CN108484557A - A method of extracting isofraxidin from wilsonii - Google Patents
A method of extracting isofraxidin from wilsonii Download PDFInfo
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- CN108484557A CN108484557A CN201810620030.0A CN201810620030A CN108484557A CN 108484557 A CN108484557 A CN 108484557A CN 201810620030 A CN201810620030 A CN 201810620030A CN 108484557 A CN108484557 A CN 108484557A
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- isofraxidin
- exchange resin
- ion exchange
- type ion
- extraction
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/06—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2
- C07D311/08—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring
- C07D311/16—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring substituted in position 7
Abstract
The method that the present invention provides a kind of to extract isofraxidin from wilsonii.The present invention is completely converted into free state isofraxidin, to increase the content of isofraxidin in extract liquor by the extraction in combination hydrolysis in situ, the glycosides compound that isofraxidin can be made to be combined into glucose, i.e. reference state isofraxidin；Wherein, the H+Type ion exchange resin is the hydrolysis for promoting reference state isofraxidin as catalyst, and the chlorinated solvents are to extract free state isofraxidin as extractant.Therefore, during extraction in combination hydrolysis in situ, the hydrolysis of reference state isofraxidin and the extraction of free state isofraxidin are completed at the same time.Meanwhile extraction in combination hydrolysis in situ method provided by the invention efficiently avoids hydrolysis and extracts the lengthy and tedious of two steps operation again, saves the usage amount of operational sequence and solvent, production cost is low, and isofraxidin yield is high.
Description
Technical field
The present invention relates to compound extractive technique fields, and in particular to a kind of side extracting isofraxidin from wilsonii
Method.
Background technology
Wilsonii is distributed widely in the multiple provinces in China and area, is important one of the Chinese herbal medicine in China.Wilsonii has
The effect of replenishing qi to invigorate the spleen, tonifying the kidney to relieve mental strain can enhance body nonspecific defense ability, except with immunological regulation, antitumor, anti-ageing
Always, outside Antiradiation injury and fatigue-resisting function, the diseases such as angiocardiopathy, diabetes and neurasthenia can also be treated.Wilsonii
Root and stem portion contain various active chemical composition, wherein isofraxidin and phenol glycosides compound is especially prominent.Isofraxidin
For one of active constituent important in wilsonii, there are a variety of pharmacology such as calm, anti-inflammatory, antibacterial, antitumor and prevention angiocarpy
The height of activity, content is one of the index for evaluating wilsonii quality good or not.
Currently, the method for isofraxidin is typically directly to extract extracting solution after extracting wilsonii in extraction wilsonii
Concentration is taken, then is detached with macroporous absorbent resin or silica gel.Such methods have the disadvantage that：It is free except existing in wilsonii
Outside state isofraxidin, there is also reference state isofraxidins；If reference state isofraxidin is not extracted can cause it is about 30% different
The loss of piperazine skin pyridine, meanwhile, resin or the separatory organic solvent recycling load of silica gel are big, cannot reuse, cause cost
With the waste on energy.
Invention content
The method that the purpose of the present invention is to provide a kind of to extract isofraxidin from wilsonii, method provided by the invention
The yield of isofraxidin is high, and at low cost.
In order to achieve the above-mentioned object of the invention, the present invention provides following technical scheme：
The method that the present invention provides a kind of to extract isofraxidin from wilsonii, includes the following steps：
Wilsonii is extracted using the first ethanol water, obtains extracting solution；
By the extracting solution and H+Extraction in combination hydrolysis in situ is carried out after type ion exchange resin and chlorinated solvents mixing, is extracted
Take liquid；
The organic phase in the extract liquor is isolated, the organic phase is concentrated to dryness, using the second ethanol water pair
Resulting material is dissolved, and lysate is obtained；
The lysate is enriched with using macroporous absorbent resin, using third ethanol water to the macropore after enrichment
Absorption resin is eluted, and the solvent in removal gained eluent obtains isofraxidin.
Preferably, the volumn concentration of first ethanol water is 60~90%；Second ethanol water
Volumn concentration be 20~30%；The volumn concentration of the third ethanol water is 75~90%.
Preferably, in terms of the absolute dry mass of the wilsonii, the mass ratio of the wilsonii and the first ethanol water is
1：(7~15).
Preferably, the temperature of the extraction is 70~90 DEG C, and the time is 1~5h.
Preferably, the H+Type ion exchange resin is pre-processed before use, the pretreated method include with
Lower step：
(1) use ethyl alcohol to H+Type ion exchange resin carries out immersion treatment, by gained H+Type ion exchange resin carries out water
It washes；
(2) H for using base reagent to obtain the step (1)+Type ion exchange resin carries out immersion treatment, by gained H+
Type ion exchange resin is washed to neutrality；
(3) H for using acid reagent to obtain the step (2)+Type ion exchange resin carries out immersion treatment, by gained H+
Type ion exchange resin is washed to neutrality；
(4) H for obtaining the step (3)+Type ion exchange resin carries out the place in step (2) and step (3) successively
Reason, obtains pretreated H+Type ion exchange resin is spare.
Preferably, in terms of the absolute dry mass of the wilsonii, the H+Type ion exchange resin is wilsonii quality
0.25~5%.
Preferably, the chlorinated solvents include dichloromethane, chloroform, 1,1- dichloroethanes or 1,2- dichloroethanes.
Preferably, the chlorinated solvents and the volume ratio of extracting solution are (0.4~2.5)：1.
Preferably, the extraction in combination hydrolysis in situ carries out under sealing condition；The temperature of the extraction in combination hydrolysis in situ be 75~
95 DEG C, the time is 1~4h.
Preferably, it the enrichment and is eluted under sealing condition and carries out；In the enrichment process sample concentration be 0.2~
1.0mg/mL。
The method that the present invention provides a kind of to extract isofraxidin from wilsonii, using the first ethanol water to thorn five
Add and extract, obtains extracting solution；By the extracting solution and H+It is hydrolyzed after type ion exchange resin and chlorinated solvents mixing
Situ extracting obtains extract liquor；The organic phase in the extract liquor is isolated, the organic phase is concentrated to dryness, using second
Ethanol water dissolves resulting material, obtains lysate；The lysate is enriched with using macroporous absorbent resin,
The macroporous absorbent resin after enrichment is eluted using third ethanol water, the solvent in removal gained eluent obtains
Isofraxidin.The present invention is by the extraction in combination hydrolysis in situ, the glycoside chemical combination that isofraxidin can be made to be combined into glucose
Object, i.e. reference state isofraxidin, are completely converted into free state isofraxidin, to increase the content of isofraxidin in extract liquor；
Wherein, the H+Type ion exchange resin is the hydrolysis for promoting reference state isofraxidin as catalyst, and the chlorinated solvents are
Free state isofraxidin is extracted as extractant.Therefore, during extraction in combination hydrolysis in situ, the hydrolysis of reference state isofraxidin and
The extraction of free state isofraxidin is completed at the same time.Meanwhile extraction in combination hydrolysis in situ method provided by the invention is effectively kept away again
Exempt from hydrolysis and extracted the lengthy and tedious of two steps operation, saves the usage amount of operational sequence and solvent, production cost is low, and isofraxidin obtains
Rate is high.Embodiment the experimental results showed that, by the yield of isofraxidin in extracting solution be 100% in terms of, according to provided by the invention
The yield for the isofraxidin that method obtains is up to 142.74%, and purity is up to 34.2%.
Further, extraction in combination hydrolysis in situ, enrichment and elution process are carried out under sealing condition in the present invention, no dirt
Dye, solvent are recyclable；And the H+Type ion exchange resin can recycle after through regeneration treatment.This is into one
Step reduces production cost, is produced convenient for industrial scale.
Description of the drawings
Fig. 1 is the high-efficient liquid phase chromatogram of gained isofraxidin (a) and isofraxidin standard items (b) in embodiment 1.
Specific implementation mode
The method that the present invention provides a kind of to extract isofraxidin from wilsonii, includes the following steps：
Wilsonii is extracted using the first ethanol water, obtains extracting solution；
By the extracting solution and H+Extraction in combination hydrolysis in situ is carried out after type ion exchange resin and chlorinated solvents mixing, is extracted
Take liquid；
The organic phase in the extract liquor is isolated, the organic phase is concentrated to dryness, using the second ethanol water pair
Resulting material is dissolved, and lysate is obtained；
The lysate is enriched with using macroporous absorbent resin, using third ethanol water to the macropore after enrichment
Absorption resin is eluted, and the solvent in removal gained eluent obtains isofraxidin.
The present invention extracts wilsonii using the first ethanol water, obtains extracting solution.In the present invention, described
The volumn concentration of one ethanol water is preferably 60~90%, and more preferably 70~80%.In the present invention, with the thorn
The mass ratio of the absolute dry mass meter of slender acanthopanax, the wilsonii and the first ethanol water is preferably 1：(7~15), more preferably 1：
(9~12).
In the present invention, the wilsonii is preferably the Radix Acanthopanacis Senticosi root after natural air drying；In the present invention, the wilsonii
For rhizome after natural air drying, moisture content is 20% hereinafter, in not necessary any drying process processing before.In the present invention,
The wilsonii is preferably crushing processing using preceding, obtains the Manyprickle Acanthopanax Root that grain size is 60~100 mesh.
In the present invention, the temperature of the extraction is preferably 70~90 DEG C, more preferably 75~85 DEG C；In the reality of the present invention
It applies in example, the temperature of the extraction is specially the reflux temperature of first ethanol water.In the present invention, the extraction
Time is preferably 1~5h, more preferably 2~4h.In the present invention, in order to ensure fully to extract isofraxidin, the extraction
Number be preferably 2~4 times, more preferably 3 times.
After completing the extraction, gained system is preferably separated by solid-liquid separation by the present invention, using obtained liquid material as
Extracting solution.The present invention does not have the mode of the separation of solid and liquid special restriction, using side well known to those skilled in the art
Formula is specific as filtered.
The present invention extracts wilsonii using the first ethanol water, and raw material is not required to additionally dry directly use, energy
Consumption is few；And isofraxidin yield is high, fire safety evaluating hidden danger is low；Meanwhile first ethanol water can remove the thorn five
The larger ingredient of polarity in adding, such as polysaccharide and protein.
After obtaining extracting solution, the present invention is by the extracting solution and H+It is carried out after type ion exchange resin and chlorinated solvents mixing
Extraction in combination hydrolysis in situ obtains extract liquor.In the present invention, the chlorinated solvents and the volume ratio of extracting solution be preferably (0.4~
2.5)：1, more preferably (1~2)：1.In the present invention, the chlorinated solvents preferably include dichloromethane, chloroform, 1,1-
Dichloroethanes or 1,2- dichloroethanes.
In the present invention, in terms of the absolute dry mass of the wilsonii, the H+Type ion exchange resin is preferably wilsonii
The 0.25~5% of quality, more preferably 0.5~3%, most preferably 1~2%.The present invention is for the H+Type amberlite
The type of fat and source do not have special restriction, using the H of type known to those skilled in the art+The city of type ion exchange resin
Sell commodity.In the present invention, the H+The model of type ion exchange resin preferably include SK1B, 001*7, D061, D072,
110, D113, D151, D152 or WK11.
In the present invention, the H+The model and property of type ion exchange resin are as shown in the table：
H+The model and property of type ion exchange resin
In the present invention, the H+Type ion exchange resin is preferably pre-processed using preceding, the pretreated side
Method preferably includes following steps：
(1) use ethyl alcohol to H+Type ion exchange resin carries out immersion treatment, by gained H+Type ion exchange resin carries out water
It washes；
(2) H for using base reagent to obtain the step (1)+Type ion exchange resin carries out immersion treatment, by gained H+
Type ion exchange resin is washed to neutrality；
(3) H for using acid reagent to obtain the step (2)+Type ion exchange resin carries out immersion treatment, by gained H+
Type ion exchange resin is washed to neutrality；
(4) H for obtaining the step (3)+Type ion exchange resin carries out the place in step (2) and step (3) successively
Reason, obtains pretreated H+Type ion exchange resin is spare.
Present invention preferably employs ethyl alcohol to H+Type ion exchange resin carries out immersion treatment, by gained H+Type amberlite
Fat is washed.In the present invention, the time of the immersion treatment is preferably 10~14h, more preferably 12h.In the present invention,
Water used by the washing is preferably distilled water；The present invention, can will be at immersion for the not special restriction of the washing
Gained H after reason+Ethyl alcohol in type ion exchange resin is cleaned.
After the completion of carrying out immersion treatment and washing using ethyl alcohol, present invention preferably employs base reagents to gained H+Type ion is handed over
It changes resin and carries out immersion treatment, by gained H+Type ion exchange resin is washed to neutrality.In the present invention, the base reagent is preferred
For NaOH solution.The present invention does not have the concentration of the NaOH solution special restriction, using known to those skilled in the art
Concentration；In an embodiment of the present invention, the concentration of the NaOH solution is specially 0.1mol/L.In the present invention, institute
The time for stating immersion treatment is preferably 10~14h, more preferably 12h.In the present invention, water used by the washing is preferably
Distilled water；The present invention, can be by gained H after immersion treatment for the not special restriction of the washing+Type ion exchange resin
It is washed till neutrality.
After the completion of carrying out immersion treatment and washing using base reagent, present invention preferably employs acid reagents to gained H+Type ion
Exchanger resin carries out immersion treatment, by gained H+Type ion exchange resin is washed to neutrality.In the present invention, the acid reagent is excellent
It is selected as HCl solution.The present invention does not have the concentration of the HCl solution special restriction, using known to those skilled in the art
Concentration；In an embodiment of the present invention, the concentration of the HCl solution is specially 0.1mol/L.In the present invention, described
The time of immersion treatment is preferably 10~14h, more preferably 12h.In the present invention, water used by the washing preferably steams
Distilled water；The present invention, can be by gained H after immersion treatment for the not special restriction of the washing+Type ion exchange resin is washed
To neutrality.
After the completion of carrying out immersion treatment and washing using acid reagent, the present invention is preferably tried according to above-mentioned technical proposal using alkali
Agent is to gained H+Type ion exchange resin carries out immersion treatment, by gained H+Type ion exchange resin is washed to neutrality；Then according to
Above-mentioned technical proposal is using acid reagent to gained H+Type ion exchange resin carries out immersion treatment, by gained H+Type amberlite
Fat is washed to neutrality, obtains pretreated H+Type ion exchange resin is spare.
In the present invention, the H+Often contain a small amount of oligomer in type ion exchange resin commercial goods and does not participate in reaction
Monomer, also inorganic impurities such as iron, lead, copper, when it is contacted with other solution, above-mentioned substance will be transferred in solution,
Influence the quality of final product.Therefore, to the H+Type ion exchange resin, which carries out pretreatment, can prevent solubility therein miscellaneous
Matter is transferred in solution, using initial stage, pollutes the water quality of water outlet.
In the present invention, the extraction in combination hydrolysis in situ preferably carries out under sealing condition；The temperature of the extraction in combination hydrolysis in situ
Preferably 75~95 DEG C of degree, more preferably 80~90 DEG C, most preferably 85 DEG C；Time is preferably 1~4h, more preferably 2~3h.
By the extraction in combination hydrolysis in situ, the glycosides compound that isofraxidin can be made to be combined into glucose combines the present invention
State isofraxidin is completely converted into free state isofraxidin, to increase the content of isofraxidin in extract liquor；Wherein, the H+Type ion exchange resin is the hydrolysis for promoting reference state isofraxidin as catalyst, and the chlorinated solvents are as extractant
Extract free state isofraxidin.Therefore, during extraction in combination hydrolysis in situ, the hydrolysis of reference state isofraxidin and the different piperazine of free state
The extraction of skin pyridine is completed at the same time.Meanwhile extraction in combination hydrolysis in situ method provided by the invention efficiently avoid again hydrolysis with
The lengthy and tedious of two steps operation is extracted, the usage amount of operational sequence and solvent is saved, production cost is low, and isofraxidin yield is high.In addition,
The extraction in combination hydrolysis in situ carries out under sealing condition, pollution-free, and solvent is recyclable, the H+Type ion exchange resin
Through regeneration treatment (with the H after use+Type ion exchange resin is consistent using the pretreatment of preceding progress, no longer goes to live in the household of one's in-laws on getting married herein
State) it can recycle, it is produced convenient for industrial scale.
After obtaining extract liquor, the present invention isolates the organic phase in the extract liquor, and the organic phase is concentrated to dryness, is adopted
Resulting material is dissolved with the second ethanol water, obtains lysate.The present invention preferably by the extract liquor stratification,
To isolate organic phase.The present invention is for the not special restriction of the concentration, using well known to those skilled in the art dense
The technical solution of contracting.In the present invention, the volumn concentration of second ethanol water is preferably 20~30%, more
Preferably 23~27%.In the present invention, in terms of absolute dry mass, the organic phase is concentrated to dryness rear resulting material and the second ethyl alcohol
The mass ratio of aqueous solution is preferably 1：(25~35), more preferably 1：30.
After obtaining lysate, the present invention is enriched with the lysate using macroporous absorbent resin, using third ethyl alcohol
Aqueous solution elutes the macroporous absorbent resin after enrichment, and the solvent in removal gained eluent obtains isofraxidin.This hair
It is bright there is no special restriction for the type of the macroporous absorbent resin and source, using type known to those skilled in the art
The commercial goods of macroporous absorbent resin.In the present invention, the model of the macroporous absorbent resin preferably include HPD80,
HPD100, HPD300, HPD5000, AB-8, D101, HPD-D, DM-130, ADS-17, HPD826, ADS-7 or DA-201.
In the present invention, the model and property of the macroporous absorbent resin are as shown in the table：
The model and property of macroporous absorbent resin
In the present invention, the macroporous absorbent resin is pre-processed before use, and the pretreated method is preferably wrapped
Include following steps：
Macroporous absorbent resin is impregnated into 8~12h in the ethyl alcohol that volumn concentration is 90~95%, using distilled water pair
Macroporous absorbent resin after immersion is washed to remove remaining ethyl alcohol, the filtering of family name's funnel, and port grinding bottle seals 22~26h balances up for safekeeping
It is for use to obtain pretreated macroporous absorbent resin for moisture.
In the present invention, sample concentration is preferably 0.2~1.0mg/mL in the enrichment process, more preferably 0.3~
0.8mg/mL.In the present invention, the volumn concentration of the third ethanol water is preferably 75~90%, and more preferably 80
~85%.
In the present invention, the enrichment and elution preferably carry out under sealing condition, and pollution-free, solvent is recyclable to be made
With convenient for industrial scale production.The present invention does not have special restriction for the concrete operation method of the enrichment and elution, adopts
With the technical solution of enrichment and elution well known to those skilled in the art.Macroporous absorbent resin is preferably filled column by the present invention,
Under sealing condition, the lysate is enriched with using gained closing column, then uses the third ethanol water pair
Closing column after enrichment is eluted.In the present invention, the blade diameter length ratio of the closing column is preferably 1：(18~22), more preferably
1:20；The volume of the lysate is preferably close column volume 3~4 times, more preferably 3.5 times；The third ethyl alcohol is water-soluble
The volume of liquid is preferably close column volume 15~25 times, more preferably 20 times.
The method of solvent does not have special restriction in gained eluent after the present invention elutes removal, using this field skill
The technical solution of removal solvent known to art personnel is specific as being evaporated under reduced pressure.In the present invention, molten in gained eluent
Agent preferably carries out recycling, to reduce production cost.
After solvent in removal gained eluent, preferably resulting material is dried by the present invention, obtains isofraxidin.
In the present invention, the drying is preferably dried in vacuo；In the present invention, the vacuum drying temperature is preferably 35~45 DEG C,
More preferably 40 DEG C；The time is preferably 5~7h, more preferably 6h.
Below in conjunction with the embodiment in the present invention, the technical solution in the present invention is clearly and completely described.It is aobvious
So, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based on the reality in the present invention
Example is applied, every other embodiment obtained by those of ordinary skill in the art without making creative efforts all belongs to
In the scope of protection of the invention.
Embodiment 1
Radix Acanthopanacis Senticosi root after natural air drying is crushed into processing, the Manyprickle Acanthopanax Root that grain size is 70 mesh is obtained, weighs
10.0g Manyprickle Acanthopanax Roots are put into (in terms of absolute dry mass) in round-bottomed flask, then press the matter of Manyprickle Acanthopanax Root and ethanol water
Amount is than being 1：12 meters, are added the ethanol water that volumn concentration is 80%, and the refluxing extraction 2h in 85 DEG C of water-baths continues weight
It extracts again 2 times, merges resulting material after extraction, filtering, using obtained liquid material as extracting solution；
To H+Type ion exchange resin (model 001*7) is pre-processed, and specifically uses ethyl alcohol to H+Type ion exchange
Resin carries out immersion treatment 12h, by gained H+Type ion exchange resin is washed using distilled water；Using 0.1mol/L's
NaOH solution is to gained H+Type ion exchange resin carries out immersion treatment 12h, by gained H+Type ion exchange resin uses distilled water
It is washed till neutrality；Using the HCl solution of 0.1mol/L to gained H+Type ion exchange resin carries out immersion treatment 12h, by gained H+Type
Ion exchange resin is washed to neutrality using distillation；It repeats to carry out immersion treatment, distilled water using the NaOH solution of 0.1mol/L
It is washed till neutral and HCl solution using 0.1mol/L to carry out immersion treatment, distill the step for being washed to neutrality, after obtaining pretreatment
H+Type ion exchange resin；
By the extracting solution and pretreated H+Type ion exchange resin 1.0g and 30mL1, the mixing of 2- dichloroethanes,
Under sealing condition, reflux progress extraction in combination hydrolysis in situ 2h, obtains extract liquor in 90 DEG C of water-baths；
By the extract liquor stratification, organic phase is isolated, the organic phase is concentrated to dryness, then presses resulting material
The mass ratio of (in terms of absolute dry mass) and ethanol water is 1：It is water-soluble that the ethyl alcohol that volumn concentration is 25% is added in 30 meters
Liquid makes the organic phase be concentrated to dryness rear resulting material dissolving, obtains lysate；
Macroporous absorbent resin (model AB-8) is impregnated into 10h in the ethyl alcohol that volumn concentration is 95%, using steaming
Distilled water washs the macroporous absorbent resin after immersion to remove remaining ethyl alcohol, and the filtering of family name's funnel, port grinding bottle is sealed up for safekeeping puts down for 24 hours
The pretreated macroporous absorbent resin of gained is filled column by Hengshui point, and under sealing condition, column (blade diameter length ratio 1 is closed using gained：
20, volume 11.8mL) lysate is enriched with, sample concentration is 1.0mg/mL in the enrichment process；Then it adopts
The ethanol water for being 80% with volumn concentration (volume is close column volume 20 times) carries out the closing column after enrichment
Elution, vacuum distillation remove the solvent in gained eluent, obtain isofraxidin.
The content of isofraxidin in extracting solution is measured, yield is calculated；Meanwhile it calculating according to different obtained by method in embodiment 1
The yield of piperazine skin pyridine.The result shows that being to be obtained according to method in embodiment 1 in terms of 100% by the yield of isofraxidin in extracting solution
Isofraxidin yield be 128.68%.This explanation during extraction in combination hydrolysis in situ, isofraxidin and glucose in conjunction with and
At glycosides compound, i.e. reference state isofraxidin is converted into free state isofraxidin, to improve isofraxidin
Rate.To carrying out purity analysis according to isofraxidin obtained by method in embodiment 1, the results showed that, the purity of gained isofraxidin reaches
33.5%.Wherein, the content and purity of the isofraxidin by high performance liquid chromatography detection and are calculated, specifically
As shown in Figure 1.
According to H+Type ion exchange resin carries out pretreated method, and the H after situ extracting will be hydrolyzed+Type ion
Exchanger resin is regenerated.The result shows that by the H after regeneration+Type ion exchange resin is reused 4 times, and hydrolysis efficiency is first
The 86.52% of secondary hydrolysis efficiency.
Embodiment 2
Radix Acanthopanacis Senticosi root after natural air drying is crushed into processing, the Manyprickle Acanthopanax Root that grain size is 60 mesh is obtained, weighs
100.0g Manyprickle Acanthopanax Roots are put into (in terms of absolute dry mass) in round-bottomed flask, then press the matter of Manyprickle Acanthopanax Root and ethanol water
Amount is than being 1：15 meters, are added the ethanol water that volumn concentration is 85%, and the refluxing extraction 2.5h in 75 DEG C of water-baths continues
Repeat extraction 2 times, merges resulting material after extraction, filtering, using obtained liquid material as extracting solution；
To H+Type ion exchange resin (model 001*7) is pre-processed, and specifically uses ethyl alcohol to H+Type ion exchange
Resin carries out immersion treatment 12h, by gained H+Type ion exchange resin is washed using distilled water；Using 0.1mol/L's
NaOH solution is to gained H+Type ion exchange resin carries out immersion treatment 12h, by gained H+Type ion exchange resin uses distilled water
It is washed till neutrality；Using the HCl solution of 0.1mol/L to gained H+Type ion exchange resin carries out immersion treatment 12h, by gained H+Type
Ion exchange resin is washed to neutrality using distillation；It repeats to carry out immersion treatment, distilled water using the NaOH solution of 0.1mol/L
It is washed till neutral and HCl solution using 0.1mol/L to carry out immersion treatment, distill the step for being washed to neutrality, after obtaining pretreatment
H+Type ion exchange resin；
By the extracting solution and pretreated H+Type ion exchange resin 1.0g and 30mL1, the mixing of 2- dichloroethanes,
Under sealing condition, reflux progress extraction in combination hydrolysis in situ 3.5h, obtains extract liquor in 90 DEG C of water-baths；
By the extract liquor stratification, organic phase is isolated, the organic phase is concentrated to dryness, then presses resulting material
The mass ratio of (in terms of absolute dry mass) and ethanol water is 1：It is water-soluble that the ethyl alcohol that volumn concentration is 30% is added in 25 meters
Liquid makes the organic phase be concentrated to dryness rear resulting material dissolving, obtains lysate；
Macroporous absorbent resin (model AB-8) is impregnated into 10h in the ethyl alcohol that volumn concentration is 95%, using steaming
Distilled water washs the macroporous absorbent resin after immersion to remove remaining ethyl alcohol, and the filtering of family name's funnel, port grinding bottle is sealed up for safekeeping puts down for 24 hours
The pretreated macroporous absorbent resin of gained is filled column by Hengshui point, and under sealing condition, column (blade diameter length ratio 1 is closed using gained：
20, volume 11.8mL) lysate is enriched with, sample concentration is 1.0mg/mL in the enrichment process；Then it adopts
The ethanol water for being 90% with volumn concentration (volume is close column volume 20 times) carries out the closing column after enrichment
Elution, vacuum distillation remove the solvent in gained eluent, obtain isofraxidin.
The content of isofraxidin in extracting solution is measured, yield is calculated；Meanwhile it calculating according to different obtained by method in embodiment 2
The yield of piperazine skin pyridine.The result shows that being in terms of 100%, obtained by method in embodiment 2 by the yield of isofraxidin in extracting solution
The yield of isofraxidin is 142.74%.During extraction in combination hydrolysis in situ, isofraxidin is combined into this explanation with glucose
Glycosides compound, i.e. reference state isofraxidin is converted into free state isofraxidin, to improve the yield of isofraxidin.
To carrying out purity analysis according to isofraxidin obtained by method in embodiment 2, the results showed that, gained isofraxidin it is pure
It spends up to 34.2%.
According to H+Type ion exchange resin carries out pretreated method, and the H after situ extracting will be hydrolyzed+Type ion
Exchanger resin is regenerated.The result shows that by the H after regeneration+Type ion exchange resin is reused 4 times, and hydrolysis efficiency is first
The 88.22% of secondary hydrolysis efficiency.
Embodiment 3
Radix Acanthopanacis Senticosi root after natural air drying is crushed into processing, the Manyprickle Acanthopanax Root that grain size is 70 mesh is obtained, weighs
1000.0g Manyprickle Acanthopanax Roots are put into (in terms of absolute dry mass) in pilot scale round-bottomed flask, then press Manyprickle Acanthopanax Root and ethyl alcohol is water-soluble
The mass ratio of liquid is 1：The ethanol water that volumn concentration is 85%, the refluxing extraction in 90 DEG C of water-baths is added in 10 meters
2.5h continues to repeat extraction 2 times, merges resulting material after extraction, filtering, using obtained liquid material as extracting solution；
To H+Type ion exchange resin (model 001*7) is pre-processed, and specifically uses ethyl alcohol to H+Type ion exchange
Resin carries out immersion treatment 12h, by gained H+Type ion exchange resin is washed using distilled water；Using 0.1mol/L's
NaOH solution is to gained H+Type ion exchange resin carries out immersion treatment 12h, by gained H+Type ion exchange resin uses distilled water
It is washed till neutrality；Using the HCl solution of 0.1mol/L to gained H+Type ion exchange resin carries out immersion treatment 12h, by gained H+Type
Ion exchange resin is washed to neutrality using distillation；It repeats to carry out immersion treatment, distilled water using the NaOH solution of 0.1mol/L
It is washed till neutral and HCl solution using 0.1mol/L to carry out immersion treatment, distill the step for being washed to neutrality, after obtaining pretreatment
H+Type ion exchange resin；
By the extracting solution and pretreated H+Type ion exchange resin 1.0g and 30mL1, the mixing of 2- dichloroethanes,
Under sealing condition, reflux progress extraction in combination hydrolysis in situ 2h, obtains extract liquor in 80 DEG C of water-baths；
By the extract liquor stratification, organic phase is isolated, the organic phase is concentrated to dryness, then presses resulting material
The mass ratio of (in terms of absolute dry mass) and ethanol water is 1：It is water-soluble that the ethyl alcohol that volumn concentration is 20% is added in 30 meters
Liquid makes the organic phase be concentrated to dryness rear resulting material dissolving, obtains lysate；
Macroporous absorbent resin (model AB-8) is impregnated into 10h in the ethyl alcohol that volumn concentration is 95%, using steaming
Distilled water washs the macroporous absorbent resin after immersion to remove remaining ethyl alcohol, and the filtering of family name's funnel, port grinding bottle is sealed up for safekeeping puts down for 24 hours
The pretreated macroporous absorbent resin of gained is filled column by Hengshui point, and under sealing condition, column (blade diameter length ratio 1 is closed using gained：
20, volume 11.8mL) lysate is enriched with, sample concentration is 1.0mg/mL in the enrichment process；Then it adopts
The ethanol water for being 80% with volumn concentration (volume is close column volume 20 times) carries out the closing column after enrichment
Elution, vacuum distillation remove the solvent in gained eluent, obtain isofraxidin.
The content of isofraxidin in extracting solution is measured, yield is calculated；Meanwhile it calculating according to different obtained by method in embodiment 3
The yield of piperazine skin pyridine.The result shows that being in terms of 100%, obtained by method in embodiment 3 by the yield of isofraxidin in extracting solution
The yield of isofraxidin is 134.56%.During extraction in combination hydrolysis in situ, isofraxidin is combined into this explanation with glucose
Glycosides compound, i.e. reference state isofraxidin is converted into free state isofraxidin, to improve the yield of isofraxidin.
To carrying out purity analysis according to isofraxidin obtained by method in embodiment 3, the results showed that, gained isofraxidin it is pure
It spends up to 32.6%.
According to H+Type ion exchange resin carries out pretreated method, and the H after situ extracting will be hydrolyzed+Type ion
Exchanger resin is regenerated.The result shows that by the H after regeneration+Type ion exchange resin is reused 4 times, and hydrolysis efficiency is first
The 85.44% of secondary hydrolysis efficiency.
As seen from the above embodiment, the present invention can make isofraxidin and glucose knot by the extraction in combination hydrolysis in situ
Glycosides compound made of conjunction, i.e. reference state isofraxidin, are completely converted into free state isofraxidin, to increase in extract liquor
The content of isofraxidin；Wherein, the H+Type ion exchange resin is the hydrolysis for promoting reference state isofraxidin as catalyst,
The chlorinated solvents are to extract free state isofraxidin as extractant.Therefore, during extraction in combination hydrolysis in situ, reference state is different
The hydrolysis of piperazine skin pyridine and the extraction of free state isofraxidin are completed at the same time.Meanwhile extraction in combination hydrolysis in situ provided by the invention
Method efficiently avoids hydrolysis and extracts the lengthy and tedious of two steps operation again, saves the usage amount of operational sequence and solvent, is produced into
This is low, and isofraxidin yield is high.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of method for extracting isofraxidin from wilsonii, includes the following steps：
Wilsonii is extracted using the first ethanol water, obtains extracting solution；
By the extracting solution and H+Extraction in combination hydrolysis in situ is carried out after type ion exchange resin and chlorinated solvents mixing, obtains extract liquor；
The organic phase in the extract liquor is isolated, the organic phase is concentrated to dryness, using the second ethanol water to gained
Material is dissolved, and lysate is obtained；
The lysate is enriched with using macroporous absorbent resin, using third ethanol water to the macroporous absorption after enrichment
Resin is eluted, and the solvent in removal gained eluent obtains isofraxidin.
2. according to the method described in claim 1, it is characterized in that, the volumn concentration of first ethanol water is 60
~90%；The volumn concentration of second ethanol water is 20~30%；The volume hundred of the third ethanol water
It is 75~90% to divide content.
3. method according to claim 1 or 2, which is characterized in that in terms of the absolute dry mass of the wilsonii, the thorn five
It is 1 to add with the mass ratio of the first ethanol water：(7~15).
4. according to the method described in claim 3, it is characterized in that, the temperature of the extraction be 70~90 DEG C, the time be 1~
5h。
5. according to the method described in claim 1, it is characterized in that, the H+Type ion exchange resin is located in advance using preceding
Reason, the pretreated method include the following steps：
(1) use ethyl alcohol to H+Type ion exchange resin carries out immersion treatment, by gained H+Type ion exchange resin is washed；
(2) H for using base reagent to obtain the step (1)+Type ion exchange resin carries out immersion treatment, by gained H+Type from
Sub-exchange resin is washed to neutrality；
(3) H for using acid reagent to obtain the step (2)+Type ion exchange resin carries out immersion treatment, by gained H+Type from
Sub-exchange resin is washed to neutrality；
(4) H for obtaining the step (3)+Type ion exchange resin carries out the processing in step (2) and step (3) successively, obtains
To pretreated H+Type ion exchange resin is spare.
6. method according to claim 1 or 5, which is characterized in that in terms of the absolute dry mass of the wilsonii, the H+Type
Ion exchange resin is the 0.25~5% of wilsonii quality.
7. according to the method described in claim 1, it is characterized in that, the chlorinated solvents include dichloromethane, chloroform, 1,
1- dichloroethanes or 1,2- dichloroethanes.
8. method according to claim 1 or claim 7, which is characterized in that the chlorinated solvents and the volume ratio of extracting solution are
(0.4~2.5)：1.
9. according to the method described in claim 1, it is characterized in that, the extraction in combination hydrolysis in situ carries out under sealing condition；Institute
The temperature for stating extraction in combination hydrolysis in situ is 75~95 DEG C, and the time is 1~4h.
10. according to the method described in claim 1, it is characterized in that, the enrichment and being eluted under sealing condition and carrying out；It is described
Sample concentration is 0.2~1.0mg/mL in enrichment process.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113521803A (en) * | 2021-08-04 | 2021-10-22 | 佳木斯大学 | Eutectic solvent, preparation method thereof and application of eutectic solvent in extraction of isofraxidin in acanthopanax senticosus |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1450061A (en) * | 2002-04-10 | 2003-10-22 | 东北林业大学 | Process for extracting Isofraxidin |
| CN101851219A (en) * | 2010-05-27 | 2010-10-06 | 东北林业大学 | Method for preparing isofraxidin from acanthopanax senticosus rhizomes |
| CN101967504A (en) * | 2010-06-28 | 2011-02-09 | 南京泽朗农业发展有限公司 | Method for extracting isofraxidin |
-
2018
- 2018-06-15 CN CN201810620030.0A patent/CN108484557B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1450061A (en) * | 2002-04-10 | 2003-10-22 | 东北林业大学 | Process for extracting Isofraxidin |
| CN101851219A (en) * | 2010-05-27 | 2010-10-06 | 东北林业大学 | Method for preparing isofraxidin from acanthopanax senticosus rhizomes |
| CN101967504A (en) * | 2010-06-28 | 2011-02-09 | 南京泽朗农业发展有限公司 | Method for extracting isofraxidin |
Non-Patent Citations (3)
| Title |
|---|
| 杨磊，郝婧玮，刘洋，刘婷婷，祖元刚: "水解原位萃取分离刺五加异秦皮啶", 《中国现代应用药学》 * |
| 樊如强，金学英，胡荣，陈鹏，王建斌，傅宏征: "大孔树脂分离纯化刺五加中紫丁香苷、刺五加苷E和异嗪皮啶的工艺研究", 《中国现代应用药学》 * |
| 袁昕蓉，毕开顺，李强: "正交设计优化刺五加中游离异嗪皮啶", 《中药材》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113521803A (en) * | 2021-08-04 | 2021-10-22 | 佳木斯大学 | Eutectic solvent, preparation method thereof and application of eutectic solvent in extraction of isofraxidin in acanthopanax senticosus |
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