CN107496465A - Compound based on ball algae extract and preparation method thereof - Google Patents
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- A61K36/05—Chlorophycota or chlorophyta (green algae), e.g. Chlorella
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- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
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- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
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Abstract
The invention discloses a kind of compound based on ball algae extract, composition and its parts by weight are 3 ~ 6 parts of 20 ~ 40 parts of ball algae extract, 50 ~ 70 parts of cornstarch, 5 ~ 10 parts of Arabic gum, 20 ~ 30 parts of CMS, 15 ~ 30 parts of vitamin C and folic acid.Its preparation method includes freeze thawing supersound process, deproteinized, dialysis and alcohol precipitation, isolates and purifies preparation with compound.Have the beneficial effect that：The property of ball algae extract is stable in the compound being prepared, and each composition mutually acts synergistically in compound, can improve the amoeboid movement speed of leucocyte, shortens the time that leucocyte reaches tissue where pathogen from blood；The ability of the cytokine profiles such as phagocytic activity and secreting leukocytes mesonium, interferon, the TNF of leucocyte can be improved simultaneously.After long-term use of above-mentioned polysaccharide, flu frequency declines, and body immunity is significantly improved.
Description
Technical field
The present invention relates to compound preparing technical field, specifically a kind of compound and its preparation based on ball algae extract
Method.
Background technology
Polysaccharide is a kind of large biological molecule being widely present in animal cell membrane, plant and microorganism wall, is structure
One of four big base substances into life.Research in recent decades finds that polysaccharide has complicated biology and physiologically active, especially
It is in recent ten years with sides such as countries in the world scientist isolates and purifies to plant polyose, structural analysis, immunology and pharmacology
Face research deepens continuously, and find the cheap plant polyose source of a kind of resource relative abundance, cost turns into food circle, medical field
The focus of concern.Ocean area is vast, seaweed plant abundance, wherein except rich in good protein, amino acid, cellulose
And outside mineral element, also containing multiple biological activities polysaccharide, and marine alga living environment is special, algal polysaccharides have unique life
Reason activity.The attention of countries in the world is increasingly caused as medical treatment, health treatment, algal polysaccharides just progressively turn into biological polyoses
One of main source.
Chlorella (ChlorellapyrenoidosaFACHB5) is a kind of general natural disposition unicellular alga.Chlorella has life
State distribution is wide, can fast-growth and breeding, be easy to the advantages that culture, be that uniquely can increase by 4 times in 20h in animals and plants on the earth
Biology, there is very high application value.Chlorella has been widely used in animal feed, food additives, medicine system at present
Agent, cosmetics etc..
Prior art such as Authorization Notice No. is CN104311685B Chinese invention patent, discloses a kind of spirulina polysaccharide
And its extracting method, comprise the following steps：After spirulina powder suspension multigelation broken wall, centrifugation obtains clasmatosis liquid 1
And cell fragment；It is 50% that ammonium sulfate to its saturation degree is added in clasmatosis liquid 1, and after salt precipitation, centrifugation removes algae courage
Albumen, supernatant is obtained, supernatant and cell fragment are mixed, obtain clasmatosis liquid 2；Spiral is obtained using Hot water extraction
Polysaccharides crude extract；Crude extract is added in ethanol/ammonium sulfate double-aqueous phase system and extracted, obtains the spiral with higher degree
The lower phase solution of polysaccharides；Lower phase solution is after desalination of dialysing, with SephadexG-150 chromatographic columns and DEAESephadexA-50
Anion-exchange column elutes, and obtains pure spirulina polysaccharide solution；After freeze-drying, pure spirulina polysaccharide finished product is prepared into.It is above-mentioned
Method preparation process is cumbersome, and active polysaccharide is more easily damaged in extraction process, and obtained product quality is not high.
The content of the invention
It is an object of the invention to provide a kind of preparation process is simple, raw material sources are wide, cheap, the high base of recovery rate
Ball polysaccharides property is stable in compound of ball algae extract and preparation method thereof, obtained compound, long-term use of above-mentioned
Compound can significantly improve immunity.
The present invention is directed to the problem of being mentioned in background technology, and the technical scheme taken is：Compounding based on ball algae extract
Thing, composition and its parts by weight are 20 ~ 40 parts of ball algae extract, 50 ~ 70 parts of cornstarch, 5 ~ 10 parts of Arabic gum, carboxymethyl shallow lake
3 ~ 6 parts of 20 ~ 30 parts of powder, 15 ~ 30 parts of vitamin C and folic acid.
Based on the preparation method of ball algae extract, including freeze thawing-supersound process, deproteinized, dialysis and alcohol precipitation, isolate and purify
With the preparation of compound.The property of ball algae extract is stable in compound made from the above method, and respectively into split-phase in compound
Mutually synergy, the amoeboid movement speed of leucocyte can be improved, shorten tissue where leucocyte reaches pathogen from blood
Time；The various kinds of cell such as phagocytic activity and secreting leukocytes mesonium, interferon, the TNF of leucocyte can be improved simultaneously
The ability of the factor.After long-term use of above-mentioned polysaccharide, flu frequency declines, and body immunity is significantly improved.Compound carries
The effect of high immunity is superior to any single composition in formula.Purification procedures are the method using gel chromatography to thick more
Sugar is isolated and purified, and eluent is the aqueous solution dissolved with active peptides.Above-mentioned gel chromatography elution elution liquid measure used is few,
It is few to be improved the function polysaccharide loss amount of immunity, the good separating effect of elution.
The content of active peptides is 1 ~ 3g/L, and the amino acid sequence of active peptides is
SCASRCKSRCRACRRCKGYAAVSFGRCRCICKCFRC.Above-mentioned active peptides can significantly improve the K values of function polysaccharide, to it
The K values of his polysaccharide influence less, so as to effectively isolate and purify function polysaccharide.Function polysaccharide property is unstable, easily by outer
Boundary factor influences and inactivated, above-mentioned active peptides can preferable defencive function polysaccharide activity.
Gel chromatography elution flow rate is 0.2 ~ 0.7mL/min, collects the eluent for having absorption at 620nm, is concentrated and cold
Jelly is dried to obtain ball algae extract.Function polysaccharide has a stronger absorption at 620nm, and other polysaccharide are here without absorption, according to
This characteristic is enriched with to function polysaccharide.
Freeze thawing-ultrasound treatment step adds phosphate buffer to take chlorella algal gel, repeatedly ultrasonic freeze thawing 1 ~ 4 time, centrifugation
Supernatant is taken, ultrasonic power is 400 ~ 700W, and each ultrasonication time is 4 ~ 7min.Made using cavitation caused by ultrasonic wave
Accelerate the broken of chlorella cells wall with, mechanism, promote the dissolution of intracellular organic matter, improve the recovery rate of polysaccharide.
Phosphate buffer is 12 ~ 18 with algal gel mass ratio, and buffer concentration is 0.04 ~ 0.08mol/L, and pH is 7.0 ~ 8.5.
Solubility of the function polysaccharide in above-mentioned phosphate buffer is high, and can preferably keep the activity of function polysaccharide.
Deproteinized step is adds 2 ~ 5% solution of trichloroacetic acid in supernatant, until no longer there is white precipitate, from
The heart takes supernatant.Trichloroacetic acid is a kind of organic acid, can make the protein denaturation precipitation in solution.Above method operation letter
Single, result stabilization, reaction are gentle, can ensure the activity of polysaccharide well in removing protein.
Dialyse and alcohol precipitation step is that supernatant is placed in into 40 ~ 60h of dialysis in the bag filter that specification is 200 ~ 400Da, add 2
The ethanol of ~ 5 times of volumes, stands 2 ~ 4h, centrifuging and taking precipitation, and low temperature drying obtains Thick many candies.Ethanol, ethanol are added in polysaccharide solution
The hydrated sheath of polysaccharide surface is destroyed, solubility of the polysaccharide in water is substantially reduced, plays the effect of separating-purifying, meanwhile, polysaccharide
Activity be not destroyed.
The preparation process of compound is to be formed sediment by formula ratio mixing ball algae extract, cornstarch, Arabic gum, carboxymethyl
Powder, vitamin C and folic acid, are made into tablet, obtain the compound based on ball algae extract.
Compared with prior art, the advantage of the invention is that：1）The inventive method is simple to operate, with short production cycle, required
Equipment it is few, raw material sources are wide, and production cost is low；2）The property of ball algae extract is stable in the compound being prepared, and multiple
Mutually acted synergistically with each composition in thing, the amoeboid movement speed of leucocyte can be improved, shortened leucocyte and disease is reached from blood
The time of tissue where substance；The phagocytic activity and secreting leukocytes mesonium, interferon, neoplasm necrosis of leucocyte can be improved simultaneously
The ability of the cytokine profiles such as the factor.After long-term use of above-mentioned polysaccharide, flu frequency declines, and body immunity obtains significantly
Improve；3）It is more that the activity that amino acid sequence is SCASRCKSRCRACRRCKGYAAVSFGRCRCICKCFRC is added in eluent
Peptide.Above-mentioned active peptides can significantly improve the K values of function polysaccharide, the K values of other polysaccharide be influenceed less, so as to effectively by work(
Can separation of polysaccharides purifying.Function polysaccharide property is unstable, is easily influenceed and inactivated by extraneous factor, and above-mentioned active peptides can be preferable
The activity of defencive function polysaccharide.
Specific embodiment
The present invention program is described further below by embodiment：
Embodiment 1：
Based on the compound of ball algae extract, composition and its parts by weight are 30 parts of ball algae extract, 60 parts of cornstarch, Arab
4 parts of 8 parts of glue, 24 parts of CMS, 18 parts of vitamin C and folic acid.
Based on the preparation method of ball algae extract, including freeze thawing-supersound process, deproteinized, dialysis and alcohol precipitation, isolate and purify
With the preparation of compound.The property of ball algae extract is stable in compound made from the above method, and respectively into split-phase in compound
Mutually synergy, the amoeboid movement speed of leucocyte can be improved, shorten tissue where leucocyte reaches pathogen from blood
Time；The various kinds of cell such as phagocytic activity and secreting leukocytes mesonium, interferon, the TNF of leucocyte can be improved simultaneously
The ability of the factor.After long-term use of above-mentioned polysaccharide, flu frequency declines, and body immunity is significantly improved.Compound carries
The effect of high immunity is superior to any single composition in formula.Purification procedures are the method using gel chromatography to thick more
Sugar is isolated and purified, and eluent is the aqueous solution dissolved with active peptides.Above-mentioned gel chromatography elution elution liquid measure used is few,
It is few to be improved the function polysaccharide loss amount of immunity, the good separating effect of elution.
The content of active peptides is 3g/L, and the amino acid sequence of active peptides is
SCASRCKSRCRACRRCKGYAAVSFGRCRCICKCFRC.Above-mentioned active peptides can significantly improve the K values of function polysaccharide, to it
The K values of his polysaccharide influence less, so as to effectively isolate and purify function polysaccharide.Function polysaccharide property is unstable, easily by outer
Boundary factor influences and inactivated, above-mentioned active peptides can preferable defencive function polysaccharide activity.
Gel chromatography elution flow rate is 0.6mL/min, collects the eluent for having absorption at 620nm, concentrates and freezes dry
It is dry to obtain ball algae extract.Function polysaccharide has a stronger absorption at 620nm, and other polysaccharide are here without absorption, according to this
Characteristic is enriched with to function polysaccharide.
Freeze thawing-ultrasound treatment step adds phosphate buffer to take chlorella algal gel, repeatedly ultrasonic freeze thawing 1 ~ 4 time, centrifugation
Supernatant, ultrasonic power 500W are taken, each ultrasonication time is 7min.Utilize cavitation, machinery caused by ultrasonic wave
Effect accelerates the broken of chlorella cells wall, promotes the dissolution of intracellular organic matter, improves the recovery rate of polysaccharide.
Phosphate buffer is 16 with algal gel mass ratio, buffer concentration 0.06mol/L, pH 7.5.Function polysaccharide is upper
The solubility stated in phosphate buffer is high, and can preferably keep the activity of function polysaccharide.
Deproteinized step is adds 3% solution of trichloroacetic acid in supernatant, until no longer there is white precipitate, centrifuges
Take supernatant.Trichloroacetic acid is a kind of organic acid, can make the protein denaturation precipitation in solution.The above method is simple to operate,
As a result stablize, react gentle, the activity of polysaccharide can be ensured well in removing protein.
Dialyse and alcohol precipitation step is that supernatant is placed in the bag filter that specification is 400Da into the 50h that dialyses, add 4 times of volumes
Ethanol, stand 2h, centrifuging and taking precipitation, low temperature drying obtains Thick many candies.Ethanol is added in polysaccharide solution, ethanol destroys polysaccharide table
The hydrated sheath in face, substantially reduce solubility of the polysaccharide in water, play the effect of separating-purifying, meanwhile, polysaccharide activity not by
Destroy.
The preparation process of compound is to be formed sediment by formula ratio mixing ball algae extract, cornstarch, Arabic gum, carboxymethyl
Powder, vitamin C and folic acid, are made into tablet, obtain the compound based on ball algae extract.
Embodiment 2：
Based on the compound of ball algae extract, composition and its parts by weight are 36 parts of ball algae extract, 60 parts of cornstarch, Arab
5 parts of 7 parts of glue, 30 parts of CMS, 7 parts of Catergen and folic acid.
The preparation method of compound based on ball algae extract, comprises the following steps：
1）Freeze thawing-supersound process：Take 4g chlorella algal gels, add 60g phosphate buffers, repeatedly ultrasonic freeze thawing 2 times, in rotating speed
30min is centrifuged under the conditions of 6000r/min and takes supernatant.Ultrasonic power is 600W, and each ultrasonication time is 6min.Phosphoric acid delays
Fliud flushing concentration is 0.06mol/L, pH 7.5；
2）Deproteinized：3% solution of trichloroacetic acid is added in supernatant, until no longer there is white precipitate, centrifuging and taking supernatant
Liquid；
3）Dialysis and alcohol precipitation：Supernatant is placed in the bag filter that specification is 300Da the 48h that dialyses, adds the ethanol of 3 times of volumes,
3h is stood, centrifuging and taking precipitation, 1h is dried at 25 DEG C, under the conditions of 100KPa and obtains Thick many candies powder；
4）Isolate and purify：Thick many candies are isolated and purified using the method for gel chromatography, eluent is dissolved with active peptides
The aqueous solution.The content of active peptides is 2g/L, and the amino acid sequence of active peptides is
SCASRCKSRCRACRRCKGYAAVSFGRCRCICKCFRC.Gel chromatography elution flow rate is 0.5mL/min, is collected in 620nm
There is the eluent of absorption at place, concentrates and is freeze-dried to obtain ball algae extract, and chlorella extract yield is 6.718%；
5）The preparation of compound：By formula ratio mixing ball algae extract, cornstarch, Arabic gum, CMS, vitamin
C and folic acid, are made into tablet, obtain the compound based on ball algae extract.Ball algae carries in compound made from the above method
Take the property of thing stable, and each composition mutually acts synergistically in compound, can improve the amoeboid movement speed of leucocyte, shortens white
Cell reaches the time of tissue where pathogen from blood；The phagocytic activity and secretion leucocyte that leucocyte can be improved simultaneously are situated between
The ability of the cytokine profiles such as element, interferon, TNF.After long-term use of above-mentioned polysaccharide, flu frequency declines, people
Body immunity is significantly improved.The effect that compound improves immunity is superior to any single composition in formula.
Embodiment 3：
Based on the compound of ball algae extract, composition and its parts by weight are 30 parts of ball algae extract, 52 parts of cornstarch, Arab
4 parts of 6 parts of glue, 22 parts of CMS, 3 parts of Catergen and folic acid.
The preparation method of compound based on ball algae extract, comprises the following steps：
1）Freeze thawing-supersound process：Take 6g chlorella algal gels, add 35g phosphate buffers, repeatedly ultrasonic freeze thawing 2 times, in rotating speed
30min is centrifuged under the conditions of 6000r/min and takes supernatant.Ultrasonic power is 800W, and each ultrasonication time is 4min.Phosphoric acid delays
Fliud flushing concentration is 0.06mol/L, pH 7.5；
2）Deproteinized：4% solution of trichloroacetic acid is added in supernatant, until no longer there is white precipitate, centrifuging and taking supernatant
Liquid；
3）Dialysis and alcohol precipitation：Supernatant is placed in the bag filter that specification is 300Da the 48h that dialyses, adds the ethanol of 3 times of volumes,
3h is stood, centrifuging and taking precipitation, 1.5h is dried at 30 DEG C, under the conditions of 200KPa and obtains Thick many candies powder；
4）Isolate and purify：Thick many candies are isolated and purified using the method for gel chromatography, eluent is dissolved with active peptides
The aqueous solution.The content of active peptides is 2g/L, and the amino acid sequence of active peptides is
SCASRCKSRCRACRRCKGYAAVSFGRCRCICKCFRC.Gel chromatography elution flow rate is 0.5mL/min, is collected in 620nm
There is the eluent of absorption at place, concentrates and is freeze-dried to obtain ball algae extract, and chlorella extract yield is 6.314%；
5）The preparation of compound：By formula ratio mixing ball algae extract, cornstarch, Arabic gum, CMS, vitamin
C and folic acid, are made into tablet, obtain the compound based on ball algae extract.Ball algae carries in compound made from the above method
Take the property of thing stable, and each composition mutually acts synergistically in compound, can improve the amoeboid movement speed of leucocyte, shortens white
Cell reaches the time of tissue where pathogen from blood；The phagocytic activity and secretion leucocyte that leucocyte can be improved simultaneously are situated between
The ability of the cytokine profiles such as element, interferon, TNF.After long-term use of above-mentioned polysaccharide, flu frequency declines, people
Body immunity is significantly improved.The effect that compound improves immunity is superior to any single composition in formula.
Routine operation in the operating procedure of the present invention is well known to those skilled in the art, herein without repeating.
Technical scheme is described in detail embodiment described above, it should be understood that it is described above only
For the specific embodiment of the present invention, it is not intended to limit the invention, all any modifications made in the spirit of the present invention,
Supplement or similar fashion replacement etc., should be included in the scope of the protection.
SEQUENCE LISTING
<110>Lanxi City silence bio tech ltd
<120>Compound based on ball algae extract and preparation method thereof
<130> 1
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 36
<212> PRT
<213>It is artificial synthesized
<400> 1
Ser Cys Ala Ser Arg Cys Lys Ser Arg Cys Arg Ala Cys Arg Arg Cys
1 5 10 15
Lys Gly Tyr Ala Ala Val Ser Phe Gly Arg Cys Arg Cys Ile Cys Lys
20 25 30
Cys Phe Arg Cys
35
Claims (8)
1. the compound based on ball algae extract, it is characterised in that composition and its parts by weight are 20 ~ 40 parts of ball algae extract, corn
3 ~ 6 parts of 50 ~ 70 parts of starch, 5 ~ 10 parts of Arabic gum, 20 ~ 30 parts of CMS, 15 ~ 30 parts of vitamin C and folic acid.
2. based on the preparation method of ball algae extract, including freeze thawing-supersound process, deproteinized, dialysis and alcohol precipitation, isolate and purify and
The preparation of compound, it is characterised in that：Described purification procedures are that Thick many candies are divided using the method for gel chromatography
From purifying, eluent is the aqueous solution dissolved with active peptides.
3. the preparation method of the compound according to claim 2 based on ball algae extract, it is characterised in that：Described work
The content of property polypeptide is 1 ~ 3g/L, and the amino acid sequence of active peptides is
SCASRCKSRCRACRRCKGYAAVSFGRCRCICKCFRC。
4. according to the preparation method of the compound based on ball algae extract described in claim 2, it is characterised in that：Described gel
Chromatographic elution flow velocity is 0.2 ~ 0.7mL/min, collects the eluent for having absorption at 620nm, concentrates and is freeze-dried to obtain ball
Algae extract.
5. the preparation method of the compound according to claim 2 based on ball algae extract, it is characterised in that：Described jelly
Melt-ultrasound treatment step to take chlorella algal gel, add phosphate buffer, repeatedly ultrasonic freeze thawing 1 ~ 4 time, centrifuging and taking supernatant,
Ultrasonic power is 400 ~ 700W, and each ultrasonication time is 4 ~ 7min.
6. the preparation method according to claim 5 based on ball algae extract, it is characterised in that：Described phosphate buffer
It is 12 ~ 18 with algal gel mass ratio, buffer concentration is 0.04 ~ 0.08mol/L, and pH is 7.0 ~ 8.5.
7. according to the preparation method of the compound based on ball algae extract described in claim 2, it is characterised in that：Described removes egg
White step is adds 2 ~ 5% solution of trichloroacetic acid in supernatant, until no longer there is white precipitate, centrifuging and taking supernatant.
8. according to the preparation method of the compound based on ball algae extract described in claim 2, it is characterised in that：Described dialysis
And alcohol precipitation step is that supernatant is placed in into 40 ~ 60h of dialysis in the bag filter that specification is 200 ~ 400Da, adds the second of 2 ~ 5 times of volumes
Alcohol, stands 2 ~ 4h, centrifuging and taking precipitation, and low temperature drying obtains Thick many candies.
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| Application Number | Priority Date | Filing Date | Title |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107353357A (en) * | 2017-07-01 | 2017-11-17 | 浦江县泰如食品科技有限公司 | For improving the ball algae extract of immunity |
| CN107875179A (en) * | 2017-10-26 | 2018-04-06 | 海南三元星生物科技股份有限公司 | One kind mixing algae extract and its preparation method and application |
| CN108753623A (en) * | 2018-06-12 | 2018-11-06 | 兰溪市沉默生物科技有限公司 | A kind of chlorella store method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105085697A (en) * | 2015-08-05 | 2015-11-25 | 都宝君 | Chlorella polysaccharide extraction method |
| CN106389367A (en) * | 2016-11-16 | 2017-02-15 | 杭州朱养心药业有限公司 | Perampanel coated tablet pharmaceutical composition |
-
2017
- 2017-06-21 CN CN201710475601.1A patent/CN107496465A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105085697A (en) * | 2015-08-05 | 2015-11-25 | 都宝君 | Chlorella polysaccharide extraction method |
| CN106389367A (en) * | 2016-11-16 | 2017-02-15 | 杭州朱养心药业有限公司 | Perampanel coated tablet pharmaceutical composition |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107353357A (en) * | 2017-07-01 | 2017-11-17 | 浦江县泰如食品科技有限公司 | For improving the ball algae extract of immunity |
| CN107875179A (en) * | 2017-10-26 | 2018-04-06 | 海南三元星生物科技股份有限公司 | One kind mixing algae extract and its preparation method and application |
| CN107875179B (en) * | 2017-10-26 | 2020-10-30 | 海南三元星生物科技股份有限公司 | Mixed algae extract and preparation method and application thereof |
| CN108753623A (en) * | 2018-06-12 | 2018-11-06 | 兰溪市沉默生物科技有限公司 | A kind of chlorella store method |
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