CN103509047B - The extraction process of the phosphatidylcholine of a kind of antarctic krill and the preparation method of Phosphatidylserine - Google Patents
The extraction process of the phosphatidylcholine of a kind of antarctic krill and the preparation method of Phosphatidylserine Download PDFInfo
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Abstract
The present invention provides the extraction process of the phosphatidylcholine of a kind of antarctic krill, comprise the steps: to collect Antarctic krill head, boiling, stirring, krill head decoction liquor is delivered to solid-liquid separation in cold pressing expeller, it is thus achieved that rich in oily liquids and the first-class solid of krill of phospholipid；Adding dehydrated alcohol in the first-class solid of krill, high shear extracts, and centrifugation, concentration obtain oily liquids；Merge oily liquids, drying under reduced pressure, then through flame filter press filter remove insoluble substance, after through membrane separation concentration, column chromatographic isolation and purification, lyophilization, it is thus achieved that product phosphatidylcholines.Present invention also offers the preparation method of the Phosphatidylserine of antarctic krill, the phosphatidylcholine extracted hydrolyzes through Choline phosphatase, extraction, it is thus achieved that rich in the Phosphatidylserine of DHA and EPA.Extraction process of the present invention and preparation method environmental protection, low cost, it is thus achieved that product content high, activity is good, can be with industrialized production.
Description
Technical field
The present invention relates to a kind of deep working method from Antarctic krill, be specifically related to from Antarctic krill extract phosphatidylcholine and
The method preparing Phosphatidylserine.
Background technology
Phosphatidylserine (PS) rich in unsaturated fatty acid docosahexenoic acid (DHA) prevents senile dementia as future,
Brain health such as preventing children hyperkinetic syndrome and prevent cardiovascular emerging health care and medical material to have wide market prospect.
Antarctic krill is the one way of life krill in waters, the Antarctic Continent, is also that on the earth, quantity is maximum, it is raw to multiply most successful single
One of goods and materials source.The krill oil extracted from Antarctic krill is rich in functional components such as phospholipid (mainly based on lecithin), and phosphorus
The fatty acid of fat forms based on eicosapentaenoic acid (EPA) and docosahexenoic acid (DHA), and EPA and DHA content account for
More than the 40% of fatty acids in phospholipids total amount.Antarctic krill oil has reduction cardiovascular disease, prevention senile dementia and improves body
The multiple physiologically active such as immunity, and its activity is better than triglyceride type EPA/DHA product in the market and terrestrial plant
The lecithin product (without EPA and DHA in its fatty acid composition) in source, unsaturated fatty acid docosahexenoic acid (DHA)
It is mainly in combination with on glyceryl 2 of phosphatidyl choline (Sn2 position), the unsaturated fatty acid 20 that namely we are previously mentioned
The phosphatidyl choline PC that two carbon acids (DHA) combine.DHA is to constitute cephalin, the basis of brain cell membrane, divides brain cell
Split, breed, nerve conduction, the g and D of synapse play particularly important effect, are that human brain is formed and IQ is developed
Required material.It is to vision, cerebral activity, lipid metabolism, fetal growth and immunologic function and avoids alzheimer disease all
There is extreme influence, series of symptoms during shortage, can be caused, including growth retardation, skin abnormality squama, sterile, intelligence barrier
Hinder.But common DHA is to arrange in a random way, is in unordered state, it is impossible to guarantee to be absorbed by the body completely,
The generally DHA on Sn1, Sn3 position can be burned generation heat, or combines the fatty acid on Sn2, forms triglyceride,
DHA on only Sn2 position could be fully absorbed by human body.Krill oil is DHA on the Sn2 position of phospholipid, it is possible to completely by people
Absorbing, according to our data research, the biological activity of the DHA on Sn2 and bioavailability are common fish oil DHA activity
3-5 times.Simultaneously by krill rich in the phosphatidyl choline way by biological enzyme, change into Phosphatidylserine.Phosphatidyl
Its function of serine mainly improves neurocyte function, the conduction of regulation Nerve impulse, promotes brain memory function.Therefore warp
Cross studying for a long period of time and summing up of inventor, the phosphorus combined from the unsaturated fatty acid docosahexenoic acid (DHA) in krill oil source
Acyl serine, due to its comprehensive function mechanism, for improving brain neuroblastoma cell function and cerebral activity, improves memory and rush
Enter infant brain development and safeguard that brain health has the problem meaning of great economic worth and biological medicine research.
Under constantly the increasing of krill deep processing, how to improve and krill processing is extracted in garbage phospholipid and deep processing krill oil
Phospholipid, and then to change into Phosphatidylserine be then that current biological medicine gets important focus.The most universal technology solution party
Case has three kinds:
The first is directly to purchase the most existing krill oil finished product, carries out the further separating-purifying of phosphatidylcholine.Patent
CN102405988A, applies for that artificial Liaoning Dalian Ocean Fishery Group Corporations, patent name " extract high phospholipid to contain from krill
The method of amount krill oil "；Patent CN102439125A, obligee Shanghai Ying Bei Bioisystech Co., Ltd, patent of invention "
The method planting the high quality Antarctic krill oil preparing the Phosphatidylserine rich in many unsaturated double-bonds fatty acyl group " and Li Bogen
(Lipogen) pure Superba krill oil (the Superba Krill of the biotech company Aker BioMarine of Norway is extracted from
Oil), form krill oil to originate 10% Phosphatidylserine product.At present the price of this raw material itself the most 200 U.S. dollars with
Upper per kilogram, is about more than 300-5000 U.S. dollar through the product cost converted, owing to this product exists complex manufacturing and numerous
Trivial, need removing protein and ionized impurity could form mobility final product as oil krill oil, especially various countries are at present for food
The regulation of raw material is increasingly stricter, is eventually fabricated satisfactory cost of material high, and the unsaturated lipid rich in phospholipid of oily
Fat acid forms the extraction separation of the liquid configuration before Phosphatidylserine and product for rear road after catalysis, increases a lot of difficulty,
Causing product content on the low side, free fatty acid content height, poor fluidity, muddiness and dissolvent residual such as exceed standard at the defect, do not obtain
Take the optimal industrialization way of industrialized production Phosphatidylserine.
The second process route is to use organic solvent or multiple organic solvent to extract, at present about the similar technique extracted
Such as Neptune company of Canada is the extracting method (patent No. EP01417211B1) of representative, adds ethanol and acetic acid with acetone
The mixed solvents such as ethyl ester carry out magazins' layout and enrichment concentrates.But it is as the further of our existing environmental protection and Safety production questions
Strictly regulating, managing, for sewage disposal and safety in production, the cost controlled can be more and more higher, including similar technique environmental impact assessment and
The biggest difficulty is just had on approving and initiate a project.And existing much include that a lot of solvent extractions are all proposed by Japan, European Union and China
A lot of laws and regulations requirements, such as acetone cannot function as food to be added and the extractant of health promoting product, thus similar products are in the future
The biggest regulation obstacle is had on entering part international market.Again, owing to using organic solvent, particularly portioned product to include first
Alcohol and acetone etc., will remove residual to the later stage increases a lot of operations and difficulty.
The third process route is to use supercritical extraction.CO 2 supercritical is all that comparison is feasible for test and theory stage
Means, but really put into a set of equipment and need the input of the most more than one hundred million RMB could really form industrialization.According to this
The research of a person of good sense, this process route, there is the biggest equipment investment requirement, even if going into operation, due to the problem of equipment depreciation, with
Sample is also high cost, and carbon dioxide supercritical fluid extraction is a kind of extensive extraction means simultaneously, it is impossible to the required phospholipid of enrichment targetedly
The unsaturated fatty acid that content is high, thus also have its limitation.This process route refers to CN102358865A, Shandong section Rui
" a kind of method of extracting Euphausia superba oil by using supercritical carbon dioxide " of your biological product company limited application etc..
Therefore, this area need to develop the extraction process of the phosphatidylcholine of new environmental protection, lower-cost antarctic krill and
The preparation method of Phosphatidylserine.
Summary of the invention
It is an object of the invention to provide the extraction process of the phosphatidylcholine (PC) of a kind of environmental protection, lower-cost antarctic krill.
Second object of the present invention also provides for the preparation method of the Phosphatidylserine (PS) of a kind of economy, environmental protection.
A first aspect of the present invention, it is provided that the extraction process of the phosphatidylcholine of a kind of antarctic krill, described extraction process bag
Include following step:
A () collects Antarctic krill head, add water, and decocts, and krill head decoction liquor is delivered to solid-liquid separation in cold pressing expeller, obtained by stirring
Must be rich in the first-class solid of the oily liquids of phospholipid and krill；
B () adds dehydrated alcohol in the first-class solid of krill, high shear extracts, and centrifugation obtains supernatant；This step carry out
1-2 time, merge supernatant, concentrate and obtain oily liquids；
C oily liquids in step (a) and step (b) is merged by (), drying under reduced pressure, it is thus achieved that containing phospholipid and unsaturated lipid
The protein coagulation thing of fat acid；
D protein coagulation thing containing phospholipid and unsaturated fatty acid is dissolved in food stage organic solvent by (), through flame filter press mistake
Filtering off except insoluble substance, the liquid obtained after filtration is again through membrane separation concentration, it is thus achieved that concentrated solution；
E () concentrated solution is through column chromatographic isolation and purification, it is thus achieved that phosphatidylcholine mixture；The most freeze-dried, it is thus achieved that phosphatidyl gallbladder
Alkali product.
In above-mentioned steps (a) in krill head 2h after thawing, add water, 40-55 DEG C of decoction, add ethanol during stirring.Described second
The percentage by weight of alcohol is 5%.
In above-mentioned steps (b), supernatant is through concentrating under reduced pressure, it is thus achieved that oily liquids.
In above-mentioned steps (d), food stage organic solvent is ethanol and normal hexane.
Ethanol in above-mentioned steps (d): the volume ratio of normal hexane is 2-4:0.5-1.5.Preferred alcohol: the volume ratio of normal hexane is
3:1。
In above-mentioned steps (d), using Ultra filtration membrane to concentrate, solid-liquid ratio W:V is 1:20, temperature 10-55 DEG C, uses bore
100nm → 20nm bi-membrane method separates.
In above-mentioned steps (d), using micro-filtration membrane or ultrafilter membrane in membrance separation, the aperture of described film is 0.02-10 μm, at film
Upper applying pressure 0.015 ~ 10MPa.
Concentrated solution is first dissolved in by above-mentioned steps (e) in the flowing mutually of normal hexane, isopropanol, water composition, fixes and select mutually
50-400 mesh silochrom or macroporous resin.
During above-mentioned steps (e) column chromatography, described pillar blade diameter length ratio 1:5-10, described eluant selected from normal hexane aqueous solution,
The aqueous solution of isopropanol or normal hexane and the aqueous solution of isopropanol.
Above-mentioned fixing selection 100-200 mesh silochrom mutually.
A second aspect of the present invention, it is provided that the preparation method of the Phosphatidylserine of a kind of antarctic krill, described preparation method
Comprise the steps:
A () collects Antarctic krill head, boiling, stirring, krill head decoction liquor is delivered to solid-liquid separation in cold pressing expeller, it is thus achieved that
Oily liquids and the first-class solid of krill rich in phospholipid；
B () adds dehydrated alcohol in the first-class solid of krill, high shear extracts, and centrifugation obtains supernatant；This step extracts
1-2 time, merge supernatant, concentrate and obtain oily liquids；
C oily liquids in step (a) and step (b) is merged by (), drying under reduced pressure, it is thus achieved that containing phospholipid and unsaturated lipid
The protein coagulation thing of fat acid；
D protein coagulation thing containing phospholipid and unsaturated fatty acid is dissolved in food stage organic solvent by (), through flame filter press mistake
Filtering off except insoluble substance, the liquid obtained after filtration is again through membrane separation concentration, it is thus achieved that concentrated solution；
E () concentrated solution is through column chromatographic isolation and purification, it is thus achieved that phosphatidylcholine mixture；The most freeze-dried, it is thus achieved that phosphatidyl gallbladder
Alkali；
F (), in aqueous phase, adds Serine, Choline phosphatase, calcium chloride, hydrolytic phosphatide phatidylcholine, generates phosphatidyl silk ammonia
Acid；Remove solvent, it is thus achieved that the thick product of Phosphatidylserine；
G thick for Phosphatidylserine product is repeatedly extracted by () through 2-4 food stage organic solvent, adjusted by pH for the first time during extraction
Joint, to 3, removes organic solvent, it is thus achieved that Phosphatidylserine product.
In above-mentioned steps (f), phosphatidylcholine is in aqueous phase during hydrolysis, and temperature is 40-45 DEG C, pH5-6, described phosphorus
Phosphatidylcholine: Serine: Choline phosphatase is 1:7-13:0.03-0.06.
In above-mentioned steps (f), below 50 DEG C, decompression method is used to remove solvent.
In above-mentioned steps (g), for the first time during extraction, the mixture that food stage organic solvent is ethanol and normal hexane of use, institute
State ethanol and normal hexane volume ratio is 4:1, and use edible salt acid for adjusting pH to 3, remove upper liquid.
In above-mentioned steps (g), using normal hexane to carry out follow-up 1-3 extraction, extraction terminates to remove upper liquid every time.
In above-mentioned steps (g), repeatedly after extraction, use 90% ethanol water, remove normal hexane, obtain Phosphatidylserine
Product.
In above-mentioned steps (d), food stage organic solvent is ethanol and normal hexane.
In above-mentioned steps (d), ethanol: the volume ratio of normal hexane is 2-4:0.5-1.5.Preferred alcohol: the volume ratio of normal hexane
For 3:1.
In above-mentioned steps (a) in krill head 2h after thawing, add water, 40-55 DEG C of decoction, add ethanol during stirring.Described second
The percentage by weight of alcohol is 5%.
In above-mentioned steps (b), supernatant is through concentrating under reduced pressure, it is thus achieved that oily liquids.
In above-mentioned steps (d), using Ultra filtration membrane to concentrate, solid-liquid ratio W:V is 1:20, temperature 10-55 DEG C, uses bore
100nm → 20nm bi-membrane method separates.
In above-mentioned steps (d), using micro-filtration membrane or ultrafilter membrane in membrance separation, the aperture of described film is 0.02-10 μm, at film
Upper applying pressure 0.015 ~ 10MPa.
Concentrated solution is first dissolved in by above-mentioned steps (e) in the flowing mutually of normal hexane, isopropanol, water composition, fixes and select mutually
50-400 mesh silochrom or macroporous resin.
During above-mentioned steps (e) column chromatography, described pillar blade diameter length ratio 1:5-10, described eluant selected from normal hexane aqueous solution,
The aqueous solution of isopropanol or normal hexane and the aqueous solution of isopropanol.
Above-mentioned fixing selection 100-200 mesh silochrom mutually.
A third aspect of the present invention, it is provided that according to the phosphatidylcholine of the antarctic krill that said extracted technique is extracted.
A fourth aspect of the present invention, it is provided that according to the Phosphatidylserine of antarctic krill prepared by above-mentioned preparation method.
" cleaning in krill head 2h after thawing " of the present invention refers to: krill head is the most freezing, extracts phosphatidyl
During choline, boiling in krill head 2h after thawing；Or, after freezing krill is thawed in 2h, collect krill head,
And carry out boiling.
Advantages of the present invention and effect be:
1. the krill head of Aquatic product company is particularly concentrated decoction, cold pressing, high shear extraction, plate-and-frame filtration, film in time by the present invention
The pre-treatments such as separation, column chromatography, according to the characteristic of phospholipid prod, specific aim chooses multiple physics and the combination of nanometer extraction process,
The oils and fats etc. using membrane ultrafiltration process filtration will not be polymerized in normal hexane in production technology, greatly reduces the purification of subsequent product
Load with separating, reduces the impurity upper prop of follow-up resin absorption, improves absorption equivalent, improve the yield of product, enter
One step improves quality and the stability of product, has economically saved production cost simultaneously, and has greatly reduced soda acid and second
The consumption of alcohol.And raw material is also used as other purposes from Aquatic product company garbage, the by-product formed in extraction process, tool
There is the social economic effect of maximum.From the point of view of from environmental protection, greatly reduce the discharge of waste water, alleviate environmental protection pressure；From production
On say, whole system is easily maintained, and substantially improves operating environment, reduces labor intensity.
2. present invention process compares the most universal solvent-extracted separating-purifying means, improves the yield of PC, changed
Going an actual industrial difficult problem for the low yield of PC preparation process and waste water high pollution, the industrialization simultaneously comparing CO 2 supercritical is thrown
Money, equipment investment only have 3% less than.This technique effectively prevent effective ingredient activity in high temperature and batch technology in Antarctic krill
The oxidation of composition and degraded so that the industrialized production of the refined DHA-PS product obtaining high-load is accomplished.
The feature of this technique is first to prepare the Phosphatidylserine of high-load unsaturated fatty acid docosahexenoic acid (DHA)
(PS) product is final purpose, is extracted the specific aim compound mode of means, the extraction phosphorus that energy is economic and environment-friendly by temperature and physics
Phospholipid in the waste phosphorus prawn head of shrimp processing factory, and by cold pressing, high shear extracts, membrane filtration, the method such as high speed centrifugation and chromatography
Comprehensive use, go the removal of impurity by screening and separating targetedly so that unsaturated fatty acid 22 carbon six in mix products
The content of olefin(e) acid (DHA) phospholipid can reach more than 45%.
Below in conjunction with embodiment, the present invention is described in detail.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described further, and its specific embodiments is construed as being only for example
Bright, it not determinate, it is impossible to limit protection scope of the present invention with following illustration.
Inventive concept is: for weak point present in the real industrialization of prior art provide a kind of by Antarctic krill head and
Shi Jizhong pre-treating technology, and effectively prevent effective ingredient oxidation of active component in high temperature and batch technology in Antarctic krill
And degraded, extract to greatest extent and separate high-content phosphorus phosphatidylcholine so that obtain the krill source Phosphatidylserine of high-load
Industrialized production can be accomplished.
Experimental technique
Annual late July starts Antarctic krill and fishes for period, carries out the first-class resource of krill that aquatic products processing factory is discarded unifying to collect,
Obtain about raw material batch 2000kg, ventilate room temperature 15 DEG C and thaw.3 batch samples have been carried out extracting separation by the present invention
Experiment, from example 1 below～3, experimental data understands: produce the high phospholipid processing technique in the antarctic krill oil source of DHA-PS
Being feasible, its Phosphatidylserine PS product rich in DHA ultimately formed is content more than 45%, detects two in product
Class active substance, wherein DHA content is up to more than 15%, and PS is up to more than 45%.It has DHA and PS two class activity simultaneously
Function, can as produce natural health and medicine raw material, this raw material with by the mixing of these two kinds of one matters of DHA and PS
Thing is compared, and biological activity improves 3-5 times, is the raw material of the great potential of production and processing food, medicine and field of health care food.
In order to extract separation high-content phosphorus phosphatidylcholine to greatest extent, preferably boiling pretreatment in 2h after the defrosting of krill head；Can also
Directly after aquatic products processing factory krill is thawed, the krill head boiling pretreatment will collected in 2h, then carry out follow-up extraction work
Skill.
The preparation 1 of embodiment 1 Phosphatidylserine
(1) by first-class for the 2000kg Antarctic krill alcohol extraction decoction pot putting 5 tons, pour 2000L water, heat 50 DEG C, for follow-up
Cold pressing is convenient, sprays 5% edible ethanol while stirring, and edible ethanol percentage by weight can be 3-6%, and general 5% is optimal,
The lubrication of squeezing can be increased.Stirring 10min, is then delivered to cold pressing expeller (model 6YL-165 type, river krill head decoction liquor
South chats) in solid-liquid separation, cold pressing expeller treating capacity 1000kg/h, rotating speed 28-35r/min, power 30kw, it is thus achieved that rich in phosphorus
The oily liquids of fat and the first-class solid of krill；
(2) solid that (1) obtains is placed in reactor, adds the anhydrous ethanol solvent of 4 times of gained solids of quality, high
Shear agitation 20min(15 ~ 30min), make phospholipid fraction be substantially dissolved in centrifugation in ethanol；Precipitate is put into
Reactor adds the anhydrous ethanol solvent high shear stirring of 4 times of gained solids of quality, centrifugal, twice supernatant decompression of gained
It is condensed into oily liquids；
(3) with (2) gained, (1) gained oily liquids being concentrated oily liquids to merge, decompression rotation is evaporated, and obtains containing phosphorus
The protein coagulation thing 589kg of fat and unsaturated fatty acid.
(4) in the protein coagulation thing containing phospholipid and unsaturated fatty acid that our detection (3) obtains, moisture is gross weight
67%, take a small amount of protein coagulation thing, its low-temperature evaporation is dried, detect dried protein coagulation thing, referred to as follows
Mark
| Composition | Account for the percentage ratio (%) of dry weight after drying |
| TL | 43.9 |
| Crude protein | 47.5 |
| Ash | 8.6 |
Above testing result tentatively shows, phospholipid and unsaturated fatty acid from krill head source are effective by being concentrated to give
Separating-purifying.Further by the composition of fatty acid is analyzed, Agilent company of U.S. gas chromatogram 6890N is used to divide
Analysis, the analytical column used is the 122-7032 of J&S Scientific DB-Wax.Analyze gained and contain phospholipid and unsaturated lipid
In the protein coagulation thing of fat acid, total fatty acid content is 63.5%, insatiable hunger during wherein unsaturated fatty acid content is the 35.5%(present invention
It mostly is Omega-3 series, wherein DHA7.8%, EPA content 15.0% with fatty acid.
(5) the protein coagulation thing containing phospholipid and unsaturated fatty acid obtained in (3) is placed in 5 tons of dissolving tanks, configuration
3000L food stage organic solvent, wherein ethanol-normal hexane (75:25 volume ratio), use stainless steel multilayer plate and frame filter pressing,
Multiple filtration plate can be used according to practical condition, the present embodiment uses the standard type of 8 sheet frames, by pressure
Contracting sheet frame, carries out solid and the separation of liquid and purification.By plate-and-frame filtration, remove insoluble substance, then to isolated
Liquid carries out Ultra filtration membrane, deoil (removing at the oils and fats not being polymerized in normal hexane), solid-liquid ratio 1:20(w/v), film
Pressure 0.1Mpa, uses bore 100nm → 20nm bi-membrane method to separate.Prior to 25 DEG C, 100nm membrane filtration, backflow and infiltration
Liquid crosses 20nm film in 25 DEG C, and backflow and penetrating fluid volume ratio reach 1:3, collect 20nm film backflow, obtain unsaturated fatty acids
The concentrated solution of acid, takes a small amount of concentrated solution evaporation drying, the protein coagulation thing containing phospholipid and unsaturated fatty acid after detection concentration
In each constituent content.
| Composition | Account for the percentage ratio (%) of dry weight after drying |
| TL | 52.5 |
| Crude protein | 40.4 |
| Ash | 7.1 |
Analyzing total fatty acid content in the protein coagulation thing that gained contains phospholipid and unsaturated fatty acid is 63.5%, wherein unsaturated
Content of fatty acid is 35.5%(DHA7.8%, EPA content 15.0%).
(6) by concentrated solution flowing phase normal hexane: isopropanol: (this ratio is in water (1:1:0.16, v/v/v) dissolving
Good ratio), stir to dissolving standby under room temperature.Learn from else's experience activation No. 2 silica gel of gross porosity, add flowing makes pulpous state, wet method mutually
Dress post, then balances each other chromatographic column (model Φ 315mm*2000mm) with flowing.The sample prepared slowly is poured in post,
Flowing phase eluting is added after sample introduction.Controlling void tower flow velocity is 0.382cm/min, and sample introduction concentration is 0.25g concentrated phosphatide (mL/
Flowing phase) room temperature condition under, fixing can select 50-400 mesh silochrom or macroporous resin mutually, the present embodiment is fixed phase
Selecting No. 2 silica gel of 100 ~ 200 mesh gross porosity, purification rate is 70%, and the suitableeest upper carrying capacity is 0.086gPC(g/ silica gel), through post layer
Analysing isolated and purified, it is thus achieved that eluent, obtaining PC content is more than 80%.Eluent can use the aqueous solution of normal hexane, different
The aqueous solution of propanol or normal hexane and the aqueous solution of isopropanol.Employing normal hexane in the present embodiment: isopropanol: water (1:1:0.16,
V/v/v) above-mentioned eluent (the i.e. phosphatidylcholine mixture) warp after membrane separation concentration and column chromatographic isolation and purification of eluent
Lyophilization, can obtain product phosphatidylcholines.
(7) in aqueous phase, add Serine, calcium chloride and Choline phosphatase, hydrolytic phosphatide phatidylcholine, generate phosphatidyl silk ammonia
Acid, during hydrolysis, temperature is 40-45 DEG C, pH5-6.In the present embodiment, in 3000L fermentation tank, put into (6) gained
Product phosphatidylcholines 50kg, Serine 500kg, calcium chloride 4.2kg reaction, add water to 500L, then put into phospholipid
Enzyme D (PLD enzyme) 1.0kg, after reacting 4h, then puts into the PLD enzyme of 0.5kg under the conditions of bath temperature 40-45 DEG C, pH5.6
Reaction 12h.The phosphatidylcholine race (PC) rich in many unsaturated double-bonds fatty acyl group (4) obtained, at aqueous phase reactions body
System is converted into the Phosphatidylserine (PS) rich in many unsaturated double-bonds fatty acyl group, the mixture after reaction 50 DEG C with
Solvent is removed in lower decompression, it is thus achieved that high-load Phosphatidylserine crude product 46.9kg, Phosphatidylserine therein (PS) content
It is about 39.8%.
(8) in the 46.9kg product (7) obtained, input 8000L, with in upper container, adds 1600L normal hexane and ethanol
400L, is subsequently adding the edible hydrochloric acid of 10-20L, is adjusted to pH3, is sufficiently stirred for, is layered, and removes upper liquid, at subnatant
In add 2000L normal hexane, stir, be layered, remove upper liquid be repeated 3 times.Add water in the solution: ethanol (1:9,
V/v) 150-300L, is thoroughly mixed, and regulates below pH9, separates hexane solution, removes the phosphatidyl silk after normal hexane
Propylhomoserin product carries out efficient liquid phase chromatographic analysis, and the content recording Phosphatidylserine (PS) is 45.9%.
The preparation 2 of embodiment 2 Phosphatidylserine
(1) by first-class for the 2050kg Antarctic krill alcohol extraction decoction pot putting 5 tons, pour 2000L water, heat 55 DEG C, for follow-up
Cold pressing is convenient, sprays 6% ethanol while stirring a small amount of, stirs 10min, then krill head decoction liquor is delivered to cold pressing expeller (type
Number 6YL-165 type, Henan chats) in solid-liquid separation, cold pressing expeller treating capacity 1000kg/h, rotating speed 28-35r/min, power
30kw, it is thus achieved that rich in oily liquids and the first-class solid of krill of phospholipid；
(2) solid that (1) obtains is placed in reactor, adds the anhydrous ethanol solvent of 3 times of gained solids of quality, high
Shear agitation 15min, makes phospholipid fraction be substantially dissolved in centrifugation in ethanol；Precipitate is put into reactor and adds quality 3
The anhydrous ethanol solvent high shear stirring of times gained solid, centrifugal, twice supernatant concentrating under reduced pressure of gained becomes oily liquids；
(3) with (2) gained, (1) gained oily liquids being concentrated oily liquids to merge, decompression rotation is evaporated, and obtains containing phosphorus
The protein coagulation thing 608kg of fat and unsaturated fatty acid.
(4) with embodiment 1, moisture in the protein coagulation thing containing phospholipid and unsaturated fatty acid that detection (3) obtains
For the 62% of gross weight, take a small amount of protein coagulation thing, its low-temperature evaporation is dried, detect dried protein coagulation thing,
To following index
| Composition | Account for the percentage ratio (%) of dry weight after drying |
| TL | 44.9 |
| Crude protein | 48.2 |
| Ash | 6.9 |
Further by the composition of fatty acid is analyzed, Agilent company of U.S. gas chromatogram 6890N is used to analyze, institute
The 122-7032 that analytical column is J&S Scientific DB-Wax used.Analyzing total fatty acid content in gained coagulum is 62%,
Wherein unsaturated fatty acid content is 37%(DHA8.1%, EPA content 15.6%).
(5) product is placed in 5 tons of dissolving tanks, configures 3000L food stage organic solvent, wherein ethanol-normal hexane (8:3),
By plate-and-frame filtration, remove insoluble substance etc., and then liquid to isolated carries out Ultra filtration membrane, deoils, feed liquid
Than 1:20(w/v), film pressure 0.1Mpa, use bore 100nm → 20nm bi-membrane method to separate.Prior to 10 DEG C, 100nm film
Filtering, backflow crosses 20nm film with penetrating fluid in 10 DEG C, and backflow and penetrating fluid volume ratio reach 1:3, collects the backflow of 20nm film
Liquid, obtains the concentrated solution of unsaturated fatty acid.
| Composition | Account for the percentage ratio (%) of dry weight after drying |
| TL | 52.9 |
| Crude protein | 40.6 |
| Ash | 6.5 |
Analyzing total fatty acid content in gained protein coagulation thing is 62%, and unsaturated fatty acid content is 37%(wherein DHA8.1%,
EPA content 15.6%).
(6) by concentrated solution flowing phase normal hexane: isopropanol: water (1:1:0.16, v/v/v) dissolves, stir under room temperature
Standby to dissolving.Learn from else's experience No. 2 silica gel of gross porosity of activation, add flowing and makes pulpous state mutually, wet method dress post, then equal with flowing
Weighing apparatus chromatographic column.The sample prepared slowly is poured in post, after sample introduction, adds flowing phase eluting.Controlling void tower flow velocity is
0.382cm/min, sample introduction concentration is under the room temperature condition of 0.25g concentrated phosphatide (mL/ flow phase), fixing selects 100 ~ 200 mutually
No. 2 silica gel of mesh gross porosity, purification rate is 70%, and the suitableeest upper carrying capacity is 0.086gPC(g/ silica gel), obtain PC content be 80% with
On phosphatidylcholine mixture.By phosphatidylcholine mixture lyophilization, it is thus achieved that product phosphatidylcholines.
(7) in aqueous phase, add Serine, calcium chloride and Choline phosphatase, hydrolytic phosphatide phatidylcholine product, generate phosphatidyl
Serine, during hydrolysis, temperature is 40-45 DEG C, pH5-6.In the present embodiment, in 3000L fermentation tank, put into (6)
The phosphatidylcholine mixture 50kg of gained, Serine 650kg, calcium chloride 20kg reaction, add water to 500L, then put into phosphorus
ESD (PLD enzyme) 3kg, at bath temperature 40-45 DEG C, pH5-6, after the present embodiment is adjusted to pH5.8 reaction 4h, then
Mixture after putting into the PLD enzyme reaction 12h reaction of 0.8kg reduces pressure below 50 DEG C and removes solvent, it is thus achieved that rich in many insatiable hungers
With Phosphatidylserine (PS) the crude product 46.3kg of double bond fatty acyl group, POPS therein (PS) content is about 39.1%.
(8) in the 46.3kg product (7) obtained, input 8000L, with in upper container, adds 1600L normal hexane and ethanol
400L, is subsequently adding the edible hydrochloric acid of 10-20L, is adjusted to pH3, is sufficiently stirred for, is layered, and removes upper liquid, at subnatant
In add 2000L normal hexane, stir, be layered, be repeated 3 times.Add water in the solution: ethanol (1:9, v/v) 150-300L,
Being thoroughly mixed, regulate below pH9, separate hexane solution, the product removing normal hexane carries out efficient liquid phase chromatographic analysis,
The content recording POPS (PS) is 45.0%.
The preparation 3 of embodiment 3 Phosphatidylserine
(1) by first-class for the 2010kg Antarctic krill alcohol extraction decoction pot putting 5 tons, pour 2000L water, heat 45 DEG C, for follow-up
Cold pressing is convenient, sprays 3% ethanol while stirring a small amount of, stirs 10min, then krill head decoction liquor is delivered to cold pressing expeller (type
Number 6YL-165 type, Henan chats) in solid-liquid separation, cold pressing expeller treating capacity 1000kg/h, rotating speed 28-35r/min, power
30kw, it is thus achieved that rich in oily liquids and the first-class solid of krill of phospholipid；
(2) solid that (1) obtains is placed in reactor, adds the anhydrous ethanol solvent of 3 times of gained solids of quality, high
Shear agitation 15min, makes phospholipid fraction be substantially dissolved in centrifugation in ethanol；Precipitate is put into reactor and adds quality 3
The anhydrous ethanol solvent high shear stirring of times gained solid, centrifugal, twice supernatant concentrating under reduced pressure of gained becomes oily liquids；
(3) with (2) gained, (1) gained oily liquids being concentrated oily liquids to merge, decompression rotation is evaporated, and obtains containing phosphorus
The protein coagulation thing 583kg containing phospholipid and unsaturated fatty acid of fat.
(4) with embodiment 1, surveying moisture in the protein coagulation thing containing phospholipid and unsaturated fatty acid that (3) obtain is
The 67% of gross weight, detects dried protein coagulation thing and obtains following index
| Composition | Account for the percentage ratio (%) of dry weight after drying |
| TL | 44.0 |
| Crude protein | 47.1 |
| Ash | 9.1 |
Further by the composition of fatty acid is analyzed, Agilent company of U.S. gas chromatogram 6890N is used to analyze, institute
The 122-7032 that analytical column is J&S Scientific DB-Wax used.Analyzing total fatty acid content in gained coagulum is
64.2%, wherein unsaturated fatty acid content is 34.8%(DHA7.6%, EPA content 14.6%).
(5) product is dissolved in 5 tons of dissolving tanks, configuration 3000L food pole organic solvent, wherein ethanol-normal hexane (4:1),
Using plate-and-frame filtration to remove insoluble substance, isolated liquid carries out Ultra filtration membrane again, deoils, solid-liquid ratio 1:20(w/v),
Film pressure 0.1Mpa, uses bore 100nm → 20nm bi-membrane method to separate.Prior to 55 DEG C, 100nm membrane filtration, backflow with ooze
Transparent liquid crosses 20nm film in 55 DEG C, and backflow and penetrating fluid volume ratio reach 1:3, collect 20nm film backflow, obtain unsaturated lipid
The concentrated solution of fat acid.
| Composition | Account for the percentage ratio (%) of dry weight after drying |
| TL | 51.8 |
| Crude protein | 39.9 |
| Ash | 8.3 |
Analyzing total fatty acid content in gained coagulum is 64.2%, and unsaturated fatty acid content is 34.8%(wherein DHA7.6%,
EPA content 14.6%).
(6) by concentrated solution flowing phase normal hexane: isopropanol: water (1:1:0.16, v/v/v) dissolves, stir under room temperature
Standby to dissolving.Learn from else's experience No. 2 silica gel of gross porosity of activation, add flowing and makes pulpous state mutually, wet method dress post, then equal with flowing
Weighing apparatus chromatographic column.The sample prepared slowly is poured in post, after sample introduction, adds flowing phase eluting.Controlling void tower flow velocity is
0.382cm/min, sample introduction concentration is under the room temperature condition of 0.25g concentrated phosphatide (mL/ flow phase), fixing selects 100 ~ 200 mutually
No. 2 silica gel of mesh gross porosity, purification rate is 70%, and the suitableeest upper carrying capacity is 0.086gPC(g/ silica gel), obtain PC content be 80% with
On phosphatidylcholine mixture.After lyophilization, obtain product phosphatidylcholines.
(7) in 3000L fermentation tank, the phosphatidylcholine 50kg of (6) gained, Serine 350kg, calcium chloride 6.0kg are put into
Reaction, adds water to 500L, then puts into PLD enzyme 1.6kg, after reacting 4h under the conditions of bath temperature 40-45 DEG C, pH5.6, then
Put into the PLD enzyme reaction 12h of 0.8kg.Mixture after reaction reduces pressure below 50 DEG C and removes solvent, (4) is obtained
Rich in the phosphatidylcholine race (PC) of many unsaturated double-bonds fatty acyl group, it is converted in aqueous phase reactions system rich in the most unsaturated
The Phosphatidylserine (PS) of double bond fatty acyl group, it is thus achieved that product 45.7kg, POPS therein (PS) content is about
39.3%。
(8) in the 45.7kg product (7) obtained, input 8000L, with in upper container, adds 1600L normal hexane and ethanol
400L, is subsequently adding the edible hydrochloric acid of 10-20L, is adjusted to pH3, is sufficiently stirred for, is layered, and removes upper liquid, at subnatant
In add 2000L normal hexane, stir, be layered, repeat to extract 3 times.Add water in the solution: ethanol (1:9, v/v)
150-300L, is thoroughly mixed, and regulates below pH9, separates hexane solution, and the product removing normal hexane carries out high-efficient liquid
Analysis of hplc, the content recording Phosphatidylserine (PS) is 45.1%.Utilize the PS product that the inventive method obtains, rich
Containing DHA and EPA, for natural prodcuts, activity is high, can add antioxidant, in order to protect as required in PS product
Deposit and prepare various health product and nutriment, medicine etc. with the later stage.
The extraction process of the phosphatidylcholine of antarctic krill of the present invention, specifically comprises the following steps that
A () collects Antarctic krill head, add water, and decocts, and krill head decoction liquor is delivered to solid-liquid separation in cold pressing expeller, obtained by stirring
Must be rich in the first-class solid of the oily liquids of phospholipid and krill；B () adds dehydrated alcohol in the first-class solid of krill, high shear extracts,
Centrifugation obtains supernatant；This step is carried out 1-2 time, merges supernatant, concentrates and obtains oily liquids；
C oily liquids in step (a) and step (b) is merged by (), drying under reduced pressure, it is thus achieved that containing phospholipid and unsaturated lipid
The protein coagulation thing of fat acid；
D protein coagulation thing containing phospholipid and unsaturated fatty acid is dissolved in food stage organic solvent by (), through flame filter press mistake
Filtering off except insoluble substance, the liquid obtained after filtration is again through membrane separation concentration, it is thus achieved that concentrated solution；
E () concentrated solution is through column chromatographic isolation and purification, it is thus achieved that phosphatidylcholine mixture；The most freeze-dried, it is thus achieved that phosphatidyl gallbladder
Alkali product.
The preparation method of the Phosphatidylserine of antarctic krill of the present invention, specifically comprises the following steps that
A () collects Antarctic krill head, boiling, stirring, krill head decoction liquor is delivered to solid-liquid separation in cold pressing expeller, it is thus achieved that
Oily liquids and the first-class solid of krill rich in phospholipid；
B () adds dehydrated alcohol in the first-class solid of krill, high shear extracts, and centrifugation obtains supernatant；This step extracts
1-2 time, merge supernatant, concentrate and obtain oily liquids；
C oily liquids in step (a) and step (b) is merged by (), drying under reduced pressure, it is thus achieved that containing phospholipid and unsaturated lipid
The protein coagulation thing of fat acid；
D protein coagulation thing containing phospholipid and unsaturated fatty acid is dissolved in food stage organic solvent by (), through flame filter press mistake
Filtering off except insoluble substance, the liquid obtained after filtration is again through membrane separation concentration, it is thus achieved that concentrated solution；
E () concentrated solution is through column chromatographic isolation and purification, it is thus achieved that phosphatidylcholine mixture；The most freeze-dried, it is thus achieved that phosphatidyl gallbladder
Alkali；
F (), in aqueous phase, adds Serine, Choline phosphatase, calcium chloride, hydrolytic phosphatide phatidylcholine, generates phosphatidyl silk ammonia
Acid；Remove solvent, it is thus achieved that the thick product of Phosphatidylserine；
G thick for Phosphatidylserine product is repeatedly extracted by () through 2-4 food stage organic solvent, adjusted by pH for the first time during extraction
Joint, to 3, removes organic solvent, it is thus achieved that Phosphatidylserine product.
In above-mentioned steps (f), below 50 DEG C, decompression method is used to remove solvent.
Can use micro-filtration membrane or ultrafilter membrane in above-mentioned membrance separation, the aperture of described film is 0.02-10 μm, applies pressure on film
0.015~10MPa。
Concentrated solution is first dissolved in by above-mentioned steps (e) in the flowing mutually of normal hexane, isopropanol, water composition, fixes and select mutually
50-400 mesh silochrom or macroporous resin.
During above-mentioned steps (e) column chromatography, described pillar blade diameter length ratio 1:5-10, described eluant selected from normal hexane aqueous solution,
The aqueous solution of isopropanol or normal hexane and the aqueous solution of isopropanol.For fish head, include but not limited to calamary head, it is also possible to reference
Technical scheme, extracts phosphatidylcholine, and decomposes phosphatidylcholine, it is thus achieved that rich in the Phosphatidylserine of DHA.
The present invention first passes through cold pressing and is extracted by the 60-70% in krill head phospholipid, then to the krill head solid obtained after cold pressing
Carry out high shear extraction, remaining 30-40% in krill head phospholipid is extracted, so the consumption of the dehydrated alcohol that extraction uses
Relatively conventional method is substantially reduced, and dehydrated alcohol can be with recycling use.
The present invention efficiently separates for Antarctic krill garbage and refines so that large-scale production high-load is raw from Antarctic krill
The industrialization producing Phosphatidylserine is possibly realized so that this product is possibly realized in the popularization of health food biomedicine field.
The above description of this invention is not limited to the scope of the present invention.For the person of ordinary skill of the art, may be used
Making various corresponding change with the technical scheme provided according to the present invention, these changes all should belong to protection scope of the present invention.
Claims (22)
1. the extraction process of the phosphatidylcholine of an antarctic krill, it is characterised in that comprise the steps:
A () collects Antarctic krill head, add water, and decocts, and krill head decoction liquor is delivered to solid-liquid separation in cold pressing expeller, it is thus achieved that rich in oily liquids and the first-class solid of krill of phospholipid by stirring；
B () adds dehydrated alcohol in the first-class solid of krill, high shear extracts, and centrifugation obtains supernatant；This step is carried out 1-2 time, merges supernatant, concentrates and obtains oily liquids；
C oily liquids in step (a) and step (b) is merged by (), drying under reduced pressure, it is thus achieved that the protein coagulation thing containing phospholipid and unsaturated fatty acid；
D protein coagulation thing containing phospholipid and unsaturated fatty acid is dissolved in food stage organic solvent by (), filter through flame filter press and remove insoluble substance, and the liquid obtained after filtration is again through membrane separation concentration, it is thus achieved that concentrated solution；Described food stage organic solvent is ethanol and normal hexane, described ethanol: the volume ratio of normal hexane is 2-4:0.5-1.5；
E () concentrated solution is through column chromatographic isolation and purification, it is thus achieved that phosphatidylcholine mixture；The most freeze-dried, it is thus achieved that product phosphatidylcholines.
2. the extraction process of the phosphatidylcholine of antarctic krill as claimed in claim 1, it is characterised in that in described step (a) in krill head 2h after thawing, add water, 40-55 DEG C of decoction, add ethanol during stirring.
3. the extraction process of the phosphatidylcholine of antarctic krill as claimed in claim 1, it is characterised in that in described step (b), supernatant is through concentrating under reduced pressure, it is thus achieved that oily liquids.
4. the extraction process of the phosphatidylcholine of antarctic krill as described in claim 1-3 any claim, it is characterised in that in described step (d), use Ultra filtration membrane, solid-liquid ratio W:V is 1:20, temperature 10-55 DEG C, uses bore 100nm → 20nm bi-membrane method to separate.
5. such as the extraction process of phosphatidylcholine of claim 1-3 any claim antarctic krill, it is characterized in that, in described step (d), using micro-filtration membrane or ultrafilter membrane in membrance separation, the aperture of described film is 0.02-10 μm, applies pressure 0.015 ~ 10MPa on film.
6. the extraction process of the phosphatidylcholine of antarctic krill as described in claim 1-3 any claim, it is characterized in that, concentrated solution is first dissolved in by described step (e) in the flowing mutually of normal hexane, isopropanol, water composition, fixing selection 50-400 mesh silochrom or macroporous resin mutually.
7. the extraction process of the phosphatidylcholine of antarctic krill as claimed in claim 6, it is characterized in that, during described step (e) column chromatography, described pillar blade diameter length ratio 1:5-10, described eluant is selected from the aqueous solution of normal hexane, the aqueous solution of isopropanol or normal hexane and the aqueous solution of isopropanol.
8. the extraction process of the phosphatidylcholine of antarctic krill as claimed in claim 6, it is characterised in that described fixing selection 100-200 mesh silochrom mutually.
9. the extraction process of the phosphatidylcholine of antarctic krill as claimed in claim 5, it is characterised in that in described step (d), using Ultra filtration membrane, solid-liquid ratio W:V is 1:20, temperature 10-55 DEG C, uses bore 100nm → 20nm bi-membrane method to separate.
10. the extraction process of the phosphatidylcholine of antarctic krill as described in claim 7 or 8, it is characterised in that normal hexane in described flowing mutually: isopropanol: water volume ratio is 1:1:0.16.
The preparation method of the Phosphatidylserine of 11. 1 kinds of antarctic krill, it is characterised in that comprise the steps:
A () collects Antarctic krill head, boiling, stirring, krill head decoction liquor is delivered to solid-liquid separation in cold pressing expeller, it is thus achieved that rich in oily liquids and the first-class solid of krill of phospholipid；
B () adds dehydrated alcohol in the first-class solid of krill, high shear extracts, and centrifugation obtains supernatant；This step extracts 1-2 time, merges supernatant, concentrates and obtains oily liquids；
C oily liquids in step (a) and step (b) is merged by (), drying under reduced pressure, it is thus achieved that the protein coagulation thing containing phospholipid and unsaturated fatty acid；
D protein coagulation thing containing phospholipid and unsaturated fatty acid is dissolved in food stage organic solvent by (), filter through flame filter press and remove insoluble substance, and the liquid obtained after filtration is again through membrane separation concentration, it is thus achieved that concentrated solution；
E () concentrated solution is through column chromatographic isolation and purification, it is thus achieved that phosphatidylcholine mixture；The most freeze-dried, it is thus achieved that phosphatidylcholine；
F (), in aqueous phase, adds Serine, Choline phosphatase, calcium chloride, hydrolytic phosphatide phatidylcholine, generates Phosphatidylserine；Remove solvent, it is thus achieved that the thick product of Phosphatidylserine；
G thick for Phosphatidylserine product is repeatedly extracted by () through 2-4 food stage organic solvent, for the first time by pH regulator to 3 during extraction, remove organic solvent, it is thus achieved that Phosphatidylserine product；
In described step (d) and (g), food stage organic solvent is ethanol and normal hexane, described ethanol: the volume ratio of normal hexane is 2-4:0.5-1.5.
The preparation method of the Phosphatidylserine of 12. antarctic krill as claimed in claim 11, it is characterised in that in described step (f), during hydrolysis, temperature is 40-45 DEG C, pH5-6, described phosphatidylcholine: Serine: Choline phosphatase is 1:7-13:0.03-0.06.
The preparation method of the Phosphatidylserine of 13. antarctic krill as claimed in claim 11, it is characterised in that in described step (a) in krill head 2h after thawing, cold pressing after boiling.
14. as described in claim 11-13 any claim the preparation method of the Phosphatidylserine of antarctic krill, it is characterized in that, in described step (g), for the first time during extraction, the mixture that food stage organic solvent is ethanol and normal hexane used, described ethanol and normal hexane volume ratio are 4:1, and use edible salt acid for adjusting pH to 3, remove upper liquid.
The preparation method of the Phosphatidylserine of 15. antarctic krill as claimed in claim 14, it is characterised in that in described step (g), uses normal hexane to carry out follow-up 1-3 extraction, and extraction terminates to remove upper liquid every time.
The preparation method of the Phosphatidylserine of 16. antarctic krill as claimed in claim 15, it is characterised in that in described step (g), repeatedly after extraction, uses 90% ethanol water, removes normal hexane, obtain Phosphatidylserine product.
17. such as the preparation method of the Phosphatidylserine of claim 11-13,15 or 16 antarctic krill as described in any claim, it is characterized in that, in described step (d), employing Ultra filtration membrane concentrates, solid-liquid ratio W:V is 1:20, temperature 10-55 DEG C, uses bore 100nm → 20nm bi-membrane method to separate.
18. such as the preparation method of the Phosphatidylserine of claim 11-13,15 or 16 antarctic krill as described in any claim, it is characterized in that, in described step (d), membrance separation uses micro-filtration membrane or ultrafilter membrane, the aperture of described film is 0.02-10 μm, applies pressure 0.015 ~ 10MPa on film.
19. such as the preparation method of the Phosphatidylserine of claim 11-13,15 or 16 antarctic krill as described in any claim, it is characterized in that, concentrated solution is first dissolved in by described step (e) in the flowing mutually of normal hexane, isopropanol, water composition, fixing selection 50-400 mesh silochrom or macroporous resin mutually.
The preparation method of the Phosphatidylserine of 20. antarctic krill as claimed in claim 18, it is characterized in that, concentrated solution is first dissolved in by described step (e) in the flowing mutually of normal hexane, isopropanol, water composition, fixing selection 50-400 mesh silochrom or macroporous resin mutually.
The preparation method of the Phosphatidylserine of 21. antarctic krill as claimed in claim 19, it is characterized in that, during described step (e) column chromatography, described pillar blade diameter length ratio 1:5-10, described eluant is selected from the aqueous solution of normal hexane, the aqueous solution of isopropanol or normal hexane and the aqueous solution of isopropanol.
22. claim 11-13,15,16, the preparation method of the Phosphatidylserine of antarctic krill as described in 20-21 any claim, it is characterised in that in described step (b), supernatant is through concentrating under reduced pressure, it is thus achieved that oily liquids.
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| CN105177073B (en) * | 2015-09-08 | 2016-06-08 | 威海博宇食品有限公司 | A kind of method preparing Phosphatidylserine |
| CN106083919B (en) * | 2016-06-11 | 2018-04-13 | 浙江工商大学 | Utilize the method for glycol-based silica gel extraction snakehead phosphatide |
| CN113662948B (en) * | 2021-09-10 | 2022-10-11 | 中国海洋大学 | Application of phospholipid and derivatives thereof in preparation of products for improving systemic lupus erythematosus |
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