CN114470034A - Preparation method of liquorice crude extract - Google Patents
Preparation method of liquorice crude extract Download PDFInfo
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- CN114470034A CN114470034A CN202210128914.0A CN202210128914A CN114470034A CN 114470034 A CN114470034 A CN 114470034A CN 202210128914 A CN202210128914 A CN 202210128914A CN 114470034 A CN114470034 A CN 114470034A
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- licorice extract
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/484—Glycyrrhiza (licorice)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
Abstract
The invention discloses a preparation method of a liquorice crude extract, belonging to the field of traditional Chinese medicine extraction and comprising the following steps: pulverizing Glycyrrhrizae radix with low temperature liquid nitrogen, and sieving to obtain Glycyrrhrizae radix powder; adding water into the licorice powder, stirring uniformly, soaking, adding biological enzyme and nonionic active agent for enzymolysis; inactivating enzyme, leaching, filtering, and concentrating to obtain concentrated solution; eluting the concentrated solution, collecting the eluent, and drying to obtain the crude licorice extract. The preparation method is simple, has high extraction efficiency, high purity and high yield of the crude licorice extract, really achieves the extraction of active substances, efficiently separates medicinal active ingredients from impurity ingredients, and can furthest retain the activity of the crude licorice extract.
Description
Technical Field
The invention relates to the field of traditional Chinese medicine extraction, in particular to a preparation method of a liquorice crude extract.
Background
The liquorice is one of common Chinese herbal medicines, has the functions of adrenocortical hormone, resisting inflammation, ulcer and anaphylactic reaction, resisting cancer, bacteria and virus, promoting pancreatic secretion, inhibiting isolated intestine, regulating immunologic function, relieving cough, eliminating phlegm, resisting mutation, detoxifying, resisting oxidation, protecting ear and vestibule functions, promoting urination, protecting liver, preventing arteriosclerosis, resisting cerebral ischemia, preventing diabetic complications and the like.
The licorice extract is a component extracted from licorice and having medicinal value. The licorice extract generally comprises: glycyrrhizin, glycyrrhizic acid, liquiritin, glycyrrhetin, semaphorin, formononetin, quercetin, etc. As a yellow to brown-yellow powder. Has the effects of invigorating spleen and replenishing qi, clearing away heat and toxic materials, eliminating phlegm and relieving cough, relieving spasm and pain, and harmonizing the medicines. Can be used for treating weakness of spleen and stomach, asthenia, palpitation, short breath, cough, excessive phlegm, abdominal distention, and spasm and pain of limbs.
At present, the extraction method of the commonly used licorice extract mainly comprises a water boiling alcohol precipitation method, a microwave extraction method, an alkaline extraction method, an enzymatic extraction method, an ultrafiltration method, an ultrasonic extraction method and the like. However, the extraction methods in the prior art have the defects of low extraction rate, low purity, high cost, unstable properties and the like in the extraction process, and cannot adapt to the development trend of the modern active ingredient extraction technology.
Disclosure of Invention
The invention aims to provide a preparation method of a liquorice crude extract, which aims to solve the problems in the prior art.
In order to achieve the purpose, the invention provides the following scheme:
the invention provides a preparation method of a liquorice crude extract, which comprises the following steps:
(1) pulverizing Glycyrrhrizae radix with low temperature liquid nitrogen, and sieving to obtain Glycyrrhrizae radix powder;
(2) adding water into the licorice powder obtained in the step (1), uniformly stirring and soaking, and adding a biological enzyme and a nonionic active agent for enzymolysis;
(3) carrying out enzyme deactivation, leaching, filtering and concentrating on the solution subjected to enzymolysis in the step (2) to obtain a concentrated solution;
(4) and (4) eluting the concentrated solution obtained in the step (3), collecting the eluent, and drying to obtain the compound.
Further, in the low-temperature liquid nitrogen crushing process in the step (1), the feeding amount of nitrogen is 1-3 mL/s, and the feeding amount of liquorice is 2-4 g/s; the crushing temperature is-10-0 ℃;
preferably, the mixture is ground and then sieved by a sieve of 100-120 meshes.
Further, the mass ratio of the licorice powder, the water, the biological enzyme and the nonionic surfactant in the step (2) is 1: 3-5: 0.08-0.1: 0.1.
Further, the biological enzyme in the step (2) is one or more of cellulase, pectinase, protease and alpha-amylase;
preferably, the nonionic surfactant is one or more of polysorbate, polyoxyethylene fatty acid ester, and polyoxyethylene fatty alcohol ether.
Further, the enzymolysis in the step (2) is carried out for 0.5-1.5 h under the conditions that the temperature is 50-60 ℃ and the pH value is 4-6.
Further, the enzyme deactivation in the step (3) is carried out for 5-15 min at the temperature of 80-100 ℃.
Further, the leaching in the step (3) is extracting for 60-90 min at the temperature of 80-100 ℃;
preferably, the concentration in the step (3) is vacuum concentration, and specifically, the enzymolysis liquid is concentrated to 1/3-1/5 of the original volume at reduced pressure and low temperature.
Further, the elution in the step (4) is to adsorb the concentrated solution on macroporous resin, and sequentially use distilled water and 70-90% ethanol for elution.
Furthermore, the amount of the distilled water is 3-5 times of the volume of the column, and the distilled water is eluted until the column is colorless;
preferably, the dosage of the ethanol is 20-40 times of the column volume.
Further, the drying in the step (4) is drying for 2-5 hours under a vacuum condition at-50-30 ℃.
Compared with the prior art, the invention has the following technical effects:
the method adopts low-temperature liquid nitrogen crushing combined with a nonionic surfactant assisted enzyme method to extract the crude licorice extract, has the advantages of easily available raw materials, simple process and convenient operation, and is suitable for large-scale industrial production. The method of the invention reduces the extraction times, improves the yield and purity of the crude licorice extract, reduces the production cost, maintains the stable structure and property of the crude licorice extract, and greatly improves the trophism, safety and acceptability of the crude licorice extract. The nonionic surfactant has the promotion effect on cellulase and pectinase in the hydrolysis process. Shortens the pretreatment and extraction time, reduces the time cost and improves the economic benefit.
The method can furthest retain the effective components of the liquorice, has simple preparation method, high extraction efficiency and high purity and yield of the liquorice crude extract, really achieves the extraction of active substances, efficiently separates medicinal active components from impurity components, and furthest retains the activity of the liquorice crude extract.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
Example 1
A method for preparing radix astragali crude extract comprises the following steps:
(1) pulverizing dried and clean Glycyrrhrizae radix with low temperature liquid nitrogen; sieving with 150 mesh sieve to obtain Glycyrrhrizae radix powder; wherein the feeding amount of nitrogen is 2mL/s and the feeding amount of liquorice is 3g/s in the low-temperature liquid nitrogen crushing process; the crushing temperature is-5 ℃;
(2) performing enzymolysis treatment on 1 part of astragalus powder, 4 parts of water, 0.1 part of polysorbate, 0.04 part of cellulase, 0.02 part of pectinase, 0.02 part of protease and 0.02 part of alpha-amylase for 1 hour at the temperature of 55 ℃ and the pH value of 5.0;
(3) inactivating enzyme of the enzymolysis solution at 90 deg.C for 10 min; extracting at 90 deg.C for 75 min; filtering; concentrating 1/4 of the original volume of the enzymolysis liquid at low temperature under vacuum to obtain a concentrated solution;
(4) adsorbing the concentrated solution on macroporous resin adsorption column, eluting with 4 times of distilled water to colorless, eluting with 30 times of 80% ethanol, and drying at-10 deg.C under vacuum for 3 hr.
Example 2
A method for preparing radix astragali crude extract comprises the following steps:
(1) pulverizing dried and clean Glycyrrhrizae radix with low temperature liquid nitrogen; sieving with 100 mesh sieve to obtain Glycyrrhrizae radix powder; wherein the feeding amount of nitrogen is 1mL/s and the feeding amount of licorice is 2g/s in the low-temperature liquid nitrogen crushing process; the crushing temperature is-10 ℃;
(2) carrying out enzymolysis treatment on 1 part of astragalus powder, 4 parts of water, 0.1 part of polyoxyethylene fatty acid ester active agent, 0.04 part of cellulase, 0.02 part of pectinase, 0.01 part of protease and 0.01 part of alpha-amylase for 0.5h at the temperature of 50 ℃ and the pH value of 4.0;
(3) inactivating enzyme of the enzymolysis solution at 80 deg.C for 5 min; extracting at 80 deg.C for 60 min; filtering; concentrating 1/3 of the original volume of the enzymolysis liquid at low temperature under vacuum to obtain a concentrated solution;
(4) adsorbing the concentrated solution on macroporous resin adsorption column, eluting with 3 times of distilled water to colorless, eluting with 20 times of 70% ethanol, and drying at-50 deg.C under vacuum for 2 hr.
Example 3
A method for preparing radix astragali crude extract comprises the following steps:
(1) pulverizing dried and clean Glycyrrhrizae radix with low temperature liquid nitrogen; sieving with 200 mesh sieve to obtain Glycyrrhrizae radix powder; wherein the feeding amount of nitrogen is 3mL/s and the feeding amount of liquorice is 4g/s in the low-temperature liquid nitrogen crushing process; the crushing temperature is 0 ℃;
(2) performing enzymolysis treatment on 1 part of astragalus powder, 4 parts of water, 0.1 part of polyoxyethylene fatty alcohol ether activator, 0.04 part of cellulase, 0.02 part of pectinase, 0.01 part of protease and 0.02 part of alpha-amylase at the temperature of 60 ℃ and the pH value of 6.0 for 1.5 hours;
(3) inactivating enzyme of the enzymolysis solution at 100 deg.C for 15 min; then extracting at 100 deg.C for 90 min; filtering; concentrating 1/5 of the original volume of the enzymolysis liquid at low temperature under vacuum to obtain a concentrated solution;
(4) adsorbing the concentrated solution on macroporous resin adsorption column, eluting with 5 times of distilled water to colorless, eluting with 40 times of 90% ethanol, and drying at 30 deg.C under vacuum for 5 hr.
In the case of the example 4, the following examples are given,
the difference from the embodiment 1 is that the feeding amount of nitrogen in the low-temperature liquid nitrogen crushing process in the step (1) is 1mL/s, and the feeding amount of liquorice is 2 g/s; the crushing temperature was-10 ℃.
Example 5
The difference from example 1 is that step (2) does not contain a protease.
Example 6
The difference from example 1 is that step (2) does not contain alpha-amylase.
Example 6
The difference from the example 1 is that the enzymolysis in the step (2) is carried out for 1h under the conditions that the temperature is 60 ℃ and the pH value is 4.8.
Example 7
The difference from example 1 is that the enzyme deactivation in step (3) is carried out at a temperature of 100 ℃ for 9 min.
Example 8
The difference from example 1 is that the leaching in step (3) is carried out at a temperature of 80 ℃ for 80 min.
Example 9
The difference from example 1 is that the concentration in step (3) is 4/15 which is vacuum reduced and low temperature concentrated enzymolysis liquid concentrated into original volume.
Example 10
The difference from example 1 is that in step (4), 90% ethanol solution was used as the ethanol solution in the amount of 30 column volumes.
Example 11
The difference from example 1 is that the drying in step (4) was carried out under vacuum at-30 ℃ for 2 hours.
Comparative example 1
Extracting according to conventional extraction method.
Comparative test
The total flavone content of the crude licorice extracts of examples 1 to 3 and comparative example 1 was determined, and the results are shown in Table 1.
TABLE 1 comparison of total flavone content of crude extract of radix Glycyrrhizae in different preparation methods
As a result: as can be seen from Table 1, the licorice extract produced by the preparation method of the present invention has a total flavone content of more than 2.9%, which is higher than that of the extract produced by the comparative example, and further verifies the feasibility and reliability of the process improvement of the present invention.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Claims (10)
1. A preparation method of a crude licorice extract is characterized by comprising the following steps: the method comprises the following steps:
(1) pulverizing Glycyrrhrizae radix with low temperature liquid nitrogen, and sieving to obtain Glycyrrhrizae radix powder;
(2) adding water into the licorice powder obtained in the step (1), uniformly stirring and soaking, and adding a biological enzyme and a nonionic active agent for enzymolysis;
(3) carrying out enzyme deactivation, leaching, filtering and concentrating on the solution subjected to enzymolysis in the step (2) to obtain a concentrated solution;
(4) and (4) eluting the concentrated solution obtained in the step (3), collecting the eluent, and drying to obtain the compound.
2. The method for preparing crude licorice extract according to claim 1, wherein the crude licorice extract is prepared by the following steps: in the step (1), the feeding amount of nitrogen is 1-3 mL/s and the feeding amount of liquorice is 2-4 g/s in the low-temperature liquid nitrogen crushing process; the crushing temperature is-10-0 ℃;
preferably, the mixture is ground and then sieved by a sieve of 100-120 meshes.
3. The method for preparing crude licorice extract according to claim 1, wherein the crude licorice extract is prepared by the following steps: in the step (2), the mass ratio of the licorice powder to the water to the biological enzyme to the nonionic surfactant is 1: 3-5: 0.08-0.1: 0.1.
4. The method for preparing crude licorice extract according to claim 3, wherein the crude licorice extract is prepared by the following steps: the biological enzyme in the step (2) is one or more of cellulase, pectinase, protease and alpha-amylase;
preferably, the nonionic surfactant is one or more of polysorbate, polyoxyethylene fatty acid ester, and polyoxyethylene fatty alcohol ether.
5. The method for preparing crude licorice extract according to claim 1, wherein the crude licorice extract is prepared by the following steps: in the step (2), enzymolysis is carried out for 0.5-1.5 h under the conditions that the temperature is 50-60 ℃ and the pH value is 4-6.
6. The method for preparing crude licorice extract according to claim 1, wherein the crude licorice extract is prepared by the following steps: and (3) inactivating the enzyme at 80-100 ℃ for 5-15 min.
7. The method for preparing crude licorice extract according to claim 1, wherein the crude licorice extract is prepared by the following steps: the leaching in the step (3) is carried out for 60-90 min at the temperature of 80-100 ℃;
preferably, the concentration in the step (3) is vacuum concentration, and specifically, the enzymolysis liquid is concentrated to 1/3-1/5 of the original volume at reduced pressure and low temperature.
8. The method for preparing crude licorice extract according to claim 1, wherein the crude licorice extract is prepared by the following steps: and (4) in the step (4), the concentrated solution is adsorbed on macroporous resin, and is sequentially eluted by using distilled water and 70-90% of ethanol.
9. The method for preparing crude extract of glycyrrhiza uralensis according to claim 8, wherein the method comprises the following steps: the using amount of the distilled water is 3-5 times of the column volume, and the distilled water is eluted until the column is colorless;
preferably, the dosage of the ethanol is 20-40 times of the column volume.
10. The method for preparing crude licorice extract according to claim 1, wherein the crude licorice extract is prepared by the following steps: and the drying in the step (4) is drying for 2-5 hours under the vacuum condition of-50-30 ℃.
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| CN202210128914.0A CN114470034A (en) | 2022-02-11 | 2022-02-11 | Preparation method of liquorice crude extract |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115368338A (en) * | 2022-08-26 | 2022-11-22 | 中国热带农业科学院香料饮料研究所 | Method for extracting piperine from fresh pepper fruits |
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| CN103142682A (en) * | 2013-02-28 | 2013-06-12 | 惠州市九惠制药股份有限公司 | Method for extracting liquorice flavonoids from liquorice residue |
| CN103613674A (en) * | 2013-09-09 | 2014-03-05 | 华南农业大学 | Extraction method for astragalus polysacharin |
| CN108210555A (en) * | 2018-01-25 | 2018-06-29 | 安徽诚亚生物科技有限公司 | A kind of homogenate extraction method of glycyrrhiza total flavonoid |
| CN111214535A (en) * | 2018-11-26 | 2020-06-02 | 南京西博恩生物科技有限公司 | A method for extracting Glycyrrhrizae radix extract from Glycyrrhrizae radix |
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- 2022-02-11 CN CN202210128914.0A patent/CN114470034A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103142682A (en) * | 2013-02-28 | 2013-06-12 | 惠州市九惠制药股份有限公司 | Method for extracting liquorice flavonoids from liquorice residue |
| CN103613674A (en) * | 2013-09-09 | 2014-03-05 | 华南农业大学 | Extraction method for astragalus polysacharin |
| CN108210555A (en) * | 2018-01-25 | 2018-06-29 | 安徽诚亚生物科技有限公司 | A kind of homogenate extraction method of glycyrrhiza total flavonoid |
| CN111214535A (en) * | 2018-11-26 | 2020-06-02 | 南京西博恩生物科技有限公司 | A method for extracting Glycyrrhrizae radix extract from Glycyrrhrizae radix |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115368338A (en) * | 2022-08-26 | 2022-11-22 | 中国热带农业科学院香料饮料研究所 | Method for extracting piperine from fresh pepper fruits |
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