CN102388988A - Separated extraction method of microorganism oil - Google Patents
Separated extraction method of microorganism oil Download PDFInfo
- Publication number
- CN102388988A CN102388988A CN2011103502006A CN201110350200A CN102388988A CN 102388988 A CN102388988 A CN 102388988A CN 2011103502006 A CN2011103502006 A CN 2011103502006A CN 201110350200 A CN201110350200 A CN 201110350200A CN 102388988 A CN102388988 A CN 102388988A
- Authority
- CN
- China
- Prior art keywords
- oil
- solid
- microbial oil
- surfactant solution
- branch
- 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.)
- Granted
Links
- 244000005700 microbiome Species 0.000 title abstract description 11
- 238000000605 extraction Methods 0.000 title abstract 3
- 239000004094 surface-active agent Substances 0.000 claims abstract description 89
- 239000007787 solid Substances 0.000 claims abstract description 75
- 238000000034 method Methods 0.000 claims abstract description 49
- 239000013078 crystal Substances 0.000 claims abstract description 47
- 239000004519 grease Substances 0.000 claims abstract description 44
- 239000007788 liquid Substances 0.000 claims abstract description 42
- 239000002002 slurry Substances 0.000 claims abstract description 34
- 238000003756 stirring Methods 0.000 claims abstract description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 23
- 239000000839 emulsion Substances 0.000 claims abstract description 18
- 238000002425 crystallisation Methods 0.000 claims abstract description 10
- 230000008025 crystallization Effects 0.000 claims abstract description 7
- 238000009413 insulation Methods 0.000 claims abstract description 7
- 230000000813 microbial effect Effects 0.000 claims description 89
- 239000012530 fluid Substances 0.000 claims description 38
- 150000003839 salts Chemical class 0.000 claims description 24
- 230000008014 freezing Effects 0.000 claims description 16
- 238000007710 freezing Methods 0.000 claims description 16
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 14
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 14
- 230000004927 fusion Effects 0.000 claims description 14
- 239000007790 solid phase Substances 0.000 claims description 14
- 229910052938 sodium sulfate Inorganic materials 0.000 claims description 11
- 235000011152 sodium sulphate Nutrition 0.000 claims description 11
- 239000000126 substance Substances 0.000 claims description 10
- 239000000470 constituent Substances 0.000 claims description 9
- 230000001804 emulsifying effect Effects 0.000 claims description 8
- 239000012071 phase Substances 0.000 claims description 8
- -1 sodium alkyl benzene Chemical class 0.000 claims description 8
- 238000010792 warming Methods 0.000 claims description 8
- RCEAADKTGXTDOA-UHFFFAOYSA-N OS(O)(=O)=O.CCCCCCCCCCCC[Na] Chemical group OS(O)(=O)=O.CCCCCCCCCCCC[Na] RCEAADKTGXTDOA-UHFFFAOYSA-N 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 7
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 7
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 7
- 239000011734 sodium Substances 0.000 claims description 7
- 229910052708 sodium Inorganic materials 0.000 claims description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 229940077388 benzenesulfonate Drugs 0.000 claims description 4
- DIZPMCHEQGEION-UHFFFAOYSA-H aluminium sulfate (anhydrous) Chemical compound [Al+3].[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O DIZPMCHEQGEION-UHFFFAOYSA-H 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- DCXXMTOCNZCJGO-UHFFFAOYSA-N tristearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCCCC DCXXMTOCNZCJGO-UHFFFAOYSA-N 0.000 claims 2
- 239000006228 supernatant Substances 0.000 claims 1
- 238000001816 cooling Methods 0.000 abstract description 14
- 238000004140 cleaning Methods 0.000 abstract description 3
- 238000001035 drying Methods 0.000 abstract 1
- 238000000926 separation method Methods 0.000 abstract 1
- 239000003921 oil Substances 0.000 description 126
- 239000000243 solution Substances 0.000 description 48
- 239000003925 fat Substances 0.000 description 30
- 235000019197 fats Nutrition 0.000 description 30
- 241000894006 Bacteria Species 0.000 description 11
- YZXBAPSDXZZRGB-DOFZRALJSA-N arachidonic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O YZXBAPSDXZZRGB-DOFZRALJSA-N 0.000 description 10
- 230000008569 process Effects 0.000 description 10
- 150000002632 lipids Chemical class 0.000 description 9
- 239000002904 solvent Substances 0.000 description 9
- 238000004519 manufacturing process Methods 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 230000015556 catabolic process Effects 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 235000021342 arachidonic acid Nutrition 0.000 description 5
- 229940114079 arachidonic acid Drugs 0.000 description 5
- 238000009472 formulation Methods 0.000 description 5
- 235000020978 long-chain polyunsaturated fatty acids Nutrition 0.000 description 5
- 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 4
- 238000005516 engineering process Methods 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 235000020673 eicosapentaenoic acid Nutrition 0.000 description 3
- 229960005135 eicosapentaenoic acid Drugs 0.000 description 3
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 235000012000 cholesterol Nutrition 0.000 description 2
- 230000015271 coagulation Effects 0.000 description 2
- 238000005345 coagulation Methods 0.000 description 2
- 235000014113 dietary fatty acids Nutrition 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 229930195729 fatty acid Natural products 0.000 description 2
- 150000004665 fatty acids Chemical class 0.000 description 2
- 230000035800 maturation Effects 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- 230000000050 nutritive effect Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 238000009736 wetting Methods 0.000 description 2
- 235000019737 Animal fat Nutrition 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 239000004215 Carbon black (E152) Substances 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- SBJKKFFYIZUCET-JLAZNSOCSA-N Dehydro-L-ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(=O)C1=O SBJKKFFYIZUCET-JLAZNSOCSA-N 0.000 description 1
- PVNIQBQSYATKKL-UHFFFAOYSA-N Glycerol trihexadecanoate Natural products CCCCCCCCCCCCCCCC(=O)OCC(OC(=O)CCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCC PVNIQBQSYATKKL-UHFFFAOYSA-N 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 229940121363 anti-inflammatory agent Drugs 0.000 description 1
- 239000002260 anti-inflammatory agent Substances 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000002490 cerebral effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- MBMBGCFOFBJSGT-KUBAVDMBSA-N docosahexaenoic acid Natural products CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCC(O)=O MBMBGCFOFBJSGT-KUBAVDMBSA-N 0.000 description 1
- 235000020669 docosahexaenoic acid Nutrition 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 235000021149 fatty food Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 210000001640 nerve ending Anatomy 0.000 description 1
- 210000004126 nerve fiber Anatomy 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 235000014593 oils and fats Nutrition 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 230000004491 retinal development Effects 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 238000005204 segregation Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 235000019871 vegetable fat Nutrition 0.000 description 1
Images
Landscapes
- Fats And Perfumes (AREA)
Abstract
The invention relates to a separated extraction method of microorganism oil, which is characterized by comprising the following steps: firstly, homogenizing, stirring microorganism oil to be in separated extraction uniformly; secondly, breaking crystals; thirdly, cooling and crystallizing, crystallizing the heat-insulation microorganism oil for 4-7h at the temperature being 4 DEG C, and then arranging the heat-insulation microorganism oil for 1-4h at the low temperature being 10 DEG C, thus obtaining thick slurry containing solid crystallization; fourthly, adding surface active agent water solution in the thick slurry containing solid crystallization, and stirring, thus obtaining elusion system; fifthly, conducting centrifugal separation to obtain liquid oil, standing the emulsion system for 1-5h at the low temperature being 5-10 DEG C, centrifuging the emulsion system in a cooling centrifuge, separating liquid components in a light phase mode, putting the liquid components in light phase in water for cleaning, and dehydrating and drying the liquid components in light phase, thus obtaining the liquid oil of the microorganism oil; and sixthly, breaking the emulsion to obtain solid grease. The liquid oil of the microorganism separated and extracted by the method is not easy to solidify at lower temperature, the method procedure is simple and the cost is low.
Description
Technical field
The invention belongs to the Food Science field, be specifically related to a kind of branch extracting method of microbial oil.
Background technology
Microbial grease is another the human consumption grease new resources that after vegetable fat, animal fat, develops.Microbial grease is claimed Unicell Oils and Fats again, is that to utilize carbohydrate, hydrocarbon and common grease under certain condition be carbon source, nitrogenous source, be aided with grease and other lipids with commercial value of production of inorganic by microorganisms such as yeast, mould, bacterium and algae.The micro-organisms grease not only have the fat content height, with short production cycle, do not receive seasonal effect, advantage such as do not occupy cultivated land; And method such as available Fusion of Cells, cell mutation; Make the grease of microorganisms high-nutrition oil fat or some special fatty acid composition, as contain the functional grease of arachidonic acid (ARA), eicosapentaenoic acid (EPA), DHA long-chain polyunsaturated fatty acids (LC-PUFAs) such as (DHA).LC-PUFAs has tangible effect at aspects such as angiocardiopathy preventing, reducing blood lipid, reduction cholesterol, fat-reducing, inhibition tumor growth, anti-inflammatories.Since the nineties in 20th century, the production that the development and use microorganism carries out functional grease has become a big focus, as utilizes microbe culture to produce the research that ARA, EPA, DHA etc. are of high nutritive value and have the function grease of special health care.ARA belongs to the serial long-chain polyunsaturated fatty acid of ω-6 (LC-PUFA); It distributes in human body extensively; Content generally accounts for 40%～50% of polyunsaturated fatty acid total amount in brain and nerve fiber; At nerve endings more up to 70%, to cerebral function and the retinal development material that is absolutely necessary; In addition, ARA also has the effect of cholesterol in the function that promotes to grow and hypoglycemic, reducing blood lipid and the reduction blood.
But microbial oil can occur solidifying when low tempertaure storage, just can become solid-state or semisolid like temperature in the winter time when low, not only causes the oil product poor mobile performance, and inconvenience is used, and is difficult for storage; The transparency of oil product is reduced, the palatability variation, thus reduced the quality of the nutritive value and the fatty foods of microbial oil.It is according to different fatty acid triglycercide fusing point differences in the grease that the grease branch is carried, and makes high melting point component produce crystallization through cooling, obtains the different component of fusing point through filtering or centrifugalizing.The grease branch is carried and can be classified into single relatively product with forming complicated natural oil; Realize the fluid oil of different melting points and separating of solid fat; To improve the conformability of grease to different instructions for uses, enlarge the purposes of grease, improve the use value and the economic worth of grease.
At present the grease branch is put forward technology and is mainly contained that the dry method branch is carried, the solvent branch is carried and the surfactant branch is carried.The dry method branch is carried and is meant and does not add any solvent, and the grease that will be in dissolved state slowly is cooled to a certain degree, and the method for solid ester is separated out in isolated by filtration crystallization then.The dry method branch is carried and is divided into three steps: heat treated; Form crystal atoms nuclear and let crystal growing and maturation with cooling method; From solid, separate palmitin with filtering.Though the dry method branch has been put forward its unique superior property, it is low that the dry method branch is put forward efficient, and liquid oil contents is higher in the solid-state fat, makes the brilliant level of solid-state fat and fluid oil low.Solvent branch formulation is meant that in grease, adding a certain solvent in proportion forms the miscella system, carries out crystallisation by cooling then, divides a kind of minute extracting method of carrying.Solvent branch formulation can form the stable crystalline of easy filtration, improves separating effect, increases isolated yield, shortens disengaging time, improves the purity of products of separated.Solvent branch formulation branch is carried the efficient height, and solid-state fat constituent mass is good.Yet because crystallization temperature is low and divide to carry and relate to solvent loss in the process, its investment is bigger, and producing cost is high, is used as the hexane of solvent, acetone, and isopropyl alcohols etc. have inflammability.Surfactant branch formulation is meant behind the grease crystallisation by cooling, adds surfactant, improves the interfacial tension of oil and fat, borrows the affinity between fat and surfactant, forms the suspension of fat in aqueous surfactant solution, promotes the method for the brilliant segregation of fat.Operations such as its technology comprises that crystallisation by cooling, surfactant are moistening, centrifugation and surfactant recovery.The surfactant method branch is carried the separative efficiency height, superior product quality, and purposes is wide, is applicable to large-scale production.
Summary of the invention
Be prone to coagulation problem in order to solve microbial oil (10-20 ℃) when temperature is low; The branch extracting method that the purpose of this invention is to provide a kind of microbial oil; This method divides the fluid oil (10-20 ℃) when temperature is low of the microbial oil that proposes to be difficult for solidifying, and this method operation is simple, cost is low.
To achieve these goals, technical scheme of the present invention is: a kind of branch extracting method of microbial oil is characterized in that it comprises the steps:
1) homogenizing: will treat that the microbial oil (or claiming microbial grease) that branch is carried stirs;
2) break crystalline substance: after the microbial oil that stirs is warming up to 25-30 ℃, insulation 5-10min;
Microbial grease destroys the inhomogeneous nucleus that forms naturally before freezing, and microbial grease is separated out crystal owing to the microbial grease temperature is lower than solid-state fat freezing point in refining, transportation, storage process; This part crystal is owing to separate out in non-at the uniform velocity temperature-fall period, and crystal formation is different, and grain size differs; When forwarding freezing and crystallizing to after the stage; Can be unfavorable for even growth and maturation that fat is brilliant, make crystalline solid itself produce defective, influence the grease branch and carry; Broken crystalline substance is warming up to the grease fusion more than the solid-state fat fusing point usually, keeps 5-10min;
3) freezing and crystallizing (process of cooling gradually): with step 2) microbial oil after the insulation is in 4 ℃ of crystallization 4-7h (cryogenic conditions down fully); Place 1-4h under 10 ℃ of cryogenic conditions again, obtain containing the viscous slurry (having formed the viscous slurry that contains solid crystal after lipid material process cooling this moment is freezing) of solid crystal;
4) adding of surfactant solution: by the viscous slurry that contains solid crystal: the volume ratio of aqueous surfactant solution=1: 0.5-1: 2, choose the viscous slurry and the aqueous surfactant solution that contain solid crystal; Wherein, aqueous surfactant solution is made up of surfactant, inorganic salts and water, and surfactant concentrations is 0.08-0.4wt%, and the concentration of inorganic salts is 0.5-3wt%;
In the viscous slurry that contains solid crystal, add aqueous surfactant solution, stir 0.5-1.0h, obtain the emulsion system;
Fully stirring is uniformly dispersed mixed system and forms the emulsion system; Liquid component in the grease maintenance liquid phase that carries on as before; Can not be wetting and under stirring action, become fine droplets and be scattered in the aqueous solution by surfactant; And fully wetting by the crystalline particle of surfactant molecule parcel because of obtaining, got rid of the liquid component that invests crystal plane, so be transferred to aqueous phase;
5) centrifugalize liquid oil: the emulsifying liquid of step 4) is lain in 5-10 ℃ of cryogenic conditions leaves standstill 0.5-5h; Centrifugal in refrigerated centrifuge; Liquid component adds the entry cleaning after dehydrate (20-25 ℃ of dry 2-6h) just gently to tell mutually, promptly gets the fluid oil (product) of microbial oil;
The aqueous surfactant solution of solid crystal of suspending is then told with heavy phase, obtains solid phase grease mixed liquor.
Surfactant is lauryl sodium sulfate (SDS), sodium alkyl benzene sulfonate or sodium alkyl sulfate etc.
Inorganic salts are magnesium sulfate (MgSO
4), sodium sulphate, aluminum sulfate or sodium chloride etc.
The condition of the described centrifugation of step 5) is: 3000-6000rpm, centrifugal 5-10min.
6) get solid fat behind the breakdown of emulsion: after taking out the fluid oil (being liquid component) of microbial oil; The solid phase grease mixed liquor of gained is handled 2-10min through 80-95 ℃ of heating demulsification type, watery fusion, and solid constituent is promptly separated with water; The water that removes sub-cloud promptly obtains the solid fat component; Add entry and wash the aqueous surfactant solution of removing remnants, after dehydrating, the solid that promptly gets microbial oil is stearic.
Branch of the present invention is carried the fluid oil (being the product that will obtain required for the present invention) of back microbial bacteria oil, and clearing time is 120h above (its physical and chemical index is seen table 1) under 10 ℃ of conditions.This invention has solved microbial oil (＜20 ℃) when temperature is low and has been prone to coagulation problem, thereby has improved the use value and the economic worth of microbial grease.
The comparison of table 1 physical and chemical index (embodiment 1-5)
The invention has the beneficial effects as follows:
1) this method divides the fluid oil (10-20 ℃) when temperature is low of the microbial oil that proposes to be difficult for solidifying.Branch of the present invention is carried the fluid oil (being the product that will obtain required for the present invention) of back microbial bacteria oil, and clearing time is more than the 120h under 10 ℃ of conditions.
The fluid oil fusing point of the microbial oil that after branch is carried, obtains is low, and frost resistance is strong, and still keep liquid winter in south, can be used as the ready-mixed oil component and also can change microbial oil awkward situation in winter, enriches grease market.
2) this method operation is simple, does not need complex device.
The present invention and dry method branch are carried comparing to have with short production cyclely, and separative efficiency is high, the advantage that production process is easy to control; Comparing with solvent branch formulation does not need with an organic solvent, thereby safety, and equipment investment is few, and cost is lower.
3) fluid oil of microbial oil (product) better quality, the yield of liquid oil has no the pollution of organic solvent than higher;
4) used surfactant can reclaim the back repeated use in the technology;
5) owing to do not need complex device and need not use organic solvent, save great deal of investment, operating cost, thereby reduced production cost, be applicable to suitability for industrialized production.
Description of drawings
Fig. 1 is a process chart of the present invention.
The specific embodiment
In order to understand the present invention better, further illustrate content of the present invention below in conjunction with embodiment, but content of the present invention not only is confined to following embodiment.
Embodiment 1:
As shown in Figure 1, a kind of branch extracting method of microbial oil, it comprises the steps:
1) homogenizing: will treat that the microbial oil that branch is carried stirs;
2) broken brilliant: as the microbial oil fusion that stirs to be warming up to (25 ℃) more than the solid-state fat fusing point, to keep 5min;
3) the microbial oil sufficient crystallising 5h under 4 ℃ of cryogenic conditions after insulation freezing and crystallizing: with step 2); Place 10 ℃ of cryogenic thermostat water-bath 2h again, obtain containing the viscous slurry (having formed the viscous slurry that contains solid crystal after lipid material process cooling this moment is freezing) of solid crystal;
4) adding of surfactant solution: by the viscous slurry that contains solid crystal: the volume ratio of aqueous surfactant solution=1: 1, choose the viscous slurry and the aqueous surfactant solution that contain solid crystal; Wherein, aqueous surfactant solution is made up of surfactant, inorganic salts and water, and surfactant concentrations is 0.4wt%, and the concentration of inorganic salts is 1wt%;
Surfactant is lauryl sodium sulfate (SDS); Inorganic salts are magnesium sulfate (MgSO
4);
In the viscous slurry that contains solid crystal, add aqueous surfactant solution, stir 0.5h, obtain the emulsion system;
5) centrifugalize liquid oil: the emulsifying liquid of step 4) is lain in 10 ℃ of cryogenic conditions leaves standstill 0.5h; Centrifugal in refrigerated centrifuge; Liquid component is just gently to tell (abbreviating liquid oil among Fig. 1 as) mutually; Add the entry cleaning after dehydrate (20 ℃ of dry 6h), promptly get the fluid oil (product, yield are 59.9%) of microbial oil;
The aqueous surfactant solution of solid crystal of suspending is then told (being solid phase grease mixed liquor) with heavy phase;
6) get solid fat behind the breakdown of emulsion: after taking out the fluid oil (being liquid component) of microbial oil; The solid phase grease mixed liquor of gained is handled 2-10min through 90 ℃ of heating demulsification types, watery fusion, and solid constituent is promptly separated with water; The water that removes sub-cloud promptly obtains the solid fat component; Add entry and wash the aqueous surfactant solution of removing remnants, after dehydrating, promptly get the solid stearic (abbreviating solid fat among Fig. 1 as) of microbial oil.Surplus solution is a surfactant solution, after reclaiming, utilizes again.
The present embodiment branch is carried the fluid oil of back microbial bacteria oil, and clearing time is 120h above (its physical and chemical index is seen table 1) under 10 ℃ of conditions.The fluid oil (10-20 ℃) when temperature is low that the microbial oil that this method branch proposes is described is difficult for solidifying.
Embodiment 2:
A kind of branch extracting method of microbial oil, it comprises the steps:
1) homogenizing: will treat that the microbial oil that branch is carried stirs;
2) broken brilliant: as the microbial oil fusion that stirs to be warming up to (30 ℃) more than the solid-state fat fusing point, to keep 5min;
3) freezing and crystallizing: with microbial oil sufficient crystallising 5h under 4 ℃ of cryogenic conditions; Place 10 ℃ of cryogenic thermostat water-bath 2h again, obtain containing the viscous slurry (having formed the viscous slurry that contains solid crystal after lipid material process cooling this moment is freezing) of solid crystal;
4) adding of surfactant solution: by the viscous slurry that contains solid crystal: the volume ratio of aqueous surfactant solution=1: 1, choose the viscous slurry and the aqueous surfactant solution that contain solid crystal; Wherein, aqueous surfactant solution is made up of surfactant, inorganic salts and water, and surfactant concentrations is 0.4wt%, and the concentration of inorganic salts is 2wt%;
Surfactant is lauryl sodium sulfate (SDS); Inorganic salts are magnesium sulfate (MgSO
4);
In the viscous slurry that contains solid crystal, add aqueous surfactant solution, stir 1.0h, obtain the emulsion system;
5) centrifugalize liquid oil: the emulsifying liquid of step 4) is lain in 10 ℃ of cryogenic conditions leaves standstill 1h; Centrifugal in refrigerated centrifuge, liquid component adds entry and cleans after dehydrate (25 ℃ of dry 6h) just gently to tell mutually; Promptly get the fluid oil (product, yield are 57.1%) of microbial oil;
The aqueous surfactant solution of solid crystal of suspending is then told (being solid phase grease mixed liquor) with heavy phase;
6) get solid fat behind the breakdown of emulsion: after taking out the fluid oil (being liquid component) of microbial oil; The solid phase grease mixed liquor of gained is handled 2-10min through 90 ℃ of heating demulsification types, watery fusion, and solid constituent is promptly separated with water; The water that removes sub-cloud promptly obtains the solid fat component; Add entry and wash the aqueous surfactant solution of removing remnants, after dehydrating, the solid that promptly gets microbial oil is stearic.
The present embodiment branch is carried the fluid oil of back microbial bacteria oil, and clearing time is 120h above (its physical and chemical index is seen table 1) under 10 ℃ of conditions.The fluid oil (10-20 ℃) when temperature is low that the microbial oil that this method branch proposes is described is difficult for solidifying.
Embodiment 3:
A kind of branch extracting method of microbial oil, it comprises the steps:
1) homogenizing: will treat that the microbial oil that branch is carried stirs;
2) broken brilliant: as the microbial oil fusion that stirs to be warming up to (28 ℃) more than the solid-state fat fusing point, to keep 5min;
3) freezing and crystallizing: with microbial oil sufficient crystallising 5h under 4 ℃ of cryogenic conditions, place 10 ℃ of cryogenic thermostat water-bath 2h again, formed the viscous slurry that contains solid crystal after lipid material process cooling this moment is freezing;
4) adding of surfactant solution: by the viscous slurry that contains solid crystal: the volume ratio of aqueous surfactant solution=1: 1.5, choose the viscous slurry and the aqueous surfactant solution that contain solid crystal; Wherein, aqueous surfactant solution is made up of surfactant, inorganic salts and water, and surfactant concentrations is 0.4wt%, and the concentration of inorganic salts is 1wt%;
Surfactant is lauryl sodium sulfate (SDS); Inorganic salts are magnesium sulfate (MgSO
4);
In the viscous slurry that contains solid crystal, add aqueous surfactant solution, stir 0.8h, obtain the emulsion system;
5) centrifugalize liquid oil: the emulsifying liquid of step 4) is lain in 10 ℃ of cryogenic conditions leaves standstill 2h; Centrifugal in refrigerated centrifuge, liquid component adds entry and cleans after dehydrate (22 ℃ of dry 4h) just gently to tell mutually; Promptly get the fluid oil (product, 56.8%) of microbial oil;
The aqueous surfactant solution of solid crystal of suspending is then told (being solid phase grease mixed liquor) with heavy phase;
6) get solid fat behind the breakdown of emulsion: after taking out the fluid oil (being liquid component) of microbial oil; The solid phase grease mixed liquor of gained is handled 2-10min through 80 ℃ of heating demulsification types, watery fusion, and solid constituent is promptly separated with water; The water that removes sub-cloud promptly obtains the solid fat component; Add entry and wash the aqueous surfactant solution of removing remnants, after dehydrating, the solid that promptly gets microbial oil is stearic.
The present embodiment branch is carried the fluid oil of back microbial bacteria oil, and clearing time is 120h above (its physical and chemical index is seen table 1) under 10 ℃ of conditions.The fluid oil (10-20 ℃) when temperature is low that the microbial oil that this method branch proposes is described is difficult for solidifying.
Embodiment 4:
A kind of branch extracting method of microbial oil, it comprises the steps:
1) homogenizing: will treat that the microbial oil that branch is carried stirs;
2) broken brilliant: as the microbial oil fusion that stirs to be warming up to (26 ℃) more than the solid-state fat fusing point, to keep 5min;
3) freezing and crystallizing: with microbial oil sufficient crystallising 4h under 4 ℃ of cryogenic conditions, place 10 ℃ of cryogenic thermostat water-bath 1h again, formed the viscous slurry that contains solid crystal after lipid material process cooling this moment is freezing;
4) adding of surfactant solution: by the viscous slurry that contains solid crystal: the volume ratio of aqueous surfactant solution=1: 0.5, choose the viscous slurry and the aqueous surfactant solution that contain solid crystal; Wherein, aqueous surfactant solution is made up of surfactant, inorganic salts and water, and surfactant concentrations is 0.08wt%, and the concentration of inorganic salts is 0.5wt%;
Surfactant is lauryl sodium sulfate (SDS); Inorganic salts are magnesium sulfate (MgSO
4);
In the viscous slurry that contains solid crystal, add aqueous surfactant solution, stir 0.6h, obtain the emulsion system;
5) centrifugalize liquid oil: the emulsifying liquid of step 4) is lain in 5 ℃ of cryogenic conditions leaves standstill 1h; Centrifugal in refrigerated centrifuge, liquid component adds entry and cleans after dehydrate (20 ℃ of dry 2h) just gently to tell mutually; Promptly get the fluid oil (product, yield are 58.8%) of microbial oil;
The aqueous surfactant solution of solid crystal of suspending is then told (being solid phase grease mixed liquor) with heavy phase;
6) get solid fat behind the breakdown of emulsion: after taking out the fluid oil (being liquid component) of microbial oil; The solid phase grease mixed liquor of gained is handled 2-10min through 85 ℃ of heating demulsification types, watery fusion, and solid constituent is promptly separated with water; The water that removes sub-cloud promptly obtains the solid fat component; Add entry and wash the aqueous surfactant solution of removing remnants, after dehydrating, the solid that promptly gets microbial oil is stearic.
The present embodiment branch is carried the fluid oil of back microbial bacteria oil, and clearing time is 120h above (its physical and chemical index is seen table 1) under 10 ℃ of conditions.The fluid oil (10-20 ℃) when temperature is low that the microbial oil that this method branch proposes is described is difficult for solidifying.
Embodiment 5:
A kind of branch extracting method of microbial oil, it comprises the steps:
1) homogenizing: will treat that the microbial oil that branch is carried stirs;
2) broken brilliant: as the microbial oil fusion that stirs to be warming up to (30 ℃) more than the solid-state fat fusing point, to keep 10min;
3) freezing and crystallizing: with microbial oil sufficient crystallising 7h under 4 ℃ of cryogenic conditions; Place 10 ℃ of cryogenic thermostat water-bath 4h again, obtain containing the viscous slurry (having formed the viscous slurry that contains solid crystal after lipid material process cooling this moment is freezing) of solid crystal;
4) adding of surfactant solution: in this slip, contain 0.3%SDS and 1%MgSO according to the adding of 1: 1 ratio of oil-water ratio
4Aqueous surfactant solution, fully stir and mixed system be uniformly dispersed form the emulsion system;
5) centrifugalize liquid oil: this emulsifying liquid is lain in 10 ℃ of conditions leaves standstill 0.5h; In refrigerated centrifuge with 5000rpm; Centrifugal 5min; Liquid component just gently to tell mutually, adds washed with de-ionized water after dehydrate the fluid oil (yield is 57.2%) that promptly gets microbial oil, and the aqueous surfactant solution of the solid crystal that suspending is then told with heavy phase;
4) adding of surfactant solution: by the viscous slurry that contains solid crystal: the volume ratio of aqueous surfactant solution=1: 2, choose the viscous slurry and the aqueous surfactant solution that contain solid crystal; Wherein, aqueous surfactant solution is made up of surfactant, inorganic salts and water, and surfactant concentrations is 0.2wt%, and the concentration of inorganic salts is 3wt%;
Surfactant is lauryl sodium sulfate (SDS); Inorganic salts are magnesium sulfate (MgSO
4);
In the viscous slurry that contains solid crystal, add aqueous surfactant solution, stir 1.0h, obtain the emulsion system;
5) centrifugalize liquid oil: the emulsifying liquid of step 4) is lain in 8 ℃ of cryogenic conditions leaves standstill 5h; Centrifugal in refrigerated centrifuge, liquid component adds entry and cleans after dehydrate (25 ℃ of dry 2h) just gently to tell mutually; Promptly get the fluid oil (product, yield are 57.2%) of microbial oil;
The aqueous surfactant solution of solid crystal of suspending is then told (being solid phase grease mixed liquor) with heavy phase;
6) get solid fat behind the breakdown of emulsion: after taking out the fluid oil (being liquid component) of microbial oil; The solid phase grease mixed liquor of gained is handled 2-10min through 95 ℃ of heating demulsification types, watery fusion, and solid constituent is promptly separated with water; The water that removes sub-cloud promptly obtains the solid fat component; Add entry and wash the aqueous surfactant solution of removing remnants, after dehydrating, the solid that promptly gets microbial oil is stearic.
The present embodiment branch is carried the fluid oil of back microbial bacteria oil, and clearing time is 120h above (its physical and chemical index is seen table 1) under 10 ℃ of conditions.The fluid oil (10-20 ℃) when temperature is low that the microbial oil that this method branch proposes is described is difficult for solidifying.
Embodiment 6
Basic identical with embodiment 1, difference is: surfactant is a sodium alkyl benzene sulfonate.Inorganic salts are sodium sulphate.
The present embodiment branch is carried the fluid oil of back microbial bacteria oil, and clearing time is more than the 120h under 10 ℃ of conditions.The fluid oil (10-20 ℃) when temperature is low that the microbial oil that this method branch proposes is described is difficult for solidifying.
Embodiment 7
Basic identical with embodiment 1, difference is: surfactant is a sodium alkyl sulfate.Inorganic salts are aluminum sulfate.
The present embodiment branch is carried the fluid oil of back microbial bacteria oil, and clearing time is more than the 120h under 10 ℃ of conditions.The fluid oil (10-20 ℃) when temperature is low that the microbial oil that this method branch proposes is described is difficult for solidifying.
Embodiment 7
Basic identical with embodiment 1, difference is: surfactant is a sodium alkyl benzene sulfonate.Inorganic salts are sodium chloride.
The present embodiment branch is carried the fluid oil of back microbial bacteria oil, and clearing time is more than the 120h under 10 ℃ of conditions.The fluid oil (10-20 ℃) when temperature is low that the microbial oil that this method branch proposes is described is difficult for solidifying.
The bound value of each technological parameter of the present invention (like temperature, time etc.), with and interval value, can both realize the present invention, do not enumerate embodiment one by one at this.
Claims (4)
1. the branch extracting method of a microbial oil is characterized in that it comprises the steps:
1) homogenizing: will treat that the microbial oil that branch is carried stirs;
2) break crystalline substance: after the microbial oil that stirs is warming up to 25-30 ℃, insulation 5-10min;
3) freezing and crystallizing: with step 2) microbial oil after the insulation places 1-4h under 10 ℃ of cryogenic conditions again in 4 ℃ of crystallization 4-7h, obtains containing the viscous slurry of solid crystal;
4) adding of surfactant solution: by the viscous slurry that contains solid crystal: the volume ratio of aqueous surfactant solution=1: 0.5-1: 2, choose the viscous slurry and the aqueous surfactant solution that contain solid crystal; Wherein, aqueous surfactant solution is made up of surfactant, inorganic salts and water, and surfactant concentrations is 0.08-0.4wt%, and the concentration of inorganic salts is 0.5-3wt%;
In the viscous slurry that contains solid crystal, add aqueous surfactant solution, stir 0.5-1.0h, obtain the emulsion system;
5) centrifugalize liquid oil: the emulsifying liquid of step 4) is lain in 5-10 ℃ of cryogenic conditions leave standstill 1-5h, centrifugal in refrigerated centrifuge, liquid component is just gently to tell mutually, adds entry and cleans after dehydrate, and promptly gets the fluid oil of microbial oil;
The aqueous surfactant solution of solid crystal of suspending is then told with heavy phase, obtains solid phase grease mixed liquor.
2. the branch extracting method of a kind of microbial oil according to claim 1, it is characterized in that: surfactant is lauryl sodium sulfate (SDS), sodium alkyl benzene sulfonate or sodium alkyl sulfate.
3. the branch extracting method of a kind of microbial oil according to claim 1, it is characterized in that: inorganic salts are magnesium sulfate (MgSO
4), sodium sulphate, aluminum sulfate or sodium chloride.
4. the branch extracting method of a kind of microbial oil according to claim 1; It is characterized in that: the solid phase grease mixed liquor of gained is handled 2-10min through 80-95 ℃ of heating demulsification type, watery fusion, and solid constituent is promptly separated with water; Get supernatant; The solid constituent that obtains also adds entry and washes and remove remaining aqueous surfactant solution, after dehydrating, promptly get the solid tristearin of microbial oil.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110350200 CN102388988B (en) | 2011-11-08 | 2011-11-08 | Separated extraction method of microorganism oil |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110350200 CN102388988B (en) | 2011-11-08 | 2011-11-08 | Separated extraction method of microorganism oil |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102388988A true CN102388988A (en) | 2012-03-28 |
| CN102388988B CN102388988B (en) | 2013-01-23 |
Family
ID=45856483
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110350200 Expired - Fee Related CN102388988B (en) | 2011-11-08 | 2011-11-08 | Separated extraction method of microorganism oil |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102388988B (en) |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103421605A (en) * | 2012-05-25 | 2013-12-04 | 丰益(上海)生物技术研发中心有限公司 | Method for fractionating grease through using erythritol fatty acid ester, and application of erythritol fatty acid ester |
| CN105176671A (en) * | 2014-06-18 | 2015-12-23 | 中国石油化工股份有限公司 | Method for wall breaking of oil-containing microalgae and assistant assisted extraction of oil |
| CN105454466A (en) * | 2015-12-04 | 2016-04-06 | 润科生物工程（福建）有限公司 | Application of sucrose fatty acid ester serving as grease crystallization inhibitor in oil containing polyunsaturated fatty acid single-cell grease |
| CN108624403A (en) * | 2018-04-28 | 2018-10-09 | 武汉工程大学 | A kind of method of low temperature non-organic solvent extraction energy insect oils |
| US10329515B2 (en) | 2000-01-19 | 2019-06-25 | Dsm Ip Assets B.V. | Solventless extraction process |
| US10342772B2 (en) | 2013-12-20 | 2019-07-09 | Dsm Ip Assets B.V. | Processes for obtaining microbial oil from microbial cells |
| US10364207B2 (en) | 2013-12-20 | 2019-07-30 | Dsm Ip Assets B.V. | Processes for obtaining microbial oil from microbial cells |
| US10392578B2 (en) | 2010-06-01 | 2019-08-27 | Dsm Ip Assets B.V. | Extraction of lipid from cells and products therefrom |
| US10472316B2 (en) | 2013-12-20 | 2019-11-12 | Dsm Ip Assets B.V. | Processes for obtaining microbial oil from microbial cells |
| US11124736B2 (en) | 2013-12-20 | 2021-09-21 | Dsm Ip Assets B.V. | Processes for obtaining microbial oil from microbial cells |
| CN117229848A (en) * | 2023-07-20 | 2023-12-15 | 青岛和合汇途工程技术有限公司 | Wet fractionation method for preparing 2-bit DHA algae oil |
-
2011
- 2011-11-08 CN CN 201110350200 patent/CN102388988B/en not_active Expired - Fee Related
Non-Patent Citations (2)
| Title |
|---|
| 何东平等: "微生物油脂精炼的研究", 《中国油脂》 * |
| 周林: "微生物油脂研究概况", 《粮食与食品工业》 * |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10329515B2 (en) | 2000-01-19 | 2019-06-25 | Dsm Ip Assets B.V. | Solventless extraction process |
| US10392578B2 (en) | 2010-06-01 | 2019-08-27 | Dsm Ip Assets B.V. | Extraction of lipid from cells and products therefrom |
| CN103421605B (en) * | 2012-05-25 | 2017-03-15 | 丰益(上海)生物技术研发中心有限公司 | Method and its application that oils and fatss are carried with erythritol fatty acid ester point |
| CN103421605A (en) * | 2012-05-25 | 2013-12-04 | 丰益(上海)生物技术研发中心有限公司 | Method for fractionating grease through using erythritol fatty acid ester, and application of erythritol fatty acid ester |
| US10342772B2 (en) | 2013-12-20 | 2019-07-09 | Dsm Ip Assets B.V. | Processes for obtaining microbial oil from microbial cells |
| US10364207B2 (en) | 2013-12-20 | 2019-07-30 | Dsm Ip Assets B.V. | Processes for obtaining microbial oil from microbial cells |
| US10472316B2 (en) | 2013-12-20 | 2019-11-12 | Dsm Ip Assets B.V. | Processes for obtaining microbial oil from microbial cells |
| TWI708836B (en) * | 2013-12-20 | 2020-11-01 | 荷蘭商Ｄｓｍ智慧財產有限公司 | Processes for obtaining microbial oil from microbial cells |
| US11124736B2 (en) | 2013-12-20 | 2021-09-21 | Dsm Ip Assets B.V. | Processes for obtaining microbial oil from microbial cells |
| CN105176671B (en) * | 2014-06-18 | 2018-11-30 | 中国石油化工股份有限公司 | A kind of method of the broken wall and auxiliary agent assisted extraction grease of oil-containing micro-algae |
| CN105176671A (en) * | 2014-06-18 | 2015-12-23 | 中国石油化工股份有限公司 | Method for wall breaking of oil-containing microalgae and assistant assisted extraction of oil |
| CN105454466A (en) * | 2015-12-04 | 2016-04-06 | 润科生物工程（福建）有限公司 | Application of sucrose fatty acid ester serving as grease crystallization inhibitor in oil containing polyunsaturated fatty acid single-cell grease |
| CN108624403A (en) * | 2018-04-28 | 2018-10-09 | 武汉工程大学 | A kind of method of low temperature non-organic solvent extraction energy insect oils |
| CN117229848A (en) * | 2023-07-20 | 2023-12-15 | 青岛和合汇途工程技术有限公司 | Wet fractionation method for preparing 2-bit DHA algae oil |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102388988B (en) | 2013-01-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102388988B (en) | Separated extraction method of microorganism oil | |
| CA3103242C (en) | Methods of refining a grain oil composition feedstock, and related systems, compositions and uses | |
| CN205501251U (en) | A simple and easy winterization device for fish oil | |
| US4870011A (en) | Method for obtaining lipids from fungus bodies | |
| WO2000036059A1 (en) | Two phase extraction of oil from biomass | |
| WO2012109642A1 (en) | Aqueous extraction methods for high lipid microorganisms | |
| CN102229860A (en) | Supersonic wave method for promoting rapid crystallization and fractionation of palm oil | |
| CN103131529B (en) | A kind of method extracting microbial oil | |
| CN104017645B (en) | The table crystalline substance copolymerization of a kind of peony seed oil is combined the method carrying out dewaxing with alcohol extraction | |
| CN103614239B (en) | Method for preparing low-temperature-resistant palm oil with high iodine value | |
| CN104531342B (en) | A kind of method for extracting microbial grease | |
| CN107629873B (en) | Method for enriching fish oil EPA and DHA through low-temperature crystallization | |
| CN104629906B (en) | A kind of crystallization in motion agent and point method for mentioning grease | |
| CN103467432B (en) | A kind of method extracting vitamin E from deodorizer distillate of idesia polycarpa oil | |
| CN103787864B (en) | Method for extracting DHA (Docosahexaenoic acid) from marine microalgae fermentation liquor | |
| CN105831370A (en) | Preparation method of cocoa butter equivalent | |
| CN108285826B (en) | Animal oil extraction method | |
| CN104293473A (en) | Method for extracting oil from oil production microbes | |
| US4272447A (en) | Crude edible oil wax removal process | |
| CN104293482A (en) | Rice bran fat oil, and preparation method and product thereof | |
| CN109528784B (en) | Method for extracting and preparing 4,4' -dimethyl sterol from shea butter | |
| CN104293474A (en) | Method for extracting oil from oil production microbes | |
| CN107083280B (en) | A kind of method using antimicrobial fiber separate microorganism grease | |
| CN102504962B (en) | Process for preparing polyunsaturated fatty acids (PUFAs) from microbial origin | |
| JP5947064B2 (en) | Method for producing fatty acid composition |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130123 |