Composition, method for producing the composition, raw materials, penetrating composition, and penetration enhancer

Abstract

This invention provides a composition that improves the permeability of fucoxanthin into cell tissue. [Solution] The composition of the present invention comprises at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, characterized in that the weight ratio of the at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.0.

Description

Technical Field

[0001] The present invention relates to a composition, a method for producing the composition, a raw material body, a permeable composition, and a penetration enhancer.

Background Art

[0002] Fucoxanthin (C 42 H 58 O6), which is a red pigment contained in large amounts in brown algae such as wakame and kombu, has various excellent physiological activity functions such as anti-obesity, anti-diabetic, anti-inflammatory effects, and even whitening effects in addition to excellent antioxidant effects, and its use as a functional food material is eagerly desired. For this reason, various technologies related to various compositions containing fucoxanthin have been studied.

[0003] For example, Patent Document 1 discloses a process for producing fucoxanthin and / or polysaccharide from microalgae, and also discloses that it may contain an absorption promoter. In addition, Patent Document 2 discloses a composition containing carotenoid and fatty acid extracted from microalgae, and discloses that the fucoxanthin content is 1.7 to 2.0% / DW and the linoleic acid content is 0.23% DW.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0005] However, Patent Document 1 does not disclose what kind of substance is suitable as a fucoxanthin penetration enhancer, and Patent Document 2 also does not consider the permeability of fucoxanthin.

[0006] This invention has been made in view of the above-mentioned problems, and aims to provide a composition in which the permeability of fucoxanthin into cell tissue is improved, and a method for producing the same. [Means for solving the problem]

[0007] The present invention comprises at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, characterized in that the weight ratio of the at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.0.

[0008] The composition of the present invention comprises at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, and the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) to linoleic acid or a salt thereof is 1.1 to 22.0. Linoleic acid or its salts readily penetrate into cellular tissues such as skin, and it is hypothesized that at least one compound selected from the group consisting of fucoxanthin and its derivatives also penetrates into cellular tissues such as skin, induced by the penetration of linoleic acid or its salts. If the amount of at least one compound selected from the group consisting of fucoxanthin and its derivatives is too large, the presumed penetration effect of linoleic acid or its salts into cellular tissues relative to fucoxanthin may not be fully exerted. Conversely, if there is too much linoleic acid or its salts, it may preferentially penetrate into cellular tissues, making it difficult for fucoxanthin and other compounds to penetrate. For this reason, the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or its salts is considered to be between 1.1 and 22.0.

[0009] Examples of fucoxanthin derivatives include hydrolyzates or derivatives thereof of fucoxanthin, esters of fucoxanthin (e.g., esters with amino acids, carboxylic acids, inorganic acids, or fatty acids), salts thereof, and fucoxanthin glycosides. More specifically, derivatives of fucoxanthin include, but are not limited to, fucoxanthinol, a hydrolysis product; amarauciaxanthin A, obtained by dehydrating and isomerizing fucoxanthinol; esters of fucoxanthin with amino acids such as glycine and alanine; esters of fucoxanthin with carboxylic acids such as acetic acid and citric acid and their salts; esters of fucoxanthin with inorganic acids such as phosphoric acid and sulfuric acid and their salts; or monoesters and diesters of the same or different types of fucoxanthin selected from fatty acid esters of highly unsaturated fatty acids such as eicosapentaenoic acid and docosahexaenoic acid, unsaturated fatty acids such as oleic acid and linoleic acid, and saturated fatty acids such as palmitic acid and stearic acid, as well as glycosides such as glucosides.

[0010] Furthermore, metal salts and ammonium salts can be used as linoleic acid salts. In addition, alkali metal salts and alkaline earth metal salts can be used as metal salts, with lithium, sodium, and potassium being used as alkali metals, and calcium and magnesium being used as alkaline earth metals.

[0011] The composition of the present invention preferably excludes microalgae extracts. This is because microalgae extracts often contain unanalyzed proteins and impurities, which may cause allergic reactions or have an unpleasant odor, making them undesirable. Microalgae grow in either marine or freshwater environments. Microalgae are single-celled species that exist individually, in chains, or in groups and perform photosynthesis. Microalgae extract refers to a composition extracted from microalgae using a solvent. The solvents used include aqueous solvents, organic solvents, supercritical fluids, and subcritical fluids. Furthermore, for similar reasons, it is desirable that the composition of the present invention does not contain any components derived from microalgae.

[0012] In this invention, it is desirable that at least one compound selected from the group consisting of fucoxanthin and its derivatives, as well as linoleic acid or its salt, be derived from seaweed. This is because naturally derived compositions have fewer adverse effects on living organisms. Furthermore, the composition of the present invention may be a composition obtained by adding at least one compound selected from the group consisting of fucoxanthin derived from seaweed and its derivatives, and linoleic acid or a salt thereof, to industrially synthesized fucoxanthin and its derivatives, and linoleic acid or a salt thereof, thereby adjusting the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof. Furthermore, the term "seaweed" in this invention refers to a group of marine species of multicellular algae, and does not include microalgae, which are single-celled algae.

[0013] In this invention, it is desirable that the seaweed described above be the gametophyte of a seaweed. Seaweed exhibits various forms throughout its life cycle, including sporophytes (thallus, discus, filamentous) and gynandrophytes (thallus, discus, filamentous). Gynandrophytes are desirable in this invention because they contain high levels of fucoxanthin and its derivatives, as well as linoleic acid and its salts.

[0014] In this invention, it is preferable that the seaweed is the female gametophyte of the seaweed. This is because the female gametophyte contains the highest amount of fucoxanthin and its derivatives among the various forms in the life cycle of seaweed.

[0015] In this invention, the seaweed is preferably a brown alga. Examples of seaweed include green algae, red algae, and brown algae, but brown algae are preferred because they contain a large amount of fucoxanthin and its derivatives.

[0016] The composition of the present invention preferably does not contain an extract of Phaeodactylum tricornutum. Phaeodactylum tricornutum is a microalga, and extracts from it tend to contain allergens and substances that cause unpleasant odors. The extraction of Phaeodactylum tricornutum described above is carried out in accordance with Patent Document No. 2018-512432. The solvents used are methanol, ethanol, and supercritical carbon dioxide.

[0017] In the present invention, the composition may further contain polysaccharides.

[0018] In the present invention, the above composition comprises an organic solvent, or a mixed solvent consisting of an aqueous solvent and an organic solvent, wherein the organic solvent preferably dissolves at least one compound selected from the group consisting of fucoxanthin and its derivatives, and the organic solvent preferably is an organic solvent that dissolves at least one compound selected from the group consisting of fucoxanthin and its derivatives. In the present invention, the composition containing fucoxanthin, its derivatives, linoleic acid and its salts is easily soluble in an organic solvent. In the composition of the present invention, in addition to the organic solvent, it may further contain an aqueous solvent. The aqueous solvent is water or a solvent in which water-soluble salts are dissolved in water, and includes pure water, seawater, etc. Antifoaming agents, thixotropic agents, pH adjusters, etc. may be appropriately added to the aqueous solvent. Also, artificial seawater, filtered seawater, deep seawater, etc. can be used as the seawater. Also, in the present invention, the above composition may contain a mixed solvent composed of an aqueous solvent and an organic solvent. The above organic solvent preferably dissolves at least one compound selected from the group consisting of fucoxanthin and its derivatives.

[0019] In the present invention, the composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives and linoleic acid or its salt is preferably a powdery composition. Since the powdery composition can reduce the volume per component compared to the composition containing a solvent, the transportation cost can be reduced. Also, when used as a cosmetic, pharmaceutical, or food raw material, it is easy to mix with other raw materials and is useful as an industrial raw material.

[0020] The present invention is a method for producing a composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives and linoleic acid or its salt, which includes the following steps (1) to (3). Step (1): Disperse the gametophyte of seaweed in an aqueous solvent to obtain a dispersion of the gametophyte of the above seaweed. Step (2): Separate and remove the aqueous solvent from the dispersion of the gametophyte of the seaweed obtained in step (1). Step (3): Add an organic solvent to the gametophyte of the seaweed from which the aqueous solvent has been separated and removed in step (2), which dissolves at least one compound selected from the group consisting of fucoxanthin and its derivatives, to obtain a composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives and linoleic acid or a salt thereof, wherein the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.0.

[0021] Furthermore, the present invention is a method for producing a composition comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, comprising the following steps (1) to (4). Step (1): Disperse the gametophyte of the seaweed in an aqueous solvent to obtain a dispersion of the gametophyte of the seaweed. Step (2): The aqueous solvent is separated and removed from the dispersion of the gametophyte of the seaweed obtained in Step (1). Step (3): An organic solvent is added to the gametophyte of the seaweed from which the aqueous solvent was separated and removed in step (2) to obtain an organic solvent composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof. Step (4): Add at least one compound selected from the group consisting of fucoxanthin and its derivatives and / or linoleic acid or a salt thereof to the organic solvent composition obtained in Step (3), and adjust the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) to 1.1 to 22.0 to obtain a composition.

[0022] The gametophyte of seaweed is dispersed in an aqueous solvent, and the aqueous solvent is separated and removed from the gametophyte. Then, an organic solvent is added to the gametophyte of seaweed that dissolves at least one compound selected from the group consisting of fucoxanthin and its derivatives to obtain a composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives and linoleic acid or a salt thereof. If the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof in this composition is in the range of 1.1 to 22.0, there is no need to adjust the composition. However, if the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof is outside the range of 1.1 to 22.0, then at least one compound selected from the group consisting of fucoxanthin and its derivatives, or linoleic acid or a salt thereof is added to adjust the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof to 1.1 to 22.0. When adding at least one compound selected from the group consisting of fucoxanthin and its derivatives, or linoleic acid or a salt thereof, commercially available industrially synthesized products may be used. Furthermore, the amount of fucoxanthin and linoleic acid extracted increases if the gametophyte of the seaweed is first dispersed in an aqueous solvent, and then an organic solvent containing at least one compound selected from the group consisting of fucoxanthin and its derivatives is added. The reason for this is not clear, but it is presumed that first dispersing the gametophyte of the seaweed in an aqueous solvent makes it easier for the organic solvent to penetrate into the cells.

[0023] The organic solvents used in the present invention may include alcohols such as methanol, ethanol, propanol, isopropanol, n-butanol, and butanediols (1,3-butanediol (1,3-butylene glycol), 1,4-butanediol (1,4-butylene glycol)), ketones such as methyl ethyl ketone and acetone, esters such as methyl acetate and ethyl acetate, organochlorine hydrocarbons such as chloroform, aliphatic hydrocarbons such as hexane, and aromatic hydrocarbons such as benzene and toluene.

[0024] The composition of the present invention may contain an aqueous solvent in addition to the organic solvent. The aqueous solvent is water or a solvent in which water-soluble salts are dissolved in water, and includes pure water and seawater. An antifoaming agent, a thixotropic agent, a pH adjuster, etc., may be added to the aqueous solvent as appropriate. Artificial seawater, filtered seawater, deep-sea water, etc., can also be used as seawater. Examples of salts include sodium bicarbonate, potassium bicarbonate, sodium carbonate, potassium carbonate, ammonium sulfate, potassium phosphate, potassium dihydrogen phosphate, iron phosphate, iron fulvicate, and sodium citrate. The aqueous solvent may also be phosphate-buffered saline.

[0025] The weight ratio of the organic solvent to the aqueous solvent should preferably be organic solvent / aqueous solvent = 1 / 10 to 10 / 10.

[0026] The method for producing the composition of the present invention may further include a drying step of drying the above composition to make it into a powder.

[0027] The present invention relates to a raw material used for extracting and preparing a composition comprising an aggregate of gametophytes of seaweed, containing at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, wherein the weight ratio of the at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.0.

[0028] This is because the gametophytes of seaweed contain high levels of at least one compound selected from the group consisting of fucoxanthin and its derivatives, as well as linoleic acid or its salts.

[0029] The seaweed mentioned above should preferably be brown algae, as brown algae contain the highest levels of fucoxanthin and its derivatives among all types of seaweed.

[0030] The above-mentioned seaweed is preferably female gametophyte, because it contains more fucoxanthin and its derivatives than male gametophyte.

[0031] The cell tissue permeable composition of the present invention comprises at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, characterized in that the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or salt thereof]) is 1.1 to 22.0.

[0032] The cell tissue penetration enhancer of the present invention comprises at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, characterized in that the weight ratio of the at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.0.

[0033] By using the permeable composition and permeation enhancer of the present invention, fucoxanthin can be suitably permeated into cell tissue. [Effects of the Invention]

[0034] According to the present invention, it is possible to provide a composition in which the permeability of fucoxanthin into cell tissue is improved. By using the composition of the present invention, at least one compound selected from the group consisting of fucoxanthin and its derivatives penetrates into the cell tissue through the action of linoleic acid or its salt, thereby imparting various excellent physiological functions to the cell tissue, such as the excellent antioxidant effect, anti-obesity effect, anti-diabetic effect, anti-inflammatory effect, and even a whitening effect of fucoxanthin. For this reason, the composition of the present invention has a wide range of applications, including pharmaceuticals, food products, and cosmetics. [Brief explanation of the drawing]

[0035] [Figure 1] Figure 1 is a schematic diagram showing the female and male gametophytes. [Figure 2] Figure 2 is a liquid chromatography chart of fucoxanthin in an extract obtained by water extraction of the female gametophyte of wakame seaweed, followed by further extraction with 100% butylene glycol. In Figure 2, the peak around 13 minutes corresponds to fucoxanthin. The peak around 11 minutes in Figure 2 corresponds to chlorophyll a. [Figure 3A]Figure 3A is a chart showing the analysis results of linoleic acid by LC / MS with time (min) on the horizontal axis. The peak around 4.2 minutes is the peak of linoleic acid. In Figure 3A, there appear to be four peaks from 4.17 to 4.31 minutes, but this is because a single linoleic acid is being measured, and a calibration curve can be created from the area of ​​the peak around 4.2 minutes. The detector used is the Q Exactive Focus, manufactured by Thermo Fisher Scientific Co., Ltd., which has the function of alternately measuring negatively charged molecular ions and positively charged molecular ions (positive-negative alternating scan). Linoleic acid molecular ions have a negative charge, and when negatively charged molecular ions are measured for a predetermined time, a part of the linoleic acid peak appears on the chart. After the predetermined time has elapsed, the instrument then starts measuring positively charged molecular ions, so the rise of the linoleic acid molecular ion peak stops, and a shoulder appears on the peak. When positive charges are measured for a predetermined period, the system begins measuring negatively charged molecular ions again, causing the linoleic acid peak to rise once more. In this way, because positively and negatively charged molecular ions are measured alternately, a shoulder appears on the linoleic acid peak, which has a negative charge. However, what is being measured is the negatively charged molecular ion of linoleic acid. [Figure 3B] Figure 3B is a chart showing the results of LC / MS analysis of linoleic acid with the mass-to-charge ratio (m / z) on the horizontal axis. The peak at m / z = 279.23 in Figure 3B is the peak of the molecular ion of linoleic acid. [Figure 4] Figure 4 shows the results of the tape stripping test. The vertical axis represents the penetration rate, and the horizontal axis represents the weight ratio of fucoxanthin to linoleic acid. [Modes for carrying out the invention]

[0036] The present invention comprises at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, wherein the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.0. Linoleic acid or its salts readily penetrate into cell tissues, and it is hypothesized that at least one compound selected from the group consisting of fucoxanthin and its derivatives may also penetrate into cell tissues, induced by the penetration of linoleic acid or its salts. However, if the amount of at least one compound selected from the group consisting of fucoxanthin and its derivatives is too large, the presumed penetration effect of linoleic acid or its salts into cell tissues relative to fucoxanthin may not be fully exerted. Conversely, if there is too much linoleic acid or its salts, it may preferentially penetrate into cell tissues, making it difficult for fucoxanthin and other compounds to penetrate. For this reason, the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or its salts ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or its salt]) is considered to need to be between 1.1 and 22.0.

[0037] Cellular tissues include epithelial tissues such as the epidermis of the skin, the inner wall of the digestive tract, alveoli, and vascular endothelium; connective tissues such as bone, cartilage, blood, lymph, adipose tissue, tendons, and ligaments; muscle tissues such as skeletal muscle, cardiac muscle, and smooth muscle; and nervous tissues such as the brain, spinal cord, and nerves.

[0038] The composition of the present invention preferably excludes microalgae extracts. This is because microalgae extracts often contain unanalyzed proteins and impurities, which may cause allergic reactions or have an unpleasant odor, making them undesirable. Microalgae grow in either marine or freshwater environments. Microalgae are single-celled species that exist individually, in chains, or in groups and perform photosynthesis. Although the term "micro" is used, size itself is not the essential characteristic; single-celled algae are simply called microalgae. Microalgae extract refers to a composition extracted from microalgae using a solvent. The solvents used include aqueous solvents, organic solvents, supercritical fluids, and subcritical fluids. Furthermore, for similar reasons, it is desirable that the composition of the present invention does not contain any components derived from microalgae. Microalgae include Phaeodactylum tricornutum, Navicula pelliculosa, Amphora, Isochrysis aff. Galbana, Odontella aurita, Nitzscia closterium, Cylindrotheca closter ium, Chaetoseros sp., Emiliania huxleyi, Phaeodactykum sp., Iso chrysis sp., Amphora sp., Naviculla lensi, Examples include Naviculla incerta and Chaeotocerous sp., Phaeodactylum tricornutum (P. tricornutum), and OPMS30543 strain (Pavlova).

[0039] Furthermore, it is desirable that the composition of the present invention does not contain an extract of Phaeodactylum tricornutum. Phaeodactylum tricornutum is a microalga, and extracts from it tend to contain allergens and substances that cause unpleasant odors.

[0040] Examples of fucoxanthin derivatives include hydrolysates or derivatives thereof of fucoxanthin, esters of fucoxanthin (e.g., esters with amino acids, carboxylic acids, inorganic acids, or fatty acids), salts thereof, and fucoxanthin glycosides. More specifically, examples of fucoxanthin derivatives include, but are not limited to, monoesters and diesters of the same or different types, glycosides, etc., selected from fucoxanthin derivatives such as fucoxanthin, fucoxanthin, fucoxanthin, fucoxanthin, ferro

[0041] The total content of at least one compound selected from the group consisting of fucoxanthin and its derivatives in the composition of the present invention is not particularly limited, but may be, for example, 0.0011 to 50% by weight in the composition. Similarly, the total content of linoleic acid or its salt in the composition of the present invention is not particularly limited, but may be, for example, 0.001 to 45% by weight in the composition.

[0042] Furthermore, seaweed can be used as a raw material for producing at least one compound selected from the group consisting of fucoxanthin and its derivatives, or linoleic acid or its salt, and brown algae, red algae, green algae, etc., can be used. In this invention, "seaweed" refers to multicellular algae, specifically marine species, and does not include microalgae, which are single-celled algae. Examples of green algae include *Ulva lactuca*, *Monostroma*, *Euphyllia*, *Euphyllia glabrescens*, *Euphyllia erythrosora*, and *Caulerpa lentillifera*. Examples of red algae include *Ulva perforata*, *Ulva serrata*, *Ulva perforata*, *Porphyra japonica*, *Porphyra japonica* (Japanese name: *Haitan Amanori*), *Ulva perforata*, *Porphyra japonica*, *Ulva perforata*, *Ulva perforata*, *Ulva perforata*, *Ulva perforata*, *Ulva perforata*, *Ulva perforata*, and *Ulva perforata*. Examples of brown algae include *Wakame*, *Mozuku* (Okinawa mozuku, Itomozuku), *Habanoori*, *Hirome*, *Aowakame*, *Sagarame*, *Kayamonori*, *Kelp*, *Makonbu*, *Hosomekomu*, *Nagakonbu*, *Mitsuishikonbu*, *Kajime*, *Kurome*, *Hondawara*, *Arame*, and *Seiyo Habanoori*. Among seaweeds, brown algae are the most desirable because they contain a high amount of fucoxanthin or its derivatives. Of the brown algae, wakame is the most preferred. Seaweed can take various forms, including sporophytes (thallus, discus, filamentous) and gynandrophytes (thallus, discus, filamentous). Gynandrophytes are desirable in this invention because they contain a high amount of fucoxanthin or its derivatives. Female gametophytes are particularly advantageous because they contain a higher amount of fucoxanthin or its derivatives and linoleic acid or its salts compared to male gametophytes.

[0043] The composition of the present invention may further contain polysaccharides.

[0044] The composition of the present invention preferably contains an organic solvent. As the organic solvent, any organic solvent capable of dissolving at least one compound selected from the group consisting of fucoxanthin and its derivatives can be used. Such organic solvents can be alcohols such as methanol, ethanol, propanol, isopropanol, n-butanol, and butanediol (1,3-butanediol (1,3-butylene glycol), 1,4-butanediol (1,4-butylene glycol)), ketones such as methyl ethyl ketone and acetone, esters such as methyl acetate and ethyl acetate, organochlorine hydrocarbons such as chloroform, aliphatic hydrocarbons such as hexane, and aromatic hydrocarbons such as benzene and toluene, either alone or in combination of two or more. Among these, ethanol and butanediol (butylene glycol) are preferred as organic solvents.

[0045] In this invention, an aqueous solvent may be included in addition to the organic solvent. The aqueous solvent mentioned above is water or a solvent in which water-soluble salts are dissolved, and includes pure water and seawater. Antifoaming agents, thixotropic agents, pH adjusters, etc., may be added to the aqueous solvent as appropriate. Artificial seawater, filtered seawater, or deep-sea water can also be used as seawater. Examples of salts include sodium bicarbonate, potassium bicarbonate, sodium carbonate, potassium carbonate, ammonium sulfate, potassium phosphate, potassium dihydrogen phosphate, iron phosphate, iron fulvicate, and sodium citrate. The aqueous solvent may also be phosphate-buffered saline. The composition of the present invention may also contain an organic solvent in addition to the aqueous solvent. The organic solvent is preferably one that dissolves at least one compound selected from the group consisting of fucoxanthin and its derivatives. The weight ratio of the organic solvent to the aqueous solvent should preferably be organic solvent / aqueous solvent = 1 / 10 to 10 / 10.

[0046] In one embodiment of the present invention, the composition comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, is preferably a powder composition. Since powder compositions have a smaller volume per component compared to compositions containing solvents, transportation costs can be reduced. Furthermore, when used as a raw material for cosmetics, pharmaceuticals, or food products, they are easily mixed with other raw materials and are useful as industrial raw materials.

[0047] The composition of the present invention may contain components other than those described above, as long as they do not impair the effects of the present invention. Examples of such components include excipients and additives that can be used in cosmetics, pharmaceuticals, foods, and their raw materials.

[0048] Next, a method for producing a composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, will be described.

[0049] The present invention provides a method for producing a composition comprising, for example, a gametophyte of seaweed as a raw material, and the following steps (1) to (3): (1) a compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof.

[0050] Step (1): Disperse the gametophytes of seaweed in an aqueous solvent to obtain a dispersion of seaweed gametophytes. Step (2): The aqueous solvent is separated and removed from the dispersion of seaweed gametophytes obtained in Step (1). Step (3): Add an organic solvent to the gametophyte of the seaweed from which the aqueous solvent was separated and removed in step (2) to obtain a composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives and linoleic acid or a salt thereof, wherein the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.0.

[0051] Another aspect of the method for producing the composition of the present invention is, for example, a method for producing a composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, using the gametophyte of seaweed as a raw material, and comprising the following steps (1) to (4).

[0052] Step (1): Disperse the gametophytes of seaweed in an aqueous solvent to obtain a dispersion of seaweed gametophytes. Step (2): The aqueous solvent is separated and removed from the dispersion of seaweed gametophytes obtained in Step (1). Step (3): An organic solvent is added to the gametophyte of the seaweed from which the aqueous solvent was separated and removed in step (2) to obtain an organic solvent composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof. Step (4): Add at least one compound selected from the group consisting of fucoxanthin and its derivatives and / or linoleic acid or a salt thereof to the organic solvent composition obtained in Step (3), and adjust the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) to 1.1 to 22.0 to obtain a composition.

[0053] The gametophyte of the seaweed used in the method for producing the composition of the present invention described above is preferably a female gametophyte. Examples of seaweed include the seaweeds mentioned above, preferably brown algae, and preferably wakame seaweed.

[0054] The following method will be explained using the female gametophyte of seaweed as an example, but it is not limited to this. First, we will explain one example of how to obtain the gametophyte of seaweed, using wakame as an example.

[0055] (I) Development of gametophytes of seaweed The gametophytes that develop from the zoospores of wakame seaweed are separated into one male and one female, propagated, mechanically shredded, attached to or adhering to threads, and fertilized on the threads. The young wakame leaves that emerge from this stage will be grown in indoor culture and in natural seawater. From the mature wakame thallus (adult), spore-bearing leaves (mekabu) will be obtained.

[0056] 1. Preservation of spore-bearing fronds 1) Place the spore-bearing fronds in a plastic bag or container and store them in a cool, dark place at 15-20°C. Below 15°C, zoospores will not be easily released when returned to seawater. If the spore-bearing fronds are removed from seawater and stored in a cool, dark place, zoospore release is possible for 2-3 days.

[0057] 2. Collection of zoospores 1) The ideal room temperature for releasing zoospores is 15-20°C. Higher temperatures are undesirable because they shorten the zoospore's swimming time.

[0058] 2) Cut the spore-bearing fronds into pieces about 3-4 cm square. The part closest to the rhizoids releases spores well, but cut off the part of the spore-bearing frond surface that is as clean as possible. Gently wipe off any dirt from the cut leaf pieces with absorbent paper or similar.

[0059] 3) Prepare three beakers containing 100 mL of sterile seawater, wash the leaf fragments one by one, and then place them in petri dishes containing 50 mL of sterile seawater.

[0060] 4) Place the petri dish containing the leaf fragments on the stereomicroscope stage and shine light from above the petri dish to induce zoospore release. Adjusting the stereomicroscope to a dark-field setting makes it easier to observe zoospore release. After about 10 minutes of illumination using an optical fiber or similar device, sufficient zoospores will be released.

[0061] 5) Prepare a capillary tube. Use either a heated and stretched hematocrit tube, or a Pasteur pipette tip that has been sufficiently stretched to a long, thin shape.

[0062] 6) Prepare a petri dish filled with 50 mL of PESI culture medium.

[0063] 7) Aspirate an appropriate amount of zoospores under a stereomicroscope and drop them into a petri dish. Take care to prevent the capillary tube from touching the bottom of the petri dish or leaf fragments during aspiration (diatoms are often aspirated). After dropping, shake the petri dish thoroughly by hand to equalize the zoospore density.

[0064] 8) Prepare about four different petri dishes, each with a different amount of zoospore fluid aspirated. A large amount of zoospores will increase the density of the gametophytes, causing them to grow too close together and making isolation difficult.

[0065] 9) After collecting zoospores, the petri dishes should be cultured at 10°C to 30°C with 14 to 12 hours of light (1000 to 1500 lux). Since the gametophytes can become similar in shape to males and females and be difficult to distinguish if there are large temperature fluctuations, high temperatures, or high light levels, care should be taken to maintain homeostasis in the culture conditions.

[0066] 10) The gametophytes will reach a size where their sex can be determined in two weeks. Due to the risk of fertilization, the sex of the gametophytes should be determined and isolated as soon as possible.

[0067] 11) At this stage, there is little to no contamination by diatoms, but if diatoms appear, discard the petri dish. If discarding is not possible, germanium dioxide can be used to suppress diatom growth, and the gametophyte that is not contaminated by diatoms can be isolated.

[0068] 3. Isolation of male and female gametophytes 1) Place the petri dish containing the cultured gametophytes on the stage of the inverted microscope and search for female gametophytes suitable for isolation. Isolate those gametophytes that are sufficiently separated from each other and whose sex is clearly defined. Attach a tube to a Pasteur pipette and separate the female gametophytes from the petri dish and aspirate them. Place the aspirated female gametophytes one by one into microplates filled with PESI medium. Schematic diagrams of female and male gametophytes are shown in Figure 1. 2) Cultivate for one month at 10-30°C with 14-12 hours of light (1500-2000 lux).

[0069] 4. Preservation of the female gametophyte 1) Remove the female gametophytes from the microplate after culturing. Normally, the female gametophytes are large enough to be seen with the naked eye, so pick them up from the microplate with ophthalmic forceps. If they are not large enough, you can use a Pasteur pipette to aspirate them under an inverted microscope. 2) When storing, place in a screw-cap test tube and store at 10°C to 30°C with 14 hours of light (1000 to 1500 lux). After storage, the PESI medium is replaced once every two months.

[0070] (II) Cultivation of gametophytes of seaweed For the culture medium of the female gametophyte, Provasoli's Enriched Seawater (PES) or its improved form, PESI culture medium, is preferable. The PES culture medium has the following composition. (See [Hirokazu Ariga, Isao Inoue, Jiro Tanaka, Yasutsugu Yokohama, and Tadao Yoshida, eds., "Phycology: Experiments and Practical Training," Kodansha Scientific (2000)] and Japanese Patent Publication No. 2009-201480.) Furthermore, PESI culture media can be prepared by referring to Plant Tissue Culture, 6(2), 55-62 (1989), etc. The detailed composition will be described later in the examples.

[0071] (PES culture solution) Tris hydroxymethyl aminomethane 5.0g NaNO33.5g Na2-glycerophosphate 500mg Fe stock solution 250mL P-2 metal mix 250mL Vitamine B12 stock solution (0.1mg / mL) 1.0mL Thiamine-HCl stock solution (1.0mg / mL) 5.0mL Biotine stock solution (0.1mg / mL) 0.5mL Distilled water 1000mL

[0072] (Fe stock solution) Na2-EDTA·2H2O 330mg Fe(NH4)2(SO4)2·6H2O 351mg Distilled water 500mL

[0073] (P-2 metal mix) Na2-EDTA·2H2O 500mg H3BO 3570mg FeCl3·6H2O 24.5mg MnSO4·4H2O 82.0mg CoSO4·7H2O (4.8 mg / mL) 0.5 mL ZnSO4·7H2O 11.0mg Distilled water 500mL

[0074] A culture solution for seaweed is prepared by adding PES or PESI culture solution to seawater. The seawater used may be natural or artificial seawater that has been sterilized by ozone, ultraviolet light, or autoclave, or filtered seawater obtained by filtering natural seawater may be used. Artificial seawater is prepared to mainly contain cations such as sodium ions, magnesium ions, potassium ions, and calcium ions, and anions such as chloride ions and sulfate ions. The salinity of the artificial seawater should preferably be between 1.0% by mass and 3.5% by mass. When using natural seawater, deep-sea water may also be used. Deep-sea water is seawater distributed in the deep sea at depths of 200m or more. Because deep-sea water has a high salt concentration, bacteria and other organisms that hinder seaweed cultivation are less likely to survive. In addition, deep-sea water is free from artificial pollution, has low temperatures resulting in fewer bacteria, and is free from the presence of phytoplankton due to the absence of sunlight. Furthermore, deep-sea water is rich in nitrogen (N) in the form of nitrates, phosphorus (P) in the form of phosphates, and silicon (Si) in the form of silicates, making it suitable for seaweed growth.

[0075] Female gametophytes, which are seaweed, are placed in the prepared seaweed culture medium, and aeration culture is performed. Aeration culture consists of a preliminary culture and a main culture. The female gametophytes of seaweed are cultured in an artificial environment, which includes an environment in which water temperature, light intensity, and daylight hours are controlled. Any environment is acceptable as long as the light intensity, daylight hours, and water temperature can be precisely controlled. For example, seaweed may be cultured in a container that allows cultivation within an incubator where the light intensity, daylight hours, and internal temperature are controlled, or it may be cultured in a tank in an environment where the light intensity, daylight hours, and water temperature can be controlled.

[0076] In the preliminary culture environment, a daylight-white fluorescent lamp is used as the light source, with an irradiation intensity of 1-100 μmol / m². 2 The ideal conditions are a water temperature of 5-30°C and 8-24 hours of daylight. Cultivation is usually carried out under alternating light and dark periods, but continuous light irradiation is also acceptable. The cultivation period can be between 1 and 40 days, but preferably 5-10 days. Even a minimum of 5 days is sufficient.

[0077] In this culture environment, a green light source (490-550 nm) is used, and the irradiation intensity is 10-200 μmol / m². 2 The ideal conditions are a water temperature of 5-30°C and 8-24 hours of daylight. Cultivation is usually carried out under alternating light and dark periods, but continuous light irradiation is also permitted. The cultivation period may be between 1 and 40 days, but 5-10 days is preferable. The culture solution for seaweed used in this invention may contain appropriate seaweed growth promoters such as fatty acids. The culture tank may also be equipped with one or more devices selected from a stirring device, a vibrating device, a temperature control device, a pH adjustment device, a turbidity measuring device, a light control device, a device for measuring the concentration of specific gases such as air, O2, and CO2, and a pressure measuring device. Seaweed cultivation can be carried out using any suitable liquid culture method, such as batch culture, semi-batch culture (fed-batch culture), or continuous culture (perfusion culture).

[0078] After culturing the female gametophytes of seaweed, they are either scooped up with a plankton net or filtered to extract the gametophytes from the culture medium. The aggregate of seaweed gametophytes obtained in this way can be preferably used as a raw material for extracting, for example, at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or its salt.

[0079] (III) Aqueous solvent dispersion and extraction of gametophytes of seaweed The gametophyte of the seaweed is dispersed in an aqueous solvent to obtain a dispersion of the gametophyte of the seaweed (step (1)). By dispersing the gametophyte of the seaweed in an aqueous solvent, it is also possible to extract some of fucoxanthin and its derivatives from the gametophyte into the aqueous solvent. If necessary, the female gametophyte of the seaweed is dispersed in an aqueous solvent, and some of at least one compound selected from the group consisting of fucoxanthin and its derivatives is extracted from the female gametophyte of the seaweed into the dispersion. Furthermore, in the case of seaweed gametophytes, it is easier to extract at least one compound selected from the group consisting of fucoxanthin and its derivatives, as well as linoleic acid or its salts, by first dispersing the seaweed gametophytes in an aqueous solvent, then separating and removing the aqueous solvent, and finally extracting the seaweed gametophytes with an organic solvent, rather than directly extracting with an organic solvent. An aqueous solvent is water or a solvent in which water-soluble salts are dissolved, and includes pure water and seawater. Antifoaming agents, thixotropic agents, pH adjusters, etc., may be added to the aqueous solvent as appropriate. Artificial seawater, filtered seawater, or deep-sea water can also be used as seawater. Examples of salts include sodium bicarbonate, potassium bicarbonate, sodium carbonate, potassium carbonate, ammonium sulfate, potassium phosphate, potassium dihydrogen phosphate, iron phosphate, iron fulvicate, and sodium citrate. Phosphate-buffered saline (PBS) may also be used as the aqueous solvent.

[0080] The extraction temperature should preferably be between 0°C and 60°C. To prevent degradation of fucoxanthin, it is even more desirable to extract while cooling with ice (cooling to 0°C). The extraction time should preferably be between 0.5 and 5 hours. It is desirable to disperse the gametophyte of the seaweed (preferably the female gametophyte) in an aqueous solvent and stir or permeate it. It is desirable to use 0.01 to 1 L of aqueous solvent per 1 g of dry weight of the seaweed gametophyte. The gametophytes of the seaweed are separated and removed from the resulting dispersion of gametophytes of the seaweed to obtain an aqueous solvent composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives (step (2)). In this specification, the aqueous solvent composition obtained in step (2) may also be referred to as an aqueous solvent extract, or as an aqueous solvent extract of the gametophytes or sporophytes of the seaweed. The extraction residue, including the gametophytes of seaweed and impurities, is removed from the aqueous solvent by methods such as filtration using filters or columns, or centrifugation. The gametophytes of the seaweed separated in step (2) can be used for extraction with organic solvents as described later.

[0081] (IV) Disperse the gametophytes of seaweed in an aqueous solvent, and separate the gametophytes from the aqueous solvent. It is desirable to add an organic solvent to the gametophyte of the seaweed separated in step (2) above to obtain an organic solvent composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives (step (3)). In this specification, the organic solvent composition obtained in step (3) may also be referred to as an organic solvent extract. The gametophyte of seaweed, which has been extracted with an aqueous solvent, is immersed in an organic solvent that dissolves at least one compound selected from the group consisting of fucoxanthin and its derivatives, thereby extracting at least one compound selected from the group consisting of fucoxanthin and its derivatives, as well as linoleic acid or a salt thereof, from the gametophyte of seaweed. The organic solvent causes at least one compound selected from the group consisting of fucoxanthin and its derivatives, as well as linoleic acid or its salt, to leach into the organic solvent from within the cells of the gametophyte of seaweed.

[0082] As the organic solvent to be used, an organic solvent that dissolves at least one compound selected from the group consisting of fucoxanthin and its derivatives can be used. As such organic solvents, alcohols such as methanol, ethanol, propanol, isopropanol, n-butanol, and butanediol (1,3-butanediol (1,3-butylene glycol), 1,4-butanediol (1,4-butylene glycol)), ketones such as methyl ethyl ketone and acetone, esters such as methyl acetate and ethyl acetate, organochlorine hydrocarbons such as chloroform, aliphatic hydrocarbons such as hexane, and aromatic hydrocarbons such as benzene and toluene can be used alone or in combination of two or more. Among these, ethanol and butanediol are preferred as organic solvents. At least one compound selected from the group consisting of fucoxanthin and its derivatives is eluted into the organic solvent. Since fucoxanthin and its derivatives are substantially insoluble in aqueous solvents, the content of fucoxanthin and its derivatives in the organic solvent is higher than that of the aqueous solvent extract in (II). The organic solvent may contain aqueous solvents, but it is preferable that it does not. If aqueous solvents are present in the organic solvent, they mainly originate from water contained in the gametophyte of seaweed. In extraction with organic solvents, the extraction temperature should preferably be between 0°C and 60°C. To prevent degradation of fucoxanthin and to increase the amount of fucoxanthin extracted, it is even more desirable to cool the extraction with ice (cooling to 0°C). The extraction time should preferably be between 0.5 and 5 hours. It is desirable to disperse the gametophyte of the seaweed in the organic solvent and stir or permeate it. It is desirable to use the organic solvent at a ratio of 0.01 to 1 L per 1 g of gametophyte of the seaweed. After extraction with an organic solvent, the extraction residue, which includes the gametophyte of the seaweed and impurities, may be removed from the organic solvent. The method for removing the gametophyte of the seaweed from the organic solvent is not particularly limited, and methods such as filtration using filters or columns, or centrifugation can be used. In this way, a composition (organic solvent composition) can be obtained that contains an organic solvent, at least one compound selected from the group consisting of fucoxanthin and its derivatives eluted in the organic solvent, and linoleic acid or a salt thereof.

[0083] (V) A composition is obtained by mixing an aqueous solvent extract of the seaweed gametophyte, an organic solvent extract, and at least one compound selected from the group consisting of fucoxanthin and its derivatives with linoleic acid or a salt thereof. If the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or its salt in this composition is in the range of 1.1 to 22.0, there is no need to adjust the composition. However, if the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or its salt is outside the range of 1.1 to 22.0, then at least one compound selected from the group consisting of fucoxanthin and its derivatives, or linoleic acid or its salt, should be added to adjust the weight ratio to linoleic acid or its salt to at least one compound selected from the group consisting of fucoxanthin and its derivatives to 1.1 to 22.0. When adding at least one compound selected from the group consisting of fucoxanthin and its derivatives, or linoleic acid or its salt, commercially available industrially synthesized products can be used.

[0084] In the method for producing the composition of the present invention, the above composition may be dried to form a powder. Various methods can be used to dry the composition, including heat drying, freeze-drying, and vacuum drying. In this case, the average particle size of the powder contained in the composition is preferably 0.1 to 10 μm.

[0085] The present invention provides a composition comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, wherein the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof is 1.1 to 22.0. This composition exhibits excellent permeability to cell tissues and can therefore be used in various industrial applications. In particular, it can be widely used in various health foods expected to have cholesterol-reducing and thrombosis-preventive effects, pharmaceuticals aimed at antitumor effects, neuroprotective effects, and blood glucose level elevation suppression effects, and cosmetics aimed at inhibiting melanin production.

[0086] As described above, the gametophyte of seaweed can be suitably used as a raw material for preparing a composition characterized by comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, wherein the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof is 1.1 to 22.0.

[0087] The present invention also includes raw materials used to extract and prepare a composition comprising an aggregate of gametophytes of seaweed, at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, wherein the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof is 1.1 to 22.0. The seaweed is preferably a brown alga. The seaweed is preferably a female gametophyte.

[0088] The present invention is also a cell tissue permeable composition comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, characterized in that the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof is 1.1 to 22.0.

[0089] Furthermore, the present invention is also a cell tissue penetration enhancer characterized by comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, wherein the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof is 1.1 to 22.0.

[0090] By using the permeable composition and permeation enhancer of the present invention, fucoxanthin can be suitably permeated into cell tissue.

[0091] This specification contains the following information:

[0092] The present disclosure (1) is a composition comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives and linoleic acid or a salt thereof, wherein the weight ratio of the at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.0.

[0093] Disclosure (2) is the composition according to Disclosure (1), comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, which is derived from seaweed.

[0094] Disclosure (3) is the composition described in Disclosure (2), wherein the seaweed is the gametophyte of the seaweed.

[0095] Disclosure (4) is the composition according to Disclosure (2) or (3), wherein the seaweed is the female gametophyte of the seaweed.

[0096] Disclosure (5) is a composition according to any one of Disclosures (2) to (4), wherein the seaweed is a brown alga.

[0097] Disclosure (6) is the composition according to any one of Disclosures (1) to (5) wherein the composition does not contain an extract of Phaeodactylum tricornutum.

[0098] Disclosure (7) is a composition according to any one of items (1) to (6) of this disclosure, further comprising a polysaccharide.

[0099] Disclosure (8) is a composition according to any one of items (1) to (7) of this disclosure, wherein the composition comprises an organic solvent or a mixed solvent consisting of an aqueous solvent and an organic solvent, and the organic solvent dissolves at least one compound selected from the group consisting of fucoxanthin and its derivatives.

[0100] Disclosure (9) states that the above composition is a powdered composition as described in any one of Disclosures (1) to (8).

[0101] This disclosure (10) is a method for producing a composition comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives and linoleic acid or a salt thereof, comprising the following steps (1) to (3). Step (1): Disperse the gametophyte of the seaweed in an aqueous solvent to obtain a dispersion of the gametophyte of the seaweed. Step (2): The aqueous solvent is separated and removed from the dispersion of the gametophyte of the seaweed obtained in Step (1). Step (3): Add an organic solvent to the gametophyte of the seaweed from which the aqueous solvent has been separated and removed in step (2) to obtain a composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives and linoleic acid or a salt thereof, wherein the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.0.

[0102] (11) This disclosure provides a method for producing a composition comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, comprising the following steps (1) to (4). Step (1): Disperse the gametophyte of the seaweed in an aqueous solvent to obtain a dispersion of the gametophyte of the seaweed. Step (2): The aqueous solvent is separated and removed from the dispersion of the gametophyte of the seaweed obtained in Step (1). Step (3): An organic solvent is added to the gametophyte of the seaweed from which the aqueous solvent was separated and removed in step (2) to obtain an organic solvent composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof. Step (4): Add at least one compound selected from the group consisting of fucoxanthin and its derivatives and / or linoleic acid or a salt thereof to the organic solvent composition obtained in Step (3), and adjust the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) to 1.1 to 22.0 to obtain a composition.

[0103] Disclosure (12) is a method for producing the composition according to Disclosure (10) or (11), further comprising a drying step of drying the composition to make it into a powder.

[0104] The present disclosure (13) is a raw material used for extracting and preparing a composition comprising an aggregate of gametophytes of seaweed, containing at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, wherein the weight ratio of the at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or a salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.0.

[0105] This disclosure (14) states that the above seaweed is the raw material described in this disclosure (13), which is a brown alga.

[0106] This disclosure (15) states that the above seaweed is the raw material described in this disclosure (13) or (14), which is a female gametophyte.

[0107] The present disclosure (16) is a cell tissue permeable composition comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives and linoleic acid or a salt thereof, wherein the weight ratio of the at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.0.

[0108] The present disclosure (17) is a cell tissue penetration enhancer comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives and linoleic acid or a salt thereof, wherein the weight ratio of the at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.0. [Examples]

[0109] The present invention will be described based on examples, but the present invention is not limited to these examples.

[0110] (Example 1) The method for producing the composition according to Example 1 of the present invention is described below.

[0111] (Preparation of PESI culture medium) To approximately 300 mL of distilled water, Tris, NaNO3, β-glycerophosphate disodium (Na2-glycerophosphate), Fe stock solution, P-II metal mix, and KI were added in the specified amounts in the order shown below, and thoroughly dissolved. Then the pH of the solution was adjusted to 7.8, and the volume was increased to 1000 mL with distilled water. The resulting PESI culture medium was dispensed into containers, sterilized by autoclaving (121°C, 20 minutes), and then the sterile PESI culture medium was stored refrigerated at 4°C.

[0112] The composition of the PESI culture medium is: 2-Amino-2-hydroxymethyl-1,3-propanediol(Tris(hydroxymethyl)aminomethane;Tris), 5.0g; NaNO3, 3.5g; Na2-glycerophosphate, 500 mg; Fe stock solution (see composition below), 250 mL; P-II metal mix (see composition below), 250 mL; KI stock solution (0.1mg / mL), 10mL Then, the above was diluted with distilled water to make a 1000 mL solution (pH 7.8).

[0113] The Fe stock solution consisted of 330 mg of Na2-EDTA·2H2O and 351 mg of Fe(NH4)2(SO4)2·6H2O, which were diluted with distilled water to 500 mL (Fe:EDTA molar ratio = 1:1). It was stored refrigerated at 4°C.

[0114] The composition of the P-II metal mix is: Na2-EDTA·2H2O, 500mg; H3BO3, 570mg; FeCl3·6H2O, 24.5 mg; MnSO4·4H2O, 82.0 mg; CoSO4·7H2O stock solution(4.8mg / mL), 0.5mL; ZnSO4·7H2O, 11.0 mg Then, I added all of the above ingredients to distilled water in order, and finally added distilled water to make a 500 mL solution. I stored it in the refrigerator at 4°C. Additionally, the KI stock solution was prepared by adding 200 mL of distilled water to 20 mg of KI and storing it refrigerated at 4°C.

[0115] (A. Sample preparation of female gametophytes) (1) Natural seawater was filtered through a cartridge filter with a 1 μm mesh, and the filtered seawater was heated and pressurized in an autoclave at 121°C, 2 atmospheres, and for 50 minutes to obtain 10 liters of sterile seawater. Then, 20 mL of the previously prepared PESI culture solution was added to 1 L of sterile seawater to obtain the culture seawater.

[0116] (2) Naruto wakame seaweed female gametophytes were cultured and grown from wakame collected near the mouth of the Yoshino River, following the procedure described in "(I) Cultivation of seaweed gametophytes" above. 300 mg of wakame female gametophytes and culture seawater were placed in a 5L glass Erlenmeyer flask, and preliminary aeration culture (hereinafter referred to as "preliminary culture") was performed until the wakame female gametophytes reached 10 gw.w (wet weight g). The aeration culture was carried out using the following method. First, a Pasteur pipette (IK-PAS-9P manufactured by Iwaki Glass Co., Ltd.) was attached to the tip of a silicone tube (SR-1554 manufactured by Tigers Polymer Co., Ltd.), and an air pump (APN-057R manufactured by Iwaki Corporation) was attached to the other end of the silicone tube to assemble an aeration device. Next, after immersing the tip of the Pasteur pipette in the culture seawater, aeration culture was performed by supplying air to the culture seawater from an air pump via a silicone tube and the Pasteur pipette. This aeration culture was carried out in the same manner for both the preliminary culture and the main culture. The culture conditions for the preliminary culture were a temperature of 20°C, using a daylight-white fluorescent lamp as the light source, with a light intensity of 50 μmol m³. -2 s -1 The photoperiod was set to 12L (light) and 12D (dark).

[0117] (3) 10 gw.w of pre-cultured female gametophytes of wakame and culture seawater were placed in a 20 L cylindrical polycarbonate container (manufactured by Nikko Hansen Co., Ltd.) and cultured until the volume reached 50 gw.w (main culture). The culture conditions for this culture were: temperature 20°C, green LED (wavelength 518nm) (manufactured by Nippon Medical Equipment Co., Ltd., 3LH-64) as the light source, light intensity: 50-100 μmol m -2 s -1 The photoperiod was set to 12L (light) and 12D (dark).

[0118] (4) After the main culture was completed, the female gametophytes were poured into a 50 μm mesh plankton net (Sefar Inc, DIN110) and filtered to collect the algae. The collected female gametophytes, still in the plankton net, were lightly sandwiched between commercially available paper towels to absorb and dehydrate them, and then transferred to a resealable plastic bag. The gametophytes were flattened to facilitate freeze-drying and frozen in a freezer set to -60°C. The fully frozen gametophytes were freeze-dried overnight in a freeze-dryer (Tokyo Rika Kikai Co., Ltd., FD-1) to obtain female gametophyte samples.

[0119] (B. Water extraction treatment of female gametophytes of wakame seaweed and analysis of water extracts) (B-1) Water extraction treatment of the female gametophyte of wakame seaweed The 7 mg female gametophyte sample prepared in "(A. Sample Preparation of Female Gametophyte)" above was suspended in 700 mL of distilled water and extracted on ice (0°C) for 30 minutes. Then, it was separated using a centrifuge (Hitachi-CR22GIII) at 8000 rpm for 1 hour at a temperature of 4°C, and the supernatant was collected.

[0120] (B-2) Analysis of fucoxanthin in the extract The following liquid chromatography apparatus I was prepared for the detection of fucoxanthin. Column: 3.0 mm in diameter x 150 mm in length. The packing material used is 25437-96 RP-18GP 150-3, manufactured by Kanto Chemical Co., Ltd., a high-performance liquid chromatography packing material with a particle size of 5.0 μm. Detector: Absorbance was measured using a spectrophotometer (Shimadzu Corporation UV-1800). Wavelength: 450 nm Developing solvent A: 10% acetonitrile prepared by mixing acetonitrile for LC / MS (manufactured by Kanto Chemical Co., Ltd., 01033-76) and 0.05 wt% formic acid aqueous solution in a 1:9 ratio. Developing solvent B: 80% acetonitrile prepared by mixing acetonitrile for LC / MS (manufactured by Kanto Chemical Co., Ltd., 01033-76) and 0.05 wt% formic acid aqueous solution in an 8:2 ratio. Developing solvent C: 100% acetonitrile Gradient of development time: Solvent A (0 min) → Solvent B (1 min) → Solvent C (10 min) → Solvent C (15 min) → Solvent A (15.1 min) → Solvent A (20 min) Solution flow rate: 0.5 mL / min Injection volume: 5μL Column temperature: 40℃ Sample room temperature: 15℃ Liquid transfer pump: Shimadzu Corporation LC10ADVP

[0121] Furthermore, 0.6589 mg of fucoxanthin standard (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) was weighed, and 1 L of BG was added and thoroughly mixed to prepare a 1 mM fucoxanthin solution. This 1 mL solution was then diluted 10-fold with BG to prepare a 0.1 mM (100 μM) standard solution. The retention time of fucoxanthin in the liquid chromatography apparatus I described in (1) was confirmed using this standard solution. In addition, this 100 μM fucoxanthin standard solution was diluted with BG to 2-fold (50 μM), 3.33-fold (30 μM), 10-fold (10 μM), and 33.3-fold (3 μM). Then, a calibration curve for fucoxanthin concentration in the liquid chromatography apparatus I described above was created using the standard solution and the diluted solutions.

[0122] (C. 100% butylene glycol (BG) extraction of female gametophytes of wakame after water extraction and analysis of BG extract) (C-1) BG extraction treatment of aqueous extract of female gametophyte of wakame seaweed After performing a water extraction treatment of the female gametophyte of wakame seaweed using the same procedure as in (B-1) above, the extraction residue of the female gametophyte of wakame seaweed was (1) suspended in 700 mL of 100% butylene glycol (1,3-butylene glycol, hereinafter referred to as "BG") for 7 g of solid matter accumulated at the bottom of the bottle after centrifugation, and extracted on ice (0°C) for 30 minutes. Then, it was separated using a centrifuge (Hitachi-CR22GIII) at 8000 rpm for 1 hour at 4°C, and the supernatant was collected to prepare the BG extract. Note that if extraction was performed at room temperature (25°C) for 30 minutes instead of on ice, the amount of fucoxanthin extracted would decrease slightly, and the weight ratio of fucoxanthin to linoleic acid (fucoxanthin / linoleic acid) would drop to 1.0.

[0123] (C-2) Analysis of fucoxanthin in BG extract The BG extract prepared in (C-1) above was analyzed using the liquid chromatography apparatus I described above. The liquid chromatography chart is shown in Figure 2. The horizontal axis of the chart represents time (min), and the vertical axis represents absorbance at 450 nm (dimensionless). The fucoxanthin content in the BG extract was 0.013 mg / mL. (C-3) Analysis of linoleic acid in BG extract Furthermore, linoleic acid was analyzed using an LC-MS / MS instrument (LC section: DIONEX Ultimate3000, MS / MS section: Q Exactive Focus: Thermo Fisher Scientific K.K.) under the following conditions: Column = Aclaim PR-MS 2.1 mmφ × 150 mm (Thermo Fisher Scientific K.K.), Solvent = 95% acetonitrile / aqueous acetate, Flow rate = 0.25 mL / min, Column temperature = 40°C, Detection MS (SIM), Introduction = 2 μL of sample solution. A calibration curve was created using linoleic acid (Fujifilm Wako Pure Chemical Industries, Ltd.) as a standard substance, and quantification was performed. The results are shown in Figures 3A and 3B. Figure 3A is a chart showing the results of linoleic acid analysis by LC / MS with time (min) on the horizontal axis. The peak around 4.2 minutes is the peak for linoleic acid. In Figure 3A, there appear to be four peaks from 4.17 to 4.31 minutes, but this is because a single linoleic acid sample is being measured, and a calibration curve can be created using the area of ​​the peak around 4.2 minutes. The detector used has the function of alternately measuring negatively charged molecular ions and positively charged molecular ions (alternating positive-negative scan). Linoleic acid molecular ions have a negative charge, and when negatively charged molecular ions are measured for a predetermined time, a portion of the linoleic acid peak appears on the chart. After the predetermined time has elapsed, the device then begins to measure positively charged molecular ions, causing the rise of the linoleic acid molecular ion peak to stop and creating a shoulder on the peak. When positive charges are measured for a predetermined time, the device begins to measure negatively charged molecular ions again, causing the linoleic acid peak to rise again. In this way, because positively and negatively charged molecular ions are measured alternately, a shoulder appears on the linoleic acid peak, which has a negative charge. However, what is being measured is the negatively charged linoleic acid molecular ion. Figure 3B is a chart showing the results of LC / MS analysis of linoleic acid with the mass-to-charge ratio (m / z) on the horizontal axis. The peak at m / z = 279.23 in Figure 3B is the peak of the molecular ion of linoleic acid. The linoleic acid (free linoleic acid) content in the BG extract was 0.011 mg / mL.

[0124] (D. Tape Stripping Test) (1) The fucoxanthin content in the BG extract from the female gametophyte was 0.013 mg / L, and the linoleic acid content was 0.011 mg / L, with a fucoxanthin / linoleic acid weight ratio of 1.18. Fucoxanthin (Fujifilm Wako Chemical, Fucoxanthin for Cell Biology) and / or linoleic acid (Fujifilm Wako Chemical) were appropriately added to the BG extract from the female gametophyte to prepare solutions with fucoxanthin / linoleic acid weight ratios of 1.000 (Solution 1), 9.000 (Solution 2), 19.500 (Solution 3), 28.100 (Solution 4), and 36.700 (Solution 5). (2) The skin penetration of fucoxanthin from each level of the infiltration solution was evaluated using the inner arm, from the wrist to below the elbow. First, 100 μl each of the undiluted BG extract and adjusted solutions 1-5 were dropped onto the skin surface on the inner arm and left to stand for 3 minutes to allow for penetration. (3) After 3 minutes, cover with tissue and press lightly to remove the liquid. (4) Stratum corneum release tape (CF20 Corneofix, manufactured by Integral Co., Ltd.) was prepared and pressed onto the skin surface that had come into contact with the undiluted BG extract and the adjustment solutions 1-5, and then peeled off. This was designated as the first layer. Next, a new stratum corneum release tape was applied to the same area on the skin surface and peeled off, and this was designated as the second layer. Furthermore, a new stratum corneum release tape was applied to the same area on the skin surface in the same manner and peeled off, and this was designated as the third layer. This test was performed for the undiluted BG extract and each of the adjustment solutions 1-5, and the first, second, and third layers were collected for each solution. (5) Each sheet was placed in a plastic container with 2 ml of ethanol, stirred well, and left to stand for about 30 minutes to dissolve the fucoxanthin and linoleic acid attached to the tape into the ethanol. (6) The concentrations of fucoxanthin and linoleic acid in ethanol were measured by the aforementioned liquid chromatography and LC / MS, and the weight ratio of fucoxanthin and linoleic acid was determined. The ratio of fucoxanthin in the first layer and the third layer was also calculated for each weight ratio. The results are shown in Table 1 and Figure 4.

[0125] [Table 1]

[0126] From the results in Table 1 and Figure 4 above, when the weight ratio of fucoxanthin / linoleic acid is between 1.1 and 22.0, the amount of fucoxanthin in the third layer is approximately 30% (about 1 / 3) or more of that in the first layer, indicating that linoleic acid has an effect of improving permeability. Furthermore, considering the permeability of fucoxanthin itself without linoleic acid, the weight ratio of the third layer to the first layer (= third layer / first layer) is thought to be less than 0.2, based on extrapolation of the horizontal axis in the 0 direction in Figure 4. As described above, in this invention, the permeability of fucoxanthin to the skin can be improved by adjusting the weight ratio of fucoxanthin to linoleic acid (free).

Claims

1. A composition comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, wherein the weight ratio of the at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.

0.

2. The composition according to claim 1, wherein the linoleic acid or salt thereof is derived from seaweed, comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives.

3. The composition according to claim 2, wherein the seaweed is the gametophyte of the seaweed.

4. The composition according to claim 2, wherein the seaweed is the female gametophyte of the seaweed.

5. The composition according to claim 2, wherein the seaweed is brown algae.

6. The composition according to claim 1, wherein the composition does not contain an extract of Phaeodactylum tricornutum.

7. The composition according to any one of claims 1 to 6, further comprising a polysaccharide.

8. The composition according to any one of claims 1 to 6, wherein the composition comprises an organic solvent or a mixed solvent consisting of an aqueous solvent and an organic solvent, and the organic solvent dissolves at least one compound selected from the group consisting of fucoxanthin and its derivatives.

9. The composition is a powdered composition as described in any one of claims 1 to 6.

10. A method for producing a composition comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, comprising the following steps (1) to (3). Step (1): Disperse the gametophyte of the seaweed in an aqueous solvent to obtain a dispersion of the gametophyte of the seaweed. Step (2): The aqueous solvent is separated and removed from the dispersion of the gametophyte of the seaweed obtained in step (1). Step (3): Add an organic solvent to the gametophyte of the seaweed from which the aqueous solvent has been separated and removed in step (2) to obtain a composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives and linoleic acid or a salt thereof, wherein the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to linoleic acid or a salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.

0.

11. A method for producing a composition comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, comprising the following steps (1) to (4). Step (1): Disperse the gametophyte of the seaweed in an aqueous solvent to obtain a dispersion of the gametophyte of the seaweed. Step (2): The aqueous solvent is separated and removed from the dispersion of the gametophyte of the seaweed obtained in step (1). Step (3): An organic solvent is added to the gametophyte of the seaweed from which the aqueous solvent has been separated and removed in step (2) to obtain an organic solvent composition containing at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof. Step (4): Add at least one compound selected from the group consisting of fucoxanthin and its derivatives and / or linoleic acid or a salt thereof to the organic solvent composition obtained in step (3) to obtain a composition in which the weight ratio of at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is adjusted to 1.1 to 22.

0.

12. A method for producing the composition according to claim 10 or 11, further comprising a drying step of drying the composition to make it into a powder.

13. A raw material used for extracting and preparing a composition comprising an aggregate of gametophytes of seaweed, containing at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, wherein the weight ratio of the at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or a salt thereof]) is 1.1 to 22.

0.

14. The raw material according to claim 13, wherein the seaweed is a brown algae.

15. The raw material according to claim 13, wherein the seaweed is a female gametophyte.

16. A cell tissue permeable composition comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, wherein the weight ratio of the at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or salt thereof]) is 1.1 to 22.

0.

17. A cell tissue penetration enhancer comprising at least one compound selected from the group consisting of fucoxanthin and its derivatives, and linoleic acid or a salt thereof, wherein the weight ratio of the at least one compound selected from the group consisting of fucoxanthin and its derivatives to the linoleic acid or salt thereof ([at least one compound selected from the group consisting of fucoxanthin and its derivatives] / [linoleic acid or salt thereof]) is 1.1 to 22.0.

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