CN106074629A - The method that molecular engram extracts Ganoderma triterpenoids with ultrasonic coupling - Google Patents
The method that molecular engram extracts Ganoderma triterpenoids with ultrasonic coupling Download PDFInfo
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- CN106074629A CN106074629A CN201610500990.4A CN201610500990A CN106074629A CN 106074629 A CN106074629 A CN 106074629A CN 201610500990 A CN201610500990 A CN 201610500990A CN 106074629 A CN106074629 A CN 106074629A
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- molecular engram
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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
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
- A61K36/074—Ganoderma
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- 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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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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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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- 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
- A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract
The invention discloses a kind of method that molecular engram extracts Ganoderma triterpenoids with ultrasonic coupling, relate to the extraction and separation technology field of natural product.The present invention utilizes molecular imprinting using target molecule as template molecule, select function monomer matched on structure of functional groups, allow and covalent effect or noncovalent interaction occur between the two, add cross-linking agent by thermal polymerization or photopolymerization, polyreaction is produced under the induction of the porogen of initiator, thus formation i.e. molecularly imprinted polymer (MIPs), subsequently, utilize either physically or chemically, target molecule is eluted out from polymeric inner, once obtain the principle with identical with target molecule shape and that position of functional group is certain memory hole, without using organic solvent to extract for a long time, the extraction process time solving current Ganoderma triterpenoids is long, power consumption height, active component extract yield is low, residual machine solvent the problem affecting health.
Description
Technical field
The present invention relates to the extraction and separation technology field of natural product, especially a kind of molecular engram extracts with ultrasonic coupling
The method of Ganoderma triterpenoids.
Background technology
Ganoderma is On Polyporaceae Ganoderma also known as Lin Zhongling, fine jade precious (formal name used at school: Ganoderma Lucidum Karst)
Sporophore.Ganoderma contains multiple chemical composition, and its main active has polysaccharide and peptide polysaccharide class, alkaloids and nucleotide
Class, amino acid and protein class, triterpenes, trace element.Wherein ganoderma lucidum triterpene compounds has hepatoprotective effect, antitumor is made
Close with, AntiHIV1 RT activity 1 and HIV2 proteinase activity, suppression histamine release, Angiotensin-converting enzyme inhibition (ACE), suppression cholesterol
One-tenth, paroxysmal pain effect, the impact on platelet aggregation, the inhibitory action to albumen Acacia farnesiana Willd. transesterify enzyme, antioxidation etc..Mesh
Before, general employing organic solvent method extracts Ganoderma triterpenoids, it usually needs longer extraction time also consumes substantial amounts of Extraction solvent,
The use of the most a large amount of organic solvents can bring adverse influence to human body and environment；Further, since Ganoderma sporophore is by fiber
The main components such as element, hemicellulose and lignin are constituted, close structure, and traditional extraction time length, power consumption are high, active component
Extract yield is low.
Molecular imprinting is the focus of research at present, and it has structure effect precordainment, the feature of specific recognition, is a kind of
Relatively there is the technology of industrialization value.This technology is usually target molecule as template molecule, selects in sense unity
Function monomer matched on structure, allows and covalent effect or noncovalent interaction occurs between the two, adds cross-linking agent by heat
Polymerization or photopolymerization, produce polyreaction under the induction of the porogen of initiator, thus form the height being enclosed with target molecule
Polymers, i.e. molecularly imprinted polymer (MIPs), subsequently, utilize physics or chemical method, by target molecule from polymeric inner
It is eluted out, once obtains and there is identical with target molecule shape and that position of functional group is certain memory hole.
Summary of the invention
The present invention provides a kind of method that molecular engram extracts Ganoderma triterpenoids with ultrasonic coupling, and it can solve current Ganoderma
The extraction process time length of triterpene, power consumption is high, active component extract yield is low, residual machine solvent affect asking of health
Topic.
In order to solve the problems referred to above, the technical solution adopted in the present invention is: this molecular engram extracts with ultrasonic coupling
The method of Ganoderma triterpenoids, is using REISH as extracting raw material, using water as extracting medium, utilizes molecular engram material to adsorb
Ganoderma triterpenoids in extracting solution, then from extracting solution, the molecular engram material being adsorbed with Ganoderma triterpenoids is extracted with ultrasonic extraction device
Material, after extraction, the Ganoderma triterpenoids of absorption on molecular engram material described in eluting；
Described molecular engram material comprises the following steps:
A, first dry Ganoderma is pulverized, is subsequently adding composite enzyme solution, carry out at 35～60 DEG C enzyme digestion reaction 1～
4h, afterwards heat inactivation compound enzyme；
The consumption of described REISH and the consumption of described compound enzyme, by described REISH 1g, the ratio of described compound enzyme 3mL
Calculate；
B, adding ethanol to step A gained, make concentration of alcohol reach 40～80%, regulation pH is 9～10, carry out supersound process 0.5～
Extraction fluid after 1h, described supersound process frequency is 20～40MHz, and temperature is 30～90 DEG C；
C, the process 1 of repetition step B～2 times；
D, being 2～5 by step C gained extracting solution regulation pH, cross resin and carry out adsorbing and eluting, eluent is concentrated into substance except alcohol
Long-pending 5～20% concentrated solution, add after the water of concentrated solution 3～4 times of volumes stirs and staticly settle, drying precipitate is obtained
Ganoderma triterpenoids；
E, take 1～1.5mmol Ganoderma triterpenoids and 6mmol methacrylic acid and be dissolved in 5～10ml chloroforms, be placed at 0 DEG C holding
30min, is subsequently adding 15～30mmol Ethylene glycol dimethacrylate and 15～30mmolN-N-isopropylacrylamides and 40
～100mg azodiisobutyronitrile, after mixing, it being passed through nitrogen, stirring is to being completely dissolved, and logical nitrogen 10min, sealing, at 65 DEG C of bars
Part reaction, until polymerization completely, is pulverized, is crossed 200 mesh sieves, is the methanol of 9:1 by volume ratio: alcohol mixeding liquid eluting Ganoderma triterpenoids,
I.e. can get molecular engram material.
In technique scheme, more specifically technical scheme is it may also is that described supersound extraction process control is at 250W,
40MHz。
Further: after extraction, first adsorb in institute with 50 DEG C of water elutions of 60 times of described molecular engram material weight
State the impurity on molecular engram material, more i.e. can get Ganoderma three with 25 DEG C of water elutions of 30 times of described molecular engram material weight
Terpene solution.
Owing to have employed technique scheme, the present invention compared with prior art has the advantages that
1, extraction Ganoderma triterpenoids purity and the extraction ratio of the present invention are the highest, and purity can reach 95～99%, extraction ratio can reach 85～
98%, operation simple and fast, imprinted material are reusable, are particularly suited for industrialized production.
2, the present invention extracts way technique environmental protection, does not use any organic reagent and prepares, on health without impact, same
Time imprinted material lowest critical solution temperature (LCST) be about 38 DEG C, supersound extraction because can accumulated heat, temperature is easy to reach
To more than 38 DEG C, without external heat source, energy-conserving and environment-protective.
Detailed description of the invention
Below in conjunction with specific embodiment, the invention will be further described:
The method that this molecular engram extracts Ganoderma triterpenoids with ultrasonic coupling first has to prepare molecular engram, and preparation method is as follows:
A, first the Ganoderma that 500g is dried is pulverized, be subsequently adding protease, cellulase and pectase with 4000:7:1 ratio
The compound enzyme 1.5L solution of example mixing, carries out enzyme digestion reaction 1h, afterwards heat inactivation compound enzyme at 35～60 DEG C；
B, adding ethanol to step A gained, make concentration of alcohol reach 80%, regulation pH is 9～10, carries out after supersound process 0.5h point
From extracting solution, described supersound process frequency is 40MHz, and temperature is 30 DEG C；
C, repeat the process 1 time of step B；
D, being 2～5 by step C gained extracting solution regulation pH, cross resin and carry out adsorbing and eluting, eluent is concentrated into substance except alcohol
Long-pending 20% concentrated solution, add after the water of 3 times of volumes of concentrated solution stirs and staticly settle, drying precipitate is obtained Ganoderma three
Terpene；
E, take 1mmol ganoderan and 6mmol methacrylic acid and be dissolved in 5ml chloroform, be placed at 0 DEG C holding 30min, then
Add 15mmol Ethylene glycol dimethacrylate and 15mmolN-N-isopropylacrylamide and 40mg azodiisobutyronitrile, mixing
After, it being passed through nitrogen, stirring, to being completely dissolved, is led to nitrogen 10min, sealing, at 65 DEG C of conditioned responses until polymerization is complete, is pulverized,
Cross 200 mesh sieves, be the methanol of 9:1 by volume ratio: alcohol mixeding liquid eluting Ganoderma triterpenoids, i.e. can get molecular engram material.
Then prepared molecular engram material and ultrasonic coupling are carried out the extraction of Ganoderma triterpenoids, extract process as follows:
Using REISH as extracting raw material, using water as extracting medium, utilize the molecular engram material adsorbing and extracting liquid prepared
In Ganoderma triterpenoids, then from extracting solution, extract the molecular engram material being adsorbed with Ganoderma triterpenoids with ultrasonic extraction device, extract
Process control is at 250W, 40MHz.After extraction, first with 50 DEG C of water elution absorption of 60 times of described molecular engram material weight
Impurity on described molecular engram material, more i.e. available clever with 25 DEG C of water elutions of 30 times of described molecular engram material weight
Sesame triterpene aqueous solution.
This molecular engram material is reusable.
Claims (3)
1. the method that a molecular engram extracts Ganoderma triterpenoids with ultrasonic coupling, it is characterised in that: using REISH as extraction
Raw material, using water as extracting medium, utilizes the Ganoderma triterpenoids in molecular engram material adsorbing and extracting liquid, then uses ultrasonic extraction device
The molecular engram material being adsorbed with Ganoderma triterpenoids is extracted, after extraction, on molecular engram material described in eluting from extracting solution
The Ganoderma triterpenoids of absorption；
Described molecular engram material comprises the following steps:
A, first dry Ganoderma is pulverized, is subsequently adding composite enzyme solution, carry out at 35～60 DEG C enzyme digestion reaction 1～
4h, afterwards heat inactivation compound enzyme；
The consumption of described REISH and the consumption of described compound enzyme, by described REISH 1g, the ratio of described compound enzyme 3mL
Calculate；
B, adding ethanol to step A gained, make concentration of alcohol reach 40～80%, regulation pH is 9～10, carry out supersound process 0.5～
Extraction fluid after 1h, described supersound process frequency is 20～40MHz, and temperature is 30～90 DEG C；
C, the process 1 of repetition step B～2 times；
D, being 2～5 by step C gained extracting solution regulation pH, cross resin and carry out adsorbing and eluting, eluent is concentrated into substance except alcohol
Long-pending 5～20% concentrated solution, add after the water of concentrated solution 3～4 times of volumes stirs and staticly settle, drying precipitate is obtained
Ganoderma triterpenoids；
E, take 1～1.5mmol Ganoderma triterpenoids and 6mmol methacrylic acid and be dissolved in 5～10ml chloroforms, be placed at 0 DEG C holding
30min, is subsequently adding 15～30mmol Ethylene glycol dimethacrylate and 15～30mmolN-N-isopropylacrylamides and 40
～100mg azodiisobutyronitrile, after mixing, it being passed through nitrogen, stirring is to being completely dissolved, and logical nitrogen 10min, sealing, at 65 DEG C of bars
Part reaction, until polymerization completely, is pulverized, is crossed 200 mesh sieves, is the methanol of 9:1 by volume ratio: alcohol mixeding liquid eluting Ganoderma triterpenoids,
I.e. can get molecular engram material.
The method that molecular engram the most according to claim 1 extracts Ganoderma triterpenoids with ultrasonic coupling, it is characterised in that: described
Supersound extraction process control is at 250W, 40MHz.
The method that molecular engram the most according to claim 1 extracts Ganoderma triterpenoids with ultrasonic coupling, it is characterised in that: extract
After, first with miscellaneous on described molecular engram material of 50 DEG C of water elutions absorption of 60 times of described molecular engram material weight
Matter, more i.e. can get Ganoderma triterpenoids solution with 25 DEG C of water elutions of 30 times of described molecular engram material weight.
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| CN201610500990.4A CN106074629A (en) | 2016-06-30 | 2016-06-30 | The method that molecular engram extracts Ganoderma triterpenoids with ultrasonic coupling |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106722875A (en) * | 2016-12-07 | 2017-05-31 | 盐城工学院 | The extracting method and Ganoderma lucidum of a kind of ganodenic acid |
| CN111574347A (en) * | 2020-05-27 | 2020-08-25 | 广西大学 | A molecular imprinting method for extracting jasmonone and jasmonolide from jasmine slag |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102772448A (en) * | 2012-08-15 | 2012-11-14 | 苏州菩芸生物科技有限公司 | Lucid ganoderma triterpene extraction method |
| CN103006717A (en) * | 2012-12-05 | 2013-04-03 | 深圳市岩代投资有限公司 | Method for extracting ingredients with lucid ganoderma effects |
| CN103664942A (en) * | 2013-12-16 | 2014-03-26 | 广西大学 | Method for extracting matrine by coupling molecular imprinting with ultrasonic extraction |
| CN105193867A (en) * | 2014-05-29 | 2015-12-30 | 南京中科药业有限公司 | Ergosterol-rich ganoderma lucidum spore oil and preparation method thereof |
-
2016
- 2016-06-30 CN CN201610500990.4A patent/CN106074629A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102772448A (en) * | 2012-08-15 | 2012-11-14 | 苏州菩芸生物科技有限公司 | Lucid ganoderma triterpene extraction method |
| CN103006717A (en) * | 2012-12-05 | 2013-04-03 | 深圳市岩代投资有限公司 | Method for extracting ingredients with lucid ganoderma effects |
| CN103664942A (en) * | 2013-12-16 | 2014-03-26 | 广西大学 | Method for extracting matrine by coupling molecular imprinting with ultrasonic extraction |
| CN105193867A (en) * | 2014-05-29 | 2015-12-30 | 南京中科药业有限公司 | Ergosterol-rich ganoderma lucidum spore oil and preparation method thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106722875A (en) * | 2016-12-07 | 2017-05-31 | 盐城工学院 | The extracting method and Ganoderma lucidum of a kind of ganodenic acid |
| CN111574347A (en) * | 2020-05-27 | 2020-08-25 | 广西大学 | A molecular imprinting method for extracting jasmonone and jasmonolide from jasmine slag |
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