CN114917265B - Extraction method of ginsenoside - Google Patents

Extraction method of ginsenoside Download PDF

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CN114917265B
CN114917265B CN202210803694.7A CN202210803694A CN114917265B CN 114917265 B CN114917265 B CN 114917265B CN 202210803694 A CN202210803694 A CN 202210803694A CN 114917265 B CN114917265 B CN 114917265B
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ginsenoside
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CN114917265A (en
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何洁
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Wuyang County Center For Disease Control And Prevention
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    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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    • A—HUMAN NECESSITIES
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
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    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract

The invention relates to the field of medicines and discloses a method for extracting ginsenoside. The extraction method comprises the following steps: (1) Mixing ginseng raw material with an acidic solution for acidizing treatment, and then mixing with a first extraction solvent for reflux extraction to obtain a first extraction solution; (2) Mixing the first extract with a second extraction solvent and an auxiliary agent, performing reflux extraction II to separate ginsenoside, and performing solid-liquid separation I to obtain a second extract; (3) Purifying the second extract, and drying to obtain ginsenoside extract; wherein the auxiliary agent in the step (2) is magnetic nano particles and/or activated clay. The method can effectively improve yield and content of ginsenoside.

Description

Extraction method of ginsenoside
Technical Field
The invention relates to the field of medicines, in particular to a method for extracting ginsenoside.
Background
Ginseng is a dry root of ginseng of the Araliaceae plant, is a third-stage wiggle plant, belongs to a rare tonic, is mainly produced in east Asian, is a traditional rare traditional Chinese medicine in China, and has a medicinal foundation for thousands of years. In Ben Cao gang mu, ginseng radix can treat spontaneous perspiration, dizziness, headache, regurgitation, diarrhea, frequent urination, dribbling, tiredness, internal injury, apoplexy, heatstroke, arthralgia, hematemesis, hemoptysis, metrorrhagia, and antenatal and postnatal diseases. Ginseng is commonly used in modern traditional Chinese medicine for tonifying qi, rescuing collapse, nourishing blood, restoring pulse, nourishing heart, soothing nerves, promoting the production of body fluid, quenching thirst, tonifying lung, relieving asthma, strengthening spleen, relieving diarrhea, expelling toxin, curing sore and the like. The ginseng contains ginsenoside, polysaccharide, organic acid, volatile oil, protein, amino acid, vitamin, alkaloid, lipidosome, sterol, lignin and other components. Ginsenoside is the most main active ingredient in ginseng, belongs to a solid alcohol type triterpenoid saponin compound, and has obvious effects of resisting cancer, resisting inflammation, regulating immune system, resisting hypertension, resisting diabetes, strengthening spleen, tonifying qi and the like.
CN111450132a discloses a method for extracting ginsenoside by a bionic method, which specifically comprises the following steps: selecting ginseng planted for more than 4 years, washing with water until no sludge is on the surface, drying the ginseng to constant weight at 45 ℃ by using a blast drying oven, crushing the dried ginseng by using a crusher, and sieving the crushed ginseng with a 60-mesh sample separating sieve; 1.64mL of dilute hydrochloric acid solution of 2.34mol/L is added into 80mL of deionized water, 1g of pepsin powder is weighed and dissolved in the dilute hydrochloric acid solution, deionized water is used for fixing the volume to 100mL, and 10mg/mL of bionic gastric juice is obtained, wherein the specific activity of the pepsin is not lower than 1:3000; taking 1g of ginseng powder and 15mL of bionic gastric juice, heating and refluxing at 37 ℃ and normal pressure, extracting for 60 minutes, centrifuging at 8000rpm for 8 minutes, and separating an extracting solution and ginseng powder; the volume ratio of the extraction solvent to the extraction solution is 1:1, extracting for 2 times, mixing the extracts, and volatilizing to obtain ginsenoside extract.
CN110526947a discloses a preparation method of ginsenoside extract for beverage, which comprises the following steps: firstly, extracting ginsenoside in raw material ginseng in liquid state, removing grease in ginsenoside extraction filtrate by utilizing diatomite, then filtering to remove insoluble tiny impurities in the ginsenoside extraction filtrate by a three-stage filtering system, finally, performing spray drying, combining the difference of protein molecular weight and ginsenoside molecular weight with ultrafiltration membrane separation, and separating unstable macromolecular protein, thereby obtaining the ginsenoside extract with high solution stability.
However, the yield and content of ginsenoside obtained by the method for obtaining ginsenoside in the prior art still need to be further improved.
Disclosure of Invention
The invention aims to solve the problem that the yield and the content of ginsenoside extracted from ginseng in the prior art are to be further improved, and provides a method for extracting ginsenoside, which can effectively improve the yield and the content of ginsenoside.
In order to achieve the above object, the present invention provides a method for extracting ginsenoside, comprising the steps of:
(1) Mixing ginseng raw material with an acidic solution for acidizing treatment, and then mixing with a first extraction solvent for reflux extraction to obtain a first extraction solution;
(2) Mixing the first extract with a second extraction solvent and an auxiliary agent, performing reflux extraction II to separate ginsenoside, and performing solid-liquid separation I to obtain a second extract;
(3) Purifying the second extract, and drying to obtain ginsenoside extract;
wherein the auxiliary agent in the step (2) is magnetic nano particles and/or activated clay.
Preferably, in the step (1), the ginseng raw material is prepared by drying and pulverizing ginseng.
Preferably, the conditions of the acidification treatment comprise: the pH is 2-2.5, the temperature is 30-45deg.C, and the time is 20-30min.
Preferably, the acidic solution is an organic acid solution, more preferably an acetic acid solution.
Preferably, the first extraction solution in step (1) is selected from at least one of methanol, ethanol and water, more preferably ethanol and/or methanol.
Preferably, the weight ratio of the ginseng raw material to the first extraction solvent is 1:15-20.
Preferably, the conditions for reflux extraction I in step (1) include: the temperature is 85-110 ℃ and the time is 3-6h.
Preferably, the second extraction solvent in step (2) is water.
Preferably, before mixing the first extract with the second extraction solvent and the auxiliary agent, the method further comprises concentrating the first extract under reduced pressure to remove the first extraction solvent.
Preferably, in the step (2), the weight ratio of the first extracting solution, the second extracting solvent and the auxiliary agent is 1:5-10:0.5-1.
Preferably, the auxiliary agent is magnetic nano particles and activated clay.
Preferably, the weight ratio of the magnetic nanoparticles to the activated clay is 3-4:1.
Preferably, the magnetic nanoparticles in step (2) are ferroferric oxide nanoparticles.
Preferably, the preparation method of the auxiliary agent comprises the following steps: in the presence of the activated clay, carrying out a contact reaction between an iron salt solution containing iron ions and ferrous ions and an alkaline solution, and then carrying out solid-liquid separation II and washing.
Preferably, the conditions of the contact reaction include: under the anaerobic condition, the pH value is 10-12, the temperature is 50-80 ℃ and the time is 1-6h.
Preferably, in step (2), the conditions for reflux extraction II include: the temperature is 85-100deg.C, and the time is 1-2h.
Preferably, the solid-liquid separation I adopts ultrafiltration membrane filtration, and the filtration precision of the ultrafiltration membrane is 0.01-0.1 mu m.
Preferably, the purification treatment in step (3) comprises: and (3) performing silica gel column chromatography on the second extract, and collecting the eluent.
Preferably, the eluent of the silica gel column chromatography is methanol-water solution.
Through the technical scheme, the invention has the beneficial effects that:
according to the extraction method provided by the invention, the acidification treatment of the ginseng raw material and the auxiliary agent during extraction are combined to form a mutual synergistic effect, the leaching efficiency of the ginsenoside in the ginseng raw material is effectively improved through the acidification treatment, and further, the magnetic nano particles and/or activated clay are used as the auxiliary agent during the extraction of the second extraction solvent, so that the liquid turbulence in the reflux extraction II process can be enhanced, the leaching of the ginsenoside into the second extraction solvent is accelerated, and the extraction yield and efficiency of the ginsenoside are effectively improved.
The extraction method provided by the invention combines ultrafiltration membrane filtration adopted by the solid-liquid separation I with silica gel column chromatography during purification treatment, and can further improve the yield and content of ginsenoside.
Additional features and advantages of the invention will be set forth in the detailed description which follows.
Detailed Description
The endpoints and any values of the ranges disclosed herein are not limited to the precise range or value, and are understood to encompass values approaching those ranges or values. For numerical ranges, one or more new numerical ranges may be found between the endpoints of each range, between the endpoint of each range and the individual point value, and between the individual point value, in combination with each other, and are to be considered as specifically disclosed herein.
The invention provides a method for extracting ginsenoside, which comprises the following steps:
(1) Mixing ginseng raw material with an acidic solution for acidizing treatment, and then mixing with a first extraction solvent for reflux extraction to obtain a first extraction solution;
(2) Mixing the first extract with a second extraction solvent and an auxiliary agent, performing reflux extraction II to separate ginsenoside, and performing solid-liquid separation I to obtain a second extract;
(3) Purifying the second extract, and drying to obtain ginsenoside extract;
wherein the auxiliary agent in the step (2) is magnetic nano particles and/or activated clay.
The extraction method provided by the invention combines the acidification treatment of the ginseng raw material with the auxiliary agent during extraction to form a mutual synergistic effect; the leaching efficiency of ginsenoside in the ginseng raw material is effectively improved through acidification treatment, and further, when the second extraction solvent is used for extraction, magnetic nano particles and/or activated clay are used as auxiliary agents, so that the liquid turbulence in the reflux extraction II process can be enhanced, the leaching of the ginsenoside into the second extraction solvent is accelerated, and the extraction yield and efficiency of the ginsenoside are effectively improved.
According to the invention, the ginseng raw materials can be selected from the rhizome, the ginseng rootlet, the ginseng bud, the ginseng leaf, the ginseng flower and the ginseng fruit, and are all commercially available.
According to the present invention, preferably, in the step (1), the ginseng raw material is prepared by drying and pulverizing ginseng.
According to the present invention, the acidification treatment is to soak and acidify the ginseng raw material in an acidic solution, preferably, in order to further increase the leaching rate of the ginsenoside in the first extraction solvent, the acidification treatment conditions include: the pH is 2-2.5, and can be specifically 2, 2.1, 2.2, 2.3, 2.4, 2.5 or any value between the two values; the temperature is 30-45deg.C, specifically 30deg.C, 33deg.C, 36deg.C, 39deg.C, 42deg.C, 45deg.C, or any value between the above two values; the time is 20-30min, specifically 20min, 22min, 24min, 26min, 28min, 30min, or any value between the above two values.
According to the present invention, the acidic solution may be any acidic solution in the art, for example, hydrochloric acid solution, sulfuric acid solution, nitric acid solution, acetic acid solution, citric acid solution. In order to further optimize the effect of the acidification treatment on the ginseng raw material and improve the extraction efficiency of the ginsenoside, the acidic solution is preferably an organic acid solution, and more preferably an acetic acid solution. During the research process, the inventor surprisingly finds that the effect of the acidification treatment of the ginseng raw material by adopting the organic acid solution is far better than the acidification effect of the inorganic acid solution, thereby being beneficial to improving the leaching rate of the ginsenoside in the first extracting solution.
In the present invention, the weight ratio of ginseng raw material to acidic solution can be adjusted within a wide range, and preferably, the weight ratio of ginseng raw material to acidic solution is 1:2-4.
In the invention, the acidized solution is directly mixed with the first extraction solvent for reflux extraction of I, so that the process of reflux extraction of I is further under an acidic condition, and the extraction effect of the first extraction solvent on the ginsenoside is better.
According to the present invention, the first extraction solution may be an extraction solvent conventional in the art, preferably, the first extraction solution in step (1) is selected from at least one of methanol, ethanol and water, more preferably ethanol and/or methanol.
According to the present invention, in order to further enhance the extraction and leaching effect of the first extraction solvent on ginsenoside and further enhance the yield of ginsenoside, it is preferable that the weight ratio of the ginseng raw material to the first extraction solvent is 1:15-20.
According to the present invention, the conditions for the reflux extraction I may be conventional in the art, for example, the conditions for the reflux extraction I in step (1) include: the temperature is 85-110deg.C, and can be 85deg.C, 90deg.C, 95deg.C, 100deg.C, 105deg.C, 110deg.C, or any value between the above two values; the time is 3-6h, and can be specifically 3h, 4h, 5h, 6h, or any value between the two values.
According to the present invention, in the step (2), the filtrate obtained by solid-liquid separation of the solution obtained by reflux extraction I may be directly mixed with the second extraction solvent and the auxiliary agent, or the filtrate may be concentrated to remove the first organic solvent to obtain a concentrate as the first extraction liquid, and then mixed with the second extraction solvent and the auxiliary agent. The latter is preferable in the present invention, that is, the solution obtained by the reflux extraction I is concentrated under reduced pressure to remove 80 to 90% by volume of the first extraction solvent, and then mixed with the second extraction solvent and the auxiliary agent. Wherein, the conditions of the reduced pressure concentration may be conventional concentration conditions in the art.
According to the present invention, the second extraction solvent may be a solvent capable of separating ginsenoside, which is conventional in the art, and preferably, the second extraction solvent in the step (2) is water. The inventor of the invention finds that in the research process, the first extraction solvent is methanol and/or ethanol, the second extraction solvent is water, so that the extraction effect of the organic solvent and the water on the ginsenoside can be combined, and the content of the ginsenoside can be effectively improved.
According to the present invention, preferably, the weight ratio of the first extract, the second extract and the auxiliary agent in the step (2) is 1:5-10:0.5-1. Here, the first extract is preferably a filtrate obtained by solid-liquid separation of the solution obtained by reflux extraction I, and the filtrate is concentrated to remove the concentrate after the first organic solvent.
According to the present invention, although the efficiency of separating and extracting ginsenoside from ginseng raw material can be improved only in the case where magnetic nanoparticles or activated clay are used as an auxiliary agent, it is preferable that the yield and content of ginsenoside can be further improved when the auxiliary agent is magnetic nanoparticles or activated clay.
According to the present invention, the mixing ratio of the magnetic nanoparticles to the activated clay may be adjusted within a wide range, preferably, the amount of the magnetic nanoparticles is larger than the amount of the activated clay, and more preferably, the weight ratio of the magnetic nanoparticles to the activated clay is 3-4:1.
According to the present invention, the magnetic nanoparticle is at least one selected from the group consisting of an oxide nanoparticle of iron, an oxide nanoparticle of cobalt, and an oxide nanoparticle of nickel. Preferably, the magnetic nanoparticles in step (2) are ferroferric oxide nanoparticles.
The active clay is prepared with clay as main material, and through inorganic acidification, water rinsing and drying.
According to the present invention, the ferroferric oxide nanoparticles are commercially available, and can also be prepared by methods disclosed in the prior art. Illustratively, the method of preparing the ferroferric oxide nanoparticle comprises: and performing coprecipitation reaction on the ferric salt solution containing iron ions and ferrous ions and the alkaline solution. The ferroferric oxide nano particles prepared by the coprecipitation method are more uniform in granularity, better in extraction effect on ginsenoside, and capable of effectively removing particle impurities in the first extracting solution, so that the content of the ginsenoside is further improved.
In the present invention, the iron ion and the ferrous ion may be provided by conventional soluble iron salts, soluble ferrous salts, respectively, for example, iron ions are provided by ferric sulfate, ferric nitrate, ferric chloride, and ferrous ions are provided by ferrous sulfate, ferrous nitrate, ferrous chloride, all of which are commercially available. Preferably, the molar ratio of iron ions to ferrous ions in the ferric salt solution is 0.8-1.2:1.
according to the present invention, the solid-liquid separation II may employ a separation method conventional in the art, for example, centrifugation, filtration, suction filtration, etc.
According to the present invention, preferably, the conditions of the contact reaction include: under anaerobic conditions, the pH is 10-12, and can be specifically 10, 10.5, 11, 11.5 and 12, or any value between the two values; the temperature is 50-80deg.C, specifically 50deg.C, 55deg.C, 60deg.C, 65deg.C, 70deg.C, 75deg.C, 80deg.C, or any value between the above two values; the time is 1-6h, and can be specifically 1h, 2h, 3h, 4h, 5h, 6h, or any value between the two values.
According to the invention, the auxiliary agent can be prepared by mixing the magnetic nano particles and the activated clay with the first extracting solution and the second extracting solvent independently, however, the inventor of the invention also found that the extraction effect on ginsenoside can be further optimized and the yield and content of the final ginsenoside can be improved when the auxiliary agent particles are formed by the magnetic nano particles and the activated clay according to the following method and then mixed with the first extracting solution and the second extracting solvent. Specifically, the preparation method of the auxiliary agent comprises the following steps: in the presence of activated clay, carrying out a contact reaction between ferric salt solution containing iron ions and ferrous ions and alkaline solution, and then carrying out solid-liquid separation II and washing, so that at least part of ferroferric oxide nano particles are wrapped on the surface of the activated clay to form a particle structure with micropores, and the ferroferric oxide nano particles are used as an auxiliary agent.
According to the present invention, in the step (2), the conditions for the reflux extraction II include: the temperature is 85-100deg.C, and can be 85deg.C, 90deg.C, 95deg.C, 100deg.C, or any value between the above two values; the time is 1-2h, and can be specifically 1h, 1.2h, 1.4h, 1.6h, 1.8h, 2h, or any value between the two values.
According to the present invention, preferably, the solid-liquid separation I in the step (2) is performed by ultrafiltration membrane having a filtration accuracy of 0.01 to 0.1. Mu.m. The ultrafiltration membrane is a high molecular semipermeable membrane which is used for separating high molecular colloid or suspended particles with a certain size from solution in the ultrafiltration process, and the ultrafiltration membrane filtration generally adopts a filter pressing mode so as to effectively separate the second extract containing the ginsenoside from the auxiliary agent.
According to the present invention, preferably, the purification treatment in step (3) includes: and (3) performing silica gel column chromatography on the second extract, and collecting the eluent.
According to the present invention, preferably, the eluent of the silica gel column chromatography is a methanol-water solution. The concentration of methanol in the eluent can be selected within a wide range, preferably the concentration of methanol is 50 to 90% by volume, more preferably 70 to 80% by volume.
According to a particularly preferred embodiment of the present invention, the extraction method of ginsenoside comprises the following steps:
(1) Drying and crushing ginseng to obtain ginseng raw material, mixing the ginseng raw material and an acidic solution in a weight ratio of 1:2-4, performing acidification treatment for 20-30min at a pH of 2-2.5 and a temperature of 30-45 ℃ to obtain an acidified solution, mixing the acidified solution with a first extraction solvent (the weight ratio of the ginseng raw material to the first extraction solvent is 1:15-20), performing reflux extraction for 3-6h at a temperature of 85-110 ℃, concentrating filtrate obtained by solid-liquid separation of the solution obtained by the reflux extraction to remove a concentrate obtained by the first organic solvent to obtain a first extraction solution;
(2) Under the condition that activated clay exists and no oxygen exists, ferric salt solution containing iron ions and ferrous ions and alkaline solution are subjected to contact reaction for 1-6 hours at the pH value of 10-12 and the temperature of 50-80 ℃, solid-liquid separation II is carried out, an auxiliary agent (the weight ratio of magnetic nano particles to activated clay is 3-4:1) is obtained by washing, the first extract, a second extraction solvent and the auxiliary agent are mixed according to the weight ratio of 1:5-10:0.5-1, reflux extraction is carried out for 1-2 hours at the temperature of 85-100 ℃ to separate ginsenoside, and the second extract is obtained by ultrafiltration membrane filtration (the filtration precision is 0.01-0.1 mu m);
(3) Subjecting the second extract to silica gel column chromatography, eluting with 50-90 vol% methanol-water solution, collecting eluate, and drying the eluate to obtain ginsenoside extract.
The present invention will be described in detail by examples.
In the following examples and comparative examples, ginseng raw material was obtained by drying and pulverizing purchased ginseng rootstock obtained from Changbai mountain area of Jilin province; the ferroferric oxide nanoparticles were purchased from beijing co island gold technologies limited; unless otherwise indicated, the remaining reagents and starting materials were commercially available; the test methods used in the following examples and comparative examples, unless otherwise specified, are conventional test methods well known to those skilled in the art.
In the following examples and comparative examples, the content of ginsenoside in the ginsenoside extract was determined by high performance liquid chromatography-mass spectrometry, specifically, the ginsenoside extract was redissolved with chromatographic methanol, and analyzed by high performance liquid chromatography-mass spectrometry after passing through a PTFE filter membrane with a pore size of 0.22 μm;
yield of ginsenoside = weight mg of ginsenoside extract/weight g of ginseng raw material.
Example 1
(1) Mixing 100g of ginseng raw material with 400g of acetic acid solution (acetic acid concentration is 2mol/L, pH and is 2.23), performing acidification treatment at 35 ℃ for 25min to obtain an acidified solution, mixing the acidified solution with 1.8kg of ethanol, performing reflux extraction at 100 ℃ for 5h, filtering the solution obtained by reflux extraction to obtain a filtrate, and performing rotary evaporation concentration on the filtrate at 50 ℃ to remove about 90 vol% of ethanol to obtain 456g of concentrate as a first extract;
(2) Mixing 100g of activated clay, a ferric salt solution containing 0.75mol of ferric sulfate and 0.75mol of ferrous sulfate with a proper amount of sodium hydroxide solution under the anaerobic condition, carrying out contact reaction for 4 hours under the conditions of pH 11 and temperature 65 ℃ to obtain a reaction solution, washing a solid substance obtained after filtering the reaction solution with water, and drying to obtain an auxiliary agent (the weight ratio of magnetic nano particles to activated clay is about 3.5:1); mixing the first extract with 3.64kg of water and 364g of auxiliary agent, reflux-extracting at 90deg.C for 1.5 hr to separate ginsenoside, and filtering with ultrafiltration membrane (filtering accuracy is 0.05 μm) to obtain second extract;
(3) Subjecting the second extract to silica gel column chromatography, eluting with 75% methanol-water solution, collecting eluate, concentrating, and spray drying to obtain ginsenoside extract.
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Example 2
(1) Mixing 100g of ginseng raw material with 200g of acetic acid solution (acetic acid concentration is 1mol/L, pH is 2.38), performing acidification treatment at 45 ℃ for 20min to obtain an acidified solution, mixing the acidified solution with 1.5kg of methanol, performing reflux extraction at 110 ℃ for 3h, filtering the solution obtained by reflux extraction to obtain a filtrate, and performing rotary evaporation concentration on the filtrate at 50 ℃ to remove about 85% by volume of methanol to obtain 315g of concentrate as a first extract;
(2) Mixing 100g of activated clay, a ferric salt solution containing 0.6mol of ferric sulfate and 0.7mol of ferrous sulfate with a proper amount of sodium hydroxide solution under the anaerobic condition, carrying out contact reaction for 1h at the pH of 10 and the temperature of 80 ℃ to obtain a reaction solution, washing a solid substance obtained by filtering the reaction solution with water, and drying to obtain an auxiliary agent (the weight ratio of magnetic nano particles to activated clay is about 3:1); mixing the first extract with 1.6kg of water and 160g of auxiliary agent, reflux-extracting at 100deg.C for 1 hr to separate ginsenoside, and filtering with ultrafiltration membrane (filtering accuracy is 0.05 μm) to obtain second extract;
(3) Subjecting the second extract to silica gel column chromatography, eluting with 70 vol% methanol-water solution, collecting eluate, concentrating, and spray drying to obtain ginsenoside extract.
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Example 3
(1) Mixing 100g of ginseng raw material with 300g of acetic acid solution (acetic acid concentration is 4.5mol/L, pH is 2.05), performing acidification treatment at 30 ℃ for 30min to obtain an acidified solution, mixing the acidified solution with 2kg of ethanol, performing reflux extraction at 85 ℃ for 6h, filtering the solution obtained by reflux extraction to obtain a filtrate, and performing rotary evaporation concentration on the filtrate at 55 ℃ to remove about 90 vol% of ethanol to obtain 422g of concentrate as a first extract;
(2) Mixing 100g of activated clay, a ferric salt solution containing 0.85mol of ferric sulfate and 0.85mol of ferrous sulfate with a proper amount of sodium hydroxide solution under the anaerobic condition, carrying out contact reaction for 6 hours under the conditions of pH of 12 and temperature of 50 ℃ to obtain a reaction solution, washing a solid substance obtained by filtering the reaction solution with water, and drying to obtain an auxiliary agent (the weight ratio of magnetic nano particles to activated clay is about 4:1); mixing the first extract with 4kg of water and 400g of auxiliary agent, reflux-extracting at 85deg.C for 2 hr to separate ginsenoside, and filtering with ultrafiltration membrane (filtering accuracy is 0.05 μm) to obtain second extract;
(3) Subjecting the second extract to silica gel column chromatography, eluting with 80 vol% methanol-water solution, collecting eluate, concentrating, and spray drying to obtain ginsenoside extract.
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Example 4
Ginsenoside extraction was performed as in example 1, except that acetic acid was replaced with equal volume of citric acid (citric acid concentration 1g/L, pH is 2.8).
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Example 5
Ginsenoside extraction was performed as in example 1, except that acetic acid was replaced with an equal volume of hydrochloric acid (hydrochloric acid concentration of 0.01mol/L, pH of 2).
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Example 6
Extraction of ginsenoside was performed as in example 1, except that step (2) was replaced with:
(2) After mixing the first extract with 3.64kg of water and 364g of an auxiliary agent (81 g of activated clay and 283g of commercially available ferroferric oxide nanoparticles), reflux extraction was performed at 90 ℃ for 1.5 hours to separate ginsenoside, and filtration was performed by an ultrafiltration membrane (filtration accuracy is 0.05 μm) to obtain a second extract.
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Example 7
Extraction of ginsenoside was performed as in example 1, except that step (2) was replaced with:
(2) The first extract was mixed with 3.64kg of water and 364g of activated clay, and then subjected to reflux extraction at 90℃for 1.5 hours to separate ginsenoside, followed by ultrafiltration (filtration accuracy: 0.05 μm) to obtain a second extract.
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Example 8
Extraction of ginsenoside was performed as in example 1, except that step (2) was replaced with:
(2) The first extract was mixed with 3.64kg of water and 364g of commercially available ferroferric oxide nanoparticles, followed by reflux extraction at 90℃for 1.5 hours to separate ginsenoside, and filtration was performed by ultrafiltration membrane (filtration accuracy 0.05 μm) to obtain a second extract.
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Example 9
Extraction of ginsenoside was performed as in example 1, except that step (2) was replaced with:
(2) After mixing the first extract with 3.64kg of water and 364g of an auxiliary agent (81 g of activated clay and 283g of manganese dioxide nanoparticles purchased in the market), reflux extraction was performed at 90 ℃ for 1.5 hours to separate ginsenoside, and the second extract was obtained by ultrafiltration membrane filtration (filtration accuracy is 0.05 μm).
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Example 10
Extraction of ginsenoside was performed as in example 1, except that step (2) was replaced with:
(2) Mixing 100g of activated clay, a ferric salt solution containing 0.75mol of ferric sulfate and 0.75mol of ferrous sulfate with a proper amount of sodium hydroxide solution under the anaerobic condition, carrying out contact reaction for 4 hours under the conditions of pH 11 and temperature 65 ℃ to obtain a reaction solution, washing a solid substance obtained after filtering the reaction solution with water, and drying to obtain an auxiliary agent (the weight ratio of magnetic nano particles to activated clay is about 3.5:1); mixing the first extract with 3.64kg of water and 364g of auxiliary agent, reflux-extracting at 90deg.C for 1.5 hr to separate ginsenoside, and filtering with filter paper to obtain second extract.
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Example 11
Extraction of ginsenoside was performed as in example 1, except that step (3) was replaced with:
(3) And (3) passing the second extract through a nanofiltration membrane, concentrating the second extract until the solid content is 10% to obtain a concentrated solution, and performing spray drying on the concentrated solution to obtain the ginsenoside extract.
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Example 12
Extraction of ginsenoside was performed as in example 1, except that step (3) was replaced with:
(3) Subjecting the second extract to silica gel column chromatography, eluting with methanol, collecting eluate, concentrating, and spray drying to obtain ginsenoside extract.
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Comparative example 1
Extraction of ginsenoside was performed as in example 1, except that step (1) was replaced with:
(1) 100g of ginseng raw material, 400g of acetic acid solution (acetic acid concentration is 2mol/L, pH and is 2.23) and 1.8kg of ethanol are mixed, reflux extraction is carried out for 5 hours at the temperature of 100 ℃, the solution obtained by the reflux extraction is filtered to obtain filtrate, and the filtrate is subjected to rotary evaporation concentration at the temperature of 50 ℃ to remove about 90% by volume of ethanol, so that a concentrate is obtained as a first extract.
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Comparative example 2
Extraction of ginsenoside was performed as in example 1, except that step (1) was replaced with:
(1) 100g of ginseng raw material is mixed with 1.8kg of ethanol, reflux extraction is performed for 5 hours at a temperature of 100 ℃, a solution obtained by the reflux extraction is filtered to obtain a filtrate, the filtrate is subjected to rotary evaporation concentration at a temperature of 50 ℃ to remove about 90% by volume of ethanol, and 456g of concentrate is obtained as a first extract.
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Comparative example 3
Extraction of ginsenoside was performed as in example 1, except that step (2) was replaced with:
(2) The first extract was mixed with 3.64kg of a second extraction solvent and 364g of diatomaceous earth, and then subjected to reflux extraction at 90℃for 1.5 hours to separate ginsenoside, and filtered by an ultrafiltration membrane (filtration accuracy: 0.05 μm) to obtain a second extract.
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
Comparative example 4
Extraction of ginsenoside was performed as in example 1, except that step (2) was replaced with:
(2) The first extract was mixed with 3.64kg of water, 364g of an auxiliary agent (81 g of activated clay and 283g of diatomaceous earth), and then subjected to reflux extraction at 90℃for 1.5 hours to separate ginsenoside, and filtered by an ultrafiltration membrane (filtration accuracy: 0.05 μm) to obtain a second extract.
The total content of ginsenoside Re, rg1, rb1, ro, rc, rb2, rd in the ginsenoside extract was calculated, and the results are shown in Table 1.
TABLE 1
Numbering device Yield of ginsenoside (mg/g) Ginsenoside content (weight%)
Example 1 9.4 56.8
Example 2 8.8 57.1
Example 3 9.5 56.6
Example 4 8.1 54.5
Example 5 7.4 55.6
Example 6 8.3 54.2
Example 7 7.2 53.6
Example 8 7.5 53.8
Example 9 7.1 54.1
Example 10 7.2 53.9
Example 11 8.2 55.3
Example 12 8.4 55.6
Comparative example 1 6.8 52.1
Comparative example 2 5.1 49.5
Comparative example 3 6.1 53.1
Comparative example 4 6.3 53.3
As can be seen from Table 1, the method of the present invention can obtain higher yield and content of ginsenoside. Under the preferred scheme, the content and the yield can be further improved.
Test verification
1g of the ginsenoside extracts obtained in examples 1 to 12 and comparative examples 1 to 4 were dissolved in 20g of purified water, and the state was observed to obtain the ginsenoside extracts, respectively, in which the ginsenoside extracts obtained in examples 1 to 12 and comparative examples 1 to 4 were completely dissolved in water and a clear and transparent solution was formed; the respective solutions of ginsenoside extracts were subjected to accelerated stability test for 6 months, and the conditions were observed to obtain the respective solutions of ginsenoside extracts of examples 1 to 12 and comparative examples 3 to 4, which remained clear and transparent after 6 months, and were stable and not precipitated, and the respective solutions of ginsenoside extracts of comparative examples 1 to 2 were clouded.
Therefore, the ginsenoside extract obtained by the extraction method provided by the invention can avoid forming emulsion in a long-time state in a dissolved solution, and influence the clarity of the solution, so that the obtained ginsenoside extract has very good solution stability.
The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, a number of simple variants of the technical solution of the invention are possible, including combinations of the individual technical features in any other suitable way, which simple variants and combinations should likewise be regarded as being disclosed by the invention, all falling within the scope of protection of the invention.

Claims (10)

1. The extraction method of the ginsenoside is characterized by comprising the following steps:
(1) Mixing ginseng raw material with an acidic solution for acidizing treatment, and then mixing with a first extraction solvent for reflux extraction to obtain a first extraction solution;
(2) Mixing the first extract with a second extraction solvent and an auxiliary agent, performing reflux extraction II to separate ginsenoside, and performing solid-liquid separation I to obtain a second extract;
(3) Purifying the second extract, and drying to obtain ginsenoside extract;
wherein the conditions of the acidification treatment in step (1) include: the pH is 2-2.5, the temperature is 30-45 ℃ and the time is 20-30min; the first extraction solvent is selected from at least one of methanol, ethanol and water; in the step (2), the auxiliary agent is magnetic nano particles and/or activated clay, and the second extraction solvent is water; the purification treatment in step (3) comprises the following steps: subjecting the second extract to silica gel column chromatography, and collecting eluate; the eluent of the silica gel column chromatography is methanol-water solution.
2. The extraction method according to claim 1, wherein in the step (1), the ginseng raw material is prepared by drying and pulverizing ginseng;
the acidic solution is an organic acid solution.
3. The extraction method according to claim 1, wherein the acidic solution is an acetic acid solution.
4. The extraction method according to claim 1, wherein the first extraction solvent in step (1) is ethanol and/or methanol;
the weight ratio of the ginseng raw material to the first extraction solvent is 1:15-20.
5. The extraction method according to claim 1, wherein the conditions for reflux-extracting I in step (1) include: the temperature is 85-110 ℃ and the time is 3-6h.
6. The method according to any one of claims 1 to 5, wherein the step (2) further comprises concentrating the first extract under reduced pressure to remove the first extraction solvent before mixing the first extract with the second extraction solvent and the auxiliary agent.
7. The extraction method according to any one of claims 1 to 5, wherein the weight ratio of the first extraction solution, the second extraction solvent and the auxiliary agent in step (2) is 1:5-10:0.5-1.
8. The extraction method according to any one of claims 1 to 5, wherein the auxiliary agent is magnetic nanoparticles and activated clay;
the weight ratio of the magnetic nano particles to the activated clay is 3-4:1.
9. The method of extraction of claim 8, wherein the magnetic nanoparticles in step (2) are ferroferric oxide nanoparticles;
the preparation method of the auxiliary agent comprises the following steps: in the presence of activated clay, carrying out a contact reaction between ferric salt solution containing iron ions and ferrous ions and alkaline solution, and then carrying out solid-liquid separation II and washing;
the conditions of the contact reaction include: under the anaerobic condition, the pH value is 10-12, the temperature is 50-80 ℃ and the time is 1-6h.
10. The extraction method according to any one of claims 1 to 5, wherein in step (2), the conditions of the reflux extraction II include: the temperature is 85-100 ℃ and the time is 1-2h;
the solid-liquid separation I adopts ultrafiltration membrane filtration, and the filtration precision of the ultrafiltration membrane is 0.01-0.1 mu m.
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