CN108129420B - Method for extracting fucoxanthin from phaeodactylum tricornutum by dimethyl ether fluid - Google Patents
Method for extracting fucoxanthin from phaeodactylum tricornutum by dimethyl ether fluid Download PDFInfo
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Abstract
The invention discloses a method for extracting fucoxanthin from phaeodactylum tricornutum by dimethyl ether fluid, which comprises the following steps: 1) freeze thawing and wall breaking of fresh Phaeodactylum tricornutum algae; 2) performing dynamic gradient countercurrent extraction on the wall-broken algae mud by using dimethyl ether fluid; 3) the fucoxanthin is separated from the extract liquid by exsolution; the method can effectively avoid the damage of physical and chemical dehydration to the effective components in the microalgae, greatly reduce the energy consumption in the extraction process, perform dynamic gradient countercurrent extraction operation, effectively improve the concentration of the effective components in the extraction liquid, reduce the dosage of an extracting agent, improve the extraction efficiency, realize the high-efficiency extraction of the phaeodactylum tricornutum fucoxanthin, and have reliable safety.
Description
Technical Field
The invention belongs to the technical field of microalgae cultivation, and particularly relates to a method for extracting fucoxanthin from phaeodactylum tricornutum by using dimethyl ether fluid.
Background
Fucoxanthin, also called algin and fucoxanthin, molecular formula C42H58O6The structural formula is as follows, the molecular weight is 658.91, the density is 1.09, the melting point is 166-168 ℃, the CAS (CAS registry for standards) number is 3351-86-8, the allene carotenoid belongs to, the carotenoid is an important carotenoid contained in brown algae, diatom, chrysophyceae and yellow green algae, the brown algae are brown substances, and the index pigment of diatom plants is also used.
The fucoxanthin has a unique structure, and has potential value advantages with application prospects in various pharmacological activities, wherein the physiological activities of resisting inflammation, tumors, obesity, oxidation, pox, diabetes, malaria and blood fat are partially proved, and other potential physiological activities are actively researched and explored by scientists.
At present, a plurality of raw materials can be used for extracting fucoxanthin, such as brown algae, diatom and other algae. The Phaeodactylum tricornutum is also a typical unicellular diatom, and the Phaeodactylum tricornutum has the characteristics of easy culture, perennial growth, high growth speed and easy regulation, and has greater advantages in producing fucoxanthin by culturing the Phaeodactylum tricornutum.
The traditional method for extracting fucoxanthin from Phaeodactylum tricornutum is an organic solvent extraction method, the method is simple and easy to implement, and the more common organic solvents comprise methanol, ethanol, acetone, diethyl ether, petroleum ether, dimethyl sulfoxide and the like, and the organic solvents can be used independently or mixed according to a certain proportion. However, the organic solvent extraction method has obvious limitations such as high extraction temperature, fucoxanthin loss, product solvent residue, potential safety hazard and the like.
Many researchers have been working on exploring new extraction techniques such as supercritical extraction, pressurized solvent extraction, microwave extraction, ultrasonic extraction, etc. The novel extraction methods have multiple advantages, and can improve extraction efficiency, shorten extraction time, save extraction cost, and reduce harm to environment. However, the use of microwaves and ultrasound is merely an aid and needs to be combined with other extraction techniques to function. The supercritical extraction technology has also been studied in the extraction of fucoxanthin, but the supercritical solvent CO is a supercritical solvent2Extraction processing can be carried out only in an ultrahigh pressure state of more than 25MPa, and the industrial application of the extraction is greatly restricted. The pressurized solvent extraction method is a rapid and effective extraction method for extracting bioactive substances by using an organic solvent at higher temperature and pressure, and the method greatly reduces the dosage of the organic solvent, but fails to change the technical defects of the organic solvent extraction method.
The subcritical extraction method is a novel extraction and separation technology developed in the last decade. The subcritical extraction method is a novel extraction and separation technology which utilizes subcritical fluid as an extractant, transfers fat-soluble components in solid materials into a liquid extractant through molecular diffusion of the extraction materials and the extractant in a soaking process according to the principle that organic matters are similar and compatible in a closed, oxygen-free and low-pressure container, and separates the extractant from a target product through a reduced pressure evaporation process to finally obtain the target product. Subcritical fluid extraction has many advantages over other separation methods: the method has the advantages of no toxicity, no harm, no pollution, no hot processing, no damage to active products of the retained extract, no oxidation, high yield, energy conservation, low operation cost and easy separation from the products, and can be used for industrial large-scale production.
Patent CN 101077991a discloses a method for extracting natural pyrethrin by subcritical dimethyl ether fluid. Patent CN 10297006a discloses chrysanthemum absolute oil extracted by subcritical fluid low-temperature process. Patent CN104738251A discloses a method for subcritical fluid extraction of tea extract. Patent CN105400696A discloses a method for extracting zanthoxylum oil resin by subcritical dimethyl ether fluid. The four patents all adopt subcritical fluid dimethyl ether as an extracting agent, the process method is quite the same and different, only the materials are treated differently, but the extraction of the phaeodactylum tricornutum fucoxanthin is not involved.
In the production practice of fucoxanthin extraction from Phaeodactylum tricornutum, the water content of fresh Phaeodactylum tricornutum algae is usually over 60 percent, and the Phaeodactylum tricornutum algae fucoxanthin is greatly damaged by adopting various drying and dehydrating methods such as thermal processing, electromagnetic processing, vacuum processing and the like; the siliceous outer shell of the phaeodactylum tricornutum is combined tightly, organic solvent is difficult to permeate into cytoplasm effectively to contact with liposome such as pigment, and the extraction efficiency of fucoxanthin is low; the fucoxanthin content of the phaeodactylum tricornutum is about 1.0 percent, the concentration of the extraction liquid is quite low, and the extraction solvent and the energy consumption are high. The common subcritical extraction method adopts static immersion extraction which is difficult to meet the requirement.
Because the material characteristics are unique, the active ingredients have active properties, and higher requirements are necessarily provided for the extraction of fucoxanthin from phaeodactylum tricornutum.
Disclosure of Invention
The invention provides a method for extracting fucoxanthin from phaeodactylum tricornutum by dimethyl ether fluid at low temperature, high efficiency and safety aiming at the process defects and shortcomings in the existing technology for extracting the fucoxanthin from the phaeodactylum tricornutum.
In order to solve the technical problems, the technical scheme of the invention is as follows:
a method for extracting fucoxanthin from Phaeodactylum tricornutum by dimethyl ether fluid comprises the following steps:
1) freeze thawing and wall breaking of fresh Phaeodactylum tricornutum algae;
2) performing dynamic gradient countercurrent extraction on the wall-broken algae mud by using dimethyl ether fluid;
3) and (4) carrying out desolventizing on the extract liquor to separate out fucoxanthin.
As a preferable technical scheme, the Phaeodactylum tricornutum fresh algae in the step 1) is fresh Phaeodactylum tricornutum algae which is dried by a high-speed centrifuge, and the water content is 65.0-75.0% by mass.
As a preferred technical scheme, the phaeodactylum tricornutum fresh algae is quickly frozen, then refrigerated and finally unfrozen to realize freeze-thaw wall breaking, wherein the quick freezing temperature is-70 ℃ to-40 ℃, the quick freezing time is 8.0h to 12.0h, the refrigerated temperature is-20 ℃ to-10 ℃, the unfreezing temperature is 20 ℃ to 30 ℃, the unfreezing time is 0.5 to 1.5h, and the freeze-thaw times are 1 to 3 times.
As a preferable technical scheme, the dimethyl ether fluid in the step 2) is subcritical fluid, the pressure is 0.5MPa to 1.0MPa, and the temperature is 40 ℃ to 60 ℃.
As a preferable technical scheme, the residence time of the dimethyl ether fluid in the step 2) is 1.0-2.0 hours
As a preferred technical scheme, the extraction operation in the step 2) is dynamic gradient countercurrent extraction, the dimethyl ether fluid sequentially and continuously passes through a group of extraction tanks, the concentration of dimethyl ether in a single tank is gradually increased, a single tank at the front end of the extraction operation is separated from an extraction system, a single tank filled with wall-broken algae mud is connected to the tail end of the extraction system, and the dimethyl ether fluid and the wall-broken algae mud reversely flow.
As a preferred technical scheme, the extraction tank group is formed by connecting 4-6 single tanks in series.
As a preferable technical scheme, the pressure of the exsolution of the extraction liquid in the step 3) is 0.05MPa to 0.10MPa, and the temperature is 50 ℃ to 70 ℃.
As a preferable technical scheme, fucoxanthin is produced after oil-water centrifugal separation of the desolventized crude fucoxanthin, and the rotating speed of a centrifugal machine is 4500-10000 r/min.
Adopt the produced beneficial effect of above-mentioned technical scheme to lie in:
1) the wet algae mud is mechanically dehydrated, and directly enters a processing system after external water is removed, so that the damage of physical and chemical dehydration to effective components in the microalgae can be effectively avoided, and the energy consumption in the extraction process is greatly reduced.
2) The freeze thawing wall breaking and fresh algae collecting and storing process complement each other, on one hand, because the high saline water carried by the fresh algae has strong permeability, the saline water enters into cells to form ice crystals and expand to destroy cell walls, on the other hand, the fresh algae needs to be frozen to reduce water activity and collected and stored.
3) The dimethyl ether is soluble in water, the solubility of the dimethyl ether in every 100g of water is up to 30g at room temperature, the dimethyl ether fluid is greatly promoted to permeate into the phaeodactylum tricornutum cells to extract pigments and lipids such as fat-soluble fucoxanthin, and the dimethyl ether fluid is in a subcritical state, the extraction temperature is low, and the active ingredients of the phaeodactylum tricornutum are basically not damaged.
4) The dynamic gradient countercurrent extraction operation can effectively improve the concentration of the effective components of the extraction liquid, reduce the dosage of the extractant, improve the extraction efficiency and realize the high-efficiency extraction of the fucoxanthin of the phaeodactylum tricornutum.
5) Dimethyl ether is gas at normal temperature and normal pressure, can be thoroughly removed from fucoxanthin, and the extraction process is always controlled at low temperature, is basically a physical dissolution and evaporation process, and the fucoxanthin keeps the natural characteristics thereof, thereby having reliable safety.
Due to the adoption of the technical scheme, the method for extracting fucoxanthin from phaeodactylum tricornutum by using the dimethyl ether fluid comprises the following steps: 1) freeze thawing and wall breaking of fresh Phaeodactylum tricornutum algae; 2) performing dynamic gradient countercurrent extraction on the wall-broken algae mud by using dimethyl ether fluid; 3) the fucoxanthin is separated from the extract liquid by exsolution; the method can effectively avoid the damage of physical and chemical dehydration to the effective components in the microalgae, greatly reduce the energy consumption in the extraction process, perform dynamic gradient countercurrent extraction operation, effectively improve the concentration of the effective components in the extraction liquid, reduce the dosage of an extracting agent, improve the extraction efficiency, realize the high-efficiency extraction of the phaeodactylum tricornutum fucoxanthin, and have reliable safety.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a diagram comparing freeze-thaw wall-broken cell states of Phaeodactylum tricornutum of the present invention;
FIG. 2 is a high performance liquid chromatogram of fucoxanthin according to the present invention.
Detailed Description
A method for extracting fucoxanthin from Phaeodactylum tricornutum by dimethyl ether fluid comprises the following steps:
1) freeze thawing and wall breaking of fresh Phaeodactylum tricornutum algae;
2) performing dynamic gradient countercurrent extraction on the wall-broken algae mud by using dimethyl ether fluid;
3) and (4) carrying out desolventizing on the extract liquor to separate out fucoxanthin.
The Phaeodactylum tricornutum fresh algae in the step 1) is fresh algae which is obtained by spin-drying Phaeodactylum tricornutum through a high-speed centrifuge, and the water content is 65.0-75.0% by mass.
The phaeodactylum tricornutum fresh algae is quickly frozen, then refrigerated and finally unfrozen to realize freeze-thaw wall breaking, the quick freezing temperature is-70 ℃ to-40 ℃, the quick freezing time is 8.0h to 12.0h, the refrigerating temperature is-20 ℃ to-10 ℃, the unfreezing temperature is 20 ℃ to 30 ℃, the unfreezing time is 0.5 to 1.5h, and the number of times of freeze thawing is 1 to 3.
The dimethyl ether fluid in the step 2) is subcritical fluid, the pressure is 0.5MPa to 1.0MPa, and the temperature is 40 ℃ to 60 ℃.
The residence time of the dimethyl ether fluid in the step 2) is 1.0-2.0 hours
The extraction operation in the step 2) is dynamic gradient countercurrent extraction, the dimethyl ether fluid sequentially and continuously passes through a group of extraction tanks, the concentration of dimethyl ether in a single tank is gradually increased, the single tank at the front end of the extraction operation is separated from the extraction system, the single tank filled with the wall-broken algae mud is connected to the tail end of the extraction system, and the dimethyl ether fluid and the wall-broken algae mud reversely flow.
The extraction tank group is composed of 4-6 single tanks connected in series.
The pressure of desolventizing the extract liquor in the step 3) is 0.05MPa to 0.10MPa, and the temperature is 50 ℃ to 70 ℃.
And carrying out oil-water centrifugal separation on the desolventized crude fucoxanthin to produce fucoxanthin, wherein the rotating speed of a centrifugal machine is 4500-10000 r/min.
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The first embodiment is as follows:
a method for extracting fucoxanthin from Phaeodactylum tricornutum by dimethyl ether fluid comprises the following steps:
1) freeze thawing and wall breaking of fresh Phaeodactylum tricornutum algae;
2) performing dynamic gradient countercurrent extraction on the wall-broken algae mud by using dimethyl ether fluid;
3) and (4) carrying out desolventizing on the extract liquor to separate out fucoxanthin.
The Phaeodactylum tricornutum fresh algae in the step 1) is fresh algae which is obtained by spin-drying Phaeodactylum tricornutum through a high-speed centrifuge, and the water content is 65.0 percent by mass.
The Phaeodactylum tricornutum fresh algae is quickly frozen, then refrigerated and finally unfrozen to realize freeze-thaw wall breaking, wherein the quick freezing temperature is-70 ℃, the quick freezing time is 8.0h, the refrigerating temperature is-20 ℃, the unfreezing temperature is 20 ℃, the unfreezing time is 0.5, and the freeze-thaw times are 1.
The dimethyl ether fluid in the step 2) is subcritical fluid, the pressure is 0.5MPa, and the temperature is 40 ℃.
The residence time of the dimethyl ether fluid in the step 2) is 1.0 hour
The extraction operation in the step 2) is dynamic gradient countercurrent extraction, the dimethyl ether fluid sequentially and continuously passes through a group of extraction tanks, the concentration of dimethyl ether in a single tank is gradually increased, the single tank at the front end of the extraction operation is separated from the extraction system, the single tank filled with the wall-broken algae mud is connected to the tail end of the extraction system, and the dimethyl ether fluid and the wall-broken algae mud reversely flow.
The extraction tank group is composed of 4 single tanks connected in series.
The desolventizing pressure of the extraction liquid in the step 3) is 0.05MPa, and the temperature is 50 ℃.
And carrying out oil-water centrifugal separation on the desolventized crude fucoxanthin to produce fucoxanthin, wherein the rotating speed of a centrifugal machine is 4500 rpm.
Example two:
a method for extracting fucoxanthin from Phaeodactylum tricornutum by dimethyl ether fluid comprises the following steps:
1) freeze thawing and wall breaking of fresh Phaeodactylum tricornutum algae;
2) performing dynamic gradient countercurrent extraction on the wall-broken algae mud by using dimethyl ether fluid;
3) and (4) carrying out desolventizing on the extract liquor to separate out fucoxanthin.
The Phaeodactylum tricornutum fresh algae in the step 1) is fresh algae which is obtained by spin-drying Phaeodactylum tricornutum through a high-speed centrifuge, and the water content is 70.0 percent by mass.
The Phaeodactylum tricornutum fresh algae is quickly frozen, then refrigerated and finally unfrozen to realize freeze-thaw wall breaking, wherein the quick freezing temperature is-55 ℃, the quick freezing time is 102.0h, the refrigerating temperature is-15 ℃, the unfreezing temperature is 25 ℃, the unfreezing time is 1.0h, and the number of freeze-thaw times is 23.
The dimethyl ether fluid in the step 2) is subcritical fluid, the pressure is 0.8MPa, and the temperature is 50 ℃.
The residence time of the dimethyl ether fluid in the step 2) is 1.5 hours
The extraction operation in the step 2) is dynamic gradient countercurrent extraction, the dimethyl ether fluid sequentially and continuously passes through a group of extraction tanks, the concentration of dimethyl ether in a single tank is gradually increased, the single tank at the front end of the extraction operation is separated from the extraction system, the single tank filled with the wall-broken algae mud is connected to the tail end of the extraction system, and the dimethyl ether fluid and the wall-broken algae mud reversely flow.
The extraction tank group is composed of 5 single tanks connected in series.
The pressure of desolventizing the extract liquor in the step 3) is 0.08MPa, and the temperature is 60 ℃.
And carrying out oil-water centrifugal separation on the desolventized crude fucoxanthin to produce fucoxanthin, wherein the rotating speed of a centrifugal machine is 8000 revolutions per minute.
Example three:
a method for extracting fucoxanthin from Phaeodactylum tricornutum by dimethyl ether fluid comprises the following steps:
1) freeze thawing and wall breaking of fresh Phaeodactylum tricornutum algae;
2) performing dynamic gradient countercurrent extraction on the wall-broken algae mud by using dimethyl ether fluid;
3) and (4) carrying out desolventizing on the extract liquor to separate out fucoxanthin.
The Phaeodactylum tricornutum fresh algae in the step 1) is fresh algae which is obtained by spin-drying Phaeodactylum tricornutum through a high-speed centrifuge, and the water content is 5.0 percent by mass.
The Phaeodactylum tricornutum fresh algae is quickly frozen, then refrigerated and finally unfrozen to realize freeze-thaw wall breaking, wherein the quick freezing temperature is-40 ℃, the quick freezing time is 12.0h, the refrigerating temperature is-10 ℃, the unfreezing temperature is 30 ℃, the unfreezing time is 1.5h, and the freeze-thaw times are 3 times.
The dimethyl ether fluid in the step 2) is subcritical fluid, the pressure is 1.0MPa, and the temperature is 60 ℃.
The residence time of the dimethyl ether fluid in the step 2) is 2.0 hours
The extraction operation in the step 2) is dynamic gradient countercurrent extraction, the dimethyl ether fluid sequentially and continuously passes through a group of extraction tanks, the concentration of dimethyl ether in a single tank is gradually increased, the single tank at the front end of the extraction operation is separated from the extraction system, the single tank filled with the wall-broken algae mud is connected to the tail end of the extraction system, and the dimethyl ether fluid and the wall-broken algae mud reversely flow.
The extraction tank group is composed of 6 single tanks connected in series.
The desolventizing pressure of the extraction liquid in the step 3) is 0.10MPa, and the temperature is 70 ℃.
And carrying out oil-water centrifugal separation on the desolventized crude fucoxanthin to produce fucoxanthin, wherein the rotating speed of a centrifugal machine is 10000 revolutions per minute.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (6)
1. A method for extracting fucoxanthin from Phaeodactylum tricornutum by dimethyl ether fluid is characterized by comprising the following steps:
1) freeze thawing and wall breaking of the phaeodactylum tricornutum fresh algae, wherein the phaeodactylum tricornutum fresh algae in the step 1) is fresh algae obtained by spin-drying phaeodactylum tricornutum through a high-speed centrifuge, and the water content is 65.0-75.0% by mass; quick-freezing fresh Phaeodactylum tricornutum algae, refrigerating, and thawing to realize freeze-thaw wall breaking, wherein the quick-freezing temperature is-70 ℃ to-40 ℃, the quick-freezing time is 8.0h to 12.0h, the refrigerating temperature is-20 ℃ to-10 ℃, the thawing temperature is 20 ℃ to 30 ℃, the thawing time is 0.5 to 1.5h, and the number of freeze-thaw times is 1 to 3;
2) performing dynamic gradient countercurrent extraction on the wall-broken algae mud by using dimethyl ether fluid; the extraction operation is dynamic gradient countercurrent extraction, dimethyl ether fluid sequentially and continuously passes through a group of extraction tanks, the concentration of dimethyl ether in a single tank is gradually increased, the single tank at the front end of the extraction operation is separated from an extraction system, a new single tank filled with wall-broken algae mud is connected to the tail end of the extraction system, and the dimethyl ether fluid and the wall-broken algae mud reversely flow;
3) and (4) carrying out desolventizing on the extract liquor to separate out fucoxanthin.
2. The method for extracting fucoxanthin from phaeodactylum tricornutum with dimethyl ether fluid as claimed in claim 1, wherein: the dimethyl ether fluid in the step 2) is subcritical fluid, the pressure is 0.5MPa to 1.0MPa, and the temperature is 40 ℃ to 60 ℃.
3. The method for extracting fucoxanthin from phaeodactylum tricornutum with dimethyl ether fluid as set forth in claim 2, wherein: the residence time of the dimethyl ether fluid in the step 2) is 1.0-2.0 hours.
4. The method for extracting fucoxanthin from phaeodactylum tricornutum with dimethyl ether fluid as claimed in claim 1, wherein: the extraction tank group is composed of 4-6 single tanks connected in series.
5. The method for extracting fucoxanthin from phaeodactylum tricornutum with dimethyl ether fluid as claimed in claim 1, wherein: the pressure of desolventizing the extract liquor in the step 3) is 0.05MPa to 0.10MPa, and the temperature is 50 ℃ to 70 ℃.
6. The method for extracting fucoxanthin from Phaeodactylum tricornutum with dimethyl ether fluid as claimed in claim 5, wherein: and carrying out oil-water centrifugal separation on the desolventized crude fucoxanthin to produce fucoxanthin, wherein the rotating speed of a centrifugal machine is 4500-10000 r/min.
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