CN107519232A - One kind extraction Gueldenstaedtia verna extractive of general flavone and preparation method thereof - Google Patents
One kind extraction Gueldenstaedtia verna extractive of general flavone and preparation method thereof Download PDFInfo
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- CN107519232A CN107519232A CN201710959263.9A CN201710959263A CN107519232A CN 107519232 A CN107519232 A CN 107519232A CN 201710959263 A CN201710959263 A CN 201710959263A CN 107519232 A CN107519232 A CN 107519232A
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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)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
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- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/17—Preparation or pretreatment of starting material involving drying, e.g. sun-drying or wilting
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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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- 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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- 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 present invention relates to a kind of Gueldenstaedtia verna extractive of general flavone and its extracting method, belong to technical field of traditional Chinese medicines；This method mainly includes the following steps that：(1) pre-treatment of Gueldenstaedtia verna medicinal material；(2) assisted microwave synthesis method extraction Gueldenstaedtia verna general flavone is digested；(3) macroporous absorbent resin elutes, and is dried in vacuo and produces Gueldenstaedtia verna extractive of general flavone.The present invention carries out enzymolysis pretreatment to Gueldenstaedtia verna using cellulose and pectin compound enzyme before Microwave Extraction, Gueldenstaedtia verna after processing carries out Microwave Extraction and macroporous resin purification again, for Gueldenstaedtia verna general flavone content after extraction purification up to more than 60%, recovery rate obtains larger raising.Present invention enzymolysis, extraction, purification process are simple to operate, and production cost reduces, and is easy to be industrially used and promotes.
Description
Technical field
The invention belongs to Separation of Natural Products field, is related to a kind of extraction Gueldenstaedtia verna extractive of general flavone and its preparation side
Method.
Background technology
Gueldenstaedtia verna is legume Gueldenstaedtia verna Gueldenstaedtia verna (Georgi) A.Bor. drying herb,
Summer, the excavation of season in autumn two.Begin to carry《Compendium of Materia Medica》.Originate in the ground such as northeast, North China, East China, the Shaanxi middle and south, In Eastern Gansu.Rice mouth
Bag is sweet, bitter, cold in nature, has the effect of clearing heat and detoxicating, cures mainly carbuncle swells furunculosis, external auditory meatus furuncle, appendicitis.
The research of Gueldenstaedtia verna chemical composition shows that it contains the various actives such as flavones terpene, volatile oil, sterol and organic acid
Composition, wherein flavones ingredient are its main pharmacodynamics compositions.Pharmacological research show Gueldenstaedtia verna have antibacterial, it is anti-oxidant, antiviral,
Analgesia, anti-inflammatory isoreactivity.
In the prior art, Chinese patent 201621290486.8 discloses a kind of Gueldenstaedtia verna cycling extraction device, using more
The technology of secondary circulation distillation extraction, Gueldenstaedtia verna is carried out the soak extraction of multiple fresh extractor liquid, and extraction effect is good, takes short.
Shi Meirong optimizes using general flavone in ultrasonic assistant method extraction Gueldenstaedtia verna and to technique.On the basis of experiment of single factor
On, determine solid-liquid ratio, ultrasonic temperature, ultrasonic power, 4 factors of ultrasonic time Box~benhnken experimental design.With total
The recovery rate of flavones is response, optimizes the extraction process of Gueldenstaedtia verna general flavone using response phase method, establishes and analyze each factor
With the mathematical modeling of desired value.Determine that optimal processing parameter is：Solid-liquid ratio 1:40g/mL, 45 DEG C of ultrasonic temperature, ultrasonic power
300W, ultrasonic time 60min, general flavone recovery rate theoretical value 8.08%, actual value 8.22%, relative standard deviation are
0.33% (ultrasonic extraction process of the U.S. flourish response phase methods optimization Gueldenstaedtia verna general flavone of history, Chinese agronomy circular, 2017,33
(23):129~137.).
Therefore, in order to preferably develop and utilize Gueldenstaedtia verna, and material base is provided for the further exploitation of its Related product
Plinth, pharmacological action is given full play to, the research to Gueldenstaedtia verna total flavone extracting method is always that those skilled in the art are worth probing into
The problem of.The present invention is that Gueldenstaedtia verna general flavone is extracted by digesting assisted microwave synthesis method, then it is entered by macroreticular resin
Row purifying, this method is simple, feasible, to promote the exploitation of Gueldenstaedtia verna medical value to have positive realistic meaning.
The content of the invention
It is an object of the invention to provide a kind of Gueldenstaedtia verna extractive of general flavone and its extracting method.
The present invention is achieved by the following technical solutions：
A kind of Gueldenstaedtia verna extractive of general flavone and its extracting method, comprise the following steps：
(4) by Gueldenstaedtia verna drying, crushing, complex enzyme and buffer solution is added, carries out enzymolysis pretreatment, is filtered, into filter residue
Add ethanol, Microwave Extraction, filtering, merging filtrate；
(5) filtrate decompression concentrates, and macroporous resin column absorption, is first eluted with water resin column, then use ethanol elution on concentrate
Resin column, collect eluent；
(6) eluent concentrates, and vacuum drying obtains Gueldenstaedtia verna general flavone.
Complex enzyme addition in the step (1) is the 1.3%~2.2% of quality of medicinal material.
The complex enzyme is cellulase:Pectase is 1:1~3:1.
Buffer solution in the step (1) is Acetic acid-sodium acetate solution, and pH value is 4.0~4.6.
Enzymolysis in the step (1), hydrolysis temperature are 40~60 DEG C, 60~80min of enzymolysis time.
Concentration of alcohol is 60~80% (v/v) in the step (1).
Microwave Extraction in the step (1), microwave power are 380~420W；The Microwave Extraction time is 4~7min.
Macroreticular resin is D101 macroreticular resins in the step (2).
A kind of Gueldenstaedtia verna extractive of general flavone and its extracting method, comprise the following steps：
(1) take Gueldenstaedtia verna medicinal material drying, be ground into 40~60 mesh, obtain Gueldenstaedtia verna powder.Gueldenstaedtia verna powder is weighed, according to
Quality of medicinal material is than adding 1.3~2.2% complex enzyme (cellulases:Pectase=1:1~3:1) 8~12 times of amounts, pH value, are added
For 4~4.6 Acetic acid-sodium acetate buffer solution, 60~80min of Gueldenstaedtia verna medicinal material is digested in the case where hydrolysis temperature is 40~60 DEG C of regulations
Afterwards, filter, 15~25 times of 60~80% ethanol of amount are added into filter residue, microwave carries under the conditions of microwave power is 380~420W
4~7min is taken, is filtered, merging filtrate, filtrate decompression concentration.
(2) extract solution is concentrated into 0.5~1.5g/ml, from D101 macroreticular resins, loading volume and macroreticular resin quality
Ratio is 8:1~12：1, wet method loading, loading adsorbs 6~8h, is first washed with 4~8BV distilled water, washing speed is 3~5BV/
H, it is then 40~70% ethanol elutions with 3~5BV volume fractions, elution speed is 3~5BV/h.
(3) by the ethanol eluate of collection, concentrate, vacuum drying, you can obtain Gueldenstaedtia verna extractive of general flavone.
Preferably, a kind of extracting method of Gueldenstaedtia verna general flavone, specifically includes following steps：
(1) by Gueldenstaedtia verna medicinal material drying, be ground into 40~60 mesh, obtain Gueldenstaedtia verna powder.According to quality of medicinal material than adding
0.78% complex enzyme (cellulase:Pectase=2:1) 10 times of amounts, are added, the Acetic acid-sodium acetate buffer solution that pH value is 4.3,
Hydrolysis temperature is under 49 DEG C of regulations, after digesting Gueldenstaedtia verna medicinal material 68min, filtering, 20 times of 70% ethanol of amount is added into filter residue,
Microwave power is Microwave Extraction 5min under the conditions of 400W, is filtered, merging filtrate.
(2) filtrate decompression is concentrated, and extract solution is concentrated into 1.0g/ml, from D101 macroreticular resins, loading volume and macropore
Resin quality ratio is 10：1 wet method loading, loading absorption 7h, is first washed, washing speed is 4BV/h, then with 6BV distilled water
With 4BV 60% (v/v) ethanol elution, elution speed 4BV/h, eluent is collected；
(3) by the ethanol eluate of collection, concentrate, vacuum drying, you can obtain Gueldenstaedtia verna general flavone product.
Beneficial effects of the present invention：
(1) present invention is using cellulose and pectin compound enzyme, and a kind of product of enzyme can in this multienzyme complex system
With the substrate as another enzyme, so that enzymolysis is more successfully carried out.Cellulase is improving cellulose, hemicellulose
While element decomposes, can promote the dissolving of plant cell wall makes more plant cell Dissolve things insides be dissolved out and will can be not easy to disappear
The macromolecular polysaccharide of change, protein and lipid degradation are into small-molecule substance；And hemicellulose and pectin can produce after being partially soluble in water
Raw viscosity solution, addition pectase can cooperate with cellulase to reduce viscosity, increase the diffusion of endogenous enzymes, improve enzymolysis efficiency.
(2) present invention utilizes bio-enzyme degradation principle, and enzymolysis pretreatment is carried out before Microwave Extraction to Gueldenstaedtia verna, through processing
After Gueldenstaedtia verna afterwards carries out Microwave Extraction again, the recovery rate of Gueldenstaedtia verna general flavone obtains larger raising.
(3) present invention combines macroporous resin purification method, and the Gueldenstaedtia verna general flavone content of extraction purification is up to more than 60%；This hair
Bright extraction, purification process are simple to operate, and production cost reduces, and is easy to be industrially used and promotes.
Embodiment
Come with reference to embodiment to further detailed description of the present invention, but claimed scope is not only limited to institute
The description of work.
Embodiment 1
Gueldenstaedtia verna general flavone extraction step is as follows：
(1) take Gueldenstaedtia verna medicinal material drying, be ground into 50 mesh, obtain Gueldenstaedtia verna powder.Gueldenstaedtia verna powder 50g is weighed, according to
Quality of medicinal material is than adding 0.78% complex enzyme (cellulase:Pectase=2:1) acetic acid that, 10 times of addition is measured, pH value is 4.3-
Sodium-acetate buffer, after Gueldenstaedtia verna medicinal material 68min is digested under hydrolysis temperature is 49 DEG C of regulations, filtering, 20 are added into filter residue
70% ethanol of amount again, the Microwave Extraction 5min under the conditions of microwave power is 400W, filtering, merging filtrate, filtrate decompression concentration.
(2) extract solution is concentrated into 1.0g/ml, from D101 macroreticular resins, loading volume and macroreticular resin mass values
For 10：1 wet method loading, loading absorption 7h, is first washed, washing speed is 4BV/h, then with 4BV volume fractions with 6BV distilled water
For 60% ethanol elution, elution speed 4BV/h.
(3) by the ethanol eluate of collection, concentrate, vacuum drying, you can obtain the product of Gueldenstaedtia verna general flavone purity 72%.
Embodiment 2
Gueldenstaedtia verna general flavone extraction step is as follows：
(1) take Gueldenstaedtia verna medicinal material drying, be ground into 60 mesh, obtain Gueldenstaedtia verna powder.Gueldenstaedtia verna powder 50g is weighed, according to
Quality of medicinal material is than adding 2.2% complex enzyme (cellulase:Pectase=3:1) 12 times of amounts, are added, acetic acid-vinegar that pH value is 4.6
Sour sodium buffer solution, after Gueldenstaedtia verna medicinal material 80min is digested under hydrolysis temperature is 60 DEG C of regulations, filtering, 25 times are added into filter residue
80% ethanol is measured, the Microwave Extraction 7min under the conditions of microwave power is 420W, filtering, merging filtrate, filtrate decompression concentration.
(2) extract solution is concentrated into 1.5g/ml, from D101 macroreticular resins, loading volume and macroreticular resin mass values
For 12：1 wet method loading, loading absorption 8h, is first washed, washing speed is 5BV/h, then with 5BV volume fractions with 8BV distilled water
For 70% ethanol elution, elution speed 5BV/h.
(3) by the ethanol eluate of collection, concentrate, vacuum drying, you can obtain the product of Gueldenstaedtia verna general flavone purity 70%.
Embodiment 3
Gueldenstaedtia verna general flavone extraction step is as follows：
(1) take Gueldenstaedtia verna medicinal material drying, be ground into 40 mesh, obtain Gueldenstaedtia verna powder.Gueldenstaedtia verna powder 50g is weighed, according to
Quality of medicinal material is than adding 1.3% complex enzyme (cellulase:Pectase=1:1) 8 times of amounts, are added, acetic acid-acetic acid that pH value is 4
Sodium buffer solution, after Gueldenstaedtia verna medicinal material 60min is digested under hydrolysis temperature is 40 DEG C of regulations, filtering, 15 times of amounts are added into filter residue
60% ethanol, the Microwave Extraction 4min under the conditions of microwave power is 380W, filtering, merging filtrate, filtrate decompression concentration.
(2) extract solution is concentrated into 0.5g/ml, from D101 macroreticular resins, loading volume and macroreticular resin mass values
For 8：1 wet method loading, loading absorption 6h, is first washed, washing speed is 3BV/h, then with 3BV volume fractions with 4BV distilled water
For 40% ethanol elution, elution speed 3BV/h.
(3) by the ethanol eluate of collection, concentrate, vacuum drying, you can obtain the product of Gueldenstaedtia verna general flavone purity 67%.
Embodiment 4
Gueldenstaedtia verna general flavone extraction step is as follows：
(1) take Gueldenstaedtia verna medicinal material drying, be ground into 60 mesh, obtain Gueldenstaedtia verna powder.Gueldenstaedtia verna powder 50g is weighed, according to
Quality of medicinal material is than adding 1.3% complex enzyme (cellulase:Pectase=3:1) 8 times of amounts, are added, acetic acid-vinegar that pH value is 4.6
Sour sodium buffer solution, after Gueldenstaedtia verna medicinal material 80min is digested under hydrolysis temperature is 40 DEG C of regulations, filtering, 15 times are added into filter residue
80% ethanol is measured, the Microwave Extraction 7min under the conditions of microwave power is 380W, filtering, merging filtrate, filtrate decompression concentration.
(2) extract solution is concentrated into 0.5g/ml, from D101 macroreticular resins, loading volume and macroreticular resin mass values
For 12：1 wet method loading, loading absorption 6h, is first washed, washing speed is 3BV/h, then with 5BV volume fractions with 8BV distilled water
For 40% ethanol elution, elution speed 5BV/h.
(3) by the ethanol eluate of collection, concentrate, vacuum drying, you can obtain the product of Gueldenstaedtia verna general flavone purity 68%.
Embodiment 5
Gueldenstaedtia verna general flavone extraction step is as follows：
(1) take Gueldenstaedtia verna medicinal material drying, be ground into 40 mesh, obtain Gueldenstaedtia verna powder.Gueldenstaedtia verna powder 50g is weighed, according to
Quality of medicinal material is than adding 2.2% complex enzyme (cellulase:Pectase=1:1) 12 times of amounts, are added, acetic acid-acetic acid that pH value is 4
Sodium buffer solution, after Gueldenstaedtia verna medicinal material 60min is digested under hydrolysis temperature is 60 DEG C of regulations, filtering, 25 times of amounts are added into filter residue
60% ethanol, the Microwave Extraction 4min under the conditions of microwave power is 420W, filtering, merging filtrate, filtrate decompression concentration.
(2) extract solution is concentrated into 1.5g/ml, from D101 macroreticular resins, loading volume and macroreticular resin mass values
For 8：1 wet method loading, loading absorption 8h, is first washed, washing speed is 5BV/h, then with 3BV volume fractions with 4BV distilled water
For 70% ethanol elution, elution speed 3BV/h.
(3) by the ethanol eluate of collection, concentrate, vacuum drying, you can obtain the product of Gueldenstaedtia verna general flavone purity 70%.
Claims (10)
1. a kind of extracting method of Gueldenstaedtia verna general flavone, it is characterised in that comprise the following steps：
(1) by Gueldenstaedtia verna drying, crushing, complex enzyme and buffer solution is added, carries out enzymolysis pretreatment, filters, is added into filter residue
Ethanol, Microwave Extraction, filtering, merging filtrate；
(2) filtrate decompression concentrates, macroporous resin column absorption on concentrate, is first eluted with water resin column, then with ethanol elution resin
Post, collect eluent；
(3) eluent concentrates, and vacuum drying obtains Gueldenstaedtia verna general flavone.
2. extracting method as claimed in claim 1, it is characterised in that the complex enzyme addition in the step (1) is medicinal material
The 1.3%~2.2% of quality.
3. extracting method as claimed in claim 2, it is characterised in that the complex enzyme is the combination of cellulase and pectase
Thing.
4. extracting method as claimed in claim 3, it is characterised in that the cellulase:Pectase is 1:1~3:1.
5. extracting method as claimed in claim 1, it is characterised in that the buffer solution in the step (1) is Acetic acid-sodium acetate
Solution, pH value are 4.0~4.6.
6. extracting method as claimed in claim 1, it is characterised in that the enzymolysis in the step (1), hydrolysis temperature be 40~
60 DEG C, 60~80min of enzymolysis time.
7. extracting method as claimed in claim 1, it is characterised in that concentration of alcohol is 60~80% (v/ in the step (1)
v)。
8. extracting method as claimed in claim 1, it is characterised in that Microwave Extraction in the step (1), microwave power are
380~420W；The Microwave Extraction time is 4~7min.
9. extracting method as claimed in claim 1, it is characterised in that macroreticular resin is D101 macropore trees in the step (2)
Fat.
10. extracting method as claimed in claim 1, it is characterised in that comprise the following steps：
(1) take Gueldenstaedtia verna medicinal material drying, be ground into 40~60 mesh, obtain Gueldenstaedtia verna powder.Gueldenstaedtia verna powder is weighed, according to medicinal material
Mass ratio adds 1.3~2.2% complex enzyme (cellulases:Pectase=1:1~3:1), add 8~12 times amount, pH value be 4~
4.6 Acetic acid-sodium acetate buffer solution, after 60~80min of Gueldenstaedtia verna medicinal material is digested under hydrolysis temperature is 40~60 DEG C of regulations, mistake
Filter, into filter residue add 15~25 times amount 60~80% ethanol, microwave power be 380~420W under the conditions of Microwave Extraction 4~
7min, filtering, merging filtrate, filtrate decompression concentration；
(2) extract solution is concentrated into 0.5~1.5g/ml, from D101 macroreticular resins, loading volume and macroreticular resin mass values
For 8:1~12：1, wet method loading, loading adsorbs 6~8h, is first washed with 4~8BV distilled water, washing speed is 3~5BV/h, so
It is afterwards 40~70% ethanol elutions with 3~5BV volume fractions, elution speed is 3~5BV/h；
(3) by the ethanol eluate of collection, concentrate, vacuum drying, you can obtain Gueldenstaedtia verna extractive of general flavone.
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| CN111419892A (en) * | 2020-03-31 | 2020-07-17 | 右江民族医学院 | Extraction method of abrus herb total flavone |
| WO2022227286A1 (en) * | 2021-04-27 | 2022-11-03 | 云南英格生物技术有限公司 | Preparation method for pyracantha fruit extract, and use |
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