JP2002371032A - 3,5-diprenyl-4-hydroxycinnamic acid for standard sample and method for producing the same - Google Patents
3,5-diprenyl-4-hydroxycinnamic acid for standard sample and method for producing the sameInfo
- Publication number
- JP2002371032A JP2002371032A JP2001181478A JP2001181478A JP2002371032A JP 2002371032 A JP2002371032 A JP 2002371032A JP 2001181478 A JP2001181478 A JP 2001181478A JP 2001181478 A JP2001181478 A JP 2001181478A JP 2002371032 A JP2002371032 A JP 2002371032A
- Authority
- JP
- Japan
- Prior art keywords
- diprenyl
- hydroxycinnamic acid
- propolis
- producing
- alcohol
- 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.)
- Granted
Links
- KABCFARPAMSXCC-UHFFFAOYSA-N artepillin C Natural products CC(C)=CCC1=CC(C=CC(O)=O)=CC(CC=C(C)C)=C1O KABCFARPAMSXCC-UHFFFAOYSA-N 0.000 title claims abstract description 62
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 6
- 241000241413 Propolis Species 0.000 claims abstract description 28
- 229940069949 propolis Drugs 0.000 claims abstract description 28
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 9
- 238000004007 reversed phase HPLC Methods 0.000 claims abstract description 6
- 238000010898 silica gel chromatography Methods 0.000 claims description 3
- NGSWKAQJJWESNS-UHFFFAOYSA-N 4-coumaric acid Chemical compound OC(=O)C=CC1=CC=C(O)C=C1 NGSWKAQJJWESNS-UHFFFAOYSA-N 0.000 claims 2
- 229930005346 hydroxycinnamic acid Natural products 0.000 claims 1
- 235000010359 hydroxycinnamic acids Nutrition 0.000 claims 1
- NGSWKAQJJWESNS-ZZXKWVIFSA-N trans-4-coumaric acid Chemical compound OC(=O)\C=C\C1=CC=C(O)C=C1 NGSWKAQJJWESNS-ZZXKWVIFSA-N 0.000 claims 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 abstract description 8
- 238000004458 analytical method Methods 0.000 abstract description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 5
- 238000004587 chromatography analysis Methods 0.000 abstract description 2
- 239000000377 silicon dioxide Substances 0.000 abstract description 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 24
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 6
- 239000003480 eluent Substances 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 238000004949 mass spectrometry Methods 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 239000000741 silica gel Substances 0.000 description 4
- 229910002027 silica gel Inorganic materials 0.000 description 4
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 238000004440 column chromatography Methods 0.000 description 3
- 239000000470 constituent Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 241000257303 Hymenoptera Species 0.000 description 2
- 230000001093 anti-cancer Effects 0.000 description 2
- 125000004177 diethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 239000012044 organic layer Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N Acrylic acid Chemical compound OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- PPWHTZKZQNXVAE-UHFFFAOYSA-N Tetracaine hydrochloride Chemical compound Cl.CCCCNC1=CC=C(C(=O)OCCN(C)C)C=C1 PPWHTZKZQNXVAE-UHFFFAOYSA-N 0.000 description 1
- 238000000862 absorption spectrum Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000013985 cinnamic acid Nutrition 0.000 description 1
- WBYWAXJHAXSJNI-UHFFFAOYSA-N cinnamic acid group Chemical class C(C=CC1=CC=CC=C1)(=O)O WBYWAXJHAXSJNI-UHFFFAOYSA-N 0.000 description 1
- 238000002451 electron ionisation mass spectrometry Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- SVWLIIFHXFGESG-UHFFFAOYSA-N formic acid;methanol Chemical compound OC.OC=O SVWLIIFHXFGESG-UHFFFAOYSA-N 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000011785 micronutrient Substances 0.000 description 1
- 235000013369 micronutrients Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Landscapes
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、プロポリスから
３，５−ジプレニル−４−ヒドロキシ桂皮酸 ( ３，５
−diprenyl−４−hydroxycinnamic acid )を製造する方
法に関する。The present invention relates to a process for producing 3,5-diprenyl-4-hydroxycinnamic acid (3,5) from propolis.
-Diprenyl-4-hydroxycinnamic acid).
【０００２】[0002]
【従来の技術】プロポリスは、一般にハチヤニとも呼ば
れ、ミツバチが花や樹脂のヤニ状成分をかみ続けること
により、各構成成分とミツバチの唾液中の酵素とが混ぜ
合わされて生じる物質である。このプロポリスには、フ
ラボノイド類、桂皮酸類、テルペノイド、ミネラル、ビ
タミンなどの微量栄養素を含有している為、健康の増進
に寄与したり、治療的効果（殺菌、抗炎症など）を有す
るものと期待されている。2. Description of the Related Art Propolis, which is also commonly referred to as bee crab, is a substance that is produced when bees continue to bite the flower-like or resin-like constituents of a flower, thereby mixing each constituent with enzymes in the saliva of the bees. Since this propolis contains micronutrients such as flavonoids, cinnamic acids, terpenoids, minerals, and vitamins, it is expected to contribute to health promotion and have therapeutic effects (bactericidal, anti-inflammatory, etc.) Have been.
【０００３】プロポリスの成分は非常に多様であり、ま
た、産地によって由来植物が異なるために成分も産地に
よりかなり異なってくる。同じブラジル産プロポリスで
もその成分は多少異なるが、地域によりある程度の似通
った成分のプロポリスが採取される傾向が有るとされて
いる。[0003] The components of propolis are very diverse, and the components vary considerably depending on the place of origin, because the origin plants differ depending on the place of origin. Although the components of Brazilian propolis are somewhat different, it is said that there is a tendency for propolis to be collected to a certain degree depending on the region.
【０００４】近年、プロポリス、特に、ブラジル産プロ
ポリスでの有効成分の一つとして、３，５−ジプレニル
−４−ヒドロキシ桂皮酸（図１参照）の有用性が注目さ
れ、抗がん剤や殺菌剤等としての研究に関する報告が、
例えば、Kimoto TらのAnticancer Research 21:221-228
(2001)、Kimoto TらのPathol Int. 2000 Sep;50(9):679
-89. 、Kimoto TらのCancer Detect Prev. 1998;22(6):
506-15. 、Matsuno TらのAnticancer Res. 1997 Sep-Oc
t;17(5A):3565-8. 、Kimoto TらのGan To Kagaku Ryoh
o. 1996 Nov;23(13):1855-9. とAga Hらの(1994) Biote
ch.Biochem. 58, 945-946.等に見られる。[0004] In recent years, the usefulness of 3,5-diprenyl-4-hydroxycinnamic acid (see Fig. 1) as one of the active ingredients in propolis, particularly in Brazilian propolis, has been attracting attention, and it has attracted attention as an anticancer agent and a bactericidal agent. Reports on research as agents, etc.
For example, Kimoto T et al. Anticancer Research 21: 221-228
(2001), Kimoto T et al., Pathol Int. 2000 Sep; 50 (9): 679.
-89., Kimoto T et al., Cancer Detect Prev. 1998; 22 (6):
506-15., Matsuno T et al. Anticancer Res. 1997 Sep-Oc
t; 17 (5A): 3565-8., Kimoto T et al., Gan To Kagaku Ryoh
o. 1996 Nov; 23 (13): 1855-9. and Aga H et al. (1994) Biote.
ch. Biochem. 58, 945-946.
【０００５】このような３，５−ジプレニル−４−ヒド
ロキシ桂皮酸を含有するプロポリス原料の品質を評価し
管理するには原料に応じて異なる３，５−ジプレニル−
４−ヒドロキシ含有量を分析、定量することが必要であ
り、そのためには、３，５−ジプレニル−４−ヒドロキ
シ桂皮酸の標準品が必要である。しかしながら、３，５
−ジプレニル−４−ヒドロキシ桂皮酸の標準品は、試薬
として市販されていない。その理由の一つは、分析用標
準品として好適な高純度の３，５−ジプレニル−４−ヒ
ドロキシ桂皮酸を簡単に得ることができるような製法が
確立されていないことによるものと考えられる。例え
ば、特開昭６０−１６３８４１には、３，５−ジプレニ
ル−４−ヒドロキシ桂皮酸を含むプロペニック酸の合成
法が開示されているが、多くの複雑な工程を要し、標準
品に用いるような多量の３，５−ジプレニル−４−ヒド
ロキシ桂皮酸を製造するには適していない。[0005] In order to evaluate and control the quality of such a propolis raw material containing 3,5-diprenyl-4-hydroxycinnamic acid, a different 3,5-diprenyl-derivative depends on the raw material.
It is necessary to analyze and quantify the 4-hydroxy content, and for that purpose, a standard 3,5-diprenyl-4-hydroxycinnamic acid is required. However, 3,5
-Diprenyl-4-hydroxycinnamic acid standard is not commercially available as a reagent. One of the reasons is considered to be that a production method capable of easily obtaining high-purity 3,5-diprenyl-4-hydroxycinnamic acid suitable as an analytical standard has not been established. For example, JP-A-60-163841 discloses a method for synthesizing propenic acid containing 3,5-diprenyl-4-hydroxycinnamic acid, but requires a number of complicated steps, and may be used as a standard product. It is not suitable for producing very large amounts of 3,5-diprenyl-4-hydroxycinnamic acid.
【０００６】[0006]
【発明が解決しようとする課題】本発明の目的は、３，
５−ジプレニル−４−ヒドロキシ桂皮酸含有量の分析を
行う為に、標準品としての３，５−ジプレニル−４−ヒ
ドロキシ桂皮酸を製造することのできる新しい方法を開
発することにある。SUMMARY OF THE INVENTION The object of the present invention is to
In order to analyze the content of 5-diprenyl-4-hydroxycinnamic acid, it is an object of the present invention to develop a new method capable of producing 3,5-diprenyl-4-hydroxycinnamic acid as a standard.
【０００７】[0007]
【課題を解決するための手段】本発明者は、プロポリス
そのものから３，５−ジプレニル−４−ヒドロキシ桂皮
酸を高純度且つ比較的簡単に抽出、精製し得る方法を確
立し、上記の目的を達成したものである。Means for Solving the Problems The present inventors have established a method for extracting and purifying 3,5-diprenyl-4-hydroxycinnamic acid from propolis itself with high purity and relatively easily. It has been achieved.
【０００８】かくして、本発明に従えば、プロポリスを
アルコールまたは含水アルコールで抽出して得られる抽
出液をシリカクロマトグラフィーおよび逆相ＨＰＬＣに
より精製する工程を含む３，５−ジプレニル−４−ヒド
ロキシ桂皮酸の製造方法が提供される。さらに、本発明
は、別の視点として、上記のようにプロポリスから製造
され３，５−ジプレニル−４−ヒドロキシ桂皮酸を分
析、特にプロポリス中の３，５−ジプレニル−４−ヒド
ロキシ桂皮酸を分析するための標準品として用いられる
３，５−ジプレニル−４−ヒドロキシ桂皮酸を提供す
る。Thus, according to the present invention, 3,5-diprenyl-4-hydroxycinnamic acid comprising a step of purifying an extract obtained by extracting propolis with alcohol or hydrous alcohol by silica chromatography and reverse phase HPLC. Is provided. Further, the present invention provides, as another aspect, the analysis of 3,5-diprenyl-4-hydroxycinnamic acid produced from propolis as described above, and in particular, the analysis of 3,5-diprenyl-4-hydroxycinnamic acid in propolis. 3,5-diprenyl-4-hydroxycinnamic acid, which is used as a standard for the production of
【０００９】[0009]
【発明の実施の形態】本発明の方法は、多様な構成成分
から成るプロポリスから、３，５−ジプレニル−４−ヒ
ドロキシ桂皮酸を効率的に抽出、製製することのできる
最適な工程を見出したことに基づくものであり、その基
本的な実施工程として、プロポリスをアルコールまたは
含水アルコールで抽出する工程、抽出液をシリカゲルを
用いるカラムクロマトグラフィーで精製する工程、およ
び逆相ＨＰＬＣ（高速液体クロマトグラフィー）により
精製する工程を含む。BEST MODE FOR CARRYING OUT THE INVENTION The method of the present invention finds an optimum step for efficiently extracting and producing 3,5-diprenyl-4-hydroxycinnamic acid from propolis composed of various constituents. As a basic implementation step, the steps of extracting propolis with alcohol or hydrous alcohol, purifying the extract by column chromatography using silica gel, and reverse phase HPLC (high performance liquid chromatography) )).
【００１０】原料となるプロポリスの例としては、３，
５−ジプレニル−４−ヒドロキシ桂皮酸を比較的多量に
含有する地域由来のブラジル産プロポリス原塊を用いる
のが好ましい。アルコールまたは含水アルコールによる
抽出工程に用いられる好ましいアルコールとしては、一
般に、メタノールであるが、エタノールを用いることも
できる。この工程によってプロポリス原料中の主要な構
成成分であるメタノール可溶成分が抽出される。Examples of propolis as a raw material include 3,
It is preferable to use an original Brazilian propolis mass derived from a region containing a relatively large amount of 5-diprenyl-4-hydroxycinnamic acid. The preferred alcohol used in the extraction step with alcohol or hydrous alcohol is generally methanol, but ethanol can also be used. By this step, a methanol-soluble component, which is a main component in the propolis raw material, is extracted.
【００１１】アルコールによる抽出液（抽出したアルコ
ール溶液）は、シリカゲルを用いるカラムクロマトグラ
フィー工程に供されて、極性成分（高極性成分および低
極性成分）が除去される。この際、カラムクロマトグラ
フィーへの負荷を軽減するために、抽出したアルコール
溶液は、予め、水を加えることにより不純物を沈殿物と
して除き、更に水洗浄することを含む前処理を行うこと
が好ましい。The alcohol extract (the extracted alcohol solution) is subjected to a column chromatography step using silica gel to remove polar components (high and low polar components). At this time, in order to reduce the load on the column chromatography, it is preferable that the extracted alcohol solution is preliminarily subjected to a pretreatment including adding water to remove impurities as precipitates and further washing with water.
【００１２】シリカゲルカラムクロマトグラフィー工程
は、例えば、クロロホルム／メタノール混合溶媒を溶離
液として行うのが好ましい。シリカゲルカラムクロマト
グラフィーで分離された画分は、濃縮後、例えばメタノ
ールで希釈し、不溶物を例えば遠心分離により除去して
逆相ＨＰＬＣ用サンプルを調製する。The silica gel column chromatography step is preferably performed, for example, using a mixed solvent of chloroform / methanol as an eluent. The fraction separated by silica gel column chromatography is concentrated and then diluted with, for example, methanol, and the insoluble matter is removed by, for example, centrifugation to prepare a sample for reversed phase HPLC.
【００１３】逆相ＨＰＬＣの固定相（カラム）として好
ましい例は、ＯＤＳであり、例えば、ギ酸／メタノール
を溶離液（移動相）として液体クロマトグラフィー操作
を行う。目的の画分はジエチルエーテル可溶分として集
め、濃縮した後、残渣を例えばヘキサンで洗浄し、乾燥
することにより３，５−ジプレニル−４−ヒドロキシ桂
皮酸が得られる。A preferred example of the stationary phase (column) of the reversed phase HPLC is ODS. For example, liquid chromatography is carried out using formic acid / methanol as an eluent (mobile phase). The target fraction is collected as a diethyl ether-soluble component, concentrated, and the residue is washed with, for example, hexane, and dried to obtain 3,5-diprenyl-4-hydroxycinnamic acid.
【００１４】[0014]
【実施例】次に、本発明による製造法の好適な具体例を
示すが、本発明はこれに限定されるものではない。ブラ
ジル産プロポリス原塊１５０ｇにメタノール４５０ｍｌ
を加え、室温で攪拌してメタノール可溶成分を、抽出し
た。これをろ過することにより抽出液を得た。抽出液に
は１／１０量の水を加え、沈殿物を遠心分離により残さ
として除いた。上清に等量の酢酸エチルを加え、１／２
量の水で有機層を洗浄した。回収した有機層は、濃縮
後、メタノール：クロロホルムにて再溶解した（固形分
２０．９ｇ）。固形分２０．２ｇに相当する溶液をシ
リカゲルカラム (シリカゲル２００ｇ、５０ｍｍφ)に
負荷し、クロロホルム／メタノール（９：１）を溶離液
として分離した。黒褐色の部分の画分を集めて濃縮し、
黒褐色オイル２２．４ｇを得た。メタノール希釈し、不
溶物を遠沈除去し、ＨＰＬＣ分取サンプルとした。逆相
(ＯＤＳ)カラムにて、５％ギ酸−メタノール（３５ｖｏ
ｌ／６５ｖｏｌ）を溶離液として分離・分画し、目的画
分を集め濃縮した後、ジエチルエーテル可溶分を集め、
再度濃縮し、残さをヘキサンで洗浄し、減圧乾燥するこ
とにより黄白色アモルファス状の固体０．１３４ｇを得
た（収率０．０９％）。これは、ＨＰＬＣによる純度分
析により３，５−ジプレニル−４−ヒドロキシ桂皮酸を
９６．６％含むことが確認された（図２および図３参
照）。またその一部を用い、以下のように、1Ｈ−ＮＭ
Ｒ（ＣＤＣｌ3／ＴＭＳ）およびＭＳ（質量分析）によ
り３，５−ジプレニル−４−ヒドロキシ桂皮酸であるこ
とを確認した。 ＭＳ，ｍ／ｅ；３００[Ｍ]−，２８５[Ｍ−ＣＨ3]＋
（ＥＩＭＳ法、７０ｅＶ）。1 Ｈ−ＮＭＲ(３００ＭＨｚ，ＣＤＣｌ3)，内部標準 Ｔ
ＭＳ，δ(ｐｐｍ)；７．６７(１Ｈ，ｄ，Ｊ＝１５．９
Ｈｚ)、７．１５(２Ｈ，ｂｒｓ)、６．２３(１Ｈ，ｄ，
Ｊ＝１５．９Ｈｚ)、５．２８(２Ｈ，ｂｒｔ)、３．３
３(４Ｈ，ｍ)、１．７８(１２Ｈ，ｓ)。Next, preferred specific examples of the production method according to the present invention will be described, but the present invention is not limited thereto. 450ml of methanol in 150g of original Brazilian propolis mass
And stirred at room temperature to extract methanol-soluble components. This was filtered to obtain an extract. 1/10 volume of water was added to the extract, and the precipitate was removed by centrifugation. Add an equal volume of ethyl acetate to the supernatant,
The organic layer was washed with an amount of water. The collected organic layer was concentrated and then redissolved in methanol: chloroform (solid content 20.9 g). A solution corresponding to a solid content of 20.2 g was loaded on a silica gel column (silica gel 200 g, 50 mmφ) and separated using chloroform / methanol (9: 1) as an eluent. The fraction of the black-brown part is collected and concentrated,
22.4 g of a dark brown oil were obtained. The mixture was diluted with methanol, and the insolubles were removed by centrifugation. Reversed phase
(ODS) column, 5% formic acid-methanol (35 vo
1/65 vol) as an eluent, separating and fractionating the target fraction, concentrating and collecting the diethyl ether-soluble matter,
The solution was concentrated again, and the residue was washed with hexane and dried under reduced pressure to obtain 0.134 g of a yellow-white amorphous solid (yield 0.09%). This was confirmed to contain 96.6% of 3,5-diprenyl-4-hydroxycinnamic acid by purity analysis by HPLC (see FIGS. 2 and 3). Further, using a part thereof, 1 H-NM
It was confirmed by R (CDCl 3 / TMS) and MS (mass spectrometry) that it was 3,5-diprenyl-4-hydroxycinnamic acid. MS, m / e; 300 [M] − , 285 [M-CH 3] +
(EIMS method, 70 eV). 1 H-NMR (300 MHz, CDCl 3 ), internal standard T
MS, δ (ppm); 7.67 (1H, d, J = 15.9)
Hz), 7.15 (2H, brs), 6.23 (1H, d,
J = 15.9 Hz), 5.28 (2H, brt), 3.3
3 (4H, m), 1.78 (12H, s).
【００１５】[0015]
【発明の効果】本発明に従えば、プロポリスを出発原料
として比較的簡単な工程により高純度の３，５−ジプレ
ニル−４−ヒドロキシ桂皮酸を製造することができる。
本発明によって得られる３，５−ジプレニル−４−ヒド
ロキシ桂皮酸は、プロポリス原塊やプロポリス製品中の
３，５−ジプレニル−４−ヒドロキシ桂皮酸含有量を分
析するための標準品として用いられることができ、それ
らの安定した品質管理および品質向上に資することがで
きる。According to the present invention, high purity 3,5-diprenyl-4-hydroxycinnamic acid can be produced by a relatively simple process using propolis as a starting material.
The 3,5-diprenyl-4-hydroxycinnamic acid obtained according to the present invention is used as a standard product for analyzing the content of 3,5-diprenyl-4-hydroxycinnamic acid in the original propolis mass and propolis products. And contribute to their stable quality control and quality improvement.
【図１】３，５−ジプレニル−４−ヒドロキシ桂皮酸の
構造式である。FIG. 1 is a structural formula of 3,5-diprenyl-4-hydroxycinnamic acid.
【図２】３，５−ジプレニル−４−ヒドロキシ桂皮酸純
度の確認の為に行った、高速液体クロマトグラフィーの
クロマトグラムである。FIG. 2 is a chromatogram of high-performance liquid chromatography performed for confirming the purity of 3,5-diprenyl-4-hydroxycinnamic acid.
【図３】図２での高速液体クロマトグラフィーのクロマ
トグラムにおける３，５−ジプレニル−４−ヒドロキシ
桂皮酸のピーク（図２の保持期間１０．６０分）の紫外
吸収スペクトルである。FIG. 3 is an ultraviolet absorption spectrum of the peak of 3,5-diprenyl-4-hydroxycinnamic acid (retention period of 10.60 minutes in FIG. 2) in the chromatogram of the high performance liquid chromatography in FIG.
Claims (3)
コールで抽出して得られる抽出液をシリカゲルカラムク
ロマトグラフィーおよび逆相ＨＰＬＣにより精製する工
程を含むことを特徴とする３，５−ジプレニル−４−ヒ
ドロキシ桂皮酸の製造方法。1. A process comprising purifying an extract obtained by extracting propolis with an alcohol or a hydrous alcohol by silica gel column chromatography and reverse phase HPLC, and comprising 3,5-diprenyl-4-hydroxycinnamic acid. Manufacturing method.
ニル−４−ヒドロキシ桂皮酸を分析するための標準品と
して用いられることを特徴とする３，５−ジプレニル−
４−ヒドロキシ桂皮酸。2. A 3,5-diprenyl-produced from propolis, which is used as a standard for analyzing 3,5-diprenyl-4-hydroxycinnamic acid.
4-hydroxycinnamic acid.
−ヒドロキシ桂皮酸を分析するための標準品として用い
られることを特徴とする請求項２の３，５−ジプレニル
−４−ヒドロキシ桂皮酸。3. The 3,5-diprenyl-4 in propolis
3. The 3,5-diprenyl-4-hydroxycinnamic acid according to claim 2, which is used as a standard for analyzing hydroxycinnamic acid.
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|---|---|---|---|
| JP2001181478A JP4822616B2 (en) | 2001-06-15 | 2001-06-15 | 3,5-diprenyl-4-hydroxycinnamic acid for standard products and method for producing the same |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103267823A (en) * | 2013-05-27 | 2013-08-28 | 杭州蜂之语蜂业股份有限公司 | Method for Determination of Artepillin C Content in Propolis by High Performance Liquid Chromatography |
| CN109704951A (en) * | 2019-03-01 | 2019-05-03 | 汤臣倍健股份有限公司 | A kind of extracting method of Ah Ti woods C |
| CN114573446A (en) * | 2020-12-02 | 2022-06-03 | 中国科学院大连化学物理研究所 | Method for preparing protocatechuic acid from phellinus igniarius medicinal material |
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|---|---|---|---|---|
| JPS60163841A (en) * | 1984-02-06 | 1985-08-26 | Res Inst For Prod Dev | 3-(4-hydroxyphenyl)-2(E)-propenic acid |
| JPH07330596A (en) * | 1994-06-01 | 1995-12-19 | Eiken Chem Co Ltd | Antitumor agent |
| JPH09151131A (en) * | 1995-09-29 | 1997-06-10 | Hayashibara Biochem Lab Inc | Medicine for methicillin-resistant staphylococcus aureus infectious disease |
| JP2002332257A (en) * | 2001-05-11 | 2002-11-22 | Biomedeikusu:Kk | Method for extracting and purifying 3-[4-hydroxy-3, 5-bis (3-methyl-2-butenyl) phenyl]-2-propenoic acid from propolis |
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2001
- 2001-06-15 JP JP2001181478A patent/JP4822616B2/en not_active Expired - Lifetime
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS60163841A (en) * | 1984-02-06 | 1985-08-26 | Res Inst For Prod Dev | 3-(4-hydroxyphenyl)-2(E)-propenic acid |
| JPH07330596A (en) * | 1994-06-01 | 1995-12-19 | Eiken Chem Co Ltd | Antitumor agent |
| JPH09151131A (en) * | 1995-09-29 | 1997-06-10 | Hayashibara Biochem Lab Inc | Medicine for methicillin-resistant staphylococcus aureus infectious disease |
| JP2002332257A (en) * | 2001-05-11 | 2002-11-22 | Biomedeikusu:Kk | Method for extracting and purifying 3-[4-hydroxy-3, 5-bis (3-methyl-2-butenyl) phenyl]-2-propenoic acid from propolis |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103267823A (en) * | 2013-05-27 | 2013-08-28 | 杭州蜂之语蜂业股份有限公司 | Method for Determination of Artepillin C Content in Propolis by High Performance Liquid Chromatography |
| CN109704951A (en) * | 2019-03-01 | 2019-05-03 | 汤臣倍健股份有限公司 | A kind of extracting method of Ah Ti woods C |
| CN109704951B (en) * | 2019-03-01 | 2022-02-18 | 汤臣倍健股份有限公司 | Extraction method of aspirin C |
| CN114573446A (en) * | 2020-12-02 | 2022-06-03 | 中国科学院大连化学物理研究所 | Method for preparing protocatechuic acid from phellinus igniarius medicinal material |
| CN114573446B (en) * | 2020-12-02 | 2023-05-30 | 中国科学院大连化学物理研究所 | Method for preparing protocatechuic acid from phellinus linteus medicinal material |
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|---|---|
| JP4822616B2 (en) | 2011-11-24 |
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