CN105175426B - A kind of method of the extraction purification Bergenin from treebine stem - Google Patents
A kind of method of the extraction purification Bergenin from treebine stem Download PDFInfo
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- CN105175426B CN105175426B CN201510657480.3A CN201510657480A CN105175426B CN 105175426 B CN105175426 B CN 105175426B CN 201510657480 A CN201510657480 A CN 201510657480A CN 105175426 B CN105175426 B CN 105175426B
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- bergenin
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- macroreticular resin
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- treebine stem
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D493/00—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system
- C07D493/02—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains two hetero rings
- C07D493/04—Ortho-condensed systems
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/13—Crystalline forms, e.g. polymorphs
Abstract
The invention discloses a kind of methods of the extraction purification Bergenin from treebine stem, and treebine stem is crushed, sieving, are 1 according to mass volume ratio：8 ~ 20 add in ethanol solution refluxing extraction, separation of solid and liquid obtains filtrate, filtrate is concentrated, macroreticular resin Static Adsorption is added in into concentrate, by the macroreticular resin upper prop after absorption, impurity, elution first are removed with distilled water, ethanol solution successively, collects eluent, eluent is concentrated to give Bergenin crude product, Bergenin crude product is obtained into Bergenin sterling with ethyl alcohol recrystallization.Raw material sources of the present invention are extensive, cheap, and production cost is low, and working condition is mild, and environmental pollution is small.
Description
Technical field
The present invention relates to a kind of methods extracted from treebine stem, purify Bergenin, belong to natural drug utilization
Technical field.
Background technology
Treebine stem（Cissus Pteroclada Hayata）Belong to Vitaceae（Vitaceae）Cissus（Cissus Linn.）Plant, gains the name also known as square rattan, Herba Lycopodii, leech rattan etc. due to its stem, branch, petiole are in sharp square.Treebine stem is main
The southern area such as the Guangxi, Guangdong, Yunnan in China is distributed in, resourceful, property is sour, puckery, sweet, flat, has promoting blood circulation to remove blood stasis, dispels
Rheumatism, relaxing muscles and tendons network, anti-inflammatory and other effects, in China, among the people to be widely used in arthralgia pain due to rheumatism, lumbar muscle strain, wound, bleeding etc. ailing
Treatment has good efficacy.Such as Bose Region peasant directly mashes external application with treebine stem fresh leaf and stops blooding；Baise hospital is by four directions
Pulvis and injection is made in rattan, for treating the diseases such as interior traumatic hemorrhage, duodenal hemorrhage, gastritis.Treebine stem record in《Extensively
Western Chinese herbal medicine》、《Medicinal Plants of Guangxi register》、《Chinese Pharmacopoeia》（Version one in 1977）、《Dictionary of medicinal plant》It waits in ancient books and records, because
Its abundance, cheap, curative effect is extensive and notable, is a kind of national medicinal plant of characteristic for having value of exploiting and utilizing
Object.Although treebine stem is among the people with long medicinal history and experience in Chinese Minority Nationalities, due to treebine stem chemical composition
With the shortage of pharmacological research, in the utilization of treebine stem active ingredient, there has been no the appearance of depths products, on the market
It is sold still in the form of crude drug, it is cheap.The structure of Bergenin：
Bergenin also known as brave youngster's grass element, arolisic acid B, molecular formula C14H16O9, it is that one kind has isocoumarin structure
Native compound.Bergenin and its derivative have a variety of pharmacological activity such as anti-inflammatory, cough-relieving, analgesia, antibacterial, anti-oxidant.
Bergenin be used as antisussive and expectorant agent record in《Pharmacopoeia of People's Republic of China》In each version, bronchus is used clinically for
The treatment of the diseases such as inflammation, pulmonary emphysema, gastritis has good efficacy, is that a kind of pharmacological activity is extensive, evident in efficacy, toxic side effect
Low natural drug.
Due to the difficulty of chemical synthesis, Bergenin is mainly with extraction, method of purification preparation from natural medicinal plant.Mesh
Before, the report of Bergenin extracting method is more, and less for the report of its purification process.The extracting method master of Bergenin
There are soxhlet extraction method, ultrasonic extraction, super critical extraction, organic solvent reflux extraction, alkali extraction method etc..However,
Method currently used for purifying Bergenin extract is difficult to prepare with scale.If silica gel column chromatography is common point of laboratory
From means, but silica gel cannot recycle, and eluant, eluent is the toxic organic solvents such as petroleum ether, ethyl acetate, chloroform, of high cost, operation
Difficulty is big, it is impossible to be used in prepared by industrialization；Recrystallization method, it is necessary to be that the content of Bergenin in solution to be crystallized is up to certain journey
It could be used when spending, common Bergenin extract, before no enrichment by other methods, compare and be difficult to crystallize and nothing
Method is directly purified with recrystallization method.In consideration of it, current organic solvent extraction, which still rests on acquisition, contains Bergenin crude extract
Stage, and Bergenin sterling is prepared on a large scale using the extraction method, it is necessary to which developing can industrialization amplification, at low cost
Purification process.
The content of the invention
It is an object of the invention to provide a kind of suitable industrialization to amplify, at low cost that bulk pharmaceutical chemicals rock is prepared from treebine stem
The method of Chinese cabbage element.
What the present invention was achieved through the following technical solutions.
A kind of method of the extraction purification Bergenin from treebine stem, includes the following steps：
（1）Treebine stem is crushed, is sieved, is 1 according to mass volume ratio：8 ~ 20 ethanol solution reflux for adding in 40 ~ 60% carry
It takes；
（2）Separation of solid and liquid obtains filtrate, concentrates the filtrate to the 1/2 ~ 1/3 of its original volume, while recycling design;
（3）When into concentrate, addition macroreticular resin Static Adsorption 6 ~ 8 is small, macroreticular resin is selected from AB-8, D3520, D101；
（4）By the macroreticular resin upper prop after absorption, first impurity is removed with distilled water, 10% ethanol solution successively；
（5）It is eluted with 20% ethanol solution, collects 20% ethanol eluate；
（6）20% ethanol eluate is concentrated, obtains Bergenin crude product；
（7）Bergenin crude product is obtained into Bergenin sterling with ethyl alcohol recrystallization.
Above-mentioned steps（2）In, the method for separation of solid and liquid for filtering or centrifuges, and filtrate is concentrated into Bergenin concentration 2
mg/mL。
Above-mentioned steps（4）In, remove impurity with the distilled water of 2 times of column volumes and 10% ethyl alcohol of 2 times of column volumes.
Above-mentioned steps（5）In, with 20% ethanol elution of 7 times of column volumes.
Above-mentioned steps（6）In, the method for concentration is vacuum concentration or film concentration.
Above-mentioned steps（7）In, it is recrystallized with 95% ethyl alcohol.
The advantage of the invention is that：Using 40 ~ 60% alcoholic extraction, have that green non-poisonous, recovery rate is high, at low cost, easy
The advantages that recycling；Using macroporous resin purification and enrichment, have large amount of adsorption, it is repeatable utilize, be easy to industrialization amplification,
The advantages that at low cost, after macroporous resin purification, can obtain crude product of the purple bergenia herb purity more than 80%；Macroreticular resin is pure
It is Bergenin crude product after change, it is pure as the Bergenin that bulk pharmaceutical chemicals use to obtain satisfaction by ethanol solution recrystallization
Product；Raw material sources of the present invention are extensive, cheap, and production cost is low, and working condition is mild, and environmental pollution is small.
Description of the drawings
Fig. 1 is extraction and purification process flow chart of the present invention；
The static saturated adsorption rate of Fig. 2 difference macroreticular resins；
The static desorption efficiency of Fig. 3 difference macroreticular resins.
Specific embodiment
The invention will be further described by the following examples.
Embodiment 1
By the treebine stem powder of 500 g, 50% ethanol solution is added in, makes the quality of treebine stem powder and the volume ratio of ethyl alcohol
For 1:20, refluxing extraction 1 time, every time 2 h is centrifuged, filtered, discarding waste residue and collect to obtain filtrate, filtrate is concentrated, is recycled simultaneously
Solvent.It is 1 by volume and macroreticular resin quality by the concentrate that concentration is 2.0 mg/mL:1 ratio adds in AB-8 macropore trees
Static Adsorption is carried out in fat, adsorption time is 8 h, then upper prop.First with the 10% of the distilled water of 2 times of column volumes and 2 times of column volumes
Ethyl alcohol removes impurity, is then eluted again with 7 times of 20% ethanol solutions of column volume, collects 20% ethanol elution part.It will elution
Liquid concentrates, and obtains 10.3 g of Bergenin crude product, 8.9 g of Bergenin sterling is obtained after 95% ethyl alcohol recrystallization.
Embodiment 2
By the treebine stem powder of 1Kg, 50% ethanol solution is added in, it is 1 to make the mass volume ratio of treebine stem powder and ethyl alcohol:
15, refluxing extraction 2 times, every time 2 h is centrifuged, filtered, discarding waste residue and collect to obtain filtrate, filtrate is concentrated, while is recycled organic
Solvent.It is 1 by volume and macroreticular resin quality by the concentrate that concentration is 2.0 mg/mL:1 ratio adds in D3520 macropores
Static Adsorption is carried out in resin, adsorption time is 8 h, then upper prop.First with the distilled water of 2 times of column volumes and 2 times of column volumes
10% ethyl alcohol removes impurity.Then eluted again with 7 times of 20% ethanol solutions of column volume, collect 20% ethanol elution part.It will wash
De- liquid concentration, obtains 20.5 g of Bergenin crude product, 17.6 g of Bergenin sterling is obtained after 95% ethyl alcohol recrystallization.
Embodiment 3
By the treebine stem powder of 10 Kg, 60% ethanol solution is added in, the mass volume ratio for making treebine stem powder and ethyl alcohol is
1:10, refluxing extraction 3 times, every time 2 h is centrifuged, filtered, discarding waste residue and collect to obtain filtrate, filtrate being concentrated, while recycling has
Solvent.It is 1 by volume and macroreticular resin quality by the concentrate that concentration is 2.0 mg/mL:1 ratio adds in D101 macropores
Static Adsorption is carried out in resin, adsorption time is 8 h, then upper prop.First with the distilled water of 2 times of column volumes and 2 times of column volumes
10% ethyl alcohol removes impurity.Then eluted again with 20% ethanol solution, collect 20% ethanol elution part.Eluent is concentrated,
200.6 g of Bergenin crude product is obtained, Bergenin sterling 169.7g is obtained after 95% ethyl alcohol recrystallization.
The yield of Bergenin crude product detects purity more than 80% 2% or so, by HPLC, by 95% ethyl alcohol
Bergenin sterling can be obtained after recrystallization, yield rock in 1.7% or so, the treebine stem harvested by different sources, season is white
The content difference of dish element is larger, and yield is higher, relatively low belongs to normal phenomenon.This law is using the repeatable macroreticular resin utilized and again
The method that crystallization combines prepares Bergenin, has the advantages that operation is succinct, at low cost, is easy to industrialization amplification.
4 macroreticular resin Study on selection of embodiment
Macroreticular resin has the polytypes such as polarity, low pole, nonpolar, and in the present invention, inventor has selected LSA-
700（Polarity）、LSA-700B（Polarity）、D941（Polarity）、AB-8（Low pole）、D3520（It is nonpolar）、D101（It is nonpolar）This
7 kinds of macroreticular resins are as object is investigated, by static white to determine to be suitable for rock in purifying treebine stem with Dynamic Adsorption, desorption
The macroreticular resin of dish element.
Staticadsorption experiment results are as shown in Figure 2, it is seen that LSA-700, LSA-700B, AB-8, D101, D941, D3520 six
Kind of macroreticular resin adsorbs after 24 h of Static Adsorption close to saturation, Static Adsorption rate more than 90 %, wherein AB-8,
The static saturated adsorption rate of tri- kinds of resins of D101, D3520 is higher than other 3 kinds.Require resin adsorption amount big in production and desorption
Rate is high, to ensure that active ingredient recycles to greatest extent.Therefore inventor is further using the desorption experiment of macroreticular resin come preferably
Resin, the results are shown in Figure 3 for experiment.
As can be seen from Figure 3, above-mentioned six kinds of macroreticular resins show apparent otherness when static state desorbs：Wherein D101, AB-
8th, the desorption ability of D3520 resins is stronger, more than 80 %, is suitable for isolating and purifying for Bergenin in treebine stem；And
The desorption ability of LSA-700, LSA-700B, D941 resin is weaker, only about 30 %, and unfavorable raw material returns in purification process
It receives, causes wastage of material, therefore do not select.
In addition, show that these three resins of D101, AB-8, D3520 inhale the dynamic saturation of Bergenin by dynamic experiment
Attached amount, dynamic desorption rate are suitable, and without notable difference, therefore inventor selects this 3 kinds of macroreticular resins of D101, AB-8, D3520 to use
Bergenin isolates and purifies in treebine stem.
The selection optimization of 5 concentration of alcohol of embodiment and dosage
Using thin-layer chromatography（TLC）Ingredient in different concentration ethanol efflux is detected, the condition of solvent is chlorine
It is imitative:Ethyl acetate:Methanol=5:4:2.5, the results are shown in Table 1.As shown in Table 1, the ethanol elution macropore of distilled water and 10 %
It during resin, has no Bergenin and is washed out, be mainly at this time water-solubility impurity in eluent；When the ethyl alcohol that eluent is 20 %
When, it is tracked according to TLC, discovery has Bergenin to be eluted.It is other while purple bergenia herb is eluted with the increase of concentration of alcohol
Oil-soluble impurities also flows out therewith, and the purifying for being unfavorable for Bergenin also results in waste of solvent, cost increases, therefore in subsequent experimental
In further investigate concentration of alcohol for 20 %, 40 %, 60 % when elution profile, result is as shown in table 1,2,3.
It can be seen that first with 2 times of column volumes（BV）The ethanol solution elution of water, 2 times of column volumes 10% can remove impurity elimination
Matter, then eluted with the ethanol water of 7 times of column volumes 20%, obtained Bergenin crude product purity is high, for AB-8, D101,
Tri- kinds of macroreticular resins of D3520 are respectively 84.98%, 88.13%, 82.18%.
The optimization of 6 sample concentration of embodiment
When extraction finishes, after carrying out separation of solid and liquid, the amount of Bergenin is certain in extracting solution, but the journey of extracting solution concentration
Degree directly influences the concentration of Bergenin in extracting solution, and the concentration of Bergenin is to have an impact to macroporous resin adsorption, desorption
, therefore inventor investigates the influence of sample concentration.Table 4,5,6 is when Determination of Bergenin is 39.94 mg in extracting solution
Experimental result.
From above-mentioned experimental result as it can be seen that when sample concentration is 1.997 mg/ml 2 mg/ml of ≈, the absorption of macroreticular resin
Amount and desorption efficiency are optimal, therefore when concentration for being concentrated into Bergenin is about 2 mg/mL carries out loading（It is carried out with liquid chromatography
Detection）.
Macroreticular resin can become larger to the adsorption rate of Bergenin in extracting solution with the growth of adsorption time, and inventor exists
It being found in Staticadsorption experiment as 8 more than h of adsorption time, adsorption rate amplitude of variation is little, and in view of the production cycle, therefore
Adsorption time is set to 8 h.
Claims (3)
- A kind of 1. method of the extraction purification Bergenin from treebine stem, it is characterised in that include the following steps：（1）Treebine stem is crushed, is sieved, is 1 according to mass volume ratio：8 ~ 20 add in 40 ~ 60% ethanol solution refluxing extraction；（2）Separation of solid and liquid obtains filtrate, concentrates the filtrate to the 1/2 ~ 1/3 of its original volume, while recycling design;（3）When into concentrate, addition macroreticular resin Static Adsorption 6 ~ 8 is small, macroreticular resin is selected from AB-8, D3520, D101；（4）By the macroreticular resin upper prop after absorption, impurity elimination is removed with the distilled water of 2 times of column volumes and 10% ethyl alcohol of 2 times of column volumes Matter；（5）With 20% ethanol elution of 7 times of column volumes, 20% ethanol eluate is collected；（6）20% ethanol eluate is concentrated, obtains Bergenin crude product；（7）Bergenin crude product is carried out being recrystallized to give Bergenin sterling with 95% ethyl alcohol.
- 2. method according to claim 1, it is characterised in that：Step（2）In, the method for separation of solid and liquid is filtering or centrifugation point From filtrate is concentrated into 2 mg/mL of Bergenin concentration.
- 3. method according to claim 1, it is characterised in that：Step（6）In, the method for concentration for be concentrated in vacuo or Film concentrates.
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| CN106632373A (en) * | 2016-09-30 | 2017-05-10 | 陕西科技大学 | A D crystal form solid substance of bergenin, a preparing method thereof and uses of the substance |
| CN106749298B (en) * | 2016-11-18 | 2019-03-29 | 文县嘉诚农副产品有限公司 | A kind of Bergenin extracts and refining methd |
| CN110272433A (en) * | 2018-08-12 | 2019-09-24 | 长沙爱扬医药科技有限公司 | A method of extracting quercitrin and Bergenin |
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