CN112603932B - Method for rapidly separating triterpene and sterol in ganoderma lucidum alcohol extract - Google Patents

Method for rapidly separating triterpene and sterol in ganoderma lucidum alcohol extract Download PDF

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CN112603932B
CN112603932B CN202011525201.5A CN202011525201A CN112603932B CN 112603932 B CN112603932 B CN 112603932B CN 202011525201 A CN202011525201 A CN 202011525201A CN 112603932 B CN112603932 B CN 112603932B
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sterol
column
triterpene
ganoderma
alcohol extract
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CN112603932A (en
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蔡鸿飞
张琴
许文东
袁诚
刘菊妍
杨阳
刘春芳
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Guangzhou Hanfang Pharmaceutical Co ltd
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06Fungi, e.g. yeasts
    • A61K36/07Basidiomycota, e.g. Cryptococcus
    • A61K36/074Ganoderma
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00Drugs for disorders of the urinary system
    • A61P13/08Drugs for disorders of the urinary system of the prostate
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00Drugs for disorders of the metabolism
    • A61P3/06Antihyperlipidemics
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00Drugs for disorders of the metabolism
    • A61P3/08Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00Antineoplastic agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00Drugs for immunological or allergic disorders
    • A61P37/02Immunomodulators
    • A61P37/04Immunostimulants
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation 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
    • A61K2236/50Methods involving additional extraction steps
    • A61K2236/55Liquid-liquid separation; Phase separation

Abstract

The invention relates to the field of natural medicine extraction and separation, and discloses a method for quickly separating triterpene and sterol from a ganoderma lucidum alcohol extract. The method adopts polyamide resin column packing, and uses a solvent with a specific proportion for elution to obtain triterpenoid and sterol eluent in the ganoderma alcohol extract, and the eluent is concentrated and dried to obtain ganoderma triterpenoid and ganoderma sterol. The ganoderan and ganoderan have similar structures and polarities, are easy to dissolve in organic solvents, are difficult to separate, and have complicated traditional preparation process and high production cost. Aiming at the defects of the prior art, the invention provides a method for quickly separating ganoderma triterpene and sterol components, which has the advantages of simple and convenient operation, high separation accuracy, good reproducibility, high safety and lower cost and is suitable for industrial production.

Description

Method for rapidly separating triterpene and sterol in ganoderma lucidum alcohol extract
Technical Field
The invention relates to the field of natural medicine extraction and separation, in particular to a method for quickly separating triterpene and sterol in a ganoderma lucidum alcohol extract.
Background
Ganoderma lucidum (Ganoderma lucidum) is a rare medicinal fungus, belongs to the general name of Ganoderma lucidum and Ganoderma sinense of Ganoderma of Polyporaceae of Basidiomycetes, and has effects of invigorating spleen and replenishing qi, nourishing yin and strengthening body resistance, strengthening body resistance and consolidating constitution, and prolonging life. The ganoderma lucidum is rich in components, including ganoderma lucidum polysaccharide, triterpene and sterol components, nucleoside, protein, trace elements and the like, wherein the ganoderma lucidum triterpene and sterol components and the ganoderma lucidum polysaccharide are the main active substances. The modern medicine research shows that the ganoderma lucidum triterpenoids have the effects of resisting tumors, protecting the liver, reducing blood sugar, inhibiting angiotensin, resisting oxidation and the like; the ganoderic acid has effects of enhancing disease resistance of human body, reducing cholesterol concentration in blood, and preventing prostate diseases.
At present, the ganoderma lucidum polysaccharide and the ganoderma lucidum triterpene are researched more at home and abroad, the ganoderma lucidum polysaccharide is easy to dissolve in water, acid, alkali, salt and other solutions but not in alcohol, ether, acetone and other organic solvents, and the ganoderma lucidum triterpene is difficult to dissolve in water but easy to dissolve in the organic solvents, so the ganoderma lucidum polysaccharide and the ganoderma lucidum triterpene are easy to separate, and the separation process is simpler and more mature. However, the ganodermasol and the ganodermasol have similar structures and polarities, are easy to dissolve in an organic solvent, and have larger separation difficulty. The main components of the ganoderma lucidum alcohol extract are ganoderma lucidum triterpene and ganoderma lucidum sterol, the research on ganoderma lucidum sterol substances is not deep enough at present, and the research report on the separation of the ganoderma lucidum sterol substances is very few, so that firstly, the content of the ganoderma lucidum sterol is relatively low, and secondly, the traditional preparation process is complicated and the production cost is high, so that a simple and convenient method for quickly separating the ganoderma lucidum triterpene and the ganoderma lucidum sterol is urgently needed.
The extraction and separation method of ganoderma triterpene and sterol mainly includes organic solvent extraction method, macroporous resin extraction method and ultrasonic treatment method. Chinese patent CN1629179A adopts ethanol reflux extraction and then alkalization to obtain the ganoderma lucidum sterol compounds, chinese patent CN1560072 adopts supercritical extraction and separation to obtain the ganoderma lucidum triterpenes and ganoderma lucidum polysaccharides, and Chinese patent CN101530436A adopts technologies such as ultrasonic wave, enzyme extraction and separation to obtain the ganoderma lucidum triterpenes and ganoderma lucidum polysaccharides. At present, relevant patents for simultaneously extracting and separating ganoderma triterpene and ganoderma sterol components are temporarily absent, and high-pressure supercritical extraction is adopted in literature reportsCO 2 The ganoderma triterpene and the sterol are extracted, but the obtained extract is a mixture, the ganoderma triterpene and the ganoderma sterol cannot be separated, and the high-pressure supercritical CO2 extraction method has the defects of large equipment investment, high operation difficulty and high production cost.
Disclosure of Invention
The invention aims to provide a simple and rapid method for simultaneously extracting and separating triterpene and sterol components in a ganoderma alcohol extract aiming at the defects of high separation difficulty, complex process, high production cost and the like of ganoderma triterpene and sterol in the prior art.
In order to solve the technical problem, the invention adopts the following technical scheme:
a method for rapidly separating triterpene and sterol from Ganoderma alcohol extract comprises loading polyamide resin into column, and eluting Ganoderma alcohol extract with water solution of organic solvent to separate triterpene and sterol.
Preferably, the organic solvent comprises one or more of ethanol, methanol and acetone.
Preferably, the polyamide resin column packing comprises the following steps:
A. pre-soaking: soaking the polyamide resin raw material in the organic solvent, and removing bubbles by ultrasonic treatment or stirring, wherein the soaking time is more than 10 hours;
B. column assembling: filling the polyamide resin soaked in the step A into a column, and washing the column with water until the column is odorless to obtain a polyamide resin column;
C. sampling: and B, mixing the ganoderma alcohol extract and the polyamide resin soaked in the step A according to a mass ratio of 1:1-2, evaporating to dryness, and filling the mixture into a polyamide resin column to finish sample loading, wherein the sample loading amount is not more than 2.5g/100ml.
The purpose of the pre-soaking is to allow the filler to swell sufficiently in the solvent and to dissolve impurities (waxes) on the surface of the filler. The soaking time is not completely fixed, and is generally estimated according to the amount of the filler, and the use effect can be achieved by soaking for more than 10 hours. And wet column packing and water washing are adopted to remove the organic solvent and impurities dissolved in the organic solvent, so that the separation effect of the polyamide resin is prevented from being influenced. And (3) adopting a dry method for loading, uniformly stirring the ganoderma alcohol extract and the polyamide resin, evaporating the solvent to dryness, and filling into a polyamide resin column. Thus, the method combining wet column packing and dry sample loading can ensure the quality of the filler and the sample loading quality to the maximum extent and ensure the subsequent separation of triterpene and sterol.
Preferably, the elution comprises the steps of:
a. washing the column with 40-60% of aqueous solution of the organic solvent for 3-5 times of the column volume to obtain an eluent of the triterpene;
b. and (3) washing the column by using 80-90% of aqueous solution of the organic solvent, and washing by 3-5 times of the column volume to obtain the sterol eluent.
Preferably, in step a, the elution solution of the triterpene is evaporated to dryness to obtain the triterpene.
Preferably, in step b, the eluate of the sterol is evaporated to dryness to obtain the sterol.
Preferably, the evaporation to dryness is one or more of reduced pressure drying, spray drying and freeze drying.
Preferably, the ratio of the column diameter of the polyamide resin packed column to the height of the packed column is 1:5 to 20.
Preferably, the polyamide resin is 100-400 mesh.
A triterpene and sterol product obtained by the method for rapidly separating triterpene and sterol from ganoderic alcohol extract is provided.
The ganoderma triterpene and the sterol are series substances rather than single compounds, which also increases the separation difficulty of the ganoderma triterpene and the sterol. The application of the invention can rapidly separate the two types of substances and provide technical support for independently researching and applying the two types of substances.
Compared with the prior art, the implementation of the invention has the following beneficial effects:
1. aiming at the defects of the prior art, the method has the advantages that the ganoderma triterpene and sterol components are quickly separated, the operation is very simple and convenient, and the separation accuracy is high;
2. the method has the advantages of complete separation, good reproducibility, low cost, high safety, and suitability for industrial production.
Drawings
FIG. 1 is a graph showing the extraction and separation results of ganoderma triterpene and sterol in example 1 of the present invention;
FIG. 2 is a graph showing the extraction and separation results of ganoderma triterpene and sterol in example 2 of the present invention;
FIG. 3 is a graph showing the extraction and separation results of ganoderma triterpene and sterol in example 3 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail with reference to the accompanying drawings so that those skilled in the art can better understand the present invention and can implement the present invention, but the present invention is not limited by the illustrated examples. Modifications or substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and scope of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.
Example 1
S1, taking a proper amount of polyamide resin (100-200 meshes), adding ethanol for immersion, performing ultrasonic treatment for 10min to remove bubbles, stirring, and standing for 10h;
s2, column filling: selecting a glass chromatographic column with the diameter of 2.6cm, filling the column until the column height is 20cm and the column diameter ratio is 1:7, and eluting the column with deionized water until no alcohol smell exists;
s3, taking 1.0g of alcohol extract of the ganoderma lucidum fruiting body, mixing the alcohol extract with the polyamide resin soaked in the step S1 according to the mass ratio of 1:1, evaporating the ethanol in an evaporation dish to dryness, and then loading the sample;
s4, washing the column by 50% ethanol for 3 times of column volume to obtain an eluent A; then washing the column with 90% ethanol by 3 times of the column volume to obtain eluent B;
s5, carrying out chromatographic detection on the eluent A and the eluent B, wherein the result is shown in figure 1, and it can be seen from figure 1 that the ganoderma triterpenoid component is completely eluted in the eluent A and the ganoderma sterol component is completely eluted in the eluent B; performing rotary evaporation on the eluent A under reduced pressure, and drying to obtain ganoderma triterpene component; carrying out reduced pressure rotary evaporation and drying on the eluent B to obtain a ganoderic sterol component;
s6, taking a proper amount of triterpene components to react with vanillin-glacial acetic acid, wherein the solution is purple and reacts with Liebermann-Burchard reagent without sterol color reaction; taking a proper amount of ganoderma lucidum sterol components to react with Liebermann-Burchard reagent, wherein the solution is green and reacts with vanillin-glacial acetic acid without triterpene color reaction; the obtained ganoderma triterpene and sterol components are completely separated and have no cross.
Example 2
S1, taking a proper amount of polyamide resin (100-200 meshes), adding ethanol for immersion, performing ultrasonic treatment for 10min to remove bubbles, stirring, and standing for 15h;
s2, column filling: selecting a glass chromatographic column with the diameter of 5.5cm, filling the glass chromatographic column until the column height is 30cm and the column diameter ratio is 1:5, and eluting the glass chromatographic column with deionized water until no alcohol smell exists;
s3, taking 5.0g of alcohol extract of the ganoderma lucidum fruiting body, mixing the alcohol extract with the polyamide resin soaked in the step S1 according to a mass ratio of 1.5, evaporating ethanol in an evaporation dish, and then loading the sample;
s4, washing the column by 55% ethanol, and washing by 5 times of the column volume to obtain an eluent A; then, washing the column by using 85% ethanol, and washing the column by 5 times of the column volume to obtain eluent B;
s5, carrying out chromatographic detection on the eluent A and the eluent B, wherein the result is shown in figure 2, and it can be seen from figure 2 that the ganoderma triterpenoid component is completely eluted in the eluent A and the ganoderma sterol component is completely eluted in the eluent B; performing rotary evaporation on the eluent A under reduced pressure, and drying to obtain ganoderma triterpene component; carrying out reduced pressure rotary evaporation and drying on the eluent B to obtain a ganoderic sterol component;
s6, taking a proper amount of triterpene components to react with vanillin-glacial acetic acid, enabling a solution to be purple, and reacting with a Liebermann-Burchard reagent without sterol color reaction; taking a proper amount of ganoderma lucidum sterol components to react with Liebermann-Burchard reagent, wherein the solution is green and reacts with vanillin-glacial acetic acid without triterpene color reaction; the obtained ganoderma triterpene and sterol components are completely separated and have no cross.
Example 3
S1, taking a proper amount of polyamide resin (100-200 meshes), adding ethanol for immersion, stirring to remove bubbles, and standing for 12 hours after stirring;
s2, column filling: selecting a glass chromatographic column with the diameter of 6cm, filling the column until the column height is 36cm and the column diameter ratio is 1:6, and eluting the column with deionized water until no alcohol smell exists;
s3, taking 10.0g of alcohol extract of the ganoderma lucidum fruiting body, mixing the alcohol extract with the polyamide resin soaked in the step S1 according to the mass ratio of 1:1, evaporating the ethanol in an evaporation dish to dryness, and then loading the sample;
s4, washing the column by 45% ethanol for 3 times of column volume to obtain an eluent A; then washing the column with 80% ethanol for 3 times of column volume to obtain eluent B;
s5, carrying out chromatographic detection on the eluent A and the eluent B, wherein the result is shown in figure 3, and it can be seen from figure 3 that the ganoderma triterpenoid component is completely eluted in the eluent A and the ganoderma sterol component is completely eluted in the eluent B; performing rotary evaporation on the eluent A under reduced pressure, and drying to obtain ganoderma triterpene component; carrying out reduced pressure rotary evaporation and drying on the eluent B to obtain a ganoderic sterol component;
s6, taking a proper amount of triterpene components to react with vanillin-glacial acetic acid, wherein the solution is purple and reacts with Liebermann-Burchard reagent without sterol color reaction; taking a proper amount of ganoderma lucidum sterol components to react with Liebermann-Burchard reagent, wherein the solution is green and reacts with vanillin-glacial acetic acid without triterpene color reaction; the obtained ganoderma triterpene and sterol components are completely separated and have no cross.
Example 4:
s1, taking a proper amount of polyamide resin (100-200 meshes), adding ethanol for immersion, stirring to remove bubbles, and standing for 12 hours after stirring;
s2, column assembling: selecting a glass chromatographic column with the diameter of 10cm, filling the glass chromatographic column until the column height is 70cm and the column diameter ratio is 1:7, and eluting the glass chromatographic column with deionized water until no alcohol smell exists;
s3, taking 50.0g of alcohol extract of the ganoderma lucidum fruiting body, mixing the alcohol extract with the polyamide resin soaked in the step S1 according to the mass ratio of 1:1, evaporating the ethanol in an evaporation dish to dryness, and then loading the sample;
s4, washing the column by 50% ethanol for 3 times of column volume to obtain an eluent A; then washing the column with 90% ethanol for 3 times of column volume to obtain eluent B;
s5, carrying out reduced pressure rotary steaming and drying on the eluent A to obtain a ganoderma triterpene component; the eluent B is decompressed, steamed and dried to obtain the ganoderic sterol component;
s6, taking a proper amount of triterpene components to react with vanillin-glacial acetic acid, wherein the solution is purple and reacts with Liebermann-Burchard reagent without sterol color reaction; taking a proper amount of ganoderma lucidum sterol components to react with Liebermann-Burchard reagent, wherein the solution is green and reacts with vanillin-glacial acetic acid without triterpene color reaction; the obtained ganoderma triterpene and sterol components are completely separated and have no cross.
Example 5
S1, taking a proper amount of polyamide resin (100-200 meshes), adding methanol for immersion, performing ultrasonic treatment for 10min to remove bubbles, stirring, and standing for 14h;
s2, column assembling: selecting a glass chromatographic column with the diameter of 5.5cm, filling the glass chromatographic column until the column height is 35cm and the column diameter ratio is 1:6, and eluting the glass chromatographic column with deionized water until no alcohol smell exists;
s3, taking 6.0g of alcohol extract of the ganoderma lucidum fruiting body, mixing the alcohol extract with the polyamide resin soaked in the step S1 according to the mass ratio of 1:1, evaporating methanol in an evaporation dish to dryness, and then loading the sample;
s4, washing the column by 50% methanol for 4 times of the column volume to obtain an eluent A; then washing the column with 90% methanol by 4 times of the column volume to obtain eluent B;
s5, carrying out reduced pressure rotary steaming and drying on the eluent A to obtain a ganoderma triterpene component; carrying out reduced pressure rotary evaporation and drying on the eluent B to obtain a ganoderic sterol component;
s6, taking a proper amount of triterpene components to react with vanillin-glacial acetic acid, wherein the solution is purple and reacts with Liebermann-Burchard reagent without sterol color reaction; taking a proper amount of ganoderma lucidum sterol components to react with Liebermann-Burchard reagent, wherein the solution is green and reacts with vanillin-glacial acetic acid without triterpene color reaction; the obtained ganoderma triterpene and sterol components are completely separated and have no cross.
Example 6
S1, taking a proper amount of polyamide resin (100-200 meshes), adding methanol for immersion, performing ultrasonic treatment for 10min to remove bubbles, stirring, and standing for 12h;
s2, column filling: selecting a glass chromatographic column with the diameter of 2.8cm, filling the glass chromatographic column until the column height is 18cm and the column diameter ratio is 1:6, and eluting the glass chromatographic column with deionized water until no alcohol smell exists;
s3, taking 2.0g of alcohol extract of the ganoderma lucidum fruiting body, mixing the alcohol extract with the polyamide resin soaked in the step S1 according to 1:1, evaporating methanol in an evaporation dish to dryness, and then loading the sample;
s4, washing the column by 40% methanol, and washing by 3 times of column volume to obtain an eluent A; then, washing the column by using 80% methanol for 3 times of the column volume to obtain eluent B;
s5, carrying out reduced pressure rotary steaming and drying on the eluent A to obtain a ganoderma triterpene component; carrying out reduced pressure rotary evaporation and drying on the eluent B to obtain a ganoderic sterol component;
s6, taking a proper amount of triterpene components to react with vanillin-glacial acetic acid, wherein the solution is purple and reacts with Liebermann-Burchard reagent without sterol color reaction; taking a proper amount of ganoderma lucidum sterol components to react with Liebermann-Burchard reagent, wherein the solution is green and reacts with vanillin-glacial acetic acid without triterpene color reaction; the obtained ganoderma triterpene and sterol components are completely separated and have no cross.
Example 7
S1, taking a proper amount of polyamide resin (200-400 meshes), adding acetone for immersion, performing ultrasonic treatment for 10min to remove bubbles, stirring, and standing for 10h;
s2, column filling: selecting a glass chromatographic column with the diameter of 2.8cm, filling the glass chromatographic column until the column height is 18cm and the column diameter ratio is 1:6, and eluting the glass chromatographic column with deionized water until the column is odorless;
s3, taking 3.0g of alcohol extract of the ganoderma lucidum fruiting body, mixing the alcohol extract with the polyamide resin soaked in the step S1 according to the mass ratio of 1:1, evaporating acetone in an evaporation dish to dryness, and then loading the sample;
s4, washing the column by using 60% acetone, and washing by 3 times of the column volume to obtain an eluent A; then washing the column with 90% acetone by 3 times of the column volume to obtain eluent B;
s5, carrying out reduced pressure rotary steaming and drying on the eluent A to obtain a ganoderma triterpene component; carrying out reduced pressure rotary evaporation and drying on the eluent B to obtain a ganoderic sterol component;
s6, taking a proper amount of triterpene components to react with vanillin-glacial acetic acid, wherein the solution is purple and reacts with Liebermann-Burchard reagent without sterol color reaction; taking a proper amount of ganoderma lucidum sterol components to react with Liebermann-Burchard reagent, wherein the solution is green and reacts with vanillin-glacial acetic acid without triterpene color reaction; the obtained ganoderma triterpene and sterol components are completely separated and have no cross.
Example 8
S1, taking a proper amount of polyamide resin (100-200 meshes), adding acetone for immersion, stirring to remove bubbles, and standing for 12 hours after stirring;
s2, column filling: selecting a glass chromatographic column with the diameter of 3cm, filling the column until the column height is 21cm and the column diameter ratio is 1:7, and eluting the column with deionized water until the column is odorless;
s3, taking 4.0g of alcohol extract of the ganoderma lucidum fruiting body, mixing the sample with the soaked polyamide resin in the step S1 according to the mass ratio of 1:2, evaporating acetone in an evaporation dish, and then loading the sample;
s4, washing the column by 50% acetone for 3 times of the column volume to obtain an eluent A; then washing the column with 90% acetone by 3 times of the column volume to obtain eluent B;
s5, carrying out reduced pressure rotary steaming and drying on the eluent A to obtain a ganoderma triterpene component; carrying out reduced pressure rotary evaporation and drying on the eluent B to obtain a ganoderic sterol component;
s6, taking a proper amount of triterpene components to react with vanillin-glacial acetic acid, wherein the solution is purple and reacts with Liebermann-Burchard reagent without sterol color reaction; taking a proper amount of ganoderma lucidum sterol components to react with Liebermann-Burchard reagent, wherein the solution is green and reacts with vanillin-glacial acetic acid without triterpene color reaction; the obtained triterpene and sterol components of the ganoderma lucidum are completely separated and have no cross.
The above disclosure is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the scope of the present invention, therefore, the present invention is not limited by the appended claims.

Claims (8)

1. A method for rapidly separating triterpene and sterol from alcohol extract of Ganoderma comprises loading polyamide resin into column, eluting the alcohol extract of Ganoderma with water solution of organic solvent to separate triterpene and sterol; the organic solvent comprises one or more of ethanol, methanol and acetone;
the elution comprises the following steps:
a. eluting the column by using 40-60% of aqueous solution of the organic solvent, and eluting 3~5 times of the column volume to obtain an eluent of the triterpene;
b. and (3) washing the column by using 80-90% of aqueous solution of the organic solvent, and washing 3~5 times of the column volume to obtain the sterol eluent.
2. The method for rapidly separating triterpene and sterol from ganoderic alcohol extract according to claim 1, wherein the step of loading polyamide resin into column comprises the steps of:
A. pre-soaking: soaking the polyamide resin raw material in the organic solvent, and removing bubbles by ultrasonic treatment or stirring, wherein the soaking time is more than 10 hours;
B. column assembling: filling the polyamide resin soaked in the step A into a column, and washing the column with water until the column is odorless to obtain a polyamide resin column;
C. loading: and (B) mixing the ganoderma alcohol extract and the polyamide resin soaked in the step (A) according to a mass ratio of 1-2, evaporating to dryness, and filling the mixture into the polyamide resin column to finish sample loading, wherein the sample loading amount is not more than 2.5g/100ml.
3. The method for rapidly separating triterpene and sterol from ganoderic acid alcohol extract according to claim 1, wherein in step a, the triterpene is obtained by evaporating the elution solution of triterpene.
4. The method for rapidly separating triterpene and sterol from alcohol extract of Ganoderma lucidum as claimed in claim 1, wherein in step b, the eluent of sterol is evaporated to dryness to obtain sterol.
5. The method for rapidly separating triterpene and sterol from ganoderic alcohol extract according to claim 3 or 4, wherein the evaporation is one or more of reduced pressure drying, spray drying and freeze drying.
6. The method for rapidly separating triterpene and sterol in ganoderma lucidum alcohol extract according to claim 1, wherein the ratio of the column diameter to the packed column height of the polyamide resin is 1 to 5-20.
7. The method for rapidly separating triterpene and sterol from alcohol extract of Ganoderma lucidum as claimed in claim 1, wherein the polyamide resin is 100-400 mesh.
8. A triterpene and sterol product obtained by the method for rapidly separating triterpene and sterol from alcohol extract of Ganoderma lucidum as claimed in claim 1.
CN202011525201.5A 2020-12-22 2020-12-22 Method for rapidly separating triterpene and sterol in ganoderma lucidum alcohol extract Active CN112603932B (en)

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