Particle analysis method and particle manufacturing method

Abstract

To provide a particle analysis method capable of analyzing whether or not a component to be included is included in a particle to be analyzed.SOLUTION: A particle analysis method comprises: a first magnetic susceptibility measuring step S4 of measuring the bulk magnetic susceptibility of a first particle p1; an inclusion processing step S5 of performing inclusion processing of including a component pt to be included smaller than the first particle p1 in the first particle p1; a second magnetic susceptibility measuring step S8 of measuring the bulk magnetic susceptibility of a second particle p2 to be analyzed being the first particle p1 after the inclusion processing; and a step S9 of analyzing whether a component pt to be included is included in the second particle p2 on the basis of the result measured by the first magnetic susceptibility measuring step S4 and the result measured by the second magnetic susceptibility measuring step S8.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a particle analysis method and a particle production method. [Background technology]

[0002] A microcapsule is a particle that encapsulates a liquid or solid component. The particle diameter of a microcapsule is smaller than 1 mm. A microcapsule has a membrane (capsule base material) that covers the encapsulated component. For example, various techniques have been proposed for industrially producing microcapsules that encapsulate active ingredients such as medicines, pesticides, fragrances, or food materials. Methods for manufacturing microcapsules include, for example, chemical methods, physical methods, and biological methods. Chemical methods include, for example, interfacial polymerization, liquid-based effective coating methods, and molecular inclusion methods using cyclodextrin. Physical methods include, for example, spray drying, spray cooling, and air suspension coating methods. Biological methods include, for example, a method of encapsulating components in the cytoskeleton of yeast or the like.

[0003] In the biological method, yeast-derived capsule materials are mainly used. The yeast-derived capsule materials are made of the yeast cytoskeleton remaining after removing soluble contents from the yeast. The cytoskeleton includes cell walls, cell membranes, and other solid components. Specifically, the yeast-derived capsule materials are made of the solid components of the yeast remaining after removing soluble components from the yeast cells by washing and enzyme treatments. The cytoskeleton of the yeast cell wall or the like is used as the capsule material. For example, Patent Document 1 discloses a technology for releasing intracellular components outside the yeast cells by enzyme treatment of the yeast.

[0004] Patent Document 1 discloses that a hydrophobic or hydrophilic substance is encapsulated in a capsule base material derived from yeast. Patent Document 1 discloses that the hydrophobic substance is at least one type of derived lipid selected from the group consisting of long-chain hydrocarbons, long-chain alcohols, long-chain amino alcohols, long-chain aldehydes, long-chain ketones, long-chain acids and their salts, terpenoids, steroids, and carotenoids. It also discloses that the hydrophilic substance is at least one type of substance selected from the group consisting of amino acids, water-soluble vitamins, nucleotides, and sugars.

[0005] Furthermore, Patent Document 1 discloses that the yeast is at least one type of microorganism selected from the group consisting of Saccharomyces cerevisiae, Saccharomyces rouxii, Saccharomyces carlsbergensisi, Candida utilis, Candida tropicalis, Candida lipolytica, and Candida flaveri.

[0006] Incidentally, in order to quantitatively analyze whether or not the component to be encapsulated is encapsulated in the microcapsule, for example, high performance liquid chromatography (HPLC) is generally used. Specifically, the encapsulated component is extracted from the microcapsule, the encapsulated component is quantified using a high performance liquid chromatograph, and the amount of the encapsulated component per 1 g of the microcapsule is calculated based on the measurement result, thereby analyzing (evaluating) whether or not the component to be encapsulated is encapsulated in the microcapsule. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Japanese Patent Application Publication No. 8-243378 Summary of the Invention [Problem to be solved by the invention]

[0008] However, quantitative analysis using high performance liquid chromatography merely estimates whether or not the target component is encapsulated in the microcapsules (the particles to be analyzed).

[0009] The present invention has been made in consideration of the above-mentioned problems, and an object of the present invention is to provide a particle analysis method and a particle manufacturing method that can analyze, for each analysis target, whether or not the particle to be analyzed contains a component to be contained therein. [Means for solving the problem]

[0010] The particle analysis method of the present invention includes a first observation step of magnetically migrating a first particle and observing it, a first migration velocity measurement step of measuring the magnetophoretic velocity of the first particle from the result of observation in the first observation step, a first magnetic susceptibility measurement step of measuring the volume magnetic susceptibility of the first particle based on the result of measurement in the first migration velocity measurement step, an inclusion processing step of performing an inclusion processing to cause the first particle to include a component to be included that is smaller than the first particle, a second observation step of magnetically migrating a second particle to be analyzed, which is the first particle after the inclusion processing, and observing it, a second migration velocity measurement step of measuring the magnetophoretic velocity of the second particle from the result of observation in the second observation step, a second magnetic susceptibility measurement step of measuring the volume magnetic susceptibility of the second particle based on the result of measurement in the second migration velocity measurement step, and a step of analyzing whether the component to be included is included in the second particle based on the result of measurement in the first magnetic susceptibility measurement step and the result of measurement in the second magnetic susceptibility measurement step.

[0011] In one embodiment, the particle analysis method further includes a step of spectroscopically observing the second particles to analyze whether or not the component to be included is included in the second particles.

[0012] In one embodiment, the particle analysis method further includes a first zeta potential measurement step of measuring the zeta potential of the first particle, a second zeta potential measurement step of measuring the zeta potential of the second particle, and a step of analyzing whether the component to be encapsulated is encapsulated in the second particle based on a result of the measurement in the first zeta potential measurement step and a result of the measurement in the second zeta potential measurement step.

[0013] The particle production method of the present invention includes the above-mentioned particle analysis method.

[0014] In one embodiment, the first particles contain solid components of the yeast that remain after intracellular components are released from the yeast.

[0015] In one embodiment, the particle production method further includes a first particle production step of producing the first particles. The first particle production step includes a step of releasing the intracellular components from the yeast.

[0016] In one embodiment, the encapsulation step includes a step of mixing the first particles and the component to be encapsulated in a solvent. Effect of the Invention

[0017] According to the particle analysis method and particle production method of the present invention, it is possible to analyze for each analyte whether or not the particle to be analyzed contains a component to be contained therein. [Brief description of the drawings]

[0018] [Figure 1] 1 is a flowchart showing a particle manufacturing method according to a first embodiment of the present invention. [Diagram 2] FIG. 1 is a schematic diagram of an analysis device according to a first embodiment of the present invention. [Diagram 3] 1A and 1B are diagrams illustrating the movement of particles. [Figure 4] FIG. 1 is a diagram showing a configuration of an analysis device according to a first embodiment of the present invention. [Diagram 5]5 is a flowchart showing a particle manufacturing method according to a second embodiment of the present invention. [Figure 6] 10 is a flowchart showing a particle manufacturing method according to a third embodiment of the present invention. [Figure 7] FIG. 4 is a diagram showing measurement results according to Example 1 of the present invention. [Figure 8] FIG. 11 is a diagram showing measurement results according to Example 2 of the present invention. [Figure 9] FIG. 11 is a diagram showing measurement results according to Example 3 of the present invention. [Figure 10] FIG. 11 is a diagram showing a first measurement result according to Example 4 of the present invention. [Figure 11] FIG. 11 is a diagram showing a second measurement result according to Example 4 of the present invention. [Figure 12] FIG. 11 is a diagram showing a third measurement result according to Example 4 of the present invention. [Figure 13] FIG. 13 is a diagram showing a fourth measurement result according to Example 4 of the present invention. [Figure 14] FIG. 11 is a diagram showing a first measurement result according to Example 5 of the present invention. [Figure 15] FIG. 11 is a diagram showing a second measurement result according to Example 5 of the present invention. [Figure 16] FIG. 13 is a diagram showing the measurement results according to Example 6 of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] Hereinafter, an embodiment of the present invention will be described with reference to the drawings. However, the present invention is not limited to the following embodiment. Note that, in the drawings, the same or equivalent parts are given the same reference symbols and the description will not be repeated.

[0020] [Embodiment 1] First, referring to Fig. 1, a particle analysis method and a particle manufacturing method according to the present embodiment will be described. Fig. 1 is a flow chart showing the particle manufacturing method according to the present embodiment. As shown in Fig. 1, the particle manufacturing method according to the present embodiment includes a first particle preparation step S1, a first observation step S2, a first migration speed measurement step S3, a first magnetic susceptibility measurement step S4, an encapsulation process step S5, a second observation step S6, a second migration speed measurement step S7, a second magnetic susceptibility measurement step S8, and a magnetic susceptibility analysis step S9. Among these steps S1 to S9, the first observation step S2, the first migration speed measurement step S3, the first magnetic susceptibility measurement step S4, the encapsulation process step S5, the second observation step S6, the second migration speed measurement step S7, the second magnetic susceptibility measurement step S8, and the magnetic susceptibility analysis step S9 represent the particle analysis method according to the present embodiment.

[0021] <First particle preparation step S1> In the first particle preparation step S1, the first particle p1 is prepared. The first particle p1 has a particle diameter of, for example, 1 nm or more and several hundred μm or less. The first particle p1 is, for example, a base material of a microcapsule (capsule base material). The capsule base material has a membrane component that separates the outside from the inside. However, the first particle p1 is not limited to a capsule base material. The first particle p1 may be any particle that can encapsulate the component pt to be encapsulated.

[0022] The first particle p1 may be, for example, a capsule substrate derived from yeast. For example, the capsule substrate derived from yeast is made of solid components of yeast remaining after the intracellular components are released from the yeast. In this case, the capsule substrate derived from yeast is mainly made of the skeletal portion of yeast. More specifically, the capsule substrate derived from yeast has a cell wall and cell membrane of yeast. The intracellular components mainly include amino acids, nucleic acid-related substances, minerals, and vitamins.

[0023] The yeast-derived capsule material is produced from, for example, torula yeast, baker's yeast, brewer's yeast, or sake yeast. The form of the yeast-derived capsule material is not particularly limited, and may be, for example, any of compressed yeast, dry yeast, active dry yeast, dead yeast, and sterilized dry yeast.

[0024] The capsule base material derived from yeast may be a substance derived from the solid component of yeast. The substance derived from the solid component of yeast has substantially the same composition as the solid component of yeast. Specifically, the capsule base material derived from yeast may be a pulverized or powdered product of the solid component of yeast.

[0025] The yeast used in the production of the yeast-derived capsule material is not particularly limited, and may be, for example, a yeast belonging to the genus Saccharomyces or a yeast belonging to the genus Candida. For example, the yeast may be Saccharomyces cerevisiae. Alternatively, the yeast may be Candida utilis.

[0026] The first particle p1 is not limited to a capsule base material derived from yeast. The first particle p1 may be, for example, yeast, algae such as Euglena, lactic acid bacteria, acetic bacteria, Bacillus subtilis, or cell membrane fragments. The yeast, Euglena, lactic acid bacteria, acetic bacteria, and Bacillus subtilis may be alive or dead. For example, the yeast, algae such as Euglena, lactic acid bacteria, acetic bacteria, and Bacillus subtilis may be dried before or after sterilization. The first particle p1 is not limited to a microorganism, and may be an animal cell or a plant cell.

[0027] Alternatively, the first particle p1 may be a vesicle. More specifically, the first particle p1 may be, for example, a liposome. The surface membrane of the vesicle is made of lipids. More specifically, the surface membrane of the vesicle has a lipid bilayer. The liposome is a vesicle whose surface membrane is made of phospholipids. The liposome is used as an encapsulating material in a DDS (drug delivery system). The encapsulating material used in the DDS encapsulates, for example, a physiologically active substance as the component pt to be encapsulated.

[0028] <First particle manufacturing process> The first particle preparation step S1 may include a first particle production step of producing the first particle p1. When the first particle p1 is a capsule base material derived from yeast, the first particle production step includes a step of releasing intracellular components from the yeast. The treatment for releasing intracellular components from the yeast is not particularly limited, and may be, for example, any of a hot water treatment method, an autolysis method, and an enzymatic decomposition method.

[0029] For example, the hot water treatment method is a process in which a high-temperature solution is stirred for a predetermined period of time. Yeast is added to the solution as a solute. The solvent is, for example, distilled water. However, the solvent is not limited to distilled water. The solvent may be any liquid that can release intracellular components from the yeast. For example, a buffer solution or an emulsifier may be used as the solvent.

[0030] When the intracellular components are released from the yeast by the hot water treatment method, the solid components remaining after the intracellular components are released from the yeast may be subjected to an enzyme treatment by adding a protease and / or cellulase. Furthermore, an emulsifier may be added to the solid components. The timing of adding the emulsifier may be before or after the enzyme treatment. Alternatively, the timing of adding the emulsifier may be the same as the enzyme treatment.

[0031] The solid components remaining after the intracellular components are released from the yeast may be sterilized. That is, the capsule base material derived from the yeast may be sterilized. In addition, the solid components may be dried in the final stage of the first particle manufacturing process. That is, the capsule base material derived from the yeast may be in a dry state. However, the first particles p1 are not limited to being in a dry state. The first particles p1 may be in a paste state containing an appropriate amount of moisture.

[0032] <First observation step S2> In the first observation step S2, the first particles p1 are magnetophoretically migrated and observed. Specifically, the magnetophoretically migrated first particles p1 are observed using an analysis device 10 described with reference to FIG.

[0033] <First migration speed measurement step S3> In the first migration velocity measurement step S3, the magnetophoretic velocity v1 of the first particle p1 is measured from the results of the observation in the first observation step S2. Specifically, the magnetophoretic velocity v1 of the first particle p1 is measured using an analysis device 10 described with reference to Fig. 2. In this embodiment, the particle diameter d1 of the first particle p1 is measured from the results of the observation in the first observation step S2 together with the magnetophoretic velocity v1 of the first particle p1.

[0034] <First magnetic susceptibility measurement step S4> In the first magnetic susceptibility measurement step S4, the volume magnetic susceptibility χs1 of the first particle p1 is measured based on the result of the measurement in the first migration velocity measurement step S3. Specifically, the volume magnetic susceptibility χs1 of the first particle p1 is measured using an analysis device 10 described with reference to FIG.

[0035] <Encapsulation processing step S5> In the encapsulation process S5, an encapsulation process is performed to encapsulate a component pt to be encapsulated, which is smaller than the first particle p1, in the first particle p1 to obtain a second particle p2. The second particle p2 is the first particle p1 after the encapsulation process. More specifically, in the encapsulation process S5, the first particle p1 and the component pt to be encapsulated are mixed to contact the first particle p1 and the component pt to be encapsulated. Here, the first particle p1 to be encapsulated may be a first particle p1 whose volume magnetic susceptibility χs1 has been measured, or a first particle p1 whose volume magnetic susceptibility χs1 has not been measured. The second particle p2 may be, for example, a microcapsule. For example, when the first particle p1 is a capsule base material derived from yeast, the second particle p2 is a microcapsule.

[0036] The component pt to be encapsulated is not particularly limited as long as it can be encapsulated in the first particle p1. The component pt to be encapsulated may be a liquid or a solid. The component pt to be encapsulated may be, for example, a pharmaceutical, an agricultural chemical, a flavoring, or a food ingredient.

[0037] Specifically, the encapsulated component pt can be a physiologically active substance, a mineral, a flavoring, a spice extract, a flavor oil, an animal fat, or a vegetable fat. For example, flavorings include d-limonene, carvone, ethyl caproate, chili flavor, vanilla flavor, grill flavor, wasabi flavor, coffee flavor, pepper, black pepper, mustard, curry spice, and meat flavor.

[0038] Specifically, the component pt to be encapsulated may be a hydrophobic substance or a hydrophilic substance. The component pt to be encapsulated may be a fat-soluble substance or a non-fat-soluble substance. When the first particle p1 has a hydrophobic region therein, the hydrophobic substance is encapsulated in the first particle p1 more stably than the hydrophilic substance. Similarly, when the first particle p1 has a hydrophilic region therein, the hydrophilic substance is encapsulated in the first particle p1 more stably than the hydrophobic substance.

[0039] The hydrophobic substance may be, for example, at least one derived lipid selected from the group consisting of long chain hydrocarbons, long chain alcohols, long chain amino alcohols, long chain aldehydes, long chain ketones, long chain acids and their salts, terpenoids, steroids, and carotenoids. Alternatively, the hydrophobic substance may be, for example, at least one simple lipid selected from the group consisting of waxes, glycerides, ether glycerides, and ceramides. Alternatively, the hydrophobic substance may be, for example, at least one complex lipid selected from the group consisting of phospholipids, glycolipids, phosphoglycolipids, sulfolipids, and amino acid lipids. The hydrophilic substance may be, for example, at least one substance selected from the group consisting of amino acids, water-soluble vitamins, nucleotides, and sugars.

[0040] The encapsulation process is not particularly limited as long as it is a process for encapsulating the component pt to be encapsulated in the first particle p1. For example, the encapsulation process may be a process for mixing and stirring the first particle p1 and the component pt to be encapsulated in a solvent. The solvent is, for example, distilled water. However, the solvent is not limited to distilled water. The solvent may be any liquid that can mix and disperse the first particle p1 and the component pt to be encapsulated. For example, the solvent may be a buffer solution, sugar solution, salt solution, seasoning solution, or soup stock. When the solvent contains salt, the amount of the component pt to be encapsulated in the first particle p1 can be increased compared to when the solvent does not contain salt.

[0041] The encapsulation process may include a process of drying a dispersion liquid obtained by mixing the first particles p1 and the component pt to be encapsulated in a solvent. By drying the dispersion liquid, the second particles p2 in a dry state are obtained. The process of drying the dispersion liquid is not particularly limited as long as the second particles p2 in a dry state can be obtained. For example, the dispersion liquid can be dried by spray drying, heat drying, freeze drying, or vacuum drying.

[0042] The encapsulation process is not limited to a process of mixing and stirring the first particle p1 and the component pt to be encapsulated in a solvent. For example, the encapsulation process may be a process of adjusting an environmental factor of a system in which the first particle p1 and the component pt to be encapsulated are dispersed. The environmental factors include heat, pressure, ion concentration, pH value, and the like. For example, by adjusting the ion concentration, the component pt to be encapsulated can be moved into the first particle p1 by the effect of coexisting ions. Specifically, the component pt to be encapsulated moves into the first particle p1 according to the second law of thermodynamics. When the first particle p1 is a living yeast and the component pt to be encapsulated is zinc (a mineral), the encapsulation process may be, for example, a process of culturing the yeast in a medium or culture solution to which zinc has been added.

[0043] <Second observation step S6> In the second observation step S6, the second particles p2 to be analyzed, which are the first particles p1 after the encapsulation process, are magnetophoresed and observed. Specifically, similarly to the first observation step S2, the magnetophoresed second particles p2 are observed using the analysis device 10 described with reference to FIG.

[0044] <Second migration speed measurement step S7> In the second migration velocity measurement step S7, the magnetophoretic velocity v2 of the second particle p2 is measured from the results of the observation in the second observation step S6. Specifically, similar to the first migration velocity measurement step S3, the magnetophoretic velocity v2 of the second particle p2 is measured using an analysis device 10 described with reference to Fig. 2. In this embodiment, the particle diameter d2 of the second particle p2 is measured from the results of the observation in the second observation step S6 together with the magnetophoretic velocity v2 of the second particle p2.

[0045] <Second magnetic susceptibility measurement step S8> In the second magnetic susceptibility measurement step S8, the volume magnetic susceptibility χs2 of the second particle p2 is measured based on the measurement result in the second migration velocity measurement step S7. Specifically, similar to the first magnetic susceptibility measurement step S4, the volume magnetic susceptibility χs2 of the second particle p2 is measured using the analysis device 10 described with reference to FIG.

[0046] <Magnetic susceptibility analysis step S9> In the magnetic susceptibility analysis step S9, it is analyzed whether or not the component pt to be included is included in the second particle p2 based on the result of the measurement in the first magnetic susceptibility measurement step S4 and the result of the measurement in the second magnetic susceptibility measurement step S8. Specifically, the difference between the volume magnetic susceptibility χs1 of the first particle p1 and the volume magnetic susceptibility χs2 of the second particle p2 is analyzed. When the volume magnetic susceptibility χs1 of the first particle p1 and the volume magnetic susceptibility χs2 of the second particle p2 show different values, it is determined that the component pt to be included is included in the second particle p2.

[0047] Specifically, the volume magnetic susceptibility χs2 of the second particle p2 differs depending on the component of the second particle p2. Therefore, the volume magnetic susceptibility χs2 of the second particle p2 differs depending on whether the second particle p2 contains the component pt to be contained. In other words, the volume magnetic susceptibility χs2 of the second particle p2 containing the component pt to be contained differs from the volume magnetic susceptibility χs2 of the second particle p2 not containing the component pt to be contained. Specifically, by containing the component pt to be contained, the volume magnetic susceptibility χs2 of the second particle p2 becomes closer to the volume magnetic susceptibility χst of the component pt to be contained compared to the volume magnetic susceptibility χs2 of the second particle p2 not containing the component pt to be contained.

[0048] Therefore, by comparing the volume magnetic susceptibility χs1 of the first particle p1 with the volume magnetic susceptibility χs2 of the second particle p2, it can be determined whether or not the component pt to be included is included in the second particle p2. More specifically, if the volume magnetic susceptibility χs2 of the second particle p2 is closer to the volume magnetic susceptibility χst of the component pt to be included than the volume magnetic susceptibility χs1 of the first particle p1, it is determined that the component pt to be included is included in the second particle p2.

[0049] The second particles p2 may be dried after the measurement of the volume magnetic susceptibility χs2. For example, after the second observation step S6, the solution (dispersion) in which the second particles p2, which are a solute (dispersoid), are dispersed is dried to obtain the second particles p2 in a dry state. The process for drying the dispersion is not particularly limited as long as the second particles p2 in a dry state can be obtained. For example, the dispersion can be dried by spray drying, heat drying, freeze drying, or vacuum drying.

[0050] <Analyzer 10> Next, an analysis device 10 according to the present embodiment will be described with reference to Fig. 1 and Fig. 2. Fig. 2 is a schematic diagram of the analysis device 10 according to the present embodiment. The analysis device 10 analyzes whether or not a component pt to be included is included in the second particle p2.

[0051] As shown in Fig. 2, the analysis device 10 includes a magnetic field generating unit 20, an observation unit 30, and a calculation unit 40. A cell 21 is disposed near the magnetic field generating unit 20. The magnetic field generating unit 20 generates a magnetic field to magnetophoretically migrate particles p in the cell 21. The observation unit 30 observes the particles p in the cell 21. The calculation unit 40 measures the particle diameter d and magnetophoretic velocity v of the particles p from the results of observation by the observation unit 30. The calculation unit 40 also measures the volume magnetic susceptibility χs of the particles p based on the particle diameter d and magnetophoretic velocity v of the particles p.

[0052] Specifically, in a first observation step S2, the magnetic field generation unit 20 magnetophores the first particle p1 in the cell 21, and the observation unit 30 observes the first particle p1 in the cell 21. In a first migration velocity measurement step S3, the calculation unit 40 measures the magnetophoretic velocity v1 of the first particle p1 from the result of observation by the observation unit 30. In this embodiment, the calculation unit 40 measures the particle diameter d1 of the first particle p1 as well as the magnetophoretic velocity v1 of the first particle p1 from the result of observation by the observation unit 30. In addition, in a first magnetic susceptibility measurement step S4, the calculation unit 40 measures the volume magnetic susceptibility χs1 of the first particle p1 based on the magnetophoretic velocity v1 and particle diameter d1 of the first particle p1.

[0053] Furthermore, in a second observation step S6, the magnetic field generation unit 20 magnetophoretically migrates the second particle p2 in the cell 21, and the observation unit 30 observes the second particle p2 in the cell 21. In a second migration velocity measurement step S7, the calculation unit 40 measures the magnetophoretic velocity v2 of the second particle p2 from the result of observation by the observation unit 30. In this embodiment, the calculation unit 40 measures the particle diameter d2 of the second particle p2 as well as the magnetophoretic velocity v2 of the second particle p2 from the result of observation by the observation unit 30. In addition, in a second magnetic susceptibility measurement step S8, the calculation unit 40 measures the volume magnetic susceptibility χs2 of the second particle p2 based on the magnetophoretic velocity v2 and particle diameter d2 of the second particle p2.

[0054] Furthermore, in the magnetic susceptibility analysis step S9, the calculation unit 40 analyzes whether or not the component pt to be included is included in the second particle p2 based on the volume magnetic susceptibility χs1 of the first particle p1 and the volume magnetic susceptibility χs2 of the second particle p2.

[0055] The analysis device 10 will be described in more detail below. The magnetic field generating unit 20 generates a magnetic field gradient (magnetic flux density gradient) to apply a magnetic force to the particles p in the cell 21. As a result, the particles p undergo magnetophoresis. In this embodiment, the magnetic field generating unit 20 includes two permanent magnets 20a, 20b (see FIG. 3) that generate a magnetic field gradient. The two permanent magnets 20a, 20b are arranged with a gap of a certain distance, for example, 100 μm or more and 500 μm or less. The cell 21 is arranged in the gap between the two permanent magnets 20a, 20b.

[0056] In this embodiment, the cell 21 is a capillary tube. The capillary tube is an example of a tubular member. The material of the cell 21 is not particularly limited as long as it is a material that can transmit visible light or laser light. For example, the cell 21 may be made of glass or plastic.

[0057] The particles p are introduced into the cell 21 together with the medium m by, for example, a microsyringe, a micropump, or an autosampler. Alternatively, the particles p may be introduced into the cell 21 together with the medium m based on the siphon principle. Alternatively, a droplet containing the particles p may be introduced into the cell 21 (capillary tube) by capillary action. When the droplet containing the particles p is dropped onto one end of the capillary tube, the droplet flows through the capillary tube by capillary action.

[0058] A particle p exists in a medium m. A single particle p may exist in the medium m, or multiple particles p may exist in the medium m. When multiple particles p exist in the medium m, the multiple particles p may be dispersed in the medium m, or may be unevenly distributed in the medium m. The medium m is, for example, distilled water. The analysis device 10 measures the volume magnetic susceptibility χs of each of the multiple particles p existing in the medium m.

[0059] The observation unit 30 observes the particle p in the cell 21 and generates a signal indicating the observation result. The calculation unit 40 measures the magnetophoretic velocity v and particle diameter d of the particle p based on the signal generated by the observation unit 30. The calculation unit 40 includes a storage unit 41, a processing unit 42, and an output unit 43.

[0060] The storage unit 41 stores programs, setting information, and the like. The storage unit 41 may be configured with, for example, a storage device and a semiconductor memory. The storage device is, for example, a hard disk drive (HDD). The storage unit 41 may have, as the semiconductor memory, for example, a random access memory (RAM) and a read only memory (ROM).

[0061] The processing unit 42 executes the programs stored in the memory unit 41 to perform various processes such as numerical calculations, information processing, and device control. The processing unit 42 is configured with a processor such as a CPU (Central Processing Unit) or a microcomputer. Alternatively, the processing unit 42 is configured with an integrated circuit such as an ASIC (Application Specific Integrated Circuit). The integrated circuit has a logic circuit. The processing unit 42 may be configured with a processor and an integrated circuit. Note that a general-purpose computer such as a personal computer may be used as the calculation unit 40.

[0062] The processing unit 42 measures the change over time in the position of the particle p in the cell 21 from the observation result of the observation unit 30 (the signal generated by the observation unit 30). For example, the processing unit 42 measures the position of the particle p in the cell 21 at each predetermined time interval. In other words, the processing unit 42 measures the position of the particle p at different times. The processing unit 42 measures the magnetophoretic velocity v of the particle p from the change over time in the position of the particle p.

[0063] In addition, the processing unit 42 measures the particle diameter d of the particle p from the signal generated by the observation unit 30. The processing unit 42 further measures the volume magnetic susceptibility χs of the particle p based on the magnetophoretic velocity v and the particle diameter d of the particle p.

[0064] For example, the processing unit 42 calculates the volume magnetic susceptibility χs of the particle p based on the following formula (1). v={2(χs-χm)r 2 / 9ημ o}B(dB / dx)···(1)

[0065] In equation (1), v is the magnetophoretic velocity of particle p, χs is the volume magnetic susceptibility of particle p, χm is the volume magnetic susceptibility of medium m, r is the radius of particle p, η is the viscosity of medium m, and μ o is the magnetic permeability of a vacuum, B is the magnetic flux density, and dB / dx is the magnetic field gradient (gradient of the magnetic flux density). Equation (1) is derived from the fact that the difference in the magnetic force acting on the particle p and the medium m in the axial direction (x direction) of the cell 21 (capillary tube) is approximately equal to the viscous drag force.

[0066] The magnetophoretic velocity v and the volume magnetic susceptibility χs are values ​​measured by the processing unit 42. The radius r of the particle p is calculated, for example, by the processing unit 42 from the measured particle diameter d of the particle p. The volume magnetic susceptibility χm of the medium m, the viscosity η of the medium m, and the magnetic permeability μ of a vacuum o , magnetic flux density B, and magnetic field gradient dB / dx are stored in advance in the storage unit 41. For example, the analyst operates an input device such as a keyboard, a mouse, or a touch display to input the volume magnetic susceptibility χm of the medium m, the viscosity η of the medium m, the magnetic permeability μ of a vacuum, o , the magnetic flux density B, and the magnetic field gradient dB / dx can be stored in the storage unit 41. The volume magnetic susceptibility χm of the medium m, the viscosity η of the medium m, the magnetic permeability μ o are, for example, literature values. The magnetic flux density B and the magnetic field gradient dB / dx are, for example, measured values.

[0067] The storage unit 41 stores the volume magnetic susceptibility χs1 of the first particle p1 measured in the first magnetic susceptibility measurement step S4. The storage unit 41 also stores the volume magnetic susceptibility χs2 of the second particle p2 measured in the second magnetic susceptibility measurement step S8. The processing unit 42 accesses the storage unit 41 and acquires the volume magnetic susceptibility χs1 of the first particle p1 and the volume magnetic susceptibility χs2 of the second particle p2 from the storage unit 41. The processing unit 42 then compares the volume magnetic susceptibility χs1 of the first particle p1 with the volume magnetic susceptibility χs2 of the second particle p2 to analyze whether or not the component pt to be included is included in the second particle p2. More specifically, if the volume magnetic susceptibility χs2 of the second particle p2 is closer to the volume magnetic susceptibility χst of the component pt to be encapsulated than the volume magnetic susceptibility χs1 of the first particle p1, the processing unit 42 determines that the component pt to be encapsulated is contained in the second particle p2.

[0068] The volume magnetic susceptibility χst of the component pt of the inclusion target is stored in advance in the storage unit 41. For example, the volume magnetic susceptibility χst of the component pt of the inclusion target may be a literature value. In this case, the analyst operates the input device to store the volume magnetic susceptibility χst (literature value) of the component pt of the inclusion target in the storage unit 41. Alternatively, the analyst may use the analysis device 10 to measure the volume magnetic susceptibility χst of the component pt of the inclusion target. In this case, the volume magnetic susceptibility χst of the component pt of the inclusion target measured by the processing unit 42 is stored in the storage unit 41.

[0069] The output unit 43 outputs the analysis result (determination result) by the processing unit 42. Specifically, the output unit 43 outputs information indicating whether or not the component pt to be encapsulated is encapsulated in the second particle p2. The output unit 43 includes, for example, a display. In this case, the processing unit 42 displays information indicating the analysis result on the display. The output unit 43 may include a communication interface in addition to the display or instead of the display. The communication interface is, for example, a USB (Universal Serial Bus) connector. In this case, the processing unit 42 outputs a signal indicating the analysis result to an external device via the communication interface and a USB cable. The external device is, for example, a display or a printer. When the external device is a display, information indicating the analysis result is displayed on the external device. When the external device is a printer, an image indicating the analysis result is printed. Alternatively, the external device may be a USB memory. In this case, the processing unit 42 outputs a signal indicating the analysis result to the USB memory via the communication interface. As a result, the information indicating the analysis result is stored in the USB memory.

[0070] Next, the movement of the particle p will be described with reference to Fig. 3(a) and Fig. 3(b). Fig. 3(a) and Fig. 3(b) are diagrams showing the movement of the particle p. As shown in Fig. 3(a) and Fig. 3(b), the magnetic field generating unit 20 includes a permanent magnet 20a whose magnetic pole is a north pole and a permanent magnet 20b whose magnetic pole is a south pole. The two permanent magnets 20a, 20b face each other with the cell 21 between them.

[0071] As shown in Fig. 3(a), when the volume magnetic susceptibility χs of the particle p is smaller than the volume magnetic susceptibility χm of the medium m, the particle p moves in a direction away from the magnetic field (magnetic field generating unit 20). On the other hand, as shown in Fig. 3(b), when the volume magnetic susceptibility χs of the particle p is larger than the volume magnetic susceptibility χm of the medium m, the particle p moves in a direction toward the magnetic field (magnetic field generating unit 20).

[0072] 3(a) and 3(b), the movement of particle p is determined according to the relationship between the volume magnetic susceptibility χs of particle p and the volume magnetic susceptibility χm of medium m. Note that particle p receives force in the vicinity of the ends of permanent magnets 20a and 20b. For example, particle p receives force within a range of about ±200 μm from the vicinity of the ends of permanent magnets 20a and 20b.

[0073] Next, the analysis device 10 will be further described with reference to Fig. 4. Fig. 4 is a diagram showing the configuration of the analysis device 10. As shown in Fig. 4, the analysis device 10 further includes a light source 50. In addition, the observation unit 30 includes a magnifying unit 32 and an imaging unit 34.

[0074] The light source 50 emits light of relatively high intensity including a visible light component. The light source 50 irradiates the cell 21 with light. As a result, the particle p is irradiated with light. The wavelength spectrum of the light emitted from the light source 50 may be relatively broad. For example, a halogen lamp is suitably used as the light source 50.

[0075] The particle p introduced into the cell 21 is magnified at an appropriate magnification by the magnifying section 32 and imaged by the imaging section 34. The position of the particle p can be identified from the imaging result of the imaging section 34 (the image captured by the imaging section 34). For example, the magnifying section 32 includes an objective lens, and the imaging section 34 includes a charge coupled device (CCD). Alternatively, each pixel of the imaging section 34 may be composed of a photodiode or a photomultiplier tube. The imaging section 34 images the particle p at, for example, a predetermined time interval. Note that the imaging section 34 may image light emitted from the light source 50 and transmitted through the cell 21, or may image light emitted from the light source 50 and scattered by the particle p.

[0076] The calculation unit 40 (processing unit 42) measures the change in position of the particle p over time from the imaging result of the imaging unit 34, and measures the magnetophoretic velocity v of the particle p from the change in position of the particle p over time.

[0077] The calculation unit 40 (processing unit 42) also measures the particle diameter d of the particle p from the imaging result of the particle p. For example, the calculation unit 40 (processing unit 42) executes the following process. That is, first, the image captured by the imaging unit 34 is converted to monochrome and its brightness is quantified. Next, the differential value of the brightness value is compared with a threshold value to set the boundary of the particle p. Next, the area of ​​the particle p is detected from the set boundary, and the particle diameter d is calculated from the radius of a circle corresponding to the area. Alternatively, the center of the particle p is specified, multiple straight lines are drawn that pass through the center of the particle p, and the average distance between two points where each straight line intersects with the boundary of the particle p is calculated.

[0078] The first embodiment of the present invention has been described above with reference to Fig. 1 to Fig. 4. According to this embodiment, it is possible to analyze for each analysis target whether or not the particle to be analyzed (second particle p2) contains the component pt to be contained therein.

[0079] The particles p for which the volume magnetic susceptibility χs1 is measured may be all or a part of the first particles p1 prepared in the first particle preparation step S1. Similarly, the particles p for which the volume magnetic susceptibility χs2 is measured may be all or a part of the second particles p2 obtained in the encapsulation step S5.

[0080] [Embodiment 2] Next, a second embodiment of the present invention will be described with reference to Fig. 5. However, differences from the first embodiment will be described, and a description of the same matters as in the first embodiment will be omitted. The second embodiment differs from the first embodiment in that the second particle p2 is spectroscopically analyzed.

[0081] 5 is a flow chart showing the particle production method according to the present embodiment. The particle production method according to the present embodiment further includes a spectroscopic analysis step S10.

[0082] <Spectroscopic analysis process S10> In the spectroscopic analysis step S10, the second particle p2 is observed spectroscopically to analyze whether or not the component pt to be included is included in the second particle p2. Specifically, the second particle p2 is irradiated with laser light, the scattered light scattered from the second particle p2 is detected, and the spectrum of the scattered light is analyzed. By analyzing the spectrum of the scattered light scattered from the sample, the molecular structure of the sample can be identified, and therefore, it is possible to analyze whether or not the component pt to be included is included in the second particle p2 from the scattered light scattered from the second particle p2. Specifically, it is analyzed whether or not the spectrum of the scattered light includes a spectrum indicating the component pt to be included.

[0083] For example, by analyzing the Raman spectrum using a Raman spectrometer, it is possible to analyze whether or not the component pt to be contained in the second particle p2 is contained. The Raman spectrometer may include a microscope. In other words, the Raman spectrometer may be a confocal Raman microscope.

[0084] Specifically, the Raman spectroscopy device includes a laser light source, a detector, and a calculation unit. The laser light source irradiates the second particle p2 with laser light to generate scattered light from the second particle p2. The detector detects the scattered light. The calculation unit of the Raman spectroscopy device analyzes the spectrum of the scattered light based on the output of the detector. In detail, the calculation unit identifies the molecular structure of the second particle p2 by analyzing the spectrum of the scattered light. The calculation unit includes, for example, a microcomputer or an ASIC.

[0085] The scattered light spectrum may be analyzed using the calculation unit 40 described with reference to Fig. 2. More specifically, the output from the detector of the Raman spectrometer is input to the calculation unit 40. A processing unit 42 of the calculation unit 40 analyzes the scattered light spectrum based on the detector output, and analyzes whether or not the scattered light spectrum contains a spectrum indicating the component pt to be included, based on the analysis result.

[0086] The second embodiment of the present invention has been described above with reference to Fig. 5. According to this embodiment, in addition to the analysis based on the volume magnetic susceptibility χs1 of the first particle p1 and the volume magnetic susceptibility χs2 of the second particle p2, it is possible to spectroscopically analyze whether or not the component pt to be included is included in the second particle p2. Therefore, it is possible to more reliably analyze for each analysis target whether or not the component pt to be included is included in the particle to be analyzed (second particle p2).

[0087] The particles p to be analyzed spectroscopically may be all or a part of the second particles p2 obtained in the encapsulation process S5.

[0088] 5, the spectroscopic analysis step S10 is performed after the magnetic susceptibility analysis step S9, but the order of performing the spectroscopic analysis step S10 is not limited to after the magnetic susceptibility analysis step S9. The spectroscopic analysis step S10 may be performed, for example, before the second observation step S6.

[0089] In addition, in this embodiment, a Raman spectrometer is used, but the spectrometer is not limited to a Raman spectrometer. For example, an ultraviolet-visible spectrometer or an infrared spectrometer may be used. When an ultraviolet-visible spectrometer is used, an ultraviolet-visible absorption spectrum is analyzed to analyze whether or not the component pt to be included is included in the second particle p2. When an infrared spectrometer is used, an infrared absorption spectrum is analyzed to analyze whether or not the component pt to be included is included in the second particle p2. Alternatively, a near-infrared spectrum may be obtained using an infrared spectrometer, and the near-infrared spectrum may be analyzed by statistical processing to analyze whether or not the component pt to be included is included in the second particle p2. In other words, a near-infrared spectrum may be analyzed chemometrically to analyze whether or not the component pt to be included is included in the second particle p2.

[0090] [Embodiment 3] Next, a third embodiment of the present invention will be described with reference to Fig. 6. However, differences from the first and second embodiments will be described, and a description of the same aspects as the first and second embodiments will be omitted. The third embodiment differs from the first and second embodiments in that the zeta potentials of the first particle p1 and the second particle p2 are analyzed.

[0091] 6 is a flowchart showing the particle production method according to this embodiment. The particle production method according to this embodiment further includes a first zeta potential measurement step S11, a second zeta potential measurement step S12, and a zeta potential analysis step S13.

[0092] <First zeta potential measurement step S11> In the first zeta potential measurement step S11, the zeta potential of the first particle p1 is measured. More specifically, the first particle p1 is electrophoresed in a medium m, and the electrophoretic velocity of the first particle p1 is measured. Then, the zeta potential of the first particle p1 is measured based on the electrophoretic velocity.

[0093] Specifically, the zeta potential is measured using a zeta potential measuring device. The zeta potential measuring device includes a positive electrode and a negative electrode. A cell is placed between the positive electrode and the negative electrode. A solution in which first particles p1, which are solutes (dispersoids), are dispersed is enclosed in the cell. The zeta potential measuring device applies a voltage to the positive electrode to generate an electric field between the positive electrode and the negative electrode. As a result, the first particles p1 undergo electrophoresis. Zeta potential measuring devices include devices that measure the average value of the zeta potential of all particles p dispersed in the cell, and devices that measure the zeta potential of each particle p dispersed in the cell.

[0094] The device for measuring the average value of the zeta potential further includes a laser light source, a detector, and a calculation unit. The laser light source irradiates the entire first particle p1 undergoing electrophoresis with laser light to generate scattered light from the entire first particle p1. The detector detects the scattered light. More specifically, the frequency of the scattered light generated from the entire first particle p1 undergoing electrophoresis shifts (changes) due to the Doppler effect. The detector detects the frequency of the scattered light that shifts (changes) due to the Doppler effect.

[0095] The calculation unit of the device for measuring the average value of the zeta potential measures the average value of the electrophoretic velocity based on the output of the detector. Specifically, the amount of shift (change) in the frequency of the scattered light is proportional to the average value of the electrophoretic velocity. The calculation unit measures the average value of the electrophoretic velocity of the entire first particle p1 by measuring the amount of shift in the frequency of the scattered light. Then, the calculation unit measures the average value of the zeta potential based on the average value of the electrophoretic velocity. The calculation unit includes, for example, a microcomputer or an ASIC. Note that the calculation unit 40 described with reference to FIG. 2 may be used as the calculation unit.

[0096] The device for measuring the individual zeta potentials of the particles p further includes a laser light source, a detector, and a calculation unit, similar to the device for measuring the average value of the zeta potential. The laser light source irradiates the entire first particle p1 undergoing electrophoresis with laser light to generate scattered light from the entire first particle p1. The detector includes an objective lens and an image sensor. The image sensor may be a semiconductor sensor such as a CCD image sensor or a CMOS image sensor. The detector captures the light magnified by the objective lens with the image sensor. As a result, the movement of each of the first particles p1 is captured.

[0097] A calculation unit of the device for measuring the individual zeta potentials of the particles p measures the electrophoretic velocity of each of the first particles p1 based on an image signal output from a detector. The calculation unit then measures the zeta potential of each of the first particles p1 based on the electrophoretic velocity. The calculation unit includes, for example, a microcomputer or an ASIC. Note that the calculation unit 40 described with reference to FIG. 2 may be used as the calculation unit.

[0098] <Second zeta potential measurement step S12> In the second zeta potential measurement step S12, the zeta potential of the second particle p2 is measured. The second zeta potential measurement step S12 is similar to the first zeta potential measurement step S11, except that the measurement target is the second particle p2.

[0099] <Zeta potential analysis step S13> In the zeta potential analysis step S13, it is analyzed whether or not the component pt to be included is included in the second particle p2 based on the result of the measurement in the first zeta potential measurement step S11 and the result of the measurement in the second zeta potential measurement step S12. Specifically, the zeta potential of the first particle p1 is compared with the zeta potential of the second particle p2 to analyze whether or not there is a difference between the zeta potential of the first particle p1 and the zeta potential of the second particle p2. When the zeta potential of the first particle p1 and the zeta potential of the second particle p2 show the same value, it is determined that the component pt to be included is included in the second particle p2.

[0100] Specifically, when the component pt to be encapsulated does not enter the second particle p2 but adheres to the surface of the second particle p2, the zeta potential of the second particle p2 exhibits a value different from that of the first particle p1. In other words, when the component pt to be encapsulated does not adhere to the surface of the second particle p2 but enters the second particle p2, the zeta potential of the second particle p2 exhibits the same value as that of the first particle p1. Therefore, by comparing the zeta potential of the first particle p1 with the zeta potential of the second particle p2, it is possible to analyze whether the component pt to be encapsulated is encapsulated in the second particle p2.

[0101] The calculation unit 40 described with reference to FIG. 2 may be used to analyze the zeta potentials of the first particle p1 and the second particle p2. In detail, the analyst operates the input device to store data indicating the zeta potential of the first particle p1 and data indicating the zeta potential of the second particle p2 in the storage unit 41. Alternatively, the zeta potential measuring device and the calculation unit 40 are connected via a transmission cable such as a USB cable, and the data indicating the zeta potential of the first particle p1 and data indicating the zeta potential of the second particle p2 are transmitted from the zeta potential measuring device to the calculation unit 40. The processing unit 42 of the calculation unit 40 compares the zeta potential of the first particle p1 with the zeta potential of the second particle p2 to analyze whether the component pt to be included is included in the second particle p2.

[0102] The third embodiment of the present invention has been described above with reference to Fig. 6. According to this embodiment, in addition to the analysis based on the volume magnetic susceptibility χs1 of the first particle p1 and the volume magnetic susceptibility χs2 of the second particle p2 and the spectroscopic analysis, it is possible to analyze whether or not the component pt to be included is included in the second particle p2 from the zeta potentials of the first particle p1 and the second particle p2. Therefore, it is possible to more reliably analyze for each analysis target whether or not the component pt to be included is included in the particle to be analyzed (second particle p2).

[0103] The particles p whose zeta potential is to be measured in the first zeta potential measurement step S11 may be all or a part of the first particles p1 prepared in the first particle preparation step S1. Similarly, the particles p whose zeta potential is to be measured in the second zeta potential measurement step S12 may be all or a part of the second particles p2 obtained in the encapsulation treatment step S5.

[0104] 6, the first zeta potential measurement step S11 is performed after the first magnetic susceptibility measurement step S4, but the order of performing the first zeta potential measurement step S11 is not limited to the first magnetic susceptibility measurement step S4. The first zeta potential measurement step S11 may be performed, for example, before the first observation step S2.

[0105] 6, the second zeta potential measurement step S12 is performed after the second magnetic susceptibility measurement step S8, but the order of performing the second zeta potential measurement step S12 is not limited to the second magnetic susceptibility measurement step S8. The second zeta potential measurement step S12 may be performed, for example, before the second observation step S6.

[0106] 6, the zeta potential analysis step S13 is performed after the spectroscopic analysis step S10, but the order of performing the zeta potential analysis step S13 is not limited to after the spectroscopic analysis step S10. The zeta potential analysis step S13 may be performed after the second zeta potential measurement step S12. For example, the zeta potential analysis step S13 may be performed before the magnetic susceptibility analysis step S9.

[0107] Furthermore, in this embodiment, the spectroscopic analysis step S10 is performed, but the spectroscopic analysis step S10 may be omitted.

[0108] Although the embodiments of the present invention have been described above with reference to the drawings, the present invention is not limited to the above-described embodiments and can be embodied in various forms without departing from the spirit and scope of the present invention.

[0109] For example, in the embodiment according to the present invention, the particle diameter d of the particles p is measured by image analysis, but the particle diameter d of the particles p may be measured by analyzing the Brownian motion of the particles p.

[0110] Specifically, a diffusion coefficient is calculated from the variance of the change (displacement) in the position of particle p in the direction (y direction) perpendicular to the axial direction (x direction) of cell 21 (capillary tube), and the particle diameter d of particle p can be measured from this diffusion coefficient. In more detail, particle p is affected by the magnetic field gradient in the axial direction (x direction) of cell 21 (capillary tube), but is hardly affected by the magnetic field gradient in the direction (y direction) perpendicular to the axial direction of cell 21. Therefore, the diffusion coefficient D can be calculated from the variance of the displacement of the position of particle p in the y direction. Specifically, the diffusion coefficient D can be calculated by dividing the square of the movement distance in the y direction of particle p performing Brownian motion by twice the time.

[0111] The processing unit 42 measures the particle diameter d of the particle p from the diffusion coefficient D based on the following formula (2): In formula (2), d is the particle diameter of the particle p, k is the Boltzmann constant, T is the absolute temperature, and η is the viscosity of the medium m. d = kT / (3πηD) (2)

[0112] In the embodiment of the present invention, the analysis device 10 includes the light source 50. However, the analysis device 10 may include a laser light source instead of the light source 50, or may further include a laser light source in addition to the light source 50. When the analysis device 10 includes the light source 50 and the laser light source, when the light source 50 emits light, the laser light source stops emitting the laser light, and when the laser light source emits the laser light, the light source 50 stops emitting the light. When the laser light source is used, the particle p introduced into the cell 21 is irradiated with the laser light. The observation unit 30 observes the particle p by the laser light (scattered light) scattered by the particle p in the cell 21. For example, the imaging unit 34 described with reference to FIG. 3 captures an image of the laser light scattered by the particle p via the magnification unit 32.

[0113] When a laser light source is used, the particle diameter d of the particle p may be measured, for example, based on a dynamic light scattering method or a static light scattering method. When the particle p is irradiated with a laser beam, the capillary tube is preferably a square capillary having a square cross section perpendicular to the axial direction. By using a square capillary, it becomes easy to give a mirror finish to the side surface of the cell 21 that is irradiated with the laser beam.

[0114] In addition, in the embodiment of the present invention, the calculation unit 40 (processing unit 42) measures the particle diameter d of the particle p, but the image captured by the imaging unit 34 may be displayed on a display, and the analyst may measure the particle diameter d of the particle p from the image displayed on the display. Alternatively, the image captured by the imaging unit 34 may be printed, and the analyst may measure the particle diameter d of the particle p from the printed image. In this case, the analyst operates the input device to store data indicating the particle diameter d in the memory unit 41.

[0115] In the embodiment of the present invention, the particle diameter d is measured, but a literature value may be used as the particle diameter d. In this case, the analyst operates the input device to store data indicating the particle diameter in the memory unit 41.

[0116] In addition, in the embodiment of the present invention, the imaging unit 34 measures the magnetophoretic velocity v of the particle p by capturing an image of the particle p at a predetermined time interval. However, the magnetophoretic velocity v of the particle p may also be measured using a laser light source, for example, based on a laser Doppler method. EXAMPLES

[0117] The present invention will be described in more detail below using examples, but the scope of the present invention is not limited to the examples.

[0118] [Example 1] <Production of yeast-derived capsule materials> First, a capsule substrate D11 derived from yeast was produced. Hereinafter, the capsule substrate derived from yeast may be referred to as a "yeast capsule substrate". In Example 1, soluble components were removed from baker's yeast by hot water treatment to obtain a paste-like solid component, and the solid component was sterilized and dried to obtain the yeast capsule substrate D11. The yeast capsule substrate D11 was in a dry state. Then, the yeast capsule substrate D11 was made into a paste. Specifically, the dried yeast capsule substrate D11 was added to distilled water to prepare a solution L1, and the solution L1 was heated and sterilized. Then, the solution L1 was cooled to room temperature to make the yeast capsule substrate D11 into a paste.

[0119] <Cleaning process> The washing process was performed twice. Specifically, the yeast capsule substrate D11 in the form of a paste was added to distilled water and stirred to prepare a solution L2. Then, the distilled water was centrifuged from the solution L2 using a centrifuge to obtain the yeast capsule substrate D11 after the first washing process. Then, the yeast capsule substrate D11 after the first washing process was added to distilled water again and stirred to prepare a solution L3. Then, the distilled water was centrifuged from the solution L3 using a centrifuge to obtain the yeast capsule substrate D11 after the second washing process. Hereinafter, the "yeast capsule substrate D11 after the second washing process" and the like may be described as the "yeast capsule substrate D11 after washing" and the like. The yeast capsule substrate D11 after washing was in the form of a paste.

[0120] <Volume magnetic susceptibility measurement of yeast-derived capsule base material> The individual volume magnetic susceptibility of the first sample SP1 (yeast capsule substrate D11 after washing) was measured. Specifically, the first sample SP1 (yeast capsule substrate D11 after washing) was added to distilled water, and the distilled water was irradiated with ultrasonic waves (ultrasonic treatment) to prepare a solution L4. After ultrasonic treatment, the solution L4 was sealed in a cell, and the individual volume magnetic susceptibility of the first sample SP1 (yeast capsule substrate D11 after washing) was measured. In addition, after measuring the volume magnetic susceptibility, the solution L4 was sealed in a hemocytometer, and the number of the first sample SP1 (yeast capsule substrate D11 after washing) was counted using a microscope.

[0121] <Inclusion processing> The encapsulation process was carried out using the yeast capsule substrate D11 in the form of a paste, d-limonene, and distilled water. The mixing ratio of the yeast capsule substrate D11 in the form of a paste, d-limonene, and distilled water was 2:1:7 by weight. Here, the mixing ratio of d-limonene indicates the weight of d-limonene when the weight of the yeast capsule substrate D11 is set to "2". Similarly, the mixing ratio of distilled water indicates the weight of distilled water when the weight of the yeast capsule substrate D11 is set to "2".

[0122] Specifically, d-limonene was added to distilled water, and then the temperature was raised. Then, the yeast capsule substrate D11 in a paste form was added to the distilled water and stirred to prepare a solution L5. After stirring, the solution L5 was heated and sterilized. Then, the solution L5 was cooled to room temperature to prepare a paste-like microcapsule D12. The microcapsule D12 is the yeast capsule substrate D11 after the encapsulation treatment. Note that d-limonene is a hydrophobic substance, and the yeast capsule substrate D11 has a hydrophobic region inside.

[0123] <Cleaning process> The washing process was performed twice. Specifically, the paste-like microcapsules D12 were added to distilled water and stirred to prepare a solution L6. Then, the distilled water was centrifuged from the solution L6 using a centrifuge to obtain the microcapsules D12 after the first washing process. Then, the microcapsules D12 after the first washing process were added to distilled water again and stirred to prepare a solution L7. Then, the distilled water was centrifuged from the solution L7 using a centrifuge to prepare the microcapsules D12 after the second washing process. Hereinafter, the "microcapsules D12 after the second washing process" and the like may be described as the "microcapsules D12 after washing" and the like. The microcapsules D12 after washing were in a paste state.

[0124] <Volume magnetic susceptibility measurement of microcapsules> The volume magnetic susceptibility of each of the second sample SP2 (microcapsules D12 after washing) was measured. Specifically, the second sample SP2 was added to distilled water, and the distilled water was irradiated with ultrasonic waves (ultrasonic treatment) to prepare solution L8. After ultrasonic treatment, solution L8 was sealed in a cell, and the volume magnetic susceptibility of each of the second sample SP2 (microcapsules D12 after washing) was measured. In addition, after measuring the volume magnetic susceptibility, solution L8 was sealed in a hemocytometer, and the number of second sample SP2 (microcapsules D12 after washing) was counted using a microscope.

[0125] Table 1 shows the measurement results of Example 1. [Table 1]

[0126] FIG. 7 is a diagram showing the measurement results according to Example 1. In detail, FIG. 7 shows the measurement results of the volume magnetic susceptibility of the first sample SP1 (yeast capsule substrate D11 after washing) and the second sample SP2 (microcapsule D12 after washing). The first sample SP1 and the second sample SP2 correspond to the first particle p1 and the second particle p2 of the above embodiment, respectively. In FIG. 7, the horizontal axis shows the volume magnetic susceptibility, and the vertical axis shows the percentage (%) of the number of particles. In addition, in FIG. 7, the solid line graph shows the measurement results of the volume magnetic susceptibility of the first sample SP1 (yeast capsule substrate D11 after washing), and the dashed line graph shows the measurement results of the volume magnetic susceptibility of the second sample SP2 (microcapsule D12 after washing).

[0127] Specifically, the solid line graph was created by measuring the volume magnetic susceptibility of each of the first samples SP1 and calculating the ratio of the number of the first samples SP1 for each measured volume magnetic susceptibility. Similarly, the dashed line graph was created by measuring the volume magnetic susceptibility of each of the second samples SP2 and calculating the ratio of the number of the second samples SP2 for each measured volume magnetic susceptibility.

[0128] As shown in Table 1 and FIG. 7, the first sample SP1 and the second sample SP2 have different volume magnetic susceptibilities. The difference in volume magnetic susceptibility is due to the fact that d-limonene is encapsulated in the microcapsule D12. Specifically, the volume magnetic susceptibility of d-limonene is −8.48×10 -6 " As shown in Table 1, the average value of the volume magnetic susceptibility of the second sample SP2 (microcapsules D12 after washing) is closer to the volume magnetic susceptibility of d-limonene than the average value of the volume magnetic susceptibility of the first sample SP1 (yeast capsule substrate D11 after washing). Also, as shown in FIG. 7, the peak of the volume magnetic susceptibility of the second sample SP2 (microcapsules D12 after washing) is closer to the volume magnetic susceptibility of d-limonene than the peak of the volume magnetic susceptibility of the first sample SP1 (yeast capsule substrate D11 after washing). Therefore, it was possible to quantitatively analyze that d-limonene is encapsulated in the microcapsules D12.

[0129] [Example 2] In Example 2, similarly to Example 1, the volume magnetic susceptibility of the first sample SP11 (yeast capsule substrate D21 after washing) and the volume magnetic susceptibility of the second sample SP12 (microcapsule D22 after washing) were measured. Example 2 is similar to Example 1 except that the type of yeast capsule substrate is different. In detail, in Example 2, after a paste-like solid component is obtained from baker's yeast by a hot water treatment method, an emulsifier is added to the paste-like solid component. Then, a protease is further added to the paste-like solid component. A glycerin fatty acid ester is used as the emulsifier. An endo-type protease is used as the protease. The yeast capsule substrate D21 has a hydrophobic region inside.

[0130] Table 2 shows the measurement results of Example 2. [Table 2]

[0131] Fig. 8 is a diagram showing the measurement results according to Example 2. In detail, Fig. 8 shows the measurement results of the volume magnetic susceptibility of the first sample SP11 and the second sample SP12. In Fig. 8, the horizontal axis shows the volume magnetic susceptibility, and the vertical axis shows the percentage (%) of the particle number. In Fig. 8, the solid line graph shows the measurement results of the volume magnetic susceptibility of the first sample SP11 (yeast capsule base material D21 after washing), and the dashed line graph shows the measurement results of the volume magnetic susceptibility of the second sample SP12 (microcapsules D22 after washing).

[0132] As shown in Table 2 and FIG. 8, the first sample SP11 and the second sample SP12 have different volume magnetic susceptibilities. Specifically, as shown in Table 2, the average value of the volume magnetic susceptibility of the second sample SP12 (microcapsules D22 after washing) is closer to the volume magnetic susceptibility of d-limonene than the average value of the volume magnetic susceptibility of the first sample SP11 (yeast capsule substrate D21 after washing). Also, as shown in FIG. 8, the peak of the volume magnetic susceptibility of the second sample SP12 (microcapsules D22 after washing) is closer to the volume magnetic susceptibility of d-limonene than the peak of the volume magnetic susceptibility of the first sample SP11 (yeast capsule substrate D21 after washing). Therefore, it was possible to quantitatively analyze that d-limonene was encapsulated in the microcapsules D22.

[0133] [Example 3] In Example 3, similarly to Example 1, the volume magnetic susceptibility of the first sample SP21 (yeast capsule substrate D31 after washing) and the volume magnetic susceptibility of the second sample SP22 (microcapsule D32 after washing) were measured. Example 3 is similar to Example 1, except for the type of yeast and the process for releasing intracellular components from the yeast. More specifically, in Example 3, torula yeast was used. In addition, intracellular components were released from torula yeast by an enzymatic decomposition method. The yeast capsule substrate D31 has a hydrophobic region inside.

[0134] Table 3 shows the measurement results of Example 3. [Table 3]

[0135] Fig. 9 is a diagram showing the measurement results according to Example 3. Specifically, Fig. 9 shows the measurement results of the volume magnetic susceptibility of the first sample SP21 and the second sample SP22. In Fig. 9, the horizontal axis shows the volume magnetic susceptibility, and the vertical axis shows the percentage (%) of the particle number. In Fig. 9, the solid line graph shows the measurement results of the volume magnetic susceptibility of the first sample SP21 (yeast capsule base material D31 after washing), and the dashed line graph shows the measurement results of the volume magnetic susceptibility of the second sample SP22 (microcapsules D32 after washing).

[0136] As shown in Table 3 and FIG. 9, the first sample SP21 and the second sample SP22 have different volume magnetic susceptibilities. Specifically, as shown in Table 3, the average value of the volume magnetic susceptibility of the second sample SP22 (microcapsules D32 after washing) is closer to the volume magnetic susceptibility of d-limonene than the average value of the volume magnetic susceptibility of the first sample SP21 (yeast capsule substrate D31 after washing). Also, as shown in FIG. 9, the peak of the volume magnetic susceptibility of the second sample SP22 (microcapsules D32 after washing) is closer to the volume magnetic susceptibility of d-limonene than the peak of the volume magnetic susceptibility of the first sample SP21 (yeast capsule substrate D31 after washing). Therefore, it was possible to quantitatively analyze that d-limonene was encapsulated in the microcapsules D32.

[0137] [Example 4] In Example 4, the mixing ratio of the yeast capsule base material D21 in the form of a paste, d-limonene, and distilled water was changed to carry out the encapsulation process, and the volume magnetic susceptibility of the microcapsule D22 (sample SP3) after washing was measured for each mixing ratio. Specifically, the mixing ratio of the yeast capsule base material D21 in the form of a paste, d-limonene, and distilled water was changed in the range of "2:0:8" to "2:2.8:5.2" by weight ratio. Specifically, the mixing ratio of the yeast capsule base material D21, d-limonene, and distilled water was set to "2:0:8", "2:0.125:7.875", "2:0.25:7.75", "2:0.5:7.5", "2:1:7", "2:2:6", "2:2.5:5.5", and "2:2.8:5.2" by weight ratio.

[0138] Fig. 10 is a diagram showing a first measurement result according to Example 4. Specifically, Fig. 10 shows the measurement result of the volume magnetic susceptibility of sample SP3 (microcapsules D22 after washing). In Fig. 10, the horizontal axis shows the blending ratio (weight ratio) of d-limonene, and the vertical axis shows the volume magnetic susceptibility (average value).

[0139] As shown in Fig. 10, as the blending ratio of d-limonene increased, the volume magnetic susceptibility of sample SP3 (microcapsule D22 after washing) approached that of d-limonene. This result suggests that as the blending ratio of the component to be encapsulated increased, the amount of the component to be encapsulated in the microcapsule increased, and the volume magnetic susceptibility of the microcapsule changed.

[0140] In Example 4, further, the amount of d-limonene contained in sample SP3 (microcapsule D22 after washing) was measured using high performance liquid chromatography (HPLC). Specifically, sample SP3 was added to ethanol and stirred for 30 minutes using a vortex mixer to prepare solution L9. After stirring, solution L9 was centrifuged. Thereafter, the supernatant was measured using high performance liquid chromatography (HPLC). The HPLC conditions are shown below.

[0141] <HPLC Conditions> · High performance liquid chromatograph: manufactured by Hitachi High-Technologies Corporation · Column: Poroshell 120 EC-C18 manufactured by Agilent Technologies, Inc. (particle size; 2.7 μm, dimensions; 4.6 mm × 100 mm) · Column temperature: 40°C · Eluent: mixed solution of acetonitrile and water (blending ratio of acetonitrile and water is 50 wt%: 50 wt%) · Flow rate: 1 mL / min · Detector: ultraviolet light with a wavelength of 210 nm · Injection volume: 10 μL

[0142] Fig. 11 is a diagram showing the second measurement result according to Example 4. Specifically, Fig. 11 shows the measurement result of the amount of d-limonene contained in sample SP3. In Fig. 11, the horizontal axis indicates the blending ratio (weight ratio) of d-limonene, and the vertical axis indicates the amount of d-limonene contained in sample SP3. Specifically, the vertical axis indicates the percentage of the amount of d-limonene per 1 g of microcapsule.

[0143] 11, the amount of d-limonene contained in sample SP3 (microcapsules D22 after washing) increased as the blending ratio of d-limonene increased. Therefore, by increasing the blending ratio of d-limonene, the amount of d-limonene encapsulated in the microcapsules by the encapsulation treatment could be increased.

[0144] Fig. 12 is a diagram showing the third measurement result according to Example 4. Specifically, Fig. 12 shows the relationship between the amount of d-limonene contained in sample SP3 and the volume magnetic susceptibility of sample SP3. In Fig. 12, the horizontal axis shows the amount of d-limonene contained in sample SP3. Specifically, the horizontal axis shows the ratio of the amount of d-limonene per 1 g (unit weight) of microcapsules in percentage. The vertical axis shows the volume magnetic susceptibility (average value).

[0145] As shown in FIG. 12, in the range where the ratio of the amount of d-limonene per unit weight was 9% or less, as the amount of d-limonene per unit weight increased, the volume magnetic susceptibility of sample SP3 approached that of d-limonene. On the other hand, in the range where the ratio of the amount of d-limonene per unit weight exceeded 9%, as the amount of d-limonene per unit weight increased, the volume magnetic susceptibility of sample SP3 decreased. In other words, the volume magnetic susceptibility was saturated. From this result, it was confirmed that the amount of d-limonene encapsulated in the microcapsules could be quantitatively analyzed using the volume magnetic susceptibility until the volume magnetic susceptibility was saturated. Hereinafter, the amount of d-limonene encapsulated in the microcapsules will be referred to as the "amount of limonene encapsulated."

[0146] Fig. 13 is a diagram showing a fourth measurement result according to Example 4. In detail, Fig. 13 shows the relationship between the blending ratio of d-limonene and the ratio of the amount of encapsulated limonene to the total amount of d-limonene used in the encapsulation process. Hereinafter, the ratio of the amount of encapsulated limonene to the total amount of d-limonene used in the encapsulation process will be referred to as "ratio of encapsulated amount of limonene". In Fig. 13, the horizontal axis shows the blending ratio (weight ratio) of d-limonene, and the vertical axis shows the ratio (%) of the amount of encapsulated limonene.

[0147] As shown in FIG. 13, the amount of d-limonene contained in sample SP3 (microcapsule D22 after washing) increased as the blending ratio of d-limonene increased. On the other hand, the ratio of limonene encapsulation decreased as the blending ratio of d-limonene increased. This result suggests that the amount of the encapsulated component that is not encapsulated in the microcapsule increases as the blending ratio of the encapsulated component increases. Furthermore, even if the blending ratio of d-limonene is such that all of it is theoretically encapsulated in microcapsule D22, the ratio of encapsulated limonene did not reach 100%. This result suggests that the process of encapsulating the encapsulated component in the microcapsule follows the theory of chemical equilibrium.

[0148] [Example 5] In Example 5, salt was added to distilled water to carry out the encapsulation process, and the volume magnetic susceptibility of the microcapsule D22 (sample SP4) after washing was measured. In other words, the encapsulation process was carried out using saline as a solvent. The blending ratio of the paste-like yeast capsule base material D21, d-limonene, distilled water, and salt was 2:1:7:1 by weight.

[0149] Fig. 14 is a diagram showing the first measurement result according to Example 5. In detail, Fig. 14 shows the measurement result of the volume magnetic susceptibility of sample SP4. In Fig. 14, the horizontal axis shows the volume magnetic susceptibility, and the vertical axis shows the percentage (%) of the particle number. In Fig. 14, the solid line graph shows the measurement result of the volume magnetic susceptibility of sample SP4. Note that in Fig. 14, as a reference example, the dashed line graph shows the measurement result of the volume magnetic susceptibility of the second sample SP12 (microcapsules D22 after washing) described in Example 2.

[0150] As shown in FIG. 14, the addition of salt caused the volume magnetic susceptibility of the washed microcapsule D22 to become closer to the volume magnetic susceptibility of d-limonene.

[0151] In Example 5, similarly to Example 4, the amount of d-limonene contained in sample SP4 (the amount of d-limonene per 1 g of microcapsules) was measured using high performance liquid chromatography.

[0152] Fig. 15 is a diagram showing the second measurement result according to Example 5. Specifically, Fig. 15 shows the ratio of limonene encapsulation amount in sample SP4. Note that Fig. 15 also shows, as a reference example, the ratio of limonene encapsulation amount in the second sample SP12 (microcapsules D22 after washing) described in Example 2.

[0153] As shown in Figure 15, the amount of d-limonene encapsulated in microcapsules D22 after washing was increased by adding salt. Therefore, by adding salt to the solvent to reduce the solubility of d-limonene in the solvent, the amount of the component to be encapsulated in microcapsules D22 after washing was increased. This result suggests that the amount of the component to be encapsulated in the microcapsules is affected by the solubility of the component to be encapsulated in the solvent.

[0154] [Example 6] In Example 6, ethyl hexanoate was encapsulated in the yeast capsule substrate D21 as a component to be encapsulated, to prepare sample SP5 (microcapsule D22 after washing), and the volume magnetic susceptibility of sample SP5 was measured. In Example 6, the mixing ratio of the yeast capsule substrate D21 in the form of a paste, ethyl hexanoate, and distilled water was varied in the range of "2:0:8" to "2:2:6" by weight ratio to perform the encapsulation process. Specifically, the mixing ratio of the yeast capsule substrate D21, ethyl hexanoate, and distilled water was set to "2:0:8", "2:0.125:7.875", "2:0.25:7.75", "2:0.5:7.5", "2:1:7", and "2:2:6" by weight ratio. Here, the mixing ratios of ethyl hexanoate and distilled water respectively indicate the weights of ethyl hexanoate and distilled water when the weight of the yeast capsule substrate D11 is "2". Ethyl hexanoate is a hydrophobic substance.

[0155] Fig. 16 is a diagram showing the measurement results according to Example 6. Specifically, Fig. 16 shows the measurement results of the volume magnetic susceptibility of sample SP5. In Fig. 16, the horizontal axis shows the blending ratio (weight ratio) of ethyl hexanoate, and the vertical axis shows the volume magnetic susceptibility (average value).

[0156] As shown in Fig. 16, as the blending ratio of ethyl hexanoate increased, the volume magnetic susceptibility of sample SP5 (microcapsule D22 after washing) approached that of ethyl hexanoate. The volume magnetic susceptibility of ethyl hexanoate was -6.94 x 10 -6 This result suggests that as the blending ratio of the component to be encapsulated increases, the amount of the component to be encapsulated in the microcapsules increases, and the volume magnetic susceptibility of the microcapsules changes. [Industrial Applicability]

[0157] The present invention is useful, for example, in the evaluation of encapsulating materials for use in DDS. [Explanation of symbols]

[0158] 10 Analyzer 20 Magnetic field generation section 21 Cell 30 Observation Section 40 Arithmetic section 41 Storage section 42 Processing section

Claims

1. a first observation step of magnetophoretically observing the first particles; a first migration velocity measurement step of measuring a magnetophoretic velocity of the first particles from a result of the observation in the first observation step; a first magnetic susceptibility measurement step of measuring a volume magnetic susceptibility of the first particles based on a result of the measurement in the first migration velocity measurement step; an inclusion process for performing an inclusion process to cause the first particles to include a component to be included that is smaller than the first particles; a second observation step of magnetically phoresing the second particles to be analyzed, which are the first particles after the encapsulation treatment, and observing the second particles; a second migration velocity measuring step of measuring a magnetophoretic velocity of the second particles from a result of the observation in the second observation step; a second magnetic susceptibility measurement step of measuring a volume magnetic susceptibility of the second particles based on a result of the measurement in the second migration velocity measurement step; and analyzing whether or not the component to be included is included in the second particle based on a result of the measurement in the first magnetic susceptibility measurement step and a result of the measurement in the second magnetic susceptibility measurement step, a first zeta potential measurement step of measuring a zeta potential of the first particles; a second zeta potential measurement step of measuring the zeta potential of the second particles; and analyzing whether or not the component to be included is included in the second particle based on a result of the measurement in the first zeta potential measurement step and a result of the measurement in the second zeta potential measurement step. determining that the component to be encapsulated is encapsulated in the second particle when the volume magnetic susceptibility of the second particle is closer to the volume magnetic susceptibility of the component to be encapsulated compared to the volume magnetic susceptibility of the first particle; A particle analysis method, wherein the volume magnetic susceptibility of the component to be encapsulated is stored in advance in a memory unit provided in the analysis device.

2. The particle analysis method according to claim 1 , further comprising the step of spectroscopically observing the second particles to analyze whether or not the component to be included is included in the second particles.

3. A particle manufacturing method, comprising the particle analysis method according to claim 1 or 2.

4. The method for producing particles according to claim 3 , wherein the first particles contain solid components of the yeast that remain after intracellular components are released from the yeast.

5. The method further includes a first particle production step of producing the first particles, The particle production method according to claim 4 , wherein the first particle production step includes a step of releasing the intracellular components from the yeast.

6. The particle production method according to claim 3 , wherein the encapsulation process includes a process of mixing the first particles and the component to be encapsulated in a solvent.

Images