JP2012170441A - Fish-derived phospholipid composition and method for producing the same - Google Patents
Fish-derived phospholipid composition and method for producing the same Download PDFInfo
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- JP2012170441A JP2012170441A JP2011038303A JP2011038303A JP2012170441A JP 2012170441 A JP2012170441 A JP 2012170441A JP 2011038303 A JP2011038303 A JP 2011038303A JP 2011038303 A JP2011038303 A JP 2011038303A JP 2012170441 A JP2012170441 A JP 2012170441A
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- phospholipid
- phospholipid composition
- sphingomyelin
- glycerophospholipid
- fatty acid
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- 150000003904 phospholipids Chemical class 0.000 title claims abstract description 177
- 239000000203 mixture Substances 0.000 title claims description 127
- 238000004519 manufacturing process Methods 0.000 title claims description 13
- 150000002327 glycerophospholipids Chemical class 0.000 claims abstract description 66
- 235000014113 dietary fatty acids Nutrition 0.000 claims abstract description 50
- 239000000194 fatty acid Substances 0.000 claims abstract description 50
- 229930195729 fatty acid Natural products 0.000 claims abstract description 50
- 150000004665 fatty acids Chemical class 0.000 claims abstract description 50
- 241000251468 Actinopterygii Species 0.000 claims abstract description 36
- 239000010779 crude oil Substances 0.000 claims abstract description 36
- 239000003921 oil Substances 0.000 claims description 30
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 28
- 239000000470 constituent Substances 0.000 claims description 26
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 claims description 17
- RYCNUMLMNKHWPZ-SNVBAGLBSA-N 1-acetyl-sn-glycero-3-phosphocholine Chemical compound CC(=O)OC[C@@H](O)COP([O-])(=O)OCC[N+](C)(C)C RYCNUMLMNKHWPZ-SNVBAGLBSA-N 0.000 claims description 15
- 125000004432 carbon atom Chemical group C* 0.000 claims description 14
- 150000001982 diacylglycerols Chemical class 0.000 claims description 13
- 241000269851 Sarda sarda Species 0.000 claims description 10
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims 1
- 229910052799 carbon Inorganic materials 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 26
- 239000002994 raw material Substances 0.000 abstract description 10
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- 241000269819 Katsuwonus pelamis Species 0.000 abstract 1
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- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 27
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 26
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 11
- 238000002835 absorbance Methods 0.000 description 9
- 229910052698 phosphorus Inorganic materials 0.000 description 9
- 239000011574 phosphorus Substances 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 7
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 description 7
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 7
- 230000007935 neutral effect Effects 0.000 description 7
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- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
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- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 description 2
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- 235000010469 Glycine max Nutrition 0.000 description 2
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- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
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- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
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- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
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- 150000001298 alcohols Chemical group 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
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- 239000005018 casein Substances 0.000 description 2
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 2
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- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- UKMSUNONTOPOIO-UHFFFAOYSA-N docosanoic acid Chemical compound CCCCCCCCCCCCCCCCCCCCCC(O)=O UKMSUNONTOPOIO-UHFFFAOYSA-N 0.000 description 2
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- 238000002156 mixing Methods 0.000 description 2
- 150000003905 phosphatidylinositols Chemical class 0.000 description 2
- CASUWPDYGGAUQV-UHFFFAOYSA-M potassium;methanol;hydroxide Chemical compound [OH-].[K+].OC CASUWPDYGGAUQV-UHFFFAOYSA-M 0.000 description 2
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- WWUZIQQURGPMPG-UHFFFAOYSA-N (-)-D-erythro-Sphingosine Natural products CCCCCCCCCCCCCC=CC(O)C(N)CO WWUZIQQURGPMPG-UHFFFAOYSA-N 0.000 description 1
- GWHCXVQVJPWHRF-KTKRTIGZSA-N (15Z)-tetracosenoic acid Chemical compound CCCCCCCC\C=C/CCCCCCCCCCCCCC(O)=O GWHCXVQVJPWHRF-KTKRTIGZSA-N 0.000 description 1
- JQWAHKMIYCERGA-UHFFFAOYSA-N (2-nonanoyloxy-3-octadeca-9,12-dienoyloxypropoxy)-[2-(trimethylazaniumyl)ethyl]phosphinate Chemical compound CCCCCCCCC(=O)OC(COP([O-])(=O)CC[N+](C)(C)C)COC(=O)CCCCCCCC=CCC=CCCCCC JQWAHKMIYCERGA-UHFFFAOYSA-N 0.000 description 1
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Abstract
Description
本発明はスフィンゴミエリンを含有するリン脂質組成物及びその製造方法に関する。 The present invention relates to a phospholipid composition containing sphingomyelin and a method for producing the same.
リン脂質は動植物、微生物中に広く分布する、生体膜を構成する主要な成分であり、アルコール類に脂肪酸及びリン酸、塩基などが結合した構造を有する複合脂質である。リン脂質は結合しているアルコールの種類により、グリセロールを含むグリセロリン脂質、スフィンゴシンなどの長鎖アミノアルコールを含むスフィンゴリン脂質に大別される。リン原子の結合様式としてリン酸エステルを含むものとホスホン酸を含むものが区別される場合、天然のリン脂質は４つのグループ、すなわちグリセロリン脂質、スフィンゴリン脂質、グリセロホスホノ脂質、スフィンゴホスホノ脂質に分類されることもある。 Phospholipids are main components constituting biological membranes widely distributed in animals and plants and microorganisms, and are complex lipids having a structure in which fatty acids, phosphoric acid, bases and the like are bonded to alcohols. Phospholipids are roughly classified into glycerophospholipids containing glycerol and sphingophospholipids containing long-chain amino alcohols such as sphingosine, depending on the type of alcohol bound thereto. Natural phosphorus phospholipids are divided into four groups: glycerophospholipids, sphingophospholipids, glycerophosphonolipids, and sphingophosphonolipids, when the phosphorus atom attachment mode is distinguished from those containing phosphate esters and those containing phosphonic acids. It may be classified into.
グリセロリン脂質の例として、結合している塩基が異なるホスファチジルコリン、ホスファチジルエタノールアミン、ホスファチジルセリン、ホスファチジルイノシトールが挙げられ、塩基がないものとしてホスファチジン酸が挙げられる。スフィンゴリン脂質の例として、スフィンゴミエリンが挙げられる。天然のリン脂質は、結合している脂肪酸の炭素数や二重結合の数が異なるものが種々存在する。また、結合した脂肪酸が１個はずれたリゾ体（リゾリン脂質）も知られている。 Examples of glycerophospholipids include phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylinositol with different bases, and phosphatidic acid having no base. An example of a sphingophospholipid is sphingomyelin. There are various natural phospholipids in which the number of carbon atoms and the number of double bonds of the fatty acid are different. In addition, a lyso form (lysophospholipid) in which one bonded fatty acid is removed is also known.
このように比較的よく知られているリン脂質であっても、その物質構成や構造で区別すると非常に多岐にわたる。近年、その機能が深く研究され、リン脂質は界面活性だけでなく、抗酸化作用、抗癌作用、細胞内外の重要な情報伝達や生命活動に関与するなどの生理活性を有することが判明している。また、リン脂質頭部の極性基と脂肪酸の種類・結合様式及び構成アルコール類による実に多様な分子種がそれぞれ特有の機能を有することが明らかとなってきている。 Even such relatively well-known phospholipids are very diverse when distinguished by their substance structure and structure. In recent years, its functions have been deeply studied, and it has been found that phospholipids have not only surface activity but also physiological activity such as antioxidant activity, anticancer activity, and important information transmission inside and outside cells and life activity. Yes. It has also been clarified that a wide variety of molecular species depending on the polar group of the phospholipid head, the type and binding mode of fatty acids, and the constituent alcohols have specific functions.
現在、リン脂質として産業上、最も多く使用されているものとして大豆リン脂質があり、いわゆる大豆レシチンと呼称され、安価であるため天然乳化剤として重宝されている。その他、卵黄レシチンやイクラ由来のＤＨＡ含有リン脂質（特許文献１）などがあるが、その原料自身が食品として使用されるものであるから、得られたリン脂質は相当高価なものとなり汎用的に使用できないものである。また、これらはいずれもグリセロリン脂質であり、スフィンゴリン脂質はわずかかほとんど含まれていない。 Currently, soybean phospholipid is the most commonly used phospholipid in the industry, so-called soybean lecithin, and it is useful as a natural emulsifier because it is inexpensive. In addition, there are egg yolk lecithin and DHA-containing phospholipids derived from salmon roe (Patent Document 1). However, since the raw materials themselves are used as food, the obtained phospholipids are considerably expensive and versatile. It cannot be used. These are all glycerophospholipids and contain little or almost no sphingophospholipids.
一方、スフィンゴミエリンは動物の脳に多く含まれていることは知られているが、大量生産して汎用的に使用できるまでには至っていない。例えばウシ脳抽出エキスが化粧品に使用されたり、試薬用としてウシの脳や心臓由来のものが販売されている程度である。また、含有量の少ないニワトリ卵黄から高純度に濃縮したものがあるが、やはりコストの高いものとなっている。最近では牛乳由来のスフィンゴミエリン含有リン脂質（特許文献２）が開発され市場に出されているが、やはり高価なため育児用調整粉乳など用途が限定されている。 On the other hand, sphingomyelin is known to be contained in large amounts in animal brains, but has not yet been mass-produced and can be used for general purposes. For example, bovine brain extract is used in cosmetics, or is derived from bovine brain or heart for use as a reagent. Moreover, although there exists what concentrated in high purity from the chicken egg yolk with little content, it is still a high cost thing. Recently, sphingomyelin-containing phospholipids derived from milk (Patent Document 2) have been developed and put on the market. However, since they are expensive, their use such as infant formula milk is limited.
また、構成する脂肪酸に着目したもので、ＤＨＡ含有リン脂質を製造する方法としては、魚介類の加工残渣である頭部や皮や内臓から溶剤抽出する方法（特許文献３〜６）がある。しかしこれらの文献は、ＤＨＡを高含有させることを目的としており、スフィンゴリン脂質を含有させることを示唆するものではない。 In addition, as a method for producing DHA-containing phospholipids, focusing on fatty acids constituting them, there is a method (Patent Documents 3 to 6) for extracting a solvent from the head, skin, or internal organs that are processing residues of seafood. However, these documents are intended to contain DHA in high amounts, and do not suggest that sphingophospholipids are contained.
スフィンゴリン脂質を製造する方法として、醤油油や醤油粕あるいはニワトリの表皮から溶剤抽出する方法（特許文献７〜８）があるが、いずれも非常に煩雑で、品質が良く組成が安定するものを得るには原料の確保と品質維持に多大の努力が必要となり、結局コストがかかるため用途が限定される結果となっている。また、原料に対して得られるリン脂質量は思うほど効率的な収量ではない。結局は安価な原料から、リン脂質を高濃度に含有する、広範囲で汎用性の高いリン脂質組成物を得るには実用性に乏しいものである。 As methods for producing sphingophospholipids, there are methods of solvent extraction from soy sauce oil, soy sauce cake or chicken skin (Patent Documents 7 to 8), all of which are very complicated, have good quality and have a stable composition. In order to obtain it, a great deal of effort is required to secure the raw materials and maintain the quality, and after all costs are high, the use is limited. Moreover, the amount of phospholipid obtained with respect to the raw material is not as efficient as expected. Eventually, it is not practical for obtaining a wide range and highly versatile phospholipid composition containing a high concentration of phospholipid from an inexpensive raw material.
このように飼料から食品、化粧品、試薬、医薬品までの広範囲にわたり、それぞれに利用できるコストに見合って使用できる実用的なリン脂質は今もなお大豆リン脂質だけであり、生理機能の関心の高まりに応じることのできるリン脂質の供給は不十分のままである。また、リン脂質に結合している脂肪酸の違いにより機能が異なることが予想されるため、起源の異なるリン脂質には大いに関心が寄せられている。特にスフィンゴミエリン等のスフィンゴ脂質やＤＨＡ含有リン脂質についてはより一層の研究や活用が期待されている。 In this way, soy phospholipids are still the only practical phospholipid that can be used for a wide range of feeds, foods, cosmetics, reagents, and pharmaceuticals, according to the cost available for each. The supply of available phospholipids remains inadequate. In addition, since the function is expected to be different depending on the fatty acid bound to the phospholipid, phospholipids of different origins are of great interest. In particular, further research and utilization are expected for sphingolipids such as sphingomyelin and DHA-containing phospholipids.
本発明は、商業上利用しやすい原料からスフィンゴミエリンとＤＨＡが結合したグリセロリン脂質を共に含有するリン脂質を提供することを目的とする。さらに、スフィンゴミエリンに結合している脂肪酸に特徴があり、その構造が明確でそれぞれの含有量が調整されたリン脂質を得ることができる方法を提供することも目的とする。 An object of the present invention is to provide a phospholipid containing both glycerophospholipids in which sphingomyelin and DHA are bound together from commercially available raw materials. Furthermore, another object of the present invention is to provide a method capable of obtaining phospholipids characterized by fatty acids bound to sphingomyelin, whose structures are clear and their contents are adjusted.
本発明者は上記課題を解決するために種々検討を重ねた結果、商業的に広く入手が可能であるカツオやマグロなどの魚頭部を主原料として採油された魚原油にリン脂質が多く含有されていること、魚油を精製する前に適切に処理することで効率的にリン脂質を分別・回収できることを見出した。 As a result of various studies to solve the above problems, the present inventor has a large amount of phospholipids in fish crude oil extracted mainly from bonito and tuna fish heads that are widely available commercially. It was found that phospholipids can be efficiently separated and recovered by appropriately treating fish oil before purification.
さらに、得られたリン脂質をアルカリ、有機溶剤などで処理することによりスフィンゴミエリンやグリセロリン脂質の含有量が調整でき、スフィンゴミエリンに結合している脂肪酸の５０％以上が炭素数２０以上という特徴的な脂肪酸であることも見出し、本発明を完成するに至った。 Furthermore, the content of sphingomyelin and glycerophospholipid can be adjusted by treating the obtained phospholipid with an alkali, an organic solvent, etc., and 50% or more of the fatty acids bound to sphingomyelin are characteristically 20 or more carbon atoms. The present invention was also found out to be a perfect fatty acid and completed the present invention.
すなわち、本発明は、
（１）魚頭部を主原料として採油された魚原油から調製されてなるリン脂質組成物であって、スフィンゴミエリンを５％以上、グリセロリン脂質を５％以上含有し、グリセロリン脂質中のＤＨＡ含量が５〜２５％である魚由来リン脂質組成物、
（２）グリセロリン脂質中のホスファチジルコリン及びリゾホスファチジルコリンの合計が９０％以上である（１）のリン脂質組成物、
（３）スフィンゴミエリンを２０〜５０％含有し、グリセロリン脂質を５０〜８０％含有する（１）または（２）のリン脂質組成物、
（４）（１）のリン脂質組成物から調製された、スフィンゴミエリンを９０％以上含有するリン脂質組成物、
（５）スフィンゴミエリンの構成脂肪酸の５０％以上が炭素数２０以上の脂肪酸である（１）〜（４）いずれか１のリン脂質組成物、
（６）スフィンゴミエリンの構成脂肪酸の５０％以上が炭素数２４の脂肪酸である（１）〜（４）いずれか１のリン脂質組成物、
（７）スフィンゴミエリンの構成脂肪酸である炭素数２４の脂肪酸が飽和脂肪酸とモノ不飽和脂肪酸を含み、構成脂肪酸中の炭素数２４モノ不飽和脂肪酸が２５％以上である（１）〜（６）いずれか１のリン脂質組成物、
（８）カツオ及び／またはマグロの頭部を主原料とした魚原油に水和工程を施して沈降した成分を回収することを特徴とする（１）〜（７）いずれか１のリン脂質組成物の製造方法、
（９）水和工程を施して沈降した成分にジアシルグリセロールを０．１〜５％添加してリン脂質組成物と油と水を分離させることを特徴とする（８）のリン脂質組成物の製造方法、
（１０）リン脂質組成物がさらにアルカリ、有機溶剤から選択される１以上の処理に供されることを特徴とする（８）または（９）に記載のリン脂質組成物の製造方法、
（１１）（１）〜（１０）いずれか１のリン脂質組成物を含有する飼料、
を提供するものである。
That is, the present invention
(1) A phospholipid composition prepared from fish crude oil extracted from the fish head as the main raw material, containing 5% or more sphingomyelin and 5% or more glycerophospholipid, and the DHA content in glycerophospholipid A fish-derived phospholipid composition, wherein
(2) The phospholipid composition according to (1), wherein the total of phosphatidylcholine and lysophosphatidylcholine in glycerophospholipid is 90% or more,
(3) The phospholipid composition according to (1) or (2) containing 20 to 50% sphingomyelin and 50 to 80% glycerophospholipid,
(4) A phospholipid composition containing 90% or more of sphingomyelin prepared from the phospholipid composition of (1),
(5) The phospholipid composition according to any one of (1) to (4), wherein 50% or more of the constituent fatty acids of sphingomyelin is a fatty acid having 20 or more carbon atoms,
(6) The phospholipid composition according to any one of (1) to (4), wherein 50% or more of the constituent fatty acids of sphingomyelin is a fatty acid having 24 carbon atoms,
(7) The fatty acid having 24 carbon atoms that is a constituent fatty acid of sphingomyelin contains a saturated fatty acid and a monounsaturated fatty acid, and the monounsaturated fatty acid having 24 carbon atoms in the constituent fatty acid is 25% or more (1) to (6) Any one of the phospholipid compositions,
(8) The phospholipid composition according to any one of (1) to (7), wherein a component that has been precipitated by applying a hydration process to fish crude oil mainly composed of bonito and / or tuna is collected. Manufacturing method,
(9) The phospholipid composition according to (8), wherein 0.1 to 5% of diacylglycerol is added to a component that has been subjected to a hydration step to separate the phospholipid composition, oil and water. Production method,
(10) The method for producing a phospholipid composition according to (8) or (9), wherein the phospholipid composition is further subjected to one or more treatments selected from an alkali and an organic solvent,
(11) A feed containing any one of the phospholipid compositions of (1) to (10),
Is to provide.
本発明によれば、複雑な工程を必要とせずにスフィンゴミエリン、及びＤＨＡが結合したグリセロリン脂質を共に含有するリン脂質を得ることができ、それぞれの含量や結合脂肪酸の特徴による機能を解明する研究や期待される効果を活かした用途として飼料、食品、化粧品、医薬品などに使用できる。 According to the present invention, it is possible to obtain phospholipids containing both sphingomyelin and glycerophospholipid bound to DHA without the need for complicated processes, and to elucidate the functions depending on the contents and characteristics of bound fatty acids. It can be used for feeds, foods, cosmetics, pharmaceuticals, etc. as applications that take advantage of the expected effects.
しかも、本発明で使用される魚原油は商業的に広く入手が可能であるため魚体からの脂肪分抽出の煩雑さが解消される。さらに魚油の精製時に発生するフーツと呼ばれる残滓物からは抽出が困難でしかも品質が低下するが、このような問題も生じない。 Moreover, since the fish crude oil used in the present invention is widely available commercially, the complexity of extracting fat from fish bodies is eliminated. Furthermore, although it is difficult to extract from the residue called foots generated at the time of refining fish oil and the quality is lowered, such a problem does not occur.
本発明は、魚頭部を主原料として採油された魚原油から調製されてなるリン脂質組成物であって、スフィンゴミエリンを５％以上、グリセロリン脂質を５％以上含有し、グリセロリン脂質中のＤＨＡ含量が５〜２５％である魚由来リン脂質組成物である。この組成物のグリセロリン脂質は実質的にホスファチジルコリン及びリゾホスファチジルコリンである。 The present invention is a phospholipid composition prepared from fish crude oil extracted from fish head as a main raw material, containing 5% or more sphingomyelin and 5% or more glycerophospholipid, and DHA in glycerophospholipid It is a fish-derived phospholipid composition having a content of 5 to 25%. The glycerophospholipid of this composition is substantially phosphatidylcholine and lysophosphatidylcholine.
「実質的に」とは、それ以外の成分が含まれないか、含まれていてもわずかか痕跡程度である状態をいう。具体的には、対象となる成分が９０％以上、より好ましくは９５％以上、さらに好ましくは９８％以上である状態をいう。より具体的に説明すると、ホスファチジルコリン及びリゾホスファチジルコリン以外の、ホスファチジルエタノールアミン等が含まれないか、含まれていてもわずかか痕跡程度であり、グリセロリン脂質中のホスファチジルコリン及びリゾホスファチジルコリンの合計が９０％以上、より好ましくは９５％以上、さらに好ましくは９８％以上である状態をいう。 “Substantially” refers to a state in which other components are not included or are slightly or traced even if included. Specifically, it refers to a state in which the target component is 90% or more, more preferably 95% or more, and still more preferably 98% or more. More specifically, phosphatidylcholine and lysophosphatidylcholine other than phosphatidylcholine and lysophosphatidylcholine are not included, or even if they are included, the amount of phosphatidylcholine and lysophosphatidylcholine in glycerophospholipid is 90% or more. More preferably 95% or more, and still more preferably 98% or more.
本発明のリン脂質を得るための原料となる魚原油は魚頭部を主体として採油されたものである。具体的にはカツオやマグロの頭部から採油された魚油が望ましい。なお、採油する頭部に他の部位、例えば尾部等が少量混入していてもかまわない。例えばカツオの場合、鹿児島県の枕崎や静岡県の焼津等で鰹節等に使用した可食部以外の頭部を主体とする残渣から大規模な採油がおこなわれているが、このような魚原油を利用できる。例えばマグロの場合、生食や缶詰に使用した可食部以外の頭部を主体とする残渣から大規模な採油がおこなわれているが、このような魚原油を利用できる。魚原油はリン脂質を原油に対して０．１％以上含有するものが望ましい。イワシ油やサンマ油、サバ油のように全魚体から採油した原油はリン脂質の含量が少ないため好ましくはない。魚原油の採油は通常の工業的に行なわれる方法、すなわち連続式加熱圧搾法（クッカー・プレス）による搾汁を遠心分離し、油層部を回収する方法等で行われる。 The fish crude oil, which is a raw material for obtaining the phospholipid of the present invention, is extracted mainly from the fish head. Specifically, fish oil extracted from the head of skipjack or tuna is desirable. In addition, other parts, for example, a tail part etc., may mix in the head which collects oil. For example, in the case of skipjack, large-scale oil is collected from residues mainly in the head other than the edible portion used for bonito in Makurazaki in Kagoshima Prefecture and Yaizu in Shizuoka Prefecture. Can be used. For example, in the case of tuna, large-scale oil is collected from a residue mainly composed of the head other than the edible portion used for raw food and canned food, and such fish crude oil can be used. The fish crude oil preferably contains 0.1% or more of phospholipid relative to the crude oil. Crude oil collected from whole fish, such as sardine oil, saury oil and mackerel oil, is not preferred because of its low phospholipid content. Fish crude oil is collected by an ordinary industrial method, that is, a method of centrifuging squeezed juice by a continuous heat pressing method (Cooker press) and recovering the oil layer portion.
本発明のリン脂質組成物のスフィンゴミエリン含量は５％以上である。この含量は、リン脂質組成物の用途等に応じて適宜調整することができ、例えば、１０％以上、１５％以上、２０％以上、３０％以上、４０％以上、５０％以上、６０％以上、７０％以上、８０％以上、９０％以上とすることができる。本発明のリン脂質組成物のスフィンゴミエリン含量の好ましい例は、５〜２０、５〜３０、５〜４０、５〜５０、５〜６０、５〜７０、５〜８０、５〜９０、５〜９５、１０〜２０、１０〜３０、１０〜４０、１０〜５０、１０〜６０、１０〜７０、１０〜８０、１０〜９０、２０〜３０、２０〜４０、２０〜５０、２０〜６０、２０〜７０、２０〜８０、２０〜９０、３０〜４０、３０〜５０、３０〜６０、３０〜７０、３０〜８０、３０〜９０、４０〜５０、４０〜６０、４０〜７０、４０〜８０、４０〜９０、５０〜６０、５０〜７０、５０〜８０、５０〜９０、６０〜７０、６０〜８０、６０〜９０、７０〜８０、７０〜９０、８０〜９０％などである。 The sphingomyelin content of the phospholipid composition of the present invention is 5% or more. This content can be appropriately adjusted according to the use of the phospholipid composition, for example, 10% or more, 15% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more. 70% or more, 80% or more, or 90% or more. Preferred examples of the sphingomyelin content of the phospholipid composition of the present invention are 5-20, 5-30, 5-40, 5-50, 5-60, 5-70, 5-80, 5-90, 5 95, 10-20, 10-30, 10-40, 10-50, 10-60, 10-70, 10-80, 10-90, 20-30, 20-40, 20-50, 20-60, 20-70, 20-80, 20-90, 30-40, 30-50, 30-60, 30-70, 30-80, 30-90, 40-50, 40-60, 40-70, 40- 80, 40 to 90, 50 to 60, 50 to 70, 50 to 80, 50 to 90, 60 to 70, 60 to 80, 60 to 90, 70 to 80, 70 to 90, 80 to 90%, and the like.
本発明のリン脂質組成物のグリセロリン脂質含量は５％以上である。この含量は、リン脂質組成物の用途等に応じて適宜調整することができ、例えば、１０％以上、１５％以上、２０％以上、３０％以上、４０％以上、５０％以上、６０％以上、７０％以上、８０％以上、９０％以上とすることができる。本発明のリン脂質組成物のグリセロリン脂質含量の好ましい例は、５〜２０、５〜３０、５〜４０、５〜５０、５〜６０、５〜７０、５〜８０、５〜９０、５〜９５、１０〜２０、１０〜３０、１０〜４０、１０〜５０、１０〜６０、１０〜７０、１０〜８０、１０〜９０、２０〜３０、２０〜４０、２０〜５０、２０〜６０、２０〜７０、２０〜８０、２０〜９０、３０〜４０、３０〜５０、３０〜６０、３０〜７０、３０〜８０、３０〜９０、４０〜５０、４０〜６０、４０〜７０、４０〜８０、４０〜９０、５０〜６０、５０〜７０、５０〜８０、５０〜９０、６０〜７０、６０〜８０、６０〜９０、７０〜８０、７０〜９０、８０〜９０％などである。 The glycerophospholipid content of the phospholipid composition of the present invention is 5% or more. This content can be appropriately adjusted according to the use of the phospholipid composition, for example, 10% or more, 15% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more. 70% or more, 80% or more, or 90% or more. Preferred examples of the glycerophospholipid content of the phospholipid composition of the present invention are 5 to 20, 5 to 30, 5 to 40, 5 to 50, 5 to 60, 5 to 70, 5 to 80, 5 to 90, and 5 to 5. 95, 10-20, 10-30, 10-40, 10-50, 10-60, 10-70, 10-80, 10-90, 20-30, 20-40, 20-50, 20-60, 20-70, 20-80, 20-90, 30-40, 30-50, 30-60, 30-70, 30-80, 30-90, 40-50, 40-60, 40-70, 40- 80, 40 to 90, 50 to 60, 50 to 70, 50 to 80, 50 to 90, 60 to 70, 60 to 80, 60 to 90, 70 to 80, 70 to 90, 80 to 90%, and the like.
本発明のリン脂質組成物において、グリセロリン脂質中のＤＨＡ含量は５〜２５％である。この含量は、リン脂質組成物の用途等に応じて適宜調整することができる。グリセロリン脂質中のＤＨＡ含量の好ましい例は、５〜１０、５〜１５、５〜２０、５〜２５、１０〜１５、１０〜２０、１０〜２５、１５〜２０、１５〜２５、２０〜２５％などである。 In the phospholipid composition of the present invention, the DHA content in the glycerophospholipid is 5 to 25%. This content can be appropriately adjusted according to the use of the phospholipid composition. Preferred examples of the DHA content in the glycerophospholipid are 5 to 10, 5 to 15, 5 to 20, 5 to 25, 10 to 15, 10 to 20, 10 to 25, 15 to 20, 15 to 25, and 20 to 25. %.
本発明のリン脂質組成物において、グリセロリン脂質は実質的にホスファチジルコリン及びリゾホスファチジルコリンである。また、グリセロリン脂質中のホスファチジルコリン：リゾホスファチジルコリン含量はリン脂質組成物の用途等に応じて任意の比率で適宜調整することができる。例えば、ホスファチジルコリン４０〜９０％：リゾホスファチジルコリン１０〜５０％としたり、ホスファチジルコリン３０％以下：リゾホスファチジルコリン７０％以上としたりすることもできる。 In the phospholipid composition of the present invention, the glycerophospholipid is substantially phosphatidylcholine and lysophosphatidylcholine. The phosphatidylcholine: lysophosphatidylcholine content in the glycerophospholipid can be appropriately adjusted at an arbitrary ratio depending on the use of the phospholipid composition. For example, phosphatidylcholine may be 40 to 90%: lysophosphatidylcholine 10 to 50%, or phosphatidylcholine 30% or less: lysophosphatidylcholine 70% or more.
本発明のリン脂質組成物において、スフィンゴミエリンの構成脂肪酸の５０％以上は炭素数２０以上の脂肪酸であり、好ましくは構成脂肪酸の５０％以上が炭素数２２以上の脂肪酸であり、さらに好ましくは構成脂肪酸の５０％以上が炭素数２４の脂肪酸である。さらに好ましくは、炭素数２４の構成脂肪酸が飽和脂肪酸とモノ不飽和脂肪酸であり、構成脂肪酸中の炭素数２４モノ不飽和脂肪酸が２５％以上である。 In the phospholipid composition of the present invention, 50% or more of the constituent fatty acids of sphingomyelin is a fatty acid having 20 or more carbon atoms, preferably 50% or more of the constituent fatty acids are fatty acids having 22 or more carbon atoms, more preferably a constituent. More than 50% of the fatty acids are fatty acids having 24 carbon atoms. More preferably, the constituent fatty acid having 24 carbon atoms is a saturated fatty acid and a monounsaturated fatty acid, and the monounsaturated fatty acid having 24 carbon atoms in the constituent fatty acid is 25% or more.
続いて、本発明のリン脂質組成物の製造方法について説明する。本発明のリン脂質組成物の製造方法は、魚原油を精製する前に処理を行うことを特徴とする。アルカリ脱酸等の精製後の油からではリン脂質が除去されてしまうのでリン脂質を得ることはできない。また、精製時に発生するフーツと呼ばれる油滓から回収しようとすると効率が悪く、品質の悪いものとなる。 Then, the manufacturing method of the phospholipid composition of this invention is demonstrated. The method for producing a phospholipid composition of the present invention is characterized in that the treatment is performed before refining fish crude oil. Since the phospholipid is removed from the oil after purification such as alkali deoxidation, the phospholipid cannot be obtained. Moreover, if it tries to collect | recover from the oil bottle called foots which generate | occur | produces at the time of refinement | purification, it will become inefficient and bad quality.
まず、魚原油の夾雑物を遠心分離またはろ過等の処理により除くことが望ましい。魚原油には重合物や微量の蛋白等の固形物が混在することがあり、これらがあると得られたリン脂質の品質が低下する。次に魚原油を油脂結晶が融解しており、かつできるだけ低い温度とした上で水を添加する。この温度として３０〜４０℃が望ましい。水の添加量は予想されるリン脂質の含有量により調整でき、魚原油に対して１〜５％、望ましくは２〜４％が好ましい。添加水の温度は油温と同じか、近い温度が好ましい。水添加後、激しく攪拌することでリン脂質の油からの遊離と凝集を促進する。さらに、添加する水粒子をできるだけ小さくして油中に供給すると、さらに遊離と凝集が促進され好ましい。したがって微細な孔径の膜を通して加水するなどの公知の方法も有効である。撹拌の後、静置すると上部に油層、中間部にリン脂質層、下部に水層となる３層に分離するので中間層を回収する。この時に遠心分離機を使用することができる。飼料用であれば後に水分や油を加える場合もあるのでこのようにして得られたリン脂質をそのまま利用できる。汎用的に使用するのであれば、工業的に公知の方法で水分を除去することが好ましい。また、残存する中性油を除くことでより良い品質のリン脂質組成物が得られるため、中性油、すなわちアシルグリセロールを更なる遠心分離、アセトン分配等により除去することが好ましい。この場合、アセトンを使用すれば水分も同時に除去が可能となる。さらにヘキサンやエタノールなどの有機溶剤を併用することもできる。 First, it is desirable to remove contaminants of fish crude oil by a process such as centrifugation or filtration. Fish crude oil may contain a solid matter such as a polymer or a trace amount of protein, and if these are present, the quality of the obtained phospholipid deteriorates. Next, water is added after the fat and oil crystals are melted in the fish crude oil and the temperature is made as low as possible. As this temperature, 30-40 degreeC is desirable. The amount of water added can be adjusted by the expected content of phospholipid, and is preferably 1 to 5%, desirably 2 to 4%, relative to fish crude oil. The temperature of the added water is preferably the same as or close to the oil temperature. After the addition of water, vigorous stirring promotes the release and aggregation of phospholipids from the oil. Further, it is preferable that the water particles to be added are made as small as possible and are supplied into the oil because liberation and aggregation are further promoted. Therefore, a known method such as adding water through a membrane having a fine pore diameter is also effective. After stirring, the mixture is allowed to stand to separate into three layers, an oil layer at the top, a phospholipid layer at the middle, and an aqueous layer at the bottom, so the middle layer is recovered. At this time, a centrifuge can be used. If it is for feed, water or oil may be added later, so the phospholipid thus obtained can be used as it is. If it is used for general purposes, it is preferable to remove moisture by a method known in the industry. Further, since a phospholipid composition of better quality can be obtained by removing the remaining neutral oil, it is preferable to remove the neutral oil, that is, acylglycerol, by further centrifugation, acetone distribution, or the like. In this case, if acetone is used, moisture can be removed at the same time. Further, an organic solvent such as hexane or ethanol can be used in combination.
さらに分取した前記リン脂質層に対してジアシルグリセロールを０．１〜５％添加して攪拌後静置し、前記同様中間層を回収すると混在する水や油を効率的に除去しリン脂質含量の高いものを得ることができる。ジアシルグリセロールとしては純度１００％のもの、または１０〜９９％含有される油脂を用いることができる。ジアシルグリセロールを得る方法としては油脂を加水分解する方法、グリセリンと脂肪酸あるいはそのエステルから合成する方法等公知の方法を用いることができる。得られたリン脂質は前記と同様に水分除去や中性油除去を行うことが好ましい。 Furthermore, 0.1 to 5% of diacylglycerol is added to the separated phospholipid layer, and the mixture is allowed to stand after stirring, and when the intermediate layer is recovered, the mixed water and oil are efficiently removed and the phospholipid content is reduced. Can be obtained. As the diacylglycerol, those having a purity of 100% or oils containing 10 to 99% can be used. As a method for obtaining diacylglycerol, known methods such as a method of hydrolyzing fats and oils, a method of synthesizing from glycerin and a fatty acid or an ester thereof can be used. The obtained phospholipid is preferably subjected to water removal and neutral oil removal as described above.
このようにして得られたリン脂質組成物はスフィンゴミエリン、及びＤＨＡが結合したグリセロリン脂質を含有する本発明の魚由来リン脂質組成物となる。スフィンゴミエリンを５％以上、グリセロリン脂質を５％以上含有するスフィンゴミエリン含有リン脂質組成物である。この組成物のグリセロリン脂質は実質的にホスファチジルコリン及びリゾホスファチジルコリンである。 The phospholipid composition thus obtained is the fish-derived phospholipid composition of the present invention containing sphingomyelin and glycerophospholipid bound to DHA. A sphingomyelin-containing phospholipid composition containing 5% or more sphingomyelin and 5% or more glycerophospholipid. The glycerophospholipid of this composition is substantially phosphatidylcholine and lysophosphatidylcholine.
得られたリン脂質組成物にはまだ脂肪酸や中性油さらにスフィンゴ糖脂質やその他未同定の脂溶性物質が含まれている。従って、本発明の別の態様では、使用する用途に応じて、さらに純度を上げたり、特定の構成物質の含量を高めることができる。例えば、有機溶剤を使用しカラム分画することで、グリセロリン脂質を４０％以上でスフィンゴミエリンを２０％以上にすることもできる。更には、例えば、カセイソーダ等のアルカリ水溶液で加水分解を行い、グリセロリン脂質を分解することでスフィンゴミエリン含有量の高いリン脂質組成物とすることができる。この場合、例えばスフィンゴミエリンを９０％以上含有し、グリセロリン脂質が１０％以下である本発明のスフィンゴミエリン高含有リン脂質組成物が得られる。より詳細には、リン脂質が実質的にスフィンゴミエリンであるスフィンゴミエリン高含有リン脂質組成物が得られる。 The resulting phospholipid composition still contains fatty acids, neutral oils, glycosphingolipids and other unidentified fat-soluble substances. Therefore, in another aspect of the present invention, the purity can be further increased or the content of a specific constituent material can be increased depending on the application to be used. For example, by performing column fractionation using an organic solvent, it is possible to make glycerophospholipid 40% or more and sphingomyelin 20% or more. Furthermore, it can be set as a phospholipid composition with high sphingomyelin content by hydrolyzing with alkaline aqueous solutions, such as caustic soda, and decomposing | disassembling glycerophospholipid. In this case, for example, the sphingomyelin-rich phospholipid composition of the present invention containing 90% or more of sphingomyelin and 10% or less of glycerophospholipid is obtained. More specifically, a sphingomyelin-rich phospholipid composition in which the phospholipid is substantially sphingomyelin is obtained.
次に実施例を示して本発明をさらに詳しく、具体的に説明する。なお、リン脂質組成物の各構成成分の測定は以下の方法で行なった。 Next, the present invention will be described in more detail and specifically with reference to examples. In addition, the measurement of each structural component of a phospholipid composition was performed with the following method.
総リン脂質含量の測定方法
Ｂａｒｔｌｅｔｔの方法（Ｂａｒｔｌｅｔｔ，Ｇ．Ｒ．，Ｊ．Ｂｉｏｌ．Ｃｈｅｍ．，２３４，４６６（１９５９））でリン量を定量し、リン脂質量に換算した。
Method for Measuring Total Phospholipid Content The amount of phospholipid was quantified by the method of Bartlett (Bartlett, GR, J. Biol. Chem., 234, 466 (1959)) and converted to the amount of phospholipid.
リン脂質組成物をクロロホルム／メタノール混合液（１／１）に溶解し、試料を１０ｍｇ／ｍｌの濃度に調整した。調製した試料を１００μｌ試験管に取り溶媒を留去した後、１０Ｎの硫酸を１ｍｌ加え、２３０℃で３時間加熱処理を行った。次いで３０％過酸化水素水を２滴加え、１５０℃で１．５時間加熱処理を行った。このものを放冷した後０．２２％モリブデン酸アンモニウム溶液９．２ｍｌとＦｉｓｋｅ Ｓｕｂｂａｒｏｗ試薬０．４ｍｌを加え、沸騰水中で７分間加熱した。次にろ紙濾過を行い蒸留水で２５ｍｌにメスアップしたのち、U−２０００ダブルビーム分光光度計（日立製作所製）で波長８３０ｎｍの吸光度を測定した。得られた吸光度からリン含量を算出し、さらにステアロイル−オレイル−ホスファチジルコリン（分子量７９０）に相当するものとして、リン脂質含量に換算して総リン脂質含量とした。なお、リン濃度と吸光度の関係、吸光度からのリン脂質含量の算出式は下記に示した。 The phospholipid composition was dissolved in a chloroform / methanol mixture (1/1), and the sample was adjusted to a concentration of 10 mg / ml. The prepared sample was placed in a 100 μl test tube, the solvent was distilled off, 1 ml of 10N sulfuric acid was added, and heat treatment was performed at 230 ° C. for 3 hours. Next, 2 drops of 30% hydrogen peroxide water was added, and heat treatment was performed at 150 ° C. for 1.5 hours. After standing to cool, 9.2 ml of 0.22% ammonium molybdate solution and 0.4 ml of Fiske Subbarow reagent were added and heated in boiling water for 7 minutes. Next, after filtering with filter paper and making up to 25 ml with distilled water, absorbance at a wavelength of 830 nm was measured with a U-2000 double beam spectrophotometer (manufactured by Hitachi, Ltd.). The phosphorus content was calculated from the obtained absorbance, and the total phospholipid content was calculated in terms of phospholipid content as equivalent to stearoyl-oleyl-phosphatidylcholine (molecular weight 790). The relationship between the phosphorus concentration and the absorbance, and the calculation formula for the phospholipid content from the absorbance are shown below.
リン濃度と吸光度の関係
１０μｇ／ｍｌ濃度のＫＨ２ＰＯ４標準液を調製し、０、０．５、１．０、２．０、３．０ｍｌずつ試験管に取り上記と同様の分析を行い、リン濃度と吸光度の検量線を作成した。リン濃度と吸光度の関係は下記の通りであった。
検量線 ｙ＝２６．９ｘ＋０．０１５８
ｙ：［リン濃度（μｇ／ｍｌ）］
ｘ：［吸光度］
Relationship between phosphorus concentration and absorbance Prepare a 10 μg / ml KH 2 PO 4 standard solution, and place 0, 0.5, 1.0, 2.0, and 3.0 ml each in a test tube and perform the same analysis as above. A calibration curve of phosphorus concentration and absorbance was prepared. The relationship between phosphorus concentration and absorbance was as follows.
Calibration curve y = 26.9x + 0.0158
y: [phosphorus concentration (μg / ml)]
x: [Absorbance]
総リン脂質含量の算出
リン脂質としてリン含量からリン脂質への質量換算は定法に従い係数２５を用いた。また、純度既知の大豆リン脂質で同様の分析操作を行い、回収率の補正を加えた。
［リン脂質含量］=（２６．９×［吸光度］＋０．０１５８）×２５／［回収率］／１０００×１００
Calculation of the total phospholipid content As a phospholipid, a mass conversion from a phosphorus content to a phospholipid was performed using a coefficient of 25 according to a conventional method. In addition, the same analysis operation was performed with soybean phospholipid of known purity, and the recovery rate was corrected.
[Phospholipid content] = (26.9 × [absorbance] +0.0158) × 25 / [recovery] / 1000 × 100
グリセロリン脂質の各組成含量及びスフィンゴミエリン含量の測定方法
薄層クロマトグラフィーで各リン脂質成分を分離させた後、ＦＩＤ検出器による定量をイアトロスキャンＭｋ−６（三菱化学ヤトロン社）を用いて行なった。分離された各リン脂質成分のホスファチジルコリン、ホスファチジルエタノールアミンの総計をリゾ体も含めてグリセロリン脂質とした。
Measuring method of each composition content and sphingomyelin content of glycerophospholipid After separating each phospholipid component by thin layer chromatography, quantification with an FID detector is performed using Iatroscan Mk-6 (Mitsubishi Chemical Yatron). It was. The total of the separated phosphatidylcholine and phosphatidylethanolamine of each phospholipid component, including the lyso form, was designated as glycerophospholipid.
具体的にはリン脂質組成物をクロロホルム／メタノール混合液（１／１）に溶解し、試料を１０ｍｇ／ｍｌの濃度に調整した。続いて試料を５μｌクロマロッド（登録商標）にスポットし、クロロホルム／メタノール／水／アンモニア水（４７／２０／２．５／０．２５）の混合溶媒で１５分間展開した後、クロマロッドを乾燥させ、その後同様の混合溶媒で５０分間の展開を行った。リン脂質各スポットは、市販品の各リン脂質標準品を同様に分離展開し、そのＲｆ値から判別した。前記の総リン脂質含量にイアトロスキャンで得られた各リン脂質成分の組成百分率を掛けてグリセロリン脂質の各組成含量及びスフィンゴミエリン含量とした。 Specifically, the phospholipid composition was dissolved in a chloroform / methanol mixture (1/1), and the sample was adjusted to a concentration of 10 mg / ml. Subsequently, the sample was spotted on a 5 μl Chromarod (registered trademark), developed with a mixed solvent of chloroform / methanol / water / ammonia water (47/20 / 2.5 / 0.25) for 15 minutes, and then the chromarod was dried. Then, development for 50 minutes was performed with the same mixed solvent. For each phospholipid spot, each phospholipid standard product on the market was separated and developed in the same manner, and was discriminated from its Rf value. The total phospholipid content was multiplied by the composition percentage of each phospholipid component obtained by Iatroscan to obtain each composition content and sphingomyelin content of glycerophospholipid.
なお、リン脂質各スポットのＲｆ値の判別に使用した標準品は以下の通りである。
ホスファチジルコリン（大豆由来・フナコシ社製）、リゾホスファチジルコリン（卵黄由来・和光純薬工業社製）、スフィンゴミエリン（牛脳由来・シグマアルドリッチ社製）、ホスファチジルエタノールアミン（卵由来・和光純薬工業社製）、リゾホスファチジルエタノールアミン（卵由来・フナコシ社製）、ホスファチジン酸（卵由来・フナコシ社製）、ホスファチジルセリン（牛脳由来・フナコシ社製）、ホスファチジルイノシトール（ブタ肝臓由来・フナコシ社製）。
In addition, the standard goods used for discrimination | determination of Rf value of each phospholipid spot are as follows.
Phosphatidylcholine (derived from soybean, manufactured by Funakoshi), lysophosphatidylcholine (derived from egg yolk, manufactured by Wako Pure Chemical Industries), sphingomyelin (derived from bovine brain, manufactured by Sigma Aldrich), phosphatidylethanolamine (derived from egg, manufactured by Wako Pure Chemical Industries, Ltd.) ), Lysophosphatidylethanolamine (egg derived from Funakoshi), phosphatidic acid (egg derived from Funakoshi), phosphatidylserine (cattle brain derived from Funakoshi), phosphatidylinositol (pig liver derived from Funakoshi).
グリセロリン脂質の構成脂肪酸とその含量の測定方法
リン脂質組成物のアセトン可溶成分、即ち中性脂質や脂肪酸をアセトン洗浄により除去した。さらに得られたアセトン処理リン脂質組成物中から、分取用薄層クロマトグラフィープレート（メルク社製）を用いてグリセロリン脂質のみを集めた。すなわち、アセトン処理リン脂質組成物をクロロホルム／メタノール／水（４７／２０／２．５）の混合溶媒により展開分離し、グリセロリン脂質相当画分のみを掻き取り収集した。グリセロリン脂質のメチル化は、三フッ化ホウ素−メタール法により行った。得られた脂肪酸のメチルエステルを常法のガスクロマトグラフィー法により分析した。用いた装置はＧＣ−１７Ａ（島津製作所社製）、カラムはＢＰＸ−７０（ＳＧＥ社製）、昇温条件は、１４０℃で２分保持後、２４０℃まで５℃／分で昇温し、２４０℃で２０分保持とした。
Measuring method of constituent fatty acid and content of glycerophospholipid Acetone-soluble components of the phospholipid composition, that is, neutral lipid and fatty acid were removed by washing with acetone. Furthermore, only glycerophospholipid was collected from the obtained acetone-treated phospholipid composition using a preparative thin layer chromatography plate (manufactured by Merck). That is, the acetone-treated phospholipid composition was developed and separated with a mixed solvent of chloroform / methanol / water (47/20 / 2.5), and only the glycerophospholipid-equivalent fraction was scraped and collected. The methylation of glycerophospholipid was performed by a boron trifluoride-methal method. The obtained fatty acid methyl ester was analyzed by a conventional gas chromatography method. The apparatus used was GC-17A (manufactured by Shimadzu Corporation), the column was BPX-70 (manufactured by SGE), and the temperature raising conditions were held at 140 ° C. for 2 minutes and then raised to 240 ° C. at 5 ° C./min. It was kept at 240 ° C. for 20 minutes.
スフィンゴミエリンの構成脂肪酸とその含量の測定方法
スフィンゴミエリンの構成脂肪酸組成を以下の方法で測定した。まずリン脂質組成物のアセトン可溶成分をアセトン洗浄により除去し、得られたアセトン処理リン脂質組成物中からスフィンゴミエリンを単離した。アセトン処理して得られたリン脂質組成物を０．４Ｎ ＫＯＨ−メタノール溶液で処理しグリセロリン脂質の加水分解を行った後、シリカゲルカラム精製によりスフィンゴミエリン画分を回収した。
次に精製スフィンゴミエリン画分１ｇに１Ｎ ＨＣｌ／メタノール２０ｍｌを加え９０℃で一晩静置し、スフィンゴミエリン構成脂肪酸のメチル化を行った。反応液をエバポレーターで濃縮した後、冷アセトン２０ｍｌを加え撹拌し、冷アセトン可溶部を脂肪酸メチルエステル画分として回収した。脂肪酸組成は常法のガスクロマトグラフィー法により分析した。用いた装置はＧＣ−１７Ａ（島津製作所社製）、カラムはＢＰＸ−７０（ＳＧＥ社製）、昇温条件は、１４０℃で２分保持後、２４０℃まで５℃／分で昇温し、２４０℃で２０分保持とした。
Method for measuring constituent fatty acids of sphingomyelin and its content The constituent fatty acid composition of sphingomyelin was measured by the following method. First, acetone-soluble components of the phospholipid composition were removed by washing with acetone, and sphingomyelin was isolated from the obtained acetone-treated phospholipid composition. The phospholipid composition obtained by the acetone treatment was treated with a 0.4N KOH-methanol solution to hydrolyze glycerophospholipid, and then a sphingomyelin fraction was collected by silica gel column purification.
Next, 20 ml of 1N HCl / methanol was added to 1 g of the purified sphingomyelin fraction, and the mixture was allowed to stand at 90 ° C. overnight to methylate the sphingomyelin constituent fatty acid. After concentrating the reaction solution with an evaporator, 20 ml of cold acetone was added and stirred, and the cold acetone soluble part was recovered as a fatty acid methyl ester fraction. The fatty acid composition was analyzed by a conventional gas chromatography method. The apparatus used was GC-17A (manufactured by Shimadzu Corporation), the column was BPX-70 (manufactured by SGE), and the temperature raising conditions were held at 140 ° C. for 2 minutes and then raised to 240 ° C. at 5 ° C./min. It was kept at 240 ° C. for 20 minutes.
また、上記の操作で得られたアセトン処理リン脂質組成物がスフィンゴミエリンを含むことを確認するため、ＬＣ／ＭＳ Ｆｉｎｎｉｇａｎ ＬＣＱＤＥＣＡ（Ｔｈｅｒｍｏ Ｑｕｅｓｔ社製）により構造確認を行った。上記精製スフィンゴミエリンをクロロホルム／メタノール（１／１）混合液に溶解し、ＬＣ／ＭＳ分析サンプルとした。カラムは５ＳＬ−ＩＩ（４．０ｍｍ×１５０ｍｍ）（ナカライテスク社製）を用い、ポジティブモードで分析を行った（ｍ／ｚの値はＮａ付加体：分子量＋２３）。スフィンゴミエリンの構成脂肪酸としてパルミチン酸（Ｃ１６：０、分子量２３９．４）が結合したｍ／ｚ＝７２５．６のピークが確認できた。同様にベヘン酸（Ｃ２２：０、分子量３４０．６）、リグノセリン酸（Ｃ２４：０、分子量３６８．６）、ネルボン酸（Ｃ２４：１、分子量３６６．６）が結合したスフィンゴミエリンに相当するｍ／ｚ＝８０９．６、ｍ／ｚ＝８３７．６、ｍ／ｚ＝８３５．６のピークをそれぞれ確認した。また上記精製スフィンゴミエリン酸分解物にて、下記（Ｉ）のＲ＝Ｈである化合物に相当するｍ／ｚ＝４８７．２のピークを確認した。これらのことから、シリカゲル精製により得られたリン脂質が、下記に示す構造のスフィンゴミエリンを含むことが確認できた。
実施例１ カツオ原油からのリン脂質組成物の調製
カツオ原油（カツオ頭部を主体とする鰹節製造時の残渣から採油された魚原油、酸価８．５）1０ｋｇを６０℃に加温した後、３５℃まで下げ、３００ｇの水温３５℃の水を滴下しながら１５分間攪拌し３時間静置後４５００ｒｐｍで遠心分離を行い中間層を集めた。この中間層に５℃のアセトン２０００ｍｌを混合した後、アセトン不溶性画分を分取し、アセトンを真空下で留去し、リン脂質組成物４０ｇを得た。得られたリン脂質組成物は、総リン脂質含量は３５％であり、スフィンゴミエリンを１１．０％、グリセロリン脂質を２４．０％含有し、グリセロリン脂質中のＤＨＡ含量が９．５％であった。また、グリセロリン脂質中のホスファチジルコリンは７２．３％、リゾホスファチジルコリンは２６．４％で、合計で９８．７％であるのに対しホスファチジルエタノールアミンはわずか１．３％であり、その他のグリセロリン脂質は検出されなかった。
カツオ原油の脂肪酸組成及び得られたリン脂質組成物中のグリセロリン脂質の構成脂肪酸組成を表１に示す。また、得られたリン脂質組成物中のスフィンゴミエリンの構成脂肪酸組成を表２に示す。
Example 1 Preparation of Phospholipid Composition from Skipjack Crude Oil After heating 10 kg of bonito crude oil (fish crude oil collected from residue during bonito manufacturing, mainly bonito head, acid value 8.5) to 60 ° C The temperature was lowered to 35 ° C., 300 g of water having a water temperature of 35 ° C. was dropped, and the mixture was stirred for 15 minutes, allowed to stand for 3 hours, and centrifuged at 4500 rpm to collect an intermediate layer. After mixing 2000 ml of acetone at 5 ° C. with this intermediate layer, the acetone insoluble fraction was collected, and acetone was distilled off under vacuum to obtain 40 g of a phospholipid composition. The resulting phospholipid composition had a total phospholipid content of 35%, 11.0% sphingomyelin, 24.0% glycerophospholipid, and a DHA content of 9.5% in glycerophospholipid. It was. In addition, phosphatidylcholine in glycerophospholipid is 72.3%, lysophosphatidylcholine is 26.4%, and the total is 98.7%, whereas phosphatidylethanolamine is only 1.3%, and other glycerophospholipids are Not detected.
Table 1 shows the fatty acid composition of bonito crude oil and the constituent fatty acid composition of glycerophospholipid in the obtained phospholipid composition. Table 2 shows the constituent fatty acid composition of sphingomyelin in the obtained phospholipid composition.
実施例２ マグロ原油からのリン脂質組成物の調製
実施例１においてカツオ原油のかわりにマグロ由来原油（主にマグロ残渣から採油された魚原油、酸価６．７）１０ｋｇを用いた以外は同様の操作を行い、リン脂質組成物５４ｇを得た。得られたリン脂質組成物は、総リン脂質含量３２％であり、スフィンゴミエリンを９．５％、グリセロリン脂質を２２．５％含有し、グリセロリン脂質中のＤＨＡ含量が１２．４％であった。また、グリセロリン脂質中のホスファチジルコリンは７９．０％、リゾホスファチジルコリンは２０．１％で、合計で９９．１％であるのに対しホスファチジルエタノールアミンはわずか０．９％であり、その他のグリセロリン脂質は検出されなかった。
得られたリン脂質組成物中のグリセロリン脂質の構成脂肪酸組成を表１に示す。
Example 2 Preparation of Phospholipid Composition from Tuna Crude Oil Same as Example 1 except that 10 kg of tuna-derived crude oil (mainly fish crude oil extracted from tuna residue, acid value 6.7) was used instead of skipjack crude oil. Thus, 54 g of a phospholipid composition was obtained. The obtained phospholipid composition had a total phospholipid content of 32%, a sphingomyelin content of 9.5%, a glycerophospholipid content of 22.5%, and a DHA content in the glycerophospholipid of 12.4%. . In addition, phosphatidylcholine in glycerophospholipid is 79.0%, lysophosphatidylcholine is 20.1%, the total is 99.1%, while phosphatidylethanolamine is only 0.9%, and other glycerophospholipids are Not detected.
Table 1 shows the constituent fatty acid composition of glycerophospholipid in the obtained phospholipid composition.
比較例1
実施例１においてカツオ原油のかわりにメンヘーデン原油（メンヘーデンから採油された魚原油、酸価６．５）１０ｋｇを用いた以外は同様の操作を行ったが、リン脂質組成物はわずか９ｇしか得られなかった。また、得られたリン脂質組成物中の総リン脂質含量は１０％であった。
Comparative Example 1
The same operation was carried out except that 10 kg of Menhaden crude oil (fish crude oil extracted from Menhaden, acid value 6.5) was used in place of bonito crude oil in Example 1, but only 9 g of phospholipid composition was obtained. There wasn't. Moreover, the total phospholipid content in the obtained phospholipid composition was 10%.
比較例２
実施例１においてカツオ原油のかわりにイワシ原油（主にイワシから採油された魚原油、酸価７．４）１０ｋｇを用いた以外は同様の操作を行ったが、リン脂質組成物は得られなかった。
Comparative Example 2
The same operation was performed except that 10 kg of sardine crude oil (mainly fish crude oil extracted from sardines, acid value 7.4) was used in place of bonito crude oil in Example 1, but no phospholipid composition was obtained. It was.
実施例３ リン脂質高含有リン脂質組成物の調製
実施例１で得たリン脂質組成物２ｇをクロロホルムに溶解し、２００ｇのシリカゲル（ワコーゲルＣ−２００、和光純薬工業社製）を充填したガラスカラムでのケイ酸カラムクロマトグラフィーにより精製を行い、リン脂質相当画分のみを分離分取した。クロロホルム６００ｍｌで中性脂質画分を洗い流した後、メタノール１０００ｍｌでリン脂質画分を溶出させ、溶剤を留去しリン脂質高含有リン脂質組成物１ｇを得た。得られたリン脂質組成物は、総リン脂質含量９１％であり、スフィンゴミエリンを３３％、グリセロリン脂質を５８％含有し、グリセロリン脂質中のＤＨＡ含量が１１％であった。得られたリン脂質組成物中のスフィンゴミエリンの構成脂肪酸組成を表２に示す。
Example 3 Preparation of Phospholipid Composition with High Phospholipid Content 2 g of the phospholipid composition obtained in Example 1 was dissolved in chloroform and filled with 200 g of silica gel (Wakogel C-200, manufactured by Wako Pure Chemical Industries, Ltd.). Purification was performed by silica column chromatography on the column, and only the fraction corresponding to phospholipid was separated and collected. After the neutral lipid fraction was washed away with 600 ml of chloroform, the phospholipid fraction was eluted with 1000 ml of methanol, and the solvent was distilled off to obtain 1 g of a phospholipid-rich phospholipid composition. The obtained phospholipid composition had a total phospholipid content of 91%, a sphingomyelin content of 33%, a glycerophospholipid content of 58%, and a DHA content in the glycerophospholipid content of 11%. Table 2 shows the constituent fatty acid composition of sphingomyelin in the obtained phospholipid composition.
実施例４ スフィンゴミエリン高含有リン脂質組成物の調製
実施例１で得たリン脂質組成物４０ｇに０．４Ｎ ＫＯＨ−メタノール溶液１２００ｍｌを加え、４０℃で８時間攪拌し、グリセロリン脂質の加水分解処理を行なった。次にこのものに酢酸２７ｍｌを加え中和し、さらにクロロホルム２４００ｍｌと水１２００ｍｌを加え攪拌後、静置してクロロホルム層を分取した。クロロホルムを留去したものを５℃の冷アセトン２００ｍｌで３回洗浄し、冷アセトン不溶性の沈殿部に残存するアセトンを留去してリン脂質組成物を得た。
次にこのリン脂質組成物２ｇを、１００ｇのシリカゲル（ワコーゲルＣ−２００、和光純薬工業社製）を充填したガラスカラムでのケイ酸カラムクロマトグラフィーにより精製を行い、分画・精製した。溶出には４００ｍｌのクロロホルム、４００ｍｌのクロロホルム−メタノール（１：４）、８００ｍｌのメタノールを用い、メタノール溶出画分を分取し、溶剤を留去しスフィンゴミエリン高含有リン脂質組成物１ｇを得た。得られたリン脂質組成物は、総リン脂質含量９５％であり、スフィンゴミエリンが９０％であった。ＴＬＣで確認したところ、グリセロリン脂質のスポットは見られなかった。得られたリン脂質組成物中のスフィンゴミエリンの構成脂肪酸組成を表２に示す。
Example 4 Preparation of Sphingomyelin-Highly Containing Phospholipid Composition To 40 g of the phospholipid composition obtained in Example 1, 1200 ml of 0.4N KOH-methanol solution was added and stirred at 40 ° C. for 8 hours to hydrolyze glycerophospholipid. Was done. Next, 27 ml of acetic acid was added thereto for neutralization, and further 2400 ml of chloroform and 1200 ml of water were added and stirred, and then allowed to stand to separate a chloroform layer. What distilled chloroform was distilled off was wash | cleaned 3 times with 200 ml of cold acetone of 5 degreeC, acetone remaining in the cold acetone insoluble precipitation part was distilled off, and the phospholipid composition was obtained.
Next, 2 g of this phospholipid composition was purified by silica column chromatography on a glass column packed with 100 g of silica gel (Wakogel C-200, manufactured by Wako Pure Chemical Industries, Ltd.), and fractionated and purified. For elution, 400 ml of chloroform, 400 ml of chloroform-methanol (1: 4), and 800 ml of methanol were used. The methanol-eluted fraction was collected, and the solvent was distilled off to obtain 1 g of a phospholipid composition with a high sphingomyelin content. . The resulting phospholipid composition had a total phospholipid content of 95% and sphingomyelin was 90%. When confirmed by TLC, no spot of glycerophospholipid was observed. Table 2 shows the constituent fatty acid composition of sphingomyelin in the obtained phospholipid composition.
実施例５ グリセロリン脂質高含有リン脂質組成物の調製
実施例１で得たリン脂質組成物２ｇをクロロホルムに溶解し、１００ｇのシリカゲル（ワコーゲルＣ−２００、和光純薬工業社製）を充填したガラスカラムでのケイ酸カラムクロマトグラフィーにより精製を行い、グリセロリン脂質相当画分のみを分離分取した。クロロホルム３００ｍｌで中性脂質画分を洗い流した後、３００ｍｌのクロロホルム−メタノール（２：１）、メタノール１０００ｍｌでリン脂質を溶出させ各２０ｍｌずつの溶出画分として分取し、各画分の溶剤を留去した。ＴＬＣで各画分に含まれるリン脂質種を確認し、グリセロリン脂質を主体とする画分を集め、グリセロリン脂質高含有リン脂質組成物０．４ｇを得た。得られたリン脂質組成物は、総リン脂質含量８５．３％であり、スフィンゴミエリンを１６．４％、グリセロリン脂質を６８．９％含有していた。グリセロリン脂質中のＤＨＡ含量は１１．６％であった。また、グリセロリン脂質中のホスファチジルコリンは７２．１％、リゾホスファチジルコリンは２６．３％で、合計で９８．４％であるのに対しホスファチジルエタノールアミンはわずか１．６％であり、その他のグリセロリン脂質は検出されなかった。得られたリン脂質組成物中のグリセロリン脂質の構成脂肪酸組成を表１に示す。
Example 5 Preparation of phospholipid composition with a high content of glycerophospholipid 2 g of the phospholipid composition obtained in Example 1 was dissolved in chloroform and filled with 100 g of silica gel (Wakogel C-200, manufactured by Wako Pure Chemical Industries, Ltd.). Purification was performed by silica column chromatography on the column, and only the fraction corresponding to glycerophospholipid was separated and collected. After washing away the neutral lipid fraction with 300 ml of chloroform, the phospholipid was eluted with 300 ml of chloroform-methanol (2: 1) and 1000 ml of methanol, and each 20 ml fraction was taken out, and the solvent of each fraction was removed. Distilled off. The phospholipid species contained in each fraction were confirmed by TLC, and fractions mainly composed of glycerophospholipid were collected to obtain 0.4 g of a glycerophospholipid-rich phospholipid composition. The resulting phospholipid composition had a total phospholipid content of 85.3%, containing 16.4% sphingomyelin and 68.9% glycerophospholipid. The DHA content in the glycerophospholipid was 11.6%. In addition, phosphatidylcholine in glycerophospholipid is 72.1%, lysophosphatidylcholine is 26.3%, the total is 98.4%, while phosphatidylethanolamine is only 1.6%, and other glycerophospholipids are Not detected. Table 1 shows the constituent fatty acid composition of glycerophospholipid in the obtained phospholipid composition.
実施例６ ＤＡＧ添加分離
実施例１で得た魚原油処理物の中間層に、ジアシルグリセロール（ＤＡＧ）高含有油脂を添加することで、リン脂質を高純度に含む画分を回収した。
ＤＡＧ高含有油脂は、なたね油、水酸化ナトリウム、グリセリン、水を真空条件下で撹拌・加温・反応させ製造した。ＤＡＧ高含有油脂中のＤＡＧ含量は約４０％であった。実施例１で得た魚原油処理物の中間層３００ｇに対してＤＡＧ高含有油脂を９ｇ（ＤＡＧとして１．２％量）添加し、よく混合した後５０℃で３時間静置した。静置後、遠心分離機にかけ、上層（油）・中間層（リン脂質）・下層（水）の３層に分けた中間層を回収した。
得られた中間層は１３０ｇ、中間層の水分は５０％であり、これを実施例１と同様にアセトン処理して、リン脂質組成物２２ｇを得た。得られたリン脂質組成物は、総リン脂質含量４９％であり、スフィンゴミエリンを１４．５％、グリセロリン脂質を３４．５％含有し、グリセロリン脂質中のＤＨＡ含量が１０．４％であった。これにより、実施例１より高純度のリン脂質組成物を得ることができた。
Example 6 DAG addition separation The fraction which contains phospholipid with high purity was collect | recovered by adding diacylglycerol (DAG) high content fat to the intermediate | middle layer of the processed fish crude oil obtained in Example 1. FIG.
The DAG-rich oil and fat was produced by stirring, heating, and reacting rapeseed oil, sodium hydroxide, glycerin, and water under vacuum conditions. The DAG content in the DAG-rich oil and fat was about 40%. 9 g (1.2% amount of DAG) of high DAG content was added to 300 g of the intermediate layer of processed fish crude oil obtained in Example 1, and after mixing well, the mixture was allowed to stand at 50 ° C. for 3 hours. After allowing to stand, the mixture was centrifuged to collect an intermediate layer divided into three layers of an upper layer (oil), an intermediate layer (phospholipid), and a lower layer (water).
The obtained intermediate layer was 130 g, and the water content of the intermediate layer was 50%. This was treated with acetone in the same manner as in Example 1 to obtain 22 g of a phospholipid composition. The obtained phospholipid composition had a total phospholipid content of 49%, 14.5% sphingomyelin, 34.5% glycerophospholipid, and a DHA content in glycerophospholipid of 10.4%. . Thereby, a higher purity phospholipid composition than Example 1 could be obtained.
実施例７ リン脂質組成物を含む飼料
実施例１で得られたリン脂質組成物２０ｇを細かく切断し、２０ｍｌの水を加え撹拌した。このリン脂質溶液に魚粉４２ｇ、カゼイン６０ｇ、コーングルテン１６ｇ、セルロース１５ｇ、ビタミン１０ｇ、ミネラル１６ｇ、タウリン１ｇを加え、固まりが出来ないようによく混合した。この混合物に水６０ｍｌ、フィードオイル２０ｇ（植田製油製）を少しずつ加え、手でよく捏ね合わせた後、−８０℃冷凍庫で凍結させた。凍結乾燥機により凍結した飼料の水分を取り除いた後、ステンレスのふるいにかけ直径１ｍｍ程度の粒型に造粒したものを試験用飼料とした。
Example 7 Feed Containing Phospholipid Composition 20 g of the phospholipid composition obtained in Example 1 was finely cut, and 20 ml of water was added and stirred. To this phospholipid solution, 42 g of fish meal, 60 g of casein, 16 g of corn gluten, 15 g of cellulose, 10 g of vitamins, 16 g of minerals, and 1 g of taurine were added and mixed well so that no lumps were formed. To this mixture, 60 ml of water and 20 g of feed oil (manufactured by Ueda Oil Co., Ltd.) were added little by little, kneaded well by hand, and then frozen in a −80 ° C. freezer. After removing water from the frozen feed with a freeze dryer, the feed was granulated into a granule having a diameter of about 1 mm through a stainless steel sieve and used as a test feed.
比較例３
魚粉４２ｇ、カゼイン６０ｇ、コーングルテン１６ｇ、セルロース１５ｇ、ビタミン１０ｇ、ミネラル１６ｇ、タウリン１ｇを固まりが出来ないようによく混合した。この混合物に水６０ｍｌ、フィードオイル２０ｇ（植田製油製）、パーム油２０gを少しずつ加え、手でよく捏ね合わせた後、−８０℃冷凍庫で凍結させた。凍結乾燥機により凍結した飼料の水分を取り除いた後、ステンレスのふるいにかけ直径１ｍｍ程度の粒型に造粒したものを試験用飼料とした。
Comparative Example 3
42 g of fish meal, 60 g of casein, 16 g of corn gluten, 15 g of cellulose, 10 g of vitamins, 16 g of minerals, and 1 g of taurine were mixed well so as not to clump. To this mixture, 60 ml of water, 20 g of feed oil (manufactured by Ueda Oil Co., Ltd.) and 20 g of palm oil were added little by little, kneaded well by hand, and then frozen in a −80 ° C. freezer. After removing water from the frozen feed with a freeze dryer, the feed was granulated into a granule having a diameter of about 1 mm through a stainless steel sieve and used as a test feed.
マダイ稚魚摂餌促進効果比較試験
直径７０ｃｍ、深さ８０ｃｍの２００Ｌサイズ円柱型水槽に、生後４ヶ月・体長７ｃｍ程度のマダイ稚魚を４０尾用意した。喰い付きの比較試験は、２種類のエサを水槽左右の水面にそれぞれ０．１ｇずつ同時に投入し、集まるマダイ稚魚の個体数で判断した。試験は１日１回、３日間に渡り反復して行った。
Comparison test of feeding effect of young red sea bream forty 40 young red sea bream with a diameter of 70 cm and a depth of 80 cm and a length of about 7 cm were prepared for 4 months after birth. In the comparative test with bite, two kinds of food were simultaneously added to the water surfaces on the left and right sides of the water tank, respectively, and judged by the number of collected red sea bream fry. The test was repeated once a day for 3 days.
実施例７の飼料に集まるマダイはおよそ１５尾前後であったが、比較例３の飼料に集まるマダイはおよそ７尾前後であった。また、同時に投入した２種の飼料が全て食べ尽くされるまでの時間は、実施例７の飼料がおよそ５秒、比較例３の飼料がおよそ１２秒であった。本発明のリン脂質を含有する飼料は摂餌促進効果があった。 The red sea bream gathered in the feed of Example 7 was about 15 fish, while the red sea bream gathered in the feed of Comparative Example 3 was around 7 fish. Moreover, the time until all of the two kinds of feeds simultaneously fed were consumed was about 5 seconds for the feed of Example 7 and about 12 seconds for the feed of Comparative Example 3. The feed containing the phospholipid of the present invention had a feeding promotion effect.
以上記載したごとく、本発明は複雑な工程を必要とせずにスフィンゴミエリン、及びＤＨＡが結合したグリセロリン脂質を共に含有するリン脂質組成物を提供するものであり、それぞれの含量や結合脂肪酸の特徴による機能を解明する研究や期待される効果を活かした用途として飼料、食品、化粧品、試薬、医薬品等の分野で利用可能である。 As described above, the present invention provides a phospholipid composition containing both sphingomyelin and glycerophospholipid bound to DHA without requiring a complicated process, depending on the content and characteristics of the bound fatty acid. It can be used in the fields of feed, food, cosmetics, reagents, pharmaceuticals, etc. as research that elucidates the function and uses the expected effects.
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| WO2021132516A1 (en) * | 2019-12-26 | 2021-07-01 | マルハニチロ株式会社 | Roe lipid composition containing polyvalent unsaturated fatty acid-bound phospholipid |
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