CN107019667A - The preparation method of artemia ovum hatching liquid extract - Google Patents
The preparation method of artemia ovum hatching liquid extract Download PDFInfo
- Publication number
- CN107019667A CN107019667A CN201710314772.6A CN201710314772A CN107019667A CN 107019667 A CN107019667 A CN 107019667A CN 201710314772 A CN201710314772 A CN 201710314772A CN 107019667 A CN107019667 A CN 107019667A
- Authority
- CN
- China
- Prior art keywords
- artemia
- hatching
- ovum
- preparation
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241001247197 Cephalocarida Species 0.000 title claims abstract description 35
- 230000012447 hatching Effects 0.000 title claims abstract description 35
- 102000002322 Egg Proteins Human genes 0.000 title claims abstract description 16
- 108010000912 Egg Proteins Proteins 0.000 title claims abstract description 16
- 210000004681 ovum Anatomy 0.000 title claims abstract description 16
- 239000007788 liquid Substances 0.000 title claims abstract description 7
- 238000002360 preparation method Methods 0.000 title claims abstract description 6
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims abstract description 5
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims abstract description 5
- 235000011130 ammonium sulphate Nutrition 0.000 claims abstract description 5
- 238000010612 desalination reaction Methods 0.000 claims abstract description 5
- 238000005119 centrifugation Methods 0.000 claims abstract description 3
- 238000005227 gel permeation chromatography Methods 0.000 claims abstract description 3
- 208000031513 cyst Diseases 0.000 claims description 10
- 238000001556 precipitation Methods 0.000 claims description 8
- 239000006228 supernatant Substances 0.000 claims description 8
- 239000002202 Polyethylene glycol Substances 0.000 claims description 5
- 229920001223 polyethylene glycol Polymers 0.000 claims description 5
- 239000013535 sea water Substances 0.000 claims description 4
- 238000002525 ultrasonication Methods 0.000 claims description 4
- 238000000926 separation method Methods 0.000 claims description 3
- 238000004108 freeze drying Methods 0.000 claims description 2
- 241000238582 Artemia Species 0.000 claims 1
- 239000008363 phosphate buffer Substances 0.000 claims 1
- 238000003756 stirring Methods 0.000 claims 1
- 238000000605 extraction Methods 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 12
- 238000000034 method Methods 0.000 description 4
- 210000001136 chorion Anatomy 0.000 description 3
- 238000009010 Bradford assay Methods 0.000 description 2
- 229920005654 Sephadex Polymers 0.000 description 2
- 239000012507 Sephadex™ Substances 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 210000004379 membrane Anatomy 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 150000003016 phosphoric acids Chemical class 0.000 description 2
- 241000143060 Americamysis bahia Species 0.000 description 1
- 241000238424 Crustacea Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000133262 Nauplius Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 239000008055 phosphate buffer solution Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/987—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K97/00—Accessories for angling
- A01K97/04—Containers for bait; Preparation of bait
- A01K97/045—Preparation of bait; Ingredients
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/10—General cosmetic use
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/82—Preparation or application process involves sonication or ultrasonication
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Dermatology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Cosmetics (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a kind of preparation method of artemia ovum hatching liquid extract.Whole hatching system is handled with ultrasonic wave after artemia ovum hatching, promotes the release of associated products, artemia hatching solution is collected by centrifugation and is precipitated with ammonium sulfate, be then freeze-dried after gel chromatography desalination.Extraction time is short, and loss of activity is small, reduces the introducing of foreign matter, and basis of reality is provided further to research and develop its function and purposes.
Description
Technical field
The present invention relates to a kind of extracting method of natural products, and in particular to the preparation side of artemia ovum hatching liquid extract
Method.
Background technology
Artemia is also referred to as shrimps, is a kind of crustaceans lived in high salt waters.Artemia contains abundant albumen
Matter, amino acid, unrighted acid and inorganic elements, are widely used as food organisms and experimental animal material.Artemia cysts is general
Refer to Artemia cysts, be mainly used to hatch artemia nauplii, as aquatic feed.Because artemia cysts is nutritious and is easy to protect
Deposit, can be with ready access upon use, its purposes also gradually expands, artemia cysts, chorion have started to be used for the conventional use such as cosmetics, food with
The active component included in outer field, especially hatching process in chorion, hatching membrane and artemia hatching solution also is being increasingly subject to close
Note.These compositions are after hatching later stage chorion and hatching membrane rupture, and part release is into artemia hatching solution, and content is extremely low, is comprising one
The enzyme relevant with hatching is arranged, activation and the hatching characteristic of ovum is directly affected.It is separated and extracted, for further research
Its function is significant with developing.
The content of the invention
It is an object of the invention to provide a kind of preparation method of artemia ovum hatching liquid extract, simple efficient separation is incubated
Change the active component in liquid.
To achieve the above object, the technical measures that the present invention takes are as follows：
1. after artemia cysts is fully cleaned, be hydrated, ovum is scattered in three in the ratio of every gram of dry ovum 50-100 milliliters of artificial seawater
In the bottle of angle, shaking table 100rpm/min vibrations, 25 DEG C of temperature；
2. after hatching 24-30 hours, 0.5%-1% polyethylene glycol, ultrasonication 20 seconds, power density are added in artemia hatching solution
0.1w-4w/cm2, centrifuge, collect artemia hatching solution.Precipitation is redissolved in 100 milliliters of artificial seawaters, stirred 5 minutes, centrifugation
Separation, centrifuged supernatant will merge twice；
3. artemia hatching solution is centrifuged, rotating speed 500g, 5 minutes, supernatant added ammonium sulfate, and precipitation is again molten with phosphate buffer
Solution, gel chromatography desalination, freeze-drying.
The invention has the advantages that：The higher-order of oscillation of the ultrasonic wave in artemia hatching solution effectively increases hatching membraned inclusions
Release, shortens extraction time, protects extract activity.Polyethylene glycol effectively protects nauplius from crushing simultaneously,
Reduce the introducing of impurity.Manually seawater all the components are, it is known that reduce influence of the natural sea impurities in water to extract.
Embodiment
The inventive method is described in further detail with reference to specific embodiment.
Embodiment 1
1. after artemia cysts is fully cleaned, be hydrated, ovum is scattered in triangular flask in the ratio of every gram of dry 80 milliliters of artificial seawater of ovum
In, shaking table 100rpm/min vibrations, 25 DEG C of temperature；
2. after hatching 24 hours, 0.5% polyethylene glycol, ultrasonication 20 seconds, power density 0.5w/cm are added in artemia hatching solution2,
Centrifuge, collect artemia hatching solution.Precipitation is redissolved in 100 milliliters of artificial seawaters, stirred 5 minutes, is centrifuged, will twice from
Supernatant merges；
3. artemia hatching solution is centrifuged, rotating speed 500g, 5 minutes, it was 50% that supernatant, which adds ammonium sulfate to saturation degree, by precipitation phosphoric acid
Salt buffer dissolves again, sephadex sephadexG-25 chromatography desalinations, freezes.
Determined through Bradford methods, protein content 53% in extract counts yield as 0.87% using dry artemia cysts.
Embodiment 2
1. artemia cysts is fully cleaned, ovum is scattered in triangular flask in the ratio of every gram of dry 100 milliliters of artificial seawaters of ovum, shaking table
100rpm/min vibrates, 25 DEG C of temperature；
2. after hatching 28 hours, 0.8% polyethylene glycol, ultrasonication 20 seconds, power density 0.5w/cm are added in artemia hatching solution2,
Centrifuge, collect artemia hatching solution.Precipitation is redissolved in 100 milliliters of artificial seawaters, stirred 5 minutes, is centrifuged, will twice from
Supernatant merges；
3. artemia hatching solution is centrifuged, rotating speed 500g, 5 minutes, it was 55% that supernatant, which adds ammonium sulfate to saturation degree, by precipitation phosphoric acid
Salt buffer dissolves again, sephadex sephadexG-25 chromatography desalinations, freezes.
Determined through Bradford methods, protein content 55% in extract counts yield as 0.94% using dry artemia cysts.
Claims (1)
1. the preparation method of artemia ovum hatching liquid extract, it is characterized in that step is as follows：
（1）After artemia cysts is fully cleaned, being hydrated, ovum is scattered in three in the ratio of every gram of dry ovum 50-100 milliliters of artificial seawater
In the bottle of angle, shaking table 100rpm/min vibrations, 25 DEG C of temperature；
（2）After hatching 24-30 hours, 0.5%-1% polyethylene glycol, ultrasonication 20 seconds, power density are added in artemia hatching solution
0.1w-4w/cm2, centrifuge, collect artemia hatching solution, precipitation is redissolved in 100 milliliters of artificial seawaters, stir 5 minutes, centrifugation
Separation, centrifuged supernatant will merge twice；
（3）Artemia hatching solution is centrifuged, rotating speed 500g, 5 minutes, supernatant adds ammonium sulfate, by precipitation with phosphate buffer again
Dissolving, gel chromatography desalination, freeze-drying.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710314772.6A CN107019667A (en) | 2017-05-07 | 2017-05-07 | The preparation method of artemia ovum hatching liquid extract |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710314772.6A CN107019667A (en) | 2017-05-07 | 2017-05-07 | The preparation method of artemia ovum hatching liquid extract |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107019667A true CN107019667A (en) | 2017-08-08 |
Family
ID=59530221
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710314772.6A Pending CN107019667A (en) | 2017-05-07 | 2017-05-07 | The preparation method of artemia ovum hatching liquid extract |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107019667A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1699561A (en) * | 2005-05-05 | 2005-11-23 | 中国海洋大学 | Process for preparing artemia hatching enzyme by using artemia hatching liquid |
| CN104273099A (en) * | 2014-10-21 | 2015-01-14 | 天津海友佳音生物科技股份有限公司 | Method for increasing eclosion rate of diapause artermia eggs at high temperature |
| CN104285911A (en) * | 2014-10-21 | 2015-01-21 | 天津海友佳音生物科技股份有限公司 | Method for hatching diapause brine shrimp eggs under blue light |
-
2017
- 2017-05-07 CN CN201710314772.6A patent/CN107019667A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1699561A (en) * | 2005-05-05 | 2005-11-23 | 中国海洋大学 | Process for preparing artemia hatching enzyme by using artemia hatching liquid |
| CN104273099A (en) * | 2014-10-21 | 2015-01-14 | 天津海友佳音生物科技股份有限公司 | Method for increasing eclosion rate of diapause artermia eggs at high temperature |
| CN104285911A (en) * | 2014-10-21 | 2015-01-21 | 天津海友佳音生物科技股份有限公司 | Method for hatching diapause brine shrimp eggs under blue light |
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| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170808 |