CN108043375A - A kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases - Google Patents
A kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases Download PDFInfo
- Publication number
- CN108043375A CN108043375A CN201711086731.2A CN201711086731A CN108043375A CN 108043375 A CN108043375 A CN 108043375A CN 201711086731 A CN201711086731 A CN 201711086731A CN 108043375 A CN108043375 A CN 108043375A
- Authority
- CN
- China
- Prior art keywords
- vancomycin
- silicon ball
- preparation
- mode combination
- stationary phases
- 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.)
- Pending
Links
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
- B01J20/29—Chiral phases
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- 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
- B01D15/3833—Chiral chromatography
Landscapes
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Peptides Or Proteins (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The present invention relates to a kind of preparation methods of multi-mode combination vancomycin chromatographic stationary phases, amino is modified in silicon ball surface, then using glutaraldehyde as spacerarm, it is reacted by aldimine condensation, big cyclohexanol peptide antibiotic vancomycin is bonded to silicon ball surface, bonding termination process is carried out using the amino acid or sugar of small molecule again, prepares multi-mode combination chiral chromatogram stationary phase；There are a variety of functional groups in vancomycin structure, the lock out operation being suitable under different chromatographic isolation patterns, in addition, spacerarm is made with glutaraldehyde, big cyclohexanol peptide antibiotics is bonded to silicon ball surface, effectively reduce the influence of space steric effect, improve bonding efficiency, and the termination process of small molecule can either shield negative effect of the remaining silicone hydroxyl in chromatographic separation process, or separation provides more and quantity action site, improves separative efficiency, improves separation selectivity.
Description
Technical field
The present invention relates to technical field of textile production, are that a kind of multi-mode combination vancomycin chromatography is fixed specifically
The preparation method of phase.
Background technology
Vancomycin has a variety of functional groups, 3 cavitys, 18 is included in structure as a kind of big anti-glycopeptide life element of ring
A chiral centre, carboxyl, amide group, glucosides group etc., some researches show that the stationary phase of vancomycin bonding modification is suitable
Chiral resolution under a variety of chromatography clastotypes such as reverse phase, positive, hydrophilic, polarity-organic phase.
Since the complexity of its mechanism of action is various, vancomycin bonded stationary phase can be realized to the complicated body such as biological sample
The separation of system has very extensive application potential, can be applied to the chromatographic isolation of the components such as protein, polypeptide, it can also be used to
The fields such as the separation of Chinese medicine system, the fractionation of chipal compounds.
The currently used technology that vancomycin is bonded to Silica Surface is by directly by the silica gel containing epoxide group
Modifying agent is bonded to silicon ball surface, and then using the reaction of epoxy group and amino, vancomycin is bonded to Silica Surface.It is this
The advantage of method is that synthesis step is simple, and operating procedure is few, advantage of lower cost, but since macrocyclic antibiotic molecule is larger,
Steric effect is apparent in bonding process, and silicon ball surface has substantial amounts of silicone hydroxyl remaining, this can cause chromatographic separation process sternly
The negative effect (such as chromatographic peak hangover, broadening) of weight.
The content of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide a kind of multi-mode combination vancomycin chromatographies to fix
The preparation method of phase.
The purpose of the present invention is what is be achieved through the following technical solutions：
A kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases, concretely comprises the following steps：
(1) activation of silica gel microball and silanization
Silica gel microball is taken, is dispersed in the dilute alkaline soln of 0.05mol/L, 25~35min of ultrasound is washed with water to neutrality,
After drying, silicon ball is dispersed in methanol-water solution, addition 3-aminopropyltriethoxysilane APTES, after ultrasonic mixing,
Heating reflux reaction 20~for 24 hours, methanol cleaning is dry, obtains amido modified silicon ball；
In step (1), the silica gel microball is mesoporous silicon sphere；
In step (1), the dilute alkaline soln is sodium hydroxide or potassium hydroxide aqueous solution；
In step (1), the ratio of handled silica gel microball and the dilute alkaline soln that need to be used is 1:8~1:10(g/
mL)；
In step (1), the volume ratio of methanol-water is 7:1~8:1；
In step (1), the ratio of used silicon ball and the methanol-water solution that need to be used is 1:10~1:12(g/
ML), the mass ratio of silicon ball and the dosage of APTES reagents is 1:1.5~1:2(g/g)；
In step (1), described is heated to reflux temperature as 65~75 DEG C.
(2) the vancomycin modification of silica gel microball
Silicon ball after silanization treatment is dispersed in the glutaraldehyde water solution that mass concentration is 10%~15%, ultrasonic mixing
Afterwards, 4~6h of heating reflux reaction, methanol cleaning is dry, obtains aldehyde group modified silicon ball；By aldehyde radicalization treated silicon ball point
It dissipates in aqueous solution, adds in the vancomycin aqueous solution of 10g/L, after ultrasonic mixing, heating reflux reaction 4~6h is abundant with water
Cleaning, filtering obtain the silicon ball of vancomycin modification；
In step (2), the mass percent of used glutaraldehyde water solution is 10%~15%；
In step (2), the silicon ball of used silanization treatment and the usage ratio of glutaraldehyde water solution are 1:10~
1:12(g/mL)；
In step (2), described is heated to reflux temperature as 55~65 DEG C；
In step (2), the usage ratio of the used processed silicon ball of aldehyde radicalization and vancomycin aqueous solution is 1:8
~1:10(g/mL)；
(3) small molecule termination process
Silicon ball after vancomycin is modified is dispersed in micromolecular compound aqueous solution, 10~12h of heating reflux reaction, after
It is cleaned up, dried with pure water；
In step (3), used micromolecular compound is the small molecules chemical combination such as the amino acid containing amino or carboxyl, sugar
Object；
In step (3), the concentration of used micromolecular compound aqueous solution is 0.05~0.1mol/L；
In step (3), the usage ratio of used silicon ball and micromolecular compound aqueous solution is 1:15~1:20(g/
mL)；
In step (3), described is heated to reflux temperature as 50~60 DEG C.
(4) reduction of carbon-to-nitrogen double bon
Silicon ball after vancomycin is modified and blocked is dispersed in the aqueous dibasic potassium phosphate solution of 0.02mol/L, is added in
NaBH4, be stirred to react 2~3h at room temperature, filter, after cleaned with pure water to neutrality, drying；
In step (4), the concentration of used aqueous dibasic potassium phosphate solution is 0.02mol/L；
In step (4), the usage ratio of used silicon ball and aqueous dibasic potassium phosphate solution should be 1:10~1:12(g/
ML), used silane and the NaBH used4Usage ratio be 1:0.1~1:0.2(g/g).
Compared with prior art, the positive effect of the present invention is：
(1) in vancomycin structure there are a variety of functional groups, a variety of interactions can be provided, therefore vancomycin bonding is repaiied
The silicon ball of decorations is suitable for the chiral resolution under a variety of chromatography clastotypes such as reverse phase, positive, hydrophilic, polarity-organic phase；
(2) spacerarm is made with glutaraldehyde and big cyclohexanol peptide antibiotics is bonded to silicon ball surface, effectively reduce big point
The influence of sub- steric effect can greatly improve bonding efficiency；
(3) big cyclohexanol peptide antibiotics has a variety of functional groups in itself, is capable of providing a variety of interactions, ensure that fixation
The chiral resolution ability of phase；
(4) special sealing processing is carried out with small molecular sugar or amino acid, effectively shields the influence of silicone hydroxyl, and can carry
For additional interaction, the separating capacity of chromatography separation media is further enhanced.
Description of the drawings
Attached drawing 1 is the molecular structural formula schematic diagram of vancomycin；
Attached drawing 2 is that the preparation process using glutaraldehyde as the vancomycin bonding multi-mode chiral chromatogram stationary phase of spacerarm is shown
It is intended to；
Fig. 3 be using vancomycin bonded stationary phase under rp mode to the separation chromatography of hydroquinone and catechol
Figure, chromatographic condition：Mobile phase (A) water, (B) methanol；Isocratic 65%A；Detection wavelength 254nm；Flow velocity 0.8mL/min.
Specific embodiment
A kind of specific embodiment party of the preparation method of multi-mode combination vancomycin chromatographic stationary phases of the present invention presented below
Formula.
Embodiment 1
According to method shown in attached drawing 2, in Silica Surface using glutaraldehyde as spacerarm, vancomycin is bonded, prepares multi-mode
Chiral chromatogram stationary phase, specific preparation process are：
(1) pretreatment of silica gel microball
Take 20g silica gel microballs (5 μm of particle diameter, aperture), it is dispersed in the sodium hydroxide solution of 0.1mol/L,
After ultrasonic 30min, it is 6.5~7 that dilute hydrochloric acid is added, which to adjust pH value, after fully being cleaned with water, is cleaned three times, 50 DEG C with methanol or acetonitrile
Lower drying, it is spare.
(2) silanization treatment of silica gel microball
The silica gel particle of activation in step (1) is dispersed in 80mL methanol-waters (volume ratio 7:1) in solution, add successively
Enter 20mL APTES reagents, after ultrasonic mixing, for 24 hours, methanol cleans 3 times back flow reaction under the conditions of 65 DEG C, dry, obtains amino and repaiies
The silicon ball of decorations.
(3) aldehyde radical of silica gel microball
Silicon ball after silanization treatment is disperseed in aqueous solution, to add in 100mL glutaraldehyde water solution (mass concentrations
10%), after ultrasonic mixing, 3~4h of heating reflux reaction at 55 DEG C, methanol cleaning is dry, obtains aldehyde group modified silicon ball；
(4) the vancomycin modification of silica gel microball and small molecule sealing end
By aldehyde radicalization treated silicon ball disperses in aqueous solution, add in 100mL vancomycins aqueous solution (42g/L), surpass
After sound mixing, 3~4h of heating reflux reaction at 60 DEG C is rear to add in 30mL amino acid solutions (0.01mol/L), and it is anti-to continue heating
Answer 2h, after cleaned up with pure water, dry.
(5) reduction of carbon-to-nitrogen double bon
Silicon ball after vancomycin is modified and blocked is dispersed in 100mL aqueous dibasic potassium phosphate solutions (0.02mol/L),
Add in 2g NaBH4, be stirred to react 2~3h at room temperature, filter, after cleaned with pure water to neutrality, drying.
(6) chromatography characterizes
Prepared chromatographic stationary phases are applied to the separation of benzenediol isomer, as shown in Figure 3, result table
Bright, prepared chromatographic stationary phases have good separating capacity for isomer.
A kind of new multi-mode using vancomycin as functional group of the present invention combines the preparation of chiral chromatogram stationary phase
Method modifies amino in silicon ball surface, then using glutaraldehyde as spacerarm, is reacted by aldimine condensation, big cyclohexanol peptide is resisted
Raw element vancomycin is bonded to silicon ball surface, then carries out bonding termination process using the amino acid or sugar of small molecule, prepares more
Mode combinations chiral chromatogram stationary phase；In vancomycin structure there are a variety of functional groups, be suitable under different chromatographic isolation patterns
Lock out operation, in addition, make spacerarm with glutaraldehyde is bonded to silicon ball surface by big cyclohexanol peptide antibiotics, effectively reduce
The influence of space steric effect improves bonding efficiency, and the termination process of small molecule can either shield remaining silicone hydroxyl in chromatography
Negative effect in separation process, or separation provides more and quantity action site, improves separative efficiency, changes
Kind separation selectivity.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
Member, without departing from the inventive concept of the premise, can also make several improvements and modifications, these improvements and modifications also should be regarded as
In protection scope of the present invention.
Claims (10)
1. a kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases, which is characterized in that it is concretely comprised the following steps：
(1) activation of silica gel microball and silanization
Silica gel microball is taken, is dispersed in the dilute alkaline soln of 0.05mol/L, 25~35min of ultrasound is washed with water to neutrality, drying
Afterwards, silicon ball is dispersed in methanol-water solution, adds in 3-aminopropyltriethoxysilane (APTES reagents), ultrasonic mixing
Afterwards, heating reflux reaction 20~for 24 hours, methanol cleaning is dry, obtains amido modified silicon ball；
(2) the vancomycin modification of silica gel microball
Silicon ball after silanization treatment is dispersed in mass concentration as 10%~15% glutaraldehyde water solution, after ultrasonic mixing,
4~6h of heating reflux reaction, methanol cleaning is dry, obtains aldehyde group modified silicon ball；By aldehyde radicalization, treated that silicon ball is dispersed in
In aqueous solution, the vancomycin aqueous solution of 10g/L is added in, after ultrasonic mixing, 4~6h of heating reflux reaction is fully cleaned with water,
Filtering obtains the silicon ball of vancomycin modification；
(3) small molecule termination process
Silicon ball after vancomycin is modified is dispersed in micromolecular compound aqueous solution, 10~12h of heating reflux reaction, afterwards with pure
Water cleans up, drying；
(4) reduction of carbon-to-nitrogen double bon
Silicon ball after vancomycin is modified and blocked is dispersed in the aqueous dibasic potassium phosphate solution of 0.02mol/L, adds in NaBH4,
Be stirred to react 2~3h at room temperature, filter, after cleaned with pure water to neutrality, drying.
2. a kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases as described in claim 1, which is characterized in that
In step (1), the silica gel microball is mesoporous silicon sphere；
In step (1), the dilute alkaline soln is sodium hydroxide or potassium hydroxide aqueous solution；
In step (1), the ratio of handled silica gel microball and the dilute alkaline soln that need to be used is 1:8~1:10(g/mL).
3. a kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases as described in claim 1, which is characterized in that
In step (1), the volume ratio of methanol-water is 7:1~8:1；
In step (1), the ratio of used silicon ball and the methanol-water solution that need to be used is 1:10~1:12 (g/mL), silicon
Ball and the mass ratio of the dosage of APTES reagents are 1:1.5~1:2(g/g)；
In step (1), described is heated to reflux temperature as 65~75 DEG C.
4. a kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases as described in claim 1, which is characterized in that
In step (2), the mass percent of used glutaraldehyde water solution is 10%~15%；
In step (2), the silicon ball of used silanization treatment and the usage ratio of glutaraldehyde water solution are 1:10~1:12
(g/mL)。
5. a kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases as described in claim 1, which is characterized in that
In step (2), described is heated to reflux temperature as 55~65 DEG C；
In step (2), the usage ratio of the used processed silicon ball of aldehyde radicalization and vancomycin aqueous solution is 1:8~1:
10(g/mL)。
6. a kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases as described in claim 1, which is characterized in that
In step (3), used micromolecular compound is the micromolecular compounds such as the amino acid containing amino or carboxyl, sugar；
In step (3), the concentration of used micromolecular compound aqueous solution is 0.05~0.1mol/L.
7. a kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases as described in claim 1, which is characterized in that
In step (3), the usage ratio of used silicon ball and micromolecular compound aqueous solution is 1:15~1:20(g/mL).
8. a kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases as described in claim 1, which is characterized in that
In step (3), described is heated to reflux temperature as 50~60 DEG C.
9. a kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases as described in claim 1, which is characterized in that
In step (4), the concentration of used aqueous dibasic potassium phosphate solution is 0.02mol/L.
10. a kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases as described in claim 1, feature exist
In,
In step (4), the usage ratio of used silicon ball and aqueous dibasic potassium phosphate solution should be 1:10~1:12 (g/mL),
Used silane and the NaBH used4Usage ratio be 1:0.1~1:0.2(g/g).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711086731.2A CN108043375A (en) | 2017-11-07 | 2017-11-07 | A kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711086731.2A CN108043375A (en) | 2017-11-07 | 2017-11-07 | A kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108043375A true CN108043375A (en) | 2018-05-18 |
Family
ID=62118966
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711086731.2A Pending CN108043375A (en) | 2017-11-07 | 2017-11-07 | A kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108043375A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110876925A (en) * | 2018-09-05 | 2020-03-13 | 武汉大学 | Mixed-mode chromatographic stationary phase and preparation method and application thereof |
| CN111359263A (en) * | 2020-02-18 | 2020-07-03 | 青岛大学 | Capillary coated column for chiral compound separation, preparation method and application thereof |
| CN113332967A (en) * | 2021-06-29 | 2021-09-03 | 常州工程职业技术学院 | Trapezoidal polyether modified and cysteine terminated chromatographic stationary phase, preparation method and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004258013A (en) * | 2003-02-28 | 2004-09-16 | Shiseido Co Ltd | Filler for chromatography |
| CN1772371A (en) * | 2005-10-14 | 2006-05-17 | 浙江大学 | Chiral chromatographic fixed phase stuffing of vancomycin phenylisocyanate and its prepn |
-
2017
- 2017-11-07 CN CN201711086731.2A patent/CN108043375A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2004258013A (en) * | 2003-02-28 | 2004-09-16 | Shiseido Co Ltd | Filler for chromatography |
| CN1772371A (en) * | 2005-10-14 | 2006-05-17 | 浙江大学 | Chiral chromatographic fixed phase stuffing of vancomycin phenylisocyanate and its prepn |
Non-Patent Citations (1)
| Title |
|---|
| 雷雯等: ""以戊二醛为间隔臂的万古霉素键合手性固定相制备"", 《分析化学研究报告》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110876925A (en) * | 2018-09-05 | 2020-03-13 | 武汉大学 | Mixed-mode chromatographic stationary phase and preparation method and application thereof |
| CN110876925B (en) * | 2018-09-05 | 2022-04-12 | 武汉大学 | Mixed-mode chromatographic stationary phase and preparation method and application thereof |
| CN111359263A (en) * | 2020-02-18 | 2020-07-03 | 青岛大学 | Capillary coated column for chiral compound separation, preparation method and application thereof |
| CN113332967A (en) * | 2021-06-29 | 2021-09-03 | 常州工程职业技术学院 | Trapezoidal polyether modified and cysteine terminated chromatographic stationary phase, preparation method and application |
| CN113332967B (en) * | 2021-06-29 | 2023-04-07 | 常州工程职业技术学院 | Trapezoidal polyether modified and cysteine terminated chromatographic stationary phase, preparation method and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108043375A (en) | A kind of preparation method of multi-mode combination vancomycin chromatographic stationary phases | |
| CN105548538A (en) | IgM antibody combined detection device of Coxsackie virus A16 and enterovirus 71 and making method thereof | |
| CN107262078B (en) | Graphene/silica gel solid phase extraction material and application thereof | |
| CN102558463B (en) | One-pot synthesis of phenylboronic acid polymer magnetic nano composite material and preparation method and application thereof | |
| CN106861661B (en) | Monosaccharide polymer enrichment material and its preparation and the application in glycopeptide enrichment | |
| CN109174009B (en) | Aptamer modified triazine covalent organic framework composite material, preparation method and application | |
| CN108276584A (en) | The detection method of aromatic amine compound in a kind of human urine | |
| EP3369479B1 (en) | Adsorbing material for multiple pathogenic factors of sepsis as well as preparation method and application thereof | |
| KR20110028295A (en) | Method for preparation of sugar chain sample, sugar chain sample, and method for analysis of sugar chain | |
| US10758890B2 (en) | Substrate for the purification of biological liquids | |
| CN103055832B (en) | Chromatographic packing for separating water soluble polymer and protein and preparation method of same | |
| CN104001481A (en) | Preparation method for hydrophilic magnetic nano material for enrichment of glycopeptides | |
| CN101513577A (en) | Melamine molecular imprinted solid phase extraction cartridge and a preparation method and application thereof | |
| CN109621912A (en) | A kind of coating method of blood perfusion acticarbon | |
| CN108855003A (en) | It is a kind of for removing the immunosorbent and preparation method thereof of inflammatory factor in blood | |
| CN108514864A (en) | A kind of chitin/graphene oxide composite sponge and the preparation method and application thereof | |
| JPH0338891B2 (en) | ||
| CN105954404A (en) | Method for determining content of salivary acid in serum by using UIO-66-NH2 material | |
| CN109647359B (en) | Preparation and application of ionic liquid functionalized polyacrylonitrile nanofiber membrane | |
| CN111239314B (en) | Separation and analysis method of chitin oligosaccharide | |
| CN108059673B (en) | Method for separating immunoglobulin IgG from human serum | |
| CN101942107B (en) | Preparation method of protein-printed polymer of carboxymethyl chitosan and chitosan | |
| CN107999035A (en) | Chromatography media using tryptamines as functional ligand | |
| CN112645994B (en) | Extraction process of salidroside | |
| CN108707201A (en) | A kind of Arab's gala oligomeric polysaccharide and its preparation and application |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180518 |