CN102974329B - Preparation method of sterol-based chromatographic stationary phase - Google Patents
Preparation method of sterol-based chromatographic stationary phase Download PDFInfo
- Publication number
- CN102974329B CN102974329B CN201210505338.3A CN201210505338A CN102974329B CN 102974329 B CN102974329 B CN 102974329B CN 201210505338 A CN201210505338 A CN 201210505338A CN 102974329 B CN102974329 B CN 102974329B
- Authority
- CN
- China
- Prior art keywords
- chromatographic stationary
- isocyanic acid
- sterol
- coupling agent
- acid alkyl
- 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.)
- Active
Links
Landscapes
- Steroid Compounds (AREA)
Abstract
The invention discloses a preparation method of a sterol-based chromatographic stationary phase. The sterol-based chromatographic stationary phase is prepared in a manner that a coupling agent reacts with micro-spherical inorganic substrate filler and a sterol compound respectively; and the coupling agent is alkyl-silane isocyanate with the structural formula as shown in the description, wherein X is methyl or ethyl and m is equal to 3-8. The sterol compound is directly used for reacting, so that the preparation process is simple and the cost is low; and the prepared sterol-based chromatographic stationary phase is a pervasive chromatographic stationary phase with a hydrophobic effect, has very good separation selectivity on the vast majority of compounds and can be widely applied to separation of various samples.
Description
Technical field
The invention belongs to liquid chromatography technology field, be specifically related to a kind of preparation method of sterol base chromatographic stationary phases.
Background technology
High performance liquid chromatography (HPLC) is the topmost part of contemporary chromatographic technique, be with fastest developing speed, the most widely used separate analytical technique of Modern Analytical Chemistry, it plays an increasingly important role in various fields such as chemistry, medicine, food, environmental protection, biochemistry and industry preparations.In highly effective liquid phase chromatographic system, to fix and occupy mutually critical role, the development of high selectivity chromatographic stationary phases is the research frontier of Modern Analytical Chemistry, the exploitation of Novel liquid-phase chromatographic stationary phases has very important significance of scientific research and using value.。
Sterol is that a class condenses by 3 hexane rings and a pentamethylene cyclopentanoperhy drophenanthrene derivative forming, it is the important natural active matter of a class being extensively present in organism, can be divided into the three major types such as animality sterol, plant sterols and mushroom sterol by its raw material sources.Animality sterol is taking cholesterol as main, and plant sterols is mainly sitosterol, stigmasterol and campesterol etc.They are not only and form biomembranous important component part, but also are the precursor substances of synthetic many hormones.
The molecular structure of sterol is not only given its extremely strong hydrophobicity, but also make it have unique space structure and liquid crystal property, Just because of this, chromatographic separation material taking steroid as part has caused vast chromatography worker's very big interest in recent years, develop multiple sterol base chromatographic stationary phases, and be widely used in normal-phase chromatography separation steroid hormone compound, reversed phase chromatography separation hydrophobic compound as condensed-nuclei aromatics compounds, fullerene, naphthalene ethylamine optical isomer etc.Such fixing phase not only has the hydrophobic effect feature similar with ODS bonded stationary phase, and along with the variation of separation temperature presents certain liquid crystal property, fixing phase can not occur under high aqueous phase system condition " caves in and " and causes solute to retain the phenomenon reducing, in addition, the introducing of some electronegative elements (as O, N), makes this fixing because the appearance (as ether, formic acid esters, ester) of some polar group presents certain polarity feature.
The sterols Stationary phase preparation method of report mainly contains two kinds of methods at present: a kind of method is first silica gel to be reacted with triethoxysilane to generate hydrosilation silica gel, then under Karstedt catalyst action, by hydrosilylation, cholesterol unsaturated acid ester is fixed on to Silica Surface (J.J.Pesek, M.T.Matyska, E.J.Williamsen, R.Tam, Chromatographia, 41 (1995) 301-310), such is fixing mutually by the commercialization of MicroSolv company of the U.S.; The operation of another kind method is relatively easy---and the silica gel Direct Bonding by sterol chloro-formate and γ-aminopropylsilane modification forms (B.Buszewski, M.Jezierska, O.G. Barbara.Materials Chemistry and Physics, 72 (2001) 30 – 41).But two kinds of methods all need first sterol to be carried out to modification, have significantly increased cost of material, and: the reaction condition of first method is also comparatively strict, and catalyst is very expensive; The part raw material sterol chloro-formate that second method is used is except the relatively unmodified sterol of cost is higher, and character is also more active, more responsive to air humidity.
Summary of the invention
The object of the present invention is to provide a kind of sterol primary colours spectrum Stationary phase preparation method.This Stationary phase preparation method is simple, with low cost, of many uses.
For achieving the above object, the technical solution used in the present invention is:
A preparation method for sterol base chromatographic stationary phases, described sterol base chromatographic stationary phases is reacted and makes with the inorganic matrix filler of microspheroidal and phytosterin compound respectively by coupling agent, and described coupling agent is the isocyanic acid alkyl silane with following general structure:
Wherein, X is methyl or ethyl, m=3～8.
In such scheme, described coupling agent first reacts with the inorganic matrix filler of microspheroidal, and then reacts with phytosterin compound, and concrete steps are as follows:
1) in the anhydrous toluene solution that is dissolved with isocyanic acid alkyl silane coupling agent, add microspheroidal inorganic matrix filler, under inert atmosphere, add triethylamine as catalyst, back flow reaction 8～24 hours at 100～160 DEG C, through cooling, suction filtration, washing, vacuum drying, obtain isocyanic acid alkyl silane coupling agent bonded silica gel;
2) then, isocyanic acid alkyl silane coupling agent bonded stationary phase is scattered in dry toluene, under inert atmosphere, adds phytosterin compound and pyridine, after stirring and dissolving, at 50～100 DEG C, react 8～24 hours, through cooling, suction filtration, washing, dry, obtain sterol base chromatographic stationary phases.
In such scheme, described coupling agent first reacts with phytosterin compound, and then reacts with the inorganic matrix filler of microspheroidal, and concrete steps are as follows:
1) in the anhydrous toluene solution that is dissolved with phytosterin compound, dropwise add isocyanic acid alkyl silane coupling agent, under inert atmosphere, add pyridine, stirring reaction 8～24 hours at 50～100 DEG C;
2) then, in reaction system, add microspheroidal inorganic matrix material, under inert atmosphere, add triethylamine as catalyst, back flow reaction 8～24 hours, through cooling, suction filtration, washing, vacuum drying, obtains sterol base chromatographic stationary phases at 100～160 DEG C.
In such scheme, described phytosterin compound includes but not limited to cholesterol (cholesterol), sitosterol (sitosterol), stigmasterol (stigmasterol), campesterol (campesterol), mycosterol (fungisterol).
In such scheme, described inorganic matrix filler includes but not limited to silica gel, zirconia, titanium oxide, aluminium oxide.
In such scheme, the consumption of described isocyanic acid alkyl silane is 1～6mmol/ gram of inorganic matrix filler, and the mol ratio of isocyanic acid alkyl silane and phytosterin compound is 1:1～1:8.
Tool of the present invention has the following advantages:
1. preparation process is simple, with low cost.The present invention is directly bonded in phytosterin compound inorganic matrix filling surface using isocyanic acid alkyl silane coupling agent as " spacerarm " and forms fixing phase, both simplify preparation process, avoided again the derivative of the phytosterin compound of the higher and less stable of use cost simultaneously.
2. applied range.Sterol base chromatographic stationary phases provided by the invention is the fixing phase of a kind of general type hydrophobic interaction chromatography, and most compounds are all had to good separation selectivity, can be widely used in all kinds of sample separation.
Brief description of the drawings
Fig. 1 is benzene and four kinds of homologue (a: benzene thereof; B: toluene; C: ethylbenzene; D: propyl benzene; E: butylbenzene) chromatographic fractionation figure
Fig. 2 is three kinds of Hydroxyphenol Isomers (a: hydroquinones; B: resorcinol; C: catechol) chromatographic fractionation figure
Detailed description of the invention
Below in conjunction with the drawings and specific embodiments, the present invention will be further described.Specific embodiment only limits to illustrate the present invention, but not limitation of the invention.
Embodiment 1
In the anhydrous toluene solution (150mL) that is dissolved with 5mmol3-isocyanic acid propyl trimethoxy silicane, add at 120 DEG C 5 grams of the silica gel microballs of dry 6 hours, in nitrogen atmosphere taking triethylamine as catalyst, at 100 DEG C, react 8 hours, cooling, suction filtration, washing, vacuum drying makes isocyanic acid bonded silica gel, then by isocyanic acid bonded silica gel and cholesterol (5mmol) in dry toluene (150mL), in nitrogen atmosphere taking pyridine as catalyst, at 50 DEG C, react 24 hours, cooling, suction filtration, washing, gained solid is dried 24 hours under 40 DEG C of conditions in vacuum drying chamber, obtain the fixing phase of cholesteryl silica gel chromatograph.This fixing phase structure is:
Embodiment 2
In dry toluene (150mL) solution that is dissolved with cholesterol (40mmol), dropwise add 6-isocyanic acid hexyl triethoxysilane (5mmol), in argon atmosphere taking pyridine as catalyst, stirring reaction 12 hours at 100 DEG C, then in reaction system, add through 5 grams of 160 DEG C of silica gel microballs of dry 10 hours, taking triethylamine as catalyst, at 100 DEG C, react 8 hours, cooling, suction filtration, washing, gained solid is dried 12 hours under 60 DEG C of conditions in vacuum drying chamber, obtains the fixing phase of cholesteryl silica gel chromatograph.This fixing phase structure is:
Embodiment 3
In the anhydrous toluene solution (150mL) that is dissolved with 30mmol3-isocyanic acid propyl trimethoxy silicane, add at 120 DEG C 5 grams of the zirconium oxide microballoons of dry 6 hours, in nitrogen atmosphere taking triethylamine as catalyst, at 100 DEG C, react 8 hours, cooling, suction filtration, washing, vacuum drying makes the fixing phase of isocyanic acid coupling agent bonding zirconia, then itself and campesterol (campesterol) (30mmol) are disperseed, be dissolved in dry toluene (150mL), in nitrogen atmosphere taking pyridine as catalyst, stirring reaction 12 hours at 75 DEG C, through cooling, suction filtration, after carrying out washing treatment, gained solid is dried 12 hours under 60 DEG C of conditions in vacuum drying chamber, obtain rape oil steroid
Embodiment 4
In the anhydrous toluene solution (150mL) that is dissolved with 15mmol3-NCO propyl-triethoxysilicane, add under 160C 5 grams of the titania microspheres of dry 6 hours, in nitrogen atmosphere taking triethylamine as catalyst, at 160 DEG C, react 8 hours, cooling, suction filtration, washing, vacuum drying makes the fixing phase of isocyanic acid coupling agent bonding titanium oxide, then itself and sitosterol (sitosterol) (30mmol) are disperseed, be dissolved in dry toluene (150mL), in nitrogen atmosphere taking pyridine as catalyst, stirring reaction 8 hours at 100 DEG C, through cooling, suction filtration, after carrying out washing treatment, gained solid is dried 24 hours under 40 DEG C of conditions in vacuum drying chamber, obtain sitosterol based titanium oxide chromatographic stationary phases.This fixing phase structure is:
Embodiment 5
Dropwise add 3-isocyanic acid propyl-triethoxysilicane (20mmol) being dissolved with in mycosterol (fungisterol) dry toluene (150mL) solution (20mmol), in nitrogen atmosphere taking pyridine as catalyst, stirring reaction 24 hours at 100 DEG C, then in reaction system, add through 5 grams of 140 DEG C of aluminum oxide micro-spheres of dry 10 hours, taking triethylamine as catalyst, at 150 DEG C, react 18 hours, cooling, suction filtration, washing, gained solid is dried 12 hours under 60 DEG C of conditions in vacuum drying chamber, obtain mycosterol base aluminium oxide chromatographic stationary phases.This fixing phase structure is:
Accompanying drawing provides the cholesteryl silica gel chromatograph of preparation in the embodiment of the present invention 1 fixing phase actual separation design sketch, and wherein Fig. 1 is benzene and four kinds of homologue (a: benzene thereof; B: toluene; C: ethylbenzene; D: propyl benzene; E: butylbenzene) chromatographic fractionation figure, Fig. 2 is three kinds of Hydroxyphenol Isomers (a: hydroquinones; B: resorcinol; C: catechol) chromatographic fractionation figure.From spectrogram, can find out that the fixing relative target separator of cholesteryl silica gel chromatograph has good separating effect.
Claims (5)
1. a preparation method for sterol base chromatographic stationary phases, is characterized in that: described sterol base chromatographic stationary phases is reacted and makes with the inorganic matrix filler of microspheroidal and phytosterin compound respectively by coupling agent;
Described coupling agent is the isocyanic acid alkyl silane with following general structure:
Wherein, X is methyl or ethyl, m=3～8,
Described inorganic matrix filler is the one in silica gel, zirconia, titanium oxide and aluminium oxide;
Concrete steps are:
1) in the anhydrous toluene solution that is dissolved with isocyanic acid alkyl silane coupling agent, add microspheroidal inorganic matrix filler, under inert atmosphere, add triethylamine as catalyst, back flow reaction 8～24 hours at 100～160 DEG C, through cooling, suction filtration, washing, vacuum drying, obtain isocyanic acid alkyl silane coupling agent bonded stationary phase;
2) then, isocyanic acid alkyl silane coupling agent bonded stationary phase is scattered in dry toluene, under inert atmosphere, adds phytosterin compound and pyridine, after stirring and dissolving, at 50～100 DEG C, react 8～24 hours, through cooling, suction filtration, washing, dry, obtain sterol base chromatographic stationary phases.
2. a preparation method for sterol base chromatographic stationary phases, is characterized in that: described sterol base chromatographic stationary phases is reacted and makes with the inorganic matrix filler of microspheroidal and phytosterin compound respectively by coupling agent;
Described coupling agent is the isocyanic acid alkyl silane with following general structure:
Wherein, X is methyl or ethyl, m=3～8,
Described inorganic matrix filler is the one in silica gel, zirconia, titanium oxide and aluminium oxide;
Concrete steps are:
1) in the anhydrous toluene solution that is dissolved with phytosterin compound, dropwise add isocyanic acid alkyl silane coupling agent, under inert atmosphere, add pyridine, stirring reaction 8～24 hours at 50～100 DEG C;
2) then, in reaction system, add microspheroidal inorganic matrix material, under inert atmosphere, add triethylamine as catalyst, back flow reaction 8～24 hours, through cooling, suction filtration, washing, vacuum drying, obtains sterol base chromatographic stationary phases at 100～160 DEG C.
3. the preparation method of sterol base chromatographic stationary phases according to claim 1 and 2, is characterized in that: described phytosterin compound is the one in cholesterol, sitosterol, stigmasterol, campesterol and mycosterol.
4. the preparation method of sterol base chromatographic stationary phases according to claim 1 and 2, it is characterized in that: the consumption of described isocyanic acid alkyl silane is 1～6mmol/ gram of inorganic matrix filler, and the mol ratio of isocyanic acid alkyl silane and phytosterin compound is 1:1～1:8.
5. the preparation method of sterol base chromatographic stationary phases according to claim 3, it is characterized in that: the consumption of described isocyanic acid alkyl silane is 1～6mmol/ gram of inorganic matrix filler, and the mol ratio of isocyanic acid alkyl silane and phytosterin compound is 1:1～1:8.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210505338.3A CN102974329B (en) | 2012-11-30 | 2012-11-30 | Preparation method of sterol-based chromatographic stationary phase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210505338.3A CN102974329B (en) | 2012-11-30 | 2012-11-30 | Preparation method of sterol-based chromatographic stationary phase |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102974329A CN102974329A (en) | 2013-03-20 |
| CN102974329B true CN102974329B (en) | 2014-08-06 |
Family
ID=47848846
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210505338.3A Active CN102974329B (en) | 2012-11-30 | 2012-11-30 | Preparation method of sterol-based chromatographic stationary phase |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102974329B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104841375A (en) * | 2015-05-08 | 2015-08-19 | 西北大学 | Preparation and application of high performance hydrophobic interaction chromatography packing material taking cholesterol as aglucon |
| CN108176386B (en) * | 2018-01-16 | 2020-11-06 | 武汉轻工大学 | Melamine bonded chromatographic stationary phase, preparation method thereof and metal ion modified melamine bonded chromatographic stationary phase |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5374755A (en) * | 1992-07-17 | 1994-12-20 | Millipore Corporation | Liquid chromatography stationary phases with reduced silanol interactions |
| CN1116933C (en) * | 1999-08-04 | 2003-08-06 | 中国科学院化学研究所 | Ester-type efficient liquid-phase chromatographic reverse filler and its preparing process |
| CN101670277A (en) * | 2009-09-23 | 2010-03-17 | 华东理工大学 | Preparation method of liquid chromatographic chiral stationary phase of curcumin bonded silica gel |
| CN102101044B (en) * | 2009-12-18 | 2013-01-09 | 中国科学院大连化学物理研究所 | Method for preparing oligomeric ethylene glycol stationary phase |
-
2012
- 2012-11-30 CN CN201210505338.3A patent/CN102974329B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN102974329A (en) | 2013-03-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102172519B (en) | Silica gel bonded cellulose derivative chromatographic filler and preparation method and use thereof | |
| CN103041792B (en) | Carbamic acid ester type liquid phase chromatogram stationary phase and preparation method thereof | |
| CN101987293B (en) | Chromatographic separation material based on copolymerization on silica gel surface and preparation thereof | |
| CN103301822B (en) | A kind of polar liquid chromatogram filler and preparation method thereof | |
| CN101829551B (en) | Silica gel fixed phase of mixed model as well as preparation method and application thereof | |
| CN102974329B (en) | Preparation method of sterol-based chromatographic stationary phase | |
| CN109126749A (en) | A kind of polyhedral oligomeric silsesquioxane is the hud typed chromatographic stationary phases and the preparation method and application thereof of shell | |
| CN107126943A (en) | Phenyl boric acid modified silica-gel function chromatograph packing material, preparation method and application | |
| CN101417227B (en) | Silica gel bonded double-chirality active center chromatogram filler and use thereof | |
| CN103357390A (en) | Multi-layer structure bonded silica gel liquid chromatography packing and synthesis method thereof | |
| CN104826616A (en) | HPLC (high-performance liquid chromatography) chromatographic column packing modification method | |
| CN107478731A (en) | The pre-treating method of parabens preservative in a kind of detection cosmetics | |
| CN102101046A (en) | Carbamido chromatograph stationary phase and preparation method thereof | |
| CN106824140B (en) | A method of chromatographic stationary phases are prepared by template of divinylsulfone | |
| JPH10510633A (en) | Method for modifying a silanol-containing or silanol-containing stationary carrier phase with calixarene for chromatography | |
| CN104785230A (en) | Amphoteric bonded silica gel stationary phase as well as preparation method and application thereof | |
| CN100386142C (en) | Method of fust preparing linkage type polysaccharide chiral fixed phase | |
| CN105983392B (en) | Stationary Phase for HPLC and its preparation method and application based on three cation liquid | |
| CN108079981A (en) | A kind of tetrazole/hydrophobic group bonded stationary phase and preparation method thereof | |
| CN102335598A (en) | Preparation method of phenothiazine bonded silica gel | |
| CN102233263A (en) | Amphoteric imidazoline bonded silica gel stationary phase and preparation method thereof | |
| CN106008982A (en) | Preparation method of boric acid silica gel pretreatment material | |
| CN103203225B (en) | Weak cation exchange/hydrophobic bifunctional hybrid-mode chromatographic stationary phase as well as preparation method and application thereof | |
| CN101628225B (en) | 1-vinyl iminazole ionic liquid chromatography packing as well as preparation method and use method thereof | |
| CN108114706A (en) | A kind of silica matrix taurine bonded stationary phase and its preparation 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 | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |