CN105943568A - Acorn polyphenol extracting and purifying method - Google Patents
Acorn polyphenol extracting and purifying method Download PDFInfo
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- CN105943568A CN105943568A CN201610394852.2A CN201610394852A CN105943568A CN 105943568 A CN105943568 A CN 105943568A CN 201610394852 A CN201610394852 A CN 201610394852A CN 105943568 A CN105943568 A CN 105943568A
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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)
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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
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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Abstract
The invention discloses an acorn polyphenol extracting and purifying method. The method includes the steps that acorns are subjected to enzymolysis with cellulose and pectinase compound enzymes, an enzymolysis product is extracted with a microwave-assisted ethanol solution, after an obtained extracting solution is subjected to evaporation and concentration, water is added for dissolution, purification is conducted with an ultrafiltration membrane, and acorn polyphenol extracts are obtained. According to the method, the acorns serve as raw materials, the acorn polyphenol extracts are obtained efficiently through enzymolysis and microwave-assisted extraction in combination with ultrafiltration membrane purification and other technologies, the acorn polyphenol extracts with the polyphenol mass content being 35-45% can be obtained, and the method is easy to operate, low in cost, green and environmentally friendly and meets industrialized production requirements.
Description
Technical field
The present invention relates to the many phenol extractions of a kind of acorn nut and the method for purification, belong to natural plant composition and extract separation skill
Art field.
Background technology
Polyphenols is to find in plant food, and it has the compound promoting biological health benefit.Many
Aldehydes matter is present in the plant food that some are common, and plant polyphenol is for human body optimum health shape
The maintenance of state plays an important role.Polyphenols has a non-oxidizability, and inhibiting bacteria and diminishing inflammation and antiviral etc. are anti-ageing
Change and sun-proof physiologically active, there is good health-care effect.The huge function effect of polyphenols is drawn day by day
Play the great attention of scientist, thus cause the research and development enthusiasm to functional food.Functional food general
Read and incorporate diet all over the world very early, west have " make food become medicine, make medicament sources in
Food " viewpoint.The exploitation application of acorn nut polyphenol has important practical significance and wide market prospect.
Acorn nut is the general name referring to Fagaceae class seed.It is distributed widely in temperate zone and subtropical zone, mainly
In Asia.China is one of the widest country of the distribution area of acorn nut tree in the world, thus China's acorn nut resource is non-
The abundantest.Acorn nut nutrient constituents of fruit enriches, and research shows, in every 100g Fructus Querci acutissimae, about contains 8g albumen
Matter, 3g fat, 50.5g starch, 1.3g dietary fiber, 1.2g ash, 10.5g tannin, the most also contain
There are carotene, vitamin B1、B2With materials such as malic acids.Acorn nut is similar to rich in oils and fats, the oils squeezed out
Olive oil, is a kind of well edible oil.The biological function factor that acorn nut self is had, can resist, alleviate,
The heavy metal murders by poisoning to human body such as prevention lead, therefore the body and mind that edible acorn nut nourishing healthy based food can protect human body is good for
Health, improves the immunity of human body, improves intelligence and the developmental level of child.In acorn nut, the content of polyphenol substance reaches
To 2.2%～22%, it it is the raw material of a kind of preferable extraction polyphenols.In sum, from preparing acorn nut shallow lake
Extracting polyphenols in the filtrate produced during powder is a feasible and important job.
The method of the extraction of existing acorn nut polyphenols has a variety of, but is nearly all to use single extraction side
The shortcomings such as method, exists extraction efficiency low, purification difficult, operating time length.As: 1) hot water extraction method is very
Traditional method, the method have equipment be simple and convenient to operate, the advantage such as widely applicable, but there is also behaviour
Making the time long, the highest to the leaching rates of heterogeneity, repetitive process is many, inefficient, and energy consumption is high to be lacked
Point.2) organic solvent extraction needs consuming little energy, and product yield is high and purity is high, and its thick product easily divides
From purification.Extraction process is simple, invests little, it is adaptable to industrialized production, but increases organic solvent and be recycled into
This.3) yield of ultrasonic extraction is higher, and ultrasonic extraction is to utilize the character of ultrasound wave to strengthen to carry
Taking, ultrasonic extraction has that the time is short, efficiency high, and relatively hot water extraction method is easy.4) enzyme
It is that reaction condition is gentle that method extracts maximum advantage, and can carry out under non-organic solvent, and products therefrom is pure
Degree, stability, activity are the highest, pollution-free, but the enzyme extraction method cycle is longer, relatively inefficient.
Summary of the invention
The defect existed for prior art, it is an object of the invention to be to provide one to pass through with acorn nut for raw material
Enzymolysis, microwave radiation exaraction combine the method that the process efficient such as ultrafilter membrane purification obtain acorn nut polyphenol, and the method is grasped
Make simple, low cost, environmental protection, meet industrialization production requirements.
In order to realize above-mentioned technical purpose, the invention provides the many phenol extractions of a kind of acorn nut and the method for purification, should
Method is that acorn nut uses cellulase and pectase compound enzyme carry out enzymolysis, and enzymatic hydrolysate passes through microwave-assisted second
Alcoholic solution extracts, and gained extracting solution, after evaporation and concentration, is dissolved in water, and utilizes ultrafilter membrane to be purified, i.e.
Obtain acorn nut polyphenol extract.
Preferably method, the addition of cellulase and pectase compound enzyme is the 1～2% of acorn nut quality.
More preferably scheme, cellulase and pectase compound enzyme cellulase with the mass ratio of pectase are
(2～3): 1.
Further preferred scheme, hydrolysis temperature is 55～65 DEG C, and enzymolysis time is 30～40min.
Preferably scheme, microwave power is 200～400W.
Preferably scheme, ethanol solution mass percent concentration is 55～65%.
More preferably scheme, enzymatic hydrolysate with ethanol solution by solid-liquid ratio 1g:(20～30) mL mixes, micro-
Involve under 55～65 DEG C of temperature conditionss, after extracting 15～30min, filter, obtain filtrate I and filtering residue；Gained
Filtering residue extracts according to the extraction process of above-mentioned enzymatic hydrolysate, obtains filtrate II；Merging filtrate I and filtrate II,
It is evaporated concentrating.
Preferably scheme, use process that ultrafilter membrane is purified for be 25～35 DEG C in temperature, pressure be
Carry out under conditions of 0.1～0.2MPa.
More preferably scheme, is purified by retaining the ultrafilter membrane that molecule is 100kDa and realizes.
Further preferred scheme, acorn nut is through over cleaning, dry, pulverizing and screening and removing impurities pretreatment.
Preferably scheme, it is 0.30～0.5% that enriched product is diluted to mass percent concentration through being dissolved in water.
Hinge structure, the Advantageous Effects that technical scheme is brought:
1) technical scheme passes through cellulase and pectase complex enzyme zymohydrolysis, can be in a mild condition
Acorn nut epidermis cell structure is sufficiently destroyed, on this basis, in conjunction with microwave radiation exaraction, Polyphenols can be made
Compound can dissolution quick, efficient, substantially increase the extraction efficiency of polyphenols；And when shortening extraction
Between, solid-liquid ratio is little, and temperature is low, has met the theory of environmental protection.
2) technical scheme utilizes ultrafilter membrane to be purified extracting solution, it is possible at lower temperature and suitable
Realize the separation of polyphenol compound under the conditions of suitable pH, can effectively prevent polyphenol compound and other can give birth to
The degeneration of thing macromole, inactivation and self-dissolving, can obtain purity more much higher phenol material；And Ultra filtration membrane technology
There is advantage easy and simple to handle, with low cost, to be not required to increase any chemical reagent.
3) technical scheme, during extracting acorn nut polyphenol, can obtain acorn starch simultaneously
Deng product, acorn nut is made to be comprehensively utilized；Reduce the production cost of acorn nut polyphenol, reduce the wasting of resources, obtain
Obtain relatively high added value, increase economic efficiency.
4) technical scheme is with acorn nut for raw material by complex enzyme zymohydrolysis, microwave radiation exaraction, can obtain
Obtain polyphenol content extract more than 18.00%, further across ultrafilter membrane after purification, polyphenol can be obtained and contain
Amount extract more than 45%.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is further illustrated, but protection scope of the present invention is not limited to this.
Raw material of the present invention picks up from the Cyclobalanopsis robur acorn nut of hengyang, hunan province, raw material part nutritional labeling such as table
1:
The Cyclobalanopsis robur acorn nut part nutritional labeling of table 1 hengyang, hunan province
Embodiment 1:
1) pretreatment: filter out through being dried and the acorn nut of high-quality, clean, be dried, pulverize, screen out impurity
Standby.
2) cellulase of use sample quality 1% and the pectase of sample quality 0.5% are in 60 DEG C of constant temperature oscillations
Enzymolysis 35min in water-bath.In boiling water bath, 5min carries out enzyme denaturing process afterwards.Enzymolysis solution carries out next step
Reaction.
3) with 60% ethanol as solvent, be 1:25 at solid-liquid ratio, when microwave power is 300W, extract temperature
20min is extracted when degree is 60 DEG C.It is filtrated to get filtrate 1, filtering residue is repeated said process and obtains filtrate 2, close
And filtrate 1,2.
4) rotary evaporation concentrates: by 3) the rotated evaporation and concentration of extracting solution that obtains reclaims ethanol, obtains thick
Extract, adds water dissolving, obtains acorn nut Aqueous extracts.
5) ultrafilter membrane preliminary purification to acorn nut polyphenol: retaining molecule is that 100KDa film carries out acorn nut polyphenol
Ultrafiltration, by 4) in the feed concentration that obtains control to be 0.40%, temperature is 30 DEG C, and pressure is 0.141MPa.
Obtain acorn nut polyphenol substance after purification.
Survey polyphenol content by Prussia's blue laws and measure sample light absorption value under 540nm wavelength and gallic acid
Standard curve control, according to the concentration of total polyphenols in regression equation calculation sample solution, and calculates carrying of polyphenol
Take rate.In crude extract, the weight/mass percentage composition of polyphenol is 18.38%, the quality of polyphenol in extract after purification
Percentage composition is 45.12%.
Embodiment 2:
1) pretreatment: filter out through being dried and the acorn nut of high-quality, clean, be dried, pulverize, screen out impurity
Standby.
2) shake in 60 DEG C of constant temperature with the cellulase of sample quality 0.8% and the pectase of sample quality 0.3%
Swing enzymolysis 35min in water-bath.In boiling water bath, 5min carries out enzyme denaturing process afterwards.Enzymolysis solution carries out next step
Process.
3) with 60% ethanol as solvent, be 1:25 at solid-liquid ratio, when microwave power is 200W, extract temperature
20min is extracted when degree is 60 DEG C.It is filtrated to get filtrate 1, filtering residue is repeated said process and obtains filtrate 2, close
And filtrate 1,2.
4) rotary evaporation concentrates: by 3) the rotated evaporation and concentration of extracting solution that obtains reclaims ethanol, obtains thick
Extract, adds water dissolving, obtains acorn nut Aqueous extracts.
5) ultrafilter membrane preliminary purification to acorn nut polyphenol: retaining molecule is that 100KDa film carries out acorn nut polyphenol
Ultrafiltration, by 4) in the feed concentration that obtains control to be 0.50%, temperature is 25 DEG C, and pressure is 0.178MPa.
Obtain acorn nut polyphenol substance after purification.
Survey polyphenol content by Prussia's blue laws and measure sample light absorption value under 540nm wavelength and gallic acid
Standard curve control, according to the concentration of total polyphenols in regression equation calculation sample solution, and calculates carrying of polyphenol
Take rate.In crude extract, the weight/mass percentage composition of polyphenol is 8.2%, the quality hundred of polyphenol in extract after purification
Dividing content is 35.27%.
Embodiment 3:
1) pretreatment: filter out through being dried and the acorn nut of high-quality, clean, be dried, pulverize, screen out impurity
Standby.
2) cellulase of use sample quality 1% and the pectase of sample quality 0.5% are in 50 DEG C of constant temperature oscillations
Enzymolysis 35min in water-bath.In boiling water bath, 5min carries out enzyme denaturing process afterwards.
3) with 60% ethanol as solvent, be 1:30 at solid-liquid ratio, when microwave power is 300W, extract temperature
25min is extracted when degree is 60 DEG C.It is filtrated to get filtrate 1, filtering residue is repeated said process and obtains filtrate 2, close
And filtrate 1,2.
4) rotary evaporation concentrates: by 3) the rotated evaporation and concentration of extracting solution that obtains reclaims ethanol, obtains thick
Extract, adds water dissolving, obtains acorn nut Aqueous extracts.
5) ultrafilter membrane preliminary purification to acorn nut polyphenol: retaining molecule is that 100KDa film carries out acorn nut polyphenol
Ultrafiltration, by 4) in the feed concentration that obtains control to be 0.30%, temperature is 25 DEG C, and pressure is 0.108MPa.
Obtain acorn nut polyphenol substance after purification.
Survey polyphenol content by Prussia's blue laws and measure sample light absorption value under 540nm wavelength and gallic acid
Standard curve control, according to the concentration of total polyphenols in regression equation calculation sample solution, and calculates carrying of polyphenol
Take rate.In crude extract, the weight/mass percentage composition of polyphenol is 14.65%, the quality of polyphenol in extract after purification
Percentage composition is 39.41%.
Claims (10)
1. the many phenol extractions of acorn nut and the method for purification, it is characterised in that: acorn nut is used cellulase and pectin
Enzyme compound enzyme carries out enzymolysis, and enzymatic hydrolysate is extracted by Microwave-Assisted Ethanol solution, and gained extracting solution is through pervaporation
After concentration, it is dissolved in water, uses ultrafilter membrane to be purified, obtain acorn nut polyphenol extract.
The many phenol extractions of acorn nut the most according to claim 1 and the method for purification, it is characterised in that: described fibre
Dimension element enzyme and addition is acorn nut quality the 1～2% of pectase compound enzyme.
The many phenol extractions of acorn nut the most according to claim 2 and the method for purification, it is characterised in that: described fibre
The mass ratio of dimension element enzyme and pectase compound enzyme cellulase and pectase is (2～3): 1.
4. according to the many phenol extractions of acorn nut described in any one of claims 1 to 3 and the method for purification, it is characterised in that:
Described hydrolysis temperature is 55～65 DEG C, and enzymolysis time is 30～40min.
The many phenol extractions of acorn nut the most according to claim 1 and the method for purification, it is characterised in that: described is micro-
Wave power is 200～400W.
The many phenol extractions of acorn nut the most according to claim 1 and the method for purification, it is characterised in that: described second
Alcoholic solution mass percent concentration is 55～65%.
7. according to the many phenol extractions of acorn nut described in claim 1,5 or 6 and the method for purification, it is characterised in that: enzyme
Hydrolysis products with ethanol solution by solid-liquid ratio 1g:(20～30) mL mixes, in microwave and 55～65 DEG C of temperature conditionss
Under, after extracting 15～30min, filter, obtain filtrate I and filtering residue；Gained filtering residue is according to above-mentioned enzymatic hydrolysate
Extraction process extract, obtain filtrate II；Merging filtrate I and filtrate II, be evaporated concentrating.
The many phenol extractions of acorn nut the most according to claim 1 and the method for purification, it is characterised in that: use ultrafiltration
The process that film is purified temperature be 25～35 DEG C, pressure be to carry out under conditions of 0.1～0.2MPa.
9. according to the many phenol extractions of the acorn nut described in claim 1 or 8 and the method for purification, it is characterised in that: described
Be purified by retain ultrafilter membrane that molecule is 100kDa and realize.
10. according to the many phenol extractions of a kind of acorn nut described in claims 1 to 3,5,6,8 any one and purification process thereof,
It is characterized in that: described acorn nut through over cleaning, be dried, pulverize and screen out impurity pretreatment.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107149150A (en) * | 2017-05-22 | 2017-09-12 | 辽宁大学 | Anti-lung cancer health products containing rice bran polyphenol |
| CN107158161A (en) * | 2017-05-22 | 2017-09-15 | 辽宁大学 | Anti-lung cancer health products containing rice husk polyphenol |
| CN111944165A (en) * | 2020-07-15 | 2020-11-17 | 汕头大学 | Polyphenol tyrosinase inhibitor and extraction method and application thereof |
| CN112250731A (en) * | 2020-10-22 | 2021-01-22 | 贵州省生物技术研究所(贵州省生物技术重点实验室、贵州省马铃薯研究所、贵州省食品加工研究所) | Green functional factor food preparation equipment and preparation method thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107149150A (en) * | 2017-05-22 | 2017-09-12 | 辽宁大学 | Anti-lung cancer health products containing rice bran polyphenol |
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| CN112250731A (en) * | 2020-10-22 | 2021-01-22 | 贵州省生物技术研究所(贵州省生物技术重点实验室、贵州省马铃薯研究所、贵州省食品加工研究所) | Green functional factor food preparation equipment and preparation method thereof |
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Application publication date: 20160921 |