CN103550951A - Novel method for extracting or purifying effective components of traditional Chinese medicine - Google Patents

Abstract

A new method for extracting or purifying the active ingredients of traditional Chinese medicine belongs to the field of normal temperature and high pressure processing technology for small molecular biological substances and screening technology for separation through filtration membranes. The hydrostatic pressure of 120MPa~750MPa is kept for 15s, and the solvent penetrates into the solid raw material under the ultra-high pressure, so that the active ingredients of traditional Chinese medicine are dissolved in the solvent, and the dissolution equilibrium is reached in a short time, and the pressure is quickly released at the ultra-high osmotic pressure difference Under this condition, the active ingredient rapidly diffuses into the extraction solvent around the tissue. The extract is separated by centrifugation, and then the membrane is used as the filter medium to filter layer by layer with different cut-offs, the waste liquid is discarded, and the concentrated liquid is finally left to realize the separation and concentration of the extract. This invention can be widely used in the extraction and purification of small molecules in modern bioengineering, the fine processing of modern agriculture, and the modernization/industrial production of traditional Chinese medicine.

Description

A New Method of Extracting or Purifying Active Components of Traditional Chinese Medicine

technical field

The invention belongs to the technical field of normal-temperature and high-pressure processing technology for small molecular biological substances and separation through filter membranes.

Background technique

The biological high-pressure processing technology in the prior art is obtained by applying a hydrostatic pressure of 120 to 750 MPa to the biological material at room temperature or at a lower temperature (usually lower than 90°C), so that the biological material undergoes physical or chemical changes. A technology to a new product.

Ultra-high pressure daily work can cause changes in macromolecular substances, such as protein denaturation, enzyme inactivation, cell membrane rupture, bacterial component leakage leading to cessation of life activities, microbial cell destruction and death, etc., but for small molecular substances (such as vitamins, Pigments, alkaloids, soaps, flavonoids, etc.) have no effect. According to this principle, the extraction of active ingredients of traditional Chinese medicine at room temperature and ultra-high pressure is proposed.

Membrane separation technology is a screening process related to the pore size of the membrane. The pressure difference on both sides of the membrane is used as the driving force, and the membrane is used as the filter medium. The many tiny micropores in the membrane only allow water and small molecular substances to pass through and become the permeate, while the substances in the original solution whose volume is larger than the micropore diameter of the membrane surface are trapped on the liquid inlet side of the membrane and become concentrated solution, thus realizing the recovery of the original solution. Separate and concentrate. At present, the types of membranes that have been developed in the world include: microfiltration membranes, ultrafiltration membranes, reverse osmosis membranes, etc., and the scope of application has also developed from a single salt water chemical to pure water preparation, wastewater treatment, biology and medicine. Product purification, organic and inorganic separation, food processing, environmental protection, precious metal extraction and gas separation and other fields. In recent years, the application of membrane separation technology to the separation and purification of traditional Chinese medicine extracts has great development potential and application value in terms of improving product yield, purity, reducing energy consumption, processing time and process improvement.

Contents of the invention

In order to solve the existing deficiencies in the prior art regarding the extraction and purification of active ingredients of traditional Chinese medicines, the invention provides an effective extraction method.

A method for extracting and purifying active ingredients of traditional Chinese medicines is characterized in that:

(1) Extract the active ingredients of traditional Chinese medicine from Chinese medicinal plants: first crush the raw plant, the crushed particles are 75 mesh, then add the organic solvent aqueous solution I whose weight is 10 to 20 times that of the raw material powder of traditional Chinese medicine and mix well to obtain the traditional Chinese medicine powder and organic The mixed solution II after the solvent aqueous solution I is fully mixed; then put the mixed solution II into the pressure processor, raise the pressure to 120MPa~750MPa within 5 minutes, keep the pressure for 15 minutes and release the pressure quickly, and release it to normal pressure within 5s , take out the mixed solution II; the organic solvent in the described organic solvent aqueous solution I includes methanol, ethanol, isopropanol, n-butanol, acetone, methyl acetate, ethyl acetate, and the pressure range is 120MPa to 750MPa ;

(2) Coarsely separate the active ingredients of traditional Chinese medicine: Centrifuge the mixed solution II to remove the Chinese medicine dregs in the mixed solution II after centrifugation, and then filter the centrifuged liquid through a medium with a filtration accuracy of 1.0 μm to obtain Remove impurities with a particle size greater than 1.0 μm and fine Chinese medicine dregs in the mixed solution II to obtain a clarified active ingredient extract III of traditional Chinese medicine; (3) Purification of active ingredients of traditional Chinese medicine: filter the extract III through a medium with a filtration accuracy of 0.1 μm Filtration to remove macromolecules with a particle size greater than 0.1 μm in the extract III, and then filter the filtrate through ultrafiltration to filter out the organic macromolecules with a molecular weight greater than 5000-10000 Daltons in the extract III that have no drug effect Molecular substances, to obtain the high-purity active ingredient extract IV of traditional Chinese medicine; the ultrafiltration filtration, the adopted ultrafiltration membrane is selected with a cut molecular weight of 5000-10000 Daltons depending on the treatment of traditional Chinese medicines; the ultrafiltration filtration can be Select ultrafiltration membranes with different cut molecular weights according to the molecular weight of the active ingredients of the extracted traditional Chinese medicine;

(4) Further purification and concentration of active ingredients of traditional Chinese medicine: pass the extract IV through a nanofiltration membrane to remove the organic solvent in the organic solvent aqueous solution I in the extract IV, so as to achieve the purpose of purification and concentration; The nanofiltration membrane used is selected to cut the nanofiltration membrane with a molecular weight of 200-500 Daltons according to the molecular weight of the organic solvent in the organic solvent aqueous solution I.

Beneficial effects: the invention can be widely used in modern bioengineering, modern agricultural engineering, modernized and industrialized production of traditional Chinese medicine. Extraction and purification of small molecules in modern bioengineering; fine processing of modern agriculture, such as extraction, purification and industrial production of small molecules such as soybeans and corn; extraction and purification of most Chinese herbal medicines. Compared with traditional techniques, the application of the present invention has two advantages: first, compared with traditional techniques such as decoction, reflux, and Soxhlet extraction, the high-pressure processing technique can greatly shorten the extraction time and reduce energy consumption, and the high-pressure day labor is in the It is carried out at room temperature, which solves the problems of structural change, loss and physiological activity reduction of active ingredients caused by high temperature, effectively maintains its active ingredients, and greatly improves the medical and health care effects; It is carried out in a closed environment, and the solvent will not volatilize into the surrounding environment, which meets the requirements of "green" environmental protection. Therefore, the application of high-pressure processing technology to extract the active ingredients of traditional Chinese medicine has high practical value and is a new technology with great development potential and competitiveness. Secondly, it has shown great development potential and application value in terms of improving product yield, purity, reducing energy consumption, processing time and process improvement.

Detailed ways

The present invention applies a hydrostatic pressure of 120MPa to 750MPa rapidly to the mixed liquid of traditional Chinese medicine raw material powder and extract under the condition of normal temperature or lower temperature (usually lower than 90°C), and keeps the pressure for 15 minutes. Rapidly penetrate into the interior of the solid raw material, the active ingredient dissolves in the solvent, and reaches the dissolution equilibrium in a short time, and then quickly releases the pressure. Under the ultra-high osmotic pressure difference, the active ingredient quickly diffuses into the extraction solvent around the tissue, thereby achieving Extraction of active ingredients of traditional Chinese medicine. Then adopt the method of membrane separation to carry out step-by-step separation to remove impurities and macromolecular substances without medicinal value layer by layer, so as to complete the purpose of removing ineffective components in the solution. Finally, the solvent in the extract is removed, so that the active ingredients of the traditional Chinese medicine in the extract are concentrated.

First carry out the extraction of the active ingredients of the traditional Chinese medicine: the traditional Chinese medicine is first pulverized, and the pulverized particles are about 60 mesh (depending on the variety and characteristics of the Chinese medicine). Mix the pulverized traditional Chinese medicine raw materials into the extract (such as ethanol, methanol, isopropanol, n-butanol, acetone, methyl acetate, ethyl acetate, etc., depending on the extract, choose the extract) and mix well Finally, put it into the ultra-high pressure adding device, add 120MPa to 750MPa high pressure (depending on the extraction raw material, select the pressure), keep the pressure for 15 minutes, and take out the mixed extract. At this time, under high pressure, the active ingredients of the traditional Chinese medicine have been dissolved into the extract.

Subsequently, the active ingredients of the traditional Chinese medicine are purified, and the mixed extract is centrifuged to remove the dregs of the traditional Chinese medicine. Then, the extract after centrifugation is subjected to coarse filtration, and the filter membrane has a pore size of 1.0 μm, which can remove impurities and fine Chinese medicine residues in the mixed extract. After coarse filtration the mixture became clear. Then the clarified mixture is subjected to microfiltration to remove macromolecular impurities in the extract. Then select ultrafiltration membranes with different cut-offs according to the raw materials of Chinese medicinal materials and extraction requirements, and concentrate the extract by ultrafiltration. During the ultrafiltration process, the concentrated solution can be repeatedly filtered to increase the concentration factor. The filtrate can be properly disposed of according to the situation, can be thrown away directly, or can be filtered again to increase the yield. If it is necessary to increase the concentration multiple, according to the different characteristics of the extracted liquid, use nanofiltration or reverse osmosis to further concentrate.

Embodiment one

Extracting Total Flavonoids from Epimedium

Epimedium is a natural medicinal plant with important pharmacological effects. The flavonoids contained in Epimedium, especially the main active ingredient icariin, have a wide range of physiological activities and can be used for blood lipid regulation, antihypertensive, anti Inflammation, anti-osteoporosis, anti-aging, anti-tumor, etc.

(1) After crushing the dry raw material of Epimedium Fructus into 60 mesh powder, mix it with an aqueous solution of 60% ethanol of 15 times its weight. After fully stirring and mixing, put the epimedium powder and ethanol aqueous solution into the ultra-high pressure container, raise the pressure to 650MPa within 5 minutes, keep the pressure for 10 minutes, quickly release the pressure, and reduce the pressure to normal pressure within 5 seconds.

(2) Centrifuge the epimedium powder and ethanol aqueous solution after ultra-high pressure treatment, and discard the dregs after separation. Then the separated extract is subjected to coarse filtration, the pore size of the fine filter membrane is 1.0 μm, and the working temperature is room temperature, to remove impurities and residual medicinal residues in the extract to obtain a clarified Epimedium extract.

Pass the epimedium extract through a honeycomb microfilter with a pore size of 0.1 μm to mainly remove macromolecular impurities in the extract. The filtrate is then subjected to ultrafiltration. Ultrafiltration adopts an ultrafiltration membrane with a molecular weight cut-off of 10,000 Daltons. The working pressure of the ultrafiltration is 0.1MPa, and the working temperature is normal temperature. The ultrafiltered extract has removed the ineffective components in the extract that are greater than the molecular weight of the total flavonoids of Epimedium. At this time, the extract is a dilute solution of total flavonoids of Epimedium radix with high purity.

The dilute solution of the total flavonoids of Epimedium is concentrated by nanofiltration, so as to remove the solvent in the total flavonoids of Epimedium. When filtering, the working temperature is normal temperature, the working pressure is 7MPa, and the molecular weight cut-off of the nanofiltration membrane is 500 Daltons. After nanofiltration, the concentrated epimedium total flavonoids solution is obtained.

Conclusion: After ultra-high pressure extraction and membrane purification and concentration, the yield of total flavonoids of Epimedium radix effective ingredients is 78.3%, and the purity of total flavonoids is 44.2%.

Embodiment two

Glycyrrhizic acid extracted from licorice

Licorice is a natural medicinal plant with important pharmacological effects. Glycyrrhizic acid contained in licorice, also known as glycyrrhizin saponin, is the main active ingredient in licorice. It has anti-inflammatory, anti-viral, liver-protecting and detoxifying properties. It has an inhibitory effect, and the inhibition rate of AIDS is as high as 80%, and it has a strong effect of increasing the immune function of the human body.

(1) After the dry raw material of licorice is crushed into 60 mesh powder, mix it with an aqueous solution of 60% ethanol in an amount 15 times its weight. After fully stirring and mixing, put the licorice powder and ethanol aqueous solution into the ultra-high pressure container, raise the pressure to 450MPa within 8 minutes, keep the pressure for 15 minutes, quickly release the pressure, and reduce the pressure to normal pressure within 5 seconds.

(2) Centrifuge the licorice powder after ultra-high pressure treatment and ethanol aqueous solution, and discard the medicinal residue after separation. Then the separated extract is subjected to coarse filtration, the fine filter membrane has a pore size of 1 μm, and the working temperature is room temperature to remove impurities and residual medicinal residues in the extract to obtain a clarified licorice extract.

(3) The licorice extract is passed through a honeycomb microfilter with a pore size of 0.1 μm to mainly remove macromolecular impurities in the extract. The filtrate is then subjected to ultrafiltration. Ultrafiltration adopts an ultrafiltration membrane with a molecular weight cut-off of 10,000 Daltons. The working pressure of the ultrafiltration is 0.1MPa, and the working temperature is normal temperature. When the extract is a dilute solution of glycyrrhizic acid with high purity.

(4) Concentrate the dilute solution of glycyrrhizic acid by nanofiltration to remove the solvent in the glycyrrhizic acid solution. When filtering, the working temperature is normal temperature, the working pressure is 6MPa, and the molecular weight cut-off of the nanofiltration membrane is 500 Daltons. After nanofiltration, a concentrated glycyrrhizic acid solution was obtained.

Conclusion: After ultra-high pressure extraction and membrane purification and concentration, the active ingredient of glycyrrhizic acid was obtained at a rate of 67.6%, and the purity of glycyrrhizic acid was 70.2%.

Claims (1)

1. A new method for extracting or purifying active ingredients of traditional Chinese medicine, characterized in that: (1) Extract the active ingredients of traditional Chinese medicine from Chinese medicinal plants: first crush the raw plant, the crushed particles are 75 mesh, then add the organic solvent aqueous solution I whose weight is 10 to 20 times that of the raw material powder of traditional Chinese medicine and mix well to obtain the traditional Chinese medicine powder and organic The mixed solution II after the solvent aqueous solution I is fully mixed; then put the mixed solution II into the pressure processor, raise the pressure to 120MPa~750MPa within 5 minutes, keep the pressure for 15 minutes and release the pressure quickly, and release it to normal pressure within 5s , take out mixed solution II; the organic solvent in the described organic solvent aqueous solution I comprises methanol, ethanol, Virahol, n-butanol, propane, methyl acetate, ethyl acetate, and the pressure range is 120MPa? 750MPa; (2) Coarsely separate the active ingredients of traditional Chinese medicine: Centrifuge the mixed solution II to remove the Chinese medicine dregs in the mixed solution II after centrifugation, and then filter the centrifuged liquid through a medium with a filtration accuracy of 1.0 μm to obtain Remove impurities with a particle size greater than 1.0 μm and fine Chinese medicine dregs in the mixed solution II to obtain a clarified active ingredient extract III of traditional Chinese medicine; (3) Purification of active ingredients of traditional Chinese medicine: filter the extract III through a medium with a filtration accuracy of 0.1 μm Filtration to remove macromolecules with a particle size greater than 0.1 μm in the extract III, and then filter the filtrate through ultrafiltration to filter out the organic macromolecules with a molecular weight greater than 5000-10000 Daltons in the extract III that have no drug effect Molecular substances, to obtain the high-purity active ingredient extract IV of traditional Chinese medicine; the ultrafiltration filtration, the adopted ultrafiltration membrane is selected with a cut molecular weight of 5000-10000 Daltons depending on the treatment of traditional Chinese medicines; the ultrafiltration filtration can be Select ultrafiltration membranes with different cut molecular weights according to the molecular weight of the active ingredients of the extracted traditional Chinese medicine; (4) Further purification and concentration of the active ingredients of traditional Chinese medicine: pass the extract IV through a nanofiltration membrane to remove the organic solvent in the organic solvent aqueous solution I in the extract IV, so as to achieve the purpose of purification and concentration; said uranium filtration, the The adopted uranium filter membrane selects a nanofiltration membrane with a molecular weight of 200-500 Daltons according to the molecular weight of the organic solvent in the organic solvent aqueous solution I.