WO2017150679A1 - Procédé de séparation de sélénonéïne - Google Patents

Procédé de séparation de sélénonéïne Download PDF

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WO2017150679A1
WO2017150679A1 PCT/JP2017/008326 JP2017008326W WO2017150679A1 WO 2017150679 A1 WO2017150679 A1 WO 2017150679A1 JP 2017008326 W JP2017008326 W JP 2017008326W WO 2017150679 A1 WO2017150679 A1 WO 2017150679A1
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selenonein
reverse osmosis
osmosis membrane
membrane treatment
raw material
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PCT/JP2017/008326
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English (en)
Japanese (ja)
Inventor
倫明 山下
由美子 山下
賢司 石原
伸太朗 今村
卓也 世古
智広 上田
三十郎 藤原
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国立研究開発法人水産研究・教育機構
岩手県
有限会社バイオケム
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Priority to JP2018503406A priority Critical patent/JP6884345B2/ja
Publication of WO2017150679A1 publication Critical patent/WO2017150679A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/56—Materials from animals other than mammals
    • A61K35/60—Fish, e.g. seahorses; Fish eggs
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/66—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives

Definitions

  • the present invention relates to a method for separating selenonein.
  • Serenonein a powerful antioxidant, is thought to contribute to the prevention of lifestyle diseases such as cancer, heart disease, immune disease, type 2 diabetes, and aging, and is useful for functional foods, cosmetics, feeds, reagents, etc. Application is expected.
  • Selenonein is abundant in tissues of squids, fish, birds, mammals, etc., and a method using an organic solvent such as methanol or acetonitrile has been reported as a method for separating and extracting selenonein from the tissues.
  • organic solvent such as methanol or acetonitrile
  • patent document 1 etc. From a viewpoint of the safety improvement at the time of ingestion and use to a living body as a functional food, cosmetics, etc., and a viewpoint of manufacturing cost reduction, water and a food additive are used without using an organic solvent. There has been a demand for a method for efficiently separating and purifying selenonein.
  • the present invention has been made in view of the above-mentioned circumstances, and a problem to be solved is to provide a method for efficiently separating selenonein from a raw material containing selenonein without using an organic solvent. .
  • the present inventors have efficiently used a water-based solvent extract of a raw material material containing selenonein without using an organic solvent by using a reverse osmosis membrane treatment technique. It has been found that selenonein can be separated, and the present invention has been completed by further research based on this finding. That is, the present invention is as follows.
  • a method for separating selenonein comprising subjecting an aqueous solvent extract of a raw material containing selenonein to a reverse osmosis membrane treatment.
  • the method according to [1] comprising subjecting an aqueous solvent extract of a raw material containing selenonein to a reverse osmosis membrane treatment to collect a permeate fraction containing a monomer.
  • the method according to [2], comprising subjecting the permeated fraction to a reverse osmosis membrane treatment to collect a non-permeate fraction containing an oxidized dimer formed from a monomer.
  • selenonein can be efficiently separated from a raw material containing selenonein without using an organic solvent. Further, according to the present invention, even if the raw material containing selenonein is a marine processing residue containing methylmercury, which is a harmful element, selenonein can be separated by removing methylmercury.
  • FIG. 2 is a chromatogram by HPLC-ICP-MS analysis of a permeation fraction of a reverse osmosis membrane treatment using an NF membrane in Example 2.
  • the selenonein separation method of the present invention (hereinafter also simply referred to as “the method of the present invention”) is characterized mainly by subjecting an aqueous solvent extract of a raw material containing selenonein to a reverse osmosis membrane treatment.
  • “selenonein” means a compound represented by formula (I) (3- (2-hydroseleno-1H-imidazol-5-yl) -2- (trimethylamino) propanoate), and tautomers thereof. (3- (2-selenoxo-2,3-dihydro-1H-imidazol-4-yl) -2- (trimethylammonio) propanoate) represented by formula (II), and dimers thereof. It is a general term for a compound (3,3 ′-(2,2′-disanediylbis (1H-imidazole-5,2-diyl)) bis (2- (trimethylamino) propanoate)) represented by a certain formula (III).
  • the compound represented by formula (I) is referred to as “selenol-type monomer”
  • the compound represented by formula (II) is referred to as “selenoketone-type monomer”
  • formula (III) represented by formula (III).
  • the resulting compound may be referred to as “selenonein oxidized dimer” or simply “oxidized dimer”.
  • the selenol type monomer and the selenoketone type monomer may be collectively referred to as “selenonein monomer” or simply “monomer”.
  • the raw material used in the method of the present invention is not particularly limited as long as it contains selenonein.
  • fish and shellfish e.g. mackerel such as tuna and sesame mackerel, sawaras, salmon, yellowtail, kinmedai
  • Fish tissues e.g. squid such as squid, etc.
  • birds eg poultry etc.
  • mammals eg livestock, cetaceans etc.
  • other animal tissues eg spleen, liver, pancreas, heart and other internal organs, blood, head Parts, muscles, edible parts, etc.
  • plant tissues microbial tissues; chemically synthesized products; fermented products, etc.
  • fishery processed residues for example, seafood and whales
  • Visceral organs blood, muscle, head, edible portion, middle bone, etc.
  • tuna and bonito such as bluefin tuna, southern bluefin tuna, bigeye, albacore, mackerel, mackerel, yellowtail, and snapper muscles (for example, bloody meat etc. ), Blood, head, internal organs (for example, spleen, liver, pancreas, heart, etc.), squid internal organs; whale meat (for example, red meat of white whales) and the like.
  • the amount of selenonein contained in the raw material is not particularly limited, but is usually 1 ppm or more, preferably 2 ppm or more, more preferably 5 ppm or more based on the raw material.
  • the amount of selenonein contained in the raw material is preferably as large as possible, and the upper limit is not particularly limited, but is usually 100 ppm with respect to the raw material.
  • the amount of selenonein contained in the raw material is determined by a fluorescence method using 2,3-diaminonaphthalene (DAN) or a gel permeation chromatography (GPC) column according to the method described in JP2011-121914A. It is measured by high performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS analysis) used.
  • the raw material used in the method of the present invention may be a dried product.
  • the form of the raw material is not particularly limited, and may be, for example, powder, a refrigerated state, a frozen state, or the like.
  • the “aqueous solvent extract of raw material containing selenonein” used in the method of the present invention refers to an extract of raw material containing selenonein using an aqueous solvent as an extraction solvent.
  • the extract may be prepared using a raw material containing selenonein and an aqueous solvent, and may be prepared by appropriately combining known methods or methods equivalent thereto, and is not particularly limited except that an organic solvent is not used as the extraction solvent.
  • a raw material containing selenonein is homogenized by adding an aqueous solvent (for example, water) of about 10 to 10 times by weight, and then centrifuged or filtered with a filter paper, paper towel, cloth, or the like.
  • an aqueous solvent may be added to the raw material, and then heated under boiling conditions (100 ° C.), a high-pressure steam sterilizer (eg, 121 ° C.), or the like.
  • Examples of the aqueous solvent used for the preparation of the aqueous solvent extract of the raw material containing selenonein include water.
  • Specific examples of water include deionized water, mineral water, ultrapure water, reverse osmosis filtered water, seawater, tap water, carbonated water, and the like.
  • Preparation of the aqueous solvent extract of the raw material containing selenonein is preferably performed using a filter medium.
  • a filter medium By using a filter medium, impurities such as protein, lipid, and heme iron can be efficiently removed.
  • the filter medium include diatomaceous earth, acid clay, activated clay, silica gel, activated carbon and the like, and are effective in removing heme iron, which is a browning substance, and are preferably acidic clay, activated clay, or silica gel. is there.
  • These filter media may be used independently or may use 2 or more types together.
  • the use mode of the filter medium is not particularly limited, but it is usually added to the raw material material when the raw material material homogenized by adding an aqueous solvent (for example, water) is subjected to centrifugation or filtration.
  • the addition amount of the filter medium may be appropriately adjusted according to the type of the filter medium and the amount of the raw material, but is usually 0.1 to 15% by weight, preferably 1 to 10% by weight with respect to the raw material. %.
  • the aqueous solvent extract of the raw material containing selenonein may be added with urease and / or protease before the reverse osmosis membrane treatment.
  • urease By adding urease, urea contained in the aqueous solvent extract can be decomposed and removed.
  • the amount of urease added is usually from 0.001 to 3% by weight, preferably from 0.01 to 1% by weight, more preferably from 0.05 to 0.2% by weight, based on the raw material.
  • the soluble protein contained in the aqueous solvent extract can be decomposed and removed by adding a protease.
  • the amount of protease added is usually from 0.001 to 3% by weight, preferably from 0.01 to 1% by weight, more preferably from 0.1 to 0.5% by weight, based on the raw material.
  • Protease can be inactivated by heating to 80 ° C. or higher after degradation of soluble protein.
  • Protease can be added using, for example, an enzyme preparation containing protease (eg, pancrelipase, pronase, etc.).
  • a phosphatase may be added to the aqueous solvent extract of the raw material containing selenonein before the reverse osmosis membrane treatment.
  • phosphatase By adding phosphatase, it is possible to remove phosphoric acid from the phosphorylated saccharide contained in the aqueous solvent extract and to suppress the formation of a complex with phosphonone in the selenonein, thus increasing the purity of selenonein.
  • the phosphatase used in the present invention is not particularly limited as long as it can remove phosphate from the phosphorylated saccharide, and both acid phosphatase and alkaline phosphatase can be used.
  • the amount of phosphatase added is usually 0.001 to 3% by weight, preferably 0.01 to 1% by weight, more preferably 0.1 to 0.3% by weight, based on the raw material.
  • the phosphatase may be added alone to the aqueous solvent extract, or may be added to the aqueous solvent extract in combination with the above urease and / or protease.
  • the aqueous solvent extract of the raw material containing selenonein may be brought into contact with at least one selected from the group consisting of a cation exchange resin, an anion exchange resin, silica gel and a synthetic adsorption resin before the reverse osmosis membrane treatment. It is preferable to contact all of them. By contacting with at least one of these resins and silica gel, impurities such as heme, salts, pigments and lipids contained in the aqueous solvent extract can be removed.
  • the synthetic adsorption resin is not particularly limited as long as it does not have an ion exchange group, and examples thereof include styrene-divinylbenzene synthetic adsorption resin and acrylic synthetic adsorption resin. preferable. Specific examples of the styrene-divinylbenzene synthetic adsorption resin include Sepabeads HP20 and Sepabeads SP207 (manufactured by Mitsubishi Chemical Corporation).
  • a method of bringing the aqueous solvent extract of the raw material containing selenonein into contact with at least one selected from the group consisting of a cation exchange resin, an anion exchange resin, silica gel, and a synthetic adsorption resin is not particularly limited.
  • a column system in which at least one silica gel is packed in a column and an aqueous solvent extract is passed through the column is preferable.
  • aqueous solvent extract of the raw material containing selenonein is brought into contact with at least two (preferably all) selected from the group consisting of a cation exchange resin, an anion exchange resin, silica gel and a synthetic adsorption resin, for example, At least two (preferably all) of a column packed with a cation exchange resin, a column packed with an anion exchange resin, a column packed with silica gel, and a column packed with a synthetic adsorption resin are connected in series.
  • An aqueous solvent extract may be passed.
  • the order of the columns to be connected is not particularly limited, and can be changed as appropriate.
  • the cation exchange resin and the anion exchange resin may be mixed and packed in the same column. When insoluble fine particles are generated, the fine particles can be removed by passing an aqueous solvent extract through a micropore filter.
  • the cation exchange resin a “strongly acidic cation exchange resin” described later may be used.
  • reverse osmosis membrane treatment means that a solution having different solute concentrations (or a solution and a pure solvent) is partitioned by a semipermeable membrane (reverse osmosis membrane) that permeates the solvent and does not permeate the solute. Apply a pressure greater than the osmotic pressure difference to the (or solution side) and permeate the solvent from the high concentration solution side to the low concentration solution side (or from the solution side to the pure solvent side) to separate the solute from the solution. Refers to processing.
  • the aqueous solvent extract of the raw material containing selenonein usually contains both monomer and oxidized dimer, and the monomer mainly combines with heme, ergothioneine, urea, etc. to form inclusion compounds.
  • the monomer can be dissociated from the inclusion compound, and selenonein can be separated with high purity.
  • selenonein can be efficiently concentrated by reverse osmosis membrane treatment.
  • the reverse osmosis membrane used in the method of the present invention is preferably one that allows permeation of selenonein monomer but low permeability of the oxidized dimer and can concentrate the oxidized dimer in the non-permeate fraction.
  • the molecular weight cutoff of the reverse osmosis membrane used in the method of the present invention is preferably 100 to 1000, more preferably 300 to 500.
  • the NaCl blocking rate of the reverse osmosis membrane used in the method of the present invention is preferably 30 to 99.9%, more preferably 50 to 90%.
  • the pore size of the reverse osmosis membrane used in the method of the present invention is usually 2 nanometers or less.
  • reverse osmosis membranes that can be used in the method of the present invention include filtration membranes for seawater desalination; nanofiltration (NF) membranes mainly used for desalination and food extract production. .
  • An NF film having a low rejection of ions or salts of 70% or less may be used.
  • These reverse osmosis membranes may be used alone or in combination of two or more.
  • the filtration membrane for seawater desalination refers to a reverse osmosis membrane having a NaCl rejection rate of 99% or more.
  • CSM-2540SR manufactured by Wongjin Chemical
  • DOW FILMTEC registered trademark
  • SW30 manufactured by The Dow Chemical Company
  • the NF membrane refers to a reverse osmosis membrane having a molecular weight cut off of 200 to 100.
  • NF 50% Rejection Membrane manufactured by M-N2521A5, Applied Membranes, Inc.
  • NF 90% Rejection Membrane M-N2521A, M-N2521A
  • DOW FILMTEC registered trademark
  • NF270 manufactured by The Dow Chemical Company
  • DOW FILMTEC registered trademark
  • the material of the reverse osmosis membrane used in the method of the present invention is not particularly limited, and examples thereof include cellulose acetate, aromatic polyamide, polybenzimidazolone, polyacrylonitrile, polysulfone, and composite membranes thereof. .
  • the shape of the reverse osmosis membrane used in the method of the present invention is not particularly limited, and examples thereof include a spiral type, a tubular type, a capillary type, a hollow fiber type, and a flat membrane type.
  • the filtration pressure in the reverse osmosis membrane treatment is usually 0.1 to 1.6 MPa, preferably 0.2 to 0.5 MPa.
  • the embodiment of the reverse osmosis membrane treatment in the method of the present invention is not particularly limited, and the type of reverse osmosis membrane, the number of reverse osmosis membrane treatments, the fraction to be collected, etc. can be appropriately selected according to the purpose.
  • the aqueous solvent extract of the raw material material containing selenonein is subjected to the reverse osmosis membrane treatment to collect the permeate fraction from which the monomer has eluted, The permeated fraction can be subjected again to the reverse osmosis membrane treatment to recover the non-permeated fraction enriched in the oxidized dimer.
  • the monomer dissociated from the inclusion compound and eluted into the permeated fraction is rapidly oxidized with air to form an oxidized dimer.
  • the oxidized dimer can be concentrated in the non-permeate fraction.
  • impurities that are low in permeability to the reverse osmosis membrane for example, amino acids, peptides, urea, etc.
  • the permeability to the reverse osmosis membrane can remove both high-impurity impurities (for example, salts, amines, etc.), and high-purity selenonein can be obtained.
  • the aqueous solvent extract of the raw material containing selenonein may be subjected to a reduction treatment using a reducing agent or the like before the reverse osmosis membrane treatment. Since the oxidized dimer dissociates into a monomer when reduced, the reduced amount of the aqueous solvent extract is subjected to a reverse osmosis membrane treatment using a reverse osmosis membrane that permeates the monomer. A permeate fraction containing the above monomer can be obtained.
  • the reducing agent used in the method of the present invention is not particularly limited, and examples thereof include cysteine, cystine, glutathione, sulfite (eg, sodium sulfite), oxalic acid and the like.
  • the oxidized dimer When the oxidized dimer is concentrated in the non-permeating fraction, for example, by using a filtration membrane for seawater desalination, components contained in food extracts and the like (eg, amino acids, peptides, ergothioneine, salt, carbohydrates, nucleic acids) Etc.), or by using an NF membrane, the oxidized dimer can be concentrated while removing salt, ammonium salt, acetic acid, amino acid, urea and the like through the membrane. When the oxidized dimer is concentrated while removing salt and the like through the membrane, the salt and the like can be sufficiently removed by diluting the selenonein solution with purified water before the reverse osmosis membrane treatment.
  • a filtration membrane for seawater desalination components contained in food extracts and the like (eg, amino acids, peptides, ergothioneine, salt, carbohydrates, nucleic acids) Etc.), or by using an NF membrane
  • the oxidized dimer can
  • the embodiment of the reverse osmosis membrane treatment in the method of the present invention is not limited to the above-described embodiment.
  • a permeation in which an aqueous solvent extract of a raw material material containing selenonein is subjected to a reverse osmosis membrane treatment to elute monomers. After collecting the fraction, the permeated fraction is subjected to reverse osmosis membrane treatment, and the permeated fraction is collected again to remove impurities such as heme, ergothioneine, and urea that form inclusion compounds with the monomer. It can be further removed.
  • the aqueous solvent extract of the raw material containing selenonein is subjected to reverse osmosis membrane treatment, and after collecting the non-permeate fraction in which the oxidized dimer is concentrated together with the components contained in the food extract,
  • the non-permeated fraction enriched in the oxidized dimer may be recovered while subjecting the fraction to the reverse osmosis membrane treatment to remove sodium chloride and the like through the membrane.
  • the aqueous solvent extract of the raw material containing selenonein is subjected to a reverse osmosis membrane treatment, and after collecting the permeated fraction from which the monomer was eluted and the non-permeated fraction from which the oxidized dimer was concentrated, The permeation fraction and the non-permeation fraction may be mixed, and the obtained mixture may be subjected to a reverse osmosis membrane treatment again to recover the non-permeation fraction enriched in the oxidized dimer.
  • the monomer eluted in the permeate fraction is rapidly oxidized in the air to form an oxidized dimer. Therefore, the permeate fraction is mixed with the non-permeate fraction enriched in the oxidized dimer.
  • an aqueous solvent extract of a raw material material containing selenonein is subjected to a reverse osmosis membrane treatment (preferably, a reverse osmosis membrane treatment using a filtration membrane and / or an NF membrane for seawater desalination). It is preferable to include a fraction and / or a non-permeate fraction, and the permeate fraction and / or the non-permeate fraction is treated with a reverse osmosis membrane (preferably, a filtration membrane and / or an NF membrane for seawater desalination).
  • a reverse osmosis membrane treatment preferably, a reverse osmosis membrane treatment using a filtration membrane and / or an NF membrane for seawater desalination.
  • an aqueous solvent extract of a raw material containing selenonein is subjected to a reverse osmosis membrane treatment (preferably, a reverse osmosis membrane treatment using a filtration membrane and / or an NF membrane for seawater desalination).
  • a reverse osmosis membrane treatment preferably, a reverse osmosis membrane treatment using a filtration membrane and / or an NF membrane for seawater desalination.
  • a reverse osmosis membrane treatment preferably, a reverse osmosis membrane treatment using a filtration membrane for seawater desalination and / or an NF membrane. It is particularly preferred to include recovering the non-permeate fraction containing the oxidized dimer formed from the monomer.
  • the selenonein separated by the reverse osmosis membrane treatment may be appropriately purified.
  • purification process can be performed combining suitably the known method or the method according to this suitably,
  • the method is not specifically limited, For example, the method etc. which contact selenonein with an ion exchange resin are mentioned.
  • the ion exchange resin for example, a strong acid cation exchange resin, a weak acid cation exchange resin, an anion exchange resin, a reverse phase HPLC column or the like can be used.
  • selenonein can be bound, and high purity selenonein can be used.
  • a strong acid cation exchange resin is preferred because it is effective for production.
  • strongly acidic cation exchange resin refers to a cation exchange resin having a strongly acidic functional group (eg, sulfonic acid group, etc.).
  • Dowex 50W manufactured by Wako Pure Chemical Industries, Ltd.
  • IR120B Olegano Kogyo Co., Ltd.
  • the method for bringing selenonein into contact with an ion exchange resin is not particularly limited, but a solution containing selenonein separated by reverse osmosis membrane treatment is packed in an ion exchange resin (preferably a strong acidic cation exchange resin) in a column. A method of passing liquid is preferred.
  • the equilibration can be performed using 10-50 mM hydrochloric acid or the like, and the adsorbed selenonein is an ammonium salt (eg, ammonium acetate, ammonium formate, chloride). Elution with ammonium).
  • an ammonium salt eg, ammonium acetate, ammonium formate, chloride. Elution with ammonium).
  • Selenonein separated by reverse osmosis membrane treatment may be purified by gel filtration chromatography, reverse phase high performance liquid chromatography or the like.
  • gel filtration chromatography for example, Sephadex G-15, Sepharose S-100 (GE Healthcare), Ultrahydrogel 120 (Waters), etc.
  • the gel filtration column can be equilibrated with 0.1% acetic acid or hydrochloric acid, and may be separated using the solvent.
  • a reverse phase column equilibrated with 0.05 to 10% acetic acid can be used.
  • Selenonein does not adsorb to the reverse phase column and column equilibration and washing can be performed using 0.1-50% acetic acid.
  • the selenonein separated by the reverse osmosis membrane treatment may be further concentrated by, for example, vacuum concentration, ultrafiltration or the like, or may be pulverized by vacuum drying or freeze drying.
  • the selenonein concentrate separated by the method of the present invention preferably has a selenium weight per kg of solids of 0.1 to 1000 mg, more preferably 1 to 500 mg.
  • the selenium weight per 1 kg of the solid is measured by a fluorescence method using DAN or an HPLC-ICP-MS analysis method using a GPC column.
  • the separated selenonein preferably has a selenium content of 1 to 1000 mg / kg when dried, and more preferably 10 to 500 mg / kg.
  • the selenium content during drying is measured by a fluorescence method using DAN or an HPLC-ICP-MS analysis method using a GPC column.
  • the method of the present invention can separate selenonein by removing methylmercury even when the raw material containing selenonein is a fishery processing residue containing methylmercury.
  • the content of methylmercury in the selenonein concentrate separated by the method of the present invention is usually 1 ppm or less, preferably 0.1 ppm or less.
  • the content of methylmercury in the selenonein concentrate is determined according to the method described in Japanese Patent No.
  • a mercury measuring device for example, HG -310 type, manufactured by Hiranuma Sangyo Co., Ltd.
  • selenonein separated by the method of the present invention is obtained without using an organic solvent, for example, food (eg, functional food), nutritional supplement, food additive, feed, feed additive , Pharmaceuticals, veterinary drugs, pharmaceutical additives, cosmetics, cosmetic additives, reagents and the like.
  • a selenonein concentrate can be obtained. Therefore, the method of the present invention can also be referred to as a method for producing a selenonein concentrate.
  • the “selenonein concentrate” refers to a composition containing separated selenonein or the separated selenonein itself.
  • the content of selenonein in the selenonein concentrate is preferably equal to or greater than that of the raw material used, specifically 1 ppm or more, preferably 10 ppm or more.
  • the upper limit of the content of selenonein is not particularly limited, and may be 100% by weight (that is, the selenonein concentrate may be selenonein itself), but is usually 0.1% by weight or less, preferably Is 0.2% by weight or less, more preferably 1% by weight or less.
  • Example 1 Preparation of selenonein concentrate from sesame processing residue After 6.7 kg of sesame processing residue (head, viscera) was crushed with a mincing machine, 32 L of pure water was added and ripened for 5 minutes. After filtering this with a paper towel, 20 ⁇ M L-cystine and 20 ⁇ M sodium sulfite are added as reducing agents, and 2% by weight of acid clay (made by Toshin Kasei Co., Ltd.), 5% by weight of the raw material is used as a filter medium. Of spherical activated carbon (manufactured by Nippon Enviro Chemicals) and 2% by weight of diatomaceous earth were added and stirred for 10 minutes.
  • a filter paper No. 5B
  • a filtrate water extract of processed sesame mackerel residue
  • silica gel 60 manufactured by Nacalai Tesque
  • anion resin manufactured by ASONE
  • synthetic adsorption resin Sepa beads SP207 (manufactured by Mitsubishi Chemical Corporation) are put in filter cartridges (10 inches), and the filtrate 30L is passed through for 6 hours so as to continuously circulate these four types of filters. Deionization treatment was performed.
  • the filtrate after the deionization treatment is diluted by adding 30 L of pure water, and then subjected to a reverse osmosis membrane treatment using a reverse osmosis membrane for seawater desalination (CSM-2540SR, manufactured by Wongjin Chemical Co.). Fractions were collected. As a result, 0.42 mg of selenonein was obtained in terms of selenium. Next, this permeated fraction was subjected to a reverse osmosis membrane treatment using an NF membrane (NF 50% Rejection Membrane, M-N2521A5, Applied Membranes, Inc.), and the non-permeated fraction was collected to remove the permeated fraction. Salt filtration was performed. As a result, 1.7 mg of selenonein was obtained in terms of selenium in this non-permeable fraction.
  • NF membrane NF 50% Rejection Membrane
  • Example 2 Examination of permeability of selenonein to various reverse osmosis membranes After crushing 2kg of bigeye processed residue (blood gourd meat) (selenium content: 7.4mg / kg) with a mincing machine, add 8L of pure water and add 5 Boiled for a minute. To this, 2% by weight of acid clay (made by Toshin Kasei Co., Ltd.), 5% by weight of spherical activated carbon (manufactured by Nippon Enviro Chemicals) and 2% by weight of diatomaceous earth as a filter medium were added and stirred for 10 minutes. Then, it filtered with the filter paper (No. 5B), and obtained the filtrate (water extract of a bigeye processing residue).
  • acid clay made by Toshin Kasei Co., Ltd.
  • spherical activated carbon manufactured by Nippon Enviro Chemicals
  • the filtrate was subjected to reverse osmosis using a reverse osmosis membrane (CSM-2540SR, manufactured by Wongjin Chemical) and NF membrane (NF 50% Rejection Membrane, M-N2521A5, Applied Membranes, Inc.) for producing pure water.
  • CSM-2540SR reverse osmosis membrane
  • NF membrane NF 50% Rejection Membrane, M-N2521A5, Applied Membranes, Inc.
  • FIG. 1 shows a chromatogram by HPLC-ICP-MS analysis of the permeation fraction of reverse osmosis membrane treatment using an NF membrane. Since the peak of selenium near the retention time of 10.3 minutes indicated by the arrow in the figure coincided with the purified selenonein sample, it was confirmed that the selenium compound contained in the permeated fraction was selenonein.
  • a double amount of pure water is added to the filtrate, and a reverse osmosis membrane for seawater desalination (CSM-2540SR, manufactured by Wongjin Chemical) and an NF membrane (NF 50% Rejection Membrane, M-N2521A5, Applied Membranes, Inc.) And the permeated fraction was collected. Furthermore, 60 L of pure water was added to the permeated fraction and subjected to the same reverse osmosis membrane treatment to collect the permeated fraction. The obtained permeation fraction was subjected to a reverse osmosis membrane treatment using an NF membrane (NF 50% Rejection Membrane, M-N2521A5, Applied Membranes, Inc.), and the non-permeation fraction was collected to remove the permeation fraction.
  • CSM-2540SR manufactured by Wongjin Chemical
  • NF membrane NF 50% Rejection Membrane, M-N2521A5, Applied Membranes, Inc.
  • a salt concentration treatment was performed.
  • the obtained non-permeate fraction was dried under reduced pressure using a rotary evaporator, and the dried product containing selenonein was pulverized with a mill.
  • the selenium content of the obtained selenonein powder was 88 mg / kg.
  • a double amount of pure water was added to the filtrate and subjected to a reverse osmosis membrane treatment using a reverse osmosis membrane for seawater desalination (CSM-2540SR, manufactured by Wongjin Chemical Co.) to collect a permeated fraction. Further, 20 L of pure water was added to the permeated fraction, and then subjected to the same reverse osmosis membrane treatment to collect a permeated fraction A and a non-permeable fraction B.
  • CSM-2540SR reverse osmosis membrane for seawater desalination
  • NF membrane NF 50% Rejection Membrane, M-N2521A5, Applied Membranes, Inc.
  • NF 50% Rejection Membrane M-N2521A5, Applied Membranes, Inc.
  • concentration treatment was performed.
  • the obtained non-permeate fraction was dried under reduced pressure using a rotary evaporator, and the dried product containing selenonein was pulverized with a mill.
  • the total amount of selenium in the obtained selenonein powder was 2.2 mg.
  • the non-permeating fraction B contains an oxidized dimer of selenonein, peptides and salts, but a reverse osmosis membrane using an NF membrane (NF 50% Rejection Membrane, M-N2521A5, Applied Membranes, Inc.).
  • a desalting concentration treatment was performed by collecting 2 L of the non-permeate fraction for the treatment.
  • the obtained non-permeate fraction was dried under reduced pressure using a rotary evaporator, and the dried product containing selenonein was pulverized with a mill.
  • the total amount of selenium in the obtained selenonein powder was 5.7 mg.
  • Example 5 Purification of selenonein by ion exchange resin A strongly acidic cation exchange resin column (Dawex 50Wx4, diameter 5 cm x length 20 cm) was equilibrated with 20 mM hydrochloric acid, and selenonein (680 ⁇ g) obtained in Example 1 was obtained. The non-permeate fraction 2L containing was flowed, and selenonein was adsorbed on the column. Next, 1 L of 20 mM hydrochloric acid was flowed to wash the column, and then 0.5 M ammonium chloride was passed through the column to elute selenonein.
  • Example 6 Production of Selenoneine Concentrate from Bluefin Tuna Viscera After 5 kg of frozen bluefin tuna processed residue (including liver and spleen) was crushed with a minced machine, 20 L of activated carbon filtered water was added and ripened at 100 ° C for 5 minutes did. This was filtered with a paper towel (Wipe All X70, manufactured by Nippon Paper Crecia Co., Ltd.) to remove solids, and 20 L of filtrate (water extract of bluefin tuna processing residue) was obtained.
  • a paper towel Wipe All X70, manufactured by Nippon Paper Crecia Co., Ltd.
  • a double amount of pure water was added to the filtrate and subjected to a reverse osmosis membrane treatment using a reverse osmosis membrane (DOW FILMTEC (registered trademark) NF270, manufactured by The Dow Chemical Company). Were collected respectively. Further, this permeate fraction was mixed with the non-permeate fraction, and then 100 mg pronase (manufactured by Roche) was added and subjected to the same reverse osmosis membrane treatment to concentrate the non-permeate fraction. 0.5 kg silica gel was added to the obtained non-permeate fraction and filtered using a filter paper (No. 5A, manufactured by Advantech Toyo Co., Ltd.), and the filtrate was collected.
  • a reverse osmosis membrane DOW FILMTEC (registered trademark) NF270, manufactured by The Dow Chemical Company
  • the filtrate was dried under reduced pressure using a rotary evaporator, and the dried product containing selenonein was pulverized with a mill.
  • the obtained selenonein powder had a selenium concentration of 316 ppm and a total selenium amount of 44 mg.
  • the total mercury concentration of the selenonein powder was 0.218 ppm.
  • the total mercury concentration of the selenonein powder was determined by subjecting the selenonein powder to wet thermal decomposition and then using a mercury measuring apparatus (HG-310, manufactured by Hiranuma Sangyo Co., Ltd.) according to the method described in Japanese Patent No. 5248841. ).
  • Example 7 Production of selenonein concentrate from sesame mackerel processed residue 50 kg of frozen sesame mackerel processed residue (including head, middle bone and internal organs) was crushed with a chopper (punching 9 mm), and then 200 L of clean water was added and 80 Heated at 0 ° C. for 10 minutes. This was filtered with a vibration sieve (100 mesh) to remove solids, and a filtrate was obtained. 10 kg (5% by weight with respect to the extract) of activated carbon, 1 kg (2% by weight with respect to the raw material), acid clay (manufactured by Wako Pure Chemical Industries) and 1 kg (2% by weight with respect to the raw material) ) was added and stirred at 55 ° C. for 20 minutes.
  • this permeate fraction was mixed with the non-permeate fraction, and then 100 mg pronase (manufactured by Roche) was added and subjected to the same reverse osmosis membrane treatment to concentrate the non-permeate fraction.
  • 1 kg of silica gel was added to the obtained non-permeate fraction, followed by filtration using a filter paper (No. 5A, manufactured by Advantech Toyo Co., Ltd.) to remove a brown precipitate. After drying under reduced pressure using a rotary evaporator, the dried product containing selenonein was pulverized with a mill.
  • the selenonein powder obtained had a selenium concentration of 7.1 ppm and a total selenium content of 12 mg.
  • the total mercury concentration of the selenonein powder was 0.125 ppm.
  • Example 8 8 kg of activated carbon filtered water was added to 4 kg of sawara processed residue (residue when three sawaras were lowered. Including head, internal organs, and middle bones), and after crushing finely with a hand mixer, the mixture was heated to boiling. After ripening for 5 minutes, the mixture was filtered through a paper towel (Wipe All X70, manufactured by Nippon Paper Crecia Co., Ltd.) and a colander to remove solids, thereby obtaining 8 L of a filtrate (water extract of Sawara processing residue). To this filtrate, 80 g (2% by weight with respect to the raw material) of diatomaceous earth was added as a filter material and filtered under reduced pressure.
  • sawara processed residue Residue when three sawaras were lowered. Including head, internal organs, and middle bones
  • a selenonein concentrate can be efficiently produced from a raw material containing selenonein without using an organic solvent. Moreover, according to this invention, even if the raw material raw material containing selenonein is a marine processing residue containing methyl mercury which is a harmful element, methylmercury can be removed to produce a selenonein concentrate.

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  • Animal Behavior & Ethology (AREA)
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  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Separation Using Semi-Permeable Membranes (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Treatment Of Water By Ion Exchange (AREA)
  • Water Treatment By Sorption (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

La présente invention concerne un procédé de séparation de manière efficace de sélénonéïne d'un matériau de départ, qui contient de la sélénonéïne, sans utiliser de solvant organique. La présente invention concerne un procédé de séparation de sélénonéïne, qui comprend un procédé dans lequel un extrait de solvant aqueux d'un matériau de départ qui contient de la sélénonéïne est soumis à un traitement sur membrane d'osmose inverse.
PCT/JP2017/008326 2016-03-02 2017-03-02 Procédé de séparation de sélénonéïne WO2017150679A1 (fr)

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WO2023171235A1 (fr) * 2022-03-11 2023-09-14 オルガノ株式会社 Procédé et appareil de production d'eau exempte d'urée, et procédé et appareil d'analyse d'urée
CN118027135A (zh) * 2024-04-11 2024-05-14 黑龙江八一农垦大学 一种从富硒稻米中提取硒多肽的方法

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JP2020125243A (ja) * 2019-02-01 2020-08-20 国立研究開発法人水産研究・教育機構 セレノネインモノマーの分離方法
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WO2023171235A1 (fr) * 2022-03-11 2023-09-14 オルガノ株式会社 Procédé et appareil de production d'eau exempte d'urée, et procédé et appareil d'analyse d'urée
CN118027135A (zh) * 2024-04-11 2024-05-14 黑龙江八一农垦大学 一种从富硒稻米中提取硒多肽的方法

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