JP6313627B2 - Powdered or solid composition containing acylated sterol glycoside and production method - Google Patents
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- 229930182558 Sterol Natural products 0.000 title claims description 18
- 235000003702 sterols Nutrition 0.000 title claims description 18
- -1 sterol glycoside Chemical class 0.000 title claims description 16
- 229930182470 glycoside Natural products 0.000 title claims description 15
- 238000004519 manufacturing process Methods 0.000 title claims description 9
- 239000008247 solid mixture Substances 0.000 title claims description 6
- 239000000203 mixture Substances 0.000 claims description 83
- 238000000605 extraction Methods 0.000 claims description 64
- 241000209094 Oryza Species 0.000 claims description 53
- 235000007164 Oryza sativa Nutrition 0.000 claims description 53
- 235000009566 rice Nutrition 0.000 claims description 53
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims description 45
- 239000002904 solvent Substances 0.000 claims description 43
- 229940067606 lecithin Drugs 0.000 claims description 42
- 235000010445 lecithin Nutrition 0.000 claims description 42
- 239000000787 lecithin Substances 0.000 claims description 42
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 36
- 239000002244 precipitate Substances 0.000 claims description 34
- 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 31
- 238000000194 supercritical-fluid extraction Methods 0.000 claims description 24
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 20
- 239000000725 suspension Substances 0.000 claims description 19
- 238000000034 method Methods 0.000 claims description 16
- 238000000926 separation method Methods 0.000 claims description 13
- 239000007788 liquid Substances 0.000 claims description 12
- 239000002994 raw material Substances 0.000 claims description 11
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 10
- 239000001569 carbon dioxide Substances 0.000 claims description 10
- 239000000284 extract Substances 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 7
- 150000003432 sterols Chemical class 0.000 claims description 3
- 239000004615 ingredient Substances 0.000 claims 2
- 150000003904 phospholipids Chemical class 0.000 description 21
- 238000004128 high performance liquid chromatography Methods 0.000 description 20
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 18
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 17
- 238000004458 analytical method Methods 0.000 description 10
- 238000004587 chromatography analysis Methods 0.000 description 10
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 10
- 239000000047 product Substances 0.000 description 10
- 238000010521 absorption reaction Methods 0.000 description 9
- 238000005119 centrifugation Methods 0.000 description 9
- 244000068988 Glycine max Species 0.000 description 7
- 235000010469 Glycine max Nutrition 0.000 description 7
- 238000000638 solvent extraction Methods 0.000 description 7
- 235000021329 brown rice Nutrition 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 5
- 238000003815 supercritical carbon dioxide extraction Methods 0.000 description 5
- 238000010828 elution Methods 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- 235000021588 free fatty acids Nutrition 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 239000012488 sample solution Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 125000005456 glyceride group Chemical group 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000002203 pretreatment Methods 0.000 description 2
- 238000010298 pulverizing process Methods 0.000 description 2
- 239000012856 weighed raw material Substances 0.000 description 2
- HNSDLXPSAYFUHK-UHFFFAOYSA-N 1,4-bis(2-ethylhexyl) sulfosuccinate Chemical compound CCCCC(CC)COC(=O)CC(S(O)(=O)=O)C(=O)OCC(CC)CCCC HNSDLXPSAYFUHK-UHFFFAOYSA-N 0.000 description 1
- 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 description 1
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonium chloride Substances [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 1
- 208000032131 Diabetic Neuropathies Diseases 0.000 description 1
- CWRILEGKIAOYKP-SSDOTTSWSA-M [(2r)-3-acetyloxy-2-hydroxypropyl] 2-aminoethyl phosphate Chemical compound CC(=O)OC[C@@H](O)COP([O-])(=O)OCCN CWRILEGKIAOYKP-SSDOTTSWSA-M 0.000 description 1
- 238000005377 adsorption chromatography Methods 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000003125 aqueous solvent Substances 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 239000002026 chloroform extract Substances 0.000 description 1
- 238000013375 chromatographic separation Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012847 fine chemical Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 150000008131 glucosides Chemical class 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 239000002035 hexane extract Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 235000011837 pasties Nutrition 0.000 description 1
- 150000008104 phosphatidylethanolamines Chemical class 0.000 description 1
- 239000003495 polar organic solvent Substances 0.000 description 1
- 238000002953 preparative HPLC Methods 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 150000003431 steroids Chemical group 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- 238000003260 vortexing Methods 0.000 description 1
- 230000002087 whitening effect Effects 0.000 description 1
Description
本発明は、アシル化ステロール配糖体を含有する粉末状又は固形状組成物及びその製造方法に関する。 The present invention relates to a powdery or solid composition containing an acylated sterol glycoside and a method for producing the same.
アシル化ステロール配糖体（以下「ASG」と記載する。）は、ステロイド骨格の3位に結合した糖がアシル化されている化合物である。このようなASGは大豆や玄米、発芽玄米などから得ることができる（特許文献１、特許文献２参照）。特許文献１の発芽玄米由来のASGは、糖尿病性神経障害の改善効果を有している。また特許文献２の大豆由来のASGは、DNA合成酵素の阻害作用を有している。 An acylated sterol glycoside (hereinafter referred to as “ASG”) is a compound in which a sugar bonded to the 3-position of the steroid skeleton is acylated. Such ASG can be obtained from soybean, brown rice, germinated brown rice, or the like (see Patent Document 1 and Patent Document 2). ASG derived from germinated brown rice in Patent Document 1 has an effect of improving diabetic neuropathy. Moreover, ASG derived from soybean of Patent Document 2 has an inhibitory action on DNA synthase.
これらのASGを得るためには、例えば特許文献１の発芽玄米由来のASGの場合は、発芽玄米を精白する際に得られる米ぬかから極性の低いクロロホルムや極性の高いメタノールなどの有機溶媒を用いてASG画分を抽出し（非特許文献１参照）、吸着クロマトグラフィー、さらには高速液体クロマトグラフィー（HPLC）を用いて調製している。
また、特許文献２の大豆由来のステロール配糖体の場合は、大豆（Glycine max L.）にn-ヘキサンを加えてn-ヘキサン抽出物を得た後、n-ヘキサンを留去し、クロロホルムを加え、クロロホルム抽出物を得て、さらにクロマト分離、HPLC操作を行って抽出する。
上記抽出方法を用いても、発芽玄米由来のASGの場合、乾燥物あたり 2.7%の含有量のものしか得られていない。またいずれもクロロホルムなどの溶媒を使用することが抽出効率をあげるための必須条件であり、クロロホルムを抽出溶媒として使用した場合、そのクロロホルムが抽出物に残存する。さらに溶媒抽出の場合、抽出溶媒中に米ぬかや大豆に含有される脂質がASGと同様の挙動を示すため、ASGと脂質の分離がはなはだ困難であり、溶媒抽出だけではASGの純度（含有率）を高めることが難しかった。
本発明者らは、このような取り扱い難いASGを含有する組成物を効率よく抽出する方法を検討し、超臨界抽出技術を応用した米糠からの二段階抽出技術を開発し既に特許出願を行っている（特願２０１２−１４６５７７号）。さらに発明者らは、超臨界抽出技術を応用した二段階抽出技術を用いて、ライスレシチンからASGを含有する粉末状の組成物を提供する技術においても既に特許出願を行っている（特願２０１３−１８０９０９）。しかし、上記の技術を用いることでライスレシチンに含まれる遊離脂肪酸およびグリセライド化合物を分離することは可能であるが、リン脂質を分離することは困難であり、得られるASG含有組成物のリン脂質含有率は57.8〜88.1%(w/w)である。上記の技術によって得られるASGは吸湿性が高く、その原因は両親媒性の特性を持つリン脂質であると考えられる。よってASGの高含有化および吸湿性改善のためにリン脂質分離の検討が必要であった。
In order to obtain these ASGs, for example, in the case of ASG derived from germinated brown rice of Patent Document 1, an organic solvent such as chloroform having a low polarity or methanol having a high polarity from rice bran obtained when whitening germinated brown rice is used. The ASG fraction is extracted (see Non-Patent Document 1) and prepared using adsorption chromatography and high performance liquid chromatography (HPLC).
In addition, in the case of the sterol glycoside derived from soybean of Patent Document 2, n-hexane was added to soybean (Glycine max L.) to obtain an n-hexane extract, and then n-hexane was distilled off, followed by chloroform. To obtain a chloroform extract, followed by further chromatographic separation and HPLC operation for extraction.
Even when the above extraction method is used, ASG derived from germinated brown rice has a content of only 2.7% per dry matter. In any case, the use of a solvent such as chloroform is an essential condition for increasing the extraction efficiency. When chloroform is used as an extraction solvent, the chloroform remains in the extract. Furthermore, in the case of solvent extraction, the lipid contained in rice bran and soybeans in the extraction solvent behaves in the same way as ASG, so it is difficult to separate ASG and lipid. ASG purity (content) It was difficult to increase.
The present inventors have studied a method for efficiently extracting a composition containing such difficult-to-handle ASG, developed a two-stage extraction technology from rice bran using supercritical extraction technology, and have already filed a patent application. (Japanese Patent Application No. 2012-146577). Furthermore, the inventors have already filed a patent application in a technique for providing a powdery composition containing ASG from rice lecithin using a two-stage extraction technique applying a supercritical extraction technique (Japanese Patent Application 2013). -180909). However, it is possible to separate free fatty acids and glyceride compounds contained in rice lecithin by using the above technique, but it is difficult to separate phospholipids, and the resulting ASG-containing composition contains phospholipids. The rate is 57.8-88.1% (w / w). ASG obtained by the above technique has high hygroscopicity, and the cause is considered to be phospholipids having amphiphilic properties. Therefore, it was necessary to study phospholipid separation in order to increase ASG content and improve hygroscopicity.
一般的なリン脂質を分離する方法として、エタノール（純度99〜90%(v/v)、その他の1〜10%(v/v)はほとんど水）を作用させて抽出する方法が知られている。また超臨界二酸化炭素抽出の際に、エントレーナーとしてエタノール（純度99.5%(v/v)、その他の0.5%(v/v)はほとんど水）を用いることでリン脂質を抽出する方法も知られている。しかし、上記のようなエタノールによる抽出方法では、ASGもエタノールと共に抽出されるため、得られる最終組成物は極めてASG含有率の低いものとなってしまう。よって、上記したような欠点を有する抽出方法に代えて、効率良く、リン脂質を分離でき、ASG含有率の高い組成物を提供しうる方法の開発が望まれているのが現状である。 As a general method for separating phospholipids, there is known a method of extraction with ethanol (purity 99 to 90% (v / v), other 1 to 10% (v / v) is almost water). Yes. Also known is a method of extracting phospholipids by using ethanol (purity 99.5% (v / v), other 0.5% (v / v) mostly water) as an entrainer during supercritical carbon dioxide extraction. ing. However, in the extraction method using ethanol as described above, since ASG is also extracted together with ethanol, the final composition obtained has a very low ASG content. Therefore, at present, it is desired to develop a method capable of efficiently separating phospholipids and providing a composition having a high ASG content, instead of the extraction method having the above-described drawbacks.
本発明は、高純度化された粉末状米由来ASGおよび吸湿性の低いASG含有組成物を提供することを課題とする。また本発明は、ライスレシチンオイルから遊離脂肪酸、グリセライド化合物およびリン脂質を効率的に分離する米由来のASG含有組成物の製造方法を提供することを課題とする。また本発明においてリン脂質とは、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、これらのリゾ化物（リゾホスファチジルコリン、リゾホスファチジルエタノールアミン）およびスフィンゴミエリンなどを構成成分とするものである。これらは類似した抽出挙動を示すため、本発明ではホスファチジルコリンを定量した。 An object of the present invention is to provide a highly purified powdery rice-derived ASG and an ASG-containing composition having low hygroscopicity. Another object of the present invention is to provide a method for producing a rice-derived ASG-containing composition that efficiently separates free fatty acids, glyceride compounds and phospholipids from rice lecithin oil. In the present invention, the phospholipid includes phosphatidylcholine, phosphatidylethanolamine, lysates thereof (lysophosphatidylcholine, lysophosphatidylethanolamine), sphingomyelin and the like as constituent components. Since these showed similar extraction behavior, phosphatidylcholine was quantified in the present invention.
本発明者らは、このような課題に即して研究を重ねた結果、より効率の良いASG含有組成物の製造技術を見出し、リン脂質含有量が低く、吸湿性の低いASG高含有組成物を得るに至った。
本発明は以下の構成である。
（１）ライスレシチン由来のアシル化ステロール配糖体含有組成物であって、ライスレシチン由来のアシル化ステロール配糖体を4%(w/w)以上含有し、ホスファチジルコリンが30%(w/w)以下であり、ライスレシチン由来以外の成分を含有しない粉末状又は固形状組成物。
（２）ライスレシチン由来のアシル化ステロール配糖体含有組成物であって、ライスレシチン由来のアシル化ステロール配糖体を8%(w/w)以上含有し、ホスファチジルコリンが20%(w/w)以下であり、ライスレシチン由来以外の成分を含有しない粉末状又は固形状組成物。
（３）40℃、75%RH条件下の恒温恒湿器内に一晩保管したとき重量増加率が5%(w/w)以下である（１）又は（２）に記載の組成物。
（４）米由来のライスレシチンを原料として粉末状又は固形状のライスレシチン由来のアシル化ステロール配糖体含有組成物を製造する方法であって、
ライスレシチンを、二酸化炭素を用いた超臨界抽出操作で処理し抽出残渣を得る工程、
得られた超臨界抽出残渣に抽出溶媒を加え、超臨界抽出物と抽出溶媒の懸濁液を調製する工程、
該懸濁液を固液分離する工程、
ライスレシチン由来のアシル化ステロール配糖体を含む不溶性画分（沈殿）を回収する工程、
を含むことを特徴とする（１）〜（３）のいずれかに記載の固形状又は粉末状のライスレシチン由来のアシル化ステロール配糖体含有組成物を製造する方法。
（５）抽出溶媒が含水エタノールである（４）記載の製造法。
As a result of repeated research in line with such problems, the present inventors have found a more efficient production technology for an ASG-containing composition, which has a low phospholipid content and a low hygroscopic content. I came to get.
The present invention has the following configuration.
(1) An acylated sterol glycoside-containing composition derived from rice lecithin, comprising 4% (w / w) or more of an acylated sterol glycoside derived from rice lecithin , and 30% (w / w) of phosphatidylcholine ) A powdery or solid composition which is the following and contains no components other than those derived from rice lecithin.
(2) An acylated sterol glycoside-containing composition derived from rice lecithin, comprising 8% (w / w) or more of acylated sterol glycoside derived from rice lecithin , and 20% (w / w) of phosphatidylcholine ) A powdery or solid composition which is the following and contains no components other than those derived from rice lecithin.
(3) The composition according to (1) or (2), wherein the weight increase rate is 5% (w / w) or less when stored overnight in a thermo-hygrostat under conditions of 40 ° C. and 75% RH.
(4) A method for producing a powdered or solid rice lecithin-derived acylated sterol glycoside- containing composition using rice-derived rice lecithin as a raw material,
A process of treating rice lecithin with a supercritical extraction operation using carbon dioxide to obtain an extraction residue;
Adding an extraction solvent to the resulting supercritical extraction residue to prepare a suspension of the supercritical extract and the extraction solvent;
Solid-liquid separation of the suspension;
Recovering an insoluble fraction (precipitate) containing an acylated sterol glycoside derived from rice lecithin ,
A method for producing an acylated sterol glycoside- containing composition derived from rice lecithin in solid or powder form according to any one of (1) to (3), which comprises:
(5) The production method according to (4), wherein the extraction solvent is hydrous ethanol.
本発明によって、粉末状又は固形状の米由来のASGであって、リン脂質含有量が低く吸湿性の低いASG含有組成物を提供が提供される。
また本発明のASG含有組成物はリン脂質が少なく、粉末にした場合の流動性が向上し、ASG含有組成物の打錠などの製剤化操作が容易となる利点がある。また吸湿性が低いため製剤化した際の安定性が高い。さらに組成物中のASG濃度が高くなるため、錠剤の小型化（小粒化）が可能となる。また、粉末化のための助剤を必要としないため、食品や医薬品原料として高濃度で利用可能なASG含有組成物となる。
さらに、本発明の製造方法は、超臨界抽出操作の回数が少ないため、簡便に製造できる特徴を有している。
The present invention provides an ASG-containing composition having a low phospholipid content and a low hygroscopicity, which is ASG derived from powdered or solid rice.
In addition, the ASG-containing composition of the present invention has an advantage that there are few phospholipids, the fluidity when powdered is improved, and formulation operations such as tableting of the ASG-containing composition are facilitated. Moreover, since it has low hygroscopicity, it has high stability when formulated. Furthermore, since the ASG concentration in the composition is increased, the tablet can be downsized (smaller). Moreover, since an auxiliary for pulverization is not required, an ASG-containing composition that can be used at a high concentration as a food or pharmaceutical raw material is obtained.
Furthermore, the production method of the present invention has a feature that it can be produced easily because the number of supercritical extraction operations is small.
以下本発明の実施形態を更に詳細に説明する。
粉末状又は固形状であって、米由来ASGを4% (w/w) 以上含有し、ホスファチジルコリンが30%(w/w)以下である組成物に係る発明である。
（１）原料
本発明の原料であるライスレシチンは、一般的に以下の方法により製造される。
米ぬかを加熱処理し、水分調整した後にヘキサンにより油分を抽出する。抽出した油分からヘキサンと不溶物を取り除き脱ガム処理として温水を加えてリン脂質を水和させる。その後、遠心分離器でガムと油を分離し、得られたガムがライスレシチンとなる。なお、ライスレシチンは約28.9〜43.3 % (w/w) のリン脂質を含有する。ライスレシチンはリン脂質を豊富に含有しており、レシチンの供給源としても利用されている。
Hereinafter, embodiments of the present invention will be described in more detail.
It is an invention relating to a composition which is in a powder or solid state and contains 4% (w / w) or more of rice-derived ASG and 30% (w / w) or less of phosphatidylcholine.
(1) Raw material Rice lecithin which is a raw material of the present invention is generally produced by the following method.
The rice bran is heat-treated and the water content is adjusted, and then the oil is extracted with hexane. Hexane and insoluble matter are removed from the extracted oil, and warm water is added as a degumming treatment to hydrate the phospholipid. Then, gum and oil are isolate | separated with a centrifuge, and the obtained gum turns into a rice lecithin. Rice lecithin contains about 28.9 to 43.3% (w / w) phospholipid. Rice lecithin is rich in phospholipids and is also used as a source of lecithin.
（２）超臨界抽出操作
ASGを抽出するために米油の精製工程で得られたライスレシチンを、超臨界二酸化炭素抽出装置の抽出槽に収納し、二酸化炭素の超臨界条件で抽出操作を行う。適切な抽出条件は、例えば、二酸化炭素流量 10〜100g/分、抽出時間1〜5時間、抽出圧力10〜50MPaおよび抽出温度32〜70℃である。より好ましくは二酸化炭素流量65g/分、抽出時間4時間、抽出圧力25MPaおよび抽出温度40℃であり、さらに好ましくは二酸化炭素流量65g/分、抽出時間4時間、抽出圧力50MPaおよび抽出温度70℃である。これにより、遊離脂肪酸およびグリセライド化合物が分離された超臨界二酸化炭素抽出残渣が得られる。
(2) Supercritical extraction operation
Rice lecithin obtained in the rice oil refining process to extract ASG is stored in an extraction tank of a supercritical carbon dioxide extraction device, and extraction operation is performed under supercritical conditions of carbon dioxide. Suitable extraction conditions are, for example, a carbon dioxide flow rate of 10-100 g / min, an extraction time of 1-5 hours, an extraction pressure of 10-50 MPa and an extraction temperature of 32-70 ° C. More preferably, the carbon dioxide flow rate is 65 g / min, the extraction time is 4 hours, the extraction pressure is 25 MPa, and the extraction temperature is 40 ° C., and the carbon dioxide flow rate is 65 g / min, the extraction time is 4 hours, the extraction pressure is 50 MPa, and the extraction temperature is 70 ° C. is there. Thereby, the supercritical carbon dioxide extraction residue from which the free fatty acid and the glyceride compound are separated is obtained.
（３）溶媒抽出
第一段階の超臨界抽出が終了した抽出槽の残渣は、極性を有する有機溶媒（たとえばアセトン、メタノール、エタノールおよびイソプロパノール）と水の混合溶液で溶媒抽出を行う。有機溶媒は特にエタノールが好ましい。
適切な抽出条件として例えばエタノール50〜80%(v/v)、水50〜20%(v/v)の混合溶媒を調製し、この溶媒による振盪抽出または還流抽出を行う。特に好ましくはエタノール70%(v/v)、水30%(v/v)の比率の溶媒とする。抽出操作は、超臨界二酸化炭素抽出残渣と溶媒をよく混合し、懸濁状態にする。懸濁させる時間は長い方が好ましい。例えば、一時間、より好ましくは一晩である。加える溶媒量が多い方がリン脂質の抽出効率が高まり、分離を良くすることができる。また、超臨界二酸化炭素抽出残渣は溶媒抽出の前に、予め粉砕処理を行うことが好ましい。抽出処理を複数回繰り返すことでより多くのリン脂質を抽出除去することができる。
(3) Solvent extraction The residue in the extraction tank after the first stage of supercritical extraction is subjected to solvent extraction with a mixed solution of polar organic solvent (for example, acetone, methanol, ethanol and isopropanol) and water. The organic solvent is particularly preferably ethanol.
As a suitable extraction condition, for example, a mixed solvent of ethanol 50 to 80% (v / v) and water 50 to 20% (v / v) is prepared, and shake extraction or reflux extraction with this solvent is performed. Particularly preferred is a solvent having a ratio of ethanol 70% (v / v) and water 30% (v / v). In the extraction operation, the supercritical carbon dioxide extraction residue and the solvent are mixed well to make a suspension. A longer suspension time is preferable. For example, one hour, more preferably overnight. When the amount of the solvent added is large, the extraction efficiency of the phospholipid is increased and the separation can be improved. The supercritical carbon dioxide extraction residue is preferably pulverized in advance before solvent extraction. More phospholipids can be extracted and removed by repeating the extraction process a plurality of times.
（４）固液分離
所望する時間経過後、懸濁液を遠心分離、濾過等により固液分離する。この際、リン脂質を可溶性画分にできるだけ移行させる一方、米由来ASGの大部分は可溶性画分に溶出させず、沈殿（不溶性画分）として回収することが重要である。そのためには沈殿をできる限り再懸濁させないように注意が必要である。
得られた不溶性画分は、必要により凍結乾燥などの減圧乾燥処理を施し、残存する溶媒を除去してもよい。
かくして得られる組成物は、粉末状を呈しており、リン脂質含有率が低く（特にホスファチジルコリン含有率は30%(w/w)以下である）、ASGを4〜8%(w/w)以上含有している。この組成物をカラムクロマトグラフィーや分取型高速液体クロマトグラフィー装置（HPLC）にかけてさらにASG含有率を高めた組成物を得るための原料とすることもできる。
(4) Solid-liquid separation After the desired time has elapsed, the suspension is subjected to solid-liquid separation by centrifugation, filtration, or the like. At this time, it is important that the phospholipid is transferred to the soluble fraction as much as possible, but most of the rice-derived ASG is not eluted in the soluble fraction but recovered as a precipitate (insoluble fraction). For this purpose, care must be taken not to resuspend the precipitate as much as possible.
The obtained insoluble fraction may be subjected to vacuum drying treatment such as lyophilization, if necessary, to remove the remaining solvent.
The composition thus obtained is in the form of a powder, has a low phospholipid content (particularly the phosphatidylcholine content is 30% (w / w) or less), and has an ASG content of 4-8% (w / w) or more. Contains. This composition can also be used as a raw material for obtaining a composition having a further increased ASG content by subjecting it to column chromatography or preparative high-performance liquid chromatography (HPLC).
（５）原料及び組成物中の分析・ASGおよびホスファチジルコリンの確認方法
原料及び得られたASG含有組成物中のASG含有量は以下に示す方法で分析を行い確認する。
１）ライスレシチンの前処理方法
i） 50mLの蓋付ガラスチューブにライスレシチンを1.0g正秤する。
ii) 次いで、クロロホルム：メタノール＝2：1(v/v)の混合溶液（以下クロメタ混液）を10mL加え、ボルテックスにて約90秒間、よく懸濁・分散させた後、遠心分離（室温、1,500×g、10分）し、上澄液を回収する。
iii)ii)の操作をさらに2回繰り返し、フィルター濾過（目開き：0.45μm）をし、その濾液を分析用試料溶液とする。（得られた試料溶液は成分濃度に応じて適宜希釈を行う。希釈液はクロメタ混液を用いる。）
２）ASG含有組成物の前処理方法
i) 得られたASG含有組成物を適量秤取し、クロメタ混液を加え、50mLメスフラスコにてメスアップする。
ii) i)で得られた溶液は成分濃度に応じて適宜希釈し、分析用試料溶液とする。
(5) Analysis in raw material and composition-Method for confirming ASG and phosphatidylcholine The ASG content in the raw material and the obtained ASG-containing composition is analyzed and confirmed by the following method.
1) Pretreatment method of rice lecithin
i) Weigh 1.0 g of rice lecithin into a 50 mL glass tube with a lid.
ii) Next, 10 mL of a mixed solution of chloroform: methanol = 2: 1 (v / v) (hereinafter referred to as “chromometa mixture”) was added and suspended and dispersed well by vortexing for about 90 seconds, followed by centrifugation (room temperature, 1,500 Xg, 10 minutes) and collect the supernatant.
iii) Repeat step ii) two more times, filter through a filter (opening: 0.45 μm), and use the filtrate as a sample solution for analysis. (The obtained sample solution is appropriately diluted according to the component concentration. The dilution liquid is a chromite mixture.)
2) Pretreatment method for ASG-containing composition
i) Weigh an appropriate amount of the obtained ASG-containing composition, add the chromometa mixture, and measure up in a 50 mL volumetric flask.
ii) Dilute the solution obtained in i) as appropriate according to the component concentration to make the sample solution for analysis.
３）ASG分析方法
i)HPLCによる分析
HPLC分析条件は次のとおり
分析装置：HPLC
移動相Ａ：メタノール：水 = 95：5 (v/v)
移動相Ｂ：クロロホルム = 100 (v/v)
ポンプ：Model 582 solvent delivery system
カラム：LiChrospher Si60 (5μm) HPLC-Cartridge (LiChroCART125-4:MERCK)
検出器：荷電化粒子検出器（コロナダイオネクス社）
Injection Volume：20μL
カラムオーブン：40℃（FLO社 model 502）
分析時間：40分
脱泡装置：Uniflows Degasys Ultimate DV3003
流速：1mL/分
ii)グラジエント条件
以下に示すとおり
(1)0-15分
移動相A：1%(v/v) → 25%(v/v)、移動相B：99%(v/v) → 75%(v/v)
(2)15-20分
移動相A：25%(v/v) → 90%(v/v)、移動相B：75%(v/v) → 10%(v/v)
(3)20-25分
移動相A：90%(v/v)、移動相B：10%(v/v)
(4)25-30分
移動相A：90%(v/v) → 1%(v/v)、移動相B：10%(v/v) → 99%(v/v)
(5)30-40分
移動相A：1%(v/v)、移動相B：99%(v/v)
iii)標準品
ASGの標準品として、Esterified Steryl Glucoside（Larodan Fine Chemicals AB製、フナコシ社販売）を用いる。標準品2mgはアンプル開封後、窒素乾固させクロメタ混液で200μg/mLの濃度となるように溶解し、段階希釈して100、50、25、12.5及び 6.25μg/mLの濃度系列とし、検量線を作成する。
3) ASG analysis method
i) Analysis by HPLC
HPLC analysis conditions are as follows Analyzer: HPLC
Mobile phase A: methanol: water = 95: 5 (v / v)
Mobile phase B: chloroform = 100 (v / v)
Pump: Model 582 solvent delivery system
Column: LiChrospher Si60 (5μm) HPLC-Cartridge (LiChroCART125-4: MERCK)
Detector: Charged particle detector (Corona Dionex)
Injection Volume: 20μL
Column oven: 40 ° C (FLO model 502)
Analysis time: 40 minutes Deaerator: Uniflows Degasys Ultimate DV3003
Flow rate: 1mL / min
ii) Gradient conditions As shown below
(1) 0-15 minutes Mobile phase A: 1% (v / v) → 25% (v / v), Mobile phase B: 99% (v / v) → 75% (v / v)
(2) 15-20 minutes Mobile phase A: 25% (v / v) → 90% (v / v), Mobile phase B: 75% (v / v) → 10% (v / v)
(3) 20-25 minutes Mobile phase A: 90% (v / v), Mobile phase B: 10% (v / v)
(4) 25-30 minutes Mobile phase A: 90% (v / v) → 1% (v / v), Mobile phase B: 10% (v / v) → 99% (v / v)
(5) 30-40 minutes Mobile phase A: 1% (v / v), Mobile phase B: 99% (v / v)
iii) Standard product
Esterified Steryl Glucoside (manufactured by Larodan Fine Chemicals AB, sold by Funakoshi) is used as a standard product of ASG. After opening the ampoule, 2 mg of the standard product is nitrogen-dried, dissolved in a chromometa mixture to a concentration of 200 μg / mL, serially diluted to a concentration series of 100, 50, 25, 12.5 and 6.25 μg / mL. Create
４）ホスファチジルコリン分析方法
i)HPLCによる分析
HPLC分析条件は次のとおり
分析装置：HPLC
移動相Ａ：クロロホルム：メタノール：25%アンモニア水溶液= 160：39：1 (v/v)
移動相Ｂ：クロロホルム：メタノール：水：25%アンモニア水溶液= 120：68：11：1 (v/v)カラム：LiChrospher Si60 (5μm) HPLC-Cartridge (LiChroCART125-4:MERCK)
Injection Volume：20μL
カラムオーブン：25℃（FLO社model 502）
分析時間：30分
流速 ：1mL/分
脱泡装置：Uniflows Degasys Ultimate DV3003
ii)グラジエント条件
以下に示すとおり
(1)0-1分
移動相A：80%(v/v)、移動相B：20%(v/v)
(2)1-8分
移動相A：80%(v/v) → 1%(v/v)、移動相B：20%(v/v) → 99%(v/v)
(3)8-20分
移動相A：1%(v/v)、移動相B：99%(v/v)
(4)20-30分
移動相A：1%(v/v) → 80%(v/v)、移動相B：99%(v/v) → 20%(v/v)
iii) 標準品
ホスファチジルコリンの標準品として、L-α-ホスファチジルコリン、 大豆由来、水素添加（和光純薬工業社販売）を用いる。標準品を秤量し200μg/mLの濃度となるようにクロメタ混液に溶解させる、段階希釈して100、50、25、12.5及び 6.25μg/mLの濃度系列とし、検量線を作成する。
4) Phosphatidylcholine analysis method
i) Analysis by HPLC
HPLC analysis conditions are as follows Analyzer: HPLC
Mobile phase A: chloroform: methanol: 25% aqueous ammonia solution = 160: 39: 1 (v / v)
Mobile phase B: Chloroform: Methanol: Water: 25% ammonia aqueous solution = 120: 68: 11: 1 (v / v) Column: LiChrospher Si60 (5 μm) HPLC-Cartridge (LiChroCART125-4: MERCK)
Injection Volume: 20μL
Column oven: 25 ° C (FLO model 502)
Analysis time: 30 minutes Flow rate: 1 mL / min Defoaming device: Uniflows Degasys Ultimate DV3003
ii) Gradient conditions As shown below
(1) 0-1 minutes Mobile phase A: 80% (v / v), Mobile phase B: 20% (v / v)
(2) 1-8 minutes Mobile phase A: 80% (v / v) → 1% (v / v), Mobile phase B: 20% (v / v) → 99% (v / v)
(3) 8-20 minutes Mobile phase A: 1% (v / v), Mobile phase B: 99% (v / v)
(4) 20-30 minutes Mobile phase A: 1% (v / v) → 80% (v / v), Mobile phase B: 99% (v / v) → 20% (v / v)
iii) Standard product As a standard product of phosphatidylcholine, L-α-phosphatidylcholine, derived from soybean, hydrogenated (sold by Wako Pure Chemical Industries, Ltd.) is used. Weigh a standard sample and dissolve it in a chromometa mixture to a concentration of 200 μg / mL, serially dilute into 100, 50, 25, 12.5 and 6.25 μg / mL concentration series, and create a calibration curve.
以下に実施例、比較例を示し、本発明をさらに詳細に説明する。
（実施例１）
ライスレシチンを約60g採取し、これを超臨界抽出装置（三菱化工機社製）の耐圧容器（抽出槽，0.5L容器）に移した。次いで抽出溶媒として二酸化炭素を用いて、昇温・昇圧操作を行い、70℃、50MPaで4時間（CO2流速：65g/分）、超臨界二酸化炭素を抽出槽に通して抽出操作を行った。
抽出液は回収槽に移し、秤量した。本条件での抽出操作により、秤量した原料の約60%が抽出され、約40%が残渣として抽出槽から回収できた。この組成物のASG含有率は2.84%、ホスファチジルコリン含有率は22.86%(w/w)であった。
この組成物を40℃、75%RHの恒温恒湿器内で一晩保管した際の、吸湿による重量増加率（以下、「吸湿率」と記載する。）は11.10%(w/w)であった。
またこの組成物は固形状であり、粉砕時の流動性はあまり良くなかった。
図３に原料としたライスレシチンのASG含量を分析するために実施したHPLCクロマトグラフィーのチャートを示す。図４に実施例１で得たASG含有組成物のHPLCクロマトパターンを示す。さらに、図６にライスレシチンのホスファチジルコリン含有量を分析するために実施したHPLCクロマトグラフィーのチャートを、図７に実施例１で得た組成物のホスファチジルコリン含有量を分析するために実施したHPLCクロマトグラフィーのチャートを示す。
ライスレシチンに含有されているASGおよびリン脂質以外の成分が低下していることがわかる。
Hereinafter, the present invention will be described in more detail with reference to Examples and Comparative Examples.
Example 1
About 60 g of rice lecithin was collected and transferred to a pressure-resistant container (extraction tank, 0.5 L container) of a supercritical extraction device (manufactured by Mitsubishi Chemical Corporation). Next, using carbon dioxide as an extraction solvent, the temperature was increased and the pressure was increased, and the extraction operation was performed by passing supercritical carbon dioxide through the extraction tank at 70 ° C. and 50 MPa for 4 hours (CO 2 flow rate: 65 g / min). .
The extract was transferred to a collection tank and weighed. By the extraction operation under these conditions, about 60% of the weighed raw material was extracted, and about 40% was recovered as a residue from the extraction tank. This composition had an ASG content of 2.84% and a phosphatidylcholine content of 22.86% (w / w).
When this composition was stored overnight in a constant temperature and humidity chamber at 40 ° C. and 75% RH, the weight increase rate due to moisture absorption (hereinafter referred to as “moisture absorption rate”) was 11.10% (w / w). there were.
Moreover, this composition was solid and the fluidity | liquidity at the time of a grinding | pulverization was not so good.
FIG. 3 shows a chart of HPLC chromatography performed to analyze the ASG content of rice lecithin as a raw material. FIG. 4 shows an HPLC chromatogram pattern of the ASG-containing composition obtained in Example 1. Further, FIG. 6 shows a chart of HPLC chromatography performed for analyzing the phosphatidylcholine content of rice lecithin, and FIG. 7 shows an HPLC chromatography performed for analyzing the phosphatidylcholine content of the composition obtained in Example 1. The chart is shown.
It can be seen that components other than ASG and phospholipid contained in rice lecithin are reduced.
（実施例２）
実施例１で得られた超臨界抽出残渣を粉砕し、この粉砕物100mgに対して、エタノール：水＝7：3(v/v)の抽出溶媒を50mL加えて、懸濁液とし、常温で1時間振盪し、溶媒抽出した。振盪後、固液分離操作を行った。遠心分離を3,000×gで5分間行い、懸濁液を可溶性画分と沈殿に分離させ、沈殿を回収した。この沈殿を減圧下で溶媒を除去し、ASG含有組成物とした。
回収したASG含有組成物は、上記の超臨界抽出残渣の総重量の約75%が溶媒側に移行しており、回収した沈殿物は抽出残渣重量の約25%であった。
この沈殿（ASG含有組成物）は、ASGは8.83%(w/w)、ホスファチジルコリン含有率は19.76%(w/w)であった。
またこの組成物を40℃、75%RH条件下の恒温恒湿器内で一晩保管した際の、吸湿率は1.57%(w/w)であった。さらに得られた組成物は粉末状であり流動性に優れていた。
(Example 2)
The supercritical extraction residue obtained in Example 1 was pulverized, and 50 mL of ethanol: water = 7: 3 (v / v) extraction solvent was added to 100 mg of this pulverized product to form a suspension. Shake for 1 hour and extract with solvent. After shaking, solid-liquid separation operation was performed. Centrifugation was performed at 3,000 × g for 5 minutes to separate the suspension into a soluble fraction and a precipitate, and the precipitate was collected. From this precipitate, the solvent was removed under reduced pressure to obtain an ASG-containing composition.
In the recovered ASG-containing composition, about 75% of the total weight of the supercritical extraction residue was transferred to the solvent side, and the recovered precipitate was about 25% of the extraction residue weight.
This precipitate (ASG-containing composition) had an ASG content of 8.83% (w / w) and a phosphatidylcholine content of 19.76% (w / w).
When this composition was stored overnight in a thermo-hygrostat under the conditions of 40 ° C. and 75% RH, the moisture absorption rate was 1.57% (w / w). Furthermore, the obtained composition was powdery and excellent in fluidity.
（実施例３）
実施例１で得られた超臨界抽出残渣を粉砕し、この粉砕物100mgに対して、エタノール：水＝7：3(v/v)の抽出溶媒を50mL加えて、懸濁液とし、常温で1時間振盪し、溶媒抽出した。振盪後、固液分離操作を行った。遠心分離を3,000×gで5分間行い、懸濁液を可溶性画分と沈殿に分離させ、沈殿を回収した。
得られた沈殿に再度エタノール：水＝7：3(v/v)の抽出溶媒を50mL加えて溶媒抽出および固液分離操作を4回繰り返した。回収した沈殿を減圧下で溶媒を除去し、ASG含有組成物とした。このASG含有組成物は、超臨界抽出残渣の総重量の約80%溶媒側に移行し、回収した沈殿物は抽出残渣重量の約20%であった。
このASG含有組成物は、ホスファチジルコリンをほとんど含んでいなかった(0.63%)。またASG含有率は9.68%(w/w)であった。
この組成物を40℃、75%RH条件下の恒温恒湿器内で一晩保管した際の、吸湿率は1.21%(w/w)であった。さらに得られた組成物は粉末状であり流動性に優れていた。
図５に実施例３の組成物のASG含有量を測定するために行ったHPLCクロマトグラフィーのチャートを、図８に同じくホスファチジルコリン含有量を測定するために行ったHPLC分析のクロマトグラフィーのチャートを示す。ASGに対してリン脂質ピークが低下し、ホスファチジルコリンが検出されなくなったことがわかる。
(Example 3)
The supercritical extraction residue obtained in Example 1 was pulverized, and 50 mL of ethanol: water = 7: 3 (v / v) extraction solvent was added to 100 mg of this pulverized product to form a suspension. Shake for 1 hour and extract with solvent. After shaking, solid-liquid separation operation was performed. Centrifugation was performed at 3,000 × g for 5 minutes to separate the suspension into a soluble fraction and a precipitate, and the precipitate was collected.
50 mL of an extraction solvent of ethanol: water = 7: 3 (v / v) was again added to the obtained precipitate, and solvent extraction and solid-liquid separation were repeated 4 times. The collected precipitate was subjected to removal of the solvent under reduced pressure to obtain an ASG-containing composition. The ASG-containing composition moved to the solvent side of about 80% of the total weight of the supercritical extraction residue, and the recovered precipitate was about 20% of the weight of the extraction residue.
This ASG-containing composition contained little phosphatidylcholine (0.63%). The ASG content was 9.68% (w / w).
When this composition was stored overnight in a thermo-hygrostat under conditions of 40 ° C. and 75% RH, the moisture absorption rate was 1.21% (w / w). Furthermore, the obtained composition was powdery and excellent in fluidity.
FIG. 5 shows a chart of HPLC chromatography performed for measuring the ASG content of the composition of Example 3, and FIG. 8 also shows a chromatographic chart of HPLC analysis performed for measuring the phosphatidylcholine content. . It can be seen that the phospholipid peak decreased with respect to ASG, and phosphatidylcholine was no longer detected.
（実施例４）
実施例１で得られた超臨界抽出残渣を粉砕し、この粉砕物100mgに対して、エタノール：水＝8：2(v/v)の抽出溶媒を50mL加えて、懸濁液とし、常温で1時間振盪し、溶媒抽出した。振盪後、固液分離操作を行った。遠心分離を3,000×gで5分間行い、懸濁液を可溶性画分と沈殿に分離させ、沈殿を回収し、減圧下で溶媒を除去し、ASG含有組成物とした。
このASG含有組成物は、超臨界抽出残渣の総重量の約75%が溶媒側に移行し、回収した沈殿物は抽出残渣重量の約25%であった。
このASG含有組成物のASG含有率は5.71%(w/w)であった。またホスファチジルコリン含有率は8.12%(w/w)であった。
さらにまた、この組成物を40℃、75%RHの恒温恒湿器内で一晩保管した際の、吸湿率は2.56%(w/w)であった。さらに得られた組成物は粉末状であり流動性に優れていた。
Example 4
The supercritical extraction residue obtained in Example 1 was pulverized, and 50 mL of an extraction solvent of ethanol: water = 8: 2 (v / v) was added to 100 mg of this pulverized product to form a suspension. Shake for 1 hour and extract with solvent. After shaking, solid-liquid separation operation was performed. Centrifugation was performed at 3,000 × g for 5 minutes to separate the suspension into a soluble fraction and a precipitate. The precipitate was collected, and the solvent was removed under reduced pressure to obtain an ASG-containing composition.
In this ASG-containing composition, about 75% of the total weight of the supercritical extraction residue was transferred to the solvent side, and the recovered precipitate was about 25% of the extraction residue weight.
The ASG content of this ASG-containing composition was 5.71% (w / w). The phosphatidylcholine content was 8.12% (w / w).
Furthermore, when this composition was stored overnight in a constant temperature and humidity chamber at 40 ° C. and 75% RH, the moisture absorption rate was 2.56% (w / w). Furthermore, the obtained composition was powdery and excellent in fluidity.
（実施例５）
実施例１で得られた超臨界抽出残渣を粉砕し、この粉砕物100mgに対して、エタノール：水＝6：4(v/v)の抽出溶媒を50mL加えて、懸濁液とし、常温で1時間振盪し、溶媒抽出した。振盪後、固液分離操作を行った。遠心分離を3,000×gで5分間行い、懸濁液を可溶性画分と沈殿に分離させ、沈殿を回収した。
回収した沈殿を減圧下で溶媒を除去し、ASG含有組成物とした。このASG含有組成物は、超臨界抽出残渣の総重量の約60%が溶媒側に移行し、回収した沈殿物は抽出残渣重量の約40%であった。
このASG含有組成物は、ASG含有率は5.24%(w/w)であった。またホスファチジルコリン含有率は21.14%(w/w)であった。
さらにまた、この組成物を40℃、75%RH条件下の恒温恒湿器内で一晩保管した際の、吸湿率は4.85%(w/w)であった。さらに得られた組成物は粉末状であり流動性に優れていた。
(Example 5)
The supercritical extraction residue obtained in Example 1 was pulverized, and 50 mL of an extraction solvent of ethanol: water = 6: 4 (v / v) was added to 100 mg of this pulverized product to form a suspension. Shake for 1 hour and extract with solvent. After shaking, solid-liquid separation operation was performed. Centrifugation was performed at 3,000 × g for 5 minutes to separate the suspension into a soluble fraction and a precipitate, and the precipitate was collected.
The collected precipitate was subjected to removal of the solvent under reduced pressure to obtain an ASG-containing composition. In this ASG-containing composition, about 60% of the total weight of the supercritical extraction residue was transferred to the solvent side, and the recovered precipitate was about 40% of the extraction residue weight.
This ASG-containing composition had an ASG content of 5.24% (w / w). The phosphatidylcholine content was 21.14% (w / w).
Furthermore, when this composition was stored overnight in a thermo-hygrostat under conditions of 40 ° C. and 75% RH, the moisture absorption rate was 4.85% (w / w). Furthermore, the obtained composition was powdery and excellent in fluidity.
（実施例６）
ライスレシチンを約60g採取し、これを超臨界抽出装置（三菱化工機社製）の耐圧容器（抽出槽，0.5L容器）に移した。次いで抽出溶媒として二酸化炭素を用いて、昇温・昇圧操作を行い、40℃、25MPaで4時間（CO2流速：65g/分）、超臨界二酸化炭素を抽出槽に通して抽出操作を行った。
抽出液は回収槽に移し、秤量した。本条件での抽出操作により、秤量した原料の約60%が抽出され、約40%が残渣として抽出槽から回収できた。こうして得られたASG含有組成物は、ホスファチジルコリンの含有率24.86%(w/w)、ASG含有率は2.77%(w/w)であった。
この抽出残渣を粉砕し、抽出残渣重量約100mgに対して、エタノール：水＝7：3(v/v)の抽出溶媒を50mL加えて、懸濁液とし、常温で1時間振盪し、溶媒抽出した。
振盪後、遠心分離を3,000×gにて5分間行い、可溶性画分と沈殿に分離させて、沈殿を回収した。この沈殿を減圧下で溶媒除去し、ASG組成物とした。このASG含有組成物は超臨界抽出残渣の総重量の約60%が溶媒側に移行し、抽出残渣重量の約40%が沈殿物として回収された。
この組成物のASG含有率は、5.44%(w/w)、ホスファチジルコリン含有率は10.25%(w/w)であった。
また、この組成物を40℃、75%RH条件下の恒温恒湿器内で一晩保管した際の、吸湿率は3.98%(w/w)であった。さらに得られた組成物は粉末状であり流動性に優れていた。
(Example 6)
About 60 g of rice lecithin was collected and transferred to a pressure-resistant container (extraction tank, 0.5 L container) of a supercritical extraction device (manufactured by Mitsubishi Chemical Corporation). Next, using carbon dioxide as the extraction solvent, the temperature was increased and the pressure was increased. The extraction operation was performed by passing supercritical carbon dioxide through the extraction tank at 40 ° C and 25 MPa for 4 hours (CO 2 flow rate: 65 g / min). .
The extract was transferred to a collection tank and weighed. By the extraction operation under these conditions, about 60% of the weighed raw material was extracted, and about 40% was recovered as a residue from the extraction tank. The ASG-containing composition thus obtained had a phosphatidylcholine content of 24.86% (w / w) and an ASG content of 2.77% (w / w).
This extraction residue is pulverized, and 50 mL of ethanol: water = 7: 3 (v / v) is added to about 100 mg of the extraction residue to make a suspension. The suspension is shaken at room temperature for 1 hour, and extracted with solvent. did.
After shaking, centrifugation was performed at 3,000 × g for 5 minutes to separate the precipitate into a soluble fraction and a precipitate, and the precipitate was collected. This precipitate was subjected to solvent removal under reduced pressure to obtain an ASG composition. In this ASG-containing composition, about 60% of the total weight of the supercritical extraction residue was transferred to the solvent side, and about 40% of the extraction residue weight was recovered as a precipitate.
The composition had an ASG content of 5.44% (w / w) and a phosphatidylcholine content of 10.25% (w / w).
In addition, when this composition was stored overnight in a thermo-hygrostat under conditions of 40 ° C. and 75% RH, the moisture absorption rate was 3.98% (w / w). Furthermore, the obtained composition was powdery and excellent in fluidity.
（比較例１）溶媒抽出のみによるASG組成物の調製例
ライスレシチンを約100mg採取し、エタノール：水＝7：3(v/v)の抽出溶媒を50mL加えて、懸濁液とし、常温で一時間振盪し、溶媒抽出した。振盪後の固液分離操作として、遠心分離を3,000×gにて5分間行い、可溶性画分と不溶性画分（沈殿）に分離させ沈殿を得た。この沈殿から減圧下で溶媒を除去した。この操作によって、秤量したライスレシチンに対して約50%が除去され、残りの約50%を沈殿物として得た。こうして得られたASG含有組成物は、ASG含有率は0.36%(w/w)、ホスファチジルコリン含有率は0.63%(w/w)、であった。
また、この組成物を40℃、75%RHの恒温恒湿器内で一晩保管した際の、吸湿率は1.86%(w/w)であった。さらに得られた組成物はペースト状で流動性が悪かった。
(Comparative example 1) Preparation example of ASG composition only by solvent extraction About 100 mg of rice lecithin was collected, and 50 mL of ethanol: water = 7: 3 (v / v) extraction solvent was added to form a suspension at room temperature. Shake for 1 hour and extract with solvent. As a solid-liquid separation operation after shaking, centrifugation was performed at 3,000 × g for 5 minutes to separate a soluble fraction and an insoluble fraction (precipitate) to obtain a precipitate. The solvent was removed from the precipitate under reduced pressure. By this operation, about 50% of the weighed rice lecithin was removed, and the remaining about 50% was obtained as a precipitate. The ASG-containing composition thus obtained had an ASG content of 0.36% (w / w) and a phosphatidylcholine content of 0.63% (w / w).
In addition, when this composition was stored overnight in a constant temperature and humidity chamber at 40 ° C. and 75% RH, the moisture absorption rate was 1.86% (w / w). Further, the obtained composition was pasty and poor in fluidity.
（比較例２）超臨界抽出後非含水溶媒で抽出した調製例
実施例１で得られた超臨界抽出残渣を粉砕し、この粉砕物100mgに対して、無水エタノール50mLを加えて、懸濁液とし、常温で1時間振盪し、溶媒抽出した。振盪後、固液分離操作を行った。遠心分離を3,000×gで5分間行い、懸濁液を可溶性画分と沈殿に分離させ、沈殿を回収した。この沈殿を減圧下で溶媒を除去し、ASG含有組成物とした。
回収したASG含有組成物は、上記の超臨界抽出残渣の総重量の約70%が溶媒側に移行しており、回収した沈殿は抽出残渣重量の約30%であった。
この組成物は、ASG含有率0.56%(w/w)、ホスファチジルコリン含有率は15.20%(w/w)であった。また、この組成物を40℃、75%RHの恒温恒湿器内で一晩保管した際の、吸湿率は2.27%(w/w)であった。さらに得られたASG含有組成物は粉末状であり流動性に優れていた。
以上の実施例、比較例の分析結果、評価結果を下記の表1にまとめて示す。
(Comparative example 2) Preparation example extracted with non-aqueous solvent after supercritical extraction The supercritical extraction residue obtained in Example 1 was pulverized, and 50 mL of absolute ethanol was added to 100 mg of this pulverized product, The mixture was shaken at room temperature for 1 hour and extracted with a solvent. After shaking, solid-liquid separation operation was performed. Centrifugation was performed at 3,000 × g for 5 minutes to separate the suspension into a soluble fraction and a precipitate, and the precipitate was collected. From this precipitate, the solvent was removed under reduced pressure to obtain an ASG-containing composition.
In the recovered ASG-containing composition, about 70% of the total weight of the supercritical extraction residue was transferred to the solvent side, and the recovered precipitate was about 30% of the extraction residue weight.
This composition had an ASG content of 0.56% (w / w) and a phosphatidylcholine content of 15.20% (w / w). In addition, when this composition was stored overnight in a constant temperature and humidity chamber at 40 ° C. and 75% RH, the moisture absorption rate was 2.27% (w / w). Further, the obtained ASG-containing composition was powdery and excellent in fluidity.
The analysis results and evaluation results of the above Examples and Comparative Examples are summarized in Table 1 below.
比較例１、２から本発明の組成物を得るためには、超臨界抽出と含水有機溶媒による溶媒抽出が必須であることが判明した。 In order to obtain the composition of the present invention from Comparative Examples 1 and 2, it was found that supercritical extraction and solvent extraction with a water-containing organic solvent are essential.
Claims (5)
ライスレシチンを、二酸化炭素を用いた超臨界抽出操作で処理し抽出残渣を得る工程、
得られた超臨界抽出残渣に抽出溶媒を加え、超臨界抽出物と抽出溶媒の懸濁液を調製する工程、
該懸濁液を固液分離する工程、
ライスレシチン由来のアシル化ステロール配糖体を含む不溶性画分（沈殿）を回収する工程、
を含むことを特徴とする請求項１〜３のいずれかに記載の固形状又は粉末状のライスレシチン由来のアシル化ステロール配糖体含有組成物を製造する方法。 A method of producing a powdered or solid rice lecithin-derived acylated sterol glycoside- containing composition using rice-derived rice lecithin as a raw material,
A process of treating rice lecithin with a supercritical extraction operation using carbon dioxide to obtain an extraction residue;
Adding an extraction solvent to the resulting supercritical extraction residue to prepare a suspension of the supercritical extract and the extraction solvent;
Solid-liquid separation of the suspension;
Recovering an insoluble fraction (precipitate) containing an acylated sterol glycoside derived from rice lecithin ,
The method of manufacturing the acylated sterol glycoside containing composition derived from the solid or powdery rice lecithin in any one of Claims 1-3 characterized by the above-mentioned.
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