Method for improving stability of anserine, use of quercetin or glucoside thereof for improving stability of anserine, and composition improved in stability of anserine

Abstract

To provide a method for inhibiting temporal depletion (decomposition) of anserine due to heat and improving stability of anserine, or the like.SOLUTION: A method for improving stability of anserine includes a step of mixing anserine and quercetin or glucoside thereof such that a weight ration of quercetin or glucoside thereof on an isoquercitrin basis relative to anserine in the composition (quercetin or glucoside thereof on an isoquercitrin basis / anserine) becomes 5 or higher.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a method for improving the stability of anserine, the use of quercetin or a glycoside thereof for improving the stability of anserine, and a composition having improved stability of anserine. [Background technology]

[0002] Imidazole peptides such as anserine are widely distributed in the human body, and are particularly abundant in the muscles of vertebrates. Therefore, they are often used in oral compositions such as health foods intended to suppress muscle weakness. For example, Patent Document 1 discloses an oral composition containing an imidazole peptide such as anserine, which is an oral liquid composition containing a high concentration of collagen, and which reduces the odor characteristic of collagen by incorporating an imidazole peptide such as anserine. Patent Document 2 also describes a tablet containing glucosamines and amino acids, which further incorporates anserine to achieve good shape stability. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Japanese Patent Publication No. 2023-156812 [Patent Document 2] Japanese Patent Application Laid-Open No. 2018-35143 Summary of the Invention [Problem to be solved by the invention]

[0004] However, anserine decomposes due to heat and humidity, decreasing in content over time. Therefore, when anserine-containing compositions (particularly oral compositions such as health foods) are stored for a long period of time under inappropriate storage conditions (e.g., high temperature or high humidity), the anserine content in the composition decreases. In particular, when an anserine-containing composition is used as an oral composition such as a health food, even if a desired amount of anserine is incorporated into the composition, the anserine content decreases over time due to long-term storage under high temperature or high humidity conditions, making it difficult to ingest the desired amount of anserine. Furthermore, the decrease in anserine content is problematic from the perspective of maintaining the quality of oral compositions such as health foods. Furthermore, if the anserine content in the composition decreases over time, the effects achieved by incorporating anserine in the composition as described in Patent Documents 1 and 2 may also decrease over time.

[0005] For these reasons, there is a need for a method to improve the stability of anserine in compositions. Patent Documents 1 and 2 make no mention of the property of anserine that it decomposes due to heat or humidity and decreases over time, and do not consider at all the stability of anserine itself.

[0006] An object of the present invention is to provide a method for suppressing the decrease (decomposition) of anserine over time due to heat and improving the stability of anserine. [Means for solving the problem]

[0007] As a result of extensive research, the present inventors have discovered that the decrease (decomposition) of anserine over time due to heat can be suppressed and the stability of anserine can be improved by mixing anserine with quercetin or a glycoside thereof so that the weight ratio of quercetin or a glycoside thereof to anserine in the composition, calculated as isoquercitrin (quercetin or a glycoside thereof / anserine calculated as isoquercitrin), is 5 or more, thereby completing the present invention.

[0008] The present invention encompasses the following methods, uses and compositions. [1] A method for improving the stability of anserine, comprising a step of mixing anserine with quercetin or a glycoside thereof so that the weight ratio of quercetin or a glycoside thereof to anserine in the composition, converted into isoquercitrin (quercetin or a glycoside thereof / anserine converted into isoquercitrin) is 5 or more. [2] The method for improving the stability of anserine according to [1] above, wherein quercetin or a glycoside thereof is a quercetin glycoside. [3] The method for improving the stability of anserine according to [1] or [2] above, wherein the composition is an oral composition. [4] The method for improving the stability of anserine according to any one of [1] to [3] above, wherein the composition is a food or drink. [5] The method for improving the stability of anserine according to any one of [1] to [4] above, wherein the composition is a solid formulation. [6] The method for improving the stability of anserine according to [5] above, wherein the solid preparation is a tablet, granule, or powder. [7] Use of quercetin or its glycoside to improve the stability of anserine, by mixing anserine with quercetin or its glycoside so that the weight ratio of quercetin or its glycoside to anserine in the composition, converted into isoquercitrin (quercetin or its glycoside / anserine converted into isoquercitrin), is 5 or more. [8] A composition containing anserine and quercetin or its glycoside, wherein the weight ratio of quercetin or its glycoside to anserine in terms of isoquercitrin (quercetin or its glycoside / anserine in terms of isoquercitrin) is 5 or more, and the stability of anserine is improved. [Effects of the Invention]

[0009] According to the present invention, the stability of anserine can be improved by suppressing the decrease (decomposition) of anserine over time due to heat. This makes it possible to suppress the decrease in anserine in a composition containing anserine (particularly an oral composition such as a health food) over time, even when the composition is stored for a long period of time under inappropriate storage conditions (e.g., high-temperature conditions). Furthermore, the quality of the oral composition such as a health food is maintained. The present invention also provides the use of quercetin or a glycoside thereof to improve the stability of anserine, and a composition having improved anserine stability. DETAILED DESCRIPTION OF THE INVENTION

[0010] The method of the present invention for improving the stability of anserine (hereinafter also simply referred to as the method of the present invention) comprises the step of mixing anserine with quercetin or a glycoside thereof so that the weight ratio of quercetin or a glycoside thereof to anserine in the composition, calculated as isoquercitrin (quercetin or a glycoside thereof / anserine calculated as isoquercitrin) is 5 or more. By mixing anserine with quercetin or a glycoside thereof so that the weight ratio of quercetin or a glycoside thereof to anserine in the composition, calculated as isoquercitrin (quercetin or a glycoside thereof / anserine calculated as isoquercitrin) is 5 or more, it is possible to suppress the decrease (decomposition) of anserine over time due to heat and improve the stability of anserine.

[0011] Anserine is a type of imidazole peptide, and is a compound that has a structure in which a methyl group is bonded to the 1-position of the L-histidine residue in carnosine, a dipeptide formed by the peptide bond between β-alanine and L-histidine. Anserine decomposes under heat (e.g., about 40°C) and humidity, and decreases over time.

[0012] Imidazole peptides are a general term for amino acid conjugates containing an imidazole group. Imidazole is a nitrogen-containing heterocyclic aromatic compound containing nitrogen atoms at the 1- and 3-positions of a five-membered ring. Imidazole peptides such as anserine are widely distributed in the human body and are particularly abundant in the muscles of vertebrates, making them useful in health foods designed to prevent muscle weakness. Imidazole peptides such as anserine are found in the muscles of tuna, bonito, salmon, and other fish, particularly those required for endurance, such as migratory birds and tuna and bonito, where they function as major components of muscle. It has also been reported that imidazole peptides decrease with age (Stuerenburg et al., Arch. Gerontol. Geriatr. 1999). Therefore, the amount of imidazole peptides lost through normal metabolism must be supplemented externally by various methods.

[0013] In the method of the present invention, the origin or raw material of anserine is not particularly limited, and anserine-containing material extracted with an aqueous solvent, chemically synthesized anserine, anserine from which protein has been removed (protein-removed product), etc. can be used. Among these, anserine-containing material extracted with an aqueous solvent is preferred. An anserine-containing material extracted with an aqueous solvent is an anserine-containing material extracted using an aqueous solvent from a material containing anserine (e.g., fish meat).

[0014] When extracting from a material containing anserine using an aqueous solvent, the material is not particularly limited and can be selected appropriately depending on the desired efficacy and application, but since anserine is found in high concentrations in the muscles of migratory fish such as tuna and bonito, it is preferable to use migratory fish, and it is more preferable to use tuna, bonito, etc. The part of the migratory fish used as the material is also not particularly limited, and any desired part can be used as long as it contains anserine.

[0015] Water is preferred as the aqueous solvent. The temperature of the water used for extraction is not particularly limited, and water of various temperatures (hot water, warm water, room temperature water) can be used. The aqueous solvent-extracted anserine-containing product can be obtained as a powder by, for example, separating oil from the broth of tuna, bonito, etc., purifying it by methods such as bleaching, deodorizing, desalting, and concentrating it, adding an excipient such as dextrin, and spray-drying it.

[0016] Commercially available products that can be used as anserine include, for example, Marine Active 10 (aqueous solvent-extracted anserine-containing product, manufactured by Yaizu Suisan Kagaku Co., Ltd.) and SA-30(IK)SD (anserine-extracted product from salmon, manufactured by Tokai Bussan Co., Ltd.).

[0017] In the method of the present invention, the content of anserine in the composition is not particularly limited, but is preferably 0.01 wt% or more, more preferably 0.05 wt% or more, even more preferably 0.1 wt% or more, and is preferably 16.6 wt% or less, more preferably 3.0 wt% or less, even more preferably 1.0 wt% or less, and particularly preferably 0.7 wt% or less. In one embodiment, the content of anserine in the composition is preferably 0.01 to 16.6 wt%, more preferably 0.05 to 3.0 wt%, even more preferably 0.1 to 1.0 wt%, and particularly preferably 0.1 to 0.7 wt%. The anserine content can be measured, for example, by the anserine quantification method described in the Examples of the present application.

[0018] In the method of the present invention, when the composition is an oral composition, the anserine content in the intake (administration) amount of the composition per adult per day per 60 kg body weight is not particularly limited, but is preferably 1 to 100 mg, more preferably 5 to 20 mg.

[0019] As used herein, "quercetin" refers to a compound that belongs to the flavonol class, a type of polyphenol. As used herein, "quercetin glycoside" refers to a glycoside of quercetin, specifically a general term for a series of compounds in which one or more (e.g., 1 to 10) sugars are glycosidically bonded to the hydroxyl group at the 3-position and / or the hydroxyl group at the 4'-position of quercetin. Quercetin glycosides may be one type of compound or two or more types of compounds.

[0020] In one embodiment, the quercetin glycoside is preferably a compound in which one or more sugars are glycosidically bonded to the hydroxyl group at the 3-position of quercetin. The quercetin glycoside in which one or more sugars are glycosidically bonded to the hydroxyl group at the 3-position of quercetin is a compound represented by the following general formula: (X) n represents a sugar chain, X represents a sugar (monosaccharide), and n is an integer of 1 or more (for example, 1 to 10).

[0021] [ka]

[0022] The sugar (monosaccharide) constituting the sugar chain represented by X glycosidically bonded to quercetin is, for example, glucose, rhamnose, galactose, glucuronic acid, etc., and preferably glucose or rhamnose. The sugar bonded to quercetin may be modified (for example, malonylated). Furthermore, n is not particularly limited as long as it is 1 or more, but is preferably 1 to 16, more preferably 1 to 13, and even more preferably 1 to 10. When n is 2 or more, the X moiety may consist of one type of sugar or multiple types of sugars. In other words, when n is 2 or more, (X) n may be a sugar chain consisting of one type of sugar, or may be a sugar chain consisting of multiple types of sugar.

[0023] In the method of the present invention, the quercetin glycoside also includes a quercetin glycoside obtained by treating an existing quercetin glycoside with an enzyme or the like to undergo glycosylation. Examples of quercetin glycosides in the method of the present invention include rutin (quercetin 3-rutinoside), enzyme-treated rutin (enzyme-treated rutin), quercitrin (quercetin-3-O-rhamnoside), isoquercitrin (quercetin 3-glucoside), quercetin 3,4'-diglucoside, quercetin 4'-glucoside, quercetin 3-(6-malonylglucoside), hyperoside (quercetin 3-galactoside), and quercetin-3-O-glucuronide. Enzyme-treated rutin is a quercetin glycoside in which a sugar chain is further bound to the glucosidic moiety of isoquercitrin. Preferred examples of enzymatically treated rutin include isoquercitrin obtained by enzymatically treating rutin to remove the rhamnose sugar chain moiety and then treating the resulting isoquercitrin with a glycosyltransferase, to which a sugar chain consisting of 1 to 7 glucose units is bound, and mixtures thereof as the main component. In the method of the present invention, quercetin or a glycoside thereof is preferably a quercetin glycoside, and more preferably at least one selected from the group consisting of isoquercitrin and glycosides in which a sugar chain consisting of 1 to 9 glucose units is bound to isoquercitrin.

[0024] As quercetin or its glycosides, one compound may be used alone, or two or more compounds may be used in combination. When two or more compounds are used in combination as quercetin or its glycosides, the content of quercetin or its glycosides is the total content of the two or more compounds.

[0025] In the method of the present invention, there are no particular limitations on the origin or production method for obtaining quercetin or its glycosides. For example, plants that are known to contain large amounts of quercetin or its glycosides include buckwheat, Sophora japonica, capers, apples, tea, onions, grapes, broccoli, mulukhiyah, raspberries, cowberries, cranberries, opuntia, rafuma, leafy vegetables, and citrus fruits, and quercetin or its glycosides can be obtained from these plants. In the method of the present invention, quercetin or a glycoside thereof can be obtained by concentrating or purifying an extract derived from a natural product such as the above-mentioned plant to increase the content of quercetin or a glycoside thereof, as long as the effects of the present invention are not impaired. For example, a concentrate or purified product of an extract containing quercetin or a glycoside thereof can be used. Known methods can be used for concentration or purification. In the method of the present invention, purified or isolated quercetin or a glycoside thereof may be used. Raw materials containing quercetin or a glycoside thereof may also be used. Chemically synthesized quercetin or a glycoside thereof can also be used.

[0026] The content of quercetin or its glycoside in a raw material containing quercetin or its glycoside is not particularly limited, but is preferably 10 to 99% by weight, more preferably 15 to 50% by weight, and more specifically, may be, for example, 15%, 30%, or 40% by weight. Commercially available products that can be used as quercetin or its glycoside include, for example, San-Emic (manufactured by San-Ei Gen F.F.I., Inc.).

[0027] In the method of the present invention, the content of quercetin or its glycosides in the composition, calculated as isoquercitrin, is not particularly limited, but is preferably 0.05% by weight or more, more preferably 0.25% by weight or more, even more preferably 0.5% by weight or more, even more preferably 1.5% by weight or more, and particularly preferably 2% by weight or more, and is preferably 83.4% by weight or less, more preferably 60% by weight or less, even more preferably 20% by weight or less, even more preferably 14% by weight or less, and particularly preferably 10% by weight or less. In one embodiment, the content of quercetin or its glycosides in the composition, calculated as isoquercitrin, may be preferably 0.05 to 83.4% by weight, more preferably 0.25 to 60% by weight, even more preferably 0.5 to 20% by weight, even more preferably 1.5 to 14% by weight, and particularly preferably 2 to 10% by weight.

[0028] The content of quercetin or its glycosides in terms of isoquercitrin can be measured, for example, by high performance liquid chromatography (HPLC). Specifically, for example, peaks derived from quercetin or its glycosides are detected by HPLC, and the content in terms of isoquercitrin can be calculated from the total area of these peaks. Regarding quercetin or its glycosides, the amount converted to isoquercitrin or similar expressions means the amount of isoquercitrin. In the case of quercetin or a quercetin glycoside other than isoquercitrin, it means the value obtained by multiplying the number of moles of the compound by the molecular weight of isoquercitrin.

[0029] In the method of the present invention, when the composition is an oral composition, the content of quercetin or its glycoside in the daily intake (administration) of the composition per adult per 60 kg body weight is not particularly limited, but is preferably 1 to 1000 mg, more preferably 10 to 500 mg, in terms of isoquercitrin.

[0030] The method of the present invention comprises a step of mixing quercetin or a glycoside thereof to anserine in the composition so that the weight ratio, in terms of isoquercitrin, of quercetin or a glycoside thereof (quercetin or a glycoside thereof in terms of isoquercitrin / anserine) is 5 or more. That is, the method comprises a step of mixing these components so that the weight ratio, in terms of isoquercitrin, of the content of quercetin or a glycoside thereof to the content of anserine in the resulting composition (quercetin or a glycoside thereof in terms of isoquercitrin / anserine) is 5 or more. The above step can be carried out by mixing anserine and quercetin or a glycoside thereof so that the weight ratio, in terms of isoquercitrin, of quercetin or a glycoside thereof to the weight of anserine (quercetin or a glycoside thereof / anserine) is 5 or more. The weight ratio of quercetin or a glycoside thereof to anserine in terms of isoquercitrin (quercetin or a glycoside thereof in terms of isoquercitrin / anserine) is preferably 5.1 or more, more preferably 5.2 or more, and even more preferably 9.5 or more. The greater the weight ratio of quercetin or a glycoside thereof to anserine in terms of isoquercitrin (quercetin or a glycoside thereof in terms of isoquercitrin / anserine), the greater the effect of improving the stability of anserine in the composition. Therefore, the upper limit is not particularly limited, but is, for example, 20 or less, preferably 18 or less. In one aspect, the weight ratio of quercetin or a glycoside thereof to anserine in terms of isoquercitrin (quercetin or a glycoside thereof in terms of isoquercitrin / anserine) may be 5 to 20, preferably 5.1 to 18, more preferably 5.2 to 18, and even more preferably 9.5 to 18. In the method of the present invention, anserine and quercetin or its glycosides, calculated as isoquercitrin, are preferably used in the weight ratio described above. When the weight ratio of the content of quercetin or its glycosides, calculated as isoquercitrin, to the content of anserine in the composition (quercetin or its glycosides, calculated as isoquercitrin / anserine) is within the above range, the decrease (decomposition) of anserine over time due to heat can be suppressed, and the stability of anserine can be improved. According to the method of the present invention, the stability of anserine in the composition can be improved.

[0031] In the method of the present invention, the composition is obtained by mixing anserine and quercetin or a glycoside thereof so that the weight ratio of quercetin or a glycoside thereof to anserine in the composition, calculated as isoquercitrin (quercetin or a glycoside thereof / anserine calculated as isoquercitrin), is equal to or greater than a specific value, thereby suppressing the decrease (decomposition) of anserine over time due to heat and improving the stability of anserine. The composition in the method of the present invention is useful as a composition (preferably an oral composition such as a health food) with improved anserine stability.

[0032] In the method of the present invention, the anserine retention rate of the composition after one month of storage, for example, the ratio of the anserine content in the composition after one month of storage under conditions of 40°C and a relative humidity (RH) of 75% to the anserine content in the composition immediately after preparation ((anserine content in the composition after one month of storage / anserine content in the composition immediately after preparation) x 100), is preferably 90% or more, more preferably 93% or more, and even more preferably 95% or more. The upper limit of the anserine retention rate after one month of storage under the above conditions is not particularly limited, but may be, for example, 100%.

[0033] In the method of the present invention, the composition may be in the form of, for example, a food or drink, a pharmaceutical product, a quasi-drug, or a feed. The composition may itself be a food or drink, a pharmaceutical product, a quasi-drug, or a feed, or may be a material or preparation to be incorporated into these. In one embodiment, when the composition is a food or drink, a pharmaceutical product, a quasi-drug, or a feed, the content of anserine in the composition is preferably 0.01 to 16.6 wt%, more preferably 0.05 to 3.0 wt%, even more preferably 0.1 to 1.0 wt%, and particularly preferably 0.1 to 0.7 wt%. The content of quercetin or its glycoside in the composition, calculated as isoquercitrin, may be preferably 0.05 to 83.4 wt%, more preferably 0.25 to 60 wt%, even more preferably 0.5 to 20 wt%, even more preferably 1.5 to 14 wt%, and particularly preferably 2 to 10 wt%.

[0034] In one embodiment, the composition used in the method of the present invention is preferably an oral composition. Examples of oral compositions include foods and beverages, oral medicines, quasi-drugs, feeds, etc., and are preferably foods and beverages or oral medicines, and more preferably foods and beverages. In one embodiment, the composition in the method of the present invention is useful as a composition having improved stability of anserine, for example, as a food or drink, oral pharmaceutical product, oral quasi-drug, feed, etc. having improved stability of anserine.

[0035] The composition used in the method of the present invention may contain any additives and any components in addition to anserine and quercetin or its glycoside, as long as the effects of the present invention are not impaired. These additives and components can be selected depending on the form of the composition, and those that can generally be used in foods and beverages, pharmaceuticals, quasi-drugs, feed, etc. can be used. When the composition is used as a food and beverage, pharmaceutical, quasi-drug, feed, etc., the production method thereof is not particularly limited, and it can be produced by a general method.

[0036] In one embodiment, the composition used in the method of the present invention may contain, in addition to anserine and quercetin or glycosides thereof, other materials with in vivo functionality. Examples of such materials include imidazole peptides other than anserine, glucosamines, chondroitin sulfate, proteoglycans, collagens such as type II collagen, hyaluronic acid, and vitamins such as vitamin D. These materials may be used alone or in combination.

[0037] In one embodiment, the composition used in the method of the present invention may contain an imidazole peptide other than anserine, such as carnosine, balenine, homoserine, etc.

[0038] The content of imidazole peptides other than anserine in the composition is not particularly limited, and can be appropriately adjusted so that the intake of imidazole peptides other than anserine per individual per day is, for example, 1 to 1500 mg, preferably 2.5 to 100 mg. Furthermore, the intake per kg of body weight can be, for example, 0.02 to 30.0 mg / kg, preferably 0.05 to 2.00 mg / kg. For example, when the composition used in the method of the present invention is formulated as a food or beverage, the content of imidazole peptides other than anserine is, for example, 0.01 to 95% by weight, preferably 0.05 to 1% by weight, and more preferably 0.1 to 0.5% by weight.

[0039] In one embodiment, the composition used in the method of the present invention may contain glucosamines. Glucosamines are 2-aminoglucoses, in which the hydroxyl group at the 2-position of glucose is replaced with an amino group. They are natural amino sugars widely distributed in nature as constituent sugar molecules of biological components such as glycoproteins, glycolipids, and mucopolysaccharides. In recent years, glucosamines have not only been recognized as important basic constituent molecules of biological components, but also have been shown to have various beneficial effects when ingested. Consequently, they are widely used in health foods for the treatment and prevention of joint pain and osteoarthritis. It is desirable to incorporate glucosamines together with anserine and quercetin or their glycosides in the composition used in the method of the present invention. In this specification, glucosamines also include their salts.

[0040] The glucosamine content of the composition is not particularly limited and can be adjusted appropriately so that the glucosamine intake per individual per day is, for example, 100 to 5,000 mg, preferably 300 to 2,000 mg. The intake per kg of body weight can be, for example, 2 to 100 mg / kg, preferably 6.0 to 40.0 mg / kg. For example, when the composition of the present invention is formulated as a solid preparation, the glucosamine content can be, for example, 0.1 to 95% by weight, preferably 10 to 80% by weight, of the total solid preparation.

[0041] In one embodiment, the composition used in the method of the present invention may contain chondroitin sulfate. Chondroitin sulfate is widely distributed in the human body, and is particularly abundant in the cartilage and skin of joints, and is therefore used in pharmaceuticals and health foods for the purposes of improving osteoarthritis and improving skin beauty. In this specification, chondroitin sulfate also includes its salts.

[0042] The content of chondroitin sulfate in the composition is not particularly limited, and can be appropriately adjusted so that the intake of chondroitin sulfate is, for example, 10 to 3,000 mg, preferably 50 to 1,000 mg per individual per day. Furthermore, the intake per kg of body weight can be, for example, 0.2 to 60.0 mg / kg, preferably 1.0 to 20.0 mg / kg. For example, when the composition in the method of the present invention is formulated as a solid preparation, the content of chondroitin sulfate can be, for example, 0.1 to 95% by weight, preferably 0.5 to 20% by weight, based on the total weight of the solid preparation.

[0043] In one embodiment, the composition used in the method of the present invention may contain vitamin D. Vitamin D is not only important as a basic structural molecule of biological components, but its ingestion has also been confirmed to have various beneficial effects, and it is expected to have the potential to suppress muscle weakness. Vitamin D includes vitamin D2 produced by plants and vitamin D3 produced by animals. Vitamin D3 is known to be effective in humans and can be obtained industrially by separating and purifying it from wool fat. In this specification, vitamin D includes vitamin D2 and vitamin D3.

[0044] The vitamin D content in the composition is not particularly limited, and can be appropriately adjusted so that the vitamin D intake per individual per day is, for example, 0.5 to 50 μg, preferably 1 to 25 μg. The intake per kg of body weight can be, for example, 0.01 to 1 μg / kg, preferably 0.02 to 0.5 μg / kg. For example, when the composition in the method of the present invention is used as a food or drink, the vitamin D content can be, for example, 0.00001 to 95% by weight, preferably 0.00005 to 10% by weight, more preferably 0.0001 to 0.001% by weight of the total composition.

[0045] In the method of the present invention, when anserine and quercetin or its glycoside are mixed in the above weight ratio, any additives or components may be mixed in addition to them. In the method of the present invention, the method, conditions, etc. for mixing anserine, quercetin or its glycoside, and the above-mentioned optional additives and optional components are not particularly limited, and methods, conditions, etc. known in the technical field can be appropriately adopted.

[0046] The method of the present invention may include a step other than the step of mixing anserine with quercetin or a glycoside thereof. For example, the method of the present invention may include a step of formulating the composition obtained in the step of mixing anserine with quercetin or a glycoside thereof into a desired dosage form (e.g., a solid formulation as described below). For formulation, methods known in the art can be used.

[0047] In the method of the present invention, the form of the composition is not particularly limited and may be a solid, liquid, or the like, but a solid is preferred. When the composition is a solid, the decrease in anserine over time due to heat can be more effectively suppressed. Examples of solid formulations include tablets, granules, powders, fine granules, powders, capsules, chewable tablets, and tablets. The solid formulation is preferably a tablet, granule, or powder. Additives such as excipients, lubricants, binders, and disintegrants can be used in the solid formulation as desired. Examples of such additives include crystalline cellulose, dextrin, silicon oxide, and calcium stearate. Examples of liquid formulations include syrups, suspensions, and emulsions.

[0048] In the method of the present invention, when the composition is prepared as a food or beverage, for example, anserine and quercetin or its glycoside can be mixed with ingredients usable in foods and beverages (e.g., food ingredients, food additives used as needed, etc.) to prepare various foods and beverages. The foods and beverages are not particularly limited, and examples include general foods and beverages, health foods, health drinks, functional foods, foods for specified health uses, health supplements, and foods and beverages for medical patients. Health foods, functional foods, foods for specified health uses, health supplements, and foods and beverages for medical patients can be prepared in various dosage forms, such as tablets, granules, powders, fine granules, powders, capsules, chewable tablets, tablets, syrups, and liquid diets. Among these, tablets, granules, and powders are preferred.

[0049] In the method of the present invention, when the composition is to be prepared as a pharmaceutical or quasi-drug, for example, anserine and quercetin or its glycoside are mixed together with a pharmacologically acceptable carrier and, if necessary, additives, etc., to prepare the pharmaceutical or quasi-drug in various dosage forms. Such carriers, additives, etc. may be any pharmacologically acceptable carriers that can be used in pharmaceuticals or quasi-drugs, and may include, for example, one or more of excipients, binders, disintegrants, lubricants, antioxidants, colorants, etc. The pharmaceutical or quasi-drug may be administered orally or parenterally, with oral administration being preferred. When the composition is to be prepared as a pharmaceutical or quasi-drug, an oral pharmaceutical or quasi-drug is preferred. Dosage forms for oral administration include tablets, granules, powders, fine granules, powders, capsules, chewable tablets, tablets, sugar-coated tablets, suspensions, emulsions, etc. Among these, tablets, granules, or powders are preferred. Dosage forms for parenteral administration include injections and infusions.

[0050] In the method of the present invention, when the composition is used as a feed, for example, anserine and quercetin or its glycoside may be mixed with the feed. Feed also includes feed additives. Examples of feed include livestock feed for cattle, pigs, chickens, sheep, horses, etc.; small animal feed for rabbits, rats, mice, etc.; and pet food for dogs, cats, small birds, etc.

[0051] In the method of the present invention, when the composition is an oral composition, the subject to which the composition is ingested or administered (also referred to as the administration subject) is not particularly limited, and is preferably a human or a non-human mammal, more preferably a human.

[0052] In the method of the present invention, the stability of anserine can be improved by mixing anserine with quercetin or a glycoside thereof so that the weight ratio of quercetin or a glycoside thereof to anserine in the composition, calculated as isoquercitrin (quercetin or a glycoside thereof / anserine calculated as isoquercitrin) is at least 5. The method of the present invention for improving anserine stability can also be referred to as a method for improving anserine stability or a method for stabilizing anserine.

[0053] Anserine, quercetin, or their glycosides are compounds that are contained in natural products and foods and beverages and have been consumed as food. Therefore, from the viewpoint of safety, anserine, quercetin, or their glycosides are thought to pose few problems even when ingested daily.

[0054] In one embodiment, the composition in the method of the present invention can be used, for example, by adding it to an oral composition. In one embodiment, the composition in the method of the present invention can be used as a food additive. The oral composition to which the composition in the method of the present invention is added is not particularly limited, and examples include foods and beverages, oral pharmaceuticals, oral quasi-drugs, feeds, etc., and preferably foods and beverages.

[0055] The present invention also encompasses the following uses and compositions (hereinafter, also simply referred to as the use of the present invention and the composition of the present invention, respectively). Use of quercetin or its glycoside to improve the stability of anserine, which comprises mixing anserine with quercetin or its glycoside so that the weight ratio of quercetin or its glycoside to anserine in the composition, converted into isoquercitrin (quercetin or its glycoside / anserine converted into isoquercitrin), is 5 or more. A composition containing anserine and quercetin or its glycoside, wherein the weight ratio of quercetin or its glycoside to anserine in terms of isoquercitrin (quercetin or its glycoside / anserine in terms of isoquercitrin) is 5 or more, and the stability of anserine is improved.

[0056] Use of the present invention makes it possible to inhibit the decrease (decomposition) of anserine in a composition over time due to heat, thereby improving the stability of anserine. Furthermore, the composition of the present invention inhibits the decrease (decomposition) of anserine in a composition over time due to heat, thereby improving the stability of anserine. By improving the stability of anserine, it becomes easier to ingest a desired amount of anserine, even when a composition containing anserine (particularly an oral composition such as a health food) is stored for a long period of time under inappropriate storage conditions (e.g., high-temperature conditions). In the use and composition of the present invention, anserine, quercetin, or its glycoside, preferred embodiments thereof, and the preferred range of the weight ratio of quercetin or its glycoside to anserine in the composition in terms of isoquercitrin (quercetin or its glycoside in terms of isoquercitrin / anserine) are the same as those in the method of the present invention described above. In addition, optional additives, optional ingredients, and the form of the composition are also the same as those in the method of the present invention. For example, the weight ratio of quercetin or its glycoside to anserine in terms of isoquercitrin (quercetin or its glycoside in terms of isoquercitrin / anserine) may be 5 to 20, preferably 5.1 to 18, more preferably 5.2 to 18, and even more preferably 9.5 to 18.

[0057] In this specification, a numerical range expressed by a lower limit and an upper limit, i.e., "lower limit to upper limit," includes both the lower limit and the upper limit. For example, a range expressed by "1 to 2" means 1 or more and 2 or less, including 1 and 2. In this specification, the upper and lower limits may be any combination of ranges. All academic literature and patent documents described in this specification are incorporated herein by reference. [Example]

[0058] The present invention will be described in more detail below with reference to examples, although the present invention is not limited to these examples.

[0059] (Examples 1 to 4, Comparative Examples 1 to 3) <Raw materials used> In Examples 1 to 4 and Comparative Examples 1 to 3, the following raw materials were used. Anserine raw material (Marine Active 10, aqueous solvent extract containing anserine, containing 10% anserine by weight, manufactured by Yaizu Suisan Kagaku Co., Ltd.) Quercetin glycoside raw material (Sanemic, manufactured by San-Ei Gen F.F.I. Co., Ltd., quercetin glycoside isoquercitrin content: 11.4% by weight) Cellulose raw material (Ceolas ST-100, manufactured by Asahi Kasei Corporation)

[0060] The raw materials were mixed in the weights shown in Table 2 below to prepare 200 g samples of each composition (mixed powder). A portion (50 g) of the composition immediately after preparation was subjected to the following method for quantifying anserine. The anserine content (g), quercetin or its glycoside content (converted to isoquercitrin) (g), quercetin or its glycoside content (converted to isoquercitrin) (wt%), and the weight ratio of the quercetin or its glycoside content (converted to isoquercitrin) to the anserine content immediately after preparation (quercetin or its glycoside (converted to isoquercitrin) / anserine) are shown in Table 2. The remaining composition was placed in an aluminum pouch (manufactured by Seisan Nippon Co., Ltd.), sealed, and allowed to stand (store) in a thermostatic chamber (manufactured by Espec Corporation) at 40°C and a relative humidity (RH) of 75% for one month. After one month of storage, 50 g of the composition was removed from the aluminum pouch, and the anserine content was again quantified using the method described below. The anserine content immediately after preparation was defined as 100%, and the residual anserine rate (%) after one month of storage was calculated. The results are shown in Table 2. In Table 2, the residual anserine rate (%) after one month of storage represents the ratio of the anserine content in the composition after one month of storage to the anserine content in the composition immediately after preparation ((anserine content in the composition after one month of storage / anserine content in the composition immediately after preparation) × 100).

[0061] <Method for quantifying anserine> 1. Preparation of sodium citrate buffer (pH 2.2) To prepare a sodium citrate buffer solution, 980 g of trisodium citrate dihydrate was dissolved in approximately 3500 mL of water, and approximately 700 mL of hydrochloric acid and 5 mL of octanoic acid were added. The pH was adjusted to 2.2 with hydrochloric acid, and then the resulting solution was diluted to 5000 mL with water. 100 mL of thiodiethylene glycol and 4000 mL of water were added to 500 mL of the sodium citrate buffer solution, and the pH was adjusted to 2.2 with hydrochloric acid, and then the resulting solution was diluted to 5000 mL with water to obtain a sodium citrate buffer solution (pH 2.2).

[0062] 2. Preparation of Standard Solutions L-anserine nitrate standard material (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd., 77.3-81.2% as anserine) was dissolved in 0.01 mol / L hydrochloric acid to prepare a 2.5 μmol / mL standard stock solution, which was then further diluted with sodium citrate buffer (pH 2.2) to 0.5 μmol / mL to prepare the standard solution.

[0063] 3. Preparation of Test Solutions The compositions (mixed powders) obtained in each Example and Comparative Example were milled, and approximately 1 g of each was precisely weighed out. 25 mL of 10% sulfosalicylic acid solution was added and the mixture was shaken for 20 minutes. The pH was adjusted to approximately 2.2 with sodium hydroxide solution, and the solution was diluted to 50 mL with sodium citrate buffer (pH 2.2). The solution was then filtered through filter paper (No. 1) (manufactured by Toyo Roshi Kaisha, Ltd.). The resulting solution was filtered through a membrane filter with a pore size of 0.45 μm, and the resulting solution was used as the test solution.

[0064] 4. Measurement 20 μL of each of the standard solution and the test solution was injected into an automatic amino acid analyzer, and anserine in the test solution was identified (qualified) from the peak retention time of anserine in the standard solution. The peak height was measured, and the anserine content was calculated using the following formula. Anserine content (g / 100g) = S x 240.26 x A / B x V x 100 / W x 10 -6 S: concentration of standard solution (μmol / mL) A: Peak height of the test solution B: Peak height of standard solution V: fixed volume of test solution (mL) W: Sample volume (g) 240.26: Molecular weight of anserine

[0065] Conditions for automatic amino acid analysis Equipment: L-8900 high-speed amino acid analyzer (Hitachi High-Tech Corporation) Column: Hitachi custom ion exchange resin, φ4.6 mm x 60 mm (Hitachi High-Tech Corporation) Mobile phase: Buffer solution for biological fluid analysis PF KANTO (PF-1 to PF-RG) (Kanto Chemical Co., Ltd.) Reaction solution: Hitachi ninhydrin coloring solution kit (Fujifilm Wako Pure Chemical Industries, Ltd.) Flow rate: Mobile phase 0.35 mL / min, reaction solution 0.30 mL / min Reactor temperature: 135℃ Measurement wavelength: 570nm Automated amino acid analyzer time program: See Table 1 below

[0066] [Table 1]

[0067] [Table 2]

Claims

1. A method for improving the stability of anserine, comprising a step of mixing anserine with quercetin or a glycoside thereof so that the weight ratio of quercetin or a glycoside thereof to anserine in the composition, calculated as isoquercitrin (quercetin or a glycoside thereof / anserine calculated as isoquercitrin) is 5 or more.

2. 2. The method for improving the stability of anserine according to claim 1, wherein quercetin or a glycoside thereof is a quercetin glycoside.

3. 3. The method for improving the stability of anserine according to claim 1 or 2, wherein the composition is an oral composition.

4. 3. The method for improving the stability of anserine according to claim 1 or 2, wherein the composition is a food or drink.

5. 3. The method for improving the stability of anserine according to claim 1 or 2, wherein the composition is a solid formulation.

6. 6. The method for improving the stability of anserine according to claim 5, wherein the solid preparation is a tablet, granule, or powder.

7. Use of quercetin or a glycoside thereof for improving the stability of anserine, which comprises mixing anserine with quercetin or a glycoside thereof so that the weight ratio of quercetin or a glycoside thereof to anserine in the composition, calculated as isoquercitrin (quercetin or a glycoside thereof / anserine calculated as isoquercitrin), is 5 or more.

8. A composition containing anserine and quercetin or a glycoside thereof, in which the weight ratio of quercetin or a glycoside thereof to anserine in terms of isoquercitrin (quercetin or a glycoside thereof / anserine in terms of isoquercitrin) is 5 or more, and in which the stability of anserine is improved.

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