CN106086146A - A kind of Preparation method and use of refined precious rare ginsenoside - Google Patents

A kind of Preparation method and use of refined precious rare ginsenoside Download PDF

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Publication number
CN106086146A
CN106086146A CN201610501690.8A CN201610501690A CN106086146A CN 106086146 A CN106086146 A CN 106086146A CN 201610501690 A CN201610501690 A CN 201610501690A CN 106086146 A CN106086146 A CN 106086146A
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preparation
ginsenoside
precious rare
saponin
rare ginsenoside
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赵兴增
王相云
单宇
李龙柱
冯煦
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Institute of Botany of CAS
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    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P33/00Preparation of steroids
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P33/00Preparation of steroids
    • C12P33/20Preparation of steroids containing heterocyclic rings
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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    • Y02PCLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00Technologies relating to chemical industry
    • Y02P20/50Improvements relating to the production of bulk chemicals
    • Y02P20/54Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids

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Abstract

The present invention relates to field of natural medicinal chemistry, disclose one and utilize panax species extract to generate precious rare ginsenoside Preparation method and use.Comprise the following steps: use solvent refluxing to extract by after panax species section or pulverizing, after filtering extracting solution, extracting solution is concentrated to give extractum;Extractum use de-agriculture residual and removing heavy-metal process;Producing panaxdiols saponin, concentrate drying becomes sterilizing after powder, is subsequently cooled to uniform temperature;Sterilized panaxdiols saponin powder is mixed with fermented bacterium, regulates moisture with aseptic liquid nutrient medium and distilled water, fully seal after mixing, be placed in constant incubator fermentation and obtain panaxdiols saponin tunning.Present invention process is simple, easy to operate, and a large amount of acquisitions are rare and have antitumor and antidepressant activity material preciousness rare ginsenoside.

Description

A kind of Preparation method and use of refined precious rare ginsenoside
Technical field
The invention belongs to Natural Medicine Chemistry field of bioconversion, be specifically related to one and utilize panax species fermentation to generate The preparation method of precious rare ginsenoside and antitumor, raising immunity, combined radio chemotherapy drug-enhancer attenuation and antidepressant The application of aspect.
Background technology
Being mainly composed of ginsenoside in panax species extract, ginsenoside is divided into three kinds according to the difference of parent nucleus Type: protopanaxadiol-type, Protopanaxatriol's type and oleanolic acid type.
Panaxdiols saponin is the general designation of the serial ginsenoside with protopanoxadiol as parent nucleus, is in panax species Antitumor, the raising the most significant material of the physiologically active such as immunity and antidepressant.
Modern pharmacological research finds, the anti-tumor activity of ginsenoside and the binding site of ginsenoside's molecular side chain glycosyl Relevant with quantity, and anti-tumor activity increases along with the minimizing of glycosyl quantity, and the Panaxadiol saponin fermentation of short sugar chain is produced Thing has the strongest antitumor and antidepressant activity.
Precious rare short sugar chain ginsenoside is that content is extremely low in panax species, but has a class of notable physiologically active Saponin, mainly: protopanoxadiol, ginsenoside Rg3, GF2, ginsenoside Rh2 and Ginsenoside compound K.Pharmacology is real Checking is real, and precious rare short sugar chain ginsenoside has suppression multiple growth of cancer cells, antidepressant, activation chloride channel, presses down Many work such as sodium channel depolarization, suppression human embryo kidney (HEK), the extension of HEK-293 cell and H. pylori bacteria growing that system is activated Property.
Summary of the invention
It is an object of the invention to provide a kind of preparation side utilizing panax species fermentation to generate precious rare ginsenoside Method and antitumor, raising immunity, combined radio chemotherapy drug-enhancer attenuation and the application of anti-depression aspect.Present invention process letter Single, easy to operate, product drying is easy, it is most important that the biology realizing panaxdiols saponin during strain fermentation turns Change, obtain antitumor and antidepressant activity material preciousness rare ginsenoside.
The purpose of the present invention is by realizing by the following technical solutions
The preparation method of a kind of refined precious rare ginsenoside, comprises the following steps:
A. by panax species section or pulverizing;
B. use organic extraction solvent reflux, extract, filter, filtrate reduced in volume, obtain panax species extractum;
C., panax species extractum use de-agriculture residual and removing heavy-metal technique;
D. panax species extractum is purified process, prepares panaxdiols saponin, and be dried to powder;
E. by panaxdiols saponin powder low temperature sterilization, it is subsequently cooled to uniform temperature;
F. being mixed with fermented bacterium by sterilized panaxdiols saponin powder, strain addition is 1%-100%, uses sterile liquid Body culture medium and distilled water regulation moisture, fully seal after mixing, being placed in constant incubator fermentation and obtaining panoxadiol and organize soap Glycosides tunning.
Further, described panax species is Radix Ginseng, Radix Panacis Quinquefolii or Radix Notoginseng.
Further, described organic extraction solvent is water, ethanol or methanol.
Further, the described residual technique of de-agriculture is supercritical extraction, removing heavy-metal technique be chelating resin D751 or D403。
Further, described panaxdiols saponin technique of preparing is column chromatography, and chromatographic material is macroporous resin.
Further, in described step e, panax species extract powder sterilising temp is 40-100 DEG C.
Further, in described f step, the moisture of fermentation system is 30%-90%, and fermentation time is 4-24h, fermentation temperature Degree is for 20-80 DEG C.
Further, in described sweat, the C-3 of fermented bacterium Polysaccharides chain panax species extract saponin and The glycosyl of C-20 position, is converted into the part anti-tumor active substance panaxdiols saponin tunning without sugar chain.
Further, the refined precious rare ginsenoside preparation method of described one, it is characterised in that described fermented In journey, panax species extract fermented bacterium is from fungus, antibacterial or actinomycetes such as bread mold Rhizopus sp strain.
Further, described panaxdiols saponin tunning comprises protopanoxadiol, ginsenoside Rg3, Radix Ginseng soap Glycosides F2, ginsenoside Rh2 and Ginsenoside compound K etc.
The strain fermentation of ginsenoside of the present invention is as nutrient matrix using panax species extract, by the biochemistry of microorganism Course of reaction carries out structural modification to ginsenoside, produces new active component.The method technique is simple, easy to operate, product It is dried easily, with low cost, there is high specificity, high efficiency, environmental friendliness, be suitable for the features such as industry amplification, it is achieved that precious The preparation of rare ginsenoside.
Accompanying drawing illustrates:
Can report as APPENDIX MATERIALSThe using figure below.
Fig. 1, precious rare ginsenoside preparation flow figure
Detailed description of the invention:
Understand the present invention in order to clearer, be described in detail these embodiments below by specific embodiment further and only use In the present invention is further described, two are not used in restriction the scope of the present invention.
Embodiment 1
Panax species Radix Ginseng is cut into slices, and uses 70% alcohol reflux, filters, filtrate reduced in volume, obtains Radix Ginseng extractum, to people It is residual that ginseng extractum uses supercritical extraction to take off agriculture, and i.e. selecting 10% ethanol is 0.5:1 as the ratio of entrainer, entrainer and raw material, super The extraction of critical extraction is 30MPa, extraction 60 degrees Celsius, uses supercritical extracting equipment, and CO2 recycles, flow rate set 5kg/h, average dose is 15kg.Using chelating resin to take off agriculture Radix Ginseng extractum residual, 500g extractum is diluted with water to 5L, stirring, Standing 12h, take supernatant and cross D751 resin, 95% ethanol elution, merge eluent, vacuum rotating is concentrated into without alcohol taste, adds water to 2L, through macroporous resin D101 adsorb, 40-50% ethanol elution stream part, concentrated after obtain dried powder, 121 DEG C of sterilizings 15min, is cooled to room temperature, low temperature sterilization, and sterilising temp is 40-100 DEG C, is then cooled to room temperature;By the Radix Ginseng after 10g sterilizing Genus plants extract powder mixes with 2.2g strain;Add fluid medium and distilled water regulation moisture, the moisture of fermentation system For 30%-90%;Fully seal after mixing, be placed in 30 DEG C of constant incubators fermentation 8d and obtain precious rare ginsenoside.
Embodiment 2
Panax species pseudo-ginseng root slices, uses 80% alcohol reflux, filters, filtrate reduced in volume, obtains Radix Ginseng extractum, to people It is residual that ginseng extractum uses supercritical extraction to take off agriculture, and i.e. selecting 15% ethanol is 0.8:1 as the ratio of entrainer, entrainer and raw material, super The extraction of critical extraction is 30MPa, extraction 60 degrees Celsius, uses supercritical extracting equipment, and CO2 recycles, flow rate set 5kg/h, average dose is 15kg.Using chelating resin to take off agriculture Radix Ginseng extractum residual, 500g extractum is diluted with water to 5L, stirring, Standing 12h, take supernatant and cross D403 resin, 95% ethanol elution, merge eluent, vacuum rotating is concentrated into without alcohol taste, adds water to 2L, through macroporous resin D101 adsorb, 40-50% ethanol elution stream part, concentrated after obtain dried powder, low temperature sterilization, sterilizing temperature Degree, for 40-100 DEG C, is then cooled to room temperature;Configuration fluid medium, 121 DEG C of sterilizing 15min, it is cooled to room temperature;10g is gone out Panax species extract powder after bacterium mixes with 2.2g strain;Add fluid medium and distilled water regulation moisture, fermentation The moisture of system is 30%-90%;Fully seal after mixing, be placed in 40 DEG C of constant incubators fermentation 10d and obtain precious rare people Ginseng saponin;
Embodiment 3
Panax species Radix Panacis Quinquefolii is cut into slices, and uses 75% alcohol reflux, filters, filtrate reduced in volume, obtains Radix Ginseng extractum, right It is residual that Radix Ginseng extractum uses supercritical extraction to take off agriculture, and i.e. selecting 10% ethanol is 0.8:1 as the ratio of entrainer, entrainer and raw material, The extraction of supercritical extraction is 30MPa, extraction 60 degrees Celsius, uses supercritical extracting equipment, and CO2 recycles, flow rate set 5kg/h, average dose is 15kg.Using chelating resin to take off agriculture Radix Ginseng extractum residual, 500g extractum is diluted with water to 5L, stirring, Standing 12h, take supernatant and cross D751 resin, 95% ethanol elution, merge eluent, vacuum rotating is concentrated into without alcohol taste, adds water to 2L, through macroporous resin D101 adsorb, 40-50% ethanol elution stream part, concentrated after obtain dried powder, low temperature sterilization, sterilizing temperature Degree, for 40-100 DEG C, is then cooled to room temperature;Configuration fluid medium, 121 DEG C of sterilizing 15min, it is cooled to room temperature by 8g sterilizing After glycol saponins powder and 2.5g strain mix;Add fluid medium and distilled water regulation moisture, the water of fermentation system It is divided into 30%-90%;Fully seal after mixing, be placed in 50 DEG C of constant incubators fermentation 12d and obtain precious rare ginsenoside.

Claims (12)

1. a Preparation method and use for refined precious rare ginsenoside, comprises the following steps:
A, by panax species section or pulverize;
B, use organic extraction solvent solution reflux, extract, filter, filtrate reduced in volume, obtain panax species extractum;
C, panax species extractum use de-agriculture residual and removing heavy-metal technique;
D, panax species extractum is purified process, prepares panaxdiols saponin, and be dried to powder;
E, by panaxdiols saponin powder low temperature sterilization, be subsequently cooled to uniform temperature;
F, sterilized panaxdiols saponin powder and fermented bacterium being mixed, strain addition is 1%-100%, with aseptic Fluid medium and distilled water regulation moisture, fully seal after mixing, be placed in constant incubator fermentation and obtain panoxadiol's group The refined precious rare ginsenoside of saponin tunning.
The preparation method of a kind of refined precious rare ginsenoside the most according to claim 1, it is characterised in that described Panax species is Radix Ginseng, the mixture of one or more of Radix Panacis Quinquefolii or Radix Notoginseng.
The preparation method of a kind of refined precious rare ginsenoside the most according to claim 1, it is characterised in that described Organic extraction solvent is water, ethanol or methanol.
The preparation method of a kind of refined precious rare ginsenoside the most according to claim 1, it is characterised in that described The de-residual technique of agriculture is supercritical extraction, and removing heavy-metal technique is chelating resin D751 or D403.
The preparation method of a kind of refined precious rare ginsenoside the most according to claim 1, it is characterised in that described Preparing panaxdiols saponin purifying process is column chromatography, and chromatography material is macroporous resin.
The preparation method of a kind of refined precious rare ginsenoside the most according to claim 1, it is characterised in that described e In step, panax species extract powder sterilising temp is 40-100 DEG C.
The preparation method of a kind of refined precious rare ginsenoside the most according to claim 1, it is characterised in that described f In step, the moisture of fermentation system is 30%-90%, and fermentation time is 4-24h, and fermentation temperature is 20-80 DEG C.
The preparation method of a kind of refined precious rare ginsenoside the most according to claim 1, it is characterised in that described During ferment, the sugar of C-3 and the C-20 position of fermented bacterium Polysaccharides chain panax species extract saponin, converted product is with nothing Sugar chain saponin protopanoxadiol is main, with short sugar chain saponin ginsenoside Rg3, GF2, ginsenoside Rh2, Radix Ginseng soap Glycosides CK is an auxiliary class mixture.
The preparation method of a kind of refined precious rare ginsenoside the most according to claim 1, it is characterised in that described During ferment, panax species extract fermented bacterium is from fungus, antibacterial or actinomycetes such as bread mold Rhizopus sp strain.
The preparation method of a kind of refined precious rare ginsenoside the most according to claim 1, it is characterised in that described essence The precious rare ginsenoside of system is mainly protopanoxadiol, ginsenoside Rg3, GF2, ginsenoside Rh2, Radix Ginseng soap Glycosides CK etc..
The preparation method of 11. a kind of refined precious rare ginsenosides according to claim 1, it is characterised in that described use Way mainly antitumor, raising immunity, combined radiotherapy or chemotherapeutics can efficacy enhancing and toxicity reducing and antidepressants.
The preparation method of 12. a kind of refined precious rare ginsenosides according to claim 1, it is characterised in that described use On the way, food additive, health product and medicine it are mainly used in.
CN201610501690.8A 2016-06-30 2016-06-30 A kind of Preparation method and use of refined precious rare ginsenoside Pending CN106086146A (en)

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CN106900856A (en) * 2017-03-07 2017-06-30 吉林大学 The balloonflower root fermented ginseng Yoghourt and its production method of a kind of green
CN109646471A (en) * 2019-01-22 2019-04-19 名臣健康用品股份有限公司 A kind of activation notoginsenoside rich in rare ginsenoside is in the application prepared in oral care product
CN110156864A (en) * 2019-06-02 2019-08-23 上海纳米技术及应用国家工程研究中心有限公司 A kind of preparation method of the black ginseng Rg3 of supercritical carbon dioxide extracting
CN110283871A (en) * 2019-07-10 2019-09-27 吉林农业大学 A method of polysaccharide and rare saponin(e are produced using compound fungus fermented ginseng
CN112791110A (en) * 2021-04-12 2021-05-14 南京乔康生物科技有限公司 Preparation method of ginseng root extract with low pesticide residue
CN114404465A (en) * 2022-01-06 2022-04-29 解慧勇 A black ginseng paste for treating tumor and its preparation method

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106900856A (en) * 2017-03-07 2017-06-30 吉林大学 The balloonflower root fermented ginseng Yoghourt and its production method of a kind of green
CN109646471A (en) * 2019-01-22 2019-04-19 名臣健康用品股份有限公司 A kind of activation notoginsenoside rich in rare ginsenoside is in the application prepared in oral care product
CN110156864A (en) * 2019-06-02 2019-08-23 上海纳米技术及应用国家工程研究中心有限公司 A kind of preparation method of the black ginseng Rg3 of supercritical carbon dioxide extracting
CN110283871A (en) * 2019-07-10 2019-09-27 吉林农业大学 A method of polysaccharide and rare saponin(e are produced using compound fungus fermented ginseng
CN112791110A (en) * 2021-04-12 2021-05-14 南京乔康生物科技有限公司 Preparation method of ginseng root extract with low pesticide residue
CN114404465A (en) * 2022-01-06 2022-04-29 解慧勇 A black ginseng paste for treating tumor and its preparation method

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Application publication date: 20161109