CN112704074B - Method for extracting natural plant endogenous components - Google Patents
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Abstract
The invention discloses a method for extracting natural plant endogenous components, which comprises the following steps: s1, drying and crushing natural plants containing flavone and shikimic acid to obtain plant powder, adding water and enzyme into the plant powder for enzymolysis, and inactivating to obtain an enzymolysis product; s2, adding polyvinylpyrrolidone and butanediol into the enzymolysis product, carrying out ultrasonic treatment for 20-40min, then adding an ethanol water solution, heating, refluxing, extracting, and filtering to obtain a filtrate; s3, putting the filtrate into a chitosan adsorption column, when the pH of the effluent is 4-5, leaching the chitosan adsorption column by using an ethanol aqueous solution until the effluent is neutral, then eluting by using an ethanol aqueous solution containing sodium hydroxide with the pH of 9-10, and collecting the effluent; and S4, purifying, concentrating and drying the effluent to obtain the plant endogenous component. The invention provides a method for extracting endogenous ingredients of natural plants, which effectively improves the extraction rate of shikimic acid and flavone in the natural plants, and the obtained endogenous ingredients of the plants can be widely applied to agricultural production and have good economic benefit.
Description
Technical Field
The invention belongs to the technical field of plant extraction, and relates to a method for extracting natural plant endogenous components.
Background
The flavonoids generally refer to a basic parent nucleus, and are a series of compounds formed by connecting benzene rings (and rings) of phenyl chromone with pink groups through a central three-carbon atom, and have functional groups such as methoxy, common amino, methyl, isopentenyl and the like. According to the oxidation degree of the central three-carbon chain, the ring connecting position of 2 or 3 positions, the cyclic structure of the three-carbon chain and other characteristics, the main natural flavonoid compounds can be classified as follows: fifteen kinds of flavonoid, flavonol, anthocyanidin, flavonone, flavanonol, etc. The flavone has various effects, is a strong antioxidant, can effectively remove oxygen free radicals in vivo, has oxidation preventing capability more than ten times of that of vitamins, and can prevent cell degeneration, aging and cancer. The flavone also has remarkable physiological activities of improving blood circulation, reducing cholesterol, enhancing immunity, protecting central nervous system and cardiovascular and cerebrovascular systems, and the like. The flavonoids can promote photosynthesis of plants, are beneficial to growth and development of roots, stems and leaves of the plants, and play an important role in promoting growth, yield and quality formation of crops such as tea leaves, hot peppers, rice, tobacco leaves, eggplants, vegetables, grapes and oranges. The crops sprayed with flavonoids have good growth vigor, the plant height is obviously increased, and the cold resistance is also improved to a certain degree. In addition, the flavonoid substance has good sterilization, insecticidal and antiviral effects, can be used as a botanical pesticide component, and has good environmental and biological safety.
Shikimic acid and its derivatives are gradually recognized in the medical field because they exhibit various pharmacological activities such as antibacterial, anti-inflammatory, antiviral, antithrombotic, etc. It is also the basic material of medicine "duffy" for treating avian influenza virus patient. It has been reported that the content of shikimic acid in pine needles of pinus massoniana is 2-3%, and the content of shikimic acid in pine needles of tender leaves is the highest, and the content of shikimic acid in pine needles of cedus massoniana and tender branches is 3.93% and 1.66%, respectively. China has abundant pine resources, so the shikimic acid in the pine needles becomes a new main plant resource. Currently, the general direction of research on this is mainly the optimization of extraction processes, such as organic solvent extraction, steam gym, etc.; component identification and content determination: gas chromatography-mass spectrometry, high performance liquid chromatography; the pharmacological efficacy.
Therefore, improving the extraction rate of active ingredients such as flavonoid, shikimic acid and the like in natural plants is one of the hot points of research in the agricultural field, is beneficial to improving the economic value of the natural plants, has important significance for the application of the flavonoid, the shikimic acid and the like in agriculture and related fields, and has wide prospect.
Disclosure of Invention
Based on the background technology, the invention provides the method for extracting the natural plant endogenous components, the polyvinylpyrrolidone and the butanediol are used as the dissolution promoters and matched with the chitosan adsorption column, the extraction rate of shikimic acid and flavone in the natural plant and the utilization rate of the chitosan adsorption column are improved, the obtained plant endogenous substances can be used in the agricultural field, and the method has good biological safety and environmental friendliness, and the extraction process is simple and easy to operate, and is beneficial to industrialization.
The invention provides a method for extracting natural plant endogenous components, which comprises the following steps:
s1, drying and crushing natural plants containing flavone and shikimic acid to obtain plant powder, adding water and enzyme into the plant powder for enzymolysis, and inactivating to obtain an enzymolysis product;
s2, adding polyvinylpyrrolidone and butanediol into the enzymolysis product, performing ultrasonic treatment for 20-40min, and adding the mixture with a concentrated volume
Heating and refluxing ethanol water solution with the concentration of 50-80%, and filtering to obtain filtrate, wherein the volume ratio of the ethanol water solution to the enzymolysis product is 1-2:1-2;
s3, putting the filtrate into a chitosan adsorption column, when the pH of the effluent is 4-5, leaching the chitosan adsorption column by using an ethanol aqueous solution with the volume concentration of 20-30% until the effluent is neutral, then eluting by using an ethanol aqueous solution containing sodium hydroxide with the pH of 9-10, and collecting the effluent;
and S4, purifying, concentrating and drying the effluent to obtain the plant endogenous component.
Preferably, in step S1, the enzyme is one or a mixture of two or more of cellulase, hemicellulase, protease and pectinase.
Preferably, the enzymolysis temperature is 20-40 deg.C, and the enzymolysis time is 10-20min.
The enzymolysis treatment is to effectively destroy the cell wall of the natural plant, and is beneficial to the dissolution and extraction of the effective components.
Preferably, in step S1, the weight ratio of the plant powder to the water is 1:2-5.
Preferably, in step S1, the natural plant containing flavone and shikimic acid is one of ginkgo leaves, pine needles and star anise.
Preferably, in step S2, the weight-to-volume ratio g of the total amount of polyvinylpyrrolidone and butanediol to the enzymolysis product: mL is 0.2-0.5:1.
preferably, the weight ratio of polyvinylpyrrolidone to butanediol is 1-2:1-2.
Preferably, the weight ratio of polyvinylpyrrolidone to butanediol is 1:1.
preferably, in step S3, the sample loading flow rate is 5-6BV/h, and the elution flow rate is 3-4BV/h.
Preferably, in step S3, the preparation method of the chitosan adsorption column comprises: wetting chitosan with water, and loading the chitosan into a column to obtain the chitosan adsorption column.
Preferably, the chitosan is fibroin modified chitosan, and the specific modification method comprises the following steps: dissolving chitosan in acetic acid water solution, adding mixed acid consisting of concentrated nitric acid and concentrated phosphoric acid, then adding sodium nitrite, and reacting at 20-30 ℃ to obtain carboxylated chitosan; adding fibroin into the carboxylated chitosan, and reacting at 30-40 ℃ to obtain the modified chitosan.
In the present invention, the concentration of the concentrated nitric acid is 86% or more of nitric acid, and the concentration of the concentrated phosphoric acid is 83% or more of phosphorus.
Preferably, the weight volume ratio of the chitosan to the mixed acid is g: mL is 1:4-5.
Preferably, the volume ratio of concentrated nitric acid to concentrated phosphoric acid in the mixed acid is 2:1.
preferably, the amount of sodium nitrite added is 0.5-1% of the total mass of the mixed acid.
Preferably, the weight ratio of the carboxylated chitosan to the fibroin is 1-3:0.2-0.5.
Preferably, in step S4, purification is performed using a cationic resin column.
The invention carries out enzymolysis treatment on natural plants, is beneficial to and accelerates the dissolution of effective components in the natural plants, and adopts polyvinylpyrrolidone and butanediol to carry out ultrasonic treatment on plant powder before extraction, the polyvinylpyrrolidone and the butanediol are matched to increase the solubility of the effective components in the plant powder, especially the dissolution rate of shikimic acid and flavonoid is improved, and the extraction rate is greatly improved; the extraction efficiency of the subsequent extraction steps is improved. The chitosan adsorption column adopted in the separation stage selectively carboxylates the chitosan, and then the chitosan reacts with the fibroin to obtain the modified chitosan, the amino group contained in the chitosan and the amino group on the silk branched chain effectively promote the adsorption of the adsorption column on active ingredients such as plant endogenous ingredients flavone and shikimic acid, and meanwhile, the fiber structure of the fibroin effectively reduces the swelling property of the chitosan, so that the later-stage elution and separation of the active ingredients are facilitated.
The spatial structure of butanediol containing hydroxyl and matched with polyvinylpyrrolidone in the invention also has certain influence on the swelling performance of chitosan, has certain effect of relieving the swelling tendency of chitosan, further reduces the swelling performance of chitosan, avoids colloidal compounds formed by chitosan and acidic components in plants, and thus improves the yield and the reutilization rate of the adsorption column.
The invention improves the extraction effect of endogenous ingredients, especially flavonoid substances, in natural plants, and the obtained endogenous ingredients have high flavonoid content, can be used as culture solution, novel antibacterial and insecticidal fertilizer and feed additive, can be widely applied to agricultural production, and has good economic benefit.
Detailed Description
The invention is described in detail with reference to the following specific examples, which are intended to illustrate and not to limit the invention, and all modifications, equivalents, and the like that are based on the invention are intended to be included within the scope of the invention.
Example 1
A method for extracting natural plant endogenous components comprises the following steps:
s1, drying and crushing fresh ginkgo leaves to obtain ginkgo leaf powder, adding water, vitamin enzyme and protease into the ginkgo leaf powder for enzymolysis at the temperature of 30 ℃, and inactivating to obtain an enzymolysis product, wherein the weight ratio of the ginkgo leaf powder to the water is 1:2;
s2, adding polyvinylpyrrolidone and butanediol into the enzymolysis product, carrying out ultrasonic treatment for 20min, then adding an ethanol water solution with the volume concentration of 50%, heating, refluxing, extracting, and filtering to obtain a filtrate, wherein the volume ratio of the ethanol water solution to the enzymolysis product is 0.5:1, the weight volume ratio g of the total amount of polyvinylpyrrolidone and butanediol to the enzymolysis product: mL is 0.2:1, the weight ratio of polyvinylpyrrolidone to butanediol is 1:2;
s3, putting the filtrate into a chitosan adsorption column, when the pH of the effluent is 4, leaching the chitosan adsorption column by using an ethanol water solution with the volume concentration of 20% until the effluent is neutral, then eluting by using an ethanol water solution containing sodium hydroxide with the pH of 9, and collecting the effluent;
and S4, performing polarity exchange on the effluent liquid by adopting cation exchange resin to obtain a purified liquid, and concentrating and drying the purified liquid to obtain the ginkgo leaf extract.
Example 2
A method for extracting natural plant endogenous components comprises the following steps:
s1, drying and crushing fresh ginkgo leaves to obtain ginkgo leaf powder, adding 20ml of water, pectinase and cellulase into 10g of ginkgo leaf powder, performing enzymolysis for 20min at 20 ℃, and inactivating to obtain an enzymolysis product;
s2, adding 5g of polyvinylpyrrolidone and 5g of butanediol into the enzymolysis product, carrying out ultrasonic treatment for 40min, then adding 30mL of 80% ethanol water solution in volume concentration, heating, refluxing, extracting, and filtering to obtain a filtrate;
s3, putting the filtrate into a chitosan adsorption column, wherein the sample loading flow rate is 5BV/h, when the pH of the effluent is 4, leaching the chitosan adsorption column by using an ethanol water solution with the volume concentration of 20% until the effluent is neutral, then eluting by using an ethanol water solution containing sodium hydroxide with the pH of 9, wherein the elution flow rate is 3BV/h, and collecting the effluent;
s4, performing polarity exchange on the effluent liquid by adopting cation exchange resin to obtain a purified liquid, and concentrating and drying the purified liquid to obtain a ginkgo leaf extract;
wherein, the chitosan in the chitosan adsorption column is fibroin modified chitosan, and the specific modification method comprises the following steps: dissolving chitosan in acetic acid water solution, adding mixed acid consisting of concentrated nitric acid with the concentration of 90% and concentrated phosphoric acid with the concentration of 85%, and then adding sodium nitrite to react at 20 ℃ to obtain carboxylated chitosan; adding fibroin into the carboxylated chitosan, and reacting at 30 ℃ to obtain modified chitosan; the weight volume ratio of the chitosan to the mixed acid is g: mL is 1:4, concentrating the nitric acid and the concentrated acid in the mixed acid
Volume ratio of phosphoric acid 2:1, the addition amount of sodium nitrite is 0.5 percent of the total mass of the mixed acid, and the carboxylated chitosan and the fibroin
The weight ratio of (1): 0.2. example 3
A method for extracting natural plant endogenous components comprises the following steps:
s1, drying and crushing fresh pine needles to obtain pine needle powder, adding 50ml of water, protease and cellulase into 10g of pine needle powder, performing enzymolysis for 10min at 30 ℃, and inactivating to obtain an enzymolysis product;
s2, adding 8g of polyvinylpyrrolidone and 4g of butanediol into the enzymolysis product, carrying out ultrasonic treatment for 20min, then adding 60mL of 80% ethanol water solution with volume concentration, heating, refluxing, extracting, and filtering to obtain a filtrate;
s3, feeding the filtrate into a chitosan adsorption column, wherein the sample loading flow rate is 6BV/h, when the pH of the effluent is 5, leaching the chitosan adsorption column by using an ethanol aqueous solution with the volume concentration of 30% until the effluent is neutral, then eluting by using an ethanol aqueous solution containing sodium hydroxide with the pH of 10, wherein the elution flow rate is 4BV/h, and collecting the effluent;
s4, performing polarity exchange on the effluent liquid by adopting cation exchange resin to obtain purified liquid, and concentrating and drying the purified liquid to obtain a pine needle extract;
wherein chitosan in the chitosan adsorption column is fibroin-modified chitosan, and the specific modification method comprises the following steps: dissolving chitosan in acetic acid water solution, adding mixed acid consisting of concentrated nitric acid with the concentration of 90% and concentrated phosphoric acid with the concentration of 85%, and then adding sodium nitrite to react at 20 ℃ to obtain carboxylated chitosan; adding fibroin into the carboxylated chitosan, and reacting at 30 ℃ to obtain modified chitosan; the weight volume ratio of the chitosan to the mixed acid is g: mL is 1:5, the volume ratio of concentrated nitric acid to concentrated phosphoric acid in the resultant acid is 2:1, the addition amount of sodium nitrite is 1 percent of the total mass of the mixed acid, and the weight ratio of the carboxylated chitosan to the fibroin is 1:0.5.
example 4
A method for extracting natural plant endogenous components comprises the following steps:
s1, crushing star anise (dried) to obtain fennel powder, adding 30ml of water, hemicellulase and cellulase into 10g of fennel powder, performing enzymolysis for 10min at 30 ℃, and inactivating to obtain an enzymolysis product;
s2, adding 5g of polyvinylpyrrolidone and 10g of butanediol into the enzymolysis product, carrying out ultrasonic treatment for 20min, then adding 60mL of 80% ethanol water solution with volume concentration, heating, refluxing, extracting, and filtering to obtain a filtrate;
s3, feeding the filtrate into a chitosan adsorption column, wherein the sample loading flow rate is 5BV/h, when the pH of the effluent is 5, leaching the chitosan adsorption column by using an ethanol aqueous solution with the volume concentration of 30% until the effluent is neutral, then eluting by using an ethanol aqueous solution containing sodium hydroxide with the pH of 10, wherein the elution flow rate is 3BV/h, and collecting the effluent;
s4, performing polarity exchange on the effluent liquid by adopting cation exchange resin to obtain a purified liquid, and concentrating and drying the purified liquid to obtain the illicium verum extract;
wherein chitosan in the chitosan adsorption column is fibroin-modified chitosan, and the specific modification method comprises the following steps: dissolving chitosan in acetic acid water solution, adding mixed acid consisting of concentrated nitric acid with the concentration of 90% and concentrated phosphoric acid with the concentration of 85%, and then adding sodium nitrite to react at 20 ℃ to obtain carboxylated chitosan; adding fibroin into the carboxylated chitosan, and reacting at 30 ℃ to obtain modified chitosan; the weight volume ratio of the chitosan to the mixed acid is g: mL is 1:4.5, the volume ratio of concentrated nitric acid to concentrated phosphoric acid in the resultant acid is 2:1, the addition amount of sodium nitrite is 0.7 percent of the total mass of the mixed acid, and the weight ratio of the carboxylated chitosan to the fibroin is 3:0.5.
example 5
A method for extracting natural plant endogenous components comprises the following steps:
s1, drying and crushing fresh pine needles to obtain pine needle powder, adding 30ml of water and pectinase, hemicellulase, protease and cellulase into 10g of pine needle powder, performing enzymolysis for 10min at 30 ℃, and inactivating to obtain an enzymolysis product;
s2, adding 7.5g of polyvinylpyrrolidone and 7.5g of butanediol into the enzymolysis product, carrying out ultrasonic treatment for 20min, then adding 60mL of 80% ethanol water solution with volume concentration, heating, refluxing and extracting, and filtering to obtain a filtrate;
s3, putting the filtrate into a chitosan adsorption column, wherein the sample loading flow rate is 5BV/h, when the pH of the effluent is 5, leaching the chitosan adsorption column by using an ethanol aqueous solution with the volume concentration of 30% until the effluent is neutral, then eluting by using an ethanol aqueous solution containing sodium hydroxide with the pH of 10, wherein the elution flow rate is 3BV/h, and collecting the effluent;
s4, performing polarity exchange on the effluent liquid by adopting cation exchange resin to obtain a purified liquid, and concentrating and drying the purified liquid to obtain a pine needle extract;
wherein chitosan in the chitosan adsorption column is fibroin-modified chitosan, and the specific modification method comprises the following steps: dissolving chitosan in acetic acid water solution, adding mixed acid consisting of concentrated nitric acid with the concentration of 90% and concentrated phosphoric acid with the concentration of 85%, and then adding sodium nitrite to react at 20 ℃ to obtain carboxylated chitosan; adding fibroin into the carboxylated chitosan, and reacting at 30 ℃ to obtain modified chitosan; the weight volume ratio of the chitosan to the mixed acid is g: mL is 1:4.5, the volume ratio of concentrated nitric acid to concentrated phosphoric acid in the resultant acid is 2:1, the addition amount of sodium nitrite is 0.7 percent of the total mass of the mixed acid, and the weight ratio of the carboxylated chitosan to the fibroin is 3:0.5.
comparative example 1
A method for extracting natural plant endogenous components comprises the following steps:
s1, drying and crushing fresh pine needles to obtain pine needle powder, adding 30ml of water and pectinase, hemicellulase, protease and cellulase into 10g of pine needle powder, performing enzymolysis for 10min at 30 ℃, and inactivating to obtain an enzymolysis product;
s2, adding 60mL of 80% ethanol aqueous solution in volume concentration into the enzymolysis product, heating, refluxing, extracting, and filtering to obtain a filtrate;
s3, feeding the filtrate into a chitosan adsorption column, wherein the sample loading flow rate is 5BV/h, when the pH of the effluent is 5, leaching the chitosan adsorption column by using an ethanol aqueous solution with the volume concentration of 30% until the effluent is neutral, then eluting by using an ethanol aqueous solution containing sodium hydroxide with the pH of 10, wherein the elution flow rate is 3BV/h, and collecting the effluent;
s4, performing polarity exchange on the effluent liquid by adopting cation exchange resin to obtain a purified liquid, and concentrating and drying the purified liquid to obtain a pine needle extract;
wherein, the chitosan in the chitosan adsorption column is fibroin modified chitosan, and the specific modification method comprises the following steps: dissolving chitosan in acetic acid water solution, adding mixed acid consisting of concentrated nitric acid with the concentration of 90% and concentrated phosphoric acid with the concentration of 85%, and then adding sodium nitrite to react at 20 ℃ to obtain carboxylated chitosan; adding fibroin into the carboxylated chitosan, and reacting at 30 ℃ to obtain modified chitosan; the weight volume ratio of the chitosan to the mixed acid is g: mL is 1:4.5, the volume ratio of concentrated nitric acid to concentrated phosphoric acid in the resultant acid is 2:1, the addition amount of sodium nitrite is 0.7 percent of the total mass of the mixed acid, and the weight ratio of the carboxylated chitosan to the fibroin is 3:0.5.
comparative example 2
A method for extracting natural plant endogenous components comprises the following steps:
s1, drying and crushing fresh pine needles to obtain pine needle powder, adding 30ml of water and pectinase, hemicellulase, protease and cellulase into 10g of pine needle powder, performing enzymolysis for 10min at 30 ℃, and inactivating to obtain an enzymolysis product;
s2, adding 15g of polyvinylpyrrolidone into the enzymolysis product, carrying out ultrasonic treatment for 20min, then adding 60mL of 80% ethanol aqueous solution by volume, heating, refluxing, extracting, and filtering to obtain a filtrate;
s3, putting the filtrate on a chitosan adsorption column, loading the sample at the flow speed of 5BV/h, when the pH of the effluent is 5, leaching the chitosan adsorption column by using an ethanol aqueous solution with the volume concentration of 30 percent until the effluent is neutral, and then, leaching by using sodium hydroxide-containing solution with the pH of 10
Eluting with ethanol water solution at the flow rate of 3BV/h, and collecting the effluent;
s4, performing polarity exchange on the effluent liquid by adopting cation exchange resin to obtain a purified liquid, and concentrating and drying the purified liquid to obtain a pine needle extract;
wherein, the chitosan in the chitosan adsorption column is fibroin modified chitosan, and the specific modification method comprises the following steps: dissolving chitosan in acetic acid water solution, adding mixed acid consisting of concentrated nitric acid with the concentration of 90% and concentrated phosphoric acid with the concentration of 85%, and then adding sodium nitrite to react at 20 ℃ to obtain carboxylated chitosan; adding fibroin into the carboxylated chitosan, and reacting at 30 ℃ to obtain modified chitosan; the weight volume ratio of the chitosan to the mixed acid is g: mL is 1:4.5, the volume ratio of concentrated nitric acid to concentrated phosphoric acid in the resultant acid is 2:1, the addition amount of sodium nitrite is 0.7 percent of the total mass of the mixed acid, and the weight ratio of the carboxylated chitosan to the fibroin is 3:0.5.
comparative example 3
A method for extracting natural plant endogenous components comprises the following steps:
s1, drying and crushing fresh pine needles to obtain pine needle powder, adding 30ml of water and pectinase, hemicellulase, protease and cellulase into 10g of pine needle powder, performing enzymolysis for 10min at 30 ℃, and inactivating to obtain an enzymolysis product;
s2, adding 15g of butanediol into the enzymolysis product, carrying out ultrasonic treatment for 20min, then adding 60mL of 80% ethanol aqueous solution with volume concentration, heating, refluxing and extracting, and filtering to obtain a filtrate;
s3, the sample loading flow rate is 5BV/h, when the pH of the effluent is 5, the chitosan adsorption column is leached by using an ethanol aqueous solution with the volume concentration of 30% until the effluent is neutral, then the chitosan adsorption column is eluted by using an ethanol aqueous solution containing sodium hydroxide with the pH of 10, the elution flow rate is 3BV/h, and the effluent is collected;
and S4, performing polarity exchange on the effluent liquid by adopting cation exchange resin to obtain a purified liquid, and concentrating and drying the purified liquid to obtain the pine needle extract.
Wherein chitosan in the chitosan adsorption column is fibroin-modified chitosan, and the specific modification method comprises the following steps: dissolving chitosan in acetic acid water solution, adding mixed acid consisting of concentrated nitric acid with the concentration of 90% and concentrated phosphoric acid with the concentration of 85%, and then adding sodium nitrite to react at 20 ℃ to obtain carboxylated chitosan; adding fibroin into the carboxylated chitosan, and reacting at 30 ℃ to obtain modified chitosan; the weight volume ratio of the chitosan to the mixed acid is g: mL is 1:4.5, the volume ratio of concentrated nitric acid to concentrated phosphoric acid in the resultant acid is 2:1, the addition amount of sodium nitrite is 0.7 percent of the total mass of the mixed acid, and the weight ratio of the carboxylated chitosan to the fibroin is 3:0.5.
comparative example 4
A method for extracting natural plant endogenous components comprises the following steps:
s1, drying and crushing fresh pine needles to obtain pine needle powder, adding 30ml of water and pectinase, hemicellulase, protease and cellulase into 10g of pine needle powder, performing enzymolysis for 10min at 30 ℃, and inactivating to obtain an enzymolysis product;
s2, adding 7.5g of polyvinylpyrrolidone and 7.5g of butanediol into the enzymolysis product, carrying out ultrasonic treatment for 20min, then adding 60mL of 80% ethanol water solution with volume concentration, heating, refluxing, extracting, and filtering to obtain a filtrate;
s3, feeding the filtrate into a chitosan adsorption column, wherein the sample loading flow rate is 5BV/h, when the pH of the effluent is 5, leaching the chitosan adsorption column by using an ethanol aqueous solution with the volume concentration of 30% until the effluent is neutral, then eluting by using an ethanol aqueous solution containing sodium hydroxide with the pH of 10, wherein the elution flow rate is 3BV/h, and collecting the effluent;
and S4, performing polarity exchange on the effluent liquid by adopting cation exchange resin to obtain a purified liquid, and concentrating and drying the purified liquid to obtain the pine needle extract.
Examples 2-5 and comparative examples 1-4 the results of measuring the contents of flavonoids and shikimic acid in the endogenous plant components obtained by extraction are shown in the following table:
* The yield is the ratio of the content of a certain effective component in the plant endogenous component of the final product to the content of the effective component in the filtrate; the extraction rate is the gram of a certain effective component extracted from each 100 g of plant powder, wherein the yield in the comparison example 4 is calculated by the content of the effective component in the filtrate, and the yield in other examples and the comparison example is calculated by the effective component in the finally obtained plant endogenous component.
The swelling properties of the chitosan adsorption columns in example 5 and comparative examples 1 to 4 are shown in the following table:
the above description is only a preferred embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications and equivalents of the present invention, which are made by the present specification and directly/indirectly applied to other related technical fields within the spirit of the present invention are included in the scope of the present invention.
Claims (9)
1. A method for extracting natural plant endogenous components is characterized by comprising the following steps:
s1, drying and crushing natural plants containing flavone and shikimic acid to obtain plant powder, adding water and enzyme into the plant powder for enzymolysis, and inactivating to obtain an enzymolysis product;
s2, adding polyvinylpyrrolidone and butanediol into the enzymolysis product, carrying out ultrasonic treatment for 20-40min, then adding an ethanol water solution with the volume concentration of 50-80%, heating, refluxing, extracting, and filtering to obtain a filtrate, wherein the volume ratio of the ethanol water solution to the enzymolysis product is 1-2:1-2;
s3, putting the filtrate into a chitosan adsorption column, when the pH of the effluent is 4-5, leaching the chitosan adsorption column by using an ethanol aqueous solution with the volume concentration of 20-30% until the effluent is neutral, then eluting by using an ethanol aqueous solution containing sodium hydroxide with the pH of 9-10, and collecting the effluent;
s4, purifying, concentrating and drying the effluent to obtain plant endogenous components;
in the step S2, the weight volume ratio g of the total amount of the polyvinylpyrrolidone and the butanediol to the enzymolysis product is as follows: mL is 0.2-0.5:1; the weight ratio of the polyvinylpyrrolidone to the butanediol is 1-2:1-2;
the chitosan is fibroin modified chitosan, and the specific modification method comprises the following steps: dissolving chitosan in acetic acid water solution, adding mixed acid consisting of concentrated nitric acid and concentrated phosphoric acid, then adding sodium nitrite, and reacting at 20-30 ℃ to obtain carboxylated chitosan; adding fibroin into the carboxylated chitosan, and reacting at 30-40 ℃ to obtain modified chitosan;
the weight volume ratio of the chitosan to the mixed acid is g: mL is 1:4-5;
the volume ratio of concentrated nitric acid to concentrated phosphoric acid in the mixed acid is 2:1; the concentration of the concentrated nitric acid is 86% or more, and the concentration of the concentrated phosphoric acid is 83% or more;
the addition amount of the sodium nitrite is 0.5-1% of the total mass of the mixed acid;
the weight ratio of the carboxylated chitosan to the fibroin is 1-3:0.2-0.5.
2. The method for extracting natural plant endogenous components according to claim 1, wherein in step S1, the enzyme is one or a mixture of two or more of cellulase, hemicellulase, protease and pectinase.
3. The method for extracting the natural plant endogenous components according to claim 1, wherein in the step S1, the enzymolysis temperature is 20-40 ℃, and the enzymolysis time is 10-20min.
4. The method for extracting natural plant endogenous ingredients according to claim 1 or 2, wherein in the step S1, the weight ratio of the plant powder to the water is 1:2-5.
5. The method for extracting natural plant endogenous components according to any one of claims 1 to 3, wherein in the step S1, the natural plant containing flavone and shikimic acid is one of ginkgo leaves, pine needles and star anise.
6. A method for extracting natural plant endogenous ingredients according to any one of claims 1 to 3, characterized in that the weight ratio of polyvinylpyrrolidone to butanediol is 1:1.
7. the method for extracting natural plant endogenous components according to any one of claims 1 to 3, wherein in the step S3, the flow rate of sample loading is 5 to 6BV/h, and the flow rate of elution is 3 to 4BV/h.
8. The method for extracting natural plant endogenous components according to any one of claims 1 to 3, wherein in the step S3, the chitosan adsorption column is prepared by the following steps: wetting chitosan with water, and loading into a column to obtain the chitosan adsorption column.
9. The method for extracting natural plant endogenous components according to any one of claims 1 to 3, wherein in step S4, purification is performed by using a cation resin column.
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