CN111494442A - Method for improving extraction yield of total saponins in akebia trifoliata peels - Google Patents

Method for improving extraction yield of total saponins in akebia trifoliata peels Download PDF

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CN111494442A
CN111494442A CN202010487718.3A CN202010487718A CN111494442A CN 111494442 A CN111494442 A CN 111494442A CN 202010487718 A CN202010487718 A CN 202010487718A CN 111494442 A CN111494442 A CN 111494442A
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akebia trifoliata
peels
akebia
enzymolysis
total saponins
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钱卫东
张家宁
孙照欢
杨敏
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Shaanxi University of Science and Technology
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    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K36/18Magnoliophyta (angiosperms)
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    • 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
    • A61K2236/10Preparation or pretreatment of starting material
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    • A61K2236/30Extraction of the material
    • A61K2236/33Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • 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
    • A61K2236/51Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying

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Abstract

A method for improving the extraction of total saponins in akebia trifoliata peels comprises the following steps: mixing the akebia trifoliata peel powder with pectinase, cellulase, amylase and polyether serving as an enzyme activator, and placing the mixture into a reaction tank for enzymolysis to obtain akebia trifoliata peel powder enzymatic hydrolysate; adding 70% methanol into the akebia trifoliata peel powder enzymolysis liquid, and performing ultrasonic extraction to obtain an extracting solution and filter residues; adsorbing the extract by macroporous resin, washing, eluting, and collecting Akebia trifoliata peel powder eluate; adjusting the pH value of the eluent, and concentrating and recycling the eluent; freeze drying to obtain the total saponin extract in the pericarp of akebia trifoliata. Compared with the traditional method for extracting the akebia trifoliata peels by alcohol, the akebia trifoliata peels are prepared into akebia trifoliata peel powder, and are subjected to enzymolysis by pectinase, cellulase, amylase and polyether serving as an enzyme activator to obtain akebia trifoliata enzymatic hydrolysate for extraction, so that the extraction yield of the total saponins as the active ingredients in the akebia trifoliata peels is effectively improved.

Description

Method for improving extraction yield of total saponins in akebia trifoliata peels
Technical Field
The invention belongs to the technical field of traditional Chinese medicine extraction, and particularly relates to a method for improving the extraction yield of total saponins in akebia trifoliata peels.
Background
In recent years, the production and extraction methods of Chinese herbal medicines are developed and mechanized to a certain extent, and research papers on the extraction methods of Chinese herbal medicines are also provided, however, the traditional Chinese herbal medicines are considered to have more impurities and have unstable and lower active ingredient content, and can not be in contact with modern medicine, thereby restricting the development of modern Chinese medicines.
The akebia trifoliata peels have complex chemical components and various structures, mainly comprise triterpenes and glycosides thereof, and triterpenoid saponin compounds are separated from akebia trifoliata peels at present, wherein the aglycone types comprise hederagenin, norhederagenin, oleanane sapogenin, noroleanane sapogenin, argatropurpurin, ursolic sapogenin and the like. Has antibacterial, antiinflammatory, tumor growth inhibiting, hepatoprotective, enzyme reducing, and immunity improving effects. The traditional method for extracting the effective components of the akebia trifoliata peels is an alcohol extraction method, and the extraction rate of the total saponins in the akebia trifoliata peels is low. The extraction of effective substances from traditional plants is an important unit operation and is the starting point in the research of most traditional Chinese herbal medicines. The quality of the extraction method and the achievement is directly related to the difficulty of the subsequent experiment progress. The extraction method can be regarded as an important link of modern Chinese herbal medicine experiments. Therefore, it is necessary to research and optimize the extraction method of the effective substances of the traditional Chinese herbal medicines.
Disclosure of Invention
The invention aims to provide a method capable of effectively extracting the total saponin extraction yield from akebia trifoliata peels.
The invention is realized by the following technical scheme:
q1: cleaning caulis Akebiae pericarp, sun drying, slicing, pulverizing, and sieving with 40 mesh sieve to obtain refined caulis Akebiae pericarp powder;
q2: adding 0.02-0.05 g of pectinase, 0.02-0.05 g of cellulase and 0.02-0.05 g of amylase into each kilogram of akebia trifoliata peel powder respectively, adding an enzyme activator polyether into the mixture according to the mass ratio of 0.3-0.5% of pectinase, mixed enzyme of cellulase and amylase and the enzyme activator polyether, uniformly mixing the materials, and carrying out enzymolysis in a reaction tank to obtain akebia trifoliata peel enzymatic hydrolysate;
q3: adding 70% methanol into the akebia trifoliata peel enzymatic hydrolysate according to the volume ratio of 1: 6-10, fully stirring and extracting, separating an extracting solution and filter residues, and repeating for 2-3 times;
q4: sequentially adsorbing, washing and eluting the extracting solution, and collecting akebia trifoliata peel powder eluent; concentrating the eluate, and freeze drying to obtain total saponin extract.
And the slicing and crushing in the step Q1 is to cut the akebia trifoliata peels into 3-5 mm slices and then put the akebia trifoliata peels into a traditional Chinese medicine crusher for crushing.
And the enzymolysis temperature of the step Q2 is 35-55 ℃, enzymolysis is carried out in the reaction tank for 2-3 h, then the pressure in the reaction tank is restored to be in a normal pressure state, and the akebia trifoliata peel enzymolysis liquid is obtained.
The enzymolysis in the steps is carried out for 0.5h at 35 ℃, 1h at 45 ℃ and 1h at 55 ℃ in sequence.
The pressure in the reaction tank in the enzymolysis process is 0.4-0.6 Mpa, and the pH value is 4-5.
The temperature of the methanol added in the step Q3 is 70-90 ℃.
Adsorbing and enriching by using a macroporous resin adsorption method in the step Q4, washing by using 40% ethanol and purified water, and eluting by using 95% ethanol solution to realize concentration to obtain an eluent; adjusting the pH value of the eluent to 2-4, and concentrating and recovering the eluent at 40-60 ℃ for 60-90 min to obtain a concentrated solution.
Compared with the prior art, the invention has the following beneficial technical effects:
compared with the traditional method for extracting the total saponins of the active ingredients in the akebia trifoliata peel powder, the method provided by the invention has the advantages that the akebia trifoliata peel is prepared into the akebia trifoliata peel powder, and the akebia trifoliata peel enzymatic hydrolysate is obtained through the enzymolysis of pectinase, cellulase, amylase and an enzyme activator polyether and then extracted, so that the extraction yield of the total saponins of the active ingredients in the akebia trifoliata peel of the traditional Chinese medicine is effectively improved.
Detailed Description
Specific examples are given below.
Example 1:
q1: cleaning caulis Akebiae pericarp, sun drying, cutting into 3mm pieces, pulverizing in a traditional Chinese medicine pulverizer, sieving with 40 mesh micro powder vibration sieve to obtain refined caulis Akebiae pericarp powder;
q2: carrying out enzymolysis reaction on akebia trifoliata peel powder; the method specifically comprises the following steps:
q21: adding 0.05g of pectinase, 0.05g of cellulase and 0.05g of amylase into each kilogram of akebia trifoliata peel powder respectively, and then adding an enzyme activator polyether into the mixture according to the mass ratio of 0.5 percent of pectinase, mixed enzyme of cellulase and amylase and the enzyme activator polyether to uniformly mix to obtain an enzyme mixture;
q22: uniformly paving the enzyme mixture in a reaction tank, and closing the reaction tank;
q23: adjusting the temperature in the reaction tank to 35 ℃, the pressure to 0.5MPa, the pH value to 4.5, and carrying out enzymolysis for 0.5 h; adjusting the temperature in the reaction tank to 45 ℃, the pressure to 0.5MPa, the pH value to 4.5, and carrying out enzymolysis for 1 h; adjusting the temperature in the reaction tank to 55 ℃, the pressure to 0.5MPa, the pH value to 4.5, and carrying out enzymolysis for 1 h; then recovering the pressure in the reaction tank to normal pressure to obtain akebia trifoliata peel enzymatic hydrolysate;
q3: adding 70% methanol at 70 deg.C into caulis Akebiae hydrolysate at a volume ratio of 1:8, stirring and extracting, separating extractive solution and residue, and repeating for 3 times;
q4: adsorbing and enriching the eluent by using a macroporous resin adsorption method, washing the eluent by using ethanol with the volume concentration of 40% and purified water, and eluting the eluent by using ethanol solution with the volume concentration of 95% to realize concentration to obtain eluent; adjusting pH of the eluate to 3, concentrating at 50 deg.C for 75min, recovering, and freeze drying to obtain total saponin extract.
The extraction rate of the total saponins in the pericarp of akebia trifoliata is calculated by the following formula:
extraction yield/% -% m1/m0×100
In the formula: m is1The mass/g of the akebia trifoliata peel total saponins is as follows: m is0Is the mass/g of the raw material.
The total saponin content was measured with HP L C, and the results showed that the total saponin extraction rate was 0.64%.
Example 2:
q1: cleaning caulis Akebiae pericarp, sun drying, cutting into 4mm pieces, pulverizing in a traditional Chinese medicine pulverizer, sieving with 40 mesh micro powder vibration sieve to obtain refined caulis Akebiae pericarp powder;
q2: carrying out enzymolysis reaction on akebia trifoliata peel powder; the method specifically comprises the following steps:
q21: adding 0.02g of pectinase, 0.02g of cellulase and 0.02g of amylase into each kilogram of akebia trifoliata peel powder respectively, and then adding an enzyme activator polyether into the mixture according to the mass ratio of 0.4 percent of pectinase, mixed enzyme of cellulase and amylase and the enzyme activator polyether to uniformly mix to obtain an enzyme mixture;
q22: uniformly paving the enzyme mixture in a reaction tank, and closing the reaction tank;
q23: adjusting the temperature in the reaction tank to 35 ℃, the pressure to 0.4MPa, the pH value to 4, and carrying out enzymolysis for 0.5 h; adjusting the temperature in the reaction tank to 45 ℃, the pressure to 0.4MPa, the pH value to 4, and carrying out enzymolysis for 1 h; adjusting the temperature in the reaction tank to 55 ℃, the pressure to 0.4MPa, the pH value to 4, and carrying out enzymolysis for 1 h; then recovering the pressure in the reaction tank to normal pressure to obtain akebia trifoliata peel enzymatic hydrolysate;
q3: adding 70% methanol with the concentration of 80 ℃ into the akebia trifoliata peel enzymolysis liquid according to the volume ratio of 1:6, fully stirring and extracting, separating the extracting solution and filter residue, and repeating for 2 times;
q4: adsorbing and enriching the eluent by using a macroporous resin adsorption method, washing the eluent by using ethanol with the volume concentration of 40% and purified water, and eluting the eluent by using ethanol solution with the volume concentration of 95% to realize concentration to obtain eluent; adjusting pH of the eluate to 2, concentrating at 60 deg.C, recovering for 60min to obtain concentrated solution, and freeze drying to obtain total saponin extract.
The extraction rate of the total saponins in the pericarp of akebia trifoliata is calculated by the following formula:
extraction yield/% -% m1/m0×100
In the formula: m is1The mass/g of the akebia trifoliata peel total saponins is as follows: m is0Is the mass/g of the raw material.
The total saponin content was analyzed by HP L C, and the results showed that the total saponin extraction rate was 0.57%.
Example 3:
q1: cleaning caulis Akebiae pericarp, sun drying, cutting into 5mm pieces, pulverizing in a traditional Chinese medicine pulverizer, sieving with 40 mesh micro powder vibration sieve to obtain refined caulis Akebiae pericarp powder;
q2: carrying out enzymolysis reaction on akebia trifoliata peel powder; the method specifically comprises the following steps:
q21: adding 0.03g of pectinase, 0.03g of cellulase and 0.03g of amylase into each kilogram of akebia trifoliata peel powder respectively, and then adding an enzyme activator polyether into the mixture according to the mass ratio of 0.3 percent of pectinase, mixed enzyme of cellulase and amylase and the enzyme activator polyether to uniformly mix to obtain an enzyme mixture;
q22: uniformly paving the enzyme mixture in a reaction tank, and closing the reaction tank;
q23: adjusting the temperature in the reaction tank to 35 ℃, the pressure to 0.6MPa, the pH value to 5, and carrying out enzymolysis for 0.5 h; adjusting the temperature in the reaction tank to 45 ℃, the pressure to 0.6MPa, the pH value to 5, and carrying out enzymolysis for 1 h; adjusting the temperature in the reaction tank to 55 ℃, the pressure to 0.6MPa, the pH value to 5, and carrying out enzymolysis for 1 h; then recovering the pressure in the reaction tank to normal pressure to obtain akebia trifoliata peel enzymatic hydrolysate;
q3: adding 70% methanol at 90 deg.C into caulis Akebiae peel enzymolysis solution at volume ratio of 1:10, stirring and extracting, separating extractive solution and residue, and repeating for 3 times;
q4: adsorbing and enriching the eluent by using a macroporous resin adsorption method, washing the eluent by using ethanol with the volume concentration of 40% and purified water, and eluting the eluent by using ethanol solution with the volume concentration of 95% to realize concentration to obtain eluent; adjusting pH of the eluate to 4, concentrating at 40 deg.C, recovering for 90min to obtain concentrated solution, and freeze drying to obtain total saponin extract.
The extraction rate of the total saponins in the pericarp of akebia trifoliata is calculated by the following formula:
extraction yield/% -% m1/m0×100
In the formula: m is1The mass/g of the akebia trifoliata peel total saponins is as follows: m is0Is the mass/g of the raw material.
The total saponin content was measured with HP L C, and the results showed that the total saponin extraction yield was 0.53%.

Claims (7)

1. A method for improving the extraction yield of total saponins in akebia trifoliata peels is characterized by comprising the following steps:
q1: cleaning caulis Akebiae pericarp, sun drying, slicing, pulverizing, and sieving with 40 mesh sieve to obtain refined caulis Akebiae pericarp powder;
q2: adding 0.02-0.05 g of pectinase, 0.02-0.05 g of cellulase and 0.02-0.05 g of amylase into each kilogram of akebia trifoliata peel powder respectively, adding an enzyme activator polyether into the mixture according to the mass ratio of 0.3-0.5% of pectinase, mixed enzyme of cellulase and amylase and the enzyme activator polyether, uniformly mixing the materials, and carrying out enzymolysis in a reaction tank to obtain akebia trifoliata peel enzymatic hydrolysate;
q3: adding 70% methanol into the akebia trifoliata peel enzymatic hydrolysate according to the volume ratio of 1: 6-10, fully stirring and extracting, separating an extracting solution and filter residues, and repeating for 2-3 times;
q4: sequentially adsorbing, washing and eluting the extracting solution, and collecting akebia trifoliata peel powder eluent; concentrating the eluate, and freeze drying to obtain total saponin extract.
2. The method for improving the extraction yield of the total saponins in the akebia trifoliata peels as claimed in claim 1, wherein the slicing and crushing in the step Q1 are carried out by cutting the akebia trifoliata peels into 3-5 mm slices and then crushing the akebia trifoliata peels in a traditional Chinese medicine crusher.
3. The method for improving the extraction yield of the total saponins in the akebia trifoliata peels as claimed in claim 1, wherein the enzymolysis temperature in the step Q2 is 35-55 ℃, the enzymolysis is carried out in a reaction tank for 2-3 h, and then the pressure in the reaction tank is restored to be in a normal pressure state, so that akebia trifoliata peel enzymolysis liquid is obtained.
4. The method for improving the extraction yield of the total saponins in the akebia trifoliata peels as claimed in claim 3, wherein the enzymolysis in the step is sequentially carried out at 35 ℃ for 0.5h, 45 ℃ for 1h and 55 ℃ for 1 h.
5. The method for improving the extraction yield of the total saponins in the akebia trifoliata peels as claimed in claim 3, wherein the pressure in the reaction tank in the enzymolysis process is 0.4-0.6 Mpa, and the pH value is 4-5.
6. The method for improving the extraction rate of the total saponins in the pericarp of akebia trifoliata as claimed in claim 1, wherein the temperature of the methanol added in the step Q3 is 70-90 ℃.
7. The method for improving the extraction yield of the total saponins in the pericarp of akebia trifoliata as claimed in claim 1, wherein the step Q4 is implemented by using a macroporous resin adsorption method for adsorption and enrichment, washing with 40% ethanol and purified water, and then eluting with 95% ethanol solution for concentration to obtain an eluent; adjusting the pH value of the eluent to 2-4, and concentrating and recovering the eluent at 40-60 ℃ for 60-90 min to obtain a concentrated solution.
CN202010487718.3A 2020-06-02 2020-06-02 Method for improving extraction yield of total saponins in akebia trifoliata peels Pending CN111494442A (en)

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