CN104398549A - Method for preparation of total saponins of panax ginseng by high-voltage pulse electric field assisted enzyme hydrolysis - Google Patents
Method for preparation of total saponins of panax ginseng by high-voltage pulse electric field assisted enzyme hydrolysis Download PDFInfo
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- CN104398549A CN104398549A CN201410639581.3A CN201410639581A CN104398549A CN 104398549 A CN104398549 A CN 104398549A CN 201410639581 A CN201410639581 A CN 201410639581A CN 104398549 A CN104398549 A CN 104398549A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61K2236/30—Extraction of the material
- 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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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Abstract
The invention discloses a method for preparation of total saponins of panax ginseng by high-voltage pulse electric field assisted enzyme hydrolysis, and the method is as follows: taking 3g of panax ginseng dry powder, adding 50mL of distilled water; adding 1-3% beta-glucosidase liquid; performing enzyme hydrolysis in high-voltage pulse electric field with the electric field strength of 5-25kV/cm, wherein processed pulse number is 6-12; after the completion of the reaction, heating at 100 DEG C for 5min for passivating; using water saturated butanol for extraction reaction liquid, centrifugally separating, taking supernatant; performing vacuum rotary evaporation on extract of the total saponins of panax ginseng at 55 DEG, and dissolving a sample in methanol. Due to the introduction of high-voltage pulse electric field in the enzyme hydrolysis reaction process, the enzyme preparation activity is improved, the enzyme hydrolysis reaction is accelerated, and dissolution of the total saponin in the panax ginseng can be fast and efficiently promoted.
Description
Technical field
The present invention relates to a kind of method preparing Radix Ginseng total saponins, particularly relate to the method that Radix Ginseng total saponins is prepared in the hydrolysis of a kind of high-pressure pulse electric auxiliary enzymes.
Background technology
Radix Ginseng (Panax ginseng C.A.Meyer) is araliaceae ginseng plant, is traditional rare Chinese medicine.Radix Ginseng has strongly invigorating primordial QI, invigorating the spleen to benefit the lung, promoting the production of body fluid to quench thirst, effects such as increasing intelligence of calming the nerves.Containing Multiple components active component such as ginsenoside, volatile oil, organic acid, esters and polysaccharides in Radix Ginseng, wherein ginsenoside is the main active of Radix Ginseng.At present, from Radix Ginseng, at least isolated more than 40 and planted ginsenoside monomer.
Ginsenoside to be connected with glycosyl the plant sugar glycoside material formed by aglycon, is extensively present in the root of Radix Ginseng, stem, leaf, Hua Zhong.Ginsenoside all belongs to triterpene saponin, can be divided into three types: oleanolic acid type saponin is as Ro by its sapogenin structure difference; Protopanaxadiol-type's saponin; Protopanaxatriol's type saponin.Rear two classes all belong to the tetracyclic triterpene saponin of dammarane type, are one of main active of Radix Ginseng.
At present, the traditional extraction process of ginsenoside comprises organic solvent extraction, heating reflux method, soxhlet extraction and steam distillation etc.But there is the problems such as extracting cycle is long, solvent load is high, loss of effective components is large, energy consumption is high.
Summary of the invention
The object of the invention is to propose a kind of high-pressure pulse electric auxiliary enzymes and be hydrolyzed the method preparing Radix Ginseng total saponins, high to overcome Extracting temperature in current traditional extraction process, the shortcomings such as extraction time is long.
The present invention realizes by the following technical solutions:
A method for Radix Ginseng total saponins is prepared in the hydrolysis of high-pressure pulse electric auxiliary enzymes, comprises the following steps:
1) get Radix Ginseng dried powder, add distilled water,
2) enzyme preparation is added again,
3) enzyme hydrolysis under high-voltage pulse electric field action;
4) treat that above-mentioned reaction completes, add thermal inactivation,
5) add organic solvent extraction, centrifugalize, get supernatant,
6) concentrated extracting solution, sample dissolution is in methanol.
An improvement project for the method for Radix Ginseng total saponins is prepared in the hydrolysis of high-pressure pulse electric auxiliary enzymes, described step 1) specifically refer to: get 3g Radix Ginseng dried powder, add 50mL distilled water.
The improvement project of the method for Radix Ginseng total saponins, described step 2 are prepared in the hydrolysis of high-pressure pulse electric auxiliary enzymes) in the enzyme preparation that adds be beta-glucosidase.
The improvement project of the method for Radix Ginseng total saponins, described step 2 are prepared in the hydrolysis of high-pressure pulse electric auxiliary enzymes) in the addition of enzyme preparation be 1-3%.
The improvement project of the method for Radix Ginseng total saponins, described step 3 are prepared in the hydrolysis of high-pressure pulse electric auxiliary enzymes) electric field intensity of mesohigh impulse electric field is 5-25kV/cm, the umber of pulse of process is 6-12.
The improvement project of the method for Radix Ginseng total saponins, described step 4 are prepared in the hydrolysis of high-pressure pulse electric auxiliary enzymes) specifically referring to will by described step 3) solution 100 DEG C after enzymolysis heats 5min, makes its passivation.
The improvement project of the method for Radix Ginseng total saponins, described step 5 are prepared in the hydrolysis of high-pressure pulse electric auxiliary enzymes) in the organic solvent that adds be n-butyl alcohol, and use water saturation solution.
An improvement project for the method for Radix Ginseng total saponins is prepared in the hydrolysis of high-pressure pulse electric auxiliary enzymes, step 6) middle concentrated extracting solution, specifically refer to Radix Ginseng total saponins extracting solution in 55 DEG C of rotary evaporation in vacuo.
Compared with prior art, the present invention has the following advantages:
This method, owing to introducing high-pressure pulse electric in enzyme digestion reaction process, improves enzyme preparation active, accelerates enzyme digestion reaction speed, promotes total saponins material stripping in Radix Ginseng fast and efficiently.
The present invention utilizes the hydrolysis of high-voltage pulse electric field technology auxiliary commerce enzyme preparation to prepare Radix Ginseng total saponins, has without the need to heating, response speed is fast, extraction ratio is high, low cost and other advantages.
The inventive method has mild condition, does not destroy Saponin structure, extraction time is short, process conditions are simple, sample purity advantages of higher, and the enzyme adopted is cheap commercialization enzyme.
Detailed description of the invention
In conjunction with below the embodiment that provides the inventive method is described in further detail.
The method of Radix Ginseng total saponins is prepared in the hydrolysis of high-pressure pulse electric auxiliary enzymes: get a 3g Radix Ginseng dried powder, add 50mL distilled water; Add the beta-glucosidase liquid of 1-3% again; Enzyme hydrolysis under high-voltage pulse electric field action, electric field intensity is 5-25kV/cm, and the umber of pulse of process is 6-12; Treat that above-mentioned reaction completes, 100 DEG C of heating 5min, make its passivation; With water-saturated n-butanol extractive reaction liquid, centrifugalize, gets supernatant; By Radix Ginseng total saponins extracting solution in 55 DEG C of rotary evaporation in vacuo, sample dissolution is in methanol.
Embodiment 1
Radix Ginseng total saponins is prepared in high-pressure pulse electric auxiliary enzymes hydrolysis of the present invention, get 3g Radix Ginseng powder sample, add 50mL distilled water, magnetic stirrer is even, by glacial acetic acid adjust ph to 5.0, adds 1% beta-glucosidase liquid, concentration of substrate is 0.5mg/mL, enzymolysis under PEF effect, electric field intensity is 10kV/cm, and umber of pulse is 6; With water-saturated n-butanol extractive reaction liquid, centrifugalize, supernatant volatilization is except desolventizing, and the yield through efficient liquid phase chromatographic analysis ginsenoside Rd is 2.36%.
Embodiment 2
Radix Ginseng total saponins is prepared in high-pressure pulse electric auxiliary enzymes hydrolysis of the present invention, get 3g Radix Ginseng powder sample, add 50mL distilled water, magnetic stirrer is even, by glacial acetic acid adjust ph to 5.0, adds 1% beta-glucosidase liquid, concentration of substrate is 1.0mg/mL, enzymolysis under PEF effect, electric field intensity is 10kV/cm, and umber of pulse is 6; With water-saturated n-butanol extractive reaction liquid, centrifugalize, supernatant volatilization is except desolventizing, and the yield through efficient liquid phase chromatographic analysis ginsenoside Rd is 3.43%.
Embodiment 3
Radix Ginseng total saponins is prepared in high-pressure pulse electric auxiliary enzymes hydrolysis of the present invention, get 3g Radix Ginseng powder sample, add 50mL distilled water, magnetic stirrer is even, by glacial acetic acid adjust ph to 5.0, adds 2% beta-glucosidase liquid, concentration of substrate is 1.0mg/mL, enzymolysis under PEF effect, electric field intensity is 10kV/cm, and umber of pulse is 8.With water-saturated n-butanol extractive reaction liquid, centrifugalize, supernatant volatilization is except desolventizing, and the yield through efficient liquid phase chromatographic analysis ginsenoside Rd is 3.52%.
Embodiment 4
Radix Ginseng total saponins is prepared in high-pressure pulse electric auxiliary enzymes hydrolysis of the present invention, get 3g Radix Ginseng powder sample, add 50mL distilled water, magnetic stirrer is even, by glacial acetic acid adjust ph to 5.0, adds 1% beta-glucosidase liquid, concentration of substrate is 1.0mg/mL, enzymolysis under PEF effect, electric field intensity is 5kV/cm, and umber of pulse is 6.With water-saturated n-butanol extractive reaction liquid, centrifugalize, supernatant volatilization is except desolventizing, and the yield through efficient liquid phase chromatographic analysis ginsenoside Rd is 2.18%.
Embodiment 5
Radix Ginseng total saponins is prepared in high-pressure pulse electric auxiliary enzymes hydrolysis of the present invention, get 3g Radix Ginseng powder sample, add 50mL distilled water, magnetic stirrer is even, by glacial acetic acid adjust ph to 5.0, adds 1% beta-glucosidase liquid, concentration of substrate is 1.0mg/mL, enzymolysis under PEF effect, electric field intensity is 20kV/cm, and umber of pulse is 6.With water-saturated n-butanol extractive reaction liquid, centrifugalize, supernatant volatilization is except desolventizing, and the yield through efficient liquid phase chromatographic analysis ginsenoside Rd is 3.38%.
Embodiment 6
Radix Ginseng total saponins is prepared in high-pressure pulse electric auxiliary enzymes hydrolysis of the present invention, get 3g Radix Ginseng powder sample, add 50mL distilled water, magnetic stirrer is even, by glacial acetic acid adjust ph to 5.0, adds 1% beta-glucosidase liquid, concentration of substrate is 1.0mg/mL, enzymolysis under PEF effect, electric field intensity is 10kV/cm, and umber of pulse is 12.With water-saturated n-butanol extractive reaction liquid, centrifugalize, supernatant volatilization is except desolventizing, and the yield through efficient liquid phase chromatographic analysis ginsenoside Rd is 3.94%.
Embodiment 7
Radix Ginseng total saponins is prepared in high-pressure pulse electric auxiliary enzymes hydrolysis of the present invention, get 3g Radix Ginseng powder sample, add 50mL distilled water, magnetic stirrer is even, by glacial acetic acid adjust ph to 5.0, adds 1% beta-glucosidase liquid, concentration of substrate is 1.0mg/mL, enzymolysis under PEF effect, electric field intensity is 25kV/cm, and umber of pulse is 6.With water-saturated n-butanol extractive reaction liquid, centrifugalize, supernatant volatilization is except desolventizing, and the yield through efficient liquid phase chromatographic analysis ginsenoside Rd is 3.06%.
Embodiment 8
Radix Ginseng total saponins is prepared in high-pressure pulse electric auxiliary enzymes hydrolysis of the present invention, get 3g Radix Ginseng powder sample, add 50mL distilled water, magnetic stirrer is even, by glacial acetic acid adjust ph to 5.0, adds 3% beta-glucosidase liquid, concentration of substrate is 1.0mg/mL, enzymolysis under PEF effect, electric field intensity is 10kV/cm, and umber of pulse is 8.With water-saturated n-butanol extractive reaction liquid, centrifugalize, supernatant volatilization is except desolventizing, and the yield through efficient liquid phase chromatographic analysis ginsenoside Rd is 3.55%.
Claims (8)
1. a method for Radix Ginseng total saponins is prepared in the hydrolysis of high-pressure pulse electric auxiliary enzymes, it is characterized in that, comprises the following steps:
1) get Radix Ginseng dried powder, add distilled water,
2) enzyme preparation is added again,
3) enzyme hydrolysis under high-voltage pulse electric field action;
4) treat that above-mentioned reaction completes, add thermal inactivation,
5) add organic solvent extraction, centrifugalize, get supernatant,
6) concentrated extracting solution, sample dissolution is in methanol.
2. the method for Radix Ginseng total saponins is prepared in a kind of high-pressure pulse electric auxiliary enzymes hydrolysis according to claim 1, it is characterized in that, described step 1) specifically refer to: get 3g Radix Ginseng dried powder, add 50mL distilled water.
3. the method for Radix Ginseng total saponins is prepared in a kind of high-pressure pulse electric auxiliary enzymes hydrolysis according to claim 1, it is characterized in that, described step 2) in the enzyme preparation that adds be beta-glucosidase.
4. the method for Radix Ginseng total saponins is prepared in a kind of high-pressure pulse electric auxiliary enzymes hydrolysis according to claim 1, it is characterized in that, described step 2) in the addition of enzyme preparation be 1-3%.
5. the method for Radix Ginseng total saponins is prepared in a kind of high-pressure pulse electric auxiliary enzymes hydrolysis according to claim 1, it is characterized in that, described step 3) electric field intensity of mesohigh impulse electric field is 5-25kV/cm, the umber of pulse of process is 6-12.
6. the method for Radix Ginseng total saponins is prepared in a kind of high-pressure pulse electric auxiliary enzymes hydrolysis according to claim 1, it is characterized in that, described step 4) specifically referring to will by described step 3) solution 100 DEG C after enzymolysis heats 5min, makes its passivation.
7. the method for Radix Ginseng total saponins is prepared in a kind of high-pressure pulse electric auxiliary enzymes hydrolysis according to claim 1, it is characterized in that, described step 5) in the organic solvent that adds be n-butyl alcohol, and use water saturation solution.
8. the method for Radix Ginseng total saponins is prepared in a kind of high-pressure pulse electric auxiliary enzymes hydrolysis according to claim 1, it is characterized in that, step 6) middle concentrated extracting solution, specifically refer to Radix Ginseng total saponins extracting solution in 55 DEG C of rotary evaporation in vacuo.
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Cited By (5)
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| CN105255962A (en) * | 2015-10-23 | 2016-01-20 | 齐齐哈尔医学院 | Method for extracting blood sugar-reducing panaxan by enzyme method and application of obtained panaxan |
| CN106551967A (en) * | 2015-09-28 | 2017-04-05 | 吴景太 | A kind of ginsenoside capsule |
| TWI614062B (en) * | 2015-12-30 | 2018-02-11 | 宣美國際股份有限公司 | Hydrolyzate extraction method |
| CN110583939A (en) * | 2019-10-22 | 2019-12-20 | 成都市农林科学院 | Rose clarified juice and rose low-sugar yogurt based on enzymolysis and preparation method thereof |
| CN113005157A (en) * | 2021-03-29 | 2021-06-22 | 吉林大学 | Preparation method of soybean isoflavone aglycone |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106551967A (en) * | 2015-09-28 | 2017-04-05 | 吴景太 | A kind of ginsenoside capsule |
| CN105255962A (en) * | 2015-10-23 | 2016-01-20 | 齐齐哈尔医学院 | Method for extracting blood sugar-reducing panaxan by enzyme method and application of obtained panaxan |
| TWI614062B (en) * | 2015-12-30 | 2018-02-11 | 宣美國際股份有限公司 | Hydrolyzate extraction method |
| CN110583939A (en) * | 2019-10-22 | 2019-12-20 | 成都市农林科学院 | Rose clarified juice and rose low-sugar yogurt based on enzymolysis and preparation method thereof |
| CN110583939B (en) * | 2019-10-22 | 2022-06-17 | 成都市农林科学院 | Rose clarified juice and rose low-sugar yogurt based on enzymolysis and preparation method thereof |
| CN113005157A (en) * | 2021-03-29 | 2021-06-22 | 吉林大学 | Preparation method of soybean isoflavone aglycone |
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