JPH0439398A - Method for fractionating and purifying lipid fraction from hydrophilic organic solvent extract of animal fats and oils - Google Patents
Method for fractionating and purifying lipid fraction from hydrophilic organic solvent extract of animal fats and oilsInfo
- Publication number
- JPH0439398A JPH0439398A JP2146921A JP14692190A JPH0439398A JP H0439398 A JPH0439398 A JP H0439398A JP 2146921 A JP2146921 A JP 2146921A JP 14692190 A JP14692190 A JP 14692190A JP H0439398 A JPH0439398 A JP H0439398A
- Authority
- JP
- Japan
- Prior art keywords
- organic solvent
- fraction
- hydrophilic organic
- fractionating
- oils
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000003921 oil Substances 0.000 title claims abstract description 28
- 239000003960 organic solvent Substances 0.000 title claims abstract description 28
- 239000003925 fat Substances 0.000 title claims abstract description 25
- 239000000284 extract Substances 0.000 title claims abstract description 24
- 241001465754 Metazoa Species 0.000 title claims abstract description 22
- 150000002632 lipids Chemical class 0.000 title claims abstract description 17
- 238000000034 method Methods 0.000 title claims description 25
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract description 46
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 31
- 235000012000 cholesterol Nutrition 0.000 claims abstract description 23
- 239000002904 solvent Substances 0.000 claims abstract description 23
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 23
- 150000003904 phospholipids Chemical class 0.000 claims abstract description 20
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 19
- 229930195734 saturated hydrocarbon Natural products 0.000 claims abstract description 16
- 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 abstract description 13
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 claims abstract description 13
- 239000012046 mixed solvent Substances 0.000 claims abstract description 12
- 238000000622 liquid--liquid extraction Methods 0.000 claims abstract description 11
- 238000004587 chromatography analysis Methods 0.000 claims abstract description 9
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 claims abstract description 7
- 150000008104 phosphatidylethanolamines Chemical class 0.000 claims abstract description 7
- 235000019197 fats Nutrition 0.000 claims description 22
- 235000019737 Animal fat Nutrition 0.000 claims description 6
- 235000014113 dietary fatty acids Nutrition 0.000 claims description 5
- 239000000194 fatty acid Substances 0.000 claims description 5
- 229930195729 fatty acid Natural products 0.000 claims description 5
- -1 fatty acid triglycerides Chemical class 0.000 claims description 4
- 150000004665 fatty acids Chemical class 0.000 claims description 4
- 238000000746 purification Methods 0.000 claims description 3
- 239000010775 animal oil Substances 0.000 claims 1
- 238000005194 fractionation Methods 0.000 claims 1
- 150000002270 gangliosides Chemical class 0.000 abstract description 4
- WPIHMWBQRSAMDE-YCZTVTEBSA-N beta-D-galactosyl-(1->4)-beta-D-galactosyl-N-(pentacosanoyl)sphingosine Chemical compound CCCCCCCCCCCCCCCCCCCCCCCCC(=O)N[C@@H](CO[C@@H]1O[C@H](CO)[C@H](O[C@@H]2O[C@H](CO)[C@H](O)[C@H](O)[C@H]2O)[C@H](O)[C@H]1O)[C@H](O)\C=C\CCCCCCCCCCCCC WPIHMWBQRSAMDE-YCZTVTEBSA-N 0.000 abstract description 3
- 150000004666 short chain fatty acids Chemical class 0.000 abstract description 3
- 150000003431 steroids Chemical class 0.000 abstract description 3
- 150000002305 glucosylceramides Chemical class 0.000 abstract description 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 27
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 27
- 235000019198 oils Nutrition 0.000 description 19
- 238000010828 elution Methods 0.000 description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- 230000005526 G1 to G0 transition Effects 0.000 description 9
- 238000000605 extraction Methods 0.000 description 8
- 239000007788 liquid Substances 0.000 description 7
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 description 6
- 230000001174 ascending effect Effects 0.000 description 6
- 235000014121 butter Nutrition 0.000 description 5
- 235000013305 food Nutrition 0.000 description 4
- 229940106189 ceramide Drugs 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000002844 melting Methods 0.000 description 3
- 230000008018 melting Effects 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 150000003626 triacylglycerols Chemical class 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- YDNKGFDKKRUKPY-JHOUSYSJSA-N C16 ceramide Natural products CCCCCCCCCCCCCCCC(=O)N[C@@H](CO)[C@H](O)C=CCCCCCCCCCCCCC YDNKGFDKKRUKPY-JHOUSYSJSA-N 0.000 description 2
- 229930186217 Glycolipid Natural products 0.000 description 2
- CRJGESKKUOMBCT-VQTJNVASSA-N N-acetylsphinganine Chemical compound CCCCCCCCCCCCCCC[C@@H](O)[C@H](CO)NC(C)=O CRJGESKKUOMBCT-VQTJNVASSA-N 0.000 description 2
- OFBQJSOFQDEBGM-UHFFFAOYSA-N Pentane Chemical compound CCCCC OFBQJSOFQDEBGM-UHFFFAOYSA-N 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- ZVEQCJWYRWKARO-UHFFFAOYSA-N ceramide Natural products CCCCCCCCCCCCCCC(O)C(=O)NC(CO)C(O)C=CCCC=C(C)CCCCCCCCC ZVEQCJWYRWKARO-UHFFFAOYSA-N 0.000 description 2
- 235000021323 fish oil Nutrition 0.000 description 2
- 229930182478 glucoside Natural products 0.000 description 2
- 150000008131 glucosides Chemical class 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- VVGIYYKRAMHVLU-UHFFFAOYSA-N newbouldiamide Natural products CCCCCCCCCCCCCCCCCCCC(O)C(O)C(O)C(CO)NC(=O)CCCCCCCCCCCCCCCCC VVGIYYKRAMHVLU-UHFFFAOYSA-N 0.000 description 2
- 235000014593 oils and fats Nutrition 0.000 description 2
- 230000001766 physiological effect Effects 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000000638 solvent extraction Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000013076 target substance Substances 0.000 description 2
- 206010003210 Arteriosclerosis Diseases 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- 239000004809 Teflon Substances 0.000 description 1
- 229920006362 Teflon® Polymers 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 150000001783 ceramides Chemical class 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000006694 eating habits Nutrition 0.000 description 1
- 235000013345 egg yolk Nutrition 0.000 description 1
- 210000002969 egg yolk Anatomy 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000004146 energy storage Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 150000002170 ethers Chemical class 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 150000002314 glycerols Chemical class 0.000 description 1
- 150000002327 glycerophospholipids Chemical class 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 238000005191 phase separation Methods 0.000 description 1
- 235000015277 pork Nutrition 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 235000003441 saturated fatty acids Nutrition 0.000 description 1
- 150000004671 saturated fatty acids Chemical class 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000003760 tallow Substances 0.000 description 1
Landscapes
- Fats And Perfumes (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明は動物性油脂を親水性有機溶媒で処理して抽出し
た抽出物中に含まれる各脂質画分をそれぞれ効率よく、
高純度に分画精製する方法に関する。[Detailed Description of the Invention] [Industrial Application Field] The present invention efficiently extracts each lipid fraction contained in an extract obtained by treating animal fats and oils with a hydrophilic organic solvent.
This invention relates to a method for fractionating and purifying to high purity.
中性脂質は脂肪酸のグリセリンエステルである。 Neutral lipids are glycerol esters of fatty acids.
動物の主な中性脂質はステアリン酸、バルミチン酸のよ
うな飽和脂肪酸を多く含むトリアジルグリセロール（ト
リグリセリドともいう）で、常温で固体のことが多い。The main neutral lipid in animals is triadylglycerol (also called triglyceride), which contains a lot of saturated fatty acids such as stearic acid and balmitic acid, and is often solid at room temperature.
生体の有効なエネルギー貯蔵物質であり、腸管リパーゼ
で加水分解され吸収される。細胞膜で代謝的に生ずるア
シルグリセロールのうちには細胞におけるホルモン作用
など情報伝達の仕組みの上で重要な役割を演じているも
のがある。It is an effective energy storage substance for living organisms and is hydrolyzed and absorbed by intestinal lipase. Among the acylglycerols that are metabolically produced in cell membranes, some play an important role in information transmission mechanisms such as hormone effects in cells.
コレステロールはステロイドの一種で、生体内で生産さ
れ、溶血物に対して赤血球を保護する作用がある。Cholesterol is a type of steroid that is produced within the body and has the effect of protecting red blood cells against hemolysates.
天然にはグリセロリン脂質に属するフォスファチジルコ
リン（ＰＣ）、フォスファチジルエタノールアミン（Ｐ
Ｅ）やスフィンゴリン脂質に属するスフィンゴミエリン
（ＳＰＭ）などのリン脂質が存在する。Naturally, phosphatidylcholine (PC) and phosphatidylethanolamine (P), which belong to glycerophospholipids, are
There are phospholipids such as E) and sphingomyelin (SPM), which belongs to sphingophospholipids.
ＰＣは天然に最も広く分布しているリン脂質であって、
界面活性作用があるので乳化剤として食品添加物など、
或いは医薬、その他の工業用原料として広く利用されて
いる。また、ＰＥ、 ＳＰＭなども生理活性を有するこ
とが知られている。特にＳＰＭはＰＡＦ（Ｐｌａｔｅｌ
ｅＬ Ａｃｔｉｖａｔｉｎｇ Ｆａｃｔｏｒ） の血小
牟反凝集に対するアンタゴニストとして注目され始めて
いる［Ｊ、Ｂｉｏｌ、Ｃｈｅｍ、 、２６２．１３１７
４（１９８７） ）。PC is the most widely distributed phospholipid in nature,
Due to its surface-active effect, it can be used as an emulsifier in food additives, etc.
Alternatively, it is widely used as a raw material for medicine and other industrial purposes. Furthermore, PE, SPM, and the like are also known to have physiological activity. In particular, SPM is PAF (Platel
eL Activating Factor) is beginning to attract attention as an antagonist against blood cell aggregation [J, Biol, Chem, 262.1317
4 (1987)).
これらのリン脂質は、主に卵黄、大豆などから溶剤で抽
出し、これを分画精製することによって製造されていた
。また、リン脂質の一部は化学的にも合成されていた。These phospholipids were mainly produced by extracting them from egg yolks, soybeans, etc. with solvents and fractionating and purifying them. In addition, some phospholipids were also synthesized chemically.
糖脂質としては、種々の生理活性をもつラクトシルセラ
ミド、グルコシドセラミド、ガングリオシドなどがある
。Glycolipids include lactosylceramide, glucoside ceramide, and ganglioside, which have various physiological activities.
一方、食生活の洋風化に伴って、肉類やバターなどの動
物性脂肪を含む食品を多量に摂取すると動物性油脂中に
含まれるコレステロールによって動脈硬化、心筋梗塞、
脳卒中などを発症するという問題が発生し、コレステロ
ール含量を低減した動物性油脂を含む食品が要望される
ようになってきている現状にある。On the other hand, as dietary habits become more Westernized, if we consume large amounts of foods containing animal fats such as meat and butter, the cholesterol contained in the animal fats and oils may cause arteriosclerosis, myocardial infarction, and so on.
Due to problems such as the onset of stroke, there is a growing demand for foods containing animal fats and oils with reduced cholesterol content.
そこで、動物性油脂中のコレステロール含量を低減する
有効な方法として、観水性有ｌａｄ媒を用い、動物性油
脂からコレステロールを抽出、除去する方法が提案され
ている（特願平２−２５８１１号）。Therefore, as an effective method for reducing the cholesterol content in animal fats and oils, a method has been proposed in which cholesterol is extracted and removed from animal fats and oils using a hydrophilic rad medium (Japanese Patent Application No. 2-25811). .
この方法は主にエタノールを溶媒として用いるので、安
価で、かつ安全性の高い方法と考えられる。Since this method mainly uses ethanol as a solvent, it is considered to be an inexpensive and highly safe method.
しかしながら、大量に使用した抽出溶媒を蒸留して回収
する程度で、抽出液中に含まれている各種成分の回収に
ついては、全く考慮されていない現状にあった。However, the current situation is that the extraction solvent used in large quantities is recovered by distillation, and no consideration is given to the recovery of various components contained in the extract.
〔発明が解決しようとするｉＩ題）
本発明者らは、上述した状況に鑑み、動物性油脂からコ
レステロールを抽出、除去するのに用いた親水性有Ｎ
溶媒から各種成分を回収し、利用することを目的として
鋭意研究を重ねてきた結果、最近、微量成分を分離する
為に開発された遠心液々分配クロマトグラフ（Ｃｅｎｔ
ｒｉｆｕｇａｌ Ｐａｒｔｉ ＬｉｏｎＣｈｒｏ＋ｍａ
ｔｏｇｒａｐｈ） （以下、ｃｐｃという）を用い、適
当な溶媒を選択することによって、抽出液中に含まれる
フォスファチジルコリン、フォスファチジルエタノール
アミン、スフィンゴミエリンなどのリン脂質画分、更に
はラクトシルセラミド、グルコシルセラミド、ガングリ
オシドなどのグリセロ糖脂質画分、コレステロールなど
のステロイド画分、短鎖脂肪酸トリグリセリド画分を効
率よく分画精製出来るとの知見を得、本発明をなすに至
、った。[Problem to be Solved by the Invention] In view of the above-mentioned situation, the present inventors have developed a hydrophilic nitrogen-containing compound used to extract and remove cholesterol from animal fats and oils.
As a result of extensive research aimed at recovering and utilizing various components from solvents, we have recently developed a centrifugal liquid-liquid partition chromatograph (Centrifugal Liquid Partition Chromatograph) to separate trace components.
rifugal Party LionChro+ma
phospholipid fractions such as phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin contained in the extract, as well as lactosyl We have found that glyceroglycolipid fractions such as ceramides, glucosylceramides, and gangliosides, steroid fractions such as cholesterol, and short-chain fatty acid triglyceride fractions can be efficiently fractionated and purified, leading to the present invention.
従って本発明は、動物性油脂からコレステロールを抽出
、除去するのに用いた親水性有機溶媒から各脂質画分を
分画精製する方法を提供することを課題とする。Therefore, an object of the present invention is to provide a method for fractionating and purifying each lipid fraction from a hydrophilic organic solvent used to extract and remove cholesterol from animal fats and oils.
本発明の脂質画分の分画精製法は、動物性油脂を親水性
有機溶媒で処理して抽出した抽出物を特定の溶媒を用い
て遠心液々分配クロマトグラフにかけて各脂質画分を分
画、精製する方法に関する。The method for fractionating and purifying lipid fractions of the present invention involves treating animal fats and oils with a hydrophilic organic solvent and subjecting the extract to centrifugal liquid-liquid partition chromatography using a specific solvent to separate each lipid fraction. , relating to a method of purification.
本発明における動物性油脂には、コレステロールを含む
バター、バターからクリームを分離したバターオイル、
ラード、牛脂、豚脂、羊脂、魚油、肝油などがある。ま
た、親水性有機溶媒には、エタノール、アセトン等を用
いることができるが、食品衛生上からはエタノールを用
いることが好ましい。これらの親水性有機溶媒は含水の
状態であってもよい。しかし含水率が１０容量％以上の
ものを用いることが好ましい。含水率が１０容量％以↓
２のものを使用すると脂質の回収率が低下するので好ま
しくない。抽出手段としては、回分式の抽出装置、連続
式多段向流抽出装置等による液−液抽出方式が好ましい
。In the present invention, animal fats and oils include butter containing cholesterol, butter oil obtained by separating cream from butter,
Examples include lard, beef tallow, pork fat, mutton fat, fish oil, and liver oil. Further, ethanol, acetone, etc. can be used as the hydrophilic organic solvent, but from the viewpoint of food hygiene, it is preferable to use ethanol. These hydrophilic organic solvents may be in a water-containing state. However, it is preferable to use one having a water content of 10% by volume or more. Moisture content is 10% by volume or more↓
Use of No. 2 is not preferable because the lipid recovery rate decreases. As the extraction means, a liquid-liquid extraction method using a batch type extraction device, a continuous multi-stage countercurrent extraction device, etc. is preferable.
動物性油脂を、親水性有機溶媒で抽出するには、動物性
油脂を油脂の融点以上の温度で約１０分〜１時間程度接
触させた後、同温度の下に約２０分〜１時間程度静置し
て相分離させる。なお、動物性油脂の融点は、種類によ
り異なるが、一般に常温で固体を呈する油脂では３０〜
５０°Ｃで抽出し、魚油のような常温で液状を呈する油
脂では２０〜３０’Ｃで抽出することが好ましい。この
相分離によって得られる上層を親水性有ｌｌ溶媒抽出物
とする。To extract animal fats and oils with a hydrophilic organic solvent, contact the animal fats and oils at a temperature above the melting point of the fat for about 10 minutes to 1 hour, and then at the same temperature for about 20 minutes to 1 hour. Let stand to phase separate. The melting point of animal fats and oils varies depending on the type, but in general oils and fats that are solid at room temperature have a melting point of 30~30.
It is preferable to extract at 50°C, and for oils and fats that are liquid at room temperature, such as fish oil, to be extracted at 20 to 30'C. The upper layer obtained by this phase separation is used as a hydrophilic solvent extract.
本発明では、このような親水性有機溶媒抽出物を遠心液
々分配クロマトグラフ（ＣＰＣ）で処理する。In the present invention, such a hydrophilic organic solvent extract is processed by centrifugal liquid-liquid partition chromatography (CPC).
ＣＰＣ処理は、例えば特開昭５９−６２３１２号公報に
記載されているように、二層に分離される二層分離液を
溶媒として使用し、そのうち一方を固定相として遠心力
により保持しつつ、他方を移動相として連続的に固定相
内を通過させて移動相内に注入された試料を分画精製す
る方法である。As described in, for example, Japanese Unexamined Patent Publication No. 59-62312, the CPC treatment uses a two-layer separated liquid as a solvent, one of which is used as a stationary phase and is held by centrifugal force. In this method, the sample injected into the mobile phase is fractionated and purified by continuously passing the other phase through the stationary phase.
本発明における親水性有機溶媒抽出物には、中性脂質、
リン脂質及びグリセロ糖脂質が含まれており、本発明で
は、これらの両分を溶媒に対する溶解度の差を利用して
ｃｐｃ処理により分画する。The hydrophilic organic solvent extract in the present invention includes neutral lipids,
It contains phospholipids and glyceroglycolipids, and in the present invention, these two components are fractionated by CPC treatment using the difference in solubility in solvents.
ｃｐｃ処理に使用する溶媒は、通常混合溶媒が使用され
る。混合溶媒は、リン脂質、グリセロ糖脂質の分画精製
をすることが出来、安価で食品工業などに異和感のない
ものから選定される。このような観点から、飽和炭化水
素、アルコール、水よりなる混合溶媒、或いは飽和炭化
水素、エーテル、アルコール、水よりなる混合溶媒が用
いられる。The solvent used for CPC treatment is usually a mixed solvent. The mixed solvent is selected from among those that can perform fractional purification of phospholipids and glyceroglycolipids, are inexpensive, and are compatible with the food industry. From this point of view, a mixed solvent consisting of a saturated hydrocarbon, alcohol, and water, or a mixed solvent consisting of a saturated hydrocarbon, ether, alcohol, and water is used.
飽和炭化水素では、ペンタン、ヘキサン、ヘプタンなど
が望ましく、エーテルとしては、ジエチルエーテルが望
ましく、アルコールとしては、メタノール、エタノール
が望ましい。Preferred saturated hydrocarbons include pentane, hexane, and heptane; preferred ethers are diethyl ether; and preferred alcohols are methanol and ethanol.
またｃｐｃ処理は、下層液を固定相、上層液を移動相と
して用いる上昇法、１層液を固定相、下層液を移動相と
して用いる下降法のどちらかの分離方法を選択して行わ
れる。更に、溶出は目的物を移動相に分配させる正溶出
、もしくは固定相に分配させる反転溶出のどちらか選択
して行われる。The CPC treatment is carried out by selecting one of the following separation methods: an ascending method in which the lower layer liquid is used as a stationary phase and an upper layer liquid as a mobile phase, and a descending method in which the first layer liquid is used as a stationary phase and the lower layer liquid is used as a mobile phase. Further, elution is performed by selecting either forward elution in which the target substance is distributed into the mobile phase or reverse elution in which the target substance is distributed into the stationary phase.
本発明の動物性油脂由来の脂質画分の分画、精製方法の
概略を第１図を示しながら説明する。The outline of the method for fractionating and purifying lipid fractions derived from animal fats and oils of the present invention will be explained with reference to FIG.
まず、動物性油脂を、固体の場合は、加’ＩＡＬ、で溶
融し、これに親水性有機溶媒を加えて撹拌し、抽出を行
う。次に、これを遠心分離して上層の親水性有機溶媒層
を採取し、この撹拌抽出及び遠心分離の操作を数回繰り
返し親水性有機溶媒層を集め、減圧下に濃縮乾固して親
水性有４！！ａ溶媒抽出物を得る。First, if the animal fat or oil is solid, it is melted using an aliquot, and a hydrophilic organic solvent is added thereto and stirred to perform extraction. Next, this is centrifuged to collect the upper hydrophilic organic solvent layer, this operation of stirring extraction and centrifugation is repeated several times to collect the hydrophilic organic solvent layer, and the hydrophilic organic solvent layer is concentrated to dryness under reduced pressure. Yes 4! ! a Obtain a solvent extract.
この親水性有機溶媒抽出物を飽和炭化水素、アルコール
及び水よりなる溶媒、例えばヘプタン、メタノール及び
水よりなる溶媒を使用して上昇法でＣＰＣ処理を行い、
トリグリセリド・コレステロ−ル画分、リン脂質画分！
、リン脂質画分■及びグリセロ糖脂質画分に分画する。This hydrophilic organic solvent extract is subjected to CPC treatment in an ascending method using a solvent consisting of a saturated hydrocarbon, alcohol and water, such as heptane, methanol and water,
Triglyceride/cholesterol fraction, phospholipid fraction!
, a phospholipid fraction (■) and a glyceroglycolipid fraction.
ここで得られるトリグリセリド・コレステロール画分は
、飽和炭化水素、アルコール及び水よりなる溶媒、例え
ばヘキサン、メタノール及び水よりなる溶媒を使用して
上昇法でＣＰＣ処理を行いトリグリセリドとコレステロ
ールとに分画し、高純度のコレステロールを採取し、ま
たトリグリセリドは、飽和炭化水素、アルコール及び酢
酸よりなる溶媒、例えばヘキサン、メタノール及び酢酸
よりなる溶媒を用い上昇法でｃｐｃ処理を行って短鎖脂
肪酸トリグリセリドを得る。The triglyceride/cholesterol fraction obtained here is fractionated into triglycerides and cholesterol by CPC treatment using an ascending method using a solvent consisting of a saturated hydrocarbon, alcohol, and water, such as hexane, methanol, and water. Highly purified cholesterol is collected, and triglyceride is subjected to CPC treatment using a solvent consisting of a saturated hydrocarbon, alcohol, and acetic acid, such as hexane, methanol, and acetic acid, using an ascending method to obtain short chain fatty acid triglycerides.
一方、リン脂質画分Ｉは、飽和炭化水素、アルコール及
び水よりなる溶媒、例えばヘキサン、エタノール及び水
よりなる溶媒を用い上昇法でＣＰＣ処理を行って高純度
のＰＥ画分を採取する。On the other hand, for the phospholipid fraction I, a highly pure PE fraction is collected by CPC treatment using an ascending method using a solvent consisting of a saturated hydrocarbon, alcohol, and water, such as hexane, ethanol, and water.
また、ＰＣ，ＳＰＭの混合画分であるリン脂質画分■は
、飽和炭化水素、エーテル、アルコール及び水よりなる
溶媒、例えばヘキサン、ジエチルエーテル、メタノール
及び水よりなる溶媒を用い上昇法でｃｐｃ処理を行って
ｐｃ画分とＳＰＭ画分とに分画し、それぞれの両分を採
取する。In addition, the phospholipid fraction (2), which is a mixed fraction of PC and SPM, is subjected to CPC treatment using an ascending method using a solvent consisting of saturated hydrocarbon, ether, alcohol, and water, such as hexane, diethyl ether, methanol, and water. The sample is fractionated into a PC fraction and an SPM fraction, and both fractions are collected.
本発明を実施例を挙げて具体的に説明する。The present invention will be specifically described with reference to Examples.
実施例１
バター１５０ｇを加温し、融解したものに４５０ｄのエ
タノールを加え、長さ４０閣のテフロン製撹拌子を用い
て、５０°Ｃで２０分間、毎分５００回転で撹拌し、抽
出を行った後、毎分３０００回転、１５分間の条件で遠
心分離し、上層のエタノール層を採集し、更に同一の条
件で２回抽出を繰り返した後、親水性有機溶媒抽出物１
３５０ｄを得た。これを減圧下で濃縮、乾固して親水性
有機溶媒抽出物１１．２ｇを得た。Example 1 Heat 150 g of butter, add 450 d of ethanol to the melted mixture, and stir at 500 rpm for 20 minutes at 50°C using a Teflon stirrer with a length of 40 mm to perform extraction. After that, centrifugation was performed at 3,000 revolutions per minute for 15 minutes, the upper ethanol layer was collected, and the extraction was repeated twice under the same conditions, followed by hydrophilic organic solvent extract 1.
I got 350d. This was concentrated and dried under reduced pressure to obtain 11.2 g of a hydrophilic organic solvent extract.
この親水性有機溶媒抽出！１ｈ１１．２ｇを、ヘプタン
：メタノール：水＝２００：９７：３の比率にし、その
下層を固定相とし、上層を移動相とした溶媒系を用いｃ
ｐｃに供給した。この時の運転条件は、２０°Ｃ１流速
１５．０ｄ／＋ｉｎ、容量１０，０００Ｍ１、回転数１
．６０Ｏｒ、ｐ、ｍであった。この操作によってこの両
分を正溶出でトリグリセリド・コレステロール画分１０
．７５ ｇ、リン脂質画分１１２４■、リン脂質画分■
１６３■及びグリセロ糖脂質画分２１■に分画した。This hydrophilic organic solvent extraction! 11.2 g of 1h was made into a ratio of heptane: methanol: water = 200:97:3, using a solvent system in which the lower layer was the stationary phase and the upper layer was the mobile phase.
Supplied to PC. The operating conditions at this time were 20°C, flow rate 15.0d/+in, capacity 10,000M1, and rotation speed 1.
．． It was 60 Or, p, m. By this procedure, both fractions were separated into triglyceride/cholesterol fraction 10 by normal elution.
．． 75 g, phospholipid fraction 1124■, phospholipid fraction■
It was fractionated into 163■ and glyceroglycolipid fractions 21■.
得られたトリグリセリド・コレステロール画分１０．７
５ｇをヘキサン：メタノール：水＝２００：９５：５の
比率にした溶媒を用い、その下層を固定相とし、上層を
移動相とし、ＣＰＣに供給した。この時の運転条件は、
２５’Ｃ１流速１５．Ｏａｆ／ｗｉｎ、流量４．ＯＯＯ
＃Ｉｌ！、回転数１ 、５０Ｏｒｐｍであった。この操
作で正溶出でトリグリセリド１０．３ｇ、反転溶出でコ
レステロール３１２■を得た。Obtained triglyceride/cholesterol fraction 10.7
Using a solvent containing 5 g of hexane:methanol:water in a ratio of 200:95:5, the lower layer was used as a stationary phase and the upper layer was used as a mobile phase, and the mixture was supplied to CPC. The operating conditions at this time are
25'C1 flow rate 15. Oaf/win, flow rate 4. OOO
#Il! The rotation speed was 1 and 50 rpm. Through this operation, 10.3 g of triglyceride was obtained by forward elution, and 312 g of cholesterol was obtained by reverse elution.
このトリグリセリドを、ヘキサン：メタノール：酢酸＝
５０：３０：１０の比率にした溶媒を用い、同様にして
ＣＰＣに供給した。この時の運転条件は、１５°Ｃ１流
速１０．０ｄ／ｗｉｎ、流１ｔｌ、０００ｍＮ、回転数
１．４００ｒｐＩ１１であった。この操作で正溶出で短
鎖脂肪酸（Ｃ４〜Ｃ８）トリグリセリド２．４ｇを得た
。This triglyceride is converted into hexane:methanol:acetic acid=
A 50:30:10 ratio of solvents was used and fed to the CPC in the same manner. The operating conditions at this time were 15° C., flow rate of 10.0 d/win, flow rate of 1 tl, 000 mN, and rotation speed of 1.400 rpm. Through this operation, 2.4 g of short chain fatty acid (C4 to C8) triglyceride was obtained by normal elution.
一方、リン脂質画分■をヘキサン：エタノール：水−２
００：９０：１０の比率にし、その下層を固定相とし、
上層を移動相とした溶媒系を用いｃｐｃに供給した。こ
の時の運転条件は、２０°Ｃ１流速３．０ｄ／ｗｉｎ、
流量５００ｄ、回転数８００ｒｐ−であった、この操作
により、正溶出でＰＨ画分９７１ｇを得た。この画分は
フォスファチジルエタノールアミン８７％及びフォスフ
ァチジルコリン１３％よりなるものであった。On the other hand, the phospholipid fraction ■ was mixed with hexane:ethanol:water-2
The ratio is 00:90:10, and the lower layer is the stationary phase.
A solvent system with the upper layer as the mobile phase was used to supply the CPC. The operating conditions at this time were 20°C, flow rate 3.0d/win,
Through this operation at a flow rate of 500 d and a rotational speed of 800 rpm, 971 g of a PH fraction was obtained by normal elution. This fraction consisted of 87% phosphatidylethanolamine and 13% phosphatidylcholine.
また、リン脂質画分ＩＩをヘキサン：ジエチルエーテル
：メタノール：水＝１００：１００：８７：１３の比率
にし、その下層を固定相とし、上層を移動相とした溶媒
系を用いｃｐｃに供給した。この時の運転条件は、１５
°Ｃで、流速３ 、０ ｔｎＲ／ ｍ ｉ ｎ、流量１
，５００ｍ、回転数９０Ｏｒｐｍであった。この操作に
より、正溶出でフォスファチジルコリン８２■、反転溶
出でスフィンゴミニリンフ１１１ｇを得た。Further, phospholipid fraction II was supplied to CPC using a solvent system in which the ratio of hexane: diethyl ether: methanol: water = 100:100:87:13 was used, and the lower layer was used as a stationary phase and the upper layer was used as a mobile phase. The operating conditions at this time were 15
°C, flow rate 3, 0 tnR/min, flow rate 1
, 500 m, and the rotation speed was 90 rpm. By this operation, 82 g of phosphatidylcholine was obtained by forward elution, and 111 g of sphingominilymph was obtained by reverse elution.
さらに、前記のようにして得られたグリセロ糖脂質画分
をシリカゲルクロマトグラフィーで分析したところ、ラ
クトシルセラミド３６％、グルコシドセラミド２８％及
びガングリオシドＤ３３２％よりなっていた。Furthermore, when the glyceroglycolipid fraction obtained as described above was analyzed by silica gel chromatography, it was found to consist of 36% lactosylceramide, 28% glucoside ceramide, and 332% ganglioside D.
本発明によると、動物性油脂の親水性有機溶媒抽出物か
らリン脂質、糖脂質、コレステロール、トリグリセリド
等生理活性物質を効率よく、高純度に分画精製すること
ができる。According to the present invention, physiologically active substances such as phospholipids, glycolipids, cholesterol, and triglycerides can be efficiently fractionated and purified to high purity from hydrophilic organic solvent extracts of animal fats and oils.
従って動物性油脂からコレステロールを抽出除去に用い
た溶媒中に含まれる各種有効成分を効率よく回収するこ
とができる。Therefore, various active ingredients contained in the solvent used to extract and remove cholesterol from animal fats and oils can be efficiently recovered.
第１図は、本発明における動物性油脂の親水性有機溶媒
抽出物の分画精製方法の概略を示す図である。
出 願 人 雪印乳業株式会社FIG. 1 is a diagram schematically showing a method for fractionating and purifying a hydrophilic organic solvent extract of animal fat and oil in the present invention. Applicant Snow Brand Milk Products Co., Ltd.
Claims (5)
水素、アルコール及び水の混合溶媒あるいは飽和炭化水
素、エーテル、アルコール及び水の混合溶媒を溶媒とし
て用いて遠心液々分配クロマトグラフにかけて脂肪酸ト
リグリセリド、コレステロール、フォスファチジルエタ
ノールアミン、フォスファチジルコリン、スフィンゴミ
エリン、及び／又はグリセロ糖脂質に分画し、これを精
製することを特徴とする動物性油脂の親水性有機溶媒抽
出物から脂質画分を分画精製する方法。(1) A hydrophilic organic solvent extract of animal fat and oil is subjected to centrifugal liquid-liquid partition chromatography using a mixed solvent of saturated hydrocarbon, alcohol and water or a mixed solvent of saturated hydrocarbon, ether, alcohol and water as a solvent. From a hydrophilic organic solvent extract of animal fat and oil, which is characterized by fractionating and purifying fatty acid triglycerides, cholesterol, phosphatidylethanolamine, phosphatidylcholine, sphingomyelin, and/or glyceroglycolipids. A method for fractionating and purifying lipid fractions.
素、アルコール及び水の混合溶媒を用いて遠心液々分配
クロマトグラフにかけて脂肪酸トリグリセリド・コレス
テロール画分、フォスフアチジルエタノールアミンを主
成分とするリン脂質画分 I 、フォスファチジルコリン
及びスフィンゴミエリンを主成分とするリン脂質画分I
I及び／又はグリセロ糖脂質画分とに分画し、これを精
製することを特徴とする動物性油脂の親水性有機溶媒抽
出物から脂質画分を分画精製する方法。(2) The hydrophilic organic solvent extract of animal oil and fat is subjected to centrifugal liquid-liquid partition chromatography using a mixed solvent of saturated hydrocarbon, alcohol, and water, and the main components are fatty acid triglyceride and cholesterol fractions and phosphatidylethanolamine. Phospholipid fraction I containing phosphatidylcholine and sphingomyelin as main components
1. A method for fractionating and purifying a lipid fraction from a hydrophilic organic solvent extract of animal fat and oil, which comprises fractionating into I and/or glyceroglycolipid fractions and purifying these fractions.
ロール画分を飽和炭化水素、アルコール及び水の混合溶
媒を用いて遠心液々分配クロマトグラフにかけて脂肪酸
トリグリセリドとコレステロールとに分画精製すること
を特徴とする動物性油脂の親水性有機溶媒抽出物から脂
質画分を分画精製する方法。(3) The fatty acid triglyceride/cholesterol fraction of claim (2) is subjected to centrifugal liquid-liquid partition chromatography using a mixed solvent of saturated hydrocarbon, alcohol, and water to separate and purify fatty acid triglyceride and cholesterol. A method for fractionating and purifying lipid fractions from hydrophilic organic solvent extracts of animal fats and oils.
、アルコール及び水の混合溶媒を用いて遠心液々分配ク
ロマトグラフにかけてフォスファチジルエタノールアミ
ンを分画精製することを特徴とする動物性油脂の親水性
有機溶媒抽出物から脂質画分を分画精製する方法。(4) The phospholipid fraction I of claim (2) is subjected to centrifugal liquid-liquid partition chromatography using a mixed solvent of saturated hydrocarbon, alcohol, and water to fractionate and purify phosphatidylethanolamine. A method for fractionating and purifying lipid fractions from hydrophilic organic solvent extracts of animal fats and oils.
エーテル、アルコール及び水の混合溶媒を用いて遠心液
々分配クロマトグラフにかけてフォスファチジルコリン
及びスフィンゴミエリンに分画精製することを特徴とす
る動物性油脂の親水性有機溶媒抽出物から脂質画分を分
画精製する方法。(5) The phospholipid fraction II of claim (2) is a saturated hydrocarbon,
A lipid fraction is obtained from a hydrophilic organic solvent extract of animal fat and oil by fractionation and purification into phosphatidylcholine and sphingomyelin by centrifugal liquid-liquid partition chromatography using a mixed solvent of ether, alcohol and water. How to fractionate and purify.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2146921A JPH0439398A (en) | 1990-06-05 | 1990-06-05 | Method for fractionating and purifying lipid fraction from hydrophilic organic solvent extract of animal fats and oils |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2146921A JPH0439398A (en) | 1990-06-05 | 1990-06-05 | Method for fractionating and purifying lipid fraction from hydrophilic organic solvent extract of animal fats and oils |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH0439398A true JPH0439398A (en) | 1992-02-10 |
Family
ID=15418582
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2146921A Pending JPH0439398A (en) | 1990-06-05 | 1990-06-05 | Method for fractionating and purifying lipid fraction from hydrophilic organic solvent extract of animal fats and oils |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0439398A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2710523A1 (en) * | 1993-09-28 | 1995-04-07 | Ard | Extract of wheat lipids and process for manufacturing it |
| FR2750133A1 (en) * | 1996-06-21 | 1997-12-26 | Ifremer | Purification of phospholipid(s) with high content of poly:unsaturated fatty acids |
| WO2009154309A1 (en) * | 2008-06-20 | 2009-12-23 | 有限会社梅田事務所 | Method for production of highly pure phospholipid, and highly pure sphingomyelin and plasmalogen-type glycerophospholipid produced by the method |
| JP2012170441A (en) * | 2011-02-24 | 2012-09-10 | Ueda Oils & Fats Mfg Co Ltd | Fish-derived phospholipid composition and method for producing the same |
-
1990
- 1990-06-05 JP JP2146921A patent/JPH0439398A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2710523A1 (en) * | 1993-09-28 | 1995-04-07 | Ard | Extract of wheat lipids and process for manufacturing it |
| FR2750133A1 (en) * | 1996-06-21 | 1997-12-26 | Ifremer | Purification of phospholipid(s) with high content of poly:unsaturated fatty acids |
| WO2009154309A1 (en) * | 2008-06-20 | 2009-12-23 | 有限会社梅田事務所 | Method for production of highly pure phospholipid, and highly pure sphingomyelin and plasmalogen-type glycerophospholipid produced by the method |
| US8524282B2 (en) | 2008-06-20 | 2013-09-03 | Umeda Jimusho Ltd. | Method for production of highly pure phospholipid, and highly pure sphingomyelin and plasmalogen-type glycerophospholipid produced by the method |
| JP5430566B2 (en) * | 2008-06-20 | 2014-03-05 | 有限会社梅田事務所 | Method for producing high purity phospholipid and high purity sphingomyelin and plasmalogen glycerophospholipid obtained by the method |
| JP2012170441A (en) * | 2011-02-24 | 2012-09-10 | Ueda Oils & Fats Mfg Co Ltd | Fish-derived phospholipid composition and method for producing the same |
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