CN102911055A - Method for extracting chlorogenic acid - Google Patents
Method for extracting chlorogenic acid Download PDFInfo
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- CN102911055A CN102911055A CN2012104533167A CN201210453316A CN102911055A CN 102911055 A CN102911055 A CN 102911055A CN 2012104533167 A CN2012104533167 A CN 2012104533167A CN 201210453316 A CN201210453316 A CN 201210453316A CN 102911055 A CN102911055 A CN 102911055A
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- chlorogenic acid
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- acid extracting
- japanese honeysuckle
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Abstract
The invention discloses a method for extracting a chlorogenic acid. The method includes the following steps of smashing honeysuckle, soaking smashed honeysuckle by ethanol or water, then performing wall-breaking extraction through ultrasound, microwave or enzyme, performing ethanol extraction to obtain a coarse extract, then concentrating the obtained coarse extract to obtain an extractum, adding water to dissolve the extractum to obtain supernatant, then adding the supernatant in a resin adsorbing column, washing by water, washing the column by a 10% to 50% (v/v) ethanol solution, collecting an eluant, enabling the eluant to pass through an activated charcoal column, and collecting a liquid to obtain the chlorogenic acid. The method for extracting the chlorogenic acid has the advantages that the preparation process is simple, the cost is low, the extraction efficiency of the chlorogenic acid is high, the content of the chlorogenic acid in prepared product is high, and the industrialized production is facilitated.
Description
Technical field
The present invention relates to a kind of technique, particularly the method for chlorogenic acid extracting.
Background technology
Japanese Honeysuckle belongs to caprifoliaceae plant, has clearing heat and detoxicating effect, be China commonly used also be one of historical longer Chinese medicine.Multiplex in the heating for the treatment of diseases caused by dampness, the various diseases such as common cold due to wind-heat clinically.Chlorogenic acid is one of effective constituent in the Japanese Honeysuckle, has multiple pharmacologically active as removing free radical, antisepsis and anti-inflammation, antiviral etc., therefore chlorogenic acid more and more is subject to medicine scholars' attention, and modern science has been deep into a plurality of fields such as food, health care, medicine and daily-use chemical industry to the research of chlorogenic acid pharmacologically active.
Itself has unstable chlorogenic acid, will avoid high temperature, long-time heating and strong illumination during extraction.The traditional method of chlorogenic acid extracting has the decocting cooking method from Japanese Honeysuckle, water extraction and alcohol precipitation method, rare alcohol reflux method, supercritical CO
2Extraction process, percolation etc.Supercritical CO
2The extraction process solvent is easy to get, and productive rate is high, but cost is also high, does not also have at present large-scale application in suitability for industrialized production.The percolation extraction efficiency is higher, but consuming time longer, and economic benefit is not high.The method of tradition separation and purification chlorogenic acid mainly contains extraction process, alkaline precipitation, Thin-layer chromatography, macroreticular resin absorbing method etc., alkaline precipitation easily makes the chlorogenic acid hydrolysis, the extraction rule needs a large amount of organic solvents, and Thin-layer chromatography and macroreticular resin absorbing method cost are higher, are unfavorable for suitability for industrialized production.
Summary of the invention
One of purpose of the present invention is to provide the method for chlorogenic acid extracting, and technique is simple, and cost is low, and extraction efficiency is high, and the chlorogenic acid purity that makes is high, is convenient to suitability for industrialized production.
For achieving the above object, technical scheme is:
The method of chlorogenic acid extracting comprises the steps:
A. use alcohol immersion after Japanese Honeysuckle being pulverized, then broken wall lixiviate makes crude extract;
B. step a gained crude extract is concentrated into medicinal extract, after being dissolved in water, gets supernatant liquor;
C. step b gained supernatant liquor being added in the resin absorbing column, after the water flushing, is that 10% ~ 50% ethanolic soln is washed post with volume fraction again, collects elutriant, and elutriant is crossed activated carbon column, and collection liquid gets chlorogenic acid.
Preferably, described step a is that to add volume fraction after Japanese Honeysuckle is pulverized be 60% ~ 80% ethanol, and pH transfers to 3 ~ 5, soaks then broken wall lixiviate 24-72 hour.
Preferred, described step a is that to add volume fraction after Japanese Honeysuckle is pulverized be 70% ethanol, and pH transfers to 4, soaks then broken wall lixiviate 24 hours.
Preferred, described broken wall lixiviate is under 50 ℃ ~ 70 ℃ conditions, and ultrasonic extraction twice each 30 minutes, merges extracted twice liquid.
Preferred, described broken wall lixiviate is to process 1 ~ 3 minute under output rating is the microwave condition of 600 W, is to extract twice under the 70-90 ℃ of condition with ethanol in temperature, 90 minutes for the first time, 30 minutes for the second time, merges extracted twice liquid.
Preferably, described step a replaces with following steps: add entry after Japanese Honeysuckle is pulverized, soaked 24-72 hour, then add the polygalacturonase that is equivalent to Japanese Honeysuckle weight 0.2% ~ 0.3%, the cellulase that is equivalent to Japanese Honeysuckle weight 0.5% ~ 0.6%, enzymolysis is 2.5 hours under 35 ℃ ~ 45 ℃ conditions, then uses twice of extraction using alcohol under 80 ℃ of conditions, each 2 hours, merge extracted twice liquid.
Preferably, described step b is concentrated into medicinal extract with step a gained crude extract, adds to be equivalent to 5-15 times of water dissolution of medicinal extract volume.
Preferably, described step c adds supernatant liquor in the resin absorbing column, after the water flushing with 5-10 times of column volume, be that 10% ~ 50% ethanolic soln is washed post with the volume fraction of 5-10 times of column volume again, collect elutriant, the elutriant anhydrous alcohol solution, supernatant liquor is crossed activated carbon column, collect liquid.
Beneficial effect of the present invention: the invention provides the method for chlorogenic acid extracting from Japanese Honeysuckle, its technique is simple, and extraction cost is low, in the leaching process Japanese Honeysuckle is pulverized, and has increased the surface-area that extracts, and is conducive to abundant extraction, improves the rate of recovery.Adopt ultrasonic extraction, Microwave-Assisted Ethanol extraction method and enzymolysis Assisted Ethanol extraction method in the leaching process, cytoclasis is abundant, guaranteed extraction efficiency, in order to obtain highly purified chlorogenic acid, extract is crossed first resin column, make with extra care with charcoal absorption, the chlorogenic acid purity that makes is high, is up to 85.2% again.
Embodiment
Below in conjunction with specific embodiment such scheme is described further.Should be understood that these embodiment are not limited to limit the scope of the invention for explanation the present invention.The implementation condition that adopts among the embodiment can be done further adjustment according to the condition of concrete producer, and not marked implementation condition is generally the condition in the normal experiment.
Embodiment 1
The method of chlorogenic acid extracting comprises the steps:
A. after Japanese Honeysuckle being pulverized, add volume fraction and be 60% ethanol, the feed liquid weight ratio is 1:10, and pH transfers to 3, soaked 72 hours, and then under 50 ℃ of conditions, ultrasonic extraction twice, each 30 minutes, merging extracted twice liquid made crude extract;
B. step a gained crude extract is concentrated into medicinal extract, adds the water dissolution that is equivalent to 10 times of medicinal extract weight, get supernatant liquor; Repeat 3 times, merge supernatant liquor, concentrated;
C. step b gained concentrated solution is added in the resin absorbing column (the preferred HPD100 model of adsorption column macroporous resin column), after 5 times of cylinder ponding flushings, be that 10% ethanolic soln is washed post with the volume fraction of 5 times of column volumes again, collect elutriant, elutriant is crossed activated carbon column, collect elutriant, get chlorogenic acid.
In the present embodiment, the rate of recovery of chlorogenic acid is 18.2%, and purity is 61.0%.
Embodiment 2
The method of chlorogenic acid extracting comprises the steps:
A. after Japanese Honeysuckle being pulverized, add volume fraction and be 80% ethanol, the feed liquid weight ratio is 1:20, and pH transfers to 4, soaked 36 hours, and then under 70 ℃ of conditions, ultrasonic extraction twice, each 30 minutes, merging extracted twice liquid made crude extract;
B. step a gained crude extract is concentrated into medicinal extract, adds the water dissolution that is equivalent to 5 times of medicinal extract weight, get supernatant liquor; Repeat 3 times, merge supernatant liquor, concentrated;
C. concentrated solution on the step b gained is added in the resin absorbing column (the preferred HPD100 model of adsorption column macroporous resin column), after 5 times of cylinder ponding flushings, be that 10% ethanolic soln is washed post with the volume fraction of 5 times of column volumes again, collect elutriant, elutriant is crossed activated carbon column, collect elutriant, get chlorogenic acid.
In the present embodiment, the rate of recovery of chlorogenic acid is 21.4%, and purity is 62.5%.
Embodiment 3
The method of chlorogenic acid extracting comprises the steps:
A. after Japanese Honeysuckle being pulverized, the adding volume fraction is 60% ethanol, the feed liquid weight ratio is 1:15, and pH transfers to 3, soaks 72 hours, be to process 3 minutes under the microwave condition of 600 W in output rating, be to extract twice under 70 ℃ of conditions with ethanol in temperature, 90 minutes for the first time, 30 minutes for the second time, merge extracted twice liquid, make crude extract;
B. step a gained crude extract is concentrated into medicinal extract, adds the water dissolution that is equivalent to 15 times of medicinal extract weight, get supernatant liquor; Repeat 3 times, merge supernatant liquor, concentrated;
C. concentrated solution on the step b gained is added in the resin absorbing column (the preferred HPD100 model of adsorption column macroporous resin column), after 8 times of cylinder ponding flushings, be that 20% ethanolic soln is washed post with the volume fraction of 8 times of column volumes again, collect elutriant, elutriant is crossed activated carbon column, collect elutriant, get chlorogenic acid.
In the present embodiment, the rate of recovery of chlorogenic acid is 29.1%, and purity is 70.6%.
Embodiment 4
The method of chlorogenic acid extracting comprises the steps:
A. after Japanese Honeysuckle being pulverized, add entry, the feed liquid weight ratio is 1:20, and pH transfers to 4, soaks 36 hours, be to process 1 minute under the microwave condition of 600 W in output rating, be to extract twice under 90 ℃ of conditions with ethanol in temperature, 90 minutes for the first time, 30 minutes for the second time, merge extracted twice liquid, make crude extract;
B. step a gained crude extract is concentrated into medicinal extract, adds the water dissolution that is equivalent to 5 times of medicinal extract weight, get supernatant liquor; Repeat 3 times, merge supernatant liquor, concentrated;
C. concentrated solution on the step b gained is added in the resin absorbing column (the preferred HPD100 model of adsorption column macroporous resin column), after 8 times of cylinder ponding flushings, be that 50% ethanolic soln is washed post with the volume fraction of 8 times of column volumes again, collect elutriant, elutriant is crossed activated carbon column, collect elutriant, get chlorogenic acid.
In the present embodiment, the rate of recovery of chlorogenic acid is 31.9%, and purity is 72.6%.
Embodiment 5
The method of chlorogenic acid extracting comprises the steps:
A. after Japanese Honeysuckle being pulverized, add entry, the feed liquid weight ratio is 1:10, soaked 72 hours, and then added the polygalacturonase that is equivalent to Japanese Honeysuckle weight 0.2% and the cellulase that is equivalent to Japanese Honeysuckle weight 0.6%, enzymolysis is 2.5 hours under 40 ℃ of conditions, then under 80 ℃ of conditions, use twice of extraction using alcohol, each 2 hours, merge extracted twice liquid, make crude extract;
B. step a gained crude extract is concentrated into medicinal extract, adds the water dissolution that is equivalent to 10 times of medicinal extract weight, get supernatant liquor; Repeat 3 times, merge supernatant liquor, concentrated;
C. concentrated solution on the step b gained is added in the resin absorbing column (the preferred HPD100 model of adsorption column macroporous resin column), after 5 times of cylinder ponding flushings, be that 20% ethanolic soln is washed post with the volume fraction of 5 times of column volumes again, collect elutriant, elutriant is crossed activated carbon column, collect liquid, get chlorogenic acid.
In the present embodiment, the rate of recovery of chlorogenic acid is 34.6%, and purity is 78.0%.
Embodiment 6
The method of chlorogenic acid extracting comprises the steps:
A. after Japanese Honeysuckle being pulverized, the adding volume fraction is 80% ethanol, the feed liquid weight ratio is 1:15, soaked 36 hours, and then added the polygalacturonase that is equivalent to Japanese Honeysuckle weight 0.3% and the cellulase that is equivalent to Japanese Honeysuckle weight 0.5%, enzymolysis is 2.5 hours under 45 ℃ of conditions, then under 80 ℃ of conditions, use twice of extraction using alcohol, each 2 hours, merge extracted twice liquid, make crude extract;
B. step a gained crude extract is concentrated into medicinal extract, adds the water dissolution that is equivalent to 15 times of medicinal extract weight, get supernatant liquor; Repeat 3 times, merge supernatant liquor, concentrated;
C. concentrated solution on the step b gained is added in the resin absorbing column (the preferred HPD100 model of adsorption column macroporous resin column), after 5 times of cylinder ponding flushings, be that 20% ethanolic soln is washed post with the volume fraction of 5 times of column volumes again, collect elutriant, elutriant is crossed activated carbon column, collect liquid, get chlorogenic acid.
Present embodiment is most preferred embodiment of the present invention, and the rate of recovery of chlorogenic acid is 38.4%, and purity is 85.2%.
Explanation is at last, above embodiment is only unrestricted in order to technical scheme of the present invention to be described, although by invention has been described with reference to the preferred embodiments of the present invention, but those of ordinary skill in the art is to be understood that, can make various changes to it in the form and details, and not depart from the present invention that appended claims limits.
Claims (8)
1. the method for chlorogenic acid extracting is characterized in that, comprises the steps:
A. use alcohol immersion after Japanese Honeysuckle being pulverized, then broken wall lixiviate makes crude extract;
B. step a gained crude extract is concentrated into medicinal extract, after being dissolved in water, gets supernatant liquor;
C. step b gained supernatant liquor being added in the resin absorbing column, after the water flushing, is that 10% ~ 50% ethanolic soln is washed post with volume fraction again, collects elutriant, and elutriant is crossed activated carbon column, and collection liquid gets chlorogenic acid.
2. the method for described chlorogenic acid extracting according to claim 1 is characterized in that: described step a is that to add volume fraction after Japanese Honeysuckle is pulverized be 60% ~ 80% ethanol, and the feed liquid weight ratio is 1:10-20, and pH transfers to 3 ~ 5, soaks then broken wall lixiviate 24-72 hour.
3. the method for described chlorogenic acid extracting according to claim 2 is characterized in that: described step a is that to add volume fraction after Japanese Honeysuckle is pulverized be 70% ethanol, and the feed liquid weight ratio is 1:15, and pH transfers to 4, soaks then broken wall lixiviate 24 hours.
4. the method for each described chlorogenic acid extracting according to claim 1-3, it is characterized in that: described broken wall lixiviate is under 50 ℃ ~ 70 ℃ conditions, ultrasonic extraction twice each 30 minutes, merges extracted twice liquid.
5. the method for each described chlorogenic acid extracting according to claim 1-3, it is characterized in that: described broken wall lixiviate is to process 1 ~ 3 minute under output rating is the microwave condition of 600 W, be to extract twice under the 70-90 ℃ of condition with ethanol in temperature, 90 minutes for the first time, 30 minutes for the second time, merge extracted twice liquid.
6. the method for described chlorogenic acid extracting according to claim 1, it is characterized in that: described step a replaces with following steps: add entry after Japanese Honeysuckle is pulverized, soaked 24-72 hour, then add the polygalacturonase that is equivalent to Japanese Honeysuckle weight 0.2% ~ 0.3%, the cellulase that is equivalent to Japanese Honeysuckle weight 0.5% ~ 0.6%, enzymolysis is 2.5 hours under 35 ℃ ~ 45 ℃ conditions, then uses twice of extraction using alcohol under 80 ℃ of conditions, each 2 hours, merge extracted twice liquid.
7. the method for described chlorogenic acid extracting according to claim 1, it is characterized in that: described step b is concentrated into medicinal extract with step a gained crude extract, adds to be equivalent to 5-15 times of water dissolution of medicinal extract volume.
8. the method for described chlorogenic acid extracting according to claim 1, it is characterized in that: described step c adds supernatant liquor in the resin absorbing column, after the water flushing with 5-10 times of column volume, be that 20% ethanolic soln is washed post with the volume fraction of 5-10 times of column volume again, collect elutriant, the elutriant anhydrous alcohol solution is crossed activated carbon column with supernatant liquor, collects liquid.
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| CN201210453316.7A CN102911055B (en) | 2012-11-13 | 2012-11-13 | Method for extracting chlorogenic acid |
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| CN201210453316.7A CN102911055B (en) | 2012-11-13 | 2012-11-13 | Method for extracting chlorogenic acid |
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103086889A (en) * | 2013-02-22 | 2013-05-08 | 王星敏 | Method for extracting chlorogenic acid from Lonicera japonica leaves activated under catalysis of enzyme |
| CN103787885A (en) * | 2014-03-06 | 2014-05-14 | 山东禾本堂生物科技有限公司 | Method of extracting chlorogenic acid with high purity from honeysuckle above ground part |
| CN104788314A (en) * | 2015-04-03 | 2015-07-22 | 广西大学 | Method for extracting chlorogenic acid from honeysuckle flowers |
| CN105085257A (en) * | 2015-06-24 | 2015-11-25 | 四川农业大学 | Method for extracting heavy chlorogenic acid from Eupatorium adenophorum Spreng |
| CN106071622A (en) * | 2016-06-15 | 2016-11-09 | 谢晓亮 | Clear honeysuckle solid beverage and preparation method thereof |
| CN108586241A (en) * | 2018-01-17 | 2018-09-28 | 浙江树人学院 | The method of ultrasonic-microwave-aqueous two-phase collaboration extraction separation sweet potato leaves chlorogenic acid |
| CN108653370A (en) * | 2018-07-31 | 2018-10-16 | 庐山市环绿时代农业科技开发有限公司 | A kind of preparation method of honeysuckle particle |
| CN109809997A (en) * | 2019-03-07 | 2019-05-28 | 四川君正生物饲料有限公司 | A method of the chlorogenic acid extracting from coffee grounds |
| CN110305007A (en) * | 2019-07-25 | 2019-10-08 | 广西民族大学 | One kind extracting caffeinic method from a silk fabric |
| CN111153759A (en) * | 2020-01-17 | 2020-05-15 | 金陵药业股份有限公司 | Method for extracting total phenolic acid from waste liquid of ester extraction in Mailuoning injection production |
| CN113875844A (en) * | 2021-09-29 | 2022-01-04 | 武夷山成隆天创茶业科技股份有限公司 | Application of spartina alterniflora extract in plant-derived functional tea product and functional tea product |
| CN114031498A (en) * | 2021-11-05 | 2022-02-11 | 临沂市农业科学院 | Method for extracting high-purity honeysuckle chlorogenic acid by membrane separation method |
| CN114751825A (en) * | 2022-05-24 | 2022-07-15 | 河北瑞龙生物科技有限公司 | Method for efficiently extracting chlorogenic acid from honeysuckle |
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| CN101314568A (en) * | 2008-06-24 | 2008-12-03 | 南京工业大学 | Novel method for adsorption separation of high purity chlorogenic acid |
| CN101838200A (en) * | 2009-03-20 | 2010-09-22 | 上海中医药大学 | Method for extracting and separating chlorogenic acid from honeysuckle |
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Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103086889B (en) * | 2013-02-22 | 2015-11-25 | 王星敏 | A kind of method of enzyme catalysis activation leaf of Flos Lonicerae chlorogenic acid extracting |
| CN103086889A (en) * | 2013-02-22 | 2013-05-08 | 王星敏 | Method for extracting chlorogenic acid from Lonicera japonica leaves activated under catalysis of enzyme |
| CN103787885A (en) * | 2014-03-06 | 2014-05-14 | 山东禾本堂生物科技有限公司 | Method of extracting chlorogenic acid with high purity from honeysuckle above ground part |
| CN104788314A (en) * | 2015-04-03 | 2015-07-22 | 广西大学 | Method for extracting chlorogenic acid from honeysuckle flowers |
| CN105085257A (en) * | 2015-06-24 | 2015-11-25 | 四川农业大学 | Method for extracting heavy chlorogenic acid from Eupatorium adenophorum Spreng |
| CN106071622B (en) * | 2016-06-15 | 2020-03-20 | 河北省农林科学院经济作物研究所 | Clear honeysuckle solid beverage and preparation method thereof |
| CN106071622A (en) * | 2016-06-15 | 2016-11-09 | 谢晓亮 | Clear honeysuckle solid beverage and preparation method thereof |
| CN108586241B (en) * | 2018-01-17 | 2021-03-05 | 浙江树人学院 | Method for extracting and separating chlorogenic acid of sweet potato leaves by ultrasonic-microwave-aqueous two-phase cooperation |
| CN108586241A (en) * | 2018-01-17 | 2018-09-28 | 浙江树人学院 | The method of ultrasonic-microwave-aqueous two-phase collaboration extraction separation sweet potato leaves chlorogenic acid |
| CN108653370A (en) * | 2018-07-31 | 2018-10-16 | 庐山市环绿时代农业科技开发有限公司 | A kind of preparation method of honeysuckle particle |
| CN109809997A (en) * | 2019-03-07 | 2019-05-28 | 四川君正生物饲料有限公司 | A method of the chlorogenic acid extracting from coffee grounds |
| CN110305007A (en) * | 2019-07-25 | 2019-10-08 | 广西民族大学 | One kind extracting caffeinic method from a silk fabric |
| CN111153759A (en) * | 2020-01-17 | 2020-05-15 | 金陵药业股份有限公司 | Method for extracting total phenolic acid from waste liquid of ester extraction in Mailuoning injection production |
| CN113875844A (en) * | 2021-09-29 | 2022-01-04 | 武夷山成隆天创茶业科技股份有限公司 | Application of spartina alterniflora extract in plant-derived functional tea product and functional tea product |
| CN114031498A (en) * | 2021-11-05 | 2022-02-11 | 临沂市农业科学院 | Method for extracting high-purity honeysuckle chlorogenic acid by membrane separation method |
| CN114751825A (en) * | 2022-05-24 | 2022-07-15 | 河北瑞龙生物科技有限公司 | Method for efficiently extracting chlorogenic acid from honeysuckle |
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