CN106974284A - A kind of chlamydomonas ethanol extract and preparation method and application - Google Patents
A kind of chlamydomonas ethanol extract and preparation method and application Download PDFInfo
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- 241000195585 Chlamydomonas Species 0.000 title claims abstract description 67
- 239000000469 ethanolic extract Substances 0.000 title claims abstract description 46
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 32
- 241000195493 Cryptophyta Species 0.000 claims abstract description 21
- 239000006228 supernatant Substances 0.000 claims abstract description 19
- 238000001556 precipitation Methods 0.000 claims abstract description 18
- 239000003963 antioxidant agent Substances 0.000 claims abstract description 10
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 10
- 235000006708 antioxidants Nutrition 0.000 claims abstract description 10
- 238000002137 ultrasound extraction Methods 0.000 claims abstract description 10
- 239000003814 drug Substances 0.000 claims abstract description 8
- 238000005119 centrifugation Methods 0.000 claims abstract 2
- 239000007788 liquid Substances 0.000 claims description 9
- 239000000047 product Substances 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 7
- 239000000155 melt Substances 0.000 claims description 5
- 230000036541 health Effects 0.000 claims description 3
- 238000001291 vacuum drying Methods 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims 1
- 241000244203 Caenorhabditis elegans Species 0.000 abstract description 24
- 230000036542 oxidative stress Effects 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 9
- 238000001727 in vivo Methods 0.000 abstract description 9
- 229910052760 oxygen Inorganic materials 0.000 abstract description 9
- 239000001301 oxygen Substances 0.000 abstract description 9
- 230000003712 anti-aging effect Effects 0.000 abstract description 8
- 235000013402 health food Nutrition 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 239000003960 organic solvent Substances 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 10
- 238000012360 testing method Methods 0.000 description 7
- 230000004083 survival effect Effects 0.000 description 6
- 230000012010 growth Effects 0.000 description 5
- 230000032683 aging Effects 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- FIKAKWIAUPDISJ-UHFFFAOYSA-L paraquat dichloride Chemical compound [Cl-].[Cl-].C1=C[N+](C)=CC=C1C1=CC=[N+](C)C=C1 FIKAKWIAUPDISJ-UHFFFAOYSA-L 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 239000002244 precipitate Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000010257 thawing Methods 0.000 description 3
- 238000002604 ultrasonography Methods 0.000 description 3
- 239000002028 Biomass Substances 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000003064 anti-oxidating effect Effects 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 230000000243 photosynthetic effect Effects 0.000 description 2
- 239000006187 pill Substances 0.000 description 2
- 230000009758 senescence Effects 0.000 description 2
- 240000002900 Arthrospira platensis Species 0.000 description 1
- 235000016425 Arthrospira platensis Nutrition 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 1
- 241000195633 Dunaliella salina Species 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000003225 biodiesel Substances 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000003495 flagella Anatomy 0.000 description 1
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- 239000013505 freshwater Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 239000004530 micro-emulsion Substances 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 239000003182 parenteral nutrition solution Substances 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 239000001508 potassium citrate Substances 0.000 description 1
- 229960002635 potassium citrate Drugs 0.000 description 1
- QEEAPRPFLLJWCF-UHFFFAOYSA-K potassium citrate (anhydrous) Chemical compound [K+].[K+].[K+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O QEEAPRPFLLJWCF-UHFFFAOYSA-K 0.000 description 1
- 235000011082 potassium citrates Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 229940082787 spirulina Drugs 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
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- 239000003826 tablet Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
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- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
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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
- A61K36/02—Algae
- A61K36/05—Chlorophycota or chlorophyta (green algae), e.g. Chlorella
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- 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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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Chemical & Material Sciences (AREA)
- Alternative & Traditional Medicine (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention belongs to field of marine biotechnology, there is provided a kind of chlamydomonas ethanol extract and preparation method and application.The chlamydomonas ethanol extract that the present invention is provided is by by the fresh wet algae multigelation of chlamydomonas, centrifugation goes after supernatant to take precipitation plus ethanol progress ultrasonic extraction to be made, the time-to-live of Caenorhabditis elegans and the average life span of Caenorhabditis elegans under extension natural conditions under oxidative stress conditions can significantly be extended, simultaneously, it can promote activity in vivo oxygen radical (ROS) removing, significantly reduce activity in vivo oxygen radical (ROS) level, with significant anti-oxidant and anti-aging effects, it can be applied to prepare and there is anti-oxidant and/or anti-senescence function health food and medicine.The preparation method that the present invention is provided is simple and easy to apply, and organic solvent consumption is few, is adapted to industrialized production.
Description
Technical field
The invention belongs to field of marine biotechnology, more particularly to a kind of chlamydomonas ethanol extract and preparation method thereof with should
With.
Background technology
Microalgae be individually present in the water body environments such as a class Yu Haiyang, lake, river or into chain, bulk exist it is slender
Born of the same parents' microalgae.Microalgae has great bio-diversity, is widely distributed, and about has 200000~800000 on the earth
The microalgae of different classes is planted, wherein about 35000 kinds have been described, it is new more than what 15000 kinds produced from algal biomass
Compound has carried out chemical identification.
Microalgae have distribution wide and be not take up arable land, biomass is big, photosynthetic efficiency is high, strong environmental adaptability, growth week
The features such as phase is short, fat content is high and environment-friendly, therefore paid attention in the exploitation of biodiesel, current major part microalgae
Application and in terms of research is all concentrated mainly on efficient oil-producing.Further, since microalgae is nutritious, rich in micro- and all kinds of
Bioactive substance, therefore, it also has in terms of medicine, health products, cosmetics, aquaculture feed and feed addictive
Wide application prospect.But application study of the current microalgae in terms of medicine, health products is less, and the research in terms of these is current
The several frequently seen microalgaes such as spirulina, chlorella, Dunaliella salina and purple ball algae are concentrated mainly on, it is then relative for other microalgaes
Report is less.
Chlamydomonas Chlorophyta Chlorophyceae volvocales chlamydomonas section Chlamydomonas, frond is unicellular, and spherical or avette, front end has two
The isometric flagellum of bar, can move about.Chlamydomonas have the title of " green yeast ", has much in common, such as growth cycle letter with yeast
Single, growth is fast, and generation time is short, and monoclonal can be formed on flat board, Liquid Culture can be also carried out, with monoploid and diploid
Two kinds of form growths, can carry out tetrad analysis to the growth course of its gamete.Chlamydomonas be unique in that its in illumination and
It can be grown under dark condition, and carbon source and the photosynthetic carbonizable substance that the external world can be provided are separated, and are that only energy is special at present
The biology of conversion system is set up on ground to mitochondria, chloroplaset and core respectively.The research to chlamydomonas is concentrated mainly on bio at present
In terms of oil and biological hydrogen production, the bioactivity research of its anti-oxidant aspect is then rarely reported.Therefore active development has antioxygen
Change, the prevention and treatment to diseases associated with senescence are of great importance by the chlamydomonas extract of anti-aging effects.
The content of the invention
To solve problems of the prior art, the invention provides a kind of chlamydomonas ethanol extract.The present invention is provided
Chlamydomonas ethanol extract can significantly extend under oxidative stress conditions the time-to-live of Caenorhabditis elegans and extension nature bar
The average life span of Caenorhabditis elegans under part, meanwhile, it can promote activity in vivo oxygen radical (ROS) removing, significantly reduce
The level of activity in vivo oxygen radical (ROS), with significant anti-oxidant and anti-aging effects, can be applied to prepare has antioxygen
Change and/or the health food and medicine of anti-senescence function.
Technical scheme will be further described in detail below reflect and description.
A kind of chlamydomonas ethanol extract, its preparation method comprises the following steps：
S1, the wet algae of fresh chlamydomonas is taken, be to freeze 12 hours under conditions of -20 DEG C~-40 DEG C in temperature, room temperature melts 12
Hour, multigelation 3~5 times centrifuges 5~15min, by supernatant under conditions of rotating speed is 5000r/min~10000r/min
Liquid and precipitation are separated, and take precipitation standby；
S2, the EtOH Sonicate that addition volume fraction is 70%~99% into the precipitation described in step S1 are extracted 2~4 times,
5~15min is centrifuged under conditions of rotating speed is 5000r/min~10000r/min, supernatant and precipitation are separated, supernatant is taken
It is dried in vacuo, the one-level water of the product after vacuum drying is dissolved, chlamydomonas ethanol extract is produced.
Preferably, add ethanol by the solid-to-liquid ratio 1g/mL of wet algae and ethanol in step S2 and carry out ultrasonic extraction.
Preferably, the frequency of ultrasonic extraction is 35~40kHz in step S2, and temperature is 4 DEG C.
Preferably, the time of each ultrasonic extraction is 10~30min in step S2.
Chlamydomonas (Chlamydomonas sp.) of the present invention is provided by Inst. of Hydrobiology, Chinese Academy of Sciences, its
Numbering in Chinese Academy of Sciences's algae kind storehouse is FACHB-889.
The present inventor has carried out substantial amounts of experiment during studying microalgae, has been surprisingly found that to multigelation,
Remaining residue progress EtOH Sonicate, which extracts the chlamydomonas ethanol extract being made, after centrifuging and taking supernatant can significantly extend oxidation
The time-to-live of Caenorhabditis elegans and the average life span of Caenorhabditis elegans under extension natural conditions under stress conditions, meanwhile,
It can promote activity in vivo oxygen radical (ROS) removing, significantly reduce activity in vivo oxygen radical (ROS) level, have
Significant anti-oxidant and anti-aging effects, can be applied to prepare has anti-oxidant and/or anti-senescence function health food and medicine
Product.
Chlamydomonas ethanol extract of the present invention can be shared individually or with other active components, be used as solely or mainly active component
Applied to health food and medicine of the preparation with anti-oxidant and/or anti-senescence function function, the health food and medicine
Formulation is preferably tablet, capsule, parenteral solution, oral liquid, granule, pill, powder and pill etc..
Anti-oxidant, anti-aging effects of the present invention refer to anti-because occurring oxidative stress under pathology or physiological condition
Change that should be produced is the aging of principal character and the preventive and therapeutic effect of diseases associated with senescence.
Compared with prior art, the invention has the advantages that：
(1) the result of the test table tested in whole animal level with oxidative stress model and aging correlation model
It is bright, when the chlamydomonas ethanol extract that the present invention is provided can significantly extend the existence of Caenorhabditis elegans under oxidative stress conditions
Between, its survival rate is improved, while the average life span of Caenorhabditis elegans under natural conditions can also be extended, with significant anti-ageing
Old and antioxidation.
(2) the chlamydomonas ethanol extract that the present invention is provided can promote activity in vivo oxygen radical (ROS) removing, significantly drop
Low activity in vivo oxygen radical (ROS) level, with significant antioxidation.
(3) preparation method for the chlamydomonas ethanol extract that the present invention is provided has the cycle short, and organic solvent consumption is few, technique
Simply, the features such as safety non-toxic, stability and high efficiency, it is adapted to industrialized production.
Brief description of the drawings
Influence of Fig. 1 chlamydomonas ethanol extract to Caenorhabditis elegans survival rate under oxidative stress conditions.
Influence of Fig. 2 chlamydomonas ethanol extract to Caenorhabditis elegans average life span under natural conditions.
Influence of Fig. 3 chlamydomonas ethanol extract to ROS levels in Caenorhabditis elegans body.
Embodiment
Below by specific embodiment, the present invention is described in further detail.
Test method used in the embodiment of the present invention, is conventional method unless otherwise specified；Used material
Material, reagent, are the reagent and material commercially obtained unless otherwise specified.
In the embodiment of the present invention, chlamydomonas is provided by Inst. of Hydrobiology, Chinese Academy of Sciences, and it is in Chinese Academy of Sciences's fresh water
Numbering in algae kind storehouse is FACHB-889.
S Basal solution：Weigh 5.8gNaCl, 1.3g K2HPO4·3H2O and 6.0g KH2PO4It is placed in reagent bottle, plus
Enter 750mL deionized waters, be settled to 1L after being completely dissolved, sterilize 30min, be cooled to 40-50 DEG C it is standby.
S Medium solution：The S Basal solution after 1L sterilizings is taken, 1mL 5.0mg/mL cholesterol ethanol is added dropwise molten
Liquid, while add change to shake, makes it fully dissolve (solution switch to micro emulsion by water white transparency transparent in vain, no obvious sediment), then successively
Add 3mL 1M CaCl2Solution, 3mL 1M MgSO4, 10mL 1M potassium citrate solutions (pH=6.0) and 10mL trace elements are molten
Liquid, room temperature is standby.
The preparation of the chlamydomonas ethanol extract of embodiment 1
S1, the wet algae of fresh chlamydomonas is taken, be to freeze 12 hours under conditions of -40 DEG C in temperature, room temperature melts 12 hours, instead
Freeze thawing 4 times again, centrifuges 10min under conditions of rotating speed is 10000r/min, supernatant and precipitation is separated, take and precipitate standby；
S2, to add volume fraction by the solid-to-liquid ratio 1g/mL of wet algae and ethanol into the precipitation described in step S1 be 80%
EtOH Sonicate is extracted 3 times, and the frequency of ultrasonic extraction is 40kHz, and temperature is 4 DEG C, and the time of ultrasound is 25min every time, in rotating speed
To centrifuge 15min under conditions of 10000r/min, supernatant and precipitation are separated, take supernatant to be dried in vacuo, by vacuum
Dried product one-level water dissolves, and it is that (A represents that chlamydomonas is wet to 1gA/mL to adjust it relative to the concentration of chlamydomonas (wet algae) weight
Algae), produce chlamydomonas ethanol extract.
The preparation of the chlamydomonas ethanol extract of embodiment 2
S1, the wet algae of fresh chlamydomonas is taken, be to freeze 12 hours under conditions of -20 DEG C in temperature, room temperature melts 12 hours, instead
Freeze thawing 5 times again, centrifuges 15min under conditions of rotating speed is 5000r/min, supernatant and precipitation is separated, take and precipitate standby；
S2, to add volume fraction by the solid-to-liquid ratio 1g/mL of wet algae and ethanol into the precipitation described in step S1 be 70%
EtOH Sonicate is extracted 3 times, and the frequency of ultrasonic extraction is 35kHz, and temperature is 4 DEG C, and the time of ultrasound is 30min every time, in rotating speed
To centrifuge 15min under conditions of 8000r/min, supernatant and precipitation are separated, takes supernatant to be dried in vacuo, vacuum is done
Product after dry is dissolved with one-level water, and it is that (A represents that chlamydomonas is wet to 1gA/mL to adjust it relative to the concentration of chlamydomonas (wet algae) weight
Algae), produce chlamydomonas ethanol extract.
The preparation of the chlamydomonas ethanol extract of embodiment 3
S1, the wet algae of fresh chlamydomonas is taken, be to freeze 12 hours under conditions of -30 DEG C in temperature, room temperature melts 12 hours, instead
Freeze thawing 4 times again, centrifuges 15min under conditions of rotating speed is 8000r/min, supernatant and precipitation is separated, take and precipitate standby；
S2, to add volume fraction by the solid-to-liquid ratio 1g/mL of wet algae and ethanol into the precipitation described in step S1 be 90%
EtOH Sonicate is extracted 2 times, and the frequency of ultrasonic extraction is 40kHz, and temperature is 4 DEG C, and the time of ultrasound is 20min every time, in rotating speed
To centrifuge 15min under conditions of 8000r/min, supernatant and precipitation are separated, takes supernatant to be dried in vacuo, vacuum is done
Product after dry is dissolved with one-level water, and it is that (A represents that chlamydomonas is wet to 1gA/mL to adjust it relative to the concentration of chlamydomonas (wet algae) weight
Algae), produce chlamydomonas ethanol extract.
Influence of the chlamydomonas ethanol extract of embodiment 4 to Caenorhabditis elegans survival rate under oxidative stress conditions
Paraquat, scientific name methyl viologen is a kind of strong oxidizer, can induce a large amount of generation active oxygen radicals in vivo
(ROS) body is in oxidative stress environment, and then early ageing phenomenon occur.
Chlamydomonas ethanol extract made from Example 1, the wild type at adult initial stage is added to by final concentration 100mgA/mL
In Caenorhabditis elegans (N2) (being provided by Univ Minnesota-Twin Cities USA's Caenorhabditis elegans hereditary information collection), control group
Isometric S Medium solution is added, is placed in after 20 DEG C of culture 24h, the paraquat for adding final concentration of 50mM carries out the oxidation side of body
Compel modeling, the ratio of Caenorhabditis elegans survival is then counted every 12h, until it is all dead.Caenorhabditis elegans is deposited
Motility rate is represented with survivorship curve, is as a result analyzed with Kaplan-Meier methods.As a result it is as shown in Figure 1.
Fig. 1 results are shown, when chlamydomonas ethanol extract can extend the existence of Caenorhabditis elegans under oxidative stress conditions
Between, improve its survival rate.Result of the test shows that the chlamydomonas ethanol extract that the present invention is provided can be alleviated and caused by oxidative stress
Early ageing phenomenon, with significant anti-oxidant and anti-aging effects.
Influence of the chlamydomonas ethanol extract of embodiment 5 to Caenorhabditis elegans average life span under natural conditions
Final concentration of 100mgA/mL embodiment is added into the L4 phases Wild-type C. elegans (N2) after synchronization
The 1 chlamydomonas ethanol extract prepared, control group adds isometric S Medium solution, 20 DEG C of cultures is placed in, then every 1 day
The survival condition of Caenorhabditis elegans is counted, until it is all dead.The life-span of Caenorhabditis elegans is represented with survivorship curve, is tied
Fruit is analyzed with Kaplan-Meier methods.As a result it is as shown in Figure 2.
Fig. 2 results are shown, compared with control group, and chlamydomonas ethanol extract mutually can extend the average longevity of Caenorhabditis elegans
Life.Result of the test shows that the chlamydomonas ethanol extract that the present invention is provided has the effect of anti-aging process.
Influence of the chlamydomonas ethanol extract of embodiment 6 to ROS levels in Caenorhabditis elegans body
24 well culture plates are taken, if test group and control group, every group sets two holes, be 1mL per hole final volume, test group adds together
Chlamydomonas ethanol extract made from L4 phases Wild-type C. elegans (N2) and embodiment 1 after stepization, chlamydomonas ethanol is extracted
The final concentration 100mgA/mL of thing, control group add synchronize after L4 phases Wild-type C. elegans (N2) and with test group clothing
The isometric S Medium solution of algae ethanol extract, is placed in after 20 DEG C of culture 24h, and paraquat is added per hole to final concentration of
2mM, continues to cultivate after 48h, collects each group Caenorhabditis elegans, it is homogenized under condition of ice bath respectively, by homogenate 4
DEG C, 5min is centrifuged under the conditions of 10000g, collect supernatant, final concentration of 50 μM of DCFH-DA probes (purchase added into supernatant
In Sigma companies, No. CAS：2044-85-1) solution, using fluorescence microplate reader in 485nm excitation wavelengths and 538nm launch wavelengths
Lower fluorescence intensity, room temperature detection 2h, is surveyed once per 10min.As a result F check analyses are used, as a result as shown in Figure 3.
Fig. 3 results are shown, compared with control group, after being fed through chlamydomonas ethanol extract, ROS water in Caenorhabditis elegans body
It is flat to be remarkably decreased.Result of the test shows that the chlamydomonas ethanol extract that the present invention is provided can promote internal ROS removing, significantly drop
Low internal ROS levels, with significant oxidation resistance.
Above content is to combine specific preferred embodiment further description made for the present invention, it is impossible to assert
The specific implementation of the present invention is confined to these explanations.For general technical staff of the technical field of the invention,
On the premise of not departing from present inventive concept, some simple deduction or replace can also be made, should all be considered as belonging to the present invention's
Protection domain.
Claims (5)
1. a kind of chlamydomonas ethanol extract, it is characterised in that the preparation method of the chlamydomonas ethanol extract comprises the following steps：
S1, the wet algae of fresh chlamydomonas is taken, be to freeze 12 hours under conditions of -20 DEG C~-40 DEG C in temperature, it is small that room temperature melts 12
When, multigelation 3~5 times centrifuges 5~15min, by supernatant under conditions of rotating speed is 5000r/min~10000r/min
With precipitation separation, take precipitation standby；
S2, the EtOH Sonicate that addition volume fraction is 70%~99% into the precipitation described in step S1 are extracted 2~4 times, are being turned
Speed is 5~15min of centrifugation under conditions of 5000r/min~10000r/min, and supernatant and precipitation are separated, take supernatant to carry out
Vacuum drying, the one-level water of the product after vacuum drying is dissolved, chlamydomonas ethanol extract is produced.
2. chlamydomonas ethanol extract according to claim 1, it is characterised in that the preparation side of the chlamydomonas ethanol extract
Ethanol is added by the solid-to-liquid ratio 1g/mL of wet algae and ethanol carry out ultrasonic extraction in method, in step S2.
3. chlamydomonas ethanol extract according to claim 1, it is characterised in that the preparation side of the chlamydomonas ethanol extract
In method, the frequency of ultrasonic extraction is 35~40kHz in step S2, and temperature is 4 DEG C.
4. chlamydomonas ethanol extract according to claim 1, it is characterised in that the preparation side of the chlamydomonas ethanol extract
In method, the time of each ultrasonic extraction is 10~30min in step S2.
5. chlamydomonas ethanol extract according to claim 1 is being prepared with anti-oxidant and/or anti-senescence function health care
Application in food and medicine.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104001180A (en) * | 2014-05-16 | 2014-08-27 | 集美大学 | Preparation method of alga natural antioxidant |
| CN104274496A (en) * | 2013-07-10 | 2015-01-14 | 山东洁晶集团股份有限公司 | Extraction method for sargassum horneri polyphenol |
| CN105037300A (en) * | 2015-06-17 | 2015-11-11 | 宁波大学 | Method for comprehensively extracting fucoxanthin and brown alga polyphenol from gulfweed |
-
2017
- 2017-03-06 CN CN201710126725.9A patent/CN106974284A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104274496A (en) * | 2013-07-10 | 2015-01-14 | 山东洁晶集团股份有限公司 | Extraction method for sargassum horneri polyphenol |
| CN104001180A (en) * | 2014-05-16 | 2014-08-27 | 集美大学 | Preparation method of alga natural antioxidant |
| CN105037300A (en) * | 2015-06-17 | 2015-11-11 | 宁波大学 | Method for comprehensively extracting fucoxanthin and brown alga polyphenol from gulfweed |
Non-Patent Citations (2)
| Title |
|---|
| 赵淑江: "《海洋藻类生态学》", 31 May 2014, 海洋出版社 * |
| 邓祥元: "《应用微藻生物学》", 30 November 2016, 海洋出版社 * |
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