium acetate buffer (pH 8.0)
Elution solution: 0.1M formic acid solution
And (3) a separation process:
washing the equilibrium chromatographic column with an equilibrium solution;
loading a blank solvent, a single-mark ribonuclease A, a single-mark phenylboronic acid and a mixed-mark ribonuclease A and phenylboronic acid;
the balance solution is continuously washed until the baseline is stable;
eluting the phenylboronic acid adsorbed by the medium by using the elution solution;
washing the column with 0.5M NaOH;
(6) after use, the column was stored in 20% ethanol aqueous solution at room temperature.
Separation and purification experiment results:
example 1 the separation effect of affinity chromatography media (affinity chromatography media modified by the present invention) on phenylboronic acid and ribonuclease a is shown (fig. 3):
the chromatographic medium synthesized in example 1 has a strong adsorption effect on phenylboronic acid under alkaline conditions. The phenylboronic acid molecules are firmly combined with a chromatography medium, and the phenylboronic acid can be eluted under an acidic condition.
The chromatographic medium synthesized in example 1 has no effect on the adsorption of ribonuclease A under alkaline conditions. Ribonuclease A was carried directly out by the alkaline mobile phase.
The chromatographic medium synthesized in example 1 thus allows complete separation of phenylboronic acid from ribonuclease. The purity of the phenylboronic acid is more than 95 percent, and the yield is more than 90 percent.
(2) Example 9 (control) chromatography medium (normal chromatography medium with diol groups): the adsorption results for p-phenylboronic acid show (fig. 4):
the chromatographic medium synthesized in example 9 (comparative example) had an adsorptive effect on phenylboronic acid under alkaline conditions. But already starts to be eluted under alkaline conditions.
The chromatography media synthesized in example 9 (comparative example) adsorbed boron-containing molecules less strongly than the chromatography media of example 1.
Example 23:
chromatographic medium for separating boron-containing peptide molecules
The chromatographic media synthesized by the methods of examples 1 and 9 (comparative example) were packed into 4.6 x 50mm stainless steel chromatography columns, which were used to separate the target molecule from the crude product containing impurities. Testing the adsorption effect of the chromatography medium on the bortezomib molecules. The specific separation conditions were as follows:
sample preparation: sample solution containing bortezomib molecules
Loading solution: the pH value of the sample solution is adjusted to 8.0
Equilibrium solution: 0.25M ammonium acetate buffer (pH 8.0) + 20% methanol
Elution solution: 0.1M formic acid solution + 20% methanol
(1) Example 1 isolation and purification procedure:
washing and balancing the chromatographic column with a balancing solution;
sample solution loading;
the balance solution is continuously washed until the baseline is stable;
eluting the bortezomib molecules adsorbed by the medium by using the elution solution;
washing the column with 0.5M NaOH;
after use, the column was stored in 20% ethanol aqueous solution at room temperature.
Separation and purification experiment results:
example 1 the separation effect of the affinity chromatography media (affinity chromatography media modified according to the invention) on bortezomib molecules and their impurities is shown (fig. 5):
the chromatography medium synthesized in example 1 has a strong adsorption effect on bortezomib molecules under alkaline conditions. The bortezomib molecule is firmly combined with a chromatography medium, and can be eluted under an acidic condition.
The chromatographic medium synthesized in example 1 has no adsorption of impurities in most samples under alkaline conditions. Impurities are carried over directly by the alkaline mobile phase.
The chromatographic medium synthesized in example 1 can thus separate the bortezomib molecule and its impurities. The purity of bortezomib is more than 95 percent, and the yield is more than 90 percent.
(2) Example 9 (comparative example) isolation procedure:
washing the equilibrium chromatographic column with an equilibrium solution;
sample solution feeding;
the balance solution is continuously washed until the baseline is stable;
eluting the bortezomib molecules adsorbed by the medium by using the elution solution;
washing the column with 0.5M NaOH;
after use, the column was stored in 20% ethanol aqueous solution at room temperature.
Separation and purification experiment results:
example 9 (control) chromatography medium (normal chromatography medium with diol groups): results for bortezomib adsorption show (figure 6):
the chromatography media synthesized in example 9 (comparative example) had an adsorption effect on bortezomib under basic conditions. But already starts to be eluted under alkaline conditions.
The chromatography medium synthesized in example 9 (comparative example) had weaker adsorption of bortezomib than the chromatography medium of example 1.
(3) Analysis of results of bortezomib separation and purification experiments
The chromatographic medium in example 1 has multiple synergistic effects to make the o-polyhydroxy group combine with boron atom more easily in space and conformation to form effective coordination, and the purity and yield of bortezomib are high;
example 9 (comparative example) the chromatography medium surface had only a normal cis-diol group, which was not easily bonded to the boron atom to form an effective ligand, and the results of the separation and purification experiments were inferior to those of example 1.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (13)
1. A chromatographic medium for separating boron-containing species, wherein the chromatographic medium comprises a base support and grafted chains grafted to the base support;
the general formula of the grafting chain is MA-R1-X1-R2Or
Shown;
wherein MA is a group that can be linked to other chemical functional groups, either covalently or non-covalently;
R1、R2and R3Independently selected from hydrogen atoms, saturated hydrocarbon groups, unsaturated hydrocarbon groups or saturated hydrocarbon groups containing heteroatoms which are not carbon electronegative, and at least one of the groups carries cis-ortho-polyhydroxy, wherein the cis-ortho-polyhydroxy is a linear chain skeleton structure or a ring chain skeleton structure;
X1and X2Divalent and trivalent heteroatoms of said non-carbon electronegative heteroatom, respectively; r in the graft chain1、R2And R3Sum of number of carbon atoms present therein>2 pieces of the Chinese herbal medicines.
2. The chromatographic medium according to claim 1, wherein the non-carbon electronegative heteroatoms each have a paulin electronegative number greater than 2.0;
preferably, the non-carbon electronegative heteroatom is selected from one or more of O, S, N, P;
preferably, the saturated hydrocarbon group containing a non-carbon electronegative heteroatom may be polyamine, oligoimine, polyether, oligoether, polythioether, oligothioether; more preferred are polyamines and oligoimines, such as: ethylenediamine, diethylenetriamine, dipropylenetriamine, triethylenetetramine, linear and branched oligoethyleneimines.
3. Chromatography media according to claim 2, wherein X is1Selected from O or S atoms.
4. Chromatography media according to claim 2, wherein X is2Selected from N or P atoms.
5. The chromatography media of claim 1, wherein MA is selected from at least one of epoxy, amino, aldehyde, hydroxyl, carboxyl, oxo, and thiol groups.
6. The chromatography media of claim 1, wherein R is1、R2And R3At least one of which carries a cis-ortho-polyol; preferably a cis-vicinal diol; more preferably a terminal cis-vicinal diol;
preferably R1、R2And R3At least one of which carries a catechol, such as a catechol homolog;
the ring chain framework structure is preferably monosaccharide, disaccharide or polysaccharide; the backbone structure of the ring chain is preferably mannose, fructose, galactose, mannose-containing disaccharides and mannose-containing oligosaccharides.
7. The chromatography media of any one of claims 1 to 6, wherein the matrix support comprises any one of borosilicate glass, agar, agarose derivatives, magnetic beads, silica, titanium dioxide, alginate, cellulose derivatives, dextran, starch, cyclodextrin, chitosan, carrageenan, guar gum, gum arabic, gum ghatti, gum tragacanth, karaya gum, locust bean gum, xanthan gum, pectin, mucin, liver thioesters and gelatin, silica, ceramic, glass, polyurethane, polystyrene divinyl benzene, polymethylmethacrylate, polyacrylamide, polyethylene glycol terephthalate, polyvinyl acetate, polyethylene, polypropylene, polyvinyl chloride, polyvinyl pyrrolidone, or copolymers of any of the foregoing.
8. A chromatography medium according to any one of claims 1 to 6, wherein the matrix support is in particulate form, having a particle size in the range of from 1 μm to 500 μm, preferably from 1 μm to 300 μm; more preferably 2 to 200. mu.m.
9. A chromatography media according to any of claims 1 to 6, wherein the matrix support is non-porous or comprises one or more pores; preferably having a pore diameter of
More preferably the pore diameter is
It is further preferred that the pore diameter is
10. A chromatographic separation device comprising the chromatographic medium according to any one of claims 1 to 9.
11. Use of a chromatographic medium as claimed in any one of claims 1 to 9, or a chromatographic separation device as claimed in claim 10, for separating or enriching a boron-containing material from a liquid medium.
12. Use according to claim 11, characterized in that the chromatography medium adsorbs the boron containing substance under alkaline conditions, elutes the boron containing substance under acidic conditions;
preferably, the mobile phase for eluting the boron-containing substance contains a carbohydrate; the carbohydrate is preferably sorbitol.
13. Use according to claim 11 or 12, wherein the boron-containing substance comprises the following molecule:
a) boron-containing compounds for proteasome inhibitors, such as bortezomib, efavir;
b) boron-containing, anti-fungal, bacterial, viral, organoboronic drug molecules such as tavaborole, kreb's and vaborbacam-Meropenem;
c) boron compound carriers, such as boracic acridine derivatives, for use in neutron capture tumor therapy;
d) a boron-containing compound having thrombin inhibitory activity;
e) boron-containing oligonucleotides, polypeptides, proteins having therapeutic effects;
f) boron compounds for use in boron selective reactions for molecular modular synthesis;
g) for the recovery of unreacted or excess boron-containing reagent in a chemical reaction, preferably for solid surface or polymer and soluble boron-containing reagent reactions.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115166073A (en) * | 2022-06-21 | 2022-10-11 | 北京泰德制药股份有限公司 | Method for detecting ixazofamid or salt isomer thereof |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998005627A1 (en) * | 1996-08-05 | 1998-02-12 | Prolinx, Inc. | Boronic compound complexing reagents and highly stable complexes |
| JP2004089824A (en) * | 2002-08-30 | 2004-03-25 | Sangaku Renkei Kiko Kyushu:Kk | Boron bonding agent |
| US20100101996A1 (en) * | 2007-01-30 | 2010-04-29 | Kowa Company, Ltd. | Boron-removing agent and boron removal method utilizing xerogel |
| CN102123789A (en) * | 2008-06-26 | 2011-07-13 | 3M创新有限公司 | Solid support with a grafted chain |
| CN102219800A (en) * | 2011-04-20 | 2011-10-19 | 南京大学 | Organic substituted boric acid ester, boron affinity functional material using organic substituted boric acid ester as functional monomer as well as preparation and application of organic substituted boric acid ester |
| JP2012046596A (en) * | 2010-08-25 | 2012-03-08 | Tokyo Metropolitan Univ | Polymer gel, method for producing the same, treating agent for cleaning water, and treating method for cleaning water |
| CN104549182A (en) * | 2014-12-29 | 2015-04-29 | 上海烟草集团有限责任公司 | Branched boric acid functionalized monolithic column and preparation method and application thereof |
| CN104624171A (en) * | 2014-12-30 | 2015-05-20 | 浙江大学 | Polymer grafting type hydrophobic charge-induced chromatography medium and preparation method thereof |
| CN109206614A (en) * | 2017-07-05 | 2019-01-15 | 大连医科大学 | A kind of reagent and its preparation method and application of the detection containing sugar chain biomolecule |
-
2020
- 2020-06-09 CN CN202010526191.0A patent/CN111729661B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998005627A1 (en) * | 1996-08-05 | 1998-02-12 | Prolinx, Inc. | Boronic compound complexing reagents and highly stable complexes |
| JP2004089824A (en) * | 2002-08-30 | 2004-03-25 | Sangaku Renkei Kiko Kyushu:Kk | Boron bonding agent |
| US20100101996A1 (en) * | 2007-01-30 | 2010-04-29 | Kowa Company, Ltd. | Boron-removing agent and boron removal method utilizing xerogel |
| CN102123789A (en) * | 2008-06-26 | 2011-07-13 | 3M创新有限公司 | Solid support with a grafted chain |
| JP2012046596A (en) * | 2010-08-25 | 2012-03-08 | Tokyo Metropolitan Univ | Polymer gel, method for producing the same, treating agent for cleaning water, and treating method for cleaning water |
| CN102219800A (en) * | 2011-04-20 | 2011-10-19 | 南京大学 | Organic substituted boric acid ester, boron affinity functional material using organic substituted boric acid ester as functional monomer as well as preparation and application of organic substituted boric acid ester |
| CN104549182A (en) * | 2014-12-29 | 2015-04-29 | 上海烟草集团有限责任公司 | Branched boric acid functionalized monolithic column and preparation method and application thereof |
| CN104624171A (en) * | 2014-12-30 | 2015-05-20 | 浙江大学 | Polymer grafting type hydrophobic charge-induced chromatography medium and preparation method thereof |
| CN109206614A (en) * | 2017-07-05 | 2019-01-15 | 大连医科大学 | A kind of reagent and its preparation method and application of the detection containing sugar chain biomolecule |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115166073A (en) * | 2022-06-21 | 2022-10-11 | 北京泰德制药股份有限公司 | Method for detecting ixazofamid or salt isomer thereof |
| CN115166073B (en) * | 2022-06-21 | 2023-06-13 | 北京泰德制药股份有限公司 | Method for detecting isomer of I Sha Zuomi or salt thereof |
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