CN106732473A - A kind of method that rosin based high molecular chromatographic column for being esterified modification separates Gastrodin - Google Patents
A kind of method that rosin based high molecular chromatographic column for being esterified modification separates Gastrodin Download PDFInfo
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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
- B01D15/265—Adsorption chromatography
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- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
- B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
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- 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
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Abstract
The present invention discloses a kind of method that rosin based high molecular chromatographic column for being esterified modification separates Gastrodin, the method will be esterified the rosinyl polymer microsphere packing column machine wet method dress post preparative chromatography post of modification, HPLC separates Gastrodin and its methoxyphenyl β D glucopyranosides of derivative 4, Detection wavelength：220~280nm, 30 ± 10 DEG C of temperature, 0.3~1.0mL/min of flow velocity, you can realize the separation of Gastrodin and 4 methoxyphenyl β D glucopyranosides.The method separating degree is higher, and better than C18 posts, Environmental Safety, selectivity are high, with preferable application prospect.
Description
Technical field
The invention belongs to high performance liquid chromatography field, particularly a kind of rosin based high molecular chromatographic column for being esterified modification point
From the method for Gastrodin.
Background technology
The chemical structural formula of Gastrodin (Gastrodin)：
Gastrodin (Gastrodin) is the active ingredient of rare traditional Chinese medicine rhizoma Gastrodiae, with increase center and peripheral arterial blood
Pipe compliance, reduces peripheral vascular resistance, increases cardiovascular and cerebrovascular CBF, produces gentle antihypertensive effect, and to cardiac muscle cell, brain
Tissue has protective effect, while have calmness, hypnosis, analgesia, enhancing immune etc. and acting on, being clinically widely used in treatment head
The illnesss such as pain dizziness, extremity numbness, child convulsion, epilepsy, tic, lockjaw, it is evident in efficacy, and non-evident effect.Especially
In recent years, Gastrodin has in symptoms such as depression, Alzheimer disease, ascitic type liver cancer, osteoporosis, liver fibrosis, antimigraine
New progress, widens its field of medical applications significantly；Therefore the height that Gastrodin extracts yield is to weigh rhizoma Gastrodiae processing work
The mark of skill, researchs and develops significant to the filler of the selective separating effect of Gastrodin.
High performance liquid chromatography (High performance liquid chromatography, abbreviation HPLC) has following spy
Point：Speed is fast, and generally one sample of analysis can be completed in 15~30min；High resolution, can be to the material pair of otherness very little
As chiral material carries out separation analysis；Sensitivity is high, and UV-detector is up to 0.01ng, and fluorescence and electrochemical detector are reachable
0.1pg；Pillar Reusability, different compounds are can be separated with a root chromatogram column；Because it has used non-destructive detector,
After sample analysis, mobile phase can be removed, reclaim a small amount of preciousness medicine, be prepared by the purifying that also can be used for sample.
At present, on the report of Gastrodin high performance liquid chromatography separation method, we find the following document：
Huang jiao, Jiang Deng-jun.Determination of Gastrodin Contents in Wild
and Cultivated Gastrodia elata from Different Regions of Chongqing by RP-
HPLC.Medicinal Plant.2012,3(5)：42-44；Method is boiled using RP-HPLC color, C is used18Chromatographic column (150mm
× 4.6mm, 5 μm), with the phosphoric acid solution of acetonitrile -0.05% (3:97) it is mobile phase, flow velocity is 1mL/min, and column temperature is 30 DEG C, inspection
Survey wavelength 220nm.
Liu Yuhong, Yi Jinhai, RP-HPLC method determine free Gastrodin, the gloomy glycosides of Bali and total Gastrodin in rhizoma Gastrodiae simultaneously, in
Patent medicine, 2012,34 (1)：182-184；Method is boiled using RP-HPLC color, C is used18Chromatographic column (250mm × 4.6mm, 5 μm),
With the glacial acetic acid solution of methyl alcohol -1% as mobile phase, gradient elution, flow velocity is 0.8mL/min, and column temperature is 35 DEG C, Detection wavelength
270nm。
The method that rosin based high molecular chromatographic column on application esterification modification separates Gastrodin, does not up to the present appear in the newspapers
Road..
The content of the invention
It is an object of the invention to solve above-mentioned problem, there is provided a kind of rosin based high molecular chromatogram of esterification modification
The method of post separation Gastrodin, the method sensitivity is high, simple to operate, will not cause secondary dirt to medicine, health products, food
Dye.
To achieve the above object, the present invention provides following technical scheme：
A kind of method that rosin based high molecular chromatographic column for being esterified modification separates Gastrodin is high by the abietyl for being esterified modification
Molecule microballoon packing column machine wet method dress post preparative chromatography post, HPLC separates Gastrodin and its derivative 4- methoxyphenyl β-D- pyrroles
Glucopyranoside glycosides, Detection wavelength：220~280nm, 30 ± 10 DEG C of temperature, 0.3~1.0mL/min of flow velocity, you can realize Gastrodin
With the separation of 4- methoxyphenyl β-D- glucopyranosides.
Used as the further improvement of technical scheme, the rosinyl polymer microsphere of above-described esterification modification is spherical many
Porous materials, its particle diameter be 5~15 μm, 5~50mgKOH/g of acid number, average pore size be 10~15nm, specific surface area be 80~
150m2/g。
As the further improvement of technical scheme, the rosinyl polymer microsphere preparation method of the above esterification modification
For：It is that rosin based high molecular is micro- by maleic rosin acrylic acid glycol ester-(methyl) acryl acid-methyl methacrylate copolymer
Ball first carries out acylation reaction with acylating reagent, then carries out esterification with esterifying reagent, you can obtain described esterification modification
Rosinyl polymer microsphere.The system of maleic rosin acrylic acid glycol ester-(methyl) acryl acid-methyl methacrylate copolymer
Preparation Method, in patent " a kind of terpolymer of Abietyl-containing and preparation method thereof " (patent application of applicant's earlier application
Number：201010100733.4) it is open.
Used as the further improvement of technical scheme, above-described acylating reagent is oxalyl chloride or thionyl chloride.
As the further improvement of technical scheme, the one kind in above-described esterifying reagent methyl alcohol, ethylene glycol or glycerine.
Used as the further improvement of technical scheme, above-described acylation reaction is：By maleic rosin acrylic acid ethylene glycol
Ester-(methyl) acryl acid-methyl methacrylate copolymer is rosinyl polymer microsphere and solvent hexamethylene, n-hexane, heptan
One kind in alkane, acetone, octane, 1,4- dioxane is 1 in mass ratio：10~16 are placed in reaction vessel, in ice-water bath, stir
Under the conditions of mixing, to acylating reagent is added dropwise in reaction, rosin based high molecular is 1 with acylating reagent mass ratio：0.3~0.9, drip
Bi Hou, heating reflux reaction 4~8 hours under the conditions of 60~85 DEG C subtract the reaction system use that acylation reaction terminates to obtain
Pressure distilling apparatus, steams unnecessary solvent and acylating reagent, obtains acylated rosinyl polymer microsphere.
Used as the further improvement of technical scheme, above-described esterification is：Just it is acylated rosin based high molecular micro-
One kind in ball and solvent hexamethylene, n-hexane, heptane, acetone, octane, 1,4- dioxane is 1 according to mass ratio：7~13
Add in reactor, be subsequently adding esterifying reagent, the mass ratio for being acylated rosinyl polymer microsphere and esterifying reagent is 1：0.2~
3, reaction is stirred at reflux under the conditions of 60~80 DEG C 1~4 hour, warm water washing microballoon to PH=6~7, ethanolic extraction 10~
50h, adds the boiling of distilled water agitating heating to remove the small-molecule substances such as ethanol, you can obtain the abietyl of described esterification modification
Polymer microsphere.
Used as the further improvement of technical scheme, above-described wet method dress post is the rosin based high molecular that will be esterified modification
Microballoon is mixed in the one kind in methyl alcohol, acetonitrile or chloroform, and ultrasonic disperse is uniform, and equal slurries are filled in sky chromatographic column, with dress post
Machine constant pressure pump loads 120~200min under 3000~3500psi pressure, after post flattens weighing apparatus, by chromatographic column from packing column machine
Remove, load onto column cap, the rosin based high molecular chromatographic column of esterification modification is obtained.
Compared with prior art, beneficial effects of the present invention are：
1. the rosinyl polymer microsphere of the esterification modification that the present invention is prepared, dilation is small, specific surface area is big, can be used for
The active ingredient of separating natural product, and can using in organic solvent, will not because of expansion saboteur's microballoon netted knot
Structure, causes recognition capability to be lost.
2. the present invention is to be esterified the abietyl high score that the esterification that the rosinyl polymer microsphere of modification is prepared as filler is modified
Sub- chromatographic column, because rich in pore structure not of uniform size, having the advantages that permeability is good, back pressure is low, efficient and high flux, higher
Flow velocity and pressure under, there is not the phenomenon of be collapsed, can be used under high flow rate.
3. chromatographic column used in the present invention, has preferably to Gastrodin and 4- methoxyphenyl β-D- glucopyranosides
Separating effect；In C18Separating degree on post is 0.37, and the separating degree in patent chromatographic column of the present invention is more than 1.97.
4. the rosin based high molecular of esterification modification used in the present invention is obtained by Material synthesis of natural products, safe nothing
Poison, high mechanical strength.
5. chromatographic column used in the present invention, sensitivity is high, simple to operate.
Brief description of the drawings
Fig. 1 C18Post is to Gastrodin and the high performance liquid chromatography of 4- methoxyphenyl β-D- glucopyranoside mixed solutions
Spectrogram；
Fig. 2 is the embodiment of the present invention 4 to Gastrodin and the height of 4- methoxyphenyl β-D- glucopyranoside mixed solutions
Effect liquid phase chromatogram spectrogram；
Fig. 3 is the embodiment of the present invention 5 to Gastrodin and the height of 4- methoxyphenyl β-D- glucopyranoside mixed solutions
Effect liquid phase chromatogram spectrogram；
Fig. 4 is the embodiment of the present invention 6 to Gastrodin and the height of 4- methoxyphenyl β-D- glucopyranoside mixed solutions
Effect liquid phase chromatogram spectrogram
Specific embodiment
With reference to embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited to
The scope that embodiment is represented.
It is esterified the preparation of the rosinyl polymer microsphere of modification：
Referred to as " abietyl is high for maleic rosin acrylic acid glycol ester-(methyl) acryl acid-methyl methacrylate copolymer
Molecule microballoon "
Embodiment 1
Rosinyl polymer microsphere 5g will be weighed and 78g hexamethylenes will be measured and be placed in reaction vessel, the two mass ratio will be 1:
15.6, under ice-water bath, stirring condition, to acylating reagent 2.55g oxalyl chlorides are added dropwise in reaction, rosin based high molecular is tried with acylated
Agent mass ratio is 1:0.51, after completion of dropping, be heated to reflux 80 DEG C and react 4 hours, then by acylation reaction terminate to obtain it is anti-
Answer system to use vacuum distillation apparatus, steam unnecessary hexamethylene and acylating reagent, obtain acylated rosinyl polymer microsphere.
Obtain being acylated in rosinyl polymer microsphere and solvent hexamethylene 39g addition reactors by foregoing, the two mass ratio
It is 1:7.8, esterifying reagent 4.75g methyl alcohol is subsequently adding, the mass ratio for being acylated rosinyl polymer microsphere and esterifying reagent is 1:
0.95, it is stirred at reflux 60 DEG C and reacts 1 hour, warm water washs microballoon to PH=6.0, ethanolic extraction 10h, adds distilled water stirring to add
Heat boiling removes the small-molecule substances such as ethanol, you can obtain the rosinyl polymer microsphere of described esterification modification.
Analyze after testing, the rosinyl polymer microsphere of the esterification modification that the present embodiment is obtained, the macromolecule after modification is micro-
Ball is spherical porous material, and its particle diameter is 5~15 μm, and 5~50mgKOH/g of acid number, average pore size is 10~15nm, specific surface area
It is 80~150m2/g。
Embodiment 2：
Isosorbide-5-Nitrae-dioxane that rosinyl polymer microsphere 10g will be weighed and 104g is measured is placed in reaction vessel, the two
Mass ratio is 1:10.4, under ice-water bath, stirring condition, to dropwise addition acylating reagent 5.1g oxalyl chlorides, abietyl high score in reaction
Son is 1 with acylating reagent mass ratio:0.51, after completion of dropping, it is heated to 70 DEG C and reacts 6 hours, then acylation reaction is terminated
The reaction system for obtaining uses vacuum distillation apparatus, steams unnecessary Isosorbide-5-Nitrae-dioxane and acylating reagent, obtains the pine of acylation
Perfume base polymer microsphere.
Obtain being acylated in rosinyl polymer microsphere and solvent Isosorbide-5-Nitrae-dioxane 100g addition reactors by foregoing, the two
Mass ratio is 1:10, it is subsequently adding the quality of esterifying reagent 9.93g ethylene glycol, acylated rosinyl polymer microsphere and esterifying reagent
Than being 1:0.99, stir 70 DEG C and react 3 hours, warm water washs microballoon to PH=6.5, ethanolic extraction 24h, adds distilled water stirring
Ebuillition of heated removes the small-molecule substances such as ethanol, you can obtain the rosinyl polymer microsphere of described esterification modification.
Analyze after testing, the rosinyl polymer microsphere of the esterification modification that the present embodiment is obtained, the macromolecule after modification is micro-
Ball is spherical porous material, and its particle diameter is 5~15 μm, and 5~50mgKOH/g of acid number, average pore size is 10~15nm, specific surface area
It is 80~150m2/g。
Embodiment 3：
Rosinyl polymer microsphere 5g will be weighed and 78.8g acetone will be measured and be placed in reaction vessel, the two mass ratio will be 1:
15.76, under ice-water bath, stirring condition, to dropwise addition acylating reagent 4.01g thionyl chlorides, rosin based high molecular and acyl in reaction
It is 1 to change reagent quality ratio:0.8, after completion of dropping, it is heated to reflux 60 DEG C and reacts 8 hours, acylation reaction is then terminated what is obtained
Reaction system uses vacuum distillation apparatus, steams unnecessary acetone and acylating reagent, obtains acylated rosinyl polymer microsphere.
Obtain being acylated in rosinyl polymer microsphere and solvent acetone 59.1g addition reactors by foregoing, the two mass ratio
It is 1:11.82, esterifying reagent 4.85g glycerine is subsequently adding, the mass ratio for being acylated rosinyl polymer microsphere and esterifying reagent is
1:0.97, it is stirred at reflux 80 DEG C and reacts 4 hours, warm water washs microballoon to PH=7.0, ethanolic extraction 36h, adds distilled water stirring
Ebuillition of heated removes the small-molecule substances such as ethanol, you can obtain the rosinyl polymer microsphere of described esterification modification.
Analyze after testing, the rosinyl polymer microsphere of the esterification modification that the present embodiment is obtained, the macromolecule after modification is micro-
Ball is spherical porous material, and its particle diameter is 5~15 μm, and 5~50mgKOH/g of acid number, average pore size is 10~15nm, specific surface area
It is 80~150m2/g。
Embodiment 4
Rosinyl polymer microsphere 5g will be weighed and 60g n-hexanes will be measured and be placed in reaction vessel, the two mass ratio will be 1:
12, under ice-water bath, stirring condition, to dropwise addition acylating reagent 1.5g oxalyl chlorides, rosin based high molecular and acylating reagent in reaction
Mass ratio is 1:0.3, after completion of dropping, it is heated to reflux 85 DEG C and reacts 5 hours, then acylation reaction is terminated the reactant for obtaining
System uses vacuum distillation apparatus, steams unnecessary n-hexane and acylating reagent, obtains acylated rosinyl polymer microsphere.
Obtain being acylated in rosinyl polymer microsphere and solvent hexane 45g addition reactors by foregoing, the two mass ratio
It is 1:9, esterifying reagent 10g methyl alcohol is subsequently adding, the mass ratio for being acylated rosinyl polymer microsphere and esterifying reagent is 1:2, stir
Mix 65 DEG C of backflow to react 2 hours, warm water washs microballoon to PH=6.0, ethanolic extraction 50h, add the boiling of distilled water agitating heating
Remove the small-molecule substances such as ethanol, you can obtain the rosinyl polymer microsphere of described esterification modification.
Analyze after testing, the rosinyl polymer microsphere of the esterification modification that the present embodiment is obtained, the macromolecule after modification is micro-
Ball is spherical porous material, and its particle diameter is 5~15 μm, and 5~50mgKOH/g of acid number, average pore size is 10~15nm, specific surface area
It is 80~150m2/g。
Embodiment 5：
The heptane that rosinyl polymer microsphere 10g will be weighed and 140g is measured is placed in reaction vessel, and the two mass ratio is
1:14, under ice-water bath, stirring condition, to acylating reagent 7.0g oxalyl chlorides are added dropwise in reaction, rosin based high molecular is tried with acylated
Agent mass ratio is 1:0.7, after completion of dropping, it is heated to 65 DEG C and reacts 7 hours, then acylation reaction is terminated the reactant for obtaining
System uses vacuum distillation apparatus, steams unnecessary heptane and acylating reagent, obtains acylated rosinyl polymer microsphere.
Obtain being acylated in rosinyl polymer microsphere and solvent heptane 130g addition reactors by foregoing, the two mass ratio is
1:13, esterifying reagent 30g ethylene glycol is subsequently adding, the mass ratio for being acylated rosinyl polymer microsphere and esterifying reagent is 1:3, stir
Mix 75 DEG C to react 3 hours, warm water washs microballoon to PH=6.5, ethanolic extraction 30h, add the boiling of distilled water agitating heating to remove
The small-molecule substances such as ethanol, you can obtain the rosinyl polymer microsphere of described esterification modification.
Analyze after testing, the rosinyl polymer microsphere of the esterification modification that the present embodiment is obtained, the macromolecule after modification is micro-
Ball is spherical porous material, and its particle diameter is 5~15 μm, and 5~50mgKOH/g of acid number, average pore size is 10~15nm, specific surface area
It is 80~150m2/g。
Embodiment 6：
Rosinyl polymer microsphere 5g will be weighed and 55g octanes will be measured and be placed in reaction vessel, the two mass ratio will be 1:11,
Under ice-water bath, stirring condition, to dropwise addition acylating reagent 3.0g thionyl chlorides, rosin based high molecular and acylating reagent matter in reaction
Amount is than being 1:0.6, after completion of dropping, it is heated to reflux 75 DEG C and reacts 8 hours, then acylation reaction is terminated the reaction system for obtaining
Using vacuum distillation apparatus, unnecessary octane and acylating reagent is steamed, obtain acylated rosinyl polymer microsphere.
Obtain being acylated in rosinyl polymer microsphere and solvent octane 55.0g addition reactors by foregoing, the two mass ratio
It is 1:11, esterifying reagent 1g glycerine is subsequently adding, the mass ratio for being acylated rosinyl polymer microsphere and esterifying reagent is 1:0.2,
It is stirred at reflux 65 DEG C to react 4 hours, warm water washs microballoon to PH=7.0, ethanolic extraction 40h, adds the boiling of distilled water agitating heating
Rise the small-molecule substances such as removing ethanol, you can obtain the rosinyl polymer microsphere of described esterification modification.
Analyze after testing, the rosinyl polymer microsphere of the esterification modification that the present embodiment is obtained, the macromolecule after modification is micro-
Ball is spherical porous material, and its particle diameter is 5~15 μm, and 5~50mgKOH/g of acid number, average pore size is 10~15nm, specific surface area
It is 80~150m2/g。
Using the method that the rosin based high molecular chromatographic column of esterification modification separates Gastrodin
Embodiment 7：
A kind of method that rosin based high molecular chromatographic column for being esterified modification separates Gastrodin, is carried out in accordance with the following steps：
(1) prepared by the rosin based high molecular chromatographic column of esterification modification：The rosin based high molecular that just esterification of embodiment 1 is modified
Microballoon is mixed in methyl alcohol, and ultrasonic disperse is uniform, equal slurries is filled in sky chromatographic column, with packing column machine constant pressure pump in 3000psi
120min is loaded under pressure, after post flattens weighing apparatus, chromatographic column is removed from packing column machine, load onto column cap, esterification modification is obtained
Rosin based high molecular chromatographic column.The rosin based high molecular chromatographic column methyl alcohol of the esterification modification of gained is rinsed to baseline balance again
Can sample introduction.
(2) sample solution is prepared：Appropriate Gastrodin, 4- methoxyphenyl β-D- glucopyranosides are taken, are dissolved with methyl alcohol,
Every 1L is configured to containing Gastrodin 2.5 × 10-4The Gastrodin of mol and 4- methoxyphenyl β-D- glucopyranoside mixed solutions,
Sample introduction；
(3) setup parameter：The rosin based high molecular chromatographic column that modification will be esterified accesses liquid chromatograph, sets liquid chromatogram
The flow rate of mobile phase of instrument is 0.3mL/min, Detection wavelength 220nm, 25 DEG C of column oven；
(4) separate：Starting sampling valve makes methyl alcohol bring in the rosin based high molecular chromatographic column that esterification is modified sample into, realizes
The separation of Gastrodin, acquired results as shown in Fig. 2 there is Gastrodin peak in retention time 11.06min, in retention time
Occurs 4- methoxyphenyl β-D- glucopyranosides peak during 12.39min, separating degree is 2.31.
Embodiment 8：
A kind of method that rosin based high molecular chromatographic column for being esterified modification separates Gastrodin, is carried out in accordance with the following steps：
(1) prepared by the rosin based high molecular chromatographic column of esterification modification：The rosin based high molecular that just esterification of embodiment 2 is modified
Microballoon is mixed in acetonitrile, and ultrasonic disperse is uniform, equal slurries is filled in sky chromatographic column, with packing column machine constant pressure pump in 3300psi
160min is loaded under pressure, after post flattens weighing apparatus, chromatographic column is removed from packing column machine, load onto column cap, esterification modification is obtained
Rosin based high molecular chromatographic column.The rosin based high molecular chromatographic column methyl alcohol of the esterification modification of gained is rinsed to baseline balance again
Can sample introduction.
(2) sample solution is prepared：Appropriate Gastrodin, 4- methoxyphenyl β-D- glucopyranosides are taken, are dissolved with methyl alcohol,
Every 1L is configured to containing Gastrodin 2.5 × 10-4The Gastrodin of mol and 4- methoxyphenyl β-D- glucopyranoside mixed solutions,
Sample introduction；
(2) setup parameter：The rosin based high molecular chromatographic column that modification will be esterified accesses liquid chromatograph, sets liquid chromatogram
The flow rate of mobile phase of instrument is 0.6mL/min, Detection wavelength 250nm, 35 DEG C of column oven；
(3) separate：Starting sampling valve makes methyl alcohol bring in the rosin based high molecular chromatographic column that esterification is modified sample into, realizes
The separation of Gastrodin, acquired results as shown in figure 3, there is Gastrodin peak in retention time 11.22min, in retention time
Occurs 4- methoxyphenyl β-D- glucopyranosides peak during 12.57min, separating degree is 1.97.
Embodiment 9：
A kind of method that rosin based high molecular chromatographic column for being esterified modification separates Gastrodin, is carried out in accordance with the following steps：
(1) prepared by the rosin based high molecular chromatographic column of esterification modification：The rosin based high molecular that just esterification of embodiment 3 is modified
Microballoon is mixed in chloroform, and ultrasonic disperse is uniform, equal slurries is filled in sky chromatographic column, with packing column machine constant pressure pump in 3500psi
200min is loaded under pressure, after post flattens weighing apparatus, chromatographic column is removed from packing column machine, load onto column cap, esterification modification is obtained
Rosin based high molecular chromatographic column.The rosin based high molecular chromatographic column methyl alcohol of the esterification modification of gained is rinsed to baseline balance again
Can sample introduction.
(2) sample solution is prepared：Appropriate Gastrodin, 4- methoxyphenyl β-D- glucopyranosides are taken, are dissolved with methyl alcohol,
Every 1L is configured to containing Gastrodin 2.5 × 10-4The Gastrodin of mol and 4- methoxyphenyl β-D- glucopyranoside mixed solutions,
Sample introduction；
(3) setup parameter：The rosin based high molecular chromatographic column that modification will be esterified accesses liquid chromatograph, sets liquid chromatogram
The flow rate of mobile phase of instrument is 1.0mL/min, Detection wavelength 270nm, 30 DEG C of column oven；
(4) separate：Starting sampling valve makes methyl alcohol bring in the rosin based high molecular chromatographic column that esterification is modified sample into, realizes
The separation of Gastrodin, acquired results as shown in figure 4, there is Gastrodin peak in retention time 11.18min, in retention time
Occurs 4- methoxyphenyl β-D- glucopyranosides peak during 12.43min, separating degree is 2.17.
Likewise, applicant carries out rhizoma Gastrodiae to the rosinyl polymer microsphere for implementing the preparation-obtained esterification modifications of 4-6
The separation of element and 4- methoxyphenyl β-D- glucopyranosides, separating degree can reach more than 1.97.
Claims (8)
1. a kind of method that rosin based high molecular chromatographic column for being esterified modification separates Gastrodin, it is characterised in that:Esterification is modified
Rosinyl polymer microsphere packing column machine wet method dress post preparative chromatography post, HPLC separate Gastrodin and its derivative 4- methoxyl groups
Phenyl β-D- glucopyranosides, Detection wavelength：220~280nm, 30 ± 10 DEG C of temperature, 0.3~1.0mL/min of flow velocity, you can
Realize the separation of Gastrodin and 4- methoxyphenyl β-D- glucopyranosides.
2. the method that a kind of rosin based high molecular chromatographic column for being esterified modification according to claim 1 separates Gastrodin, its
It is characterised by：The rosinyl polymer microsphere of described esterification modification is spherical porous material, and its particle diameter is 5~15 μm, acid number 5
~50mgKOH/g, average pore size is 10~15nm, and specific surface area is 80~150m2/g。
3. the method that a kind of rosin based high molecular chromatographic column for being esterified modification according to claim 1 and 2 separates Gastrodin,
It is characterized in that：It is described esterification modification rosinyl polymer microsphere preparation method be：By maleic rosin acrylic acid glycol ester-
(methyl) acryl acid-methyl methacrylate copolymer is that rosinyl polymer microsphere first carries out acylation reaction with acylating reagent,
Esterification is carried out with esterifying reagent, you can obtain the rosinyl polymer microsphere of described esterification modification again.
4. the method that a kind of rosin based high molecular chromatographic column for being esterified modification according to claim 3 separates Gastrodin, its
It is characterised by：Described acylating reagent is oxalyl chloride or thionyl chloride.
5. the method that a kind of rosin based high molecular chromatographic column for being esterified modification according to claim 3 separates Gastrodin, its
It is characterised by：One kind in described esterifying reagent methyl alcohol, ethylene glycol or glycerine.
6. the method that a kind of rosin based high molecular chromatographic column of esterification modification according to claim 4 or 5 separates Gastrodin,
It is characterized in that：Described acylation reaction is：By maleic rosin acrylic acid glycol ester-(methyl) acrylic acid-methacrylic acid
During methyl terpolymer is rosinyl polymer microsphere and solvent hexamethylene, n-hexane, heptane, acetone, octane, 1,4- dioxane
It is a kind of be in mass ratio 1：10~16 are placed in reaction vessel, under ice-water bath, stirring condition, examination are acylated to being added dropwise in reaction
Agent, rosin based high molecular is 1 with acylating reagent mass ratio：0.3~0.9, after completion of dropping, heated back under the conditions of 60~85 DEG C
Stream reaction 4~8 hours, the reaction system that acylation reaction terminates to obtain is used into vacuum distillation apparatus, steam unnecessary solvent and
Acylating reagent, obtains acylated rosinyl polymer microsphere.
7. the method that a kind of rosin based high molecular chromatographic column for being esterified modification according to claim 6 separates Gastrodin, its
It is characterised by：Described esterification is：Will acylated rosinyl polymer microsphere and solvent hexamethylene, n-hexane, heptane, third
One kind in ketone, octane, 1,4- dioxane is 1 according to mass ratio：In 7~13 addition reactors, esterifying reagent is subsequently adding,
The mass ratio of acylated rosinyl polymer microsphere and esterifying reagent is 1：0.2~3, reaction is stirred at reflux under the conditions of 60~80 DEG C
1~4 hour, warm water washed microballoon to PH=6~7, and 10~50h of ethanolic extraction adds the boiling of distilled water agitating heating to remove second
The small-molecule substances such as alcohol, you can obtain the rosinyl polymer microsphere of described esterification modification.
8. the method that a kind of rosin based high molecular chromatographic column for being esterified modification according to claim 1 and 2 separates Gastrodin,
It is characterized in that：Described wet method dress post is that the rosinyl polymer microsphere for being esterified modification is mixed in methyl alcohol, acetonitrile or chloroform
One kind, ultrasonic disperse is uniform, and equal slurries are filled in sky chromatographic column, is pressed in 3000~3500psi with packing column machine constant pressure pump
120~200min is loaded under power, after post flattens weighing apparatus, chromatographic column is removed from packing column machine, load onto column cap, esterification modification is obtained
Rosin based high molecular chromatographic column.
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| CN109900846A (en) * | 2019-03-29 | 2019-06-18 | 湖北民族大学 | It is a kind of to apply SiO2The method that@rosin based high molecular chromatographic column separates Gastrodin and its derivative |
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