CN104844497B - A kind of method that active acid is extracted from japanese ardisia root - Google Patents
A kind of method that active acid is extracted from japanese ardisia root Download PDFInfo
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- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/30—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having two double bonds between ring members or between ring members and non-ring members
- C07D207/32—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
- C07D207/33—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms with substituted hydrocarbon radicals, directly attached to ring carbon atoms
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Abstract
The invention provides a kind of method that active acid is extracted from japanese ardisia root, methods described comprises the following steps：S1：Prepare japanese ardisia root powder；S2：Prepare multicomponent compound extracted solution；S3：The preparation of crude product；S4：Crude product refining；S5：Purifies and separates.Methods described is by various extraction processes, parameter, the comprehensive selection of key element and combination, so that achieving best synergy between each key element, purpose product can be obtained with high yield and high-purity, so that raw material availability has reached maximization, industrially especially active components of plants extractive technique field has good application potential and industrial prospect.
Description
Technical field
The present invention relates to a kind of method that active component is extracted from plant, relate more specifically to a kind of from japanese ardisia root
A kind of method of active acid is extracted, belongs to active components of plants extractive technique field.
Background technology
In field of plant extraction, extracted during active component has become current drug field from plant and find active component
A kind of important method, such as common active acid in plant, such as pyrroles acid derivative are exactly a kind of common active acid, and 4-
(2- formoxyl -5- hydroxymethylpyrrol -1- bases) butyric acid is exactly a kind of pyrroles's acid substance with multi-medicament active function, its tool
There is multiple biological activities breast protection liver etc., its chemical structural formula is as follows：
Just because of such bioactivity of the material, people have carried out substantial amounts of further investigation to its extracting method, and
Certain achievement in research is achieved, it is specific as follows：
Outstanding person et al. in relative (" the assay research of pyrroles's butyric acid in radix ranunculi ternati ", Chinese Pharmaceutical Journal, 2012,47 (2),
A kind of method that above-mentioned PA is extracted in 101-103) disclosing radix ranunculi ternati from cohosh.
Chin Young-Won et al. (" Hepatoprotective Pyrrole Derivatives of Lycium
Chinese Fruits ", Bioorganic＆Medicinal Chemistry Letters, 2003,13,79-81) disclose one
The method that above-mentioned pyrroles's acid is extracted from matrimony vine of solanaceae plant is planted, by complicated PROCESS FOR TREATMENT, wherein one is identified
It is exactly above-mentioned PA to plant material.
Chinese patent application CN103073479A reports the side that azole derivatives are separated from the overgrown with weeds green grass or young crops of crucifer
Method, has obtained above-mentioned pyrroles's acid compound of certain content.
As described above, the method for having been disclosed for various extraction active acids in the prior art, but carrying for its novelty
Take and separation method, still suffer from exigence, this is also the study hotspot in the current field.
The Japanese ardisia, small blue or green, the short tea of alias, is Myrsinacea, Ardisa undershrub or semishrub plant, Japanese ardisia complete stool
And root hyoscine, control pulmonary tuberculosis, spitting of blood, cough, chronic bronchitis effect very well, it is the conventional Chinese herbal medicine of folks of china.However,
But not yet find that method or technique that pyrroles's acid is extracted from the Japanese ardisia are seen in report in the prior art, it belongs to pyrroles
Acid extracts the research blank in field.
The present inventor is directed to above-mentioned latent defect, by the research for many years being distributed to plant variety and effective component, and opens
A kind of new extraction process of above-mentioned 4- (2- formoxyl -5- hydroxymethylpyrrol -1- bases) butyric acid is sent out, it is realized from purple first
Extracting and developing goes out the Particular craft of pyrroles's acrylic component in Taurus section plant japanese ardisia root, there is provided the new plant of such material
Thing is originated, and fully meets the common requirements of medicine and pharmaceutical arts to pyrroles's acid, with great commercial introduction and
Market application foreground.
The content of the invention
As described above, the extraction in order to widen pyrroles's acids active material is originated, novel separation method, the present invention are developed
People by after substantial amounts of creative work, have developed it is a kind of from japanese ardisia root extract PA new method, so as to complete
The present invention.
More specifically, active acid (i.e. 4- (2- formoxyl -5- hydroxyls are extracted from japanese ardisia root the invention provides one kind
Methylpyrrole -1- bases) butyric acid) method, methods described comprises the following steps：
S1：Prepare japanese ardisia root powder；
S2：Prepare multicomponent compound extracted solution；
S3：The preparation of crude product；
S4：Crude product refining；
S5：Purifies and separates.
In methods described of the invention, the step S1 is specific as follows：
Take mass water content and dry japanese ardisia root less than 2%, use NaHCO3Aqueous solution soaking 20-30 hours, Ran Houyong
Deionized water is washed 2-3 times, after drying completely, is crushed, 100 mesh sieves is crossed, so as to obtain japanese ardisia root powder (i.e. raw material).
Wherein, in this step, the NaHCO3The mass percent concentration of the aqueous solution is 1-3%, for example can for 1%,
2% or 3%.
Wherein, in this step, NaHCO is used3Aqueous solution soaking 20-30 hours, can for example soak 20 hours, 25 hours or
30 hours.
In methods described of the invention, the step S2 is specific as follows：
By acetone, petroleum ether and the cis- 3- hexenes ester of 2-Methyl Butyric Acid according to mass ratio 2-3:1.5-2.5:1 ratio is mixed
Close, obtain mixed liquor；Extraction aid is subsequently adding, stirring mixing 5-10 minutes, obtains final product multicomponent compound extracted solution at room temperature.
Wherein, in this step, the mass ratio of acetone, petroleum ether and the cis- 3- hexenes ester of 2-Methyl Butyric Acid is 2-3:1.5-
2.5:1, " 2-3 " therein for example can be 2,2.5 or 3, and " 1.5-2.5 " therein for example can be 1.5,2 or 2.5, this three's
Mass ratio can be any ratio being mutually combined out of any specific point value in these scopes.
Wherein, in this step, the extraction aid is 1- butyl -3- methylimidazole toluenesulfonates (CAS：
410522-18-8), 1- butyl -3- methylimidazole trifluoroacetates (CAS：174899-94-6) or 1- butyl -3- methylimidazoles
Dicyan amine salt (CAS：Any one in 448245-52-1), preferably 1- butyl -3- methylimidazoles dicyan amine salt.
Wherein, in this step, the consumption of the extraction aid is the mixed liquor (i.e. acetone, petroleum ether and 2- methyl fourth
The mixed liquor of the cis- 3- hexenes ester of acid) weight 5-15%, preferably 8-12%, most preferably 10%.
In methods described of the invention, the step S3 is specific as follows：
The japanese ardisia root powder of step S1 is added in the multicomponent compound extracted solution of step S2, and is warming up to 50-60
DEG C, to stir 60-90 minutes, filtering obtains filtrate and filter residue；Filter residue is repeated into operation 2-3 times；Merge all of filter
Liquid, mixes with isometric deionized water, fully vibration, isolates organic layer, is concentrated under reduced pressure, and obtains paste, as crude product.
Wherein, in this step, the raw material and the mass ratio of multicomponent compound extracted solution are 1:6-10, for example, can be 1:
6、1:7、1:8、1:9 or 1:10.
Wherein, in this step, " filter residue is repeated into operation 2-3 times " and to be meant rejoin filter residue multigroup
In point compound extracted solution, and it is warming up to 50-60 DEG C, stirs 60-90 minute, filter, filtrate and filter residue is obtained again；And filter every time
Slag is also 1 with the mass ratio of multicomponent compound extracted solution:6-10, for example, can be 1:6、1:7、1:8、1:9 or 1:10.
In methods described of the invention, the step S4 is specific as follows：
The crude product that step S3 is obtained is added to saturation NaHCO3In the aqueous solution, vibration makes it fully dissolve, and then takes out
Filter, obtains filtrate and insoluble matter；By the acetone solution that insoluble matter temperature is 40-50 DEG C, so as to obtain acetone soln；Will be above-mentioned
Filtrate and acetone soln mixing, abundant vibration, standing, take upper organic phase, are concentrated under reduced pressure, and obtain medicinal extract.
Wherein, in this step, the crude product and saturation NaHCO3The mass volume ratio of the aqueous solution is 1:10-15g/ml,
10-15ml saturations NaHCO will be added to per 1g crude products3In the aqueous solution.
Wherein, in this step, insoluble matter and the mass volume ratio of acetone are 1:4-8g/ml, i.e., use per 1g insoluble matters
4-8ml acetone is dissolved.
In methods described of the invention, the step S5 is specific as follows：
The medicinal extract of step S4 is dissolved in ethyl acetate, is subsequently poured into chromatographic column, the chromatographic column is with volume basis
Than meter, 30% neutral silica gel, 20% acidic silica gel and 30% neutral alumina are filled with successively from top to bottom；Second is used successively
Ether, n-hexane, mass percent concentration are that 10% n-butanol aqueous solution is eluted, and collect n-butanol elution fraction, decompression
Concentration, obtains final product the active acid (i.e. 4- (2- formoxyl -5- hydroxymethylpyrrol -1- bases) butyric acid).
Wherein, in this step, the medicinal extract and chromatographic column filler (neutral silica gel i.e. therein, acidic silica gel and neutrality
Aluminum oxide) mass ratio be 1:100-150, for example, can be 1:100、1:120：、1:140 or 1:150.
Wherein, in this step, medicinal extract is dissolved in ethyl acetate, the amount of the ethyl acetate can relatively soak just
Cream dissolving is advisable completely, can so facilitate follow-up elution action.
Wherein, in this step, the numerical value of the neutral silica gel, acidic silica gel and neutral alumina refers to that packing volume is accounted for
The ratio of chromatographic column cumulative volume, such as by taking 30% neutral silica gel as an example, its packing volume is the 30% of chromatogram column volume, the acid
Property silica gel and neutral alumina also have same implication.
Wherein, in this step, the neutral silica gel and neutral alumina are very known commercially available prod, for example its
Granularity can be 200-300 mesh, and this is no longer going to repeat them.
Wherein, in this step, the acidic silica gel is obtained in accordance with the following steps：
I, the neutral silica gel of the mesh of 200 mesh -300 is baked 3-5 hours in 500 DEG C of Muffle furnace, after baking end, cooling
To 250-280 DEG C and 40-50 minutes is incubated, is finally cooled to room temperature, obtain activated silica gel；
II, in activated silica gel add be added dropwise add mass percent concentration for 20-30% aqueous sulfuric acid, the work
SiClx glue is 1 with the mass ratio of institute's sulfur acid in aqueous sulfuric acid:0.02-0.06, then abundant shaken well, has been vacuum dried
Entirely, acidic silica gel is obtained final product.
Wherein, in this step, ether, n-hexane, the n-butanol water that mass percent concentration is 10% used is eluted
The volume of solution is respectively the 2-3 of chromatographic column packing course volume (i.e. the cumulative volume of neutral silica gel, acidic silica gel and neutral alumina)
Again, 2-2.5 times and 0.6-1.2 times.
As described above, the invention provides a kind of completely new approach that active acid is extracted from japanese ardisia root, methods described is led to
The selection of multiple key elements is crossed, it is such as suitable to extract operation, the preparation of extract solution, the selection of eluent, the filling species of chromatographic column
Etc. and achieve good extraction effect, the extraction source of pyrroles's acid has been opened up, and by experimental exploring come to work
The optimised process that skill condition is optimized and is finally obtained extraction is combined, with wide industrial application value.
Specific embodiment
Below by specific embodiment, the present invention is described in detail, but the purposes of these exemplary embodiments and
Purpose is only used for enumerating the present invention, not constitutes any type of any restriction to real protection scope of the invention, more non-to incite somebody to action
Protection scope of the present invention is confined to this.
Preparation example：The preparation of acidic silica gel
I, the neutral silica gel of 200-300 mesh is baked in 500 DEG C of Muffle furnace 4 hours, after baking end, be cooled to
260 DEG C and be incubated 45 minutes, be finally cooled to room temperature, obtain activated silica gel；
II, in activated silica gel add be added dropwise add mass percent concentration be 25% aqueous sulfuric acid, the activation
Silica gel is 1 with the mass ratio of institute's sulfur acid in aqueous sulfuric acid:0.04, then abundant shaken well, is vacuum dried completely, obtains final product
Acidic silica gel.
Unless otherwise indicated, the acidic silica gel for using in the following example is the acidic silica gel that the preparation example is obtained.
Embodiment 1
S1：Prepare japanese ardisia root powder
Take mass water content and dry japanese ardisia root less than 2%, with the NaHCO that mass percent concentration is 1%3The aqueous solution
Immersion 20 hours, is then washed with deionized 2-3 times, after drying completely, crushes, 100 mesh sieves is crossed, so as to obtain japanese ardisia root
Powder (i.e. raw material)；
S2：Prepare multicomponent compound extracted solution
By acetone, petroleum ether and the cis- 3- hexenes ester of 2-Methyl Butyric Acid according to mass ratio 2:1.5:1 ratio mixing, obtains
Mixed liquor；The extraction aid 1- butyl -3- methylimidazole dicyan amine salt for mixed liquor weight 5% are subsequently adding, stir mixed at room temperature
Close 5 minutes, obtain final product multicomponent compound extracted solution；
S3：The preparation of crude product
During the japanese ardisia root powder of step S1 is added into the multicomponent compound extracted solution of step S2 (wherein described raw material with
The mass ratio of multicomponent compound extracted solution is 1:6), and it is warming up to 50 DEG C, stirs 90 minutes, filtering obtains filtrate and filter residue；Will
Filter residue repeats the operation 2 times；Merge all of filtrate, mix with isometric deionized water, fully vibration, isolated
Machine layer, is concentrated under reduced pressure, and obtains paste, as crude product；
S4：Crude product refining
The crude product that step S3 is obtained is added to saturation NaHCO3(crude product and saturation saturation NaHCO in the aqueous solution3Water
The mass volume ratio of solution is 1:10g/ml), vibrate, it is fully dissolved, then suction filtration, obtain filtrate and insoluble matter；Will not
Molten thing temperature is that (mass volume ratio of the insoluble matter and acetone is 1 for 40 DEG C of acetone solution:4g/ml), so as to obtain acetone
Solution；Above-mentioned filtrate and acetone soln are mixed, fully vibrated, stood, take upper organic phase, be concentrated under reduced pressure, obtain medicinal extract；
S5：Purifies and separates
The medicinal extract of step S4 is dissolved in appropriate ethyl acetate, is subsequently poured into chromatographic column, the chromatographic column is with volume
Percentages, are filled with 30% neutral silica gel, 20% acidic silica gel and the 30% neutral alumina (medicinal extract successively from top to bottom
It is 1 with the mass ratio of chromatographic column filler:100)；2 times of ether, 2 times of chromatographic column packing course bodies of chromatographic column packing course volume are used successively
Long-pending n-hexane, the n-butanol aqueous solution that the mass percent concentration of 0.6 times of chromatographic column packing course volume is 10% are eluted, and are received
Collection n-butanol elution fraction, is concentrated under reduced pressure, and obtains final product the active acid (i.e. 4- (2- formoxyl -5- hydroxymethylpyrrol -1- bases) fourth
Acid), its purity is determined as 99.3% by HPLC, and its nuclear-magnetism characterize data is as follows：
1H NMR(CD3OD,600MHz)δ：9.42 (1H, s, H-6), 6.96 (1H, d, J=4.2Hz, H-3), 6.27 (1H,
D, J=4.2Hz, H-4), 4.57 (2H, s, H-7), 4.33 (2H, t, J=7.8Hz, H-4 '), 2.17 (2H, t, J=7.2Hz, H-
2 '), 1.94 (2H, dt, J=7.8,7.2Hz, H-3 ').
13C NMR(CD3OD,150MHz)δ：181.2(s,C-1′),180.1(d,C-6),144.6(s,C-5),133.4
(s,C-2),126.2(d,C-3),111.5(d,C-4),56.3(t,C-7),46.5(t,C-4′),35.6(t,C-2′),29.1
(t,C-3′)。
By calculating, in the present embodiment, often processing the japanese ardisia root powder in 100g steps S1, to obtain 94.3mg above-mentioned
Active acid.
Embodiment 2
S1：Prepare japanese ardisia root powder
Take mass water content and dry japanese ardisia root less than 2%, with the NaHCO that mass percent concentration is 2%3The aqueous solution
Immersion 25 hours, is then washed with deionized 2-3 times, after drying completely, crushes, 100 mesh sieves is crossed, so as to obtain japanese ardisia root
Powder (i.e. raw material)；
S2：Prepare multicomponent compound extracted solution
By acetone, petroleum ether and the cis- 3- hexenes ester of 2-Methyl Butyric Acid according to mass ratio 2.5:2:1 ratio mixing, obtains
Mixed liquor；The extraction aid 1- butyl -3- methylimidazole dicyan amine salt for mixed liquor weight 10% are subsequently adding, stir mixed at room temperature
Close 8 minutes, obtain final product multicomponent compound extracted solution；
S3：The preparation of crude product
During the japanese ardisia root powder of step S1 is added into the multicomponent compound extracted solution of step S2 (wherein described raw material with
The mass ratio of multicomponent compound extracted solution is 1:8), and it is warming up to 55 DEG C, stirs 75 minutes, filtering obtains filtrate and filter residue；Will
Filter residue repeats the operation 3 times；Merge all of filtrate, mix with isometric deionized water, fully vibration, isolated
Machine layer, is concentrated under reduced pressure, and obtains paste, as crude product；
S4：Crude product refining
The crude product that step S3 is obtained is added to saturation NaHCO3(crude product and saturation saturation NaHCO in the aqueous solution3Water
The mass volume ratio of solution is 1:15g/ml), vibrate, it is fully dissolved, then suction filtration, obtain filtrate and insoluble matter；Will not
Molten thing temperature is that (mass volume ratio of the insoluble matter and acetone is 1 for 50 DEG C of acetone solution:8g/ml), so as to obtain acetone
Solution；Above-mentioned filtrate and acetone soln are mixed, fully vibrated, stood, take upper organic phase, be concentrated under reduced pressure, obtain medicinal extract；
S5：Purifies and separates
The medicinal extract of step S4 is dissolved in appropriate ethyl acetate, is subsequently poured into chromatographic column, the chromatographic column is with volume
Percentages, are filled with 30% neutral silica gel, 20% acidic silica gel and the 30% neutral alumina (medicinal extract successively from top to bottom
It is 1 with the mass ratio of chromatographic column filler:120)；3 times of ether, 2.2 times of chromatographic column packing courses of chromatographic column packing course volume are used successively
The n-hexane of volume, the n-butanol aqueous solution that the mass percent concentration of 1.2 times of chromatographic column packing course volumes is 10% are eluted,
N-butanol elution fraction is collected, is concentrated under reduced pressure, obtain final product the active acid (i.e. 4- (2- formoxyl -5- hydroxymethylpyrrol -1- bases) fourth
Acid), its purity is determined as 98.9% by HPLC, and its nuclear-magnetism characterize data is with embodiment 1.
By calculating, in the present embodiment, often processing the japanese ardisia root powder in 100g steps S1, to obtain 94.6mg above-mentioned
Active acid.
Embodiment 3
S1：Prepare japanese ardisia root powder
Take mass water content and dry japanese ardisia root less than 2%, with the NaHCO that mass percent concentration is 3%3The aqueous solution
Immersion 30 hours, is then washed with deionized 2-3 times, after drying completely, crushes, 100 mesh sieves is crossed, so as to obtain japanese ardisia root
Powder (i.e. raw material)；
S2：Prepare multicomponent compound extracted solution
By acetone, petroleum ether and the cis- 3- hexenes ester of 2-Methyl Butyric Acid according to mass ratio 3:2.5:1 ratio mixing, obtains
Mixed liquor；The extraction aid 1- butyl -3- methylimidazole dicyan amine salt for mixed liquor weight 15% are subsequently adding, stir mixed at room temperature
Close 10 minutes, obtain final product multicomponent compound extracted solution；
S3：The preparation of crude product
During the japanese ardisia root powder of step S1 is added into the multicomponent compound extracted solution of step S2 (wherein described raw material with
The mass ratio of multicomponent compound extracted solution is 1:10), and it is warming up to 60 DEG C, stirs 60 minutes, filtering obtains filtrate and filter residue；
Filter residue is repeated into the operation 3 times；Merge all of filtrate, mix with isometric deionized water, fully vibration, isolate
Organic layer, is concentrated under reduced pressure, and obtains paste, as crude product；
S4：Crude product refining
The crude product that step S3 is obtained is added to saturation NaHCO3(crude product and saturation saturation NaHCO in the aqueous solution3Water
The mass volume ratio of solution is 1:12g/ml), vibrate, it is fully dissolved, then suction filtration, obtain filtrate and insoluble matter；Will not
Molten thing temperature is that (mass volume ratio of the insoluble matter and acetone is 1 for 45 DEG C of acetone solution:6g/ml), so as to obtain acetone
Solution；Above-mentioned filtrate and acetone soln are mixed, fully vibrated, stood, take upper organic phase, be concentrated under reduced pressure, obtain medicinal extract；
S5：Purifies and separates
The medicinal extract of step S4 is dissolved in appropriate ethyl acetate, is subsequently poured into chromatographic column, the chromatographic column is with volume
Percentages, are filled with 30% neutral silica gel, 20% acidic silica gel and the 30% neutral alumina (medicinal extract successively from top to bottom
It is 1 with the mass ratio of chromatographic column filler:150)；Filled out using 2.5 times of ether of chromatographic column packing course volume, 2.5 times of chromatographic columns successively
The n-hexane of layer volume, the n-butanol aqueous solution that the mass percent concentration of 0.8 times of chromatographic column packing course volume is 10% are washed
It is de-, n-butanol elution fraction is collected, it is concentrated under reduced pressure, obtain final product the active acid (i.e. 4- (2- formoxyl -5- hydroxymethylpyrrols -1-
Base) butyric acid), its purity is determined as 98.8% by HPLC, and its nuclear-magnetism characterize data is with embodiment 1.
By calculating, in the present embodiment, often processing the japanese ardisia root powder in 100g steps S1, to obtain 94.1mg above-mentioned
Active acid.
Embodiment 4-9
Embodiment 4-6：Except the extraction aid in embodiment 1-3 is replaced with into 1- butyl -3- methylimidazoles to methylbenzene respectively
Outside sulfonate, other all operations are constant, so as to sequentially obtain embodiment 4-6.
Embodiment 7-9：Except the extraction aid in embodiment 1-3 is replaced with into 1- butyl -3- methylimidazole trifluoroacetic acids respectively
Outside salt, other all operations are constant, so as to sequentially obtain embodiment 7-9.
For the japanese ardisia root powder in every 100g steps S1, the yield and purity of gained active acid see the table below 1：
Table 1：The investigation of extraction aid
As can be seen here, the species of extraction aid has significant impact for final product recovery rate and purity, wherein, 1-
There is butyl -3- methylimidazole dicyan amine salt best helping to put forward effect, and 1- butyl -3- methylimidazoles toluenesulfonate and
The recovery rate and purity all decreases to some degree, especially recovery rate of 1- butyl -3- methylimidazole trifluoroacetates.
Embodiment 10-12
Except respectively by the NaHCO in embodiment 1-3 steps S13Aqueous solution soaking treatment dispenses outer, other all operations
It is constant, so as to sequentially obtain embodiment 10-12.
For the japanese ardisia root powder in every 100g steps S1, the yield and purity of gained active acid see the table below 2：
Table 2：NaHCO3The influence of aqueous solution soaking treatment
As can be seen here, as unused NaHCO3When the aqueous solution carries out immersion treatment, active acid total amount and purity decrease,
It is likely to be and has extracted the impurity of some effects subsequent extracted by the immersion, so as to improves yield to a certain extent
And purity, inventor is intended to further be studied.
Embodiment 13-15
Except the cis- 3- hexenes ester of 2-Methyl Butyric Acid in multicomponent compound extracted solution in embodiment 1-3 steps S2 is given respectively
To dispense outer (only use the mixture of acetone and petroleum ether as compound extracted solution), other are all operate it is constant, from
And sequentially obtained embodiment 13-15.
For the japanese ardisia root powder in every 100g steps S1, the yield and purity of gained active acid see the table below 3：
Table 3：The investigation of multicomponent compound extracted solution
As can be seen here, in the multicomponent compound extracted solution of the invention, the cis- 3- hexenes ester of 2-Methyl Butyric Acid must
Few, after the component is dispensed, although the purity of gained active acid does not change substantially, but yield has significantly
Reduce.
Embodiment 16-24
Embodiment 16-18：Chromatographic column in the step of except embodiment 1-3 S5 only loads (loadings with neutral silica gel respectively
Still it is the overall accumulated amount of original three kinds of fillers) outward, other operations are constant, so as to sequentially obtain embodiment 16-18.
Embodiment 19-21：Chromatographic column in the step of except embodiment 1-3 S5 only loads (loadings with acidic silica gel respectively
Still it is the overall accumulated amount of original three kinds of fillers) outward, other operations are constant, so as to sequentially obtain embodiment 19-21.
Embodiment 22-24：Chromatographic column in the step of except embodiment 1-3 S5 only loads (filling with neutral alumina respectively
Amount is still the overall accumulated amount of original three kinds of fillers) outward, other operations are constant, so as to sequentially obtain embodiment 22-24.
For the japanese ardisia root powder in every 100g steps S1, the yield and purity of gained active acid see the table below 4：
Table 4：The investigation of chromatographic column filler
As can be seen here, in purification procedures of the invention, the Fillers selection of chromatographic column is extremely important, only makes simultaneously
With the combination of neutral silica gel, acidic silica gel and neutral alumina, good extraction effect of the invention and purity could be obtained, and adopted
During with single filler, causing yield and purity to have significantly reduces.
Embodiment 25-36
Embodiment 25-27：A use quality percent concentration is 10% positive fourth respectively in the step of except embodiment 1-3 S5
Alcohol solution is eluted (ether and the n-hexane wash-out before eliminating), and outward, other operations are constant, so that sequentially
Embodiment 25-27 is arrived.
Embodiment 28-30：Only the use of ether and mass percent concentration is respectively 10% in the step of except embodiment 1-3 S5
N-butanol aqueous solution eluted (n-hexane wash-out) before eliminating outward, other operations be constant, so as to sequentially must
Embodiment 28-30 is arrived.
Embodiment 31-33：It is using n-hexane and mass percent concentration only respectively in the step of except embodiment 1-3 S5
10% n-butanol aqueous solution is eluted (the ether wash-out before eliminating), and outward, other operations are constant, so that sequentially
Embodiment 31-33 is obtained.
Embodiment 34-36：Only eluted using ether and n-hexane respectively in the step of except embodiment 1-3 S5 and (saved
Omit last n-butanol aqueous solution wash-out) and collect ether and n-hexane eluent and carry out outside subsequent purification, other operations
It is constant, so as to sequentially obtain embodiment 34-36.
For the japanese ardisia root powder in every 100g steps S1, the yield and purity of gained active acid see the table below 5：
Table 5：The investigation of eluant, eluent
As can be seen here, in purification procedures of the invention, the conjunctive use of eluant, eluent, successively elute it is extremely important：
When the n-butanol aqueous solution that only use quality percent concentration is 10% is eluted, although yield increases, but purity
It is greatly lowered；When being eluted using ether and the n-butanol aqueous solution and entered using n-hexane and the n-butanol aqueous solution
During row wash-out, either yield or purity are significantly reduced；And when dispensing n-butanol aqueous solution and eluting, then product
Yield and purity reduction are the most notable, without the necessity of any industrializing implementation.All these is demonstrated simultaneously
The importance and unobviousness sequentially eluted using ether, n-hexane and the n-butanol aqueous solution, and achieve meaning
Unimaginable technique effect.
As described above, the invention provides a kind of method that active acid is extracted from japanese ardisia root, methods described is by more
Extraction process, parameter, the comprehensive selection of key element and combination are planted, so that achieving best collaboration effect between each key element
Really, purpose product can be obtained with high yield and high-purity so that raw material availability has reached maximization, industrially especially
It is that active components of plants extractive technique field has good application potential and industrial prospect.
It should be appreciated that the purposes of these embodiments is merely to illustrate the present invention and is not intended to limit protection model of the invention
Enclose.Additionally, it will also be appreciated that after technology contents of the invention have been read, those skilled in the art can make each to the present invention
Plant and change, change and/or modification, all these equivalent form of value equally falls within the guarantor that the application appended claims are limited
Within the scope of shield.
Claims (4)
1. the method that one kind extracts active acid 4- (2- formoxyl -5- hydroxymethylpyrrol -1- bases) butyric acid from japanese ardisia root, it is described
Method comprises the following steps：
S1：Prepare japanese ardisia root powder；
S2：Prepare multicomponent compound extracted solution；
S3：The preparation of crude product；
S4：Crude product refining；
S5：Purifies and separates；
The step S1 is specific as follows：
Take mass water content and dry japanese ardisia root less than 2%, with NaHCO3 aqueous solution soakings 20-30 hours, then spend from
Sub- water washing 2-3 times, after drying completely, crushes, 100 mesh sieves is crossed, so as to obtain japanese ardisia root powder；
The step S2 is specific as follows：
By acetone, petroleum ether and the cis- 3- hexenes ester of 2-Methyl Butyric Acid according to mass ratio 2-3:1.5-2.5:1 ratio mixing, obtains
To mixed liquor；Extraction aid is subsequently adding, stirring mixing 5-10 minutes, obtains final product multicomponent compound extracted solution at room temperature；
The extraction aid be 1- butyl -3- methylimidazoles toluenesulfonate, 1- butyl -3- methylimidazoles trifluoroacetate or
Any one in 1- butyl -3- methylimidazole dicyan amine salt；
The step S3 is specific as follows：
The japanese ardisia root powder of step S1 is added in the multicomponent compound extracted solution of step S2, and is warming up to 50-60 DEG C, stirred
Mix 60-90 minutes, filter, obtain filtrate and filter residue；Filter residue is repeated into operation 2-3 times；Merge all of filtrate, with etc.
Organic layer is isolated in the deionized water mixing of volume, fully vibration, is concentrated under reduced pressure, and obtains paste, as crude product；
The step S4 is specific as follows：
The crude product that step S3 is obtained is added in the saturation NaHCO3 aqueous solution, is vibrated, it is fully dissolved, then suction filtration, obtained
To filtrate and insoluble matter；By the acetone solution that insoluble matter temperature is 40-50 DEG C, so as to obtain acetone soln；By above-mentioned filtrate
With acetone soln mixing, fully vibration, standing, upper organic phase is taken, be concentrated under reduced pressure, obtain medicinal extract；
The step S5 is specific as follows：
The medicinal extract of step S4 is dissolved in ethyl acetate, is subsequently poured into chromatographic column, the chromatographic column with volume percentage,
It is filled with 30% neutral silica gel, 20% acidic silica gel and 30% neutral alumina successively from top to bottom；Ether, just oneself are used successively
Alkane, mass percent concentration are that 10% n-butanol aqueous solution is eluted, and collect n-butanol elution fraction, are concentrated under reduced pressure, i.e.,
Obtain the active acid；
The acidic silica gel is obtained in accordance with the following steps：
I, the neutral silica gel of the mesh of 200 mesh -300 is baked 3-5 hours in 500 DEG C of Muffle furnace, after baking end, be cooled to
250-280 DEG C and 40-50 minutes is incubated, is finally cooled to room temperature, obtain activated silica gel；
II, it is the aqueous sulfuric acid of 20-30%, the activated silica gel and sulfuric acid to mass percent concentration is added in activated silica gel
The mass ratio of institute's sulfur acid is 1 in the aqueous solution:0.02-0.06, then abundant shaken well, is vacuum dried completely, obtains final product acidity
Silica gel.
2. method according to claim 1, it is characterised in that：The extraction aid is 1- butyl -3- methylimidazole cdicynanmides
Salt.
3. method according to claim 1, it is characterised in that：The quality of the medicinal extract and chromatographic column filler in step S5
Than being 1:100-150.
4. the method according to claim any one of 1-3, it is characterised in that：Wash-out ether, n-hexane, quality hundred used
Divide the volume respectively chromatographic column packing course volume of the n-butanol aqueous solution that specific concentration is 10% 2-3 times, 2-2.5 times and 0.6-
1.2 times.
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Denomination of invention: A method of extracting active acid from Ardisia root Effective date of registration: 20201103 Granted publication date: 20170630 Pledgee: China Construction Bank Corporation Taizhou pharmaceutical high tech Zone sub branch Pledgor: TAIZHOU DANDING BIOLOGICAL TECHNOLOGY Co.,Ltd. Registration number: Y2020980007484 |