CN104324707A - Amino diacid hydrophilic chromatographic stationary phase and preparation method thereof - Google Patents
Amino diacid hydrophilic chromatographic stationary phase and preparation method thereof Download PDFInfo
- Publication number
- CN104324707A CN104324707A CN201410471342.1A CN201410471342A CN104324707A CN 104324707 A CN104324707 A CN 104324707A CN 201410471342 A CN201410471342 A CN 201410471342A CN 104324707 A CN104324707 A CN 104324707A
- Authority
- CN
- China
- Prior art keywords
- silica gel
- stationary phase
- chromatographic stationary
- obtains
- dibasic aminoacid
- 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
- B01J20/286—Phases chemically bonded to a substrate, e.g. to silica or to polymers
- B01J20/288—Polar phases
-
- 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
- B01J20/282—Porous sorbents
- B01J20/283—Porous sorbents based on silica
-
- 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
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/3085—Chemical treatments not covered by groups B01J20/3007 - B01J20/3078
-
- 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
- B01J2220/00—Aspects relating to sorbent materials
- B01J2220/50—Aspects relating to the use of sorbent or filter aid materials
- B01J2220/52—Sorbents specially adapted for preparative chromatography
Landscapes
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The invention discloses an amino diacid hydrophilic chromatographic stationary phase, the structure of which is defined in the specification. A preparation method of the amino diacid hydrophilic chromatographic stationary phase comprises the following steps: (1) a terminal alkynyl group is introduced to the surface of silica gel: a, a silane coupling agent and propargylamine are added into an organic solvent; b, activated microspheric silica gel is added; c, reaction, filtration and washing are carried out to obtain a solid; and d, vacuum drying is carried out to obtain terminal alkynyl silica gel; and (2) the amino diacid hydrophilic chromatographic stationary phase is bonded by click chemistry: a, the alkynyl group on the silica gel is prepared; b, amino diacid modified by an azide group and a catalyst are added; c, reaction, filtration and washing are carried out to obtain a solid; and d, vacuum drying is carried out to obtain the amino diacid hydrophilic chromatographic stationary phase. As click chemistry is adopted as a bonding reaction method, immobilization with high selectivity and high conversion rate can be realized under a mild condition. The preparation process is simple, and the structure of each intermediate is clear. The prepared amino diacid hydrophilic chromatographic stationary phase, used as a polar stationary phase, can separate strongly polar compounds efficiently in the mode of hydrophilic liquid chromatogram.
Description
Technical field
The present invention relates to Stationary Phase for HPLC technology, specifically, is that a kind of " click chemistry " (Click chemistry) that use is as dibasic aminoacid hydrophilic chromatographic stationary phase of bonding method bonding and preparation method thereof.
Technical background
Hydrophilic Interaction Chromatography (HILIC) is the chromatographic technique for separating of strong polar compound that development in recent years is got up, and is a kind of with polar stationary phase (structural modification as silica gel or Silica Surface) and be the chromatogram mode of mobile phase containing high concentration polar organic solvent and low concentration aqueous solution.Hydrophilic Interaction Chromatography solves conventional inverter chromatogram hydrophobicity Stationary liquid (as C18, C8 etc.) and retains very weak to strong polar compound (as nucleosides, cephalo, training south etc.), even uncensored problem.Hydrophilic Interaction Chromatography is for separating of the existing bibliographical information of polar compound (as amino acid, peptide and sugar etc.).The Stationary liquid of hydrophilic Interaction Chromatography is still use positive Stationary liquid, mainly directly use silica gel, amino, amide groups, glycol-based, carbohydrate etc. as Stationary liquid, but, dissimilar Stationary liquid also exists different retention mechanisms and separating property, wherein, there are some larger shortcomings in the Stationary liquid of some type, such as: Irreversible Adsorption phenomenon easily occurs silicagel column; Nh 2 column can generate Schiff with carbohydrate; Thus impact is separated.At present, the Stationary liquid specially for hydrophilic Interaction Chromatography exploitation is at the early-stage, and the commercial hydrophilic Interaction Chromatography Stationary liquid kind of energy is less, thus needs the Stationary liquid developing other types to meet the demand for development of hydrophilic Interaction Chromatography.
" click chemistry " (Click chemistry), also " click chemistry ", " dynamic combinatorial chemistry " or " quick-acting conjunction combined chemical " is translated into, be the synthesis concept proposed by people such as chemist's Barry summer Prices (K.B.Sharpless), its core is: obtain molecular diversity widely with simple and reliable on a small quantity with chemical transformation that is high selectivity." click chemistry " is started fast, effectively or even 100% reliably, highly selective manufactures the synthetic chemistry frontier of all kinds of noval chemical compound, wherein, the reaction of extensive use is Huisgen 1 the most, 3-diploar cycloaddition reacts, it has played very important effect in the synthesis of chromatographic stationary phases, and its reaction equation is:
。
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, there is provided a kind of dibasic aminoacid hydrophilic chromatographic stationary phase, adopt dibasic aminoacid as polar functional group, its character is very stable, its surface texture can not change because of the change of pH, is also not easy to react with solute molecule; The efficient separation to strong polar compound can be realized under hydrophilic interaction liquid chromatogram pattern as polar stationary phase.The present invention's second object provides the preparation method of described hydrophilic chromatographic stationary phase, adopts " click chemistry " as bonding reaction method, can realize the immobilized of high selectivity and high conversion in a mild condition; Preparation process is simple, and each intermediate structure is clear and definite, can effectively prepare described hydrophilic chromatographic stationary phase.
For achieving the above object, present invention employs following technical scheme.
A kind of dibasic aminoacid hydrophilic chromatographic stationary phase, it is characterized in that, its structure is:
。
A preparation method for dibasic aminoacid hydrophilic chromatographic stationary phase, uses " click chemistry " to be bonding method bonding dibasic aminoacid molecule, comprises the following steps:
(1) Silica Surface introduces terminal acetylene
1. add silane coupler, propargylamine in organic solvent, the mol ratio of silane coupler and propargylamine is 1 ~ 5:1.2 ~ 5, and needed for every gram of silica gel, the amount of organic solvent is 5 ~ 30mL, reacts 6 ~ 36 hours under 60 ~ 150 DEG C of conditions;
2. add the micro-spherical silica gel of activation, the silane coupler needed for every gram of micro-spherical silica gel is 1 ~ 10mmol;
3. under 80 ~ 120 DEG C of conditions, continue reaction 12 ~ 48 hours, then filter with sand core funnel, more successively with the washing of carrene, methyl alcohol and acetone, obtain solids;
4. the solids that step (1) 3. obtains is placed in vacuum drying chamber under 40 ~ 80 DEG C of conditions dry 6 ~ 12 hours, obtains terminal acetylene silica gel;
It is characterized in that:
(2) by " click chemistry " bonding dibasic aminoacid hydrophilic chromatographic stationary phase
1. the terminal acetylene silica gel that step (1) 4. obtains being added volume ratio is that in the water of 2 ~ 10 ︰ 1 and the mixed solvent of methyl alcohol, needed for every gram of alkynyl silica gel, the amount of water and methanol mixed solvent is 20 ~ 50mL, obtains alkynyl on silica gel;
2. the dibasic aminoacid and catalyst that are modified with azido group is added: the molar dose that the dibasic aminoacid (nitrine dibasic aminoacid) being modified with azido group adds is 1 ~ 15 times of the molar dose of alkynyl on the silica gel that 1. obtains of step (2); Described catalyst is mixture or the cuprous salt of cupric and sodium ascorbate, the molar dose of described catalyst is respectively the molar dose of alkynyl on described silica gel: cupric 1 ~ 10% mixes with sodium ascorbate 2 ~ 40%, or is cuprous salt 1%-10%;
3. react 72 ~ 120 hours under 10 ~ 80 DEG C of conditions, then filter with sand core funnel, wash with the EDETATE SODIUM aqueous solution, water, methyl alcohol and acetone that water, weight concentration are 2 ~ 20% successively, obtain solids;
4. the solids that step (2) 3. obtains is placed in vacuum drying chamber under 40 ~ 80 DEG C of conditions dry 6 ~ 12 hours, obtains target product---dibasic aminoacid hydrophilic chromatographic stationary phase;
The reaction equation of described step (2) is:
，
In formula, the structure of R is:
。
Optionally, the organic solvent that step (1) 1. adopts is DMF.
Further, the structure of silane coupler that step (1) 1. adopts is:
，
Its reaction equation is:
。
Optionally, the full multi-hole blangel orbicule that 2. the used micro-spherical silica gel of step (1) is particle diameter and uniform pore diameter, its particle diameter is 5 ~ 40 μm, and aperture is 60 ~ 300
.
Further, the structure of the dibasic aminoacid (nitrine dibasic aminoacid) that what step (2) 2. added be modified with azido group is:
；
The synthesis step of described nitrine dibasic aminoacid is:
。
Good effect of the present invention is:
(1) dibasic aminoacid hydrophilic chromatographic stationary phase of the present invention adopts dibasic aminoacid as polar functional group, and its character is very stable, and its surface texture can not change because of the change of pH, is also not easy to react with solute molecule; Dibasic aminoacid hydrophilic chromatographic stationary phase of the present invention efficiently can be separated strong polar compound under hydrophilic interaction liquid chromatogram pattern as polar stationary phase.
(2) preparation method of dibasic aminoacid hydrophilic chromatographic stationary phase of the present invention, adopts " click chemistry " as bonding reaction method, can realize the immobilized of high selectivity and high conversion in a mild condition; Its preparation process is simple, and each intermediate structure is clear and definite, can effectively prepare described hydrophilic chromatographic stationary phase.
(3) the present invention has positive role to exploitation hydrophilic chromatographic stationary phase, has very important significance to the commercialization tool of hydrophilic chromatographic stationary phase.Have broad application prospects in some antibiotic such as cephalo and Pei Nan of separation.
Accompanying drawing explanation
Fig. 1 is the FB(flow block) of the preparation method of dibasic aminoacid hydrophilic chromatographic stationary phase of the present invention.
Fig. 2 is the chromatogram that the dibasic aminoacid hydrophilic chromatographic stationary phase dress post of embodiment 1 preparation is separated for nucleosides.
Fig. 3 is that dibasic aminoacid hydrophilic chromatographic stationary phase prepared by embodiment 1 fills the chromatogram of post for Small molecular acid separation.
Fig. 4 is the chromatogram that the dibasic aminoacid hydrophilic chromatographic stationary phase dress post of embodiment 1 preparation is separated for Small molecular alkali.
Fig. 5 is the chromatogram that the dibasic aminoacid hydrophilic chromatographic stationary phase dress post of embodiment 1 preparation is separated for cephalo.
Fig. 6 is that dibasic aminoacid hydrophilic chromatographic stationary phase prepared by embodiment 1 fills post for training the chromatogram of south separation.
Detailed description of the invention
The specific embodiment of the present invention is provided below in conjunction with accompanying drawing.3 embodiments are provided.But it may be noted that enforcement of the present invention is not limited to following embodiment.
embodiment 1
A preparation method for dibasic aminoacid hydrophilic chromatographic stationary phase, uses " click chemistry " to be bonding method bonding dibasic aminoacid molecule, comprises the following steps (see Fig. 1):
(1) Silica Surface introduces terminal acetylene
1. in 500ml there-necked flask, add anhydrous DMF 100ml, 3-isocyanatopropyl triethoxysilane 35mmol(be about 8.66m1), propargylamine 42mmol(is about 2.88mL), react 12 hours at 85 DEG C.
2. add that particle diameter is 5 ~ 40 μm, aperture is 60 ~ 300
, full multi-hole blangel orbicule (note: the approved product the being purchase) 10g of particle diameter and uniform pore diameter.
3. at 110 DEG C, continue reaction 31 hours, then filter with sand core funnel, then wash with carrene 300mL, methyl alcohol 500mL and acetone 250mL successively, obtain solids.
4. the solids that step (1) 3. obtains is placed in vacuum drying chamber under 60 DEG C of conditions dry 7 hours, obtains terminal acetylene silica gel.
(2) " click chemistry " is utilized to prepare dibasic aminoacid hydrophilic chromatographic stationary phase
1. the terminal acetylene silica gel that step (1) 4. obtains being added volume ratio is that in the water of 5 ︰ 1 and the mixed solvent of methyl alcohol, needed for every gram of terminal acetylene silica gel, the amount of water and methanol mixed solvent is 40mL, obtains alkynyl on silica gel
.
2. the dibasic aminoacid and catalyst cupric and sodium ascorbate mixture that are modified with azido group is added, or cuprous salt catalyst.
the preparation method of the nitrine dibasic aminoacid added is:
---in 250mL single port bottle, add iminodiacetic acid 5g, methyl alcohol 100mL, dropwise add thionyl chloride (2.2equiv) under ice-water bath, drip off rear stirred at ambient temperature 3 hours, revolve and steam except desolventizing and thionyl chloride, obtain 7.4g iminodiacetic acid (salt) dimethyl phthalate hydrochloride 2, yield is 100%.
---in 250mL single port bottle, add iminodiacetic acid (salt) dimethyl phthalate hydrochloride 5g, p-nitrobenzyl chloride (1equiv) 4.4g, potash (2.5equiv) 8.8g, DMSO 90mL, stir 8 hours at 50 DEG C; Cooled by reactant liquor, add water 50mL, EA extracts (30mL × 3), and organic phase merges, water washing (30mL
×2), revolve steamed post (PE:EA=8:1) and obtain 5.4g faint yellow solid 3, yield is 72%.
---add nitro amino carbomethoxyphenyl 5g, methyl alcohol 50mL and Raney Ni 0.5g in 100mL single port bottle, under an atmosphere of hydrogen, stirred at ambient temperature 10 hours, TLC detection reaction is complete, suction filtration, revolves steaming, and obtain 4.2g oily compound thing 4, yield is 93%.
---in 250mL single hole bottle, add 8.4g oily compound thing 4, cupric sulfate pentahydrate (0.01equiv) 0.08g, potash (3equiv) 13.1g and methyl alcohol 90mL, stirring ice bath is lowered the temperature, add nitrine sulphonylimidazole hydrochloride (1.5equiv) 9.9g under ice-water bath in batches, add rear stirred at ambient temperature 10 hours, TLC detection reaction, suction filtration revolves steamed post (PE:EA=6:1) and obtains 6.5g brown oil compound 5, yield 71%.
---in 250mL single port bottle, add 6.5g brown oil compound 5, NaOH (4.2equiv) 3.7g and methyl alcohol 50mL, stirred at ambient temperature 5 hours, obtains nitrine dibasic aminoacid 6.TLC detection reaction is complete, and revolve a small amount of reactant liquor of steaming and detect, all the other are directly acidified to pH6 ~ 7 with glacial acetic acid.
The molar dose that described nitrine dibasic aminoacid adds is 2 times of the molar dose of alkynyl on the silica gel that 1. obtains of step (2).
Get the water-soluble solution of sodium ascorbate 0.44g 5mL, get cupric sulfate pentahydrate 0.28g and also use the water-soluble solution of 5mL; Cupric sulfate pentahydrate solution is dropped in sodium ascorbate solution, then mixed solution is dropped in above-mentioned nitrine dibasic aminoacid system, under slowly stirring, add alkynyl on silica gel that 3.4g step (2) 1. obtains.
3. stirring reaction 72 hours under 40 DEG C of conditions, then filter with sand core funnel, wash with methyl alcohol 300mL, 10% EDTA solution 500mL, water 300mL, methyl alcohol 200mL, THF 200mL, acetonitrile 200mL and the carrene 200mL of 50 DEG C successively again, obtain solids.
4. the solids that step (2) 3. obtains is placed in vacuum drying chamber under 50 DEG C of conditions dry 10 hours, obtains target product---dibasic aminoacid hydrophilic chromatographic stationary phase, its structure is:
。
chromatographic evaluation is carried out to dibasic aminoacid hydrophilic chromatographic stationary phase dress post prepared by embodiment 1
Dibasic aminoacid hydrophilic chromatographic stationary phase embodiment 1 prepared fills in the stainless steel HPLC chromatographic column of 4.6mm × 150mm, the reservation being used for obtained chromatographic column under hydrophilic chromatographic pattern testing strong polar compound be separated biased sample.
(1) for the separation of nucleosides
Using acetonitrile/water as mobile phase, the volume ratio of acetonitrile and water is 80 ︰ 20, flow velocity 1mL/min, column temperature is 35 DEG C, determined wavelength is 254nm, and its chromatogram is see Fig. 2 (in figure: 1 be sulphadiazine, 2 be uracil, 3 be uridine, 4 be adenosine, 5 be inosine, 6 be adenine, 7 for guanosine).
Separating resulting shows: in the chromatographic column obtained by dibasic aminoacid hydrophilic chromatographic stationary phase of the present invention, the strong polar compound uracil that the reverse-phase chromatographic column dead time flows out obtains good reservation, and, other nucleosides have also been obtained good baseline separation, show typical hydrophilic interaction pattern feature and good separation selectivity.
(2) for the separation of Small molecular acid
Using acetonitrile/water as mobile phase, the volume ratio of acetonitrile and water is 95 ︰ 5, flow velocity 1mL/min, column temperature is 35 DEG C, determined wavelength is 254nm, and its chromatogram is see Fig. 3 (in figure: 1 be sorbic acid, 2 be p-aminobenzoic acid, 3 be P-hydroxybenzoic acid, 4 for PCA).
Separating resulting shows: in the chromatographic column obtained by dibasic aminoacid hydrophilic chromatographic stationary phase of the present invention, and several Small molecular acid obtains good baseline separation, shows typical hydrophilic interaction pattern feature and good separation selectivity.
(3) for the separation of Small molecular alkali
Using acetonitrile/water as mobile phase, the volume ratio of acetonitrile and water is 95 ︰ 5, flow velocity 1mL/min, and column temperature is 35 DEG C, determined wavelength is 254nm, and its chromatogram is see Fig. 4 (in figure: 1 be caffeine, 2 be theobromine, 3 be theophylline, 4 diprophyllines, 5 are for Quercetin).
Separating resulting shows: in the chromatographic column obtained by dibasic aminoacid hydrophilic chromatographic stationary phase of the present invention, and several Small molecular alkali obtains good baseline separation, shows typical hydrophilic interaction pattern feature and good separation selectivity.
(4) for the separation of cephalo
Using acetonitrile/10mM ammonium formate as mobile phase, the volume ratio of acetonitrile and ammonium formate is 80 ︰ 20, flow velocity 1mL/min, column temperature is 35 DEG C, determined wavelength is 254nm, and its chromatogram is see Fig. 5 (in figure: 1 be Cefamandole Nafate, 2 be cefobutazine sodium, 3 be Cefodizime Sodium, 4 be cephalo diltiazem, 5 be cefotiam hydrochloride, 6 be cefotaxime sodium, 7 be cefepime Hydrochloride, 8 for Cefradine).
Separating resulting shows: in the chromatographic column obtained by dibasic aminoacid hydrophilic chromatographic stationary phase of the present invention, and several cephalo obtains good separation, shows typical hydrophilic interaction pattern feature and good separation selectivity.
(5) for training the separation in south
Using acetonitrile/10mM ammonium formate (pH4.5) as mobile phase, the volume ratio of acetonitrile and ammonium formate is 75 ︰ 25, flow velocity 1mL/min, and column temperature is 35 DEG C, determined wavelength is 254nm, and its chromatogram is see Fig. 6 (in figure: 1 be Biapenem, 2 be Meropenem, 3 for donipenem).
Separating resulting shows: in the chromatographic column obtained by dibasic aminoacid hydrophilic chromatographic stationary phase of the present invention, and several training south obtains good baseline separation, shows typical hydrophilic interaction pattern feature and good separation selectivity.
(6) evaluation result
Through being separated with Pei Nan's nucleosides, Small molecular acid, Small molecular alkali, cephalo, the chromatographic column adopting dibasic aminoacid hydrophilic chromatographic stationary phase of the present invention to obtain shows typical hydrophilic interaction pattern feature and good separation selectivity.
embodiment 2
A preparation method for dibasic aminoacid hydrophilic chromatographic stationary phase, uses " click chemistry " to be bonding method bonding dibasic aminoacid molecule, comprises the following steps:
(1) Silica Surface introduces terminal acetylene
1. in 500ml there-necked flask, add anhydrous DMF 100ml, 3-isocyanatopropyl triethoxysilane 35mmol(be about 8.66m1), propargylamine 70mmol(is about 3.63mL), react 36 hours at 60 DEG C.
2. (with embodiment 1).
3. at 80 DEG C, continue reaction 48 hours, then filter with sand core funnel, then wash with carrene 300mL, methyl alcohol 500mL and acetone 250mL successively, obtain solids.
4. the solids that step (1) 3. obtains is placed in vacuum drying chamber under 40 DEG C of conditions dry 12 hours, obtains terminal acetylene silica gel.
(2) " click chemistry " is utilized to prepare dibasic aminoacid hydrophilic chromatographic stationary phase
1. the terminal acetylene silica gel that step (1) 4. obtains being added volume ratio is that in the water of 8:1 and the mixed solvent of methyl alcohol, needed for every gram of terminal acetylene silica gel, the amount of water and methanol mixed solvent is 40mL, obtains terminal acetylene silica gel.
2. the dibasic aminoacid and catalyst cupric and sodium ascorbate that are modified with azido group is added.
The molar dose that nitrine dibasic aminoacid adds is 1 times of the molar dose of alkynyl on the silica gel that 1. obtains of step (2).
Get the water-soluble solution of sodium ascorbate (0.1equiv) 0.44g 5mL, get cupric sulfate pentahydrate 0.28g and also use the water-soluble solution of 5mL; Cupric sulfate pentahydrate solution is dropped in sodium ascorbate solution, then mixed solution is dropped to above-mentioned nitrine dibasic aminoacid, under slowly stirring, add alkynyl on silica gel that 6.8g step (2) 1. obtains.
3. stirring reaction 120 hours under 10 DEG C of conditions, then filter with sand core funnel, wash with methyl alcohol 300mL, 2% EDTA solution 500mL, water 300mL, methyl alcohol 200mL, THF200mL, the acetonitrile 200mL and carrene 200mL of 50 DEG C successively again, obtain solids.
4. the solids that step (2) 3. obtains is placed in vacuum drying chamber under 40 DEG C of conditions dry 6 hours, obtains target product---dibasic aminoacid hydrophilic chromatographic stationary phase.
embodiment 3
A preparation method for dibasic aminoacid hydrophilic chromatographic stationary phase, uses " click chemistry " to be bonding method bonding dibasic aminoacid molecule, comprises the following steps:
(1) Silica Surface introduces terminal acetylene
1. in 500ml there-necked flask, add anhydrous DMF 100ml, 3-isocyanatopropyl triethoxysilane 35mmol(be about 8.66m1), propargylamine 42mmol(is about 2.88mL), react 6 hours at 150 DEG C.
2. (with embodiment 1).
3. at 120 DEG C, continue reaction 12 hours, then filter with sand core funnel, wash with carrene 300mL, methyl alcohol 500mL and acetone 250mL successively, obtain solids.
4. the solids that step (1) 3. obtains is placed in vacuum drying chamber under 80 DEG C of conditions dry 6 hours, obtains terminal acetylene silica gel.
(2) " click chemistry " is utilized to prepare dibasic aminoacid hydrophilic chromatographic stationary phase
1. the terminal acetylene silica gel that step (1) 4. obtains being added volume ratio is that in the water of 10:1 and the mixed solvent of methyl alcohol, needed for every gram of terminal acetylene silica gel, the amount of water and methanol mixed solvent is 50mL, obtains terminal acetylene silica gel.
2. the dibasic aminoacid and catalyst cupric and sodium ascorbate that are modified with azido group is added.
The molar dose that nitrine dibasic aminoacid adds is 15 times of the molar dose of alkynyl on the silica gel that 1. obtains of step (2).
Get the water-soluble solution of sodium ascorbate (0.1equiv) 0.44g 5mL, get cupric sulfate pentahydrate 0.28g and also use the water-soluble solution of 5mL; Cupric sulfate pentahydrate solution is dropped in sodium ascorbate solution, then mixed solution is dropped in above-mentioned nitrine dibasic aminoacid system, under slowly stirring, add alkynyl on silica gel that 0.5g step (2) 1. obtains.
3. stirring reaction 120 hours under 80 DEG C of conditions, then filter with sand core funnel, wash with methyl alcohol 300mL, 20% EDTA solution 500mL, water 300mL, methyl alcohol 200mL, THF200mL, the acetonitrile 200mL and carrene 200mL of 50 DEG C successively again, obtain solids.
4. the solids that step (2) 3. obtains is placed in vacuum drying chamber under 80 DEG C of conditions dry 12 hours, obtains target product---dibasic aminoacid hydrophilic chromatographic stationary phase.
Claims (6)
1. a dibasic aminoacid hydrophilic chromatographic stationary phase, is characterized in that, its structure is:
。
2. a preparation method for dibasic aminoacid hydrophilic chromatographic stationary phase as claimed in claim 1, uses " click chemistry " to be bonding method bonding dibasic aminoacid molecule, comprises the following steps:
(1) Silica Surface introduces terminal acetylene
1. add silane coupler, propargylamine in organic solvent, the mol ratio of silane coupler and propargylamine is 1 ~ 5:1.2 ~ 5, and needed for every gram of silica gel, the amount of organic solvent is 5 ~ 30mL, reacts 6 ~ 36 hours under 60 ~ 150 DEG C of conditions;
2. add the micro-spherical silica gel of activation, the silane coupler needed for every gram of micro-spherical silica gel is 1 ~ 10mmol;
3. under 80 ~ 120 DEG C of conditions, continue reaction 12 ~ 48 hours, then filter with sand core funnel, more successively with the washing of carrene, methyl alcohol and acetone, obtain solids;
4. the solids that step (1) 3. obtains is placed in vacuum drying chamber under 40 ~ 80 DEG C of conditions dry 6 ~ 12 hours, obtains terminal acetylene silica gel;
It is characterized in that:
(2) by " click chemistry " bonding dibasic aminoacid hydrophilic chromatographic stationary phase
1. the terminal acetylene silica gel that step (1) 4. obtains being added volume ratio is that in the water of 2 ~ 10 ︰ 1 and the mixed solvent of methyl alcohol, needed for every gram of alkynyl silica gel, the amount of water and methanol mixed solvent is 20 ~ 50mL, obtains alkynyl on silica gel;
2. the dibasic aminoacid and catalyst that are modified with azido group is added: the molar dose that the dibasic aminoacid being modified with azido group adds is 1 ~ 15 times of the molar dose of alkynyl on the silica gel that 1. obtains of step (2); Described catalyst is mixture or the cuprous salt of cupric and sodium ascorbate, the molar dose of described catalyst is respectively the molar dose of alkynyl on described silica gel: cupric 1 ~ 10% mixes with sodium ascorbate 2 ~ 40%, or is cuprous salt 1%-10%;
3. react 72 ~ 120 hours under 10 ~ 80 DEG C of conditions, then filter with sand core funnel, wash with the EDETATE SODIUM aqueous solution, water, methyl alcohol and acetone that water, weight concentration are 2 ~ 20% successively, obtain solids;
4. the solids that step (2) 3. obtains is placed in vacuum drying chamber under 40 ~ 80 DEG C of conditions dry 6 ~ 12 hours, obtains target product---dibasic aminoacid hydrophilic chromatographic stationary phase;
The reaction equation of described step (2) is:
，
In formula, the structure of R is:
。
3. preparation method according to claim 2, is characterized in that, the organic solvent that step (1) 1. adopts is DMF.
4. preparation method according to claim 2, is characterized in that, the structure of the silane coupler that step (1) 1. adopts is:
，
Its reaction equation is:
。
5. preparation method according to claim 2, is characterized in that, the full multi-hole blangel orbicule that 2. the used micro-spherical silica gel of step (1) is particle diameter and uniform pore diameter, its particle diameter is 5 ~ 40 μm, and aperture is 60 ~ 300
.
6. preparation method according to claim 2, is characterized in that, the structure being modified with the dibasic aminoacid of azido group that step (2) 2. adds is:
；
The synthesis step of nitrine dibasic aminoacid is:
。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410471342.1A CN104324707A (en) | 2014-09-16 | 2014-09-16 | Amino diacid hydrophilic chromatographic stationary phase and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410471342.1A CN104324707A (en) | 2014-09-16 | 2014-09-16 | Amino diacid hydrophilic chromatographic stationary phase and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104324707A true CN104324707A (en) | 2015-02-04 |
Family
ID=52399585
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410471342.1A Pending CN104324707A (en) | 2014-09-16 | 2014-09-16 | Amino diacid hydrophilic chromatographic stationary phase and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104324707A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104826618A (en) * | 2015-04-20 | 2015-08-12 | 华东理工大学 | Aminodiol hydrophilic chromatography stationary phase and preparation method thereof |
| CN110907548A (en) * | 2019-11-21 | 2020-03-24 | 上海市食品药品检验所 | Method for detecting biapenem and/or related substances |
| CN111408359A (en) * | 2020-01-11 | 2020-07-14 | 华东理工大学 | Polymer type amphoteric hydrophilic interaction chromatographic stationary phase, preparation method thereof and hydrophilic interaction chromatographic column |
| CN113441122A (en) * | 2020-03-24 | 2021-09-28 | 中国科学院大连化学物理研究所 | Chiral stationary phase of spiral polymer functionalized silica gel microsphere, preparation and application |
| CN115245816A (en) * | 2022-07-08 | 2022-10-28 | 河北工业大学 | Amino nicotinic acid mixed-mode chromatographic stationary phase and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101274270A (en) * | 2007-03-30 | 2008-10-01 | 中国科学院大连化学物理研究所 | Method for preparing bonding type cyclodextrin stationary phase with click chemistry reaction |
| CN101690887A (en) * | 2009-10-16 | 2010-04-07 | 华东理工大学 | Oligopeptide phase reversal chromatography stationary phase and preparation method thereof |
| CN101757897A (en) * | 2009-10-16 | 2010-06-30 | 华东理工大学 | Chitosan oligosaccharide hydrophilic interaction chromatography stationary phase and preparation method thereof |
| US20130324700A1 (en) * | 2011-01-28 | 2013-12-05 | Dailian Institute of Chemical Physics, Chinese Aca of Sciences | Zwitterionic stationary phase for hydrophilic interaction liquid chromatography and preparation method thereof |
-
2014
- 2014-09-16 CN CN201410471342.1A patent/CN104324707A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101274270A (en) * | 2007-03-30 | 2008-10-01 | 中国科学院大连化学物理研究所 | Method for preparing bonding type cyclodextrin stationary phase with click chemistry reaction |
| CN101690887A (en) * | 2009-10-16 | 2010-04-07 | 华东理工大学 | Oligopeptide phase reversal chromatography stationary phase and preparation method thereof |
| CN101757897A (en) * | 2009-10-16 | 2010-06-30 | 华东理工大学 | Chitosan oligosaccharide hydrophilic interaction chromatography stationary phase and preparation method thereof |
| US20130324700A1 (en) * | 2011-01-28 | 2013-12-05 | Dailian Institute of Chemical Physics, Chinese Aca of Sciences | Zwitterionic stationary phase for hydrophilic interaction liquid chromatography and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| CHANGHU CHU ET AL.: "Application of click chemistry on preparation of separation materials for liquid chromatography", 《CHEM. SOC. REV.》 * |
| WEI YIN ET AL.: "Click N-benzyl iminodiacetic acid: Novel silica-based tridentate zwitterionic stationary phase for hydrophilic interaction liquid chromatography", 《TALANTA》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104826618A (en) * | 2015-04-20 | 2015-08-12 | 华东理工大学 | Aminodiol hydrophilic chromatography stationary phase and preparation method thereof |
| CN104826618B (en) * | 2015-04-20 | 2017-05-24 | 华东理工大学 | Aminodiol hydrophilic chromatography stationary phase and preparation method thereof |
| CN110907548A (en) * | 2019-11-21 | 2020-03-24 | 上海市食品药品检验所 | Method for detecting biapenem and/or related substances |
| CN111408359A (en) * | 2020-01-11 | 2020-07-14 | 华东理工大学 | Polymer type amphoteric hydrophilic interaction chromatographic stationary phase, preparation method thereof and hydrophilic interaction chromatographic column |
| CN113441122A (en) * | 2020-03-24 | 2021-09-28 | 中国科学院大连化学物理研究所 | Chiral stationary phase of spiral polymer functionalized silica gel microsphere, preparation and application |
| CN113441122B (en) * | 2020-03-24 | 2022-08-09 | 中国科学院大连化学物理研究所 | Chiral stationary phase of spiral polymer functionalized silica gel microsphere, preparation and application |
| CN115245816A (en) * | 2022-07-08 | 2022-10-28 | 河北工业大学 | Amino nicotinic acid mixed-mode chromatographic stationary phase and preparation method and application thereof |
| CN115245816B (en) * | 2022-07-08 | 2024-01-23 | 河北工业大学 | Amino-nicotinic acid mixed mode chromatographic stationary phase and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104324707A (en) | Amino diacid hydrophilic chromatographic stationary phase and preparation method thereof | |
| Qiao et al. | Development and evaluation of new imidazolium-based zwitterionic stationary phases for hydrophilic interaction chromatography | |
| CN101274270B (en) | Method for preparing bonding type cyclodextrin stationary phase with click chemistry reaction | |
| CN101513577B (en) | Melamine molecular imprinted solid phase extraction cartridge and a preparation method and application thereof | |
| Shen et al. | A novel amide stationary phase for hydrophilic interaction liquid chromatography and ion chromatography | |
| CN101757897A (en) | Chitosan oligosaccharide hydrophilic interaction chromatography stationary phase and preparation method thereof | |
| CN102207495A (en) | Method for determining content of glyphosate in soils by using high performance liquid chromatography method | |
| CN103551118B (en) | Column [5] aromatic bonding silica gel stationary phase as well as preparation method and application thereof | |
| CN104277061B (en) | A kind of boric acid fluorescent molecular probe and its preparation method and application | |
| CN102230921A (en) | Detection method for gamma-aminobutyric acid content in food by using high performance liquid chromatography method | |
| CN103816878A (en) | Imidazole zwitterion core-shell hydrophilic-effect stationary phase as well as preparation and application thereof | |
| CN108114707A (en) | A kind of silica matrix hydrophobic amino acid bonded stationary phase and its preparation and application | |
| CN104826618B (en) | Aminodiol hydrophilic chromatography stationary phase and preparation method thereof | |
| CN107837796B (en) | Bonded chromatographic column stationary phase | |
| CN103193898B (en) | L-Phe derives the synthesis of beta-cyclodextrin bonded silica gel and applies the separation of Alanine enantiomers | |
| CN104785230B (en) | Amphoteric bonded silica gel stationary phase as well as preparation method and application thereof | |
| CN102343258B (en) | Triazolyl bonded cyclodextrin-silica gel chiral stationary phase and preparation method thereof | |
| CN106552610B (en) | A kind of hydrophilic Interaction Chromatography/ion-exchange chromatography spectra unmixing and its preparation and application | |
| CN104028253B (en) | A kind of gold nano grain-polysaccharide bonded silica gel stationary phase and preparation method thereof | |
| CN104569372B (en) | The haptenic preparation method of a kind of concealed malachite green | |
| CN115245816B (en) | Amino-nicotinic acid mixed mode chromatographic stationary phase and preparation method and application thereof | |
| Yan et al. | A sulfonated chitooligosaccharide modified silica material for hydrophilic interaction liquid chromatography and its chromatographic evaluation | |
| CN104497058B (en) | The method that the biomolecule of fluorine loaded by carrier is synthesized using polytetrafluoroethylparticle particle | |
| CN100496705C (en) | Hydrophilic NPLC, filler and separating method | |
| CN105642358B (en) | One kind contains vitamin B12Particle emulsifying agents of structure and its preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150204 |
|
| RJ01 | Rejection of invention patent application after publication |