CN105949163B - The method for extraction and purification of anthocyanidin in a kind of Black Box Tracing pomace - Google Patents
The method for extraction and purification of anthocyanidin in a kind of Black Box Tracing pomace Download PDFInfo
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- CN105949163B CN105949163B CN201610342698.4A CN201610342698A CN105949163B CN 105949163 B CN105949163 B CN 105949163B CN 201610342698 A CN201610342698 A CN 201610342698A CN 105949163 B CN105949163 B CN 105949163B
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
- C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
- C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B61/00—Dyes of natural origin prepared from natural sources, e.g. vegetable sources
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- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention relates to a kind of new method of anthocyanidin extraction purification in Black Box Tracing pomace, ultramicro grinding, enzymolysis and extraction, low-speed centrifugal, high speed centrifugation, ultrafiltration impurity elimination, macroporous resin purification, nanofiltration, freeze-drying are specifically included.Pectase and cellulase degradation is added more than after 300 mesh in the average particle size that pomace is crushed to material using vibration type ultramicro disintegrator.Obtained enzymolysis solution is successively using separation decanter centrifuge and self-cleaning type disk centrifuge, low speed and high speed centrifugation.Clear solution ultrafiltration.The efflux of ultrafiltration macroreticular resin chromatographic column adsorbs, with 60%~90% ethanol elution anthocyanidin.Obtained anthocyanidin ethanol solution NF membrane nanofiltration removes etoh solvent.Anthocyanidin solution lyophilization or spray drying process dry trapped fluid, until moisture content of finished products content≤5%.The Black Box Tracing anthocyanidin powder of aubergine is obtained, the wherein content of anthocyanidin is 60% 90%.
Description
Technical field
The present invention relates to a kind of method for extraction and purification of anthocyanidin in Black Box Tracing pomace, belong to processing of farm products skill
Art field.
Background technology
The Black Box Tracing category rose family, is perennial machaka, and fruit is atropurpureus berry.The plant originates in
There are plantation in Northeastern United States, the provinces and cities such as present China Jilin, Heilungkiang, Liaoning, Inner Mongol, Gansu, Jiangsu.The seeds adaptability
By force, growth is rapid, tree-like beauty, resists cold drought-enduring, is suitable as ornamental trees.In addition, its fruit is rich in multivitamin, amino
The a variety of nutrition of acid, minerals, organic acid, terpene, carrotene and functional component.It is noted that the fruit also contains polyphenol
Substance is up to 2.5% or more, and wherein pattern cellulose content is of about 1%.In recent years, studies have shown that the fruit has improvement eyesight and sleeps
It sleeps, alleviates senile dementia, the diseases effects such as treatment heart disease, hypertension and cancer.Especially, which is rich in more with it
Aldehydes matter especially anthocyanidin is related, and the bioactivity of the anthocyanidin is higher than the anthocyanidin extracted in the plants such as purple sweet potato.Mesh
Before, it is widely used in food, medicine and cosmetics in quickbeam and its extract.
China from successful foreign introduces Black Box Tracing seedling, by the Technique Popularizing in more than ten years, big face at present
Product plantation.The trees serve not only as a kind of ornamental trees, but also as a kind of industrial crops.Some Enterprises are opened using its fruit
The deep processed products such as Sorbus alnifloria fruit juice, fruit wine and fruit vinegar are sent out, good economic benefit is obtained, and meet diversified market need
It asks.However, at present after these fruit deep processings, pomace is as just feed or fertilizer.The pomace is attached not by deep processing
It is value added limited.It is noted that containing abundant anthocyanidin in the pomace.Therefore, anthocyanidin is extracted using Sorbus alnifloria pomace, no
The economic benefit for only improving associated production enterprise, improves the added value of pomace, and meet domestic and international market to high bioactivity
Sorbus alnifloria anthocyanidin Related product demand.
Within past more than 30 year, the mankind successfully extract and are purified into a variety of anthocyanidin from plant.To mesh
Before until, these method generally existing environmental pollutions, extraction purification efficiency is low and yield is low, method is loaded down with trivial details, solvent can not recycle
The shortcomings of, it is difficult to be applied in actual production.Especially, impurity is not efficiently removed in purification process, causes to spend
The purity of green element is low, and the price of serious limit product influences its application in the related art.
Cerise or aubergine is presented in acid condition in anthocyanidin or a kind of safe water-soluble natural pigment.
The harmful synthetic food colors such as alternative existing amaranth, famille rose.Therefore, largely anthocyanidin is developed especially
Sorbus alnifloria anthocyanidin, can not only meet people to it as drug, health food and cosmetics the needs of, but also it is alternative to artificial
The use of synthetic dyestuff will bring glad tidings to the improvement of the health of the mankind.
Invention content
It is an object of the present invention to provide a kind of method for extraction and purification of anthocyanidin in Black Box Tracing pomace.
In order to achieve the above objectives, the technical solution adopted by the present invention is：Anthocyanidin in a kind of Black Box Tracing pomace
Method for extraction and purification includes the following steps：
（1）Black Box Tracing is beaten to obtain pulp；
（2）Fruit juice in the pulp is detached with pomace；
（3）The pomace is added in vibration type ultramicro disintegrator, it is going for 2~10 times of the pomace then to add volume
After ionized water, the average particle size of ultramicro grinding 30-40 minutes to material is greater than or equal to 300 mesh；
（4）Pomace after ultramicro grinding is added in enzymatic vessel, with the pH value of citric acid regulating solution to 2.5~4.5；It connects
Get off under the conditions of temperature is 25~55 DEG C, pectase and concentration of polymer solution that concentration of polymer solution is 0.01~0.3% is added
It for 0.01~0.2% cellulase, is uniformly mixed, until there is no pectin to be detected in solution, obtains enzymolysis solution；
（5）The enzymolysis solution is centrifuged under 3000-5000r/min rotating speeds, obtains thick clear solution；
（6）The thick clear solution is centrifuged under 5000-13000r/min rotating speeds, obtains clear solution；
（7）The clear solution is added in the ultrafiltration apparatus that molecular cut off is 3-10kDa and carries out ultrafiltration, removal remaining
Impurity；Control pressure is 0.1~0.7MPa, and temperature is 10~50 DEG C, until without anthocyanidin outflow；
（8）By step（7）Efflux styrene type non-polar macroporous resin or styrene type low pole macropore is added
It in resin chromatography column, is first eluted, is removed not by the impurity of macroporous resin adsorption with pure water or deionized water；Use volume dense again
The ethanol elution that degree is 60%~90% so that anthocyanidin is eluted, and eluent is anthocyanidin ethanol solution；
（9）The NF membrane that molecular cut off is 200-300Da is added in anthocyanidin ethanol solution, carries out nanofiltration, removal is molten
Agent ethyl alcohol, operating pressure is 0.1~0.7MPa, temperature is 10~50 DEG C；When anthocyanidin volumes of aqueous ethanol is reduced to original body
Long-pending 1/10th stop nanofiltration and operate to obtain anthocyanidin solution；
（10）Anthocyanidin solution is dried up to product using vacuum freeze-drying method or spray drying process.
Preferably technical solution is：The Black Box Tracing be fresh Black Box Tracing or freezing after solve
The Black Box Tracing of jelly.
Preferably technical solution is：The fruit juice in the pulp is divided with pomace using slurry slag separator or centrifuge
From.
Preferably technical solution is：In step（7）In, when still there is anthocyanidin outflow, adds deionized water and continue to surpass
Filter.
Preferably technical solution is：The vacuum freeze-drying method refers to 0 DEG C by anthocyanidin solution precooling hereinafter, so
It is less than or equal to 50kPa in vacuum degree afterwards, under the conditions of -20~-40 DEG C, freeze-drying to moisture content of finished products content≤5%.
Preferably technical solution is：The spray drying process refers to anthocyanidin solution being sprayed dry in 110 ~ 120 DEG C of conditions
It is dry to moisture content of finished products content≤5%.The Black Box Tracing anthocyanidin powder of aubergine is obtained, the wherein content of anthocyanidin is 60-
90%。
Since above-mentioned technical proposal is used, the present invention has following advantages and effect compared with prior art：
1, the present invention is fully crushed Sorbus alnifloria pomace using superfine communication technique, effectively realizes the imperceptibility of pomace；Add
Add pectase and cellulase hydrolysis solution, effectively realizes the release of contained anthocyanidin in pectin macromolecular colloid；It uses successively
Low-speed centrifugal, high speed centrifugation and ultrafiltration filtering, impurity contained in solution is largely removed efficiently；Pass through macropore tree
Fat adsorbs anthocyanidin, and abjection is washed with water in other water-solubility impurities, ensures that product has higher purity（More than or equal to 60）.
2, the invention enables anthocyanidin resources in Black Box Tracing pomace to be utilized effectively.The present invention, which adopts, to be extracted with water
Method can efficiently extract the anthocyanidin in Black Box Tracing pomace, accomplish scale production.Black Box Tracing anthocyanidin can
It is directly edible, it also can be used as auxiliary material or natural pigment be added in beverage, FD ＆ C, improve black fruit gland rib
The added value of Sorbus alnifloria pomace.
Specific implementation mode
With reference to embodiment, the invention will be further described：
Embodiment：The method for extraction and purification of anthocyanidin in a kind of Black Box Tracing pomace
（1）The Black Box Tracing mashing that freezing is thawed, obtains pulp.
（2）Pulp is detached fruit juice with black slag using slurry slag separator or centrifuge.
（3）Pomace is added in vibration type ultramicro disintegrator, after the deionized water for adding 2 times of volumes, ultramicro grinding 30 is divided
Clock, until the average particle size of material is more than 300 mesh.
（4）Pomace after ultramicro grinding is added in enzymatic vessel, with the pH to 2.5 of citric acid regulating solution；25 DEG C of temperature
Under the conditions of, the pectase and 0.01% cellulase of solution quality 0.01% is added, is uniformly mixed, digests 2 hours, until not having in solution
Until thering is pectin to be detected, enzymolysis solution is obtained.
（5）Enzymolysis solution is used into separation decanter centrifuge, is centrifuged under 3000r/min rotating speeds, removal is most of
Residue obtains thick clear solution.
（6）Clear solution is used into self-cleaning type disk centrifuge, is centrifuged under 5000r/min rotating speeds, removal is remaining
A small amount of residue, obtain clear solution.
（7）Clear solution is added in the ultrafiltration apparatus that molecular cut off is 3kDa, carries out ultrafiltration, further removal remaining
The big molecular impurities such as a small amount of polysaccharide, protein, cellulose, pectin.Control pressure is 0.1MPa, and temperature is 10 DEG C, until not having
Until having anthocyanidin outflow；If still there is anthocyanidin outflow, adds deionized water and continue ultrafiltration.
（8）By step（7）Efflux be added in styrene type low pole macroreticular resin chromatographic column, first with pure water or
Deionized water elutes, and removes not by the impurity of macroporous resin adsorption；Again with 60% ethanol elution so that anthocyanidin is eluted down
Come, eluent is anthocyanidin ethanol solution.
（9）The NF membrane that molecular cut off is 200Da is added in anthocyanidin ethanol solution, nanofiltration is carried out, removes solvent second
Alcohol, operating pressure 0.1MPa, temperature are 10 DEG C.When anthocyanidin volumes of aqueous ethanol is reduced to 1/10th of original volume,
Stop nanofiltration operation, respectively obtains trapped fluid（Anthocyanidin solution）And ethyl alcohol, ethyl alcohol can be recycled.
（10）Trapped fluid is dried using lyophilization or spray drying process.First, by solution precooling to 0 DEG C with
Under.Then, it is less than or equal to 50kPa in vacuum degree, under the conditions of -20 DEG C, freeze-drying to moisture content of finished products content≤5%.Or
110 DEG C of conditions, solution is spray-dried, until moisture content of finished products content≤5%.Obtain the Black Box Tracing anthocyanidin powder of aubergine
End, the wherein content of anthocyanidin are 60%.
Not only low energy consumption but also has good environment friendly for the separation method of the present invention.The present invention mainly uses water to make
For extractant, biological enzymolysis centrifuges large granular impurity in removal solution, ultrafiltration impurity elimination, macroporous resin purification, ethyl alcohol conduct
Eluant, eluent, nanofiltration retention technology isolate ethyl alcohol.This method can be effectively saved energy consumption, and do not generate and any toxic have
Evil substance, the eluant, eluent of use --- ethyl alcohol can be recycled.
The extraction and purification process of the present invention preferably protects anthocyanidin not to be destroyed.In whole process of the present invention, use
Ultramicro grinding；Medium temperature（20~50 DEG C）Enzymolysis and extraction；High speed centrifugation impurity elimination；Ultra-filtration and separation purifies；Macroporous resin purification；Nanofiltration
Concentration and recycling ethyl alcohol；Freeze-drying is dried, and is farthest retained anthocyanidin and is not destroyed.
Embodiment two：The method for extraction and purification of anthocyanidin in a kind of Black Box Tracing pomace
（1）Fresh Black Box Tracing is beaten, pulp is obtained.
（2）Pulp is detached fruit juice with black slag using slurry slag separator or centrifuge.
（3）Pomace is added in vibration type ultramicro disintegrator, after the deionized water for adding 10 times of volumes, ultramicro grinding 40 is divided
Clock, until the average particle size of material is more than 300 mesh.
（4）Pomace after ultramicro grinding is added in enzymatic vessel, with the pH to 4.5 of citric acid regulating solution；Temperature 25~
Under the conditions of 55 DEG C, the pectase and 0.2% cellulase of solution quality 0.3% is added, is uniformly mixed, digests 6 hours, until in solution
Until there is no pectin to be detected, enzymolysis solution is obtained.
（5）Enzymolysis solution is used into separation decanter centrifuge, is centrifuged under 5000r/min rotating speeds, removal is most of
Residue obtains thick clear solution.
（6）Clear solution is used into self-cleaning type disk centrifuge, is centrifuged under 13000r/min rotating speeds, removal is remaining
A small amount of residue, obtain clear solution.
（7）Clear solution is added in the ultrafiltration apparatus that molecular cut off is 10kDa, carries out ultrafiltration, further remove residual
Deposit the big molecular impurities such as a small amount of polysaccharide, protein, cellulose, pectin.Control pressure is 0.7MPa, and temperature is 50 DEG C, until
Until there is no anthocyanidin outflow；If still there is anthocyanidin outflow, adds deionized water and continue ultrafiltration.
（8）By step（7）Efflux be added in styrene type non-polar macroporous resin chromatographic column, first with pure water or
Deionized water elutes, and removes not by the impurity of macroporous resin adsorption；Again with 90% ethanol elution so that anthocyanidin is eluted down
Come, eluent is anthocyanidin ethanol solution.
（9）The NF membrane that molecular cut off is 300Da is added in anthocyanidin ethanol solution, nanofiltration is carried out, removes solvent second
Alcohol, operating pressure 0.7MPa, temperature are 50 DEG C.When anthocyanidin volumes of aqueous ethanol is reduced to 1/10th of original volume,
Stop nanofiltration operation, respectively obtains trapped fluid（Anthocyanidin solution）And ethyl alcohol, ethyl alcohol can be recycled.
（10）Trapped fluid is dried using lyophilization or spray drying process.First, by solution precooling to 0 DEG C with
Under.Then, it is less than or equal to 50kPa in vacuum degree, under the conditions of -40 DEG C, freeze-drying to moisture content of finished products content≤5%.Or
120 DEG C of conditions, solution is spray-dried, until moisture content of finished products content≤5%.Obtain the Black Box Tracing anthocyanidin powder of aubergine
End, the wherein content of anthocyanidin are 90%.
The above embodiments merely illustrate the technical concept and features of the present invention, and its object is to allow person skilled in the art
Scholar cans understand the content of the present invention and implement it accordingly, and it is not intended to limit the scope of the present invention.It is all according to the present invention
Equivalent change or modification made by Spirit Essence, should be covered by the protection scope of the present invention.
Claims (6)
1. the method for extraction and purification of anthocyanidin in a kind of Black Box Tracing pomace, it is characterised in that：Include the following steps：
（1）Black Box Tracing is beaten to obtain pulp；
（2）Fruit juice in the pulp is detached with pomace；
（3）The pomace is added in vibration type ultramicro disintegrator, the deionization that volume is 2~10 times of the pomace is then added
After water, the average particle size of ultramicro grinding 30-40 minutes to material is greater than or equal to 300 mesh；
（4）Pomace after ultramicro grinding is added in enzymatic vessel, with the pH value of citric acid regulating solution to 2.5~4.5；Next
Under the conditions of temperature is 25~55 DEG C, pectase and concentration of polymer solution that addition concentration of polymer solution is 0.01~0.3% are
0.01~0.2% cellulase is uniformly mixed, and until not having pectin to be detected in solution, obtains enzymolysis solution；
（5）The enzymolysis solution is centrifuged under 3000-5000r/min rotating speeds, obtains thick clear solution；
（6）The thick clear solution is centrifuged under 5000-13000r/min rotating speeds, obtains clear solution；
（7）The clear solution is added in the ultrafiltration apparatus that molecular cut off is 3-10kDa and carries out ultrafiltration, removes the miscellaneous of remaining
Matter；Control pressure is 0.1~0.7MPa, and temperature is 10~50 DEG C, until without anthocyanidin outflow；
（8）By step（7）Efflux styrene type non-polar macroporous resin or styrene type low pole macroreticular resin is added
It in chromatographic column, is first eluted, is removed not by the impurity of macroporous resin adsorption with pure water or deionized water；It is with volumetric concentration again
60%~90% ethanol elution so that anthocyanidin is eluted, and eluent is anthocyanidin ethanol solution；
（9）The NF membrane that molecular cut off is 200-300Da is added in anthocyanidin ethanol solution, nanofiltration is carried out, removes solvent second
Alcohol, operating pressure is 0.1~0.7MPa, temperature is 10~50 DEG C；When anthocyanidin volumes of aqueous ethanol is reduced to original volume
1/10th, stop nanofiltration and operates to obtain anthocyanidin solution；
（10）Anthocyanidin solution is dried up to product using vacuum freeze-drying method or spray drying process.
2. the method for extraction and purification of anthocyanidin in Black Box Tracing pomace according to claim 1, it is characterised in that：Institute
It is the Black Box Tracing that thaws after fresh Black Box Tracing or freezing to state Black Box Tracing.
3. the method for extraction and purification of anthocyanidin in Black Box Tracing pomace according to claim 1, it is characterised in that：It adopts
The fruit juice in the pulp is detached with pomace with slurry slag separator or centrifuge.
4. the method for extraction and purification of anthocyanidin in Black Box Tracing pomace according to claim 1, it is characterised in that：
Step（7）In, when still there is anthocyanidin outflow, adds deionized water and continue ultrafiltration.
5. the method for extraction and purification of anthocyanidin in Black Box Tracing pomace according to claim 1, it is characterised in that：Institute
State vacuum freeze-drying method refer to by anthocyanidin solution precooling to 0 DEG C hereinafter, then vacuum degree be less than or equal to 50kPa ,-
Under the conditions of 20~-40 DEG C, freeze-drying to moisture content of finished products content≤5%.
6. the method for extraction and purification of anthocyanidin in Black Box Tracing pomace according to claim 1, it is characterised in that：Institute
It refers to that anthocyanidin solution is spray-dried to moisture content of finished products content≤5% in 110 ~ 120 DEG C of conditions to state spray drying process；It obtains purple
Red Black Box Tracing anthocyanidin powder, the wherein content of anthocyanidin are 60-90%.
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| CN113336734B (en) * | 2021-06-21 | 2022-06-03 | 秦皇岛惠恩生物技术有限公司 | Method for extracting procyanidine from aronia melanocarpa fruits |
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| CN105949163A (en) | 2016-09-21 |
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Address after: 214536, No. 1, Yao Dong Road, Xinqiao Town, Taizhou City, Jiangsu, Jingjiang Patentee after: Jiangsu Wisteria Ecological Technology Co., Ltd Address before: 214537, No. 1, Yao Dong Road, Xinqiao Town, Jiangsu, Taizhou Patentee before: Jiangsu Wisteria Garden Green Engineering Co., Ltd. |
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