Method for producing an ether-type glycerophospholipid-containing functional material

Abstract

To provide a method for producing efficiently and in large amount a functional material containing a large amount of ether-type glycerophospholipids, which reduces an amount of a solvent used at the time of extraction and is easy to scale up.SOLUTION: Provided is a method for producing a functional material containing ether-type glycerophospholipids, comprising performing an extraction treatment using an ethanol / hexane mixture on biological materials such as bivalve tissues which have been subjected to a drying treatment.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] This invention relates to a method for producing a functional material containing ether-type glycerophospholipid. More specifically, it relates to a method for producing a functional material containing ether-type glycerophospholipid, which is extremely advantageous in terms of economy by reducing the amount of extraction solvent used.

Background Art

[0002] Glycerophospholipids are known to be important as components of biological membranes. This glycerophospholipid can be classified into subclasses of diacyl-type glycerophospholipid, alkenylacyl-type glycerophospholipid (plasmalogen), and alkyl ether-type glycerophospholipid.

[0003] Among the above glycerophospholipids, alkenylacyl-type glycerophospholipid (plasmalogen) and alkyl ether-type phospholipid have an ether bond, and thus are collectively called ether-type glycerophospholipid.

[0004] Among them, plasmalogen having a vinyl ether bond at the 1-position of the fatty acid is a phospholipid that is characteristically abundant in brain nerve cells and myocardium, and is a lipid component that has attracted attention in recent years.

[0005] This plasmalogen has a biological function characteristic of the vinyl ether structure in the molecule, scavenges active oxygen, radicals, and metal ions to exhibit antioxidant properties, and is also reported to be involved in the fluid flexibility of cell membranes (especially nerve cell synapse membranes) (Non-Patent Document 1).

[0006] Furthermore, it has also been proposed to improve and prevent diseases such as Alzheimer's disease by including plasmalogen or ether-type glycerophospholipid in food and drink products and pharmaceuticals (Patent Documents 1 to 4).

[0007] As a specific method for obtaining such plasmalogens or ether-type glycerophospholipids, a method of extracting the plasmalogens or ether-type glycerophospholipids from various raw materials using an extraction solvent is known.

[0008] For example, in Japanese Patent Application Laid-Open No. 2007-262024 (Patent Document 5), a method for extracting plasmalogen-containing lipids from aquatic animals using only solvents that can be used in the food and beverage field, and which can be incorporated into, for example, functional foods and beverages, has been proposed.

[0009] This method for extracting plasmalogen-containing lipids comprises an extraction step of adding a mixed solvent of n-hexane / ethanol / water to a dried product of an aquatic animal, stirring, and recovering the filtrate, a liquid-liquid separation step of adding a mixed solvent of n-hexane / water to the filtrate recovered in the above extraction step, stirring, allowing to stand, and recovering the upper layer, and a solvent removal step of removing the solvent from the upper layer recovered in the above liquid-liquid separation step and recovering the lipid.

[0010] Furthermore, in Japanese Patent Application Laid-Open No. 2010-65167 (Patent Document 6), a method for producing plasmalogen-type phospholipids and sphingolipids from animal tissues, which can produce high-purity plasmalogen-type phospholipids and sphingolipid fractions using animal tissues that can be obtained inexpensively and in large quantities, has been proposed.

[0011] This method for producing plasmalogen-type phospholipids and sphingolipids (A) a step of subjecting an animal tissue containing plasmalogen-type phospholipids and sphingolipids to ethanol extraction treatment to obtain an ethanol extract, (B) a step of hydrolyzing the diacyl-type glycerophospholipids contained in the ethanol extract obtained in the step (A), (C) a step of treating the treated product obtained in the step (B) with a water-soluble ketone solvent and recovering the insoluble portion, Step (D): subjecting the insoluble matter obtained in step (C) to solvent partitioning with a mixed organic solvent of an aliphatic hydrocarbon solvent and a water-soluble ketone solvent, and water, and recovering the mixed organic solvent portion which includes the following

[0012] Furthermore, in Japanese Patent No. 6518800 (Patent Document 7), a method for producing a functional material containing plasmalogen has been proposed, which can efficiently extract plasmalogen from animal tissues and obtain a functional material containing a large amount of plasmalogen

[0013] This method for producing a functional material containing plasmalogen includes an enzyme treatment step of treating tissues of animals selected from scallops and sea squirts with a neutral protease and an extraction step of extracting the animal tissues treated with the neutral protease with an extract containing ethanol (however, excluding those treated with a lipolytic enzyme simultaneously with or after treatment with a neutral protease).

Prior Art Documents

Patent Documents

[0014]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

Patent Document 6

Patent Document 7

Non-Patent Documents

[0015]

Non-Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0016] In the extraction method disclosed in Patent Document 5, in the extraction step, it is necessary to add a mixed solvent of n-hexane / ethanol / water in an amount about 3 to 5 times the volume of the dried product of the aquatic animal. Therefore, further improvement is required in the amount of solvent used during extraction. Furthermore, in the extraction step, it is necessary to partition the hexane layer and the aqueous layer under predetermined conditions and recover only the hexane layer. Therefore, the treatment process is complicated, and further improvement is required in the extraction step itself, including the partitioning conditions and the partitioning method.

[0017] The production method disclosed in Patent Document 6 specifically hydrolyzes diacyl-type glycerophospholipids by using phospholipase A1 (PLA1) as an enzyme so that only ether-type glycerophospholipids are easily extracted. That is, it utilizes the property that PLA1 cannot decompose ether-type glycerophospholipids. Therefore, in the production method disclosed in Patent Document 6, after drying the primary extract from animal tissue as a raw material (extracted from biological materials or their tissues before enzyme treatment), it is necessary to resuspend it in an enzyme solution and perform an enzyme reaction. Therefore, in order to perform a more efficient enzyme reaction, it is necessary to form an emulsion. However, since the enzyme solution is aqueous with respect to the lipid extract, there is a problem that emulsion formation is not easy. Furthermore, if the amount is small, emulsion formation is possible by using ultrasonic treatment or the like, but it becomes a major issue during scale-up.

[0018] In the manufacturing method disclosed in Patent Document 7, while the raw material animal tissue contains about 70% water, ethanol has a hydroxyl group (-OH). Thus, the hydroxyl group and water molecules are hydrated by hydrogen bonding. As a result, a large amount of ethanol had to be used for the raw materials during extraction. Furthermore, in a general environment, flammable dangerous substances such as ethanols are restricted in their usage amounts, so there was also a problem in that the amount that could be used in one extraction was restricted.

[0019] Therefore, there is a demand for a method that reduces the amount of solvent used during extraction, is easy to scale up, and manufactures ether-type glycerophospholipid-containing functional materials more simply and efficiently.

[0020] In view of this situation, the present invention has been earnestly studied with the aim of providing a method that reduces the amount of solvent used during extraction, is easy to scale up, and efficiently manufactures a large amount of a functional material containing a large amount of ether-type glycerophospholipid.

[0021] As a result, it was found that by selecting a dried biological material, particularly bivalve tissue, as the raw material and subjecting it to an extraction treatment using an ethanol / hexane mixture as the extraction solvent, the amount of solvent used during extraction is reduced and it becomes easy to scale up, and a large amount of an ether-type glycerophospholipid-containing functional material can be manufactured more simply and efficiently, thus completing the present invention.

Means for Solving the Problems

[0022] That is, the invention according to claim 1 of the present invention includes performing an extraction treatment on a biological material using an ethanol / hexane mixture, the biological material is Scallop tissue dried and the water content of the biological material is 2% or less. The mixing ratio of the ethanol and the hexane is, by volume ratio, ethanol:hexane = 1:1~2:3 and the total usage amount of the ethanol / hexane mixture is 100 to 500 mL with respect to 100 g (dry mass) of the biological material and is a method for producing a functional material containing ether-type glycerophospholipid, characterized thereby.

Advantages of the Invention

[0023] The method for producing an ether-type glycerophospholipid-containing composition of this invention includes performing an extraction treatment on a biological material using an ethanol / hexane mixture, and selects a dried biological material as the biological material. Therefore, according to this production method, the amount of the solvent used during extraction is reduced, scale-up is easy, and it becomes possible to efficiently produce a large amount of a functional material containing a large amount of ether-type glycerophospholipid. In addition, in this invention, The biological material is dried scallop tissue, the water content of the biological material is 2% or less, the mixing ratio of the ethanol and the hexane is, by volume ratio, ethanol:hexane = 1:1~2:3 and the total usage amount of the ethanol / hexane mixture is 100 to 500 mL with respect to 100 g (dry mass) of the biological material.

[0024] In the production method, as the biological material, a bivalve tissue, preferably a scallop tissue, can be selected. Since the scallop tissue contains ether-type phospholipids containing a large amount of long-chain polyunsaturated fatty acids such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), with such a configuration, it becomes possible to obtain a large amount of a functional material having more excellent functions.

[0025] Furthermore, in the manufacturing method, the mixing ratio of ethanol and hexane in the ethanol / hexane mixture can be ethanol:hexane = 1:0.1 to 1:10, more preferably 3:1 to 1:3, and even more preferably 3:2 to 1:3, by volume ratio. With such a configuration, the amount of the solvent used during extraction can be further reduced, which is economically more advantageous.

[0026] Furthermore, in the manufacturing method, the water content of the biological material can be adjusted to 25% or less, more preferably 15% or less, and even more preferably 2% or less. With such a configuration, the amount of the solvent used during extraction can be further reduced, which is economically even more advantageous.

Brief Description of the Drawings

[0027]

Figure 1

Embodiments for Carrying Out the Invention

[0028] Hereinafter, embodiments of the manufacturing method of the ether-type glycerophospholipid-containing functional material according to the present invention will be described. It should be noted that the present invention will be described mainly with reference to preferred representative examples, but the present invention is not limited to such representative examples.

[0029] The manufacturing method of the ether-type glycerophospholipid-containing functional material of the present invention includes performing an extraction process on a biological material using an ethanol / hexane mixture. The manufacturing method with such a configuration can reduce the amount of the solvent used during the extraction of the ether-type glycerophospholipid-containing functional material, which is economically advantageous and enables obtaining a large amount of the ether-type glycerophospholipid-containing functional material efficiently. As a result, according to the manufacturing method, it becomes possible to contribute to the commercial production of functional materials containing ether-type glycerophospholipids.

[0030] The biological material includes those containing the ether-type glycerophospholipid, and may be dried and is not particularly limited.

[0031] Examples of the biological material include animals, plants, and microorganisms. As the biological material, animals or their tissues are preferably selected because they have a higher content of ether-type glycerophospholipids compared to plant tissues and microorganisms and are easily available in large quantities at low cost. Examples of the animals include mammals, birds, and fish and shellfish.

[0032] As the mammals, livestock are suitable from both the aspects of supply stability and safety. Examples include mammals such as cows, pigs, horses, goats, sheep, deer, camels, llamas, and poultry such as chickens, ducks, turkeys, and ostriches. In the case of the mammals, the main tissues containing ether-type glycerophospholipids include skin, brain, intestine, heart, and genital organs.

[0033] As the fish and shellfish, those that can be bred, that is, farmed, are suitable. 1) Fishes such as amberjack, red sea bream, rainbow trout, yellowtail, flounder, pufferfish, striped jack, Spanish mackerel, greater amberjack, Japanese sea bass, sea bass, cedar, bluefin tuna, kuruma shrimp, carp, eel, rainbow trout, ayu, yamame, amago, Japanese dace, Amur dace, and Yamato dace 2) Crustaceans such as kuruma shrimp, black tiger shrimp, tiger prawn, and horseshoe crab 3) Mollusks such as abalone, turban shell, scallop, and oyster are exemplified. Among them, mollusks such as abalone, turban shell, scallop, and oyster, particularly bivalves, are more suitable. In the case of the seafood, the main tissues containing ether-type glycerophospholipids include viscera, gonads, muscles and the like.

[0034] In the present invention, from the viewpoints that the content rate of neutral lipids in total lipids is low, the content rate of phospholipids is high, and further the content rate of ether-type glycerophospholipids (particularly those containing a large amount of long-chain polyunsaturated fatty acids such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA)) in phospholipids is high, that the long eating experience and the large amount of landed catch are prominent, and that there are cases where tissues other than the main edible part, scallop adductor muscle, are discarded, it is more preferable to select scallops. Here, in the present invention, scallops refer to edible bivalves belonging to the family Pectinidae. Examples of the scallops include those belonging to the genus Mizuhopecten and the genus Pecten. can be mentioned. More specifically, as the scallops, scallops (Mizuhopecten yessoensis) landed in Japan, European scallops (Pecten maximus) collected in Europe, and the like can be mentioned. In addition, examples of the tissues of the scallops include scallop adductor muscle, mantle, gonad and the like.

[0035] As the microorganism, for example, bacteria belonging to the genus Propionibacterium can be used. In the case of bacteria, the "tissue" is the bacteria themselves.

[0036] In the production method of the present invention, as the biological material, a dried one is selected. With such a configuration, since the raw material has a low water content, it is possible to reduce the amount of solvent during extraction, and even with a single extraction treatment, it is possible to obtain a functional material containing a large amount of ether-type glycerophospholipids. Regarding the drying, it can be carried out by known methods such as sun drying, hot air drying, cold air drying, vacuum drying, freeze drying, etc. From the perspective of not destroying the active ingredients in the functional material, freeze drying is preferably selected.

[0037] Regarding the drying conditions, there are no particular restrictions, and they can be appropriately selected according to the biological material selected as the raw material. Preferably, the drying is carried out so that the moisture content of the biological material is 25% or less, more preferably 15% or less, and even more preferably 2% or less. With such a configuration, it is possible to further reduce the amount of the solvent used during extraction, which is economically advantageous.

[0038] Regarding the biological material and its tissue, in order to improve the efficiency of the extraction process, before the extraction process, treatments such as cutting, slicing, mincing, and pulverization may be performed in advance. Furthermore, regarding the biological material and its tissue, in order to improve the efficiency of the extraction process, during the extraction process, treatments such as a high-speed homogenizer or a homomixer treatment may be performed.

[0039] In this invention, as the solvent used during the extraction process, that is, the extraction solvent, an ethanol / hexane mixture is selected.

[0040] In the ethanol / hexane mixture, there are no particular restrictions on the mixing ratio of ethanol and hexane. However, the mixing ratio of ethanol and hexane is by volume ratio, preferably ethanol:hexane = 1:0.1 to 1:10, more preferably 3:1 to 1:3, even more preferably 1:0.5 to 1:3, and even more preferably 1:1 to 1:3. For example, an ethanol / hexane (2 / 3 (volume ratio)) mixture can be used. With such a configuration, it is possible to significantly reduce the amount of the solvent used during extraction.

[0041] Regarding the extraction process, it can be carried out by known methods such as the immersion method (cold immersion, warm immersion, etc.), the vacuum immersion method, the pressure immersion method, etc. Also, regarding the conditions of the extraction process such as the extraction temperature and extraction time, they may be appropriately selected according to the biological material selected as the raw material, etc., and there are no particular restrictions. For example, when dried scallop tissue is selected as the raw material, the extraction process can be carried out by stirring at a temperature of 25 °C for 60 minutes or more. In this invention, regarding the number of times of the extraction process, once is sufficient, but it may also be carried out a plurality of times.

[0042] Regarding the amount of the solvent used in one extraction during the extraction process, it may be appropriately selected according to the biological material selected and the extraction method, etc., and there are no particular restrictions. However, in this invention, the amount of the solvent is reduced compared to the prior art. Here, the reduced amount of the extraction solvent compared to the prior art means that for 100 g (dry mass) of the dried biological material (for example, dried scallop tissue) as the raw material, the total amount of the solvent used is about 100 to 3000 mL, preferably about 200 to 2000 mL, more preferably about 250 to 1000 mL.

[0043] In the manufacturing method of this invention, for example, after the extraction process, as an operation to separate the solid content (raw material) and the extract, filtration, etc. can be carried out. If necessary, even if a step of drying and solidifying the separated and recovered extract or a step of concentrating the extract to increase the concentration of the ether-type glycerophospholipid contained therein is added, as long as the object and effect of this invention are not inhibited, the additional steps can be included in this invention together. . Note that for the filtration, for example, filter separation using a metal filter such as filter paper or stainless steel, or centrifugation can be adopted.

[0044] For example, after performing the extraction process, the obtained solid content is removed by filtration or the like to recover the extract, and if necessary, it is dried and solidified to produce an ether-type glycerophospholipid-containing functional material.

[0045] In the ether-type glycerophospholipid-containing functional material obtained by such a production method, a large amount of ether-type glycerophospholipid is contained. Specifically, the amount of ether-type glycerophospholipid contained in the functional material is usually 10 to 500 mg / g, preferably 100 to 500 mg / g.

[0046] In this invention, since the ether-type glycerophospholipid-containing functional material contains ether-type glycerophospholipid as an active ingredient, when it is ingested into the living body, the action and effect of this ether-type glycerophospholipid can be exhibited. Therefore, the ether-type glycerophospholipid-containing functional material can be effectively used as an ether-type glycerophospholipid-containing composition, and such an ether-type glycerophospholipid-containing composition is also included in the technical scope of this invention.

[0047] Specifically, the ether-type glycerophospholipid-containing functional material is extremely effective in the prevention, treatment, and improvement of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, depression, schizophrenia, metabolic syndromes such as diabetes, lipid abnormalities, insomnia, various infectious diseases and immune abnormalities, atopic dermatitis, fatigue, liver dysfunction, and decline of motor muscles.

[0048] The ether-type glycerophospholipid-containing functional material obtained by such a production method can be used as it is in a liquid state, but if desired, it can also be used in various forms such as slurry, semi-solid, and solid by appropriately using known treatment methods. For example, when a liquid is desired as the product form of the functional material, it can be concentrated by vacuum concentration, reverse osmosis membrane concentration, etc., and when a solid is desired, it can also be made into a powder by spray drying, freeze drying, etc.

[0049] Excipients such as cyclic oligosaccharides, dextrin, and starches may be added to the ether-type glycerophospholipid-containing functional material obtained by the production method of the present invention. When the excipient is added, the ether-type glycerophospholipid-containing functional material becomes easy to handle.

[0050] The addition amount of the excipient is preferably 50 to 2000% by mass based on the content of the ether-type glycerophospholipid.

[0051] The ether-type glycerophospholipid-containing functional material can be used as a material for food and drink, or as a raw material for pharmaceuticals, cosmetics, quasi-drugs, and designated quasi-drugs. Such food and drink, pharmaceuticals, cosmetics, quasi-drugs, and designated quasi-drugs may be produced according to known methods.

[0052] Furthermore, for the ether-type glycerophospholipid-containing functional material, various forms of food and drink that are known or will be developed in the future can be appropriately adopted as described above. In this case, the form of functional food or food for specified health use can also be adopted in the same manner.

[0053] As various forms of food and drink products, for example, 1) Beverages such as soft drinks, green tea beverages, black tea beverages, coffee beverages, fermented tea beverages (such as oolong tea), vegetable juices, milk, milk beverages, fermented milk beverages, drink agents, sports beverages, jelly beverages, alcoholic beverages 2) General foods such as jelly-like foods, frozen desserts, cakes, candies, caramels, chewing gums, Japanese confectioneries, snack foods, chocolates, ramune candies, gummies, puddings, yogurts, soups, miso soups, rice, rice balls, meat or processed meat, bread, udon, soba, ramen, pasta, konjac, pickles, natto, tempura flour, wheat flour, arrowroot powder, gelatin, breadcrumbs, kneaded products, retort foods, frozen foods, chilled foods, instant foods 3) Toppings, sauces, soy sauce, fish sauce, miso, cooking sake, vinegar, mirin, oyster sauce, gravy, mayonnaise, ketchup, salt, spices, herbs, curry powder, cooking oil, noodle soup stock, umami seasonings, spices, flavor seasonings, and other seasonings 4) Processed foods such as capsules, tablets, sugar-coated tablets, granules, powders, liquids, edible film agents, jelly agents, etc. 5) Supplements Various products such as these can be listed.

[0054] As forms of supplement products, for example, various products such as soft gels, tablets, powders, etc. can be listed. In particular, when soft gels and powders are selected, it is also possible to provide a state in which a predetermined amount of the ether-type glycerophospholipid-containing functional material is contained in capsules.

[0055] In addition, the food and drink may contain well-known additives that can be contained in foods (for example, functional foods) as necessary within a range that does not impair the properties of the ether-type glycerophospholipid, which is the active ingredient.

[0056] Examples of the additives include water, saccharides, sugar alcohols, starches and modified starches, dietary fiber, milk, processed milk, soy milk, fruit juice, vegetable juice, fruits and vegetables and their processed products, proteins, peptides, amino acids, extracts of animal and plant crude drugs, naturally derived polymers (such as collagen, hyaluronic acid, chondroitin, etc.), vitamins, minerals, thickeners, emulsifiers, preservatives, coloring agents, fragrances, etc. can be listed.

[0057] When the ether-type glycerophospholipid-containing functional material is used as a raw material for pharmaceuticals, pharmaceutically acceptable bases, carriers, additives (such as excipients, binders, disintegrants, lubricants, solvents, sweeteners, coloring agents, flavoring agents, odor-masking agents, surfactants, humectants, preservatives, pH adjusters, thickening agents, etc.) can be formulated as necessary within a range that does not impair the properties of the ether-type glycerophospholipid, which is the active ingredient. Such base materials, carriers, additives, etc. are specifically described in, for example, the Pharmaceutical Additive Dictionary 2000 (Yakujutsu Nippo Co., Ltd.), so, for example, those described therein can be used.

[0058] Also, the dosage form is not particularly limited, and the active ingredient and other ingredients are mixed by a conventional method and can be prepared into preparations such as tablets, coated tablets, powders, granules, fine granules, capsules, pills, liquids, suspensions, emulsions, jelly agents, chewable agents, soft tablets, etc.

[0059] When using the ether-type glycerophospholipid-containing functional material as a raw material for cosmetics, examples of the product form of cosmetics include various products such as emulsions, creams, lotions, oils, packs, facial washes, cleansers, shampoos, rinses, conditioners, soaps, body washes, etc. Cosmetics containing the ether-type glycerophospholipid-containing functional material are effective in improving and preventing skin aging.

[0060] When using the ether-type glycerophospholipid-containing functional material as a raw material for quasi-drugs, examples of the product form of quasi-drugs include those conforming to Items 1 to 3 of Paragraph 2, Article 2 of the Pharmaceutical Affairs Law, such as pharmaceutical cosmetics, hair dyes, and toothbrushing for preventing periodontal disease and dental caries, and those designated by the Minister of Health, Labour and Welfare.

[0061] When using the ether-type glycerophospholipid-containing functional material as a raw material for designated quasi-drugs, examples of the product form of designated quasi-drugs include those conforming to Item 1 of Paragraph 2, Article 2 of the Pharmaceutical Affairs Law, such as vitamin agents, throat lozenges, and stomachic lozenges, and newly designated quasi-drugs and newly defined quasi-drugs designated by the Minister of Health, Labour and Welfare.

[0062] In addition, although it is convenient to use the ether-type glycerophospholipid-containing functional material by mixing it with the product to be used, it goes without saying that the product contains an effective amount of ether-type glycerophospholipid to exhibit the expected effects.

[0063] In the ether-type glycerophospholipid-containing functional material, the intake or dosage of the ether-type glycerophospholipid as the active ingredient varies depending on the age, body weight, constitution, physical condition, dosage form of the drug, administration method, intake or administration period, etc. of the subject. In this case, for example, in the case of oral administration, this can be selected with a dosage of generally 0.05 to 50 mg, more preferably 0.1 to 10 mg per day per adult (body weight about 60 kg) as a guide. In addition, it may be ingested once or divided into multiple times (preferably 2 to 3 times) a day.

Examples

[0064] Hereinafter, examples will be given to explain in detail the manufacturing method of the ether-type glycerophospholipid-containing functional material of the present invention. Note that the present invention is not limited by these examples.

[0065] [Examples 1 to 6] (Manufacture of Ether-Type Glycerophospholipid-Containing Functional Material Derived from Scallop) Using various raw material materials and extraction solvents shown in Table 1 below, various ether-type glycerophospholipid-containing functional materials were manufactured by the following manufacturing method. Note that the amount of the obtained extract was shown as the amount (g) obtained per 100 g of the raw material material used. The results are shown in Table 3.

[0066] <Manufacturing Method> To 50 g of the powder of dried scallop shell tissue (mantle part), 250 mL of an ethanol / hexane mixed solution was added, and the mixture was stirred at room temperature for 1 hour. Thereafter, the obtained mixed solution was subjected to suction filtration treatment, and the liquid layer was recovered. The solvent was distilled off from the recovered liquid layer using a rotary evaporator to obtain a dried solid.

[0067]

Table 1

[0068] [Comparative Example 1] (Production of Functional Material Containing Ether-Type Glycerophospholipid Derived from Scallop) A functional material containing an ether-type glycerophospholipid was produced by the following production method. The amount of the obtained extract was shown in terms of the amount (g) obtained per 100 g of the raw material used. The results are shown in Table 3.

[0069] [Production Method] To 200 g of fresh raw scallop tissue (mantle part), 300 mL of a hexane / ethanol (volume ratio 3 / 2) mixed solution was added and pulverized with a blender. To the obtained mixed solution, an additional 700 mL of a hexane / ethanol (volume ratio 3 / 2) mixed solution was added and stirred at room temperature for 1 hour. Thereafter, the obtained mixed solution was subjected to suction filtration treatment, and the liquid layer was recovered. The residue was further washed with 400 mL of a hexane / ethanol (volume ratio 3 / 2) mixed solution, and the liquid layer was similarly recovered and combined with the previously obtained liquid layer. To the recovered liquid layer, 800 mL of a 1 g / 15 mL sodium sulfate solution and 30 mL of acetic acid were added, partitioned with a separatory funnel, and the hexane layer was recovered. To the aqueous layer, an additional 300 mL of a hexane / ethanol (volume ratio 3 / 2) mixed solution and 300 mL of saturated brine were added and partitioned again, and the hexane layer was similarly recovered and combined with the previously obtained hexane layer. The solvent was distilled off from the obtained hexane layer using a rotary evaporator to obtain a dried solid. To the obtained dried solid, 40 mL of a phospholipase A1 solution (20 mg / mL, 0.1 M citrate buffer, pH 4) was added and reacted at 50 °C for 1.5 hours. To the obtained reaction product, 360 mL of a hexane / ethanol (volume ratio 3 / 2) mixed solution, 220 mL of water, 7.2 mL of acetic acid, and 40 mL of saturated brine were added, partitioned with a separatory funnel, and the hexane layer was recovered. The solvent was distilled off from the obtained hexane layer using a rotary evaporator to obtain a dried solid.

[0070] [Comparative Example 2] (Production of Ether-Type Glycerophospholipid-Containing Functional Material from Scallop) An ether-type glycerophospholipid-containing functional material was produced by the following production method. The amount of the obtained extract was indicated by the amount (g) obtained per 100 g of the raw material used. The results are shown in Table 3.

[0071] [Production Method] To 50 g of powder of dried scallop tissue (mantle part) (moisture content 1.7%), 250 mL of hexane was added, and the mixture was stirred at room temperature for 1 hour. Thereafter, the obtained mixture was subjected to suction filtration treatment, and the liquid layer was recovered. The solvent was distilled off from the recovered liquid layer using a rotary evaporator to obtain a dried solid.

[0072] [Comparative Example 3] (Production of Ether-Type Glycerophospholipid-Containing Functional Material from Scallop) An ether-type glycerophospholipid-containing functional material was produced by the following production method. The amount of the obtained extract was indicated by the amount (g) obtained per 100 g of the raw material used. The results are shown in Table 3.

[0073] [Production Method] To 50 g of powder of dried scallop tissue (mantle part) (moisture content 1.7%), 250 mL of an isopropanol / hexane mixed solution (volume ratio 2 / 3) was added, and the mixture was stirred at room temperature for 1 hour. Thereafter, the obtained mixture was subjected to suction filtration treatment, and the liquid layer was recovered. The solvent was distilled off from the recovered liquid layer using a rotary evaporator to obtain a dried solid.

[0074] [Comparative Examples 4 and 5] (Production of Functional Material Containing Ether-Type Glycerophospholipid Derived from Scallop) A functional material containing ether-type glycerophospholipid was produced in the same manner as in Example 1, except that the raw material and extraction solvent used in Example 1 were replaced with those shown in the raw material and extraction solvent columns of the sheet material in Table 2 below, respectively.

[0075] [Table 2]

[0076] [Table 3]

[0077] [Results] From Table 3, the amounts of the extracts obtained in Examples 1 to 6 above were all larger than those obtained in Comparative Examples 1 to 5. From the above, it is clear that when producing a functional material containing ether-type glycerophospholipid, it is effective to perform an extraction treatment on the dried biological material using an ethanol / hexane mixture.

[0078] [Evaluation Example] (HPLC Analysis of Functional Material Containing Ether-Type Glycerophospholipid Derived from Scallop) HPLC analysis was performed under the following conditions on 2 mg of the extract (dry matter) obtained in any of Examples 1 to 6 or any of Comparative Examples 2 to 5, or 5 mg of the extract (dry matter) obtained in Comparative Example 1, which was dissolved in 1 mL of a hexane / 2-propanol (3:2) mixture. The amount of the contained ether-type glycerophospholipid was indicated by the amount (g) contained per 100 g of the raw material used. Regarding the increase rate, it was calculated as the ratio of the amount of ether-type glycerophospholipid contained in the extract obtained in Comparative Example 1 measured by HPLC analysis to the amount of ether-type glycerophospholipid contained in the extract (the extract obtained in Examples or Comparative Examples other than Comparative Example 1), with the amount of ether-type glycerophospholipid in Comparative Example 1 being set to 1. The results are shown in Table 4.

[0079] <HPLC Conditions> 1) Equipment used: Shimadzu LC-AD20 system (manufactured by Shimadzu Corporation) 2) Column: LiChrospher Diol 100 (5μm, 250-4, manufactured by Merck Millipore) 3) Flow rate: 1.0 mL / min 4) Column temperature: 50°C 5) Detector: ELSD-LTII (Evaporative Light Scattering Detector) (manufactured by Shimadzu Corporation) 6) Drift tube temperature: 50°C 7) Mobile phase: (A) Hexane / 2-propanol / acetic acid (82:17:1, v / v) + 0.08% trimethylamine) (B) 2-propanol / water / acetic acid (85:14:1, v / v) + 0.08% triethylamine) 8) Gradient: (B) 5%, 0 min → (B) 45%, 12 min → (B) 45%, 21 min → (B) 85%, 22 min → (B) 85%, 27 min → (B) 5%, 27 min → (B) 5%, 37 min

[0080]

Table 4

[0081] <Results> From Table 4, the amounts of ether-type glycerophospholipid obtained in Examples 1 to 6 above were all 4 times or more the amount of ether-type glycerophospholipid obtained in Comparative Example 1. This effect was remarkable when the water content of the biological material was 1.7%. From the above, according to the method for producing an ether-type glycerophospholipid-containing functional material according to the present invention, it is clear that the amount of ether-type glycerophospholipid obtained by a single extraction treatment increases dramatically, which is due to performing an extraction treatment on the dried biological material using an ethanol / hexane mixed solution.

[0082] Figure 1 is a graph showing the relationship between the concentration of ethanol used and the amount of ether-type glycerophospholipid obtained. From Figure 1, when the concentration of ethanol in the ethanol / hexane mixed solution is 25 to 75% in the case of using the ethanol / hexane mixed solution as the extraction solvent during the extraction treatment, it was found that the amount of ether-type glycerophospholipid obtained is 1.4 to 1.8 times that when using ethanol alone (ethanol concentration 100%). Therefore, in the method for producing an ether-type glycerophospholipid-containing functional material according to the present invention, since an ethanol / hexane mixed solution is selected as the extraction solvent during the extraction treatment, a large amount of ether-type glycerophospholipid can be efficiently obtained. By setting the concentration of ethanol contained in this mixed solution to 25 to 75%, that is, setting the mixing ratio to 1 / 3 to 3 / 1 by volume ratio, it is clear that a larger amount of ether-type glycerophospholipid can be obtained more efficiently. In addition, the amount of ether-type glycerophospholipid obtained in the above Comparative Example 3 (when using an isopropanol / hexane mixed solution as the extraction solvent) was more than the amount of ether-type glycerophospholipid obtained in Comparative Example 1, but less than the amount of ether-type glycerophospholipid obtained in Examples 1 to 6.

[0083]

Table 5

[0084] Table 5 shows the amount (usage amount) of the extraction solvent used for 100 g of the raw material (biological material). In Table 5, the reduction rate was calculated as the ratio of the total amount of extraction solvent used in Example 5 to the total amount of extraction solvent used in Comparative Example 1. From Table 5, in this invention, since a dried biological material is selected as the raw material and an ethanol / hexane mixture is selected as the extraction solvent for the extraction treatment, it is clear that the amount of extraction solvent used can be significantly reduced.

[0085]

Table 6

[0086] Furthermore, Table 6 shows the amount of extraction solvent used per 1 g of the obtained ether-type glycerophospholipid. In Table 6, the values in parentheses indicate the ratio of the amount of extraction solvent used in each Example or Comparative Example 4 or 5 to the amount of extraction solvent used in Comparative Example 1. From the results of Example 5 and Comparative Example 1 in Table 6, it was found that according to the method for producing an ether-type glycerophospholipid-containing functional material according to this invention, the amount of extraction solvent used can be reduced to about 1 / 13 at most to obtain the same amount of ether-type glycerophospholipid. Therefore, it is clear that according to the method for producing an ether-type glycerophospholipid-containing functional material according to this invention, the amount of extraction solvent used per unit extraction amount can be reduced by about 90% or more.

Industrial Applicability

[0087] According to this invention, when producing a functional material containing a large amount of ether-type glycerophospholipid, the amount of solvent used in extraction is reduced, scale-up is easy, and it becomes possible to efficiently produce a large amount of such a functional material. Therefore, it is widely used in the industry that handles ether-type glycerophospholipids.

Claims

【Claim 1】 comprising performing an extraction treatment on a biological material using an ethanol / hexane mixture, wherein the biological material is a dried scallop tissue, the moisture content of the biological material is 2% or less, the mixing ratio of the ethanol and the hexane is, by volume ratio, ethanol:hexane = 1:1 to 2:3, the total usage amount of the ethanol / hexane mixture is 100 to 500 mL with respect to 100 g (dry mass) of the biological material and being a method for producing an ether-type glycerophospholipid-containing functional material.

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