CN106967491A - A kind of algae oil highly effective extraction method of rich oil Chaetoceros - Google Patents
A kind of algae oil highly effective extraction method of rich oil Chaetoceros Download PDFInfo
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- CN106967491A CN106967491A CN201710152287.3A CN201710152287A CN106967491A CN 106967491 A CN106967491 A CN 106967491A CN 201710152287 A CN201710152287 A CN 201710152287A CN 106967491 A CN106967491 A CN 106967491A
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
- C11B1/04—Pretreatment of vegetable raw material
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/10—Production of fats or fatty oils from raw materials by extracting
- C11B1/104—Production of fats or fatty oils from raw materials by extracting using super critical gases or vapours
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Plant Substances (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of algae oil highly effective extraction method of rich oil Chaetoceros, by centrifuging, being collected by filtration by the mixed liquor obtained after Chaetoceros culture, expanded algal gel is then obtained using expelling-expansion pretreatment, digested, is dried, then using supercritical CO2Demulsification extraction is carried out, ultrafiltration membrance filter obtains algae oil.The method of the present invention carries out multiple extraction to the part containing algae oil in Chaetoceros, significantly improves extraction efficiency.The algae oil highly effective extraction method of described rich oil Chaetoceros has very important application prospect in terms of production of biodiesel.
Description
Technical field
The invention belongs to the algae oil extractive technique field of rich oil Chaetoceros, specifically it is related to a kind of rich oil Chaetoceros
Algae oil highly effective extraction method.
Background technology
In recent years, as fossil fuel is increasingly reduced, reproducible biodiesel receives much concern.Biodiesel it is basic into
It is divided into fatty acid ester, mainly triglycerides or free fatty is reacted with lower alcohols by transesterification reaction and obtained.In early days
Raw material for biodiesel synthesis for the oil crops such as soybean, rape, sunflower seeds, corn but these edible oil costs too
It is high.Also there is the vegetable oil after using waste cooking oils such as animal fat or cooking later for raw material, but be limited to supply,
It is unable to large-scale application.At present, various countries are generally using unedible oil material such as palm, manioca, cocoa chocolate tree, cottonseed, False flax
Crop, the large-scale tree crop such as rich grease-contained Chinese pistache, hat fruit, hair Lai trees is also wide concerned within the border for China.However, woody work
Thing is slow-growing, and biotransformation efficiency is low,
Cause holding at high price for current biodiesel, it is difficult to popularization and application.Therefore, oil content is high, biomass is big, easily
In the new lover of culture and the high oil-rich microalgae of biotransformation efficiency as researcher, also it is regarded as most possibly substituting fossil combustion
The renewable oil resource of material, reclaims in the excavation of algae germ plasm resource, training systern, ripe algae and biodiesel turns at present
The links such as change become study hotspot.
Microalgae is single celled planktonic algae, can be grown in fresh water or salt water, with good resistance, at present generally
Cultivated on marginal soil, production grain arable land will not be taken.The growth of microalgae is very fast, and one times of biomass can be lifted daily,
Doubling time during exponential phase of growth can be shortened to less than 4h, and microalgae annual production is than current production rate highest oil plant under unit area
Taller 7~23 times of crop.Early stage people concern microalgae is because a variety of microalgaes contain substantial amounts of unrighted acid, including two
Dodecahexaene acid (Docosahex-aenoic acid, DHA) and eicosapentaenoic acid (Eicosapen-taenoic
Acid, EPA), it is respectively provided with high economic value.Found that the chain saturated fatty acids synthesized by some microalgaes were occupied later big absolutely
Ratio, is the excellent raw material of biodiesel synthesis, is also had a good application prospect in terms of bioenergy.
Therefore, the efficient purification for oil-rich microalgae just seems significant, not only has in biodiesel application aspect
Good application prospect, the content and composition for analyzing its aliphatic acid is effective using with theory directive significance to its.Chaetoceros are rich
Containing long chain fatty acids, there is good application value in the green energy resource industry such as biodiesel.
The content of the invention
Present invention solves the technical problem that a kind of algae oil highly effective extraction method of rich oil Chaetoceros is there is provided, from rich oil angle
The artificial fast culture of hair algae arrives algae oil high efficiency extraction again, and design cycle is rationally, simple to operate, and recovery rate is high.
The technical scheme is that：A kind of algae oil highly effective extraction method of rich oil Chaetoceros, is mainly included the following steps that：
Step one：It is prepared by Chaetoceros culture medium：CO(NH2)2(urea) 10-20mg, KH2PO45-10mg, FeC6H5O7·
5H2O 0.4-0.8mg, EDTA 3-5mg, Na2SiO3·9H2O 10-12mg, NaHCO3300-500mg, 920 plant growths
Stimulin 3-5 international units, VB10.1-0.3mg, VB60.1-0.3mg, VB120.05-0.09mg, ferric citrate 1-2mg,
ZnSO4300-500mg, volume ratio 2-4% molybdic acid aqueous solution 5-15ml, manganese sulfate 5-8mg, potassium dihydrogen phosphate 5-15mg, chlorine
Change and described culture medium is prepared after potassium 3-5mg, the seawater 1000mL after sterilization, mixed dissolution.
Step 2：The artificial fast culture of Chaetoceros：Chaetoceros algae kind is selected, it is miscellaneous without other foreign peoples by microexamination
Algae, protozoan etc., Chaetoceros algae kind is accessed in the culture vessel equipped with culture medium, and inoculum density is 2-3 × 105cell/
ML, seals culture vessel and starts to carry out illumination in air-charging incubation, incubation, light illumination is 3000-4000lx；By 4-
The culture of 6 days, density is up to 4 × 106-5×106Cell/mL is to obtain can be used to the Chaetoceros mixed liquor for extracting algae oil, and culture is extremely
Stationary phase is grown, the intracellular of algae kind gradually accumulates grease.
Step 3：Algae oil high efficiency extraction：Chaetoceros mixed liquor is centrifuged in the presence of rotating speed is 4000-6000r/min
10-20min, is collected by filtration algae substances, then expanded algal gel is obtained using expelling-expansion pretreatment, by expanded material and 3-5
Times amount water be mixed to get mixed liquor, into mixed liquor add 0.02 times of volume alkali protease digested, after enzymolysis from
The isolated free oil of the heart, emulsion, hydrolyzate and residue, adjust pH value to be neutrality emulsion, and moisture is 3- after drying
5%, 80 mesh are pulverized and sieved, then using supercritical CO2Carry out demulsification extraction, then using 0.22 μm of ultrafiltration membrance filter after, i.e.,
Obtain algae oil.
Further, the selection standard of described Chaetoceros algae kind is that well-grown, viability be vigorous, bright in colour, nothing
Precipitate and without obvious attached wall phenomenon.
Further, described air-charging incubation refers to：Air and carbon dioxide mix are filled with 10-12L/min speed
Gas, volume ratio ratio is VAir∶VCarbon dioxide=8: 1, night stops inflation, and between-line spacing inflation, that is, inflated 1 hour, stopped 1 small daytime
When, repeat.
Further, described super critical condition is the 16- that entrainment reagent ethanol addition is emulsion quality after drying
20%, CO2Flow 6-10L/min, extracting pressure 30-50MPa, extraction time 1-3h, 35-45 DEG C of extraction temperature.
Further, described culture vessel is the plastic containers of cylinder, and body diameter is 50-70cm, is highly
100-120cm, can be sealed, and have the breather pipe with switch in the middle of lid, and breather pipe connects aerating system, and described is logical
Gas system is respectively divided into air control system for air and carbon dioxide control system, is connected respectively the ventilation of air and carbon dioxide
Pipe, is annular multi-tube structure, and fumarole, which reaches deep down into container, can reduce liquid and be detained on annular spread pipe, inner air tube
Flow velocity takes in 20m/s or so, aerating system and is provided with concentration sensor, so as to preferably control the concentration and volume of gas, bottom
There is the Drainage pipe that screen pack is connected in portion.
Further, described culture vessel disinfection way sterilizes for potassium permanganate.
Further, described seawater sterilization method is that will pass through the bolting silk net mistake of 300 mesh of seawater of precipitation and husky filter
Filter, then to every 1m3300ml available chlorine contents 8-12% liquor natrii hypochloritis is added in seawater, contains effective chlorine in seawater
Amount is reached after 15-20mg/L, sterilization 24h, and sodium thiosulfate 30g is added wherein, to neutralize chlorine residue, that is, completes seawater sterilization.
Compared with prior art, beneficial effects of the present invention are embodied in：The mixed liquor obtained after Chaetoceros culture is centrifuged,
It is collected by filtration, expanded algal gel is then obtained using expelling-expansion pretreatment, digested, dries, then using supercritical CO2Enter
Row demulsification extraction, ultrafiltration membrance filter obtains algae oil.The method of the present invention carries out bring up again more to the part containing algae oil in Chaetoceros
Take, significantly improve extraction efficiency.
Embodiment
For ease of the understanding of the present invention, below in conjunction with explanation is further explained exemplified by specific embodiment, implement
Example does not constitute the restriction to the embodiment of the present invention.
Embodiment 1：A kind of algae oil highly effective extraction method of rich oil Chaetoceros, is mainly included the following steps that：
Step one：It is prepared by Chaetoceros culture medium：CO(NH2)2(urea) 10mg, KH2PO45mg, FeC6H5O7·5H2O
0.4mg, EDTA 3mg, Na2SiO3·9H2O 10mg, NaHCO3300mg, the international unit of 920 plant growth substance 3, VB1
0.1mg, VB60.1mg, VB120.05mg, ferric citrate 1mg, ZnSO4300mg, the molybdic acid aqueous solution of volume ratio 2%
Described in being prepared after 5ml, manganese sulfate 5mg, potassium dihydrogen phosphate 5mg, potassium chloride 3mg, the seawater 1000mL after sterilization, mixed dissolution
Culture medium.
Step 2：The artificial fast culture of Chaetoceros：Chaetoceros algae kind is selected, it is miscellaneous without other foreign peoples by microexamination
Algae, protozoan etc., Chaetoceros algae kind is accessed in the culture vessel equipped with culture medium, and inoculum density is 2 × 105Cell/mL,
Sealing culture vessel simultaneously starts to carry out illumination in air-charging incubation, incubation, and light illumination is 3000lx；By the culture of 4 days,
Density is up to 4 × 106Cell/mL is to obtain can be used to the Chaetoceros mixed liquor for extracting algae oil, and culture extremely grows stationary phase, algae kind
It is intracellular gradually to accumulate grease.
Step 3：Algae oil high efficiency extraction：Chaetoceros mixed liquor is centrifuged into 10min in the presence of rotating speed is 4000r/min,
Algae substances are collected by filtration, expanded algal gel is then obtained using expelling-expansion pretreatment, expanded material is mixed with the water of 3 times of amounts
Mixed liquor is obtained, the alkali protease that 0.02 times of volume is added into mixed liquor is digested, and trip is centrifugally separating to obtain after enzymolysis
From oil, emulsion, hydrolyzate and residue, adjust pH value to be neutrality emulsion, moisture is 3% after drying, pulverize and sieve 80
Mesh, then using supercritical CO2Carry out demulsification extraction, then using 0.22 μm of ultrafiltration membrance filter after, that is, obtain algae oil.
Wherein, the selection standard of described Chaetoceros algae kind be well-grown, viability it is vigorous, bright in colour, without precipitation
And without obvious attached wall phenomenon.Described air-charging incubation refers to：Air and carbon dioxide mix gas are filled with 10L/min speed
Body, volume ratio ratio is VAir∶VCarbon dioxide=8: 1, night stops inflation, and between-line spacing inflation, that is, inflated 1 hour, stopped 1 small daytime
When, repeat.Described super critical condition is 16%, the CO that entrainment reagent ethanol addition is emulsion quality after drying2Stream
Measure 6L/min, extracting pressure 30MPa, extraction time 1h, 35 DEG C of extraction temperature.Described culture vessel holds for the plastics of cylinder
Device, body diameter is 50cm, is highly 100cm, can seal, and has the breather pipe with switch, breather pipe in the middle of lid
Aerating system is connected, the Drainage pipe that screen pack is connected is arranged at bottom.Described culture vessel disinfection way sterilizes for potassium permanganate.
Described seawater sterilization method is, by the bolting silk net filtration by precipitation and 300 mesh of seawater of husky filter, then to every 1m3Sea
The liquor natrii hypochloritis of 300ml available chlorine contents 8% is added in water, available chlorine content in seawater is reached 15mg/L, sterilizes 24h
Afterwards, sodium thiosulfate 30g is added wherein, to neutralize chlorine residue, that is, completes seawater sterilization.
Embodiment 2：A kind of algae oil highly effective extraction method of rich oil Chaetoceros, is mainly included the following steps that：
Step one：It is prepared by Chaetoceros culture medium：CO(NH2)2(urea) 15mg, KH2PO47.5mg, FeC6H5O7·5H2O
0.6mg, EDTA 4mg, Na2SiO3·9H2O 11mg, NaHCO3400mg, the international unit of 920 plant growth substance 4, VB1
0.2mg, VB60.2mg, VB120.07mg, ferric citrate 1.5mg, ZnSO4400mg, the molybdic acid aqueous solution of volume ratio 3%
Institute is prepared after 10ml, manganese sulfate 6.5mg, potassium dihydrogen phosphate 10mg, potassium chloride 4mg, the seawater 1000mL after sterilization, mixed dissolution
The culture medium stated.
Step 2：The artificial fast culture of Chaetoceros：Chaetoceros algae kind is selected, it is miscellaneous without other foreign peoples by microexamination
Algae, protozoan etc., Chaetoceros algae kind is accessed in the culture vessel equipped with culture medium, and inoculum density is 2.5 × 105cell/
ML, seals culture vessel and starts to carry out illumination in air-charging incubation, incubation, light illumination is 3500lx；By the training of 5 days
Support, density is up to 4.5 × 106Cell/mL is to obtain can be used to the Chaetoceros mixed liquor for extracting algae oil, and culture extremely grows stationary phase,
The intracellular of algae kind gradually accumulates grease.
Step 3：Algae oil high efficiency extraction：Chaetoceros mixed liquor is centrifuged into 15min in the presence of rotating speed is 5000r/min,
Algae substances are collected by filtration, expanded algal gel is then obtained using expelling-expansion pretreatment, expanded material is mixed with the water of 4 times of amounts
Mixed liquor is obtained, the alkali protease that 0.02 times of volume is added into mixed liquor is digested, and trip is centrifugally separating to obtain after enzymolysis
From oil, emulsion, hydrolyzate and residue, adjust pH value to be neutrality emulsion, moisture is 4% after drying, pulverize and sieve 80
Mesh, then using supercritical CO2Carry out demulsification extraction, then using 0.22 μm of ultrafiltration membrance filter after, that is, obtain algae oil.
Wherein, the selection standard of described Chaetoceros algae kind be well-grown, viability it is vigorous, bright in colour, without precipitation
And without obvious attached wall phenomenon.Described air-charging incubation refers to：Air and carbon dioxide mix gas are filled with 11L/min speed
Body, volume ratio ratio is VAir∶VCarbon dioxide=8: 1, night stops inflation, and between-line spacing inflation, that is, inflated 1 hour, stopped 1 small daytime
When, repeat.Described super critical condition is 18%, the CO that entrainment reagent ethanol addition is emulsion quality after drying2Stream
Measure 8L/min, extracting pressure 40MPa, extraction time 2h, 40 DEG C of extraction temperature.Described culture vessel holds for the plastics of cylinder
Device, body diameter is 60cm, is highly 110cm, can seal, and has the breather pipe with switch, breather pipe in the middle of lid
Aerating system is connected, the Drainage pipe that screen pack is connected is arranged at bottom.Described culture vessel disinfection way sterilizes for potassium permanganate.
Described seawater sterilization method is, by the bolting silk net filtration by precipitation and 300 mesh of seawater of husky filter, then to every 1m3Sea
The liquor natrii hypochloritis of 300ml available chlorine contents 10% is added in water, available chlorine content in seawater is reached 17.5mg/L, is sterilized
After 24h, sodium thiosulfate 30g is added wherein, to neutralize chlorine residue, that is, completes seawater sterilization.
Embodiment 3：A kind of algae oil highly effective extraction method of rich oil Chaetoceros, is mainly included the following steps that：
Step one：It is prepared by Chaetoceros culture medium：CO(NH2)2(urea) 20mg, KH2PO410mg, FeC6H5O7·5H2O
0.8mg, EDTA 5mg, Na2SiO3·9H2O 12mg, NaHCO3500mg, the international unit of 920 plant growth substance 5, VB1
0.3mg, VB60.3mg, VB120.09mg, ferric citrate 2mg, ZnSO4500mg, the molybdic acid aqueous solution of volume ratio 4%
Prepared after 15ml, manganese sulfate 8mg, potassium dihydrogen phosphate 15mg, potassium chloride 5mg, the seawater 1000mL after sterilization, mixed dissolution described
Culture medium.
Step 2：The artificial fast culture of Chaetoceros：Chaetoceros algae kind is selected, it is miscellaneous without other foreign peoples by microexamination
Algae, protozoan etc., Chaetoceros algae kind is accessed in the culture vessel equipped with culture medium, and inoculum density is 3 × 105Cell/mL,
Sealing culture vessel simultaneously starts to carry out illumination in air-charging incubation, incubation, and light illumination is 4000lx；By the culture of 6 days,
Density is up to 5 × 106Cell/mL is to obtain can be used to the Chaetoceros mixed liquor for extracting algae oil, and culture extremely grows stationary phase, algae kind
It is intracellular gradually to accumulate grease.
Step 3：Algae oil high efficiency extraction：Chaetoceros mixed liquor is centrifuged into 20min in the presence of rotating speed is 6000r/min,
Algae substances are collected by filtration, expanded algal gel is then obtained using expelling-expansion pretreatment, expanded material is mixed with the water of 5 times of amounts
Mixed liquor is obtained, the alkali protease that 0.02 times of volume is added into mixed liquor is digested, and trip is centrifugally separating to obtain after enzymolysis
From oil, emulsion, hydrolyzate and residue, adjust pH value to be neutrality emulsion, moisture is 5% after drying, pulverize and sieve 80
Mesh, then using supercritical CO2Carry out demulsification extraction, then using 0.22 μm of ultrafiltration membrance filter after, that is, obtain algae oil.
Wherein, the selection standard of described Chaetoceros algae kind be well-grown, viability it is vigorous, bright in colour, without precipitation
And without obvious attached wall phenomenon.Described air-charging incubation refers to：Air and carbon dioxide mix gas are filled with 12L/min speed
Body, volume ratio ratio is VAir∶VCarbon dioxide=8: 1, night stops inflation, and between-line spacing inflation, that is, inflated 1 hour, stopped 1 small daytime
When, repeat.Described super critical condition is 20%, the CO that entrainment reagent ethanol addition is emulsion quality after drying2Stream
Measure 10L/min, extracting pressure 50MPa, extraction time 3h, 45 DEG C of extraction temperature.Described culture vessel is cylindrical plastics
Container, body diameter is 70cm, is highly 120cm, can seal, and has the breather pipe with switch, ventilation in the middle of lid
Pipe connects aerating system, and the Drainage pipe that screen pack is connected is arranged at bottom.Described culture vessel disinfection way disappears for potassium permanganate
Poison.Described seawater sterilization method is, by the bolting silk net filtration by precipitation and 300 mesh of seawater of husky filter, then to every 1m3
The liquor natrii hypochloritis of 300ml available chlorine contents 12% is added in seawater, available chlorine content in seawater is reached 20mg/L, is sterilized
After 24h, sodium thiosulfate 30g is added wherein, to neutralize chlorine residue, that is, completes seawater sterilization.
Embodiment 4：Isolated rich oil Chaetoceros are cultivated from Bay in Shenzhen mangrove seawater, then by implementing
The extracting method of example 2 is extracted, and then its lubricant component is detected by testing.
1. taking the algae oil after 1mL filterings, it is placed in gas-chromatography sample cell, using gas chromatography-mass spectrometry instrument
(gas chromatograph is Trace1300, and mass spectrograph is TraceISQ, Thermo Fisher, USA) is analyzed.Chromatographic column is
DB-5ms capillary columns (35m × 0.25 μm of 250 μ m), scanning charge-mass ratio scope is 40~500.Carrier gas is He, flow velocity 1mL
min-1, the μ L of sample size 1；280 DEG C of injector temperature.Elevated Temperature Conditions are：70 DEG C of 2min of initial temperature, then with 10 DEG C of min-1
220 DEG C are warming up to, then with 3 DEG C of min-1260 DEG C are warming up to, then with 10 DEG C of min-1280 DEG C are warming up to, 5min is kept.
With 37 kinds of mixed methyl aliphatic ester standard items (NU-CHEK-PREP, INC., USA) for reference.By comparing standard sample and counting
Peak area is calculated, the content of each component is determined.
2. being analyzed through GC-MS, mass spectrogram is compared with fatty acid methyl ester standard items, determine institute's fatty acids in algae oil into
Point.Gas phase collection of illustrative plates shows that fatty acid species are mainly the long chain fatty acids of 14~18 carbon atoms in rich oil Chaetoceros algae oil, are gone out
Peak time is concentrated between the 15th minute to 20 minutes, as shown in figure 1, using 37 kinds of fatty acid methyl ester hybrid standard product as reference,
And compared through mass spectral database, it is respectively C14: 0, C15: 0, C16: 1, C16: 0, C18: 1 and C18: 0, institute to determine the peak 1~6 in figure
Obtain algae oil Analysis of Fatty Acids Composition table such as following table (mgmL-1)：
| C14 | C15 | C16 | C18 | It is total | |
| Component | 11.62 | 2.14 | 36.54 | 1.94 | 52.24 |
3. be can be seen that with reference to data above by the algae oil constituent analysis to the Chaetoceros, in algae oil mainly into
It is divided into the major class Short-Chain Fatty Acids of C14: 0 and C16 two, this Fatty acid compositions make this kind of algae oil be very suitable for producing biological bavin
Oil.Therefore the algae oil highly effective extraction method of described rich oil Chaetoceros has very important application in terms of production of biodiesel
Prospect.
Finally illustrate：It is that, for the ease of understanding embodiments of the invention, the present invention can also have using above-mentioned technical proposal
Other embodiment, protection scope of the present invention is not limited to this.Without departing from the spirit and substance of the case in the present invention, affiliated skill
The technical staff in art field works as can make various corresponding changes and deformation according to the present invention, but these corresponding changes and deformation
Belong to the scope of the claims of the present invention.
Claims (7)
1. a kind of algae oil highly effective extraction method of rich oil Chaetoceros, it is characterised in that mainly include the following steps that：
Step one：It is prepared by Chaetoceros culture medium：CO(NH2)2(urea) 10-20mg, KH2PO45-10mg, FeC6H5O7·5H2O
0.4-0.8mg, EDTA 3-5mg, Na2SiO3·9H2O 10-12mg, NaHCO3300-500mg, 920 plant growth stimulatings
Plain 3-5 international units, VB10.1-0.3mg, VB60.1-0.3mg, VB120.05-0.09mg, ferric citrate 1-2mg,
ZnSO4300-500mg, volume ratio 2-4% molybdic acid aqueous solution 5-15ml, manganese sulfate 5-8mg, potassium dihydrogen phosphate 5-15mg, chlorine
Change and described culture medium is prepared after potassium 3-5mg, the seawater 1000mL after sterilization, mixed dissolution.
Step 2：The artificial fast culture of Chaetoceros：Select Chaetoceros algae kind, by microexamination without the miscellaneous algae of other foreign peoples,
Protozoan etc., Chaetoceros algae kind is accessed in the culture vessel equipped with culture medium, and inoculum density is 2-3 × 105Cell/mL,
Sealing culture vessel simultaneously starts to carry out illumination in air-charging incubation, incubation, and light illumination is 3000-4000lx；By 4-6 days
Culture, density is up to 4 × 106-5×106Cell/mL is to obtain can be used to the Chaetoceros mixed liquor for extracting algae oil.
Step 3：Algae oil high efficiency extraction：Chaetoceros mixed liquor is centrifuged into 10- in the presence of rotating speed is 4000-6000r/min
20min, is collected by filtration algae substances, then obtains expanded algal gel using expelling-expansion pretreatment, expanded material is measured with 3-5 times
Water be mixed to get mixed liquor, into mixed liquor add 0.02 times of volume alkali protease digested, after enzymolysis centrifugation divide
From free oil, emulsion, hydrolyzate and residue is obtained, pH value is adjusted to be neutrality emulsion, moisture is 3-5% after drying,
80 mesh are pulverized and sieved, then using supercritical CO2Carry out demulsification extraction, then using 0.22 μm of ultrafiltration membrance filter after, that is, obtain
Algae oil.
2. a kind of algae oil highly effective extraction method of rich oil Chaetoceros as claimed in claim 1, it is characterised in that described angle hair
The selection standard of algae algae kind is well-grown, viability is vigorous, bright in colour, without precipitation and without obvious attached wall phenomenon.
3. a kind of algae oil highly effective extraction method of rich oil Chaetoceros as claimed in claim 1, it is characterised in that described inflation
Culture refers to：Air and carbon dioxide gas mixture are filled with 10-12L/min speed, volume ratio ratio is VAir∶VCarbon dioxide=
8: 1, night stops inflation, and between-line spacing inflation, that is, inflated 1 hour, stopped 1 hour, repeat daytime.
4. a kind of algae oil highly effective extraction method of rich oil Chaetoceros as claimed in claim 1, it is characterised in that described super to face
Boundary's condition is the 16-20%, CO2 flow 6-10L/min, extracting pressure that entrainment reagent ethanol addition is emulsion quality after drying
30-50MPa, extraction time 1-3h, 35-45 DEG C of extraction temperature.
5. a kind of algae oil highly effective extraction method of rich oil Chaetoceros as claimed in claim 1, it is characterised in that described culture
Container is the plastic containers of cylinder, and body diameter is 50-70cm, is highly 100-120cm, can seal, in lid
Between have with switch breather pipe, breather pipe connection aerating system, bottom have screen pack connect Drainage pipe.
6. a kind of algae oil highly effective extraction method of rich oil Chaetoceros as claimed in claim 1, it is characterised in that described culture
Container disinfection way sterilizes for potassium permanganate.
7. a kind of algae oil highly effective extraction method of rich oil Chaetoceros as claimed in claim 1, it is characterised in that described seawater
Sterilization method is, by the bolting silk net filtration by precipitation and 300 mesh of seawater of husky filter, then to every 1m3Added in seawater
300ml available chlorine contents 8-12% liquor natrii hypochloritis, makes available chlorine content in seawater reach 15-20mg/L, sterilizes 24h
Afterwards, sodium thiosulfate 30g is added wherein, to neutralize chlorine residue, that is, completes seawater sterilization.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710152287.3A CN106967491A (en) | 2017-03-15 | 2017-03-15 | A kind of algae oil highly effective extraction method of rich oil Chaetoceros |
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