CN106732475A - A kind of HPLC chiral stationary phases and its preparation method and application - Google Patents
A kind of HPLC chiral stationary phases and its preparation method and application Download PDFInfo
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- 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
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- 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
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- C07C213/00—Preparation of compounds containing amino and hydroxy, amino and etherified hydroxy or amino and esterified hydroxy groups bound to the same carbon skeleton
- C07C213/10—Separation; Purification; Stabilisation; Use of additives
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- C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
- C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
- C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
- C07D213/36—Radicals substituted by singly-bound nitrogen atoms
- C07D213/38—Radicals substituted by singly-bound nitrogen atoms having only hydrogen or hydrocarbon radicals attached to the substituent nitrogen atom
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- C07D279/10—1,4-Thiazines; Hydrogenated 1,4-thiazines
- C07D279/14—1,4-Thiazines; Hydrogenated 1,4-thiazines condensed with carbocyclic rings or ring systems
- C07D279/18—[b, e]-condensed with two six-membered rings
- C07D279/20—[b, e]-condensed with two six-membered rings with hydrogen atoms directly attached to the ring nitrogen atom
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- C07D279/10—1,4-Thiazines; Hydrogenated 1,4-thiazines
- C07D279/14—1,4-Thiazines; Hydrogenated 1,4-thiazines condensed with carbocyclic rings or ring systems
- C07D279/18—[b, e]-condensed with two six-membered rings
- C07D279/22—[b, e]-condensed with two six-membered rings with carbon atoms directly attached to the ring nitrogen atom
- C07D279/24—[b, e]-condensed with two six-membered rings with carbon atoms directly attached to the ring nitrogen atom with hydrocarbon radicals, substituted by amino radicals, attached to the ring nitrogen atom
- C07D279/26—[b, e]-condensed with two six-membered rings with carbon atoms directly attached to the ring nitrogen atom with hydrocarbon radicals, substituted by amino radicals, attached to the ring nitrogen atom without other substituents attached to the ring system
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- B01J2220/00—Aspects relating to sorbent materials
- B01J2220/50—Aspects relating to the use of sorbent or filter aid materials
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Abstract
The present invention relates to a kind of HPLC chiral stationary phases and its preparation method and application, with the good silicon dioxide microsphere of monodispersity as raw material, photosensitive high molecular diazo resin is surface modifier, beta cyclodextrin/polyacrylic acid composite is connected to silicon dioxide microsphere surface, then by ultraviolet light, by the converting hydrogen bonds between the hydrogen bond between diazo resin and silicone hydroxyl, diazo resin and beta cyclodextrin/polyacrylic acid composite be covalent bond so that beta cyclodextrin stabilization be connected to microsphere surface.The photosensitive high molecular diazo resin that the present invention is used, compared with conventional silanes coupling agent, with nonhazardous, it is easily-synthesized, the features such as environment friendly, obtained modified silicon ball chiral stationary phase realizes baseline separation in high performance liquid chromatography to various chiral drugs correspondence body, shows excellent chiral resolution ability, can be applied to various chiral chromatogram column packing fields.
Description
Technical field
The invention belongs to technical field of chromatography separation, and in particular to one kind is existed using photosensitive high molecular polymer diazo resin
HPLC chiral stationary phases that silicon dioxide microsphere surface modification cyclodextrin is modified and its preparation method and application.
Background technology
With the progress and development of society, people gradually recognize chiral drug and live closely bound up, and chiral drug
Also risen year by year in the share of medical market.With sharply increasing for the chiral pharmaceutical requirements of people, quickly chiral point is set up
From method, all have to pharmacokinetic and chiral drug control of product quality and be of great significance.At present, chromatography
The most useful method of optical resolution is turned into.These methods include gas chromatography, thin-layered chromatography, supercritical fluid chromatography
Method, Capillary Electrophoresis, adverse current chromatogram, centrifugal partition chromatograph method, high performance liquid chromatography (HPLC).High performance liquid chromatography by feat of
It is efficiently, high sensitivity, the advantage such as analyze speed is fast, operation strategies are wide be as one, chiral separation field important technology.
Liquid chromatography chiral resolution can be divided into chiral derivatization method, chiral mobile phases and chiral stationary phase according to its principle
Method.Chiral Stationary Phases are the energy for forming temporary transient diastereomer complex with the chiral selector of fixed phase surface based on sample
Measure difference or stability difference and reach chiral separation, be the method without the resolution for being transformed into diastereomer.It is chiral
The advantage of fixing phase Split Method is to prepare conveniently, can be suitably used for the fractionation of classes of compounds.Chiral Stationary Phases be at present most
The Optical Instruments Industry method of advantage.
In chiral chromatogram separation, a kind of preferable chiral stationary phase should possess advantages below：Separating ranges are wide, adapt to many
Plant the Chiral Separation of structure type；Analyze speed is fast, and the rate of departure is high, can the quick, purity of Accurate Determining enantiomer；Separate
With specificity；Separation selectivity and column capacity are higher, with preparative separation ability.In the past few years, substantial amounts of chirality
Fixing phase has obtained exploitation and application, the commodity chiral stationary phase for having synthesized up to hundreds of, but so far, also without one kind
Chiral stationary phase can simultaneously meet the requirement of above-mentioned each side.
Beta cyclodextrin (Cyclodextrins, β CD) is bonded by α-Isosorbide-5-Nitrae-glucosides by a number of glucose unit
The ring molecule for connecing.Molecule is on the whole with the outer hydrophilic characteristic of hydrophobic and ring in ring and stronger chiral recognition
Occupy important status in the separation of chiral drug.Armstrong etc. takes the lead in by alkylation, is acylated and amino first
It is Esterification, new chiral Recognition site is increased, the chiral chromatogram performance of cyclized polyisoprene is greatly improved.And use
Different derivatization reagents is modified the hydroxyl of β CD, can improve the active force between Bonded Phase and chiral molecules, is expanded
Range of application.Cyclodextrin chromatographic stationary phases are usually that β CD or derivatives thereof are bonded on silica matrix, and silica gel first passes through silicon
Alkylators are pre-processed, and conventional silylating reagent has gamma-aminopropyl-triethoxy-silane (KH550), y- glycidyl ether oxygens
Propyl trimethoxy silicane (KH560) and γ-methacryloxypropyl trimethoxy silane (KH570).β CD are by modification
Afterwards, amino, hydroxyl, alkenyl or epoxy functionality are often connected with, can be bonded with silanized silica gel；Also use silylating reagent incites somebody to action
β CD derivatizations, then react with the silicone hydroxyl of silica gel in anhydrous conditions, will β CD be connected on silica matrix.
CN103601823A discloses a kind of preparation method of β open loops dextrin chiral stationary phase, with SiO2Microballoon is raw material,
By gamma-aminopropyl-triethoxy-silane (KH550), trim,ethylchlorosilane, 4,4 '-methyl diphenylene diisocyanate (MDI),
Beta-schardinger dextrin is bonded to SiO2Microsphere surface, obtains a kind of beta-schardinger dextrin chiral stationary phase.CN104841408A discloses one
Cyclodextrin chiral fixing phase and preparation method and application is planted, sulfydryl-vinyl bonding cyclodextrin hand is prepared by click-reaction
Property fixing phase chiral chromatogram filler, processed by modified and silica gel the sulfhydrylation of the selectivity to cyclodextrin 6, using efficiently urging
Agent is by click-reaction and then prepares the cyclodextrin chiral fixing phase that the excellent sulfydryl of chemical stability-vinyl is bonded.With
Upper two methods have used silane coupler when cyclodextrin chiral fixing phase is prepared, and the silane idol commonly used in modifying process
Connection agent often has certain toxicity and to water sensitive, it is necessary to control to use in anhydrous conditions, relatively harsh, and process
It is cumbersome, take more long；In addition, the problems such as these factors can cause environmental pollution, greatly limit its application.
The content of the invention
Therefore, the technical problems to be solved by the invention be existing HPLC chiral stationary phases preparation method complicated and time consumption and
Environment is unfriendly, and then provides a kind of preparation method of new HPLC chiral stationary phases, and the preparation method is using nontoxic photosensitive
, first be grafted to β CD on polyacrylic acid (PAA) chain to increase its water solubility, so by macromolecule diazo resin substituted silane coupling agent
Afterwards by β CD modifications to silica, so as to introduce chiral Recognition group in microsphere surface, reaction condition is gentle, and the time is short, easily
Operation, and modified chromatographic column filler has good chiral separation effect.
In order to solve the above technical problems, the present invention is adopted the following technical scheme that：
A kind of preparation method of HPLC chiral stationary phases, comprises the steps：
S1, silicon dioxide microsphere is dispersed in diazo resin solution, lucifuge is stirred 2-4 hour at room temperature, be centrifuged
Wash microballoon after separation with water；
S2, the product of step S1 is placed in beta-schardinger dextrin/polyacrylic acid composite solution, at room temperature lucifuge stirring 2-4
Hour, wash microballoon after centrifugation with water；
S3, will step S2 microballoon dry after be placed in exposed under UV light treatment 10-20min, obtain can be used as HPLC hands
The improved silica microballoon of property fixing phase.
The preparation method also includes：
S0, silicon dioxide microsphere is evenly dispersed in concentration be 0.08-0.12mol/L hydrochloric acid solution in activated
Treatment 15-45min, washes microballoon to neutrality with water after centrifugation.
The silicon dioxide microsphere is the monodisperse particles of 1-3 μm of average grain diameter.
Diazo resin solution is the diazo resin aqueous solution of 8-12mg/mL, β-ring in the step S2 in the step S1
Dextrin/polyacrylic acid composite solution is the beta-schardinger dextrin/polyacrylic acid composite aqueous solution of 8-12mg/mL.The step S0,
It is microballoon to be washed with deionized 2-4 times to wash microballoon in S1, S2 with water.It is to be put into microballoon that microballoon is dried in the step S3
Carried out in vacuum tank drying at room temperature 12-14 hours.
The HPLC chiral stationary phases that a kind of methods described is prepared.
Application of the described HPLC chiral stationary phases in separating chiral material, comprises the steps：
During HPLC chiral stationary phase packing column machines are filled in into stainless steel chromatographic column, column temperature be 20~30 DEG C, flow velocity be
Under the conditions of 0.2mL/min, chiral material sample carries out high performance liquid chromatography separation.
Described chiral material sample is metoprolol tartrate, fenazil, Dioxopromethazine or chlorpheniramine.
Above-mentioned technical proposal of the invention has advantages below compared to existing technology：
(1) the HPLC chiral stationary phase preparation methods that the present invention is provided are to be met β CD using photosensitive high molecular diazo resin
To SiO2Microsphere surface.After illumination, hydrogen bond action, diazo resin and cyclodextrin surface hydroxyl between diazo resin and silicone hydroxyl
Between hydrogen bond action be converted into covalent bond effect so that β CD stabilization be connected to microsphere surface, obtain β CD-PAA@SiO2It is micro-
Ball.βCD-PAA@SiO2Microballoon is used as liquid chromatographic packing materials, and chiral medicine has stronger fractionation ability.The letter of this preparation method
Just and efficiency high, modified chromatographic column filler has good separating effect.
(2) starting silica microballoon of the invention carries out activation process using hydrochloric acid, and the purpose of activation is raising two
The silicone hydroxyl content on silicon oxide microsphere surface, stabilizes it uniformly presence, improves the degree of modification of follow-up modification.
(3) method of the present invention is used, process equipment is simple, reproducible, raw materials used to be easy to get, low production cost and effect
Rate is high；Additionally, the photosensitive high molecular diazo resin that the present invention is used, compared with conventional silanes coupling agent, with nonhazardous, Yi He
The features such as into, environment friendly.
(4) monodisperse silica microspheres being modified by the inventive method, as performance liquid chromatographic column filler, not
With baseline separation is realized to various enantiomers of chiral drugs under flowing facies model, have broad application prospects.
Brief description of the drawings
Fig. 1 is the scanning electron microscopic picture of the monodisperse silica microspheres used by embodiment 1；
Fig. 2 is the modified Fourier turn infrared with unmodified silicon dioxide microsphere of the gained of embodiment 1
Curve picture；
Fig. 3 is the separation spectrogram of the high performance liquid chromatography of the gained of application examples 1；
Fig. 4 is the separation spectrogram of the high performance liquid chromatography of the gained of application examples 2；
Fig. 5 is the separation spectrogram of the high performance liquid chromatography of the gained of application examples 3；
Fig. 6 is the separation spectrogram of the high performance liquid chromatography of the gained of application examples 4.
Specific embodiment
In order that the object, technical solutions and advantages of the present invention are clearer, below in conjunction with accompanying drawing to reality of the invention
The mode of applying is described in further detail.The present invention can be embodied in many different forms, and should not be construed as limited to
Embodiment set forth herein.Conversely, there is provided these embodiments so that the disclosure will be thorough and complete, and this will be sent out
Bright design is fully conveyed to those skilled in the art, and the present invention will only be defined by the appended claims.
Stereoscan photograph is measured by JEOLJSM-6390LV type ESEMs.
Fourier turn infrared picture is measured by Nicolet6700 type infrared spectrometers.
High-efficient liquid phase chromatogram is measured by the rainbow SEV P500 type liquid chromatographs of Qingdao seven.
The diazo resin used in embodiment 1-4 has structure shown in formula (1)：
Wherein n is the integer of 3-6.
The beta-schardinger dextrin used in embodiment 1-4/polyacrylic acid composite β CD-PAA have structure shown in formula (2)：
Black bell-shaped thing is beta cyclodextrin in formula (2).
Selected β CD-PAA compounds, its preparation process is as follows：By 0.5g amination β cyclodextrins, 0.2g dicyclohexyl carbon
Diimine, 1.4gPAA adds 40mL1- N-methyl-2-2-pyrrolidone Ns, 60 DEG C of reaction 12h to filter product and wash three with methyl alcohol
All over.
Embodiment 1
The preparation method of the HPLC chiral stationary phases of the present embodiment, comprises the steps：
The activation process of S0, silicon dioxide microsphere：
The monodisperse silica microspheres that 2g average grain diameters are 2 μm are evenly dispersed in the hydrochloric acid that concentration is 0.1mol/L
30min is processed in the aqueous solution, simultaneously silicon dioxide microsphere is washed with deionized to neutrality for centrifugation；
S1, the silicon dioxide microsphere after activation process is modified：
By the silicon dioxide microsphere after activation be placed in concentration be 10mg/mL the diazo resin aqueous solution in, at room temperature lucifuge,
Magnetic agitation 3 hours, microballoon is washed with deionized 3 times after centrifugation；
S2, the microballoon that step S1 is obtained is placed in the β CD-PAA aqueous solution that concentration is 10mg/mL, lucifuge, magnetic at room temperature
Power is stirred 3 hours, and microballoon is washed with deionized 3 times after centrifugation.
S3, the microballoon that step S2 is obtained is put in drying at room temperature 12h in vacuum drying chamber, dried microballoon is placed in purple
Exposure-processed 15min under outer light, obtains can be used as the improved silica microballoon β CD-PAA@SiO of HPLC chiral stationary phases2.This
The diazo resin of embodiment is structure, wherein n=4 shown in formula (1).
Inventor has carried out Morphology analysis using SEM to selected silicon dioxide microsphere raw material, such as
Shown in Fig. 1, it can be seen that microballoon monodispersity is good, uniform particle diameter, the particle diameter of microballoon is about 2 μm；In addition, using Fu
In leaf transformation infra-red sepectrometry analyzed with the infrared spectrogram of unmodified silicon dioxide microsphere modified, such as Fig. 2 institutes
Show, wherein a lines are the infrared analysis spectrogram of silicon dioxide microsphere before modified, b lines are the silica after surface modification β CD-PAA
The infrared analysis spectrogram of microballoon, 1093cm in figure-1Place is Si-O vibration peaks, 1639cm in silica-1And 1384cm-1Place is
C-H vibration absorption peaks on cyclodextrin.
Embodiment 2
The preparation method of the HPLC chiral stationary phases of the present embodiment, comprises the steps：
The activation process of S0, silicon dioxide microsphere：
The monodisperse silica microspheres that 2g average grain diameters are 3 μm are evenly dispersed in the salt that concentration is 0.08mol/L
45min is processed in aqueous acid, simultaneously silicon dioxide microsphere is washed with deionized to neutrality for centrifugation；
S1, the silicon dioxide microsphere after activation process is modified：
By the silicon dioxide microsphere after activation be placed in concentration be 12mg/mL the diazo resin aqueous solution in, at room temperature lucifuge,
Magnetic agitation 2 hours, microballoon is washed with deionized 3 times after centrifugation；
S2, the microballoon that step S1 is obtained is placed in the β CD-PAA aqueous solution that concentration is 12mg/mL, lucifuge, magnetic at room temperature
Power is stirred 2 hours, and microballoon is washed with deionized 3 times after centrifugation.
S3, the microballoon that step S2 is obtained is put in drying at room temperature 12h in vacuum drying chamber, dried microballoon is placed in purple
Exposure-processed 20min under outer light, obtains can be used as the improved silica microballoon β CD-PAA@SiO of HPLC chiral stationary phases2。
The diazo resin of the present embodiment is structure, wherein n=5 shown in formula (1).
Embodiment 3
The preparation method of the HPLC chiral stationary phases of the present embodiment, comprises the steps：
S0, the monodisperse silica microspheres that 2g average grain diameters are 1 μm are evenly dispersed in concentration for 0.12mol/L
15min is processed in aqueous hydrochloric acid solution, silicon dioxide microsphere is simultaneously washed to neutrality by centrifugation；
S1, the silicon dioxide microsphere after activation process is modified：
By the silicon dioxide microsphere after activation be placed in concentration be 8mg/mL the diazo resin aqueous solution in, at room temperature lucifuge,
Magnetic agitation 4 hours, microballoon is washed with deionized 3 times after centrifugation；
S2, the microballoon that step S1 is obtained is placed in the β CD-PAA aqueous solution that concentration is 8mg/mL, lucifuge, magnetic at room temperature
Power is stirred 4 hours, and microballoon is washed with deionized 3 times after centrifugation.
S3, the microballoon that step S2 is obtained is put in drying at room temperature 13h in vacuum drying chamber, dried microballoon is placed in purple
Exposure-processed 15min under outer light, obtains can be used as the improved silica microballoon β CD-PAA@SiO of HPLC chiral stationary phases2。
The diazo resin of the present embodiment is structure, wherein n=6 shown in formula (1).
Embodiment 4
The preparation method of the HPLC chiral stationary phases of the present embodiment, comprises the steps：
The activation process of S0, silicon dioxide microsphere：
The monodisperse silica microspheres that 2g average grain diameters are 1 μm are evenly dispersed in the salt that concentration is 0.12mol/L
30min is processed in aqueous acid, silicon dioxide microsphere is simultaneously washed to neutrality by centrifugation；
S1, the silicon dioxide microsphere after activation process is modified：
By the silicon dioxide microsphere after activation be placed in concentration be 12mg/mL the diazo resin aqueous solution in, at room temperature lucifuge,
Magnetic agitation 3 hours, microballoon is washed with deionized 3 times after centrifugation；
S2, the microballoon that step S1 is obtained is placed in the β CD-PAA aqueous solution that concentration is 12mg/mL, lucifuge, magnetic at room temperature
Power is stirred 3 hours, and microballoon is washed with deionized 3 times after centrifugation.
S3, the microballoon that step S2 is obtained is put in drying at room temperature 14h in vacuum drying chamber, dried microballoon is placed in purple
Exposure-processed 15min under outer light, obtains can be used as the improved silica microballoon β CD-PAA@SiO of HPLC chiral stationary phases2。
The diazo resin of the present embodiment is structure, wherein n=3 shown in formula (1).
Application examples 1
By the improved silica microballoon β CD-PAA@SiO that can be used as HPLC chiral stationary phases of the gained of example 12
It is 100mm to fill out length with packing column machine under 40Mpa pressure, in the stainless steel chromatographic column of a diameter of 4.6mm, using acetonitrile as flowing
Phase, under the flow velocity of 0.2mL/min, chiral drug metoprolol tartrate has carried out high performance liquid chromatography separation, and
Separate substance is detected under the conditions of the ultraviolet detection of 250nm wavelength, as shown in Figure 3, Fig. 3 shows with two testing result
Individual clearly crest, (the S)-enantiomer and (R)-enantiomer of this explanation metoprolol tartrate realize separation, therefore modification
Microballoon chiral stationary phase afterwards has good chiral separation effect as chromatographic column filler.If it should be noted that winestone
(the S)-enantiomer and (R)-enantiomer of sour metoprolol are not separated, then a crest will only occur in testing result.
Application examples 2
Example 1 can be used as the improved silica microballoon β CD-PAA@SiO of HPLC chiral stationary phases2In 40Mpa pressure
It is 100mm that lower packing column machine fills out length, in the stainless steel chromatographic column of a diameter of 4.6mm, with acetonitrile-TEAA (the two volume ratio
7:3) (TEAA is the mixed aqueous solution of acetic acid and triethylamine, and wherein concentration of the triethylamine in TEAA is the PH of 0.3%, TEAA
For 4) as mobile phase, under the flow velocity of 0.2mL/min, chiral medicine fenazil has carried out high performance liquid chromatography separation, and
Separate substance is detected under the conditions of the ultraviolet detection of 250nm wavelength, testing result is as shown in Figure 4.
Fig. 4 shows has two clearly crests, this (S)-enantiomer for illustrating chiral drug fenazil and (R)-mapping
The microballoon chiral stationary phase that body realizes after separation, therefore modification has good chiral separation effect as chromatographic column filler.
It should be noted that if (the S)-enantiomer of chiral drug fenazil and (R)-enantiomer are not separated, detection is tied
A crest will only occur in fruit.
Application examples 3
Example 1 can be used as the improved silica microballoon β CD-PAA@SiO of HPLC chiral stationary phases2In 40Mpa pressure
It is 100mm that lower packing column machine fills out length, in the stainless steel chromatographic column of a diameter of 4.6mm, with acetonitrile-TEAA (the two volume ratio
7:3) (TEAA is the mixed aqueous solution of acetic acid and triethylamine, and wherein concentration of the triethylamine in TEAA is the PH of 0.3%, TEAA
For 4) as mobile phase, under the flow velocity of 0.2mL/min, chiral medicine Dioxopromethazine has carried out high performance liquid chromatography separation,
And separate substance is detected under the conditions of the ultraviolet detection of 250nm wavelength, testing result is as shown in Figure 5.
Fig. 5 shown with two clearly crests, (the S)-enantiomer and (R) of this explanation chiral drug Dioxopromethazine-right
Reflect body and realize separation, therefore microballoon chiral stationary phase after modification as chromatographic column filler there is good chiral separation to imitate
Really.If it should be noted that (the S)-enantiomer of chiral drug Dioxopromethazine and (R)-enantiomer are not separated, examined
Surveying result a crest will only occurs.
Application examples 4
Example 1 can be used as the improved silica microballoon β CD-PAA@SiO of HPLC chiral stationary phases2In 40Mpa pressure
It is 100mm that lower packing column machine fills out length, in the stainless steel chromatographic column of a diameter of 4.6mm, with methyl alcohol-TEAA (7:3) (wherein
TEAA is 0.3%PH 4) used as mobile phase, under the flow velocity of 0.2mL/min, chiral medicine chlorpheniramine has carried out efficient liquid phase
Chromatographic isolation, and separate substance is detected under the conditions of the ultraviolet detection of 250nm wavelength, testing result is as shown in Figure 6.
Fig. 6 shows has two clearly crests, this (S)-enantiomer for illustrating chiral drug chlorpheniramine and (R)-mapping
The microballoon chiral stationary phase that body realizes after separation, therefore modification has good chiral separation effect as chromatographic column filler.
It should be noted that if (the S)-enantiomer of chiral drug chlorpheniramine and (R)-enantiomer are not separated, detection is tied
A crest will only occur in fruit.
Obviously, above-described embodiment is only intended to clearly illustrate example, and not to the restriction of implementation method.It is right
For those of ordinary skill in the art, can also make on the basis of the above description other multi-forms change or
Change.There is no need and unable to be exhaustive to all of implementation method.And the obvious change thus extended out or
Among changing still in the protection domain of the invention.The change or variation of form.There is no need and unable to all of reality
The mode of applying is exhaustive.And the obvious change thus extended out or change protection domain still in the invention
Among.
Claims (10)
1. a kind of preparation method of HPLC chiral stationary phases, it is characterised in that comprise the steps：
S1, silicon dioxide microsphere is dispersed in diazo resin solution, lucifuge is stirred 2-4 hours at room temperature, centrifugation
After wash microballoon with water；
S2, the product of step S1 is placed in beta-schardinger dextrin/polyacrylic acid composite solution, lucifuge is stirred 2-4 hours at room temperature,
Wash microballoon after centrifugation with water；
S3, will step S2 microballoon dry after be placed in exposed under UV light treatment 10-20min, obtain can be used as HPLC chiral solid
Determine the improved silica microballoon of phase.
2. the preparation method of HPLC chiral stationary phases according to claim 1, it is characterised in that the preparation method is also wrapped
Include：
S0, silicon dioxide microsphere is evenly dispersed in concentration be 0.08-0.12mol/L hydrochloric acid solution in carry out activation process
15-45min, washes microballoon to neutrality with water after centrifugation.
3. a kind of preparation method of HPLC chiral stationary phases according to claim 2, it is characterised in that the silica
Microballoon is the monodisperse particles of 1-3 μm of average grain diameter.
4. the preparation method of HPLC chiral stationary phases according to claim 3, it is characterised in that diazonium in the step S1
Resin solution is the diazo resin aqueous solution of 8-12mg/mL.
5. the preparation method of HPLC chiral stationary phases according to claim 4, it is characterised in that β-ring in the step S2
Dextrin/polyacrylic acid composite solution is the beta-schardinger dextrin/polyacrylic acid composite aqueous solution of 8-12mg/mL.
6. the preparation method of HPLC chiral stationary phases according to claim 5, it is characterised in that step S0, S1, S2
In to wash microballoon with water be microballoon to be washed with deionized 2-4 times.
7. the preparation method of HPLC chiral stationary phases according to claim 6, it is characterised in that microballoon in the step S3
Drying is that microballoon is put into vacuum tank to carry out drying at room temperature 12-14 hours.
8. the HPLC chiral stationary phases that a kind of claim 1-7 any one methods describeds are prepared.
9. application of the HPLC chiral stationary phases described in claim 8 in separating chiral material, it is characterised in that including following
Step：
During HPLC chiral stationary phase packing column machines are filled in into stainless steel chromatographic column, column temperature be 20~30 DEG C, flow velocity be
Under the conditions of 0.2mL/min, chiral material sample carries out high performance liquid chromatography separation.
10. application according to claim 9, it is characterised in that described chiral material sample be metoprolol tartrate,
Fenazil, Dioxopromethazine or chlorpheniramine.
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| CN112480028A (en) * | 2020-12-01 | 2021-03-12 | 中国人民解放军海军军医大学 | Method for chiral resolution of promethazine |
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