JPS625917A - Production of saponin containing no isoflavone from soybean embro bud - Google Patents

Abstract

PURPOSE:To obtain saponin of high purity without containing isoflavones on an industrial scale, by adding water to an extract concentrate of soybean embryo buds with a lower alcohol, adsorbing the extract on an adsorbent, eluting the adsorbed extract stepwise with a hydrophilic organic solvent and purifying the eluate by gel filtration, etc. CONSTITUTION:Soybean embryo buds are extracted with a lower alcohol and concentrated, and water is then added thereto. The resultant extract is then adsorbed on an adsorbent, e.g. HP-20, HP-40, etc., and isoflavones are mostly washed and removed with 10-40% methanol, ethanol, acetone, etc. The adsorbed extract is subsequently eluted with 70-80% methanol, ethanol, etc., to give an eluate, which is then purified by gel filtration and partition chromatography to afford the aimed saponin containing no isoflavones. Thereby, the isoflavones giving side effect to the human body are removed from the soybean embryo buds with a higher saponin content than soybean flour and the aimed saponin of high purity can be obtained in large amounts industrially.

Description

【発明の詳細な説明】 本発明は、大豆粉よシサポニン含量が多い大豆胚芽に着
目し、しかも人体に副作用を与えるインフラＲン類を排
除できる新知見の精製法に由来する大豆胚芽よりサポニ
ンを工業的に取得可能な新規のサポニン製造法に関する
ものである。Detailed Description of the Invention The present invention focuses on soybean germ, which has a higher content of shisaponin than soybean flour, and moreover, produces saponin from soybean germ derived from a new purification method that can eliminate infrared substances that cause side effects to the human body. This invention relates to a new industrially obtainable saponin production method.

大豆胚芽は強い苦味成分があるため、豆乳製造工程の初
期段階において機械的ふるい分けがなされ、利用価値の
低い産業廃棄物として処理されてきた。Because soybean germ has a strong bitter taste, it has been mechanically sieved in the early stages of the soymilk manufacturing process and has been treated as industrial waste with low utility value.

本発明者は、一般にサポニン原料として周知の大豆粉よ
シ数倍のサポニン含量を有する大豆胚芽から、来雑物を
有しないサポニン抽出法について鋭意研究の結果、ここ
に高純度サポニンを工業的に大量取得する製造法を完成
するに至った。As a result of intensive research into a method for extracting saponin free of impurities from soybean germ, which has a saponin content several times higher than that of soybean flour, which is generally known as a raw material for saponin, the present inventor has developed an industrial method for extracting high-purity saponin. We have completed a manufacturing method to obtain large quantities.

従来公知の方法によれば、サポニン類は、その精製単離
にあたシ、アルコール抽出後、シリカグゝルカラムクロ
マトグラフィー、もしくはカウンターカレントディスト
リビー−ターなどが用いられておシ、サポニンの大量単
離には、困難な点が多かった。また、大豆成分の中でサ
ポニンの精製において最も問題となるのは、ダイゼン、
デニステイン等のイソフラデンの存在である。これらは
、溶媒転溶による抽出工程でサポニンと分離することが
困難であシ、更に、これらインフラ？ン類は、女性ホル
モン様作用を有し、哺乳類に不妊症をはじめとする副作
用を惹起せしめることが知られている。According to conventionally known methods, saponins are purified and isolated using silica gel column chromatography, counter current distributor, etc. after alcohol extraction. There were many difficulties in isolation. Also, among the soybean ingredients, the most problematic in refining saponin is daizen,
The presence of isofradenes such as denistein. These are difficult to separate from saponin in the extraction process by solvent transfer, and furthermore, these infrastructures are difficult to separate. It is known that sterilants have female hormone-like effects and cause side effects such as infertility in mammals.

従って、サポニンを長期間にわたシ、よシ安全に服用す
るためには、製造工程中において、インフラ？ン類との
完全分離が必須であると考えられる。Therefore, in order to safely take saponin over a long period of time, it is necessary to have sufficient infrastructure during the manufacturing process. It is considered that complete separation from other substances is essential.

即ち、本発明は、大豆胚芽に、低級アルコール、例工ば
メタノール、エタノール、含水ブタノール等、捷たは其
の酸性含水物を加え、抽出液を濃縮後に水を加え、ＨＰ
−２０，４０％ＸＡＤ　−１、２。That is, in the present invention, a lower alcohol such as methanol, ethanol, aqueous butanol, etc., or an acidic hydrate thereof is added to soybean germ, the extract is concentrated, water is added, and HP
-20,40%XAD-1,2.

４．７，８、カラムライト等の吸着樹脂、または活性炭
に吸着させ、１０〜４０％メタノール、エタノール、ア
セトン等で洗浄し、次いで７０〜８０係メタノール、エ
タノール、アセトン等によシ溶出するか、もしくは、分
子ふるいＬＨ−２０−、ＬＨ−６０を用いたメタノール
、エタノール、アセトンなどの親水性溶媒および其の含
水物を用いたクロマトグラフィー、もしくは、セライト
などの担体を用いた分配クロマトグラフィーによシ、精
製を行うことを特徴とする。4.7, 8. Adsorb onto adsorption resin such as Columnite or activated carbon, wash with 10-40% methanol, ethanol, acetone, etc., and then elute with 70-80% methanol, ethanol, acetone, etc. Alternatively, chromatography using molecular sieves LH-20- and LH-60 with hydrophilic solvents such as methanol, ethanol, and acetone and their hydrates, or partition chromatography using a carrier such as Celite. It is characterized by refining.

今回、発明者はＨＰ−２０など吸着樹脂から親水性の有
機溶媒によ多段階的に溶出する方法およびＬＨ−２０な
どｒル濾過剤によるカラムクロマトグラフィーとを組み
合わせることによシ、インフラぎンとす？ニンを完全に
分離することに成功し、しかも、本発明による製法は、
大量処理が可能な精製サポニンを工業的に採取し得るも
のである。This time, the inventors have developed an infragravity method by combining a multi-step elution method from an adsorption resin such as HP-20 with a hydrophilic organic solvent and column chromatography using an IR filtration agent such as LH-20. And? In addition, the production method according to the present invention has succeeded in completely separating Nin.
Purified saponin that can be processed in large quantities can be industrially harvested.

また、サポニン抽出原料として大豆胚芽を採用したこと
によシ、サポニン収量は、大豆粉の場合に比し、６倍以
上に達する。Furthermore, by using soybean germ as the raw material for saponin extraction, the yield of saponin is more than six times that of soybean flour.

しかも、大豆全体より得られたサポニン群と大豆胚芽よ
シ得られたサポニン群では其の組成比を全く異にしてお
り（第１図、第２図参照）、従って薬理学的にも両者は
質的もしくは量的に特異な作用が期待される。Moreover, the saponins obtained from whole soybeans and the saponins obtained from soybean germ have completely different composition ratios (see Figures 1 and 2), and therefore pharmacologically speaking, they are different from each other. A qualitatively or quantitatively unique effect is expected.

実施例１ 大豆胚芽１　ｋｇを０．００１Ｎ−塩酸・９０係メタノ
ール３Ｌで抽出し、ヘキサンＩＬを加え３回脱脂した。Example 1 1 kg of soybean germ was extracted with 3 L of 0.001 N hydrochloric acid and 90% methanol, and defatted three times by adding hexane IL.

抽出液の溶媒を減圧下で留去した後、水で２Ｌとし吸着
樹脂ＨＰ−２０に付した。２０係及び７０％アセトンで
溶出し、後者を減圧濃縮して、２４．３８，９の粗粉末
を得た。そのうち５．５０ｇを９９幅メタノール２０Ｔ
ＩＬｌに溶解し、５ｅｐｈａｄｅｘ　Ｉ、Ｈ−２０デル
濾過クロマトグラフイーに付した。After the solvent of the extract was distilled off under reduced pressure, the volume was made up to 2 L with water and applied to adsorption resin HP-20. Elution was carried out with 20% and 70% acetone, and the latter was concentrated under reduced pressure to obtain a crude powder of 24.38.9. Of that, 5.50g is 99 width methanol 20T
It was dissolved in IL1 and subjected to 5ephadex I, H-20 Delfiltration chromatography.

溶出液は薄層クロマトグラフィー〔担体−シリカデル６
０Ｆ２５４、展開溶媒＝ｎ−ブタノール・酢酸・水（３
：１：１　）、発色剤＝１チ硫酸第２セリウム１０％硫
酸溶液噴霧〕で検出し、Ｒｆｏ、４に出現するサポニン
の単一スポットを確認した。このフラクションを減圧し
て３．１（Ｈｉｔの結晶を得だ。収率１．３８係。The eluate was subjected to thin layer chromatography [support: silica del 6].
0F254, developing solvent = n-butanol/acetic acid/water (3
:1:1), color former = 1 ceric sulfate, 10% sulfuric acid solution spray], and a single spot of saponin appearing at Rfo, 4 was confirmed. This fraction was depressurized and crystals of 3.1 (Hit) were obtained. Yield: 1.38%.

実施例２　　′ 実施例１と同様に処理して得られた大豆胚芽よシのＨＰ
−２０・アセトン抽出物２３．４８ｇの粗粉末を得た。Example 2 ' HP of soybean germ obtained by the same treatment as in Example 1
23.48 g of crude powder of -20.acetone extract was obtained.

そのうち１．５ｇをｎ−ブタノールに溶解してセライト
分配クロマトグラフィーを行った。1.5 g of it was dissolved in n-butanol and subjected to celite partition chromatography.

溶出溶媒はブタノールを用い展開した。The elution solvent was developed using butanol.

実施例１と同様に薄層クロマトグラフィーにより、Ｒｆ
約０．４に出現するサポニンを確認した。このフラクシ
ョンを減圧濃縮し２．９８ｇの結晶を得た。By thin layer chromatography as in Example 1, Rf
Saponin appearing at approximately 0.4 was confirmed. This fraction was concentrated under reduced pressure to obtain 2.98 g of crystals.

収率１．４係。Yield: 1.4.

【図面の簡単な説明】[Brief explanation of the drawing]

第１図は従来公知の方法により大豆全体より得られたサ
ポニン群の組成を示す分析グラフ、第２図は本発明方法
により大豆胚芽より得られたサポニン群の組成を示す分
析グラフである。 第　　１　　図 第２１７１FIG. 1 is an analytical graph showing the composition of a saponin group obtained from whole soybean by a conventionally known method, and FIG. 2 is an analytical graph showing the composition of a saponin group obtained from soybean germ by the method of the present invention. Figure 1 Figure 2171

Claims (1)

【特許請求の範囲】[Claims] 大豆胚芽を低級アルコールで抽出し、濃縮後に水を加え
吸着剤に吸着させた後、低濃度の低級アルコールで大部
分のイソフラボン類を洗浄除去し、続いて吸着体を高濃
度の低級アルコール又はアセトンで溶出した液について
ゲルろ過又は分配クロマトグラフィーを用いて精製する
ことを特徴とする大豆胚芽よりのイソフラボン類を含有
しないサポニンの製造法。Soybean germ is extracted with a lower alcohol, water is added after concentration, and the adsorbent is adsorbed. Most of the isoflavones are washed away with a low concentration of lower alcohol, and then the adsorbent is extracted with a high concentration of lower alcohol or acetone. 1. A method for producing saponin from soybean germ that does not contain isoflavones, the method comprising purifying the eluted solution using gel filtration or partition chromatography.