CN107674084A - A kind of preparation method of high-purity qinghaosu - Google Patents
A kind of preparation method of high-purity qinghaosu Download PDFInfo
- Publication number
- CN107674084A CN107674084A CN201711142254.7A CN201711142254A CN107674084A CN 107674084 A CN107674084 A CN 107674084A CN 201711142254 A CN201711142254 A CN 201711142254A CN 107674084 A CN107674084 A CN 107674084A
- Authority
- CN
- China
- Prior art keywords
- qinghaosu
- sweet wormwood
- preparation
- purity
- silica gel
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
- C07D493/12—Heterocyclic compounds containing oxygen atoms as the only ring hetero atoms in the condensed system in which the condensed system contains three hetero rings
- C07D493/20—Spiro-condensed systems
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention relates to sweet wormwood production technical field, and in particular to a kind of preparation method of high-purity qinghaosu.The inventive method comprises the following steps：S1. sweet wormwood petroleum ether extract is prepared, adjusts solid-liquid ratio and temperature to 25 40 DEG C；S2. take silica gel to fill post, control silica gel amount of fill, loading volume and loading flow velocity in every silica gel, complete the absorption of sample loading；S3. the mixed liquor of petroleum ether and ethyl acetate is taken to be eluted as eluent；S4. post eluent is concentrated at 40 85 DEG C, and the volume after concentration is 1,/10 1/20 before concentration；S5. qinghaosu crystallizes.A kind of preparation method of high-purity qinghaosu is provided, the inventive method is suitable for the removal of sweet wormwood alkene and impurity B in sweet wormwood extract in big production, and technique is controllable, and the content by sweet wormwood alkene in qinghaosu of efficiency is less than 2%, and the content of impurity B is less than 0.2%.
Description
【Technical field】
The present invention relates to sweet wormwood production technical field, and in particular to a kind of preparation method of high-purity qinghaosu.
【Background technology】
Qinghaosu is that to be extracted from compound inflorescence plant Artemisia annua cauline leaf have the sesquiterpene lactone medicine of peroxy-radical, by
Chinese pharmacy man slaughter cry of a deer found that there is the characteristics of less toxic, efficient, quick-acting in 1971.Qinghaosu is after pyrimethamine, chlorine
Maximally effective antimalarial specific drug after quinoline, primary quinoline, especially for encephalic malaria and anti-chlorine quinoline malaria, have quick-acting and less toxic
Feature, it is " the only effective malaria treatment medicine in the world " to be once referred to as by the World Health Organization.The anti-malarial mechanism of action is main
The process being in treatment malaria is activated by qinghaosu produces free radical, free radical and malaria original protein binding, acts on malaria original
The film structure of worm, its vacuolar membrane, nuclear membrane and plasma membrane is set to be destroyed, mitochondrial swelling, interior outer membrane comes off, so as to former to malaria
The eucaryotic cell structure and its function of worm damage.
According to the statistics of World Health Organization, from 2000, the population of Sub-Saharan Africa area about 2.4 hundred million was benefited from
Qinghaosu conjoint therapy, about 1,500,000 people are dead caused by the therapy avoids malaria.Therefore, many African common people address respectfully it
For " east god's medicine ".In addition artemisinine also shows that tempting prospect in the treatment of other diseases.Such as anti-schistosome, regulation or
Suppress the immunologic function of body fluid, improve the conversion ratio of lymphocyte, cholagogic, eliminating the phlegm, antibechic, relieving asthma.
Current anti-malarial qinghaosu related preparations in the world, the overwhelming majority is injection.Because injection is directly injected into
In human body or in blood, if the impurity content exceeding index in medicine, allergy or other lesions are easily caused to even result in death, institute
To continue to increase with China the limitation to relative substance in injection in the world at present, in intermediate the requirement of impurity also constantly carry
Height, impurity requirement is also improved constantly in other drugs.Sweet wormwood alkene is listed in impurity in qinghaosu extract solution intermediate, because
This in qinghaosu extraction process control sweet wormwood alkene amount, be qinghaosu extractive technique the most important thing.
Sweet wormwood alkene, Artemisitene, molecular formula C15H20O5, with the molecular formula and chemical structural formula of qinghaosu all very
It is close, large-scale production process is particularly during extraction purification and is difficult to remove, turns into the problem of sweet wormwood extraction, at present on how to go
Except the research of sweet wormwood alkene in Artemisinin is also in blank stage.Sweet wormwood alkene is shown as with high performance liquid chromatography detection process
In, a miscellaneous peak before qinghaosu, based on sweet wormwood alkene, it can be brought for the qinghaosu for being mainly used as injection serious
Adverse reaction.Sweet wormwood alkene and qinghaosu it can not such as separate well, then the purity of qinghaosu will be reduced, bring serious peace
Full hidden danger.The content of sweet wormwood alkene in existing commercially available at present or outlet qinghaosu product, using area normalization method, sweet wormwood alkene contains
Measure between 2.5-3.5%.
Chinese antimalarial artemisinin, it is the dry product leaf by annual herb plant sweet wormwood, uses petroleum ether, acetic acid second
Ester, alcohol etc. are as solvent extraction gained.The source of impurity B is mainly controlled not by being brought into extraction process with operating process
When other compositions are unstable to be transformed, and impurity B is shown as during with high performance liquid chromatography detection, in qinghaosu
A preceding miscellaneous peak, composition need to be further confirmed that, although little from peak area, because composition is unknown, for master
To be used as that for the qinghaosu of injection serious adverse reaction can be brought.Such as impurity B and qinghaosu can not be separated well,
The purity of qinghaosu will be so reduced, brings severe compromise.It is miscellaneous in existing commercially available at present or outlet qinghaosu product
Matter B content, using area normalization method, the content of impurity B is 0.5-0.9%.
【The content of the invention】
The goal of the invention of the present invention is：A kind of the problem of in background technology, there is provided preparation of high-purity qinghaosu
Method, the inventive method are adapted to the removal of sweet wormwood alkene and impurity B in sweet wormwood extract big production, and technique is controllable,
The content by sweet wormwood alkene in qinghaosu of efficiency is less than 2%, and the content of impurity B is less than 0.2%.
To achieve these goals, the technical solution adopted by the present invention is as follows：
A kind of preparation method of high-purity qinghaosu, comprises the following steps：
S1. sweet wormwood petroleum ether extract is taken, the solid-liquid ratio of the extract solution and sweet wormwood crude drug is 1：6-20kg/L, it is quiet
Put, it is standby when the temperature of liquid to be extracted is down to 25-40 DEG C；
S2. silica gel dress post is taken to obtain silica gel splitter, the amount of fill of silica gel is 200-800kg in every silica gel splitter, so
The extract solution in step S1 is added on silica gel splitter afterwards and carried out, the sample-loading amount of the every silicagel column is 10000-
26000L extract solution, adjustable column subflow speed is 800-3500L/h, completes sample loading；
S3. take the mixed liquor of petroleum ether and ethyl acetate to be eluted as eluent, collect median elution liquid, obtain sweet wormwood
Eluent, it is standby；
S4. post eluent is taken to be concentrated, the temperature of the concentration is 45-75 DEG C, and the volume after concentration is before concentrating
1/8-1/20, stand still for crystals, take out crystal drying, obtain crude product, it is standby；
S5. crude product is taken, then toward alcohol is added in crude product, dissolves, is filtered successively with diatomite and miillpore filter respectively, stand
Crystallization, takes crystalline solid to produce qinghaosu.
Optimization, calculated using area normalization method, the content of sweet wormwood alkene is less than 2% in qinghaosu, and the content of impurity B is low
In 0.2%.
More optimize, the preparation method of the sweet wormwood petroleum ether extract is following any：
A. take Artemisia annua to be added in extractor, add petroleum ether heating and refluxing extraction 1-3 times, each 30-150 points
Clock, filtrate is filtered to take, merging filtrate obtains petroleum ether extract, adjusts solid-liquid ratio, produces；
B. Artemisia annua is taken by solvent extraction, after recycling design, with petroleum ether dissolution, is adjusted solid-liquid ratio, is produced.
Further optimize, the solid-liquid ratio of the extract solution and sweet wormwood crude drug is 10-15kg/L；The every silicagel column
Sample-loading amount be 12000-25000L extract solution；The flow velocity is 1500-2500L/h.
Further optimize, the volume ratio of the eluent petrochina ether and ethyl acetate is 93-99：1-7.
Further optimize again, the dosage of the eluent is 5-10 times of column volume.
Further optimize again, the aperture of the miillpore filter is 0.22-0.45 microns.
Further optimize again, the temperature control of crystallization is at 13-22 DEG C.
Further optimize again, method for crystallising is specially in S5：Alcohol is added into crude product, is dissolved, with diatomite and micro-
Hole filter membrane is filtered successively, and alcohol is added into liquid to be crystallized, is stood, is obtained crystal I and filtrate I；Filtrate I is taken to continue to concentrate,
Inward plus alcohol, then filtered successively with diatomite and miillpore filter, stand, obtain crystal II and filtrate II；Filtrate is by above-mentioned
If the number that method is concentrated crystallization is 1-3 times；Merge the crystal obtained every time.
Further optimize, the temperature control of crystallization is at 25-48 DEG C.
Further optimize again, adding ethanol toward crude product is crystallized, and the mass fraction of the ethanol is 80-95%,
The dosage of the ethanol is 5-15 times of crude product.
In summary, by adopting the above-described technical solution, the beneficial effects of the invention are as follows：
1. present invention firstly provides being studied for how to remove sweet wormwood alkene in qinghaosu, by molten to extract solution
Agent, temperature, pillar filling species, dosage etc. are screened, and have obtained being suitable for through agent, efficient purification process；By tradition
The content of sweet wormwood alkene is substantially 3% or so in the qinghaosu that method purification process obtains, and the sweet wormwood of the inventive method after purification
Alkene content is less than 2%, can most preferably reach 0.93% in embodiment, more up to lower in actual production process, even below
0.5%.Sweet wormwood alkene is because chemical constitution and property and qinghaosu are very close, in lab scale process into large-scale production process, due to
The change of extraction purification amount is obtained with adjusting to optimum state, the present invention by constantly studying, it is necessary to tested many times
Purification process, be very suitable for removing sweet wormwood olefinic impurity content in qinghaosu in big production, save solvent, improve income.
2. ground in the present invention for the sample concentration of sample, loading volume, loading speed etc. in qinghaosu purifying
Study carefully, research in production is found, in silicagel column silica gel dosage number directly determine the size for separating sample, in production if
When silica gel dosage is less than 200kg in single post, the sample size of separation is few, material and time cost steeply rise increase by 30% with
On；And after loading is more than 800kg, column volume becomes big, even if expanding column diameter or increase length, during will also result in loading
Absorption is uneven, and pressure is excessive, has more than 15% to be in unadsorbed state in silica gel, separating effect is bad, and sweet wormwood alkene is miscellaneous from that need to make
Matter increases above more than 2.5%.
3. the present invention is dissolved the sample of loading with petroleum ether according to produce reality, regulation petroleum ether extract with
The solid-liquid ratio of sweet wormwood crude drug is 1：6-20kg/L, most preferably 10-15kg/L；What is so coordinated often want, and the applied sample amount of silica gel is
10000-26000L extract solution, pillar flow velocity are 800-3500L/h, while arrival efficiently separates, have taken into account solvent use again
Amount, production time, device requirement, it can be adapted to produce needs greatly.Because petroleum ether has very strong volatility, thus it is right before loading
Its temperature also has strict limitation, controls at 25-40 DEG C, reduces volatilization, reduces volumetric errors.
4. qinghaosu is determination system of thermal unstable material, because the formulation that qinghaosu is prepared into anti-malarial medicine is largely to inject
Based on agent, and at present it is internal and international on requirements at the higher level are all proposed to traditional Chinese medicine, the check criteria of impurity is constantly carried
Height, this requires the purity continuous improvement as the bulk drug of injection.Various chemicals can be used such as in the lab
The chemical reagent such as methanol, acetone purify to material can obtain purer material, and in production, it is desirable to except certain low impurity
Content, relation factor and pass through most economical, safe, feasible method as caused by finding impurity B in industrial production and carry out
Control is not easy very much.It is because close to reagent type, production safety, production time cost, yield and economic benefit etc. in production
Cut is closed.The present invention have found key factor caused by impurity B, rear impurity completed in conventional purification by constantly studying, exploring
B is still higher, condition during by controlling concentration, such as temperature, volume, crystallization operation, effectively can drop the content of impurity B
As little as less than 0.2%, 3 times or so are reduced, reduce further the content of impurity B in qinghaosu.
5. impurity B is one kind studies the less element of the first species at present, less to this kind of composition Study at present, mass spectrum point is carried out
After the core mass analysis such as analysis, it is determined that containing one of 9- isoarteannuins, other compositions need to be further determined that.The present invention is right first
Generation, the purifying of this constituents are studied, and the generation of this kind of material are reduced by strict technological operation；The work
Skill is not introduced into other reagents and compared with multi-step, economical, environmentally friendly, feasible, and has no to enter such material in items are reported at present
Row research.Because company is that current qinghaosu yield accounts for the world 1/4, caused good economic benefit and society after application of result
Benefit.
Reach the purpose of effectively control impurity B content 6. inventive method is comprehensive each process procedure, in a very long time,
Screening key factor is incomplete, is all difficult to reach the purpose of effectively control impurity B；And it is too many to screen key factor, some right and wrong
Key factor, influenceing less, conversely to cause the efficiency of production after control reduces, cost increase.
【Brief description of the drawings】
Fig. 1 embodiments 5 and the efficient liquid phase comparison diagram of control sample 1；
In accompanying drawing, the efficient liquid phase figure of A- embodiments 5, the efficient liquid phase figure of B- control samples 2,1- solution peak, 2- sweet wormwood alkene, 3-
Impurity B, 4- qinghaosus.
【Embodiment】
With reference to embodiment, the invention will be further described.
Embodiment 1
A kind of preparation method of high-purity qinghaosu, comprises the following steps：
S1. sweet wormwood petroleum ether extract is taken, the solid-liquid ratio of the extract solution and sweet wormwood crude drug is 1：6, stand, it is to be extracted
It is standby when the temperature of liquid is down to 25 DEG C；
Wherein, the preparation method of sweet wormwood petroleum ether extract is：Artemisia annua is taken by solvent extraction, after recycling design, is used
Petroleum ether dissolution, solid-liquid ratio is adjusted, is produced.
S2. silica gel dress post is taken to obtain silica gel splitter, the amount of fill of silica gel be 200kg in every silica gel splitter, then general
Extract solution in step S1 is added on silica gel splitter and carried out, and the sample-loading amount of the every silicagel column is 10000 extract solution,
Adjustable column subflow speed is 800L/h, completes sample loading；
S3. take the mixed liquor of petroleum ether and ethyl acetate to be eluted as eluent, collect median elution liquid, obtain sweet wormwood
Eluent, it is standby；The volume ratio of eluent petrochina ether and ethyl acetate is 93：7, the dosage of eluent is 5 times of column volumes.
S4. post eluent is taken to be concentrated, the temperature of the concentration is 75 DEG C, and the volume after concentration is 1/8 before concentration,
Obtain concentrate and obtain crude product, it is standby；
S5. crude product is taken, then toward alcohol is added in crude product, dissolves, is filtered successively with diatomite and miillpore filter respectively, micropore
The aperture of filter membrane is 0.45 micron.Stand still for crystals, the temperature control of crystallization takes crystalline solid to produce qinghaosu at 13 DEG C.Using face
Product normalization method calculates, and the content of sweet wormwood alkene is less than 2% in qinghaosu, and the content of impurity B is less than 0.2%.
Methanol is added toward crude product to be crystallized, dosage is 6 times of crude product.
Wherein, method for crystallising is specially：Alcohol is added into crude product, dissolves, was carried out successively with diatomite and miillpore filter
Filter, alcohol is added into liquid to be crystallized, is stood, is obtained crystal I and filtrate I；Take filtrate I to continue to concentrate, add alcohol inward, dissolve, so
Filtered successively with diatomite and miillpore filter afterwards, stand, obtain crystal II and filtrate II；Merge the crystal obtained every time, press
The above method is recrystallized.The temperature control of crystallization is at 25 DEG C.
Embodiment 2
A kind of preparation method of high-purity qinghaosu, comprises the following steps：
S1. sweet wormwood petroleum ether extract is taken, the solid-liquid ratio of the extract solution and sweet wormwood crude drug is 1：20kg/L, stand,
It is standby when the temperature of liquid to be extracted is down to 40 DEG C；
Wherein, the preparation method of sweet wormwood petroleum ether extract is：Take Artemisia annua to be added in extractor, add petroleum ether
Heating and refluxing extraction 3 times, 30 minutes every time, filtrate is filtered to take, merging filtrate obtains petroleum ether extract, adjusts solid-liquid ratio, i.e.,
；
S2. silica gel dress post is taken to obtain silica gel splitter, the amount of fill of silica gel be 800kg in every silica gel splitter, then general
Extract solution in step S1 is added on silica gel splitter and carried out, and the sample-loading amount of the every silicagel column is 25000L extraction
Liquid, adjustable column subflow speed is 2500L/h, completes sample loading；
S3. take the mixed liquor of petroleum ether and ethyl acetate to be eluted as eluent, collect median elution liquid, obtain sweet wormwood
Eluent, it is standby；The volume ratio of eluent petrochina ether and ethyl acetate is 99：1, the dosage of eluent is 10 times of column volumes.
S4. post eluent is taken to be concentrated, the temperature of the concentration is 45 DEG C, and the volume after concentration is 1/ before concentration
20, stand still for crystals, take out crystal drying, obtain crude product, it is standby；
S5. crude product is taken, then toward alcohol is added in crude product, dissolves, is filtered successively with diatomite and miillpore filter respectively, micropore
The aperture of filter membrane is 0.22-0.45 microns.Stand still for crystals, the temperature control of crystallization takes crystalline solid to produce qinghaosu at 22 DEG C.Adopt
Calculated with area normalization method, the content of sweet wormwood alkene is less than 2% in qinghaosu, and the content of impurity B is less than 0.2%.
Ethanol is added toward crude product to be crystallized, the mass fraction of the ethanol is 95%, and the dosage of the ethanol is crude product
5 times.
Wherein, method for crystallising is specially：Alcohol is added into crude product, dissolves, was carried out successively with diatomite and miillpore filter
Filter, alcohol is added into liquid to be crystallized, is stood, is obtained crystal I and filtrate I；Take filtrate I to continue to concentrate, add alcohol inward, dissolve, so
Filtered successively with diatomite and miillpore filter afterwards, stand, obtain crystal II and filtrate II；Filtrate II is taken to continue to concentrate, then
Filtered successively with diatomite and miillpore filter, add alcohol inward, stood, obtain crystal III and filtrate II I；Merging obtains every time
Crystal, recrystallized as stated above.The temperature control of crystallization is at 48 DEG C.
Embodiment 3
A kind of preparation method of high-purity qinghaosu, comprises the following steps：
S1. sweet wormwood petroleum ether extract is taken, the solid-liquid ratio of the extract solution and sweet wormwood crude drug is 1：10kg/L, stand,
It is standby when the temperature of liquid to be extracted is down to 28 DEG C；
Wherein, the preparation method of sweet wormwood petroleum ether extract is：Take Artemisia annua to be added in extractor, add petroleum ether
Heating and refluxing extraction 1 time, 150 minutes every time, filtrate is filtered to take, merging filtrate obtains petroleum ether extract, adjusts solid-liquid ratio, i.e.,
；
S2. silica gel dress post is taken to obtain silica gel splitter, the amount of fill of silica gel be 300kg in every silica gel splitter, then general
Extract solution in step S1 is added on silica gel splitter and carried out, and the sample-loading amount of the every silicagel column is 12000L extraction
Liquid, adjustable column subflow speed is 1500L/h, completes sample loading；
S3. take the mixed liquor of petroleum ether and ethyl acetate to be eluted as eluent, collect median elution liquid, obtain sweet wormwood
Eluent, it is standby；The volume ratio of eluent petrochina ether and ethyl acetate is 94：6, the dosage of eluent is 8 times of column volumes.
S4. post eluent is taken to be concentrated, the temperature of the concentration is 52 DEG C, and the volume after concentration is 1/ before concentration
12, stand still for crystals, take out crystal drying, obtain crude product, it is standby；
S5. crude product is taken, then toward methanol is added in crude product, is filtered successively with diatomite and miillpore filter respectively, micropore filter
The aperture of film is 0.22 micron.Stand still for crystals, the temperature control of crystallization takes crystalline solid to produce qinghaosu at 15 DEG C.Using area
Normalization method calculates, and the content of sweet wormwood alkene is less than 2% in qinghaosu, and the content of impurity B is less than 0.2%.
Ethanol is added toward crude product to be crystallized, the mass fraction of the ethanol is 85%, and the dosage of the ethanol is crude product
13 times.
Wherein, method for crystallising is specially：Alcohol is added into crude product, dissolves, was carried out successively with diatomite and miillpore filter
Filter, alcohol is added into liquid to be crystallized, is stood, is obtained crystal I and filtrate I；Take filtrate I to continue to concentrate, add alcohol inward, dissolve, so
Filtered successively with diatomite and miillpore filter afterwards, stand, obtain crystal II and filtrate II；Merge the crystal obtained every time, press
The above method is recrystallized.The temperature control of crystallization is at 15 DEG C.
Embodiment 4
A kind of preparation method of high-purity qinghaosu, comprises the following steps：
S1. sweet wormwood petroleum ether extract is taken, the solid-liquid ratio of the extract solution and sweet wormwood crude drug is 15kg/L, stands, treats
It is standby when the temperature of extract solution is down to 36 DEG C；
Wherein, the preparation method of sweet wormwood petroleum ether extract is：Take Artemisia annua to be added in extractor, add petroleum ether
Heating and refluxing extraction 2 times, 60 minutes every time, filtrate is filtered to take, merging filtrate obtains petroleum ether extract, adjusts solid-liquid ratio, i.e.,
.
S2. silica gel dress post is taken to obtain silica gel splitter, the amount of fill of silica gel be 750kg in every silica gel splitter, then general
Extract solution in step S1 is added on silica gel splitter and carried out, and the sample-loading amount of the every silicagel column is 25000L extraction
Liquid, adjustable column subflow speed is 2500L/h, completes sample loading；
S3. take the mixed liquor of petroleum ether and ethyl acetate to be eluted as eluent, collect median elution liquid, obtain sweet wormwood
Eluent, it is standby；The volume ratio of eluent petrochina ether and ethyl acetate is 95：5, the dosage of eluent is 7 times of column volumes.
S4. post eluent is taken to be concentrated, the temperature of the concentration is 65 DEG C, and the volume after concentration is 1/ before concentration
16, stand still for crystals, take out crystal drying, obtain crude product, it is standby；
S5. crude product is taken, then toward alcohol is added in crude product, dissolves, is filtered successively with diatomite and miillpore filter respectively, micropore
The aperture of filter membrane is 0.45 micron.Stand still for crystals, the temperature control of crystallization takes crystalline solid to produce qinghaosu at 19 DEG C.Using face
Product normalization method calculates, and the content of sweet wormwood alkene is less than 2% in qinghaosu, and the content of impurity B is less than 0.2%.
Ethanol is added toward crude product to be crystallized, the mass fraction of the ethanol is 93%, and the dosage of the ethanol is crude product
12 times.
Wherein, method for crystallising is specially：Alcohol is added into crude product, dissolves, was carried out successively with diatomite and miillpore filter
Filter, alcohol is added into liquid to be crystallized, is stood, is obtained crystal I and filtrate I；Take filtrate I to continue to concentrate, add alcohol inward, then use silicon
Diatomaceous earth and miillpore filter are filtered successively, are stood, are obtained crystal II and filtrate II；Merge the crystal obtained every time, by above-mentioned side
Method is recrystallized.The temperature control of crystallization is at 18 DEG C.
Embodiment 5
A kind of preparation method of high-purity qinghaosu, comprises the following steps：
S1. sweet wormwood petroleum ether extract is taken, the solid-liquid ratio of the extract solution and sweet wormwood crude drug is 1：13kg/L, stand,
It is standby when the temperature of liquid to be extracted is down to 29 DEG C；
Wherein, the preparation method of sweet wormwood petroleum ether extract is following any：Artemisia annua is taken to be added in extractor,
Add petroleum ether heating and refluxing extraction 2 times, 90 minutes every time, filter to take filtrate, merging filtrate obtains petroleum ether extract, regulation
Solid-liquid ratio, produce.
S2. silica gel dress post is taken to obtain silica gel splitter, the amount of fill of silica gel be 500kg in every silica gel splitter, then general
Extract solution in step S1 is added on silica gel splitter and carried out, and the sample-loading amount of the every silicagel column is 18000L extraction
Liquid, adjustable column subflow speed is 2000L/h, completes sample loading；
S3. take the mixed liquor of petroleum ether and ethyl acetate to be eluted as eluent, collect median elution liquid, obtain sweet wormwood
Eluent, it is standby；The volume ratio of eluent petrochina ether and ethyl acetate is 97：3, the dosage of eluent is 8 times of column volumes.
S4. post eluent is taken to be concentrated, the temperature of the concentration is 55 DEG C, and the volume after concentration is 1/ before concentration
16, stand still for crystals, take out crystal drying, obtain crude product, it is standby；
S5. crude product is taken, then toward alcohol is added in crude product, dissolves, is filtered successively with diatomite and miillpore filter respectively, micropore
The aperture of filter membrane is 0.22 micron.Stand still for crystals, the temperature control of crystallization takes crystalline solid to produce qinghaosu at 17 DEG C.Using face
Product normalization method calculates, and the content of sweet wormwood alkene is less than 2% in qinghaosu, and the content of impurity B is less than 0.2%.
Ethanol is added toward crude product to be crystallized, the mass fraction of the ethanol is 90%, and the dosage of the ethanol is crude product
9 times.
Wherein, method for crystallising is specially：Alcohol is added into crude product, dissolves, was carried out successively with diatomite and miillpore filter
Filter, alcohol is added into liquid to be crystallized, is stood, is obtained crystal I and filtrate I；Take filtrate I to continue to concentrate, add alcohol inward, then use silicon
Diatomaceous earth and miillpore filter are filtered successively, are stood, are obtained crystal II and filtrate II；Merge the crystal obtained every time, by above-mentioned side
Method is recrystallized.The temperature control of crystallization is at 32 DEG C.
Embodiment 6 contrasts implementation result
Using high-efficient liquid phase technique each sample is detected, calculating sweet wormwood alkene in qinghaosu according to area normalization method contains
Amount.
1. sample preparation：Sample 50mg or so is taken, it is accurately weighed, put in 10ml measuring bottles, add flowing phased soln and be diluted to
Scale, shake up, 0.45 μm of membrane filtration.
2. test sample：The qinghaosu sample for taking the method by embodiment 1-5 to be prepared is test specimen 1-5.In pressing
State's Authorization Notice No. is the artemisinin-like that the method for embodiment 1 in CN102219790B Green extraction process for artemisinin methods obtains
Product are control sample 1；Presently commercially available qinghaosu sample is taken to be detected to obtain control sample 2；Carry out as described in Example 5
Prepare, make the ratio of eluent into petroleum ether：Ethyl acetate=88：12, column volume is constant, and other method is same with embodiment 5,
Obtain control sample 3；Prepared as described in Example 5, the temperature that post eluent concentrates is changed to 90 DEG C, recycling design is to dense
1/3 before contracting, the temperature of crystallization operation is changed to 25 DEG C, other method is same with embodiment 5, obtains control sample 4；
3. sample detection condition：
Chromatographic column：The strong silica gel that closes of octadecylsilane is filler；
Column temperature：30℃；
Wavelength：216nm；
Mobile phase：Acetonitrile：Water=50：50；
Flow velocity：1.0ml/min；
Sampling volume：20μl；
Run time：25min
4. the confirmation of composition in collection of illustrative plates：Using qinghaosu and sweet wormwood alkene standard sample, dissolved with methanol, in same inspection
Under the conditions of survey, confirmed according to appearance time.
5. computational methods：Impurity sweet wormwood alkene (being called impurity A in the present invention) and impurity B are calculated using area normalization method
The content of sweet wormwood alkene and impurity B percentage composition.
The content of sweet wormwood alkene and impurity B counts in each sample of table 1
| Sweet wormwood alkene content % | Impurity B content % | |
| Test specimen 1 | 1.876 | 0.185% |
| Test specimen 2 | 1.418 | 0.164% |
| Test specimen 3 | 1.394 | 0.124% |
| Test specimen 4 | 1.083 | 0.132% |
| Test specimen 5 | 0.89% | 0.115% |
| Control sample 1 | 2.952% | 0.985% |
| Control sample 2 | 2.864% | 1.114% |
| Control sample 3 | 2.468% | 0.796% |
| Control sample 4 | 1.988% | 0.869% |
From table, it can be seen that the content of sweet wormwood alkene is respectively less than 2% in 1-5 of the embodiment of the present invention, and the content of impurity B is equal
Less than 0.2%.And the content for the refined rear sweet wormwood alkene of control sample that post is extracted, crossed with conventional method is still up to 3%, such as
Sweet wormwood alkene content in this experiment in control sample is above 2.8%, and 2.5% can be also higher than in the case of.
In addition, the present invention has also carried out a series of single factor test contrast experiment, such as 1. independent investigation post concentrates are finally dense
The volume of contracting is different, and the content of impurity B is also different, and this obtains multiple authentication in production, the final volume of concentration too as large as
Volume is accumulated more than 1/10 condensate precursor after final concentration, then impurity B content increase in crystallization process, and is less than after 1/20, then
Crystallization effect is undesirable, and yield is low, while other impurities content is high.2. in actual production process, crystallization temperature change also can
Generation to impurity B has an impact, different from temperature during crystallization, and speed caused by filtering and crystallization is relevant, the highly crystalline formation of temperature
Not exclusively, and crystallization temperature it is too low produce speed it is fast, impurity easily be wrapped, it is difficult to remove.
Described above is the detailed description for the present invention preferably possible embodiments, but embodiment is not limited to this hair
Bright patent claim, the equal change completed or modification change under the technical spirit suggested by all present invention, all should belong to
Cover the scope of the claims in the present invention.
Claims (10)
1. a kind of preparation method of high-purity qinghaosu, it is characterised in that comprise the following steps：
S1. sweet wormwood petroleum ether extract is taken, the solid-liquid ratio of the extract solution and sweet wormwood crude drug is 1：6-20kg/L, stand, treat
It is standby when the temperature of extract solution is down to 25-40 DEG C；
S2. silica gel dress post is taken to obtain silica gel splitter, the amount of fill of silica gel be 200-800kg in every silica gel splitter, then general
Extract solution in step S1 is added on silica gel splitter and carried out, and the sample-loading amount of the every silicagel column is 10000-26000L's
Extract solution, adjustable column subflow speed is 800-3500L/h, completes sample loading；
S3. take the mixed liquor of petroleum ether and ethyl acetate to be eluted as eluent, collect median elution liquid, obtain sweet wormwood elution
Liquid, it is standby；
S4. post eluent is taken to be concentrated, the temperature of the concentration is 45-75 DEG C, and the volume after concentration is the 1/8- before concentration
1/20, stand still for crystals, take out crystal drying, obtain crude product, it is standby；
S5. crude product is taken, then toward alcohol is added in crude product, dissolves, is filtered successively with diatomite and miillpore filter respectively, stand knot
Crystalline substance, crystalline solid is taken to produce qinghaosu.
2. the preparation method of a kind of high-purity qinghaosu according to claim 1, it is characterised in that using area normalization
Method calculates, and the content of sweet wormwood alkene is less than 2% in qinghaosu, and the content of impurity B is less than 0.2%.
A kind of 3. preparation method of high-purity qinghaosu according to claim 1, it is characterised in that the sweet wormwood petroleum ether
The preparation method of extract solution is following any：
A. take Artemisia annua to be added in extractor, add petroleum ether heating and refluxing extraction 1-3 times, each 30-150 minutes, mistake
Leaching filtrate, merging filtrate obtain petroleum ether extract, adjust solid-liquid ratio, produce；
B. Artemisia annua is taken by solvent extraction, after recycling design, with petroleum ether dissolution, is adjusted solid-liquid ratio, is produced.
4. the preparation method of a kind of high-purity qinghaosu according to claim 1, it is characterised in that the extract solution and green grass or young crops
The solid-liquid ratio of wormwood artemisia crude drug is 10-15kg/L；The sample-loading amount of the every silicagel column is 12000-25000L extract solution；It is described
Flow velocity is 1500-2500L/h.
A kind of 5. preparation method of high-purity qinghaosu according to claim 1, it is characterised in that stone in the eluent
The volume ratio of oily ether and ethyl acetate is 93-99：1-7.
A kind of 6. preparation method of high-purity qinghaosu according to claim 5, it is characterised in that the use of the eluent
Measure as 5-10 times of column volume.
A kind of 7. preparation method of high-purity qinghaosu according to claim 1 or 5, it is characterised in that the micropore filter
The aperture of film is 0.22-0.45 microns.
A kind of 8. preparation method of high-purity qinghaosu according to claim 1, it is characterised in that the temperature control of crystallization
At 13-22 DEG C.
9. the preparation method of a kind of high-purity qinghaosu according to claim 1, it is characterised in that method for crystallising has in S5
Body is：Alcohol is added into crude product, is dissolved, is filtered successively with diatomite and miillpore filter, alcohol is added into liquid to be crystallized, it is quiet
Put, obtain crystal I and filtrate I；Take filtrate I to continue to concentrate, add alcohol inward, dissolve, then with diatomite and miillpore filter successively
Filtered, stand, obtain crystal II and filtrate II；If the number that filtrate is concentrated crystallization as stated above is 1-3 times；Close
And the crystal obtained every time.
10. the preparation method of a kind of high-purity qinghaosu according to claim 1, it is characterised in that add second toward crude product
Alcohol is crystallized, and the mass fraction of the ethanol is 80-95%, and the dosage of the ethanol is 5-15 times of crude product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711142254.7A CN107674084A (en) | 2017-11-17 | 2017-11-17 | A kind of preparation method of high-purity qinghaosu |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711142254.7A CN107674084A (en) | 2017-11-17 | 2017-11-17 | A kind of preparation method of high-purity qinghaosu |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107674084A true CN107674084A (en) | 2018-02-09 |
Family
ID=61149879
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711142254.7A Pending CN107674084A (en) | 2017-11-17 | 2017-11-17 | A kind of preparation method of high-purity qinghaosu |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107674084A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109651391A (en) * | 2018-12-27 | 2019-04-19 | 广西仙草堂制药有限责任公司 | A method of qinghaosu is recycled from qinghaosu recrystallization waste material |
| CN111141846A (en) * | 2019-12-31 | 2020-05-12 | 广西仙草堂制药有限责任公司 | Method for measuring content of artemisinin in sweet wormwood herb |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103483350A (en) * | 2013-06-25 | 2014-01-01 | 重庆华方武陵山制药有限公司 | Ultrasonic reflux extraction method of artemisinin |
| CN104628739A (en) * | 2013-11-09 | 2015-05-20 | 青岛中人智业生物科技有限公司 | Extraction technology of artemisinin |
| CN107235988A (en) * | 2017-06-21 | 2017-10-10 | 禹州市天源生物科技有限公司 | A kind of extracting method of qinghaosu and Artemisitene |
-
2017
- 2017-11-17 CN CN201711142254.7A patent/CN107674084A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103483350A (en) * | 2013-06-25 | 2014-01-01 | 重庆华方武陵山制药有限公司 | Ultrasonic reflux extraction method of artemisinin |
| CN104628739A (en) * | 2013-11-09 | 2015-05-20 | 青岛中人智业生物科技有限公司 | Extraction technology of artemisinin |
| CN107235988A (en) * | 2017-06-21 | 2017-10-10 | 禹州市天源生物科技有限公司 | A kind of extracting method of qinghaosu and Artemisitene |
Non-Patent Citations (4)
| Title |
|---|
| 周凌云: "青蒿素纯化工艺的研究", 《中国民族民间医药》 * |
| 张颖: "青蒿素的提取工艺优化研究", 《北方园艺》 * |
| 欧阳广娜: "天然抗疟药青蒿素应用和分离纯化工艺进展", 《科技传播》 * |
| 谢莹: "青蒿中青蒿素的提取分离及蒿甲醚的制备", 《中国药业》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109651391A (en) * | 2018-12-27 | 2019-04-19 | 广西仙草堂制药有限责任公司 | A method of qinghaosu is recycled from qinghaosu recrystallization waste material |
| CN109651391B (en) * | 2018-12-27 | 2021-08-13 | 广西仙草堂制药有限责任公司 | Method for recovering artemisinin from artemisinin recrystallization waste |
| CN111141846A (en) * | 2019-12-31 | 2020-05-12 | 广西仙草堂制药有限责任公司 | Method for measuring content of artemisinin in sweet wormwood herb |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP3896050B1 (en) | Method for preparing cannabidiol by means of high-speed countercurrent chromatography separation and purification | |
| CN107746407A (en) | A kind of method of sweet wormwood alkene in reduction qinghaosu | |
| CN102133298B (en) | Two-aqueous-phase method for extracting total saponin from bittersweet herb | |
| CN112010738B (en) | Industrial method for producing cannabinoid compound by utilizing chromatographic separation | |
| CN107674084A (en) | A kind of preparation method of high-purity qinghaosu | |
| CN101255182B (en) | Extraction separating method for bupleurum root saponin b2 | |
| CN106317148A (en) | Method for extracting cordycepin from cordyceps militaris | |
| CN103130817B (en) | Bilobalide B compound and preparation method thereof | |
| CN104892708B (en) | From the method for Armillaria luteo-virens fructification prepare with scale adenosine chemical reference substance | |
| CN103145729B (en) | Bilobalide B compound and preparation method thereof | |
| CN107827904A (en) | A kind of purification process of qinghaosu | |
| CN102924545B (en) | A kind of enrichment of akebiasaponin D and purification process | |
| CN104262341A (en) | Method for extracting vasodilatation active compound rhynchophylline | |
| CN101955501B (en) | Method for extracting and separating high-purity salidroside in pilot scale | |
| CN105801641B (en) | A kind of novel lignanoid's glycosides compound and its preparation method and application | |
| CN109400566A (en) | A method of extracting separating high-purity amentoflavone from Rock lily plant | |
| CN111437284B (en) | Sodium aescinate pharmaceutical composition and preparation method thereof | |
| CN103130818B (en) | Bilobalide B compound and preparation method thereof | |
| CN107235988A (en) | A kind of extracting method of qinghaosu and Artemisitene | |
| CN109490448B (en) | Preparation method of digoxin standard substance | |
| CN104496783B (en) | A kind of isolation and purification method of 4 '-O-methyl Bavachalcone B monomer | |
| CN104292081B (en) | A kind of method extracting batatasins from plant Rhizoma Dioscoreae | |
| CN114853840B (en) | Preparation method and application of akebia stem saponin D bulk drug | |
| CN114014900B (en) | Method for extracting and separating linarin from chrysanthemum indicum stems | |
| CN105001190B (en) | The method and system of extraction separation Vicenin-2 from Desmodium styracifolium |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180209 |