Process for the production of free monounsaturated fatty acids or lower alcohol esters thereof from a water product

Abstract

A method for producing free monounsaturated fatty acids or lower alcohol esters thereof having 20 and / or 22 carbon atoms, which comprises the following steps: a step of hydrolyzing or alcoholizing an oil and fat derived from an aquatic product raw material to obtain free fatty acids or lower alcohol esters; a step of distilling the free fatty acids or lower alcohol esters to reduce the concentration of fatty acids having 18 or less carbon atoms in the free fatty acids or lower alcohol esters; and a step of fractionating a component of the free monounsaturated fatty acids or lower alcohol esters thereof having 20 and / or 22 carbon atoms by column chromatography of a reverse phase partition system.

Description

[0001] This application is a divisional application of Chinese Patent Application No. 201580026352.7, filed on July 2, 2015, entitled "Method for manufacturing free monounsaturated fatty acids or their lower alcohol esters derived from aquatic products". Technical Field

[0002] This invention relates to a method for producing free monounsaturated fatty acids (unsaturated fatty acids having one double bond, hereinafter also referred to as MUFA) or their lower alcohol esters with 20 or more carbon atoms from oils derived from aquatic products, the free monounsaturated fatty acids or their lower alcohol esters, and their uses. Background Technology

[0003] Fish oil contains unique fatty acids, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), which are polyunsaturated fatty acids (PUFAs). These fatty acids have been found to have many physiological activities and are widely used as supplements or medicines.

[0004] Vegetable oils are widely used in food. Saturated fatty acids raise blood cholesterol levels. In contrast, unsaturated fatty acids with 18 carbon atoms, namely oleic acid (mono-), linoleic acid (di-), and linolenic acid (tern-), which are abundant in vegetable oils, are of great interest because they lower cholesterol levels. In particular, oleic acid, a MUFA with 18 carbon atoms, has no effect on good cholesterol but only lowers bad cholesterol, thus it can be said to be beneficial to health. Vegetable oils also contain MUFAs with 20 and 22 carbon atoms, mostly found in rapeseed oil, violet oil, mustard oil, tung oil, etc. MUFAs with more than 20 carbon atoms are mainly called LC-MUFAs. Reported methods for manufacturing LC-MUFAs derived from vegetable oils include urea addition and recrystallization (e.g., WO89 / 08095 and Japanese Patent Application Publication No. 9-278706).

[0005] In addition, fish oil also contains monounsaturated fatty acids (hereinafter referred to as MUFAs), mainly MUFAs with 20 or 22 carbon atoms (hereinafter, MUFAs with more than 20 carbon atoms are referred to as LC-MUFAs). LC-MUFAs derived from fish oil have been reported to have physiological activities such as cholesterol-lowering effects (e.g., WO2012 / 121080). LC-MUFAs derived from vegetable oils are predominantly n-9, while those derived from fish oil are mainly n-11, indicating a difference. n-9 indicates that the 9th double bond from the methyl terminus of the fatty acid is a double bond, while n-11 indicates that the 11th double bond from the methyl terminus of the fatty acid is a double bond.

[0006] Unlike vegetable oils, oils derived from aquatic products contain, in addition to MUFAs, various fatty acids with 12 to 24 carbon atoms and 0 to 6 double bonds. As an example of refining aquatic product oils, WO2012 / 121080 describes a laboratory-standard refining method that involves ethyl esterifying saury oil and attaching it to an ODS column to concentrate MUFAs. The concentration of MUFAs with 20 and / or 22 carbon atoms derived from aquatic products obtained by this method is approximately 70%. Summary of the Invention

[0007] The problem that the invention aims to solve Besides MUFA contained in oils derived from aquatic products, substances that exhibit similar behavior to MUFA during refining processes also exist in various fatty acids with 12 to 24 carbon atoms and 0 to 6 double bonds. Therefore, it is difficult to efficiently concentrate MUFA to a high concentration, and no examples of mass production through industrially applicable methods have been found to date.

[0008] Reports indicate that LC-MUFAs, abundant in the oils of aquatic products such as saury and cod, have beneficial effects on metabolic syndrome. However, these oils also contain highly unsaturated fatty acids with more than 20 carbon atoms (hereinafter also referred to as LC-PUFAs), which poses an obstacle to correctly verifying the efficacy of LC-MUFAs or their esters. To apply LC-MUFAs or their esters, which are useful as pharmaceuticals, clinically or to a wider range of diseases, there is a need for the large-scale and efficient production of substances containing high concentrations of LC-MUFAs; or substances containing almost no components other than LC-MUFAs, for example, with an LC-MUFA concentration (purity) of 85% by weight or higher, and further, 90% by weight or higher.

[0009] In particular, LC-MUFA derived from aquatic product oils mainly consists of LC-MUFA contained in vegetable oils and isomers with different double bond positions. Since high-concentration LC-MUFA derived from aquatic product oils does not exist, a supply device is required for it.

[0010] The objective of this invention is to provide an industrial manufacturing method for efficiently obtaining high concentrations of LC-MUFA, or for efficiently obtaining LC-PUFA and LC-MUFA with low saturated fatty acid content, and the resulting high-purity LC-MUFA.

[0011] Technical solutions for solving the problem The present invention includes methods for manufacturing monounsaturated fatty acids or their lower alcohol esters with 20 and / or 22 carbon atoms, and various methods for manufacturing monounsaturated fatty acids or their lower alcohol esters with 20 and / or 22 carbon atoms.

[0012] [1] A method for producing free monounsaturated fatty acids or their lower alcohol esters with 20 and / or 22 carbon atoms, comprising the following steps: hydrolyzing or alcoholystomizing an oil derived from an aquatic product to obtain free fatty acids or lower alcohol esters; distilling the free fatty acids or lower alcohol esters to reduce the concentration of fatty acids with 18 or fewer carbon atoms in the free fatty acids or lower alcohol esters; and separating the components of free monounsaturated fatty acids or their lower alcohol esters with 20 and / or 22 carbon atoms by column chromatography of a reversed-phase partitioning system.

[0013] [2] According to the method of [1], the oil derived from aquatic raw materials is a refined oil obtained by refining crude oil obtained from aquatic products by performing at least one refining treatment selected from the group consisting of degumming, deacidification, decolorization and deodorization.

[0014] [3] The method according to [1] or [2], wherein distillation is rectification.

[0015] [4] The method according to [1] or [2], wherein the distillation is molecular distillation or short-path distillation.

[0016] [5] The method according to any one of [1] to [3], wherein the distillation is a rectification using a regular packing.

[0017] [6] The method according to any one of [1] to [5], wherein free monounsaturated fatty acids or their lower alcohol esters with a concentration of 70% by weight or more, 80% by weight or more, 90% by weight or more, or 95% by weight or more in all fatty acids are obtained by column chromatography.

[0018] [7] According to any one of [1] to [6], wherein the concentration of the free fatty acids or their lower alcohol esters with 18 or fewer carbon atoms after distillation is less than 30% by weight, less than 20% by weight, less than 10% by weight, less than 5% by weight or less than 1% by weight of all fatty acids.

[0019] [8] The method according to any one of [1] to [7], wherein the concentration of free highly unsaturated fatty acids or their lower alcohol esters after distillation and column chromatography is less than 5% by weight or less than 1% by weight of all fatty acids.

[0020] [9] A free monounsaturated fatty acid or its lower alcohol ester with 20 and / or 22 carbon atoms, which can be obtained by any one of [1] to [8].

[0021]

[10] A free fatty acid or its lower alcohol ester, comprising at least free codoleic acid (n-11) or its lower alcohol ester, or free cetearenoic acid (n-11) or its lower alcohol ester, It contains 70% or more, 80% or more or 90% or more of all fatty acids, of free monounsaturated fatty acids having 20 and / or 22 carbon atoms, or their lower alcohol esters.

[0022]

[11] According to

[10] , the free monounsaturated fatty acid or its lower alcohol ester having 20 carbon atoms is free codoleic acid (n-11) or its lower alcohol ester, and / or free stigmocarboxylic acid (n-9) or its lower alcohol ester, and the free monounsaturated fatty acid or its lower alcohol ester having 22 carbon atoms is free cetearyl acid (n-11) or its lower alcohol ester, and / or free erucic acid (n-9) or its lower alcohol ester.

[0023]

[12] A free fatty acid or its lower alcohol ester, wherein at least 70% by weight of all fatty acids are free monounsaturated fatty acids or their lower alcohol esters with 20 and / or 22 carbon atoms, and contain at least free codoleic acid or its lower alcohol ester, or free cetearyl acid or its lower alcohol ester, wherein the free saturated fatty acids or their lower alcohol esters in all fatty acids are 10% or less by weight, and the free highly unsaturated fatty acids or their lower alcohol esters in all fatty acids are 5% or less by weight.

[0024]

[13] The free fatty acid or its lower alcohol ester according to

[12] , wherein at least 90% by weight of all fatty acids are free monounsaturated fatty acids or their lower alcohol esters with 20 and / or 22 carbon atoms, and contain at least free codoleic acid or its lower alcohol ester, or free cetearyl acid or its lower alcohol ester, the free saturated fatty acids or their lower alcohol esters in all fatty acids are 5% or less by weight, and the free highly unsaturated fatty acids or their lower alcohol esters in all fatty acids are 1% or less by weight.

[0025]

[14] The free fatty acid or its lower alcohol ester according to any one of [9] to

[13] , which contains at least free codoleic acid or its lower alcohol ester, and contains 30% by weight or more, 40% by weight or more, 50% by weight or more, 60% by weight or more or 70% by weight or more of free codoleic acid (n-11) or its lower alcohol ester in all fatty acids.

[0026]

[15] The free fatty acid or its lower alcohol ester according to any one of [9] to

[14] , wherein the free fatty acid or its lower alcohol ester with 18 or fewer carbon atoms in all fatty acids is 30% or less by weight, 20% or less by weight, 10% or less by weight, 5% or less by weight, or 1% or less by weight.

[0027]

[16] The free fatty acid or its lower alcohol ester according to any one of [9] to

[15] can be obtained from the oil of aquatic products.

[0028] Use of any of the free fatty acids or their lower alcohol esters as described in any one of

[17] [9] to

[16] in food manufacturing.

[0029]

[18] A metabolic syndrome improver or lifestyle disease preventer, wherein the free fatty acid or its lower alcohol ester as described in any one of [9] to

[16] is used as the active ingredient.

[0030]

[19] A composition for improving metabolic syndrome or preventing lifestyle diseases, comprising the metabolic syndrome improver or lifestyle disease preventer described in

[18] , and additives.

[0031] Use of any one of the free fatty acids or their lower alcohol esters described in

[20] , [9] to

[16] in the manufacture of the metabolic syndrome improvement composition or lifestyle disease prevention composition described in

[18] .

[0032] Invention Effects According to the method of the present invention, high concentrations of LC-MUFA can be obtained efficiently. Furthermore, according to one aspect of the present invention, LC-PUFA derived from aquatic product oils can be provided with low saturated fatty acid content, high concentration, and high yield. According to one aspect of the present invention, the content of saturated fatty acids and LC-PUFA in LC-MUFA compositions can be reduced. When using LC-MUFA as a functional ingredient, this method is suitable for applications seeking low concentrations of saturated fatty acids and / or LC-PUFA. Fatty acids exhibit significant differences not only in their physical properties but also in their physiological functions when the number of carbon atoms or double bonds differs. Since LC-MUFA derived from aquatic product oils, which is currently difficult to separate, can be separated according to the number of carbon atoms, their respective functions can be clearly defined and effectively utilized. This method is suitable for use in pharmaceuticals, supplements, etc., where LC-MUFA is an active ingredient. Detailed Implementation

[0033] In this specification, "oil" or "grease" refers not only to triglycerides, but also to crude oils that are mainly composed of triglycerides and contain other lipids such as diglycerides, monoglycerides, phospholipids, cholesterol, and free fatty acids. "Oil" or "grease" refers to compositions containing these lipids.

[0034] The term "fatty acid" includes not only free saturated or unsaturated fatty acids themselves, but also the fatty acids that serve as structural units contained in free saturated or unsaturated fatty acids, saturated or unsaturated fatty acid alcohol esters, triglycerides, diglycerides, monoglycerides, phospholipids, sterol esters, etc. It can also be referred to as structural fatty acids. In this specification, unless otherwise specified, the manner in which a compound contains fatty acids is sometimes omitted. Examples of ways in which a compound contains fatty acids include free fatty acid form, fatty acid alcohol ester form, glyceride form, phospholipid form, and sterol ester form. A compound containing the same fatty acid may be present in oil in a single manner or as a mixture of two or more manners.

[0035] Experience has shown that the hydrolysis or alcoholysis of fatty acids is highly efficient, primarily yielding compositions of fatty acids in the form of free fatty acids or their lower alcohol esters. Therefore, unless otherwise specified, descriptions of the composition and whether the fatty acids are in the form of free fatty acids or lower alcohol esters are sometimes omitted when discussing the fatty acids after processing. However, the presence of fatty acids in forms other than free fatty acids or lower alcohol esters is not entirely excluded.

[0036] When describing fatty acids, the following numerical representation is sometimes used: simplified representations of the number of carbon atoms, the number of double bonds, and the location of the double bonds are expressed using both numbers and letters. For example, a saturated fatty acid with 20 carbon atoms can be expressed as "C20:0", a monounsaturated fatty acid with 18 carbon atoms can be expressed as "C18:1", and arachidonic acid can be expressed as "C20:4, n-6". "n-" indicates the position of the double bond from the methyl terminus of the fatty acid; for example, "n-6" indicates that the double bond is the 6th bond from the methyl terminus. This method is well known to those skilled in the art, and any fatty acid described according to this method can be readily defined by such a person.

[0037] In this specification, "crude oil" refers to the mixture of the above-mentioned lipids, i.e., oil in a biologically extracted state. In this specification, "refined oil" refers to oil obtained by refining crude oil through a refining process that removes substances other than target substances such as phospholipids and sterols, by subjecting the crude oil to at least one of the following processes: degumming, deacidification, decolorization, and deodorization.

[0038] In this invention, "oils obtained from aquatic products" or "oils derived from aquatic raw materials" may include lipids such as oils, phospholipids, and wax esters found in fish, crustaceans, or marine animals. Examples of fish rich in LC-MUFA include: saury and other fish belonging to the Pacific saury family; cod, pollock, Atlantic cod, sable scad and other fish belonging to the cod family; salmon, coho salmon, sockeye salmon, lenok salmon, Atlantic salmon, rainbow trout and other salmonids; capelin, gudgeon and other smeltids; herring and other herring. In addition, needlefish, tuna, mackerel, red snapper, bulleye snapper, pinefish, Pacific bass, and blue scorpionfish also contain significant amounts. Furthermore, the liver oil of sharks such as North Pacific dogfish, basking shark, and blackfin shark also contains a considerable amount. Oils derived from animals such as seals and whales can also be utilized. Even with limited raw materials, LC-MUFA can be concentrated and used.

[0039] The fifth revision of the Japanese Food Standards Composition Tables lists that the fatty acids in raw saury contain 19.3% by weight of docosanoic acid (C22:1), 17.2% by weight of eicosanoic acid (C20:1), and a total of 50.1% by weight of monounsaturated fatty acids. A characteristic of saury oil is that, even in fish oil, it has a high content of monounsaturated fatty acids. Among the fatty acids, fish oil containing at least 10% by weight of docosanoic acid and at least 15% by weight of eicosanoic acid are preferred. Fish oil from fish with high catch volumes, such as saury and cod, is preferred as the raw material.

[0040] In this invention, "monounsaturated fatty acid" or "MUFA" refers to a fatty acid with one double bond, and "highly unsaturated fatty acid" or "PUFA" refers to a fatty acid with four or more double bonds. MUFA or PUFA, i.e., long-chain fatty acids with 20 or more carbon atoms, are denoted as LC-MUFA or LC-PUFA.

[0041] In LC-MUFA, in this invention, MUFAs with 20 and / or 22 carbon atoms, especially n-11 isomers, can be obtained at high concentrations.

[0042] MUFAs with 20 carbon atoms are called icosene acids under the name IUPAC, and are further classified according to the position of the double bond as cis-icos-9-enoic acid (n-11, commonly known as gadoleic acid) and cis-icos-11-enoic acid (n-9, commonly known as gondoic acid). MUFAs with 22 carbon atoms are called dodecenoic acids under the name IUPAC, and are further classified according to the position of the double bond as cis-docos-11-enoic acid (n-11, commonly known as ceteneic acid) and cis-docos-13-enoic acid (n-9, commonly known as erucic acid). Aquatic oils contain large amounts of n-11 gadoleic acid and / or ceteneic acid.

[0043] Besides MUFAs, other fatty acids found in fish oil and similar products include saturated fatty acids (14, 16, 18, 20 carbon atoms, etc.), binary and tricyclic unsaturated fatty acids (18, 20 carbon atoms, etc.), and highly unsaturated fatty acids with four or more carbon atoms (PUFAs, 20, 22 carbon atoms, etc.). Among these, highly unsaturated fatty acids are characteristic fatty acids found in fish oil and similar products, such as fatty acids with more than 20 carbon atoms and more than 4 double bonds. Specifically, examples include arachidonic acid (20:4, n-6), eicosapentaenoic acid (20:5, n-3), docosapentaenoic acid (22:5, n-6), and docosahexaenoic acid (22:6, n-3).

[0044] In this invention, the ester of MUFA refers to a lower alcohol of MUFA with 1 to 3 carbon atoms, preferably an ester of MUFA ethanol.

[0045] One aspect of the present invention provides a method for producing free monounsaturated fatty acids or their lower alcohol esters with 20 and / or 22 carbon atoms, comprising: hydrolyzing or alcoholystolytically digesting oils derived from aquatic raw materials to obtain free fatty acids or lower alcohol esters (hereinafter, sometimes referred to as a processing step); distilling the free fatty acids or lower alcohol esters to reduce the concentration of fatty acids with 18 or fewer carbon atoms in the free fatty acids or lower alcohol esters (hereinafter, sometimes referred to as a distillation step); and separating the components of free monounsaturated fatty acids or their lower alcohol esters with 20 and / or 22 carbon atoms by column chromatography of a reversed-phase partitioning system (hereinafter, sometimes referred to as a column step), and may include other steps as appropriate.

[0046] The method for obtaining crude oil from various aquatic raw materials can be any method. Taking Pacific saury crude oil as an example, it is usually obtained in the same way as other fish oils, using the following method: After crushing and steaming the whole Pacific saury or processing residues such as the head, skin, backbone, and viscera from aquatic product processing, it is pressed to separate the cooking liquid (mucus) and the pressed crude oil. The oil obtained together with the cooking liquid is separated from the cooking liquid by centrifugation to produce Pacific saury crude oil.

[0047] Typically, crude fish oil is refined into refined fish oil through refining processes such as degumming, deacidification, decolorization using activated clay or activated carbon, water washing, and deodorization based on steam distillation, depending on the raw material. This refined fish oil can be used as a raw material in this invention. In other words, the oil derived from aquatic raw materials used in one aspect of the method of this invention can be a refined oil obtained by performing such general refining processes on crude oil derived from aquatic products. For example, refined oil obtained by refining crude oil derived from aquatic products through at least one of the refining processes of degumming, deacidification, and decolorization can be used as an oil derived from aquatic raw materials.

[0048] One aspect of the present invention involves a processing step of decomposing oils derived from aquatic products into free fatty acids or lower alcohol esters via hydrolysis or alcoholysis. Hydrolysis involves adding water and a catalyst or enzyme, such as an acid, to the oil and reacting to release fatty acids bonded to glycerol. Alcoholysis involves adding a lower alcohol (preferably ethanol) with 1 to 3 carbon atoms, and a catalyst or enzyme to the oil and reacting to generate fatty acids bonded to glycerol and esters of the lower alcohol. By separating the free fatty acids or lower alcohol esters from glycerol, the desired fatty acids can be concentrated.

[0049] The distillation step of one aspect of the present invention is a process of reducing the concentration of fatty acids with 18 or fewer carbon atoms in the free fatty acids or lower alcohol esters produced in the processing step by distillation. Here, by removing fatty acids with 18 or fewer carbon atoms as much as possible, the subsequent column process can function more effectively. The fatty acids with 18 or fewer carbon atoms are preferably reduced to less than 30 area%, less than 20 area%, less than 10 area%, less than 5 area%, or less than 1 area%, in other words, less than 30 wt%, less than 20 wt%, less than 10 wt%, less than 5 wt%, or less than 1 wt%. As shown in the results in Tables 2 and 3 of the examples, if the concentration of fatty acids with 18 or fewer carbon atoms is further reduced by distillation such as rectification, the C20:1 component can be further concentrated in the column process.

[0050] The distillation method can be any method, among which, a method that can remove fatty acids with 18 or fewer carbon atoms as much as possible is preferred. Examples of such distillation methods include molecular distillation, short-path distillation, and other single distillation; and rectification, with rectification being particularly preferred. Both rectification and single distillation are preferably thin-film distillation with a thin-film heated evaporator.

[0051] To separate LC-MUFAs from fatty acids with 18 or fewer carbon atoms, rectification offers better separation than single distillation methods such as short-path distillation and molecular distillation, thus efficiently reducing fatty acids with fewer than 18 carbon atoms. Rectification conditions can be adjusted by considering the fatty acid composition of the lower alcohol esters used as feedstock. Preferred rectification conditions include a bottom temperature of 220°C or lower, preferably 150–220°C, and particularly preferably 150–200°C. Reduced pressure is 10 mmHg or lower, more preferably 1 mmHg or lower, and even more preferably 0.1 mmHg or lower. The lower limit of pressure is not particularly limited and can be appropriately set depending on the apparatus used. The internal structure used to increase the theoretical number of separation stages can be of various types, such as packed or tray-type. A packed structure using a regular packing material is more preferred. Single distillation methods such as short-path distillation and molecular distillation offer superior productivity compared to rectification and are suitable for large-scale processing. Preferred distillation conditions, especially for short-path distillation or molecular distillation, include an evaporation surface temperature of 120°C or lower, preferably 50–120°C, and more preferably 50–80°C. Preferred reduced pressure conditions for single distillation, especially short-path distillation or molecular distillation, are 0.05 mmHg or less, more preferably 0.0013 mmHg or less. There is no particular limitation on the lower limit of the pressure; it can be set appropriately depending on the apparatus used.

[0052] The column step of one aspect of the present invention is a step for concentrating MUFAs with 20 and / or 22 carbon atoms, or separating them from other unsaturated fatty acids. For separating MUFAs with 20 and / or 22 carbon atoms from other unsaturated fatty acids, a reversed-phase partitioning column chromatography method is used. Specifically, an ODS column is preferred. As the stationary phase, any adsorbent in a reversed-phase partitioning system can be used without special specification, but an ODS column containing octadecylsilyl (ODS) is preferred. The adsorbent dosage is preferably 10 times or more by weight of the amount of feedstock supplied for column chromatography, more preferably 100 times or more by weight. There is no particular upper limit to the adsorbent dosage; for example, it can be set to 1000 times the amount. Regarding the eluent and mobile phase, various polar solvents such as methanol, ethanol, 2-propanol, acetone, and acetonitrile, or aqueous solvents containing water added to these polar solvents, are used; methanol is preferred.

[0053] By sequentially performing column processing steps starting from the above-mentioned processing steps, in other words, by performing distillation and chromatography, free MUFAs or their lower alcohol esters with 20 and / or 22 carbon atoms in all fatty acids can be concentrated to a high concentration. For example, they can be concentrated individually or in combination to a total of 70% by weight or more, 80% by weight or more, 90% by weight or more, or 95% by weight or more, and in all fatty acids, they can be less than 99.99% by weight or less than 99.9999% by weight.

[0054] By performing distillation and chromatography, the concentration of free fatty acids or their lower alcohol esters with 18 or fewer carbon atoms in all fatty acids can be reduced, for example, individually or in combination, to less than 30% by weight, less than 20% by weight, less than 10% by weight, less than 5% by weight, or less than 1% by weight.

[0055] By implementing distillation and chromatography, the proportion of free saturated fatty acids or their lower alcohol esters in all fatty acids can be reduced to less than 10% by weight, less than 5% by weight, or less than 1% by weight.

[0056] By implementing distillation and chromatography, the proportion of PUFA in all fatty acids can be reduced to less than 5% by weight or less than 1% by weight.

[0057] For the obtained free fatty acids or their lower alcohol esters, in order to obtain them from the oils and fats of aquatic products, the highly concentrated 20-carbon free monounsaturated fatty acids or their lower alcohol esters may be free codoleic acid (n-11) and / or free teratosuccinic acid (n-9) or their lower alcohol esters, and the highly concentrated 22-carbon free monounsaturated fatty acids or their lower alcohol esters may be free ceteneic acid (n-11) or their lower alcohol esters and / or free erucic acid (n-9) or their lower alcohol esters. The obtained free fatty acids or their lower alcohol esters may contain at least free codoleic acid (n-11) or their lower alcohol esters, or free ceteneic acid (n-11) or their lower alcohol esters, and may contain at least free codoleic acid or their lower alcohol esters.

[0058] Specifically, regarding free fatty acids or their lower alcohol esters, the total fatty acids may contain more than 30% by weight, more than 40% by weight, more than 50% by weight, more than 60% by weight, or more than 70% by weight of free codoleic acid or its lower alcohol esters, and the total fatty acids may contain less than 99.99% by weight or less than 99.9999% by weight of free codoleic acid or its lower alcohol esters.

[0059] With regard to free fatty acids or their lower alcohol esters, the total fatty acids may contain more than 30% by weight, more than 40% by weight, more than 50% by weight, more than 60% by weight, or more than 70% by weight of free cetene or its lower alcohol esters, and the total fatty acids may contain less than 99.99% by weight or less than 99.9999% by weight of free cetene or its lower alcohol esters.

[0060] With regard to free fatty acids or their lower alcohol esters, the total fatty acids may contain more than 5% by weight, more than 10% by weight, more than 15% by weight, or more than 20% by weight of free saturated cetyl acid or its lower alcohol esters, and the total fatty acids may contain less than 99.99% by weight or less than 99.9999% by weight of free saturated cetyl acid or its lower alcohol esters.

[0061] With regard to free fatty acids or their lower alcohol esters, the total fatty acids may contain more than 1%, more than 2%, or more than 3% by weight of free erucic acid or its lower alcohol esters, and the total fatty acids may contain less than 99.99% by weight or less than 99.9999% by weight of free erucic acid or its lower alcohol esters.

[0062] When free codoleic acid and free cetylene acid or their lower alcohol esters coexist in free fatty acids or their lower alcohol esters, their total concentration in all fatty acids may be set to less than 99.99% by weight or less than 99.9999% by weight.

[0063] In the presence of two or more free codoleic acid, free cetearic acid, free squalinoic acid, and free erucic acid, or these lower alcohol esters, in free fatty acids or their lower alcohol esters, their total concentration in all fatty acids may be set to less than 99.99% by weight or less than 99.9999% by weight.

[0064] Such free fatty acids or their lower alcohol esters are preferred as pharmaceuticals or supplements with LC-MUFA as the active ingredient.

[0065] Free fatty acids or their lower alcohol esters containing at least free codoleic acid or its lower alcohol esters, or free cetearenoic acid or its lower alcohol esters, and at least 70% by weight of all fatty acids being MUFAs with 20 and / or 22 carbon atoms, which can be manufactured by one method of the present invention, are preferably used as pharmaceuticals or supplements with LC-MUFAs as the active ingredient.

[0066] The following, in one aspect of the present invention, are preferred pharmaceuticals or supplements containing at least free codoleic acid or its lower alcohol esters, or free cetearyl acid or its lower alcohol esters, wherein at least 70% by weight of all fatty acids are MUFAs having 20 and / or 22 carbon atoms, and saturated fatty acids are 10% or less and PUFAs are 5% or less; or, more preferably, free fatty acids or their lower alcohol esters containing at least free codoleic acid or its lower alcohol esters, or free cetearyl acid or its lower alcohol esters, wherein at least 90% by weight of all fatty acids are MUFAs having 20 and / or 22 carbon atoms, and saturated fatty acids are 5% or less and PUFAs are 1% or less.

[0067] In particular, for use as a pharmaceutical product, it is preferred to contain at least free codoleic acid or its lower alcohol ester, or free cetearyl acid or its lower alcohol ester, at least 90% by weight of all fatty acids being MUFAs with 20 and / or 22 carbon atoms, less than 1% by weight of saturated fatty acids, less than 1% by weight of PUFAs, and free fatty acids or their lower alcohol esters.

[0068] Of these free fatty acids or their lower alcohol esters, free fatty acids or their lower alcohol esters comprising 18 or fewer carbon atoms in all fatty acids may be expressed individually or in combination as less than 30% by weight, less than 20% by weight, less than 10% by weight, less than 5% by weight, or less than 1% by weight.

[0069] These high-purity compositions of one aspect of the present invention are indispensable for the research, development, and commercialization of isomers obtained solely from aquatic products such as codoleic acid (n-11) and cetearyl acid (n-11).

[0070] The free fatty acid or lower alcohol ester of one embodiment of the present invention contains at least codoleic acid or ceteneic acid, and contains a high concentration of free monounsaturated fatty acids having 20 and / or 22 carbon atoms. Therefore, it is preferably suitable for the research, development, and commercialization of substances derived from aquatic products, such as codoleic acid or ceteneic acid. Based on the above, the free fatty acid or lower alcohol ester of one embodiment of the present invention is more preferably used when the concentration of at least one fatty acid selected from the group consisting of fatty acids with 18 or fewer carbon atoms, saturated fatty acids, and highly unsaturated fatty acids is low.

[0071] In one aspect of this invention, the free fatty acids can also be their salts. Examples of salts include potassium salts and sodium salts.

[0072] As described above, the composition of one aspect of the present invention contains a high concentration of LC-MUFA, and depending on the circumstances, the content of saturated fatty acids, LC-PUFA, etc., can be made extremely low. Therefore, it is extremely useful for applications requiring high concentrations of LC-MUFA. Examples of such applications include food, supplements, and pharmaceuticals. It is particularly preferred for applications aimed at the functionality of LC-MUFA, such as the improvement of metabolic syndrome and the prevention of lifestyle diseases.

[0073] In other aspects of the present invention, a method for improving metabolic syndrome is provided, comprising: administering a composition of one aspect of the present invention as a metabolic syndrome improver to a subject seeking to improve metabolic syndrome in an amount effective in improving metabolic syndrome.

[0074] Other aspects of the present invention include a method for preventing lifestyle diseases, comprising: administering a composition of one aspect of the present invention as a lifestyle preventive agent to a subject seeking to prevent lifestyle diseases in an amount effective for lifestyle prevention.

[0075] Humans and animals can be examples of subjects for drug administration.

[0076] For example, the compositions of one aspect of the present invention as described in (a) to (c) below can be used as metabolic syndrome modifiers or lifestyle disease preventatives in these methods of improvement or prevention: (a) A composition containing free fatty acids or their lower alcohol esters, wherein the free fatty acids or their lower alcohol esters contain at least free codoleic acid (n-11) or its lower alcohol ester, or free cetearenoic acid (n-11) or its lower alcohol ester, and contain monounsaturated fatty acids or their lower alcohol esters with 20 and / or 22 carbon atoms in an amount of 70% by weight or more, 80% by weight or more or 90% by weight of all fatty acids; (b) A composition containing free fatty acids or their lower alcohol esters, wherein at least 70% by weight of all fatty acids are monounsaturated fatty acids having 20 and / or 22 carbon atoms, and at least contains free codoleic acid or its lower alcohol ester, or free cetearic acid or its lower alcohol ester, wherein saturated fatty acids comprise less than 10% by weight of all fatty acids, and highly unsaturated fatty acids comprise less than 5% by weight of all fatty acids; and (c) A composition containing free fatty acids or their lower alcohol esters, wherein at least 90% by weight of all fatty acids are monounsaturated fatty acids having 20 and / or 22 carbon atoms, and at least contains free codoleic acid or its lower alcohol ester, or free cetearyl acid or its lower alcohol ester, wherein saturated fatty acids comprise less than 5% by weight of all fatty acids, and highly unsaturated fatty acids comprise less than 1% by weight of all fatty acids.

[0077] Refined oils derived from fish contain approximately 30% by weight of monounsaturated fatty acids with 20 and / or 22 carbon atoms or their lower alcohol esters. These refined oils derived from fish have a therapeutic effect on metabolic syndrome (e.g., see WO2012 / 121080; Lipids (2011) Vol.46, pp.425-434; J. Agric. Food Chem., 2011, Vol.59, pp.7482-7489; Lipids in Health and Disease, 2011, vol.10, pp.189-199, etc.). The metabolic syndrome improver or lifestyle disease preventer of one aspect of the present invention contains at least free codoleic acid or its lower alcohol ester, or free cetearyl acid or its lower alcohol ester, and contains a higher concentration, for example, more than 70% by weight of free monounsaturated fatty acids with 20 and / or 22 carbon atoms or their lower alcohol esters. Therefore, a higher metabolic syndrome improver or lifestyle disease preventer effect can be expected.

[0078] According to other aspects of the present invention, compositions for improving metabolic syndrome or preventing lifestyle diseases containing the metabolic syndrome improvers or lifestyle disease preventers described in (a) to (c) above, and additives, can be provided. Examples of additives include, for instance, medically permissible bases, carriers, excipients, disintegrants, lubricants, and colorants, when used as pharmaceutical products. The metabolic syndrome improvers or lifestyle disease preventers of other aspects of the present invention are preferably provided as tablets or capsules after being processed into soft capsules such as gelatin or powdered oils. The metabolic syndrome improvers or lifestyle disease preventers can be manufactured by additional steps such as combining the metabolic syndrome improvers or lifestyle disease preventers described in (a) to (c) above, and additives in a prescribed ratio, and processing them into a desired dosage form as needed. The content of metabolic syndrome improver or lifestyle disease preventer in the composition for improving metabolic syndrome or the composition for preventing lifestyle diseases is not particularly limited as long as it is a content that can be expected to have an effect. For example, it can be set to 0.01% to 100% by weight, 0.1% to 100% by weight, or 3% to 100% by weight.

[0079] The metabolic syndrome improvers or lifestyle disease preventers described in (a) to (c) above can be used as food ingredients in food. Food refers to all foods including beverages, including general foods such as supplements and other health foods, as well as specific health foods or nutritional functional foods as defined by the Consumers Association's Health Functional Food Regulations. For example, it can be a functional food displaying a message about its metabolic syndrome-improving effect or its lifestyle disease-preventing effect. When the metabolic syndrome improvers or lifestyle disease preventers described in (a) to (c) above are used as food ingredients, the food containing them can be manufactured through additional processes such as combining them with other food ingredients as needed and shaping them into any form. There are no particular restrictions on the content of metabolic syndrome improvers or lifestyle disease preventers in food; the content can be set to the desired effect. In addition to food, they can also be used as additives in animal feed as food ingredients.

[0080] When administering the drug as a medicine or food to a recipient, the dosage should be appropriately set according to the severity of symptoms, the recipient's age, weight, and health status. For example, for adults, the composition can be administered orally or non-orally once a day, or two to four times a day, or more than four times a day, at appropriate intervals, at an amount of 1 mg to 1 g / kg / day, preferably 5 mg to 300 mg / kg / day, as the effective ingredient.

[0081] In this specification, the term "process" is not limited to independent processes. Even if it cannot be clearly distinguished from other processes, it is included in this terminology as long as it can achieve the desired purpose of the process.

[0082] In this specification, the numerical range represented by “~” indicates the range in which the values ​​before and after it are respectively the minimum and maximum values.

[0083] In this specification, the amount of each component in the composition refers to the total amount of the multiple substances present in the composition, unless otherwise specified. In this specification, the terms "below" or "less than" regarding percentages, unless otherwise specified, refer to a range including 0% or values ​​that cannot be detected by current methods.

[0084] In this specification, the presence of one or more elements marked with the indefinite article "a" or "an" cannot be ruled out unless explicitly shown or combined in the context. Therefore, the indefinite article "a" or "an" usually refers to "at least one".

[0085] The verb "comprising" and its application as described in this specification are used in a non-limiting sense and include the matter following the term, excluding matters not specifically mentioned.

[0086] In this specification, the inventive features described in one embodiment of each aspect of the invention can be arbitrarily combined to form new embodiments, which should be understood as being included in the various aspects of the invention.

[0087] The present invention will now be described in detail through examples. However, the present invention is not limited thereto. Furthermore, in the following examples, unless otherwise specified, "%" refers to "weight %".

[0088] Example Fatty acid composition can be determined by conventional methods. Specifically, the oils and fats to be measured are esterified using lower alcohols and catalysts to obtain lower alcohol esters of fatty acids. Then, the obtained lower alcohol esters of fatty acids are analyzed by gas chromatography. In the obtained gas chromatogram, the peaks corresponding to each fatty acid are identified, and the peak area of ​​each fatty acid is calculated using the integration algorithm (modified C.01.03

[37] , Agilent Technologies) of Agilent ChemStation. Peak area refers to the ratio (area%) of the peak area of ​​each component to the total peak area in the chart obtained by analyzing oils and fats with various fatty acids as components using gas chromatography, thin layer chromatography / flame ionization detector (TLC / FID), etc., which indicates the content ratio of the peak component. The value of area% obtained by the above determination method is the same as the value of weight% of each fatty acid in the sample and can be used interchangeably. Referencing the Japanese Oil Chemists' Society (JOCS) Standard Oil and Fat Analysis Test Method 2013, Version 2.4.2.1-2013, Fatty Acid Composition (FID Isothermal Gas Chromatography), and the Japanese Oil Chemists' Society (JOCS) Standard Oil and Fat Analysis Test Method 2013, Version 2.4.2.2-2013, Fatty Acid Composition (FID Heated Gas Chromatography).

[0089] Fatty acid composition was analyzed by gas chromatography using the methods shown in the examples, and lipid composition was analyzed using TLC / FID. Detailed conditions are shown in the examples.

[0090] Empirically, the alkyl esterification method used in the examples has been found to achieve an ethyl esterification rate of 95% to 100%. Therefore, in the examples obtained, the ethyl esters are presumed to contain almost entirely saturated or unsaturated fatty acids in the form of fatty acid ethyl esters. Therefore, hereafter, all saturated or unsaturated fatty acids contained in the samples will be described as saturated or unsaturated fatty acids in the form of ethyl esters. However, the presence of fatty acids in forms other than free fatty acids or lower alcohol esters is not completely excluded.

[0091] [Example 1] 4000 kg of crude saury oil obtained from fresh saury was degummed, deacidified, and decolorized to obtain 3520 kg of refined saury oil. 2000 kg of the obtained refined saury oil was subjected to ester exchange with sodium ethoxide to achieve ethyl esterification, and 0.5% vitamin E was added as an antioxidant to obtain 1999 kg of ethyl saury oil (sample A). Table 1 shows the analytical values ​​of the obtained ethyl saury oil.

[0092] [Table 1] [Example 2] Refining of Saury Oil Ethyl Ester Based on Distillation 100.06 g of saury oil ethyl ester was added to a 500 mL three-necked flask. A vacuum-jacketed distillation column (with vacuum-jacketed fractionating tubes (Kiriya Works) and fractionating head (Kiriya Works)) manufactured by Kiriyama Corporation, fitted with five experimental EX (25 mm × 50 mm, Sulzer Chemtech) inserts, was used for precision distillation. The conditions were: bottom temperature 185 °C, bottom pressure 0.8 mmHg (approximately 107 Pa), top pressure 8 Pa, and top vapor temperature 133 °C. This yielded 43.2 g of purified distillate (sample B) and 54.5 g of purified residue (sample C). Table 2 shows the fatty acid composition of samples B and C.

[0093] [Example 3] Refining of Pacific saury oil ethyl esters based on molecular distillation 905.5 g of saury ethyl oil was added to a centrifugal molecular distillation apparatus (MS-150) manufactured by Nippon Shokurik Co., Ltd., and distilled at an evaporation surface temperature of 90 °C and a pressure of 0.015 Torr, yielding 209.7 g of distillate and 596.2 g of distillation residue (sample D). Table 2 shows the fatty acid composition of sample D.

[0094] [Table 2] [Example 4] Purification of MUFA by HPLC after distillation Samples C and D, containing fatty acid ethyl esters purified by distillation or molecular distillation, were further purified by ODS (OctaDecyl Silyl)-HPLC. It should be noted that the separation conditions are as follows.

[0095] Separation conditions Column: JAIGEL-ODS-AP-30, SP-120-15 (Japan Analysis Industries Co., Ltd.), 30φ×200mm Eluent: Methanol Flow rate: 20 mL / min Column temperature: 40℃ Sampling load: 6.30g Detector: Differential refractometer Using the peak value and holding time of the differential refractometer as indicators, components containing C20:1 (hereinafter referred to as C20:1 components) and components containing C22:1 (hereinafter referred to as C22:1 components) were separated from samples E to H below. Table 3 shows the fatty acid composition of each component. The fatty acid composition (%) is as described above, and is the area ratio of the chart based on gas chromatography.

[0096] Sample E: A sample of component C20:1 was obtained using sample C as the raw material. Sample F: A sample of component C22:1 was obtained using sample C as the raw material. Sample G: A sample of component C20:1 was obtained using sample D as the raw material. Sample H: A sample of component C22:1 obtained from sample D as raw material. [Table 3] As shown in Table 3, the content ratio of LC-MUFA can be increased for samples E to H by performing ODS-HPLC purification. In particular, for samples E, F, and G, the concentration can be concentrated to over 95% by weight.

[0097] On the other hand, when using sample C, which is distilled as the initial purification step, as raw material, compared to using sample D, which is distilled as raw material, the LC-MUFA with 20 carbon atoms can be further concentrated after the purification step (samples E and F). This is because, in the distillation step, there is less residue of fatty acids with 18 or fewer carbon atoms, and in the HPLC step, the reduction in the separation efficiency of C20:1 caused by the presence of fatty acids with 18 or fewer carbon atoms is effectively suppressed. It can be seen that increasing the concentration of C20:1 by selecting the initial distillation step is effective. Therefore, it can be seen that in order to obtain LC-MUFA ethyl ester with a high concentration of C20:1, removing fatty acids with 18 or fewer carbon atoms and saturated fatty acids in the purification step before HPLC purification is effective.

[0098] Even more surprisingly, it was found that by removing fatty acids with fewer than 18 carbon atoms and saturated fatty acids through distillation, LC-MUFA reached a higher concentration. When compared with sample E and sample G, which had the same concentration of LC-MUFA, the recovery rate of LC-MUFA was 1.34 times.

[0099] As shown in this embodiment, the composition obtained by the method of the present invention contains LC-MUFA at a high concentration and has extremely low contents of saturated fatty acids and LC-PUFA. Therefore, it is extremely useful for applications requiring high concentrations of LC-MUFA. Examples of such applications include food, supplements, and pharmaceuticals. It is particularly preferred for applications where the functionality of LC-MUFA is desired, such as for improving metabolic syndrome and preventing lifestyle diseases.

[0100] The entire disclosure of Japanese Patent Application No. 2014-136436, filed on July 2, 2014, is incorporated herein by reference.

[0101] All documents, patent applications, and technical specifications described in this specification are incorporated herein by reference, as are the specific and individual descriptions included in each document, patent application, and technical specification.

Claims

1. A method for producing free monounsaturated fatty acids or their lower alcohol esters with 20 and / or 22 carbon atoms, comprising the following steps: The process of hydrolyzing or alcoholyzing oils derived from aquatic raw materials to obtain free fatty acids or lower alcohol esters. The process of distilling the free fatty acid or lower alcohol ester to reduce the concentration of fatty acids with 18 or fewer carbon atoms in the free fatty acid or lower alcohol ester. as well as The process of separating free monounsaturated fatty acids or their lower alcohol esters with a concentration of 90% by weight or more of carbon atoms from all fatty acids by column chromatography using a reversed-phase partitioning system.

2. The method according to claim 1, wherein, Oils derived from aquatic raw materials are refined oils obtained by refining crude oils obtained from aquatic products, wherein the refining process is selected from at least one of the group consisting of degumming, deacidification, decolorization and deodorization.

3. The method according to claim 1, wherein, Distillation is a process of rectification.

4. The method according to claim 1, wherein, Distillation is either molecular distillation or short-path distillation.

5. The method according to any one of claims 1 to 3, wherein, Distillation is the rectification process using a regular packing material.

6. The method according to any one of claims 1 to 4, wherein, By column chromatography, free monounsaturated fatty acids with 20 and / or 22 carbon atoms, or their lower alcohol esters, with a concentration of more than 95% by weight, are obtained from all fatty acids.

7. The method according to any one of claims 1 to 4, wherein, The concentration of free fatty acids with 18 or fewer carbon atoms or their lower alcohol esters after distillation is less than 5% by weight of all fatty acids.

8. The method according to any one of claims 1 to 4, wherein, The concentration of free highly unsaturated fatty acids or their lower alcohol esters after distillation and column chromatography is less than 1% by weight of all fatty acids.

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