CN101962386A

CN101962386A - Process for extracting arteannuin by biological compound enzymes

Info

Publication number: CN101962386A
Application number: CN2010105085614A
Authority: CN
Inventors: 肖文中; 黄凯
Original assignee: 肖文中
Priority date: 2010-10-15
Filing date: 2010-10-15
Publication date: 2011-02-02

Abstract

The invention discloses a process for extracting arteannuin by biological compound enzymes, and in the process, high-purity arteannuin is extracted from sweet wormwood leaves by biological compound enzyme preparations, which greatly improves the extraction yield of the arteannuin. The process is characterized by comprising the following steps: spraying a certain amount of compound enzyme solution on dry sweet wormwood leaves, and poling up the dry sweet wormwood leaves so as to decompose cell walls, fibrous tissues and plant wax oils in the sweet wormwood leaves by enzymes; then extracting arteannuin by organic solvent, and carrying out concentration and crystallization on the obtained arteannuin; finally, carrying out re-crystallization and drying on the obtained arteannuin by appropriate solvent so as to obtain the high-purity arteannuin. Compared with the traditional methods, in the invention, the finished product ratio can be enhanced by 10 percent, and the process of purification by using column chromatography also can be avoided, thereby greatly simplifying the production process and significantly reducing the consumption of organic solvent.

Description

The compound bio-enzyme method is extracted the technology of Artemisinin

Technical field

The present invention relates to a kind of method of from sweet wormwood, extracting the high purity Artemisinin with the compound bio-enzyme method.

Background technology

Artemisinin is a kind of efficient, the safe anti-cruel medicine that extracts from the Herba Artemisiae annuae of feverfew artemisia (Artemisiaannua L is commonly called as sweet wormwood).

The production method of Artemisinin mainly contains industrial naptha method, acetone extraction-column chromatography, rare pure method, methanol extraction-petroleum ether extraction method, supercritical extraction, microwave extraction method etc. at present.More than these methods all have to a certain degree problem, if any extract yield low (traditional method is about 80%), the production cycle that has is long, solvent loss is big, what have has particular requirement to equipment, production cost is high.

Summary of the invention

Equipment used of the present invention is simple, and Production Flow Chart is short, and solvent consumption is few, and production cost is low, and yield significantly improves than traditional method.

The present invention adopts biological enzymolysis technology, destroys cell walls etc. and makes the Artemisinin extraction yield improve 10%, destroys the wax oil composition, has avoided the operation of dewaxing or column chromatography, suitable solvent extract concentrate get final product the Artemisinin crystallization.

The present invention is achieved by the following technical programs:

The compound bio-enzyme method is extracted the technology of Artemisinin, at first at 0.1%～0.3% compound bio-enzyme of sweet wormwood foliage spray sweet wormwood weight, is deposited in the darkroom of 45 ± 2 ℃ of temperature, covers a layer of plastic film above the heap head and makes it to ferment 1～2 day.Fermentation enzymolysis good sweet wormwood blade is dried or dry; The sweet wormwood cured leaf is dropped in the extractor, add the solvent orange 2 A of 5 times of amounts at every turn, extract 3 times, each 2 hours, extract 60 ± 5 ℃ of temperature; United extraction liquid filters, and filtrate is concentrated into 5%～10% of original volume, and reclaims solvent orange 2 A; Concentrated solution leaves standstill, and natural cooling crystallization treats that solution temperature is below 30 ℃ the time, the leaching crystallization is also collected filtrate, and filter cake (crude product crystallization) is drained after solvent orange 2 A washs 2 times with extracting, and goes in the baking oven dry, filtrate is dropped in the extractor, makes next series-produced extraction solvent.The crystallization of exsiccant crude product with 100 times of amount solvent B dissolving, is filtered, be concentrated into original volume 10% and reclaim solvent B; Concentrated solution leaves standstill crystallisation by cooling, treats that solution temperature is below 30 ℃ the time, and the leaching crystallization is also collected mother liquor, and crystallization is drained with solvent B washing 3 times, goes in the baking oven dryly, promptly gets the Artemisinin of content more than 99%; Mother liquor is according to the Artemisinin that gets with the concentrated repeatedly cooling crystallization of method more than 99%.

Described compound bio-enzyme is included as cellulase, lipase, two or more combination such as polygalacturonase.

Described solvent orange 2 A is sherwood oil (30～60,60～90,90～120), No. six solvent oils, No. 120 gasoline, normal hexane, ethyl acetate etc.

Described solvent orange 2 A solvent B is an alcohol in high concentration.

Beneficial effect:

The present invention has broken away from the general method extracting the overcritical of environment, and can effectively improve the extraction yield of Artemisinin, make yield rate improve about 10%, and avoid using column chromatography purification than traditional method, simplify production technique greatly, and can significantly reduce the consumption of organic solvent.

Embodiment

Give an actual example below and describe the present invention.

Embodiment one is the Artemisinin extraction to the low levels grass:

Sweet wormwood grass cured leaf 500kg (artemislnin content 0.49%), evenly spray the certain density solution of 800g compound bio-enzyme preparation, be deposited in the darkroom of 44 ℃ of temperature, cover layer of plastic film above the heap head and make it to ferment 2 days, blade takes out during for black and dries, in the input multi-function extractor, extract 3 times for 65 ℃ with ethyl acetate, each 2hr, extract solvent load 2500L/ time, united extraction liquid is discharged to crude product crystallizer natural cooling crystallization when being concentrated into about 300L, during 30 ℃ of monitoring crystallization solutions, put a jar suction filtration and get crude product crystallization 2.74kg.Produce 3 batches with quadrat method regeneration.

Throw sweet wormwood 2013kg altogether for 4 batches, get crude product crystallization 10.1kg, use the 1000L95% dissolve with ethanol, filter, filtrate is concentrated into 100L, put to elaboration crystallizer crystallisation by cooling, leaching needle crystal during 30 ℃ of crystallization solutions of monitoring, mother liquor is concentrated into half of original volume, places crystallization equally, so quadruplication is separated out most Artemisinin crystallizations.Merge Artemisinin crystallization oven dry, weigh 9.02kg, content 99.0%.Artemisinin yield 90.5%.

Embodiment two is the Artemisinin extraction to the moderate content grass:

Sweet wormwood grass cured leaf 503kg (artemislnin content 0.74%) evenly sprays the certain density solution of 600g compound bio-enzyme preparation, is deposited in the darkroom of 45 ℃ of temperature, covers layer of plastic film above the heap head and makes it to ferment 1 day half.Blade takes out during for black and dries, in the input multi-function extractor, extract 3 times for 63 ℃ with No. six solvent oils, each 2hr, extract solvent load 2500L/ time, united extraction liquid is discharged to crude product crystallizer natural cooling crystallization when being concentrated into about 500L, during 30 ℃ of monitoring crystallization solutions, put a jar suction filtration and get crude product crystallization 3.74kg.Produce 3 batches with quadrat method regeneration.

Throw sweet wormwood 2005kg altogether for 4 batches, get crude product crystallization 15.0kg, use the 1500L90% dissolve with ethanol, filter, filtrate is concentrated into 150L, put to elaboration crystallizer crystallisation by cooling, leaching needle crystal behind the 8hr, mother liquor is concentrated into half of original volume, places crystallization equally, so quadruplication is separated out most Artemisinin crystallizations.Merge Artemisinin crystallization oven dry, weigh 13.5kg, content 99.4%.Artemisinin yield 90.4%.

Embodiment three is the extraction to the Artemisinin of high-content grass:

Sweet wormwood grass cured leaf 501kg (artemislnin content 0.93%) evenly sprays the certain density solution of 500g compound bio-enzyme preparation, is deposited in the darkroom of 44 ℃ of temperature, covers layer of plastic film above the heap head and makes it to ferment 1 day.Blade takes out during for black and dries, in the input multi-function extractor, extract 3 times for 65 ℃ with No. 120 gasoline, each 2hr, extract solvent load 2500L/ time, united extraction liquid is discharged to crude product crystallizer natural cooling crystallization when being concentrated into about 500L, during 30 ℃ of monitoring crystallization solutions, put a jar suction filtration and get crude product crystallization 4.42kg.Produce 3 batches with quadrat method regeneration.

Throw sweet wormwood 2008kg altogether for 4 batches, get crude product crystallization 18.2kg, use the 1800L95% dissolve with ethanol, filter, filtrate is concentrated into 180L, put to elaboration crystallizer crystallisation by cooling, leaching needle crystal during 30 ℃ of crystallization solutions of monitoring, mother liquor is concentrated into half of original volume, places crystallization equally, so quadruplication is separated out most Artemisinin crystallizations.Merge Artemisinin crystallization oven dry, weigh 17.1kg, content 99.5%.Artemisinin yield 91.1%.

More than show and described ultimate principle of the present invention and principal character and advantage of the present invention.The technician of the industry should understand; the present invention is not restricted to the described embodiments; that describes in the foregoing description and the specification sheets just illustrates principle of the present invention; under the prerequisite that does not break away from smart kind of the present invention and scope; the present invention also has various changes and modifications, and these changes and improvements all fall in the claimed scope of the invention.The claimed scope of the present invention is defined by appending claims and equivalent thereof.

Claims

1. the compound bio-enzyme method is extracted the technology of Artemisinin, it is characterized in that, is undertaken by following step:

A) at 0.1%～0.3% compound bio-enzyme of sweet wormwood foliage spray sweet wormwood weight, be deposited in the darkroom of 45 ± 2 ℃ of temperature, cover a layer of plastic film above the heap head and make it to ferment 1～2 day;

B) the good sweet wormwood blade that will ferment enzymolysis dries or dries;

C) the sweet wormwood cured leaf is dropped in the extractor, add the solvent orange 2 A of 5 times of amounts at every turn, extract 3 times, each 2 hours, extract 60 ± 5 ℃ of temperature;

D) united extraction liquid filters, and filtrate is concentrated into 5%～10% of original volume, and reclaims solvent orange 2 A;

E) with d) concentrated solution leave standstill, natural cooling crystallization is when treating that solution temperature is 30 ± 1 ℃; The leaching crystallization is also collected filtrate, and filter cake (crude product crystallization) is used c) in extraction solvent orange 2 A washing 2 times after drain, send in the baking oven dryly, filtrate is dropped in the extractor, makes next series-produced extraction solvent;

F) with e) crystallization of exsiccant crude product dissolves with 100 times of solvent B, filtration, be concentrated into original volume 10% and reclaim solvent B;

G) with f) in concentrated solution leave standstill crystallisation by cooling, when treating that solution temperature is 30 ± 1 ℃, the leaching crystallization is also collected mother liquor, crystallization is drained with solvent B washing 3 times, sends in the baking oven dryly, promptly gets the Artemisinin of content more than 99%; Mother liquor is according to the Artemisinin that gets with the concentrated repeatedly cooling crystallization of method more than 99%.

2. compound bio-enzyme method according to claim 1 is extracted the technology of Artemisinin, it is characterized in that described compound bio-enzyme is included as cellulase, lipase, two or more combination formula such as polygalacturonase.

3. compound bio-enzyme method according to claim 1 is extracted the technology of Artemisinin, it is characterized in that described solvent orange 2 A is a sherwood oil, No. six solvent oils, No. 120 gasoline, normal hexane, ethyl acetate etc.; Solvent B is an alcohol in high concentration.