WO2017039227A1 - Method for producing biodiesel and method for separating cells - Google Patents

Abstract

The present invention provides a method for producing biodiesel, the method comprising: (a) a step of feeding resuspended cultured microalgae cells into the entrance of a microchannel device having an ion-selective membrane so that the cells can pass through from the entrance to the exit of the device; (b) a step of forming an ion depletion zone by applying an electric field to the microchannel device and thereby causing ion concentration polarization (ICP) to occur in a predetermined region between the entrance and the exit; and (c) a step of separating oil contained in the microalgae cells from the microalgae cells on the basis of the ion depletion zone.

Description

Biodiesel production method and cell separation method

The present invention relates to a biodiesel production method and a cell separation method, and more particularly, a biodiesel production method and a cell capable of producing a large amount of biodiesel from microalgal cells and separating internal substances surrounded by cell membranes from cells. To a separation method.

Since fossil fuels that contributed to the development of human society are depleted resources with limited reserves, it is necessary to find sustainable energy resources. Biodiesel derived from living organisms is in the spotlight as the next renewable energy source because of its similar energy density to fossil fuels. Since microalgal cells, photosynthetic microorganisms, contain up to 70% of the lipid that is a raw material of biodiesel, the lipid production efficiency is improved through the upstream process of the culture method that increases the biomass per unit process and the downstream process of separating and extracting the lipids in the body. And research to improve the recovery rate is in progress.

However, the production cost of biodiesel exceeds that of petroleum due to the complicated steps in the extraction process. In particular, current technologies such as filtration, centrifugation, sedimentation, flotation, flocculation, etc. applied in downstream processes are economically important factors that determine 58% of the production cost, but are complex and time-consuming and expensive due to the nature of large-scale processes. do. In order to secure production economics, a method of extracting lipids through mass modularization of subunits is urgently needed.

The present invention is to solve a number of problems including the above problems, and the present invention relates to a biodiesel production method that is simple, economical, and mass-produced.

In addition, the present invention is to solve a number of problems, including the above problems, and to a cell separation method that can separate and extract the internal material by breaking the cell membrane of the cell. However, these problems are exemplary, and the scope of the present invention is not limited thereby.

According to one aspect of the invention, a biodiesel production method is provided. The biodiesel production method includes the steps of: (a) supplying resuspended microalgae cells to the inlet so as to pass from the inlet to the outlet of the microchannel device having an ion-selective membrane; (b) applying an electric field to the microchannel device to form an ion depletion zone by generating an ion concentration polarization (ICP) at a predetermined region between the inlet and the outlet; And (c) separating oil contained in the microalgal cells from the microalgal cells based on the ion depletion region.

In the biodiesel production method, an ion depletion zone is formed by generating an ion concentration polarization (ICP) at a portion adjacent to a branch point between the first outlet and the second outlet, The microalgal cells are separated into microalgal cell fragments and the oil based on the ion depletion region, and the separated microalgal cell fragments and the oil are pushed out by an electric repulsive force at the interface of the ion depletion region. May exit 2 outlets.

In the biodiesel production method, the microalgal cells may be crushed under shear stress in the vicinity of the ion depletion zone and separated into the microalgal cell fragments and the oil based on the ion depletion zone. have.

In the biodiesel production method, the microalgal cell fragments and the oil are micro-nano sized microparticles, and the microparticles having polarity may be subjected to dielectric polarization by an electric field.

In the biodiesel production method, the microalgal cells may be separated into the microalgal cell fragments and the oil by the force of the electroosmotic flow and the force of the ion concentration polarization phenomenon.

The biodiesel production method may further include separating and collecting the microalgae cells and the oil flowing out to the second outlet in a separate device.

In the biodiesel production method, the oil, hydrodynamic convection flow flowing out to the second outlet having a vector component in a direction parallel to at least one surface of the membrane and a vector component in a direction perpendicular to at least one surface of the membrane It may include.

In the biodiesel production method, the second outlet may have a predetermined angle with the first outlet and may be disposed at an obtuse angle with respect to the advancing direction of the fluid.

In the biodiesel production method, the ion selective membrane may use Nafion, and the ion selective membrane may be disposed to be electrically grounded at a branch point between the first outlet and the second outlet.

In the biodiesel production method, in the step (a), the resuspension culture of the microalgal cells, (1) culturing the microalgal cells in the first culture; (2) suspending and separating the microalgal cells by centrifugation; And (3) resuspending the microalgal cells suspended and separated in a second culture.

In the biodiesel production method, the first culture medium may include Tri-Acetate-Phosphate (TAP), and the second culture medium may include TAP-N.

In the biodiesel production method, the step of suspension separation of the microalgal cells of the step (2), the microalgal cells are separated from the first culture by the centrifugation method, suspended using distilled water It may comprise the step of separating.

In the biodiesel production method, in the step (3), the resuspension culture, resuspend culture of the microalgae cells for 5 to 7 days, body body to the resuspended microalgae cells After the treatment (BODIPY) may include the step of confirming that the oil particles are generated in the cells of the microalgal cells.

And, according to one aspect of the invention, there is provided a cell separation method. The cell separation method comprises: (a) a cell having a cell membrane and an inner material surrounded by the cell membrane at the inlet for passage from an inlet to an outlet of a microchannel device having an ion-selective membrane; Supplying; (b) applying an electric field to the microchannel device to form an ion depletion zone by generating an ion concentration polarization (ICP) at a predetermined region between the inlet and the outlet; And (c) separating the internal material from the cell based on the ion depletion region.

According to one embodiment of the present invention made as described above, it is possible to implement a method for producing biodiesel from microalgal cells with a simple structure and low power by using a microchannel device having an ion selective membrane. In addition, the cell membrane of the cells is crushed to have the effect of separating and extracting the internal substances. Of course, the scope of the present invention is not limited by these effects.

1A and 1B schematically illustrate a micro channel device according to an embodiment of the present invention.

FIG. 1C is a photograph of the micro channel device shown in FIG. 1A.

Figure 2 is a photograph of observing microalgal cells according to an experimental example of the present invention.

Figure 3 is a photograph analyzing the biodiesel separated by the biodiesel production method according to an experimental example of the present invention.

Figure 4 is a picture of analyzing the biodiesel coming along the separation channel of the micro-channel according to an experimental example of the present invention.

<Description of the code>

10: micro channel device

12: entrance

14: first exit

16: second exit

18: ion selective membrane

20: microalgal cells, cells

20a: Microalgae cell fragments, cell membrane fragments

20b: oil, substances inside the cell

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. However, the present invention is not limited to the embodiments disclosed below, but can be implemented in various forms, and the following embodiments are intended to complete the disclosure of the present invention, the scope of the invention to those skilled in the art It is provided to inform you completely. In addition, the components may be exaggerated or reduced in size in the drawings for convenience of description.

Embodiments of the present invention will now be described with reference to the drawings, which schematically illustrate ideal embodiments of the present invention. In the figures, for example, variations in the shape shown may be expected, depending on manufacturing techniques and / or tolerances. Accordingly, embodiments of the inventive concept should not be construed as limited to the specific shapes of the regions shown herein, but should include, for example, changes in shape resulting from manufacturing.

In general, in the extraction step of biodiesel, the conventional method of using compression is the simplest but the extraction efficiency is low. The supercritical fluid method shows extraction efficiency close to 100%, but there is a problem in that the economic efficiency is low due to the high initial installation and maintenance cost of the equipment. In addition, the microalgae may be lyophilized by a solvent extraction method and then extracted using a solvent such as chloroform and hexane. However, an economical extraction method is required because a solvent recovery process based on distillation and expensive solvents are used.

In addition, the process of drying the microalgae before extracting the biodiesel takes a drying cost and time, so the extraction method using the non-dried microalgae can improve the economics of the biodiesel, but the non-dried extraction method is very low to date It is showing efficiency. This is because microalgae have very high elastic modulus and water is a lubricant when they have moisture, making cell transformation difficult. As a result, extraction is also inefficient.

In order to solve this problem, the present invention provides a biodiesel production method capable of efficiently extracting biodiesel without drying the microalgae. The present invention also provides a cell separation method capable of separating the cell membrane and the inner material and extracting the inner material from the cell having the cell membrane and the inner material surrounded by the cell membrane.

A method of producing biodiesel in more detail below will be described with reference to FIGS. 1 to 4.

1 (a) and (b) schematically show a micro channel device according to an embodiment of the present invention, Figure 1 (c) is a photograph of the micro channel device shown in Figure 1 (a) to be.

Referring to FIG. 1A, the biodiesel production method may use a micro channel device 10 having an ion-selective membrane 18. The micro channel device 10 may include an ion selective membrane 18 within the micro channel device 10 to cause an ion concentration polarization phenomenon. The ion selective membrane 18 may be formed by specifying the space between the dielectric microparticles by collecting microalgal cells including biodiesel with each other. For example, microalgae cells are sheared near the ion selective membrane 18 and crushed into oil and crushed cell pieces. The oil and crushed cell pieces behave like charged particles due to dielectric polarization by an external electric field.

In addition, the ion concentration polarization phenomenon is one of the electrochemical transfer phenomena observed around the structure having a nano-membrane. It is theoretically known that when the thickness of the electric double layer is similar to the size of the nanomembrane, the double layer overlaps inside the nanomembrane to show single ion permeability. As the charges such as wall charges do not pass through the nanomembrane due to diffusion and drift force, only ions having opposite charges to the wall charge pass through, resulting in depletion and excess of ions at the nanomembrane interface. Among the ions that do not pass through the nano-membrane strong electrical repulsive force is acting affects both positive and negative ions, resulting in the ion concentration tool phenomenon. At this time, a vortex is formed around the boundary of the ion depletion region P, and charged particles, cells, and droplets are also pushed around the nano-membrane under the influence of the electrical repulsion of ions at the interface of the ion depletion region P. I will.

In (a) of FIG. 1, the microalgal cells are supplied from the left inlet 12 in the direction of the arrow to be crushed into oil and crushed microalgal cell fragments through the ion depletion region P, and the pulverized microalgae Cell fragments and oil may flow out of the second outlet 16. Thereafter, additional steps are required to collect the microalgal cell fragments and the oil spilled from a separate device, and separately collect the collected microalgal cell fragments and the oil. In addition, the microalgal cells are crushed into microalgal cell fragments and oil, and the process of removing the water contained in the microalgal cells when collecting the crushed microalgal cell fragments and oil is required. At the same time, the microalgal cells can be crushed and a part of the water can be removed, and the water removal rate can be controlled according to the channel size toward the first outlet 14 and the second outlet 16.

More specifically, the microalgal cells used in one embodiment of the present invention may be cultured through a pretreatment process. For example, microalgal cells may be cultured in a first culture. Here, the first culture medium may include, for example, Tri-Acetate-Phosphate (TAP). When the growth of the microalgae cells reaches an exponential phase in which cell division is actively performed, the microalgae cells in the culture medium can be separated by centrifugation and suspended by distilled water.

Thereafter, resuspended microalgal cells having a volume equal to that of the initial first culture and suspended in a second culture without a nitrogen source are resuspended. Here, the second culture may include, for example, TAP-N. In addition, the suspension separation process and the resuspension cultivation process is different only in the components of the culture medium, the culture method is the same.

Microalgal cells are resuspended in culture for about 5 to 7 days. After resuspended cultured microalgae cells were treated with lipid-specific fluorescence staining bodiphy (BODIPY), it was confirmed whether lipid microparticles with green fluorescence were generated in microalgae cells. The microalgal cells in which the lipid granules are generated can separate oil contained in the microalgal cells by using ion concentration polarization (ICP).

That is, the biodiesel production method according to an embodiment of the present invention is as follows. A micro-nano channel device 10 with an ion selective membrane 18 is fabricated. The micro channel device 10 may have at least two outlets. For example, the outlet may consist of a first outlet 14 and a second outlet 16. In this case, the second outlet 16 may be disposed in a direction inclined at an angle to the first outlet 14 at a predetermined angle and disposed at an obtuse angle with respect to the advancing direction of the fluid. The ion selective membrane 18 may be disposed in the vicinity of the branch point between the first outlet 14 and the second outlet 16.

The resuspended microalgae cells may be injected into the inlet 12 of the prepared micro nano-channel device 10. Resuspended microalgae cells are micro-nano sized microparticles, and polarized microparticles may be subjected to dielectric polarization by an electric field (E).

Referring to FIG. 1B again, the microalgal cells 20 are introduced into the microchannel device 10, and the electric field (2) is formed at both ends of the inlet 12 and the outlet 14, 16 of the microchannel device 10. E) can be applied. Ion Concentration Polarization (ICP) occurs in a region adjacent to the branch point between the first outlet 14 and the second outlet 16 by the applied electric field E, thereby causing an ion depletion zone, P) can be formed. The microalgal cells 20 may be separated into microalgal cell fragments 20a and oil 20b based on the ion depletion region P. At this time, the remains may be pushed out at the interface of the ion depletion region P by the force due to the electroosmotic flow and the force due to the ion concentration polarization phenomenon.

On the other hand, after making a micro-nano channel coupling structure, if the material to be separated is put on the top of the micro channel and a voltage is applied to both ends of the nano channel, an ion concentration polarization phenomenon due to the ion concentration gradient occurs. Microalgae cells are crushed under shear stress τ = Q / A (Q is shear force and A is area) from the vortex generated at the ion depletion layer interface, and the oil contained in the cells is released in the aqueous solution. Crushed cell fragments and oil droplets are dielectrically polarized by an external electric field (E), behaving like charged particles, and are pushed away from the nanomembrane by electrostatic forces at the edge of the ion depletion layer caused by ion concentration polarization. Will receive strength. Force from the fluid flow F _drag = -6πμ = α (Ｕ is the flow velocity, μ is the viscosity of the fluid and α is the representative radius of the particle), and it is the force due to the ion concentration polarization phenomenon F _ICP and the force from the electroosmotic flow It is pushed in the opposite direction to the nanomembrane by F _EOF .

In other words, when an electric field is applied to the microchannel and the membrane 18, the reverse direction of the membrane 18 at the inlet of the membrane 18 due to the difference between the flow rate of the microchannel and the flow through the membrane 18 is reversed. A flow occurs and a vortex occurs accordingly. The vortex exerts a shear stress on the surface of the microalgal cells.

Therefore, the separated microalgal cell fragments 20a and oil 20b are pushed out by the electric repulsive force at the interface of the ion depletion region P and flow out to the second outlet 16. The oil 20b has a fluid flowing out to the second outlet 16 having a vector component parallel to at least one surface of the ion selective membrane 18 and a vector component perpendicular to at least one surface of the ion selective membrane 18. Mechanical convective flow. The oil 20b flowing out to the second outlet 16 can be collected in a separate device. When the oil 20b is collected, a solution having a high proportion of oil can be obtained.

In addition, referring to FIG. 1C, the micro channel device 10 may use a transparent material as a first substrate. For example, one of pyrex, silicon dioxide, silicon nitride, quartz or SU-8 may be used as the first substrate. The micro channel device 10 is coated with a low-autofluorescent material. In addition to the first substrate, a second substrate may be included. The second substrate can be used to cover or seal the micro channel device 10. The second substrate may be made of the same material as the first substrate. In some embodiments the first substrate and the second substrate may be made of different materials.

Meanwhile, the manufacture of the micro channel device 10 can be completed by plasma bonding the first substrate to the second substrate. The substrate is a support structure of the micro channel device 10. At least a portion of the substrate may be made of silicon. In one embodiment of the invention, the substrate, device or portions of the device may be made of a polymer. The polymer may be polydimethylsiloxane (PDMS). In the case of using PDMS, oxygen (O 2) plasma may be treated to have hydrophilicity, but in some cases, oxygen plasma treatment may be omitted.

In addition, the micro channel device 10 includes an inlet 12 through which microalgae flow, and a first outlet 14 and a second outlet 16 may be formed on opposite sides. An ion selective membrane 18 may be disposed at an area adjacent to the branch point between the first outlet 14 and the second outlet 16 to be electrically grounded (GND). By the ion selective membrane 18, the microalgal cell oil 20b and the ground microalgal cell fragment 20a introduced through the inlet 12 may be discharged to the second outlet 16. Here, the ion selective membrane 18 may use, for example, Nafion.

In addition, the ion selective membrane 18 may be predominantly behaving against cations that do not match the ionic conductivity in the electrolyte. As a result, an ion concentration gradient can be generated on both sides of the ion selective membrane 18. Once ion concentration polarization is induced near the cation exchange ion selective membrane 18, both the cation and anion concentrations decrease on the anode side and increase on the cathode side of the junction. Moreover, charged particles, cells, other small colloids, and the like, may similarly exhibit ion depletion or ionic hyperplasia, which makes it possible to obtain a depletion region in rectified state.

Hereinafter, an experimental example to which the above-described technical concept is applied will be described to help understanding of the present invention. However, the following experimental examples are only for helping understanding of the present invention, and the present invention is not limited to the following experimental examples.

As a sample according to the experimental example of the present invention, microalgae culture was performed in TAP (Tris-Acetate-Phosphate) medium. When the growth of the microalgae reached the exponential stage in which cell division actively occurs, the microalgae in the culture medium were separated by centrifugation and then suspended in distilled water. Resuspension cultures were performed in TAP-N medium without a nitrogen source at the same volume as the initial culture. After 5-7 days, after treatment with BODIPY, a lipid-specific fluorescent dye, it was confirmed whether liposomes with green fluorescence were generated in the cells.

Thereafter, the resuspended microalgae were placed on top of the microchannel and voltage was applied across the nanochannel. At this time, the ion concentration polarization phenomenon occurs due to the ion concentration gradient, and the oil contained in the microalgae is separated and concentrated separately at the outlet of the microchannel by the ion concentration polarization phenomenon.

Figure 2 is a photograph of observing microalgal cells according to an experimental example of the present invention.

Figure 2 (a) is a microscopic analysis of microalgae cells in white light, Figure 2 (b) is a microscopic analysis of the case of blue light applied to the microalgae cells fluorescently stained lipid.

Referring to Figure 2, it can be seen that microalgal cells are well cultured and suspended in TAP medium and TAP-N. In addition, blue-light was applied to the resuspended microalgae cells after treatment with lipid-specific fluorescent dye bodily (BODIPY). You can see that this is well generated.

Figure 3 is a photograph analyzing the biodiesel separated by the biodiesel production method according to an experimental example of the present invention.

(A), (b) and (c) of FIG. 3 are photographs of oil cells being torn by shear stress when the ion concentration polarization (ICP) phenomenon occurs under white light. 3 (d), 3 (e) and 3 (f) are photographs of cells crushed by shear stress and extracting oil grains when an ion concentration polarization (ICP) phenomenon occurs in a state in which blue light is reflected.

Referring to FIGS. 1A and 3, in both cases, cell fragments crushed by the ion depletion region P formed near the branch point between the first outlet 14 and the second outlet 16. Fine algae cells 20a and oil 20b granules separated into 20a and oil 20b and pushed to the opposite side of the ion-selective membrane 18 by electrostatic repulsion and crushed to the second outlet 16 You can see that they are leaking.

Figure 4 is a picture of analyzing the biodiesel coming along the separation channel of the micro-channel according to an experimental example of the present invention.

Referring to FIG. 4, the photograph shows observation of oil grains coming out of the separation channel of the micro channel device 10. It can be seen that the oil (20b) can be separated and concentrated by grinding the microalgae.

As described above, the present invention can separate and concentrate oil contained in microalgae cells based on a low-cost device based on PDMS having a micro-nano channel binding system. The nonpolar cells, which have advanced only from the conventionally charged materials, are also affected by ion concentration polarization when microscopic formations occur.

In addition, the ion depletion region may be formed to have a different size depending on the size of the external electric field applied to the micro channel device. In one embodiment of the present invention, the ion depletion region is formed to have a size substantially equal to the cross-sectional area of the microchannel, so that the oil separated from the microalgal cells can all flow out to the second outlet by electrical repulsive force. However, if it is applied smaller than the size of the electric field shown in one embodiment of the present invention, some of the oil may be moved along the interface of the ion depletion region, so that at least a part of the oil may flow out to the first outlet.

Therefore, the cross-sectional area of the channel of the micro-channel device should be designed in a condition similar to that of the electric double layer, or the value of the electric field should be appropriately selected and applied so that the size of the ion depletion region fills the cross-sectional area of the channel.

Meanwhile, the optimization of industrial processes is essential for the optimized production of biodiesel. The present invention is a method for extracting lipids through a large-scale modularization of subunits, using micro-cell devices of low cost, to extract crushed and biodiesel using ion concentration polarization of microalgal cells containing biodiesel. Tracking and analyzing the entire process of lipid production in the device, and building industrial facilities based on this data, is very valuable because it can extract large amounts of biodiesel.

The biodiesel production method described above with reference to FIGS. 1 to 4 can be applied not only to microalgal cells but also to separation of other cells. Such cells should be understood to include microalgae, erythrocytes, as well as any constructs that are surrounded by cell membranes and have internal material therein.

That is, the cell separation method according to an embodiment of the present invention is as follows. A micro-nano channel device 10 with an ion selective membrane 18 is fabricated. The micro channel device 10 may have at least two outlets. For example, the outlet may consist of a first outlet 14 and a second outlet 16. In this case, the second outlet 16 may be disposed in a direction inclined at an angle to the first outlet 14 at a predetermined angle and disposed at an obtuse angle with respect to the advancing direction of the fluid. The ion selective membrane 18 may be disposed in the vicinity of the branch point between the first outlet 14 and the second outlet 16.

The inlet 12 of the prepared micro nano-channel device 10 may be injected with a cell 20 having an inner material 20b surrounded by the cell membrane 20a and the cell membrane 20a such as red blood cells or the like. The cells 20 are micro-nano sized particles, and the polarized particles can be subjected to dielectric polarization by the electric field (E).

The cell 20 may be introduced into the micro channel device 10, and an electric field E may be applied to both the inlet 12 and the outlets 14 and 16 of the micro channel device 10. Ion Concentration Polarization (ICP) occurs in a region adjacent to the branch point between the first outlet 14 and the second outlet 16 by the applied electric field E, thereby causing an ion depletion zone, P) can be formed. The cell 20 may be separated into cell membrane fragments 20a and an internal material (contents inside the cell membrane) 20b based on the ion depletion region P. At this time, the remains may be pushed out at the interface of the ion depletion region P by the force due to the electroosmotic flow and the force due to the ion concentration polarization phenomenon.

In other words, when an electric field is applied to the microchannel and the membrane 18, the reverse direction of the membrane 18 at the inlet of the membrane 18 due to the difference between the flow rate of the microchannel and the flow through the membrane 18 is reversed. A flow occurs and a vortex occurs accordingly. The vortex exerts a shear stress on the cell membrane 20a.

Therefore, the cell membrane 20a is crushed by the shear stress, and the separated cell membrane fragments 20a and the inner substance (contents inside the cell membrane) 20b are pushed out by the electric repulsive force at the interface of the ion depletion region P. 2 exits the outlet 16. The inner material 20b flows out to the second outlet 16 while having a vector component parallel to at least one surface of the ion selective membrane 18 and a vector component perpendicular to at least one surface of the ion selective membrane 18. Hydrodynamic convective flow. The internal material 20b flowing out to the second outlet 16 can be collected in a separate device. In the collected solution, a high proportion of the internal material 20b may be formed.

Although the present invention has been described with reference to one embodiment shown in the drawings, this is merely exemplary, and those skilled in the art will understand that various modifications and equivalent other embodiments are possible therefrom. Therefore, the true technical protection scope of the present invention will be defined by the technical spirit of the appended claims.

Claims (14)

(a) feeding resuspended microalgal cells to the inlet for passage from the inlet to the outlet of the microchannel device having an ion-selective membrane; (b) applying an electric field to the microchannel device to form an ion depletion zone by generating an ion concentration polarization (ICP) at a predetermined region between the inlet and the outlet; And (c) separating oil contained in the microalgal cells from the microalgal cells based on the ion depletion region; Including, Biodiesel production method. The method of claim 1, The outlet branches to the first outlet and the second outlet, and ion concentration polarization (ICP) occurs at a portion adjacent to the branch point between the first outlet and the second outlet, thereby causing ion depletion. zones are formed, The microalgal cells are separated into microalgal cell fragments and the oil based on the ion depletion region, and the separated microalgal cell fragments and the oil are pushed out by an electric repulsive force at the interface of the ion depletion region. 2, which flows out of the outlet Biodiesel production method. The method of claim 1, The microalgal cells are crushed under shear stress in the vicinity of the ion depletion zone and separated into the microalgal cell fragments and the oil based on the ion depletion zone. Biodiesel production method. The method of claim 1, The microalgae pieces and the oil are micro-nano sized particles, and the polarized particles can be subjected to dielectric polarization by an electric field. Biodiesel production method. The method of claim 1, The microalgal cells are separated into the microalgal cell fragments and the oil by the force of the electroosmotic flow and the force of the ion concentration polarization phenomenon, Biodiesel production method. The method of claim 2, And separating and collecting the microalgal cells and the oil flowing out to the second outlet in a separate device. Biodiesel production method. The method of claim 2, The oil comprises a hydrodynamic convection flow exiting the second outlet having a vector component in a direction parallel to at least one surface of the membrane and a vector component in a direction perpendicular to at least one surface of the membrane, Biodiesel production method. The method of claim 2, The second outlet has a predetermined angle with the first outlet and is a direction disposed at an obtuse angle with respect to the advancing direction of the fluid; Biodiesel production method. The method of claim 2, The ion selective membrane uses Nafion, The ion selective membrane is arranged to be electrically grounded at a branch point between the first outlet and the second outlet, Biodiesel production method. The method of claim 1, In the step (a), the resuspension culture of the microalgal cells, (1) culturing the microalgal cells in the first culture medium; (2) suspending and separating the microalgal cells by centrifugation; And (3) resuspending the microalgal cells suspended and separated in a second culture; Performed through, Biodiesel production method. The method of claim 10, The first culture solution contains Tri-Acetate-Phosphate (TAP), The second culture solution comprises TAP-N, Biodiesel production method. The method of claim 10, In the step (2), the step of suspending the microalgal cells, Separating the microalgal cells from the first culture medium by the centrifugation method, and the suspension separation using distilled water, Biodiesel production method. The method of claim 10, In step (3), the step of resuspending culture, After resuspending the microalgae cells for 5 to 7 days, and performing body bodily (BODIPY) treatment on the resuspended microalgae cells to determine whether the oil particles are generated in the cells of the microalgae cells. Comprising the steps of: Biodiesel production method. (a) supplying a cell having a cell membrane and an inner material surrounded by the cell membrane to the inlet for passage from an inlet to an outlet of a microchannel device having an ion-selective membrane; (b) applying an electric field to the microchannel device to form an ion depletion zone by generating an ion concentration polarization (ICP) at a predetermined region between the inlet and the outlet; And (c) separating the internal material from the cell based on the ion depletion region; Including, Cell separation method.

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