JPS63278971A - Removal of acid from acidic anthocyanin pigment solution - Google Patents
Removal of acid from acidic anthocyanin pigment solutionInfo
- Publication number
- JPS63278971A JPS63278971A JP62113201A JP11320187A JPS63278971A JP S63278971 A JPS63278971 A JP S63278971A JP 62113201 A JP62113201 A JP 62113201A JP 11320187 A JP11320187 A JP 11320187A JP S63278971 A JPS63278971 A JP S63278971A
- Authority
- JP
- Japan
- Prior art keywords
- dye
- acid
- pigment
- anthocyanin
- acidic
- 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
- 229930002877 anthocyanin Natural products 0.000 title claims abstract description 29
- 235000010208 anthocyanin Nutrition 0.000 title claims abstract description 29
- 239000004410 anthocyanin Substances 0.000 title claims abstract description 29
- 150000004636 anthocyanins Chemical class 0.000 title claims abstract description 29
- 239000000049 pigment Substances 0.000 title claims abstract description 28
- 230000002378 acidificating effect Effects 0.000 title claims abstract description 10
- 239000002253 acid Substances 0.000 title abstract description 19
- 229920005989 resin Polymers 0.000 claims abstract description 21
- 239000011347 resin Substances 0.000 claims abstract description 21
- 239000003480 eluent Substances 0.000 claims abstract description 14
- 239000000243 solution Substances 0.000 claims abstract description 8
- 239000003463 adsorbent Substances 0.000 claims abstract description 6
- 239000007864 aqueous solution Substances 0.000 claims abstract description 5
- 238000000034 method Methods 0.000 claims description 8
- 238000005406 washing Methods 0.000 claims description 7
- 239000000758 substrate Substances 0.000 claims 1
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 abstract description 31
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 abstract description 6
- 229920001429 chelating resin Polymers 0.000 abstract description 3
- 230000000274 adsorptive effect Effects 0.000 abstract 2
- 238000005192 partition Methods 0.000 abstract 2
- 230000002745 absorbent Effects 0.000 abstract 1
- 239000002250 absorbent Substances 0.000 abstract 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 21
- VEVZSMAEJFVWIL-UHFFFAOYSA-O cyanidin cation Chemical compound [O+]=1C2=CC(O)=CC(O)=C2C=C(O)C=1C1=CC=C(O)C(O)=C1 VEVZSMAEJFVWIL-UHFFFAOYSA-O 0.000 description 14
- 239000002904 solvent Substances 0.000 description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 9
- 241000196324 Embryophyta Species 0.000 description 8
- 235000007336 cyanidin Nutrition 0.000 description 7
- 238000001179 sorption measurement Methods 0.000 description 7
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 6
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 6
- 238000010828 elution Methods 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 238000009826 distribution Methods 0.000 description 5
- 238000004128 high performance liquid chromatography Methods 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 4
- 238000011109 contamination Methods 0.000 description 4
- 238000004811 liquid chromatography Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- 235000011301 Brassica oleracea var capitata Nutrition 0.000 description 3
- 244000178937 Brassica oleracea var. capitata Species 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 238000004587 chromatography analysis Methods 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 3
- 239000002244 precipitate Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 150000007513 acids Chemical class 0.000 description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000003795 desorption Methods 0.000 description 2
- -1 etc. is used Substances 0.000 description 2
- 229910052500 inorganic mineral Inorganic materials 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000011707 mineral Substances 0.000 description 2
- 125000003132 pyranosyl group Chemical group 0.000 description 2
- 235000011331 Brassica Nutrition 0.000 description 1
- 241000219198 Brassica Species 0.000 description 1
- RDFLLVCQYHQOBU-GPGGJFNDSA-O Cyanin Natural products O([C@H]1[C@H](O)[C@H](O)[C@H](O)[C@H](CO)O1)c1c(-c2cc(O)c(O)cc2)[o+]c2c(c(O[C@H]3[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O3)cc(O)c2)c1 RDFLLVCQYHQOBU-GPGGJFNDSA-O 0.000 description 1
- 125000005396 acrylic acid ester group Chemical group 0.000 description 1
- 229920006222 acrylic ester polymer Polymers 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- RDFLLVCQYHQOBU-ZOTFFYTFSA-O cyanin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC(C(=[O+]C1=CC(O)=C2)C=3C=C(O)C(O)=CC=3)=CC1=C2O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 RDFLLVCQYHQOBU-ZOTFFYTFSA-O 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 235000021110 pickles Nutrition 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
Landscapes
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
この発明は、酸性アントシアニン色素液に含まれる酸の
除去方法に関する。DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a method for removing acid contained in an acidic anthocyanin pigment liquid.
（従来の技術）
アントシアニンの分析方法は従来、いくつが知られてい
る［井高英−１特開昭６１−８５４７７号公報、ジャー
ナル・オブ・クロマトグラフィー、１５５巻、 ３８９
頁 （Ｍ、Ｗｉｌｌｉａｍｓ ａｔ ａｌ、、Ｊ、
ＣｈｒＯ謹ａｔｏｇｒ、、 １５５３８９ （１９７８
））、ジャーナル・オブ・クロマトグラフィー、１３２
巻、３４９頁（Ｍ。(Prior art) Several methods for analyzing anthocyanins are known [Hide Idaka-1 Japanese Patent Application Laid-open No. 85477/1985, Journal of Chromatography, Vol. 155, 389
Page (M. Williams at al., J.
ChrOsatogr,, 155389 (1978
)), Journal of Chromatography, 132
Volume, 349 pages (M.
１１１１ｋｉｎｓｏｎ ｅｔ ａｌ、、 Ｊ、Ｃｈｒｏ
ｍａｔｏｇｒ、、 １３２３４９ （１９７〕））、ツ
ァイトウンクス・ナチュールフォルシュンク、 ３５Ｃ
巻、 １６頁 （Ｎ、＾ｋａｖｉａ、Ｄ、５ｔｒａｃ
ｋ。1111kinson et al., J. Chro.
matogr,, 132349 (197])), Zeitounks Natureforschung, 35C
Volume, 16 pages (N, ^kavia, D, 5trac
k.
Ｚ、Ｎａｔｕｒｆｏｒｓｃｈ、、 ３５Ｃ１６（１９８
０）） ］ｅ Ｌかし、植物から抽出した複雑な混合
物からのアントシアニンの単離に関しては、井高英−１
特開昭６１−８５４７７号公報記載以外、全く知られて
いない［Ａ８ ヘンジエンら編、生化学における高速
液体クロマトグラフィー、６０１頁（Ｖ ＣＨ，西ドイ
ツ、１９８５）（原書名： Ａ、Ｈａｎｓｃｈａｎ、
Ｋ−Ｐ、Ｈｕｐｅ、 Ｆ。Z, Naturforsch, 35C16 (198
0))] e L. For the isolation of anthocyanins from complex mixtures extracted from plants, Hide Idaka-1
Nothing is known other than what is described in JP-A-61-85477 [A8 Hengeen et al., eds., High Performance Liquid Chromatography in Biochemistry, p. 601 (V CH, West Germany, 1985) (original title: A, Hanschan,
K-P, Hupe, F.
Ｌｏｔｔｓｐｅｉｃｈ、 Ｖ、Ｖｏｅｌｌｒ ａｄ、
Ｈｉｇｈ Ｐｓｒｆｏｒｍａｎｃｅ Ｌｉｑｕｉｄ Ｃ
ｈｒｏｍａｔｏｇｒａｐｈｙ ｉｎ Ｂｉｏｃｈｅｍｉ
ｓｔｒｙ、 ６０１ （１９８５）ＶＣＨ，ＦＲＧ）］
。Lottspeich, V. Voellr ad.
High Psrformance Liquid C
chromatography in biochemi
stry, 601 (1985) VCH, FRG)]
.
（発明が解決しようとする問題点〉
植物からの抽出液を特開昭６１−８５４７７号広報に記
載の方法に従って、高速液体クロマトグラフィー（ＨＰ
ＬＣ）を用いて分析した場合、分離しようとするアント
シアニンの近傍に他のアントシアニンが存在する場合に
は溶離液に強酸を使用する必要がある。その場合、溶出
液をそのまま乾固してアントシアニンを取り出すことは
出来ない。(Problems to be Solved by the Invention) Extracts from plants were subjected to high performance liquid chromatography (HP liquid chromatography) according to the method described in JP-A-61-85477.
In the case of analysis using LC), if other anthocyanins are present in the vicinity of the anthocyanin to be separated, it is necessary to use a strong acid as the eluent. In that case, it is not possible to directly dry the eluate and extract the anthocyanin.
本発明は上記の点に鑑み開発されたもので、酸性色素液
中から酸とアントシアニンとを分離し、分解を伴わずに
色素を純粋に取り出すことを目的としている。The present invention was developed in view of the above points, and its purpose is to separate acid and anthocyanin from an acidic dye solution and extract the dye purely without decomposition.
（問題点を解決するための手段）
上記目的達成の為、本発明は、酸性アントシアニン色素
液を樹脂吸着剤［例えば、アンバーライトＸＡＤ−２、
４、７、８、ＸＥ−２８４、ＭＣ■ゲルＣＨＰ２０Ｐ等
］を充填したカラムにマウントし、十分な量の水溶液［
例えば、０．５％トリフロロ酢酸（ＴＦＡ）、 １％酢
酸、水、 １％メタノール、３％クエン酸等、マウント
した色素がカラムから多量に溶離して来ない溶離液］で
洗浄し、当初含まれていた酸を除いた後、色素が溶離し
うる溶離液を用いて色素を溶出する（ここで、当初含ま
れていた酸が例えば、リン酸、塩酸等の鉱酸類であ）て
、溶離液に酸を含まない溶媒を使用すれば脱酸を行った
ことになり、一方、ＴＦＡ、・酢酸等を含む溶媒を使用
すれば酸交換を行゛つたことになる。）。ここで帰られ
た溶離液中には多量の樹脂成分が溶出してきており、こ
のまま、溶媒を乾固した場合、残漬として色素と共に樹
脂成分が残ってくる。この残漬を、例えば、１％塩ｅｊ
！／メタノールに溶解しエーテルを加えて沈澱或は、結
晶として得ようとしても共存する樹脂成分が混入してき
て、アントシアニンを純粋に取り出すことは出来ない。(Means for Solving the Problems) In order to achieve the above object, the present invention provides an acidic anthocyanin pigment solution to a resin adsorbent [for example, Amberlite XAD-2,
4, 7, 8,
For example, wash with an eluent that does not elute a large amount of the mounted dye from the column, such as 0.5% trifluoroacetic acid (TFA), 1% acetic acid, water, 1% methanol, 3% citric acid, etc. After removing the acid contained in the dye, the dye is eluted using an eluent that can elute the dye (here, the acid originally contained is, for example, a mineral acid such as phosphoric acid or hydrochloric acid). If a solvent containing no acid is used in the liquid, deacidification has been performed, whereas if a solvent containing TFA, acetic acid, etc. is used, acid exchange has been performed. ). A large amount of resin components have eluted into the eluent returned here, and if the solvent is dried to solidify as it is, the resin components will remain as a residue along with the dye. For example, add 1% salt to this residue.
! Even if an attempt is made to obtain the anthocyanin as a precipitate or crystal by dissolving it in methanol and adding ether, the coexisting resin components will be mixed in, making it impossible to extract the anthocyanin in a pure form.
従って、次に、逆相吸着分配型カラム［ＣＩｍ （○Ｄ
Ｓ）、Ｃｍ、ＥＴＨ等］を用いた液体クロマトグラフィ
ー（ＬＣ）による樹脂吸着剤混入成分を除去することが
必要である。即ち、色素溶離液を逆相吸着分配型カラム
にマウントする・ この場合、色素溶離液の極性によっ
てはマウント中に色素が移動する可能性があるので、少
し濃縮するか或は、水を加えて極性を上げる必要がある
場合がある。Therefore, next, a reversed-phase adsorption/distribution column [CIm (○D
It is necessary to remove components mixed with the resin adsorbent by liquid chromatography (LC) using [S), Cm, ETH, etc.]. That is, the dye eluent is mounted on a reversed-phase adsorption/distribution column.In this case, depending on the polarity of the dye eluent, the dye may migrate during mounting, so it may be necessary to concentrate it slightly or add water. It may be necessary to increase the polarity.
マウント後、色素が移動して流出に至らない程度の溶ａ
（例えば、水、０．５％ＴＦＡ、２％アセトニトリル、
５％エタノール、５％プロピレングリコール等）で洗浄
し、樹脂吸着剤混入成分を除去した後、溶離液で色素を
溶出する。ここで、色素を個体（沈澱又は、結晶）とし
て得たい場合には、溶離液に酸を用いないか或は、揮発
性（ＴＦＡ、酢酸等）の酸を用いる０色素を濃厚液とし
て得たい場合には不揮発性（クエン酸、鉱酸等）の酸を
用いても良い。After mounting, lyse to an extent that the dye does not move and flow out.
(For example, water, 0.5% TFA, 2% acetonitrile,
After washing with 5% ethanol, 5% propylene glycol, etc.) to remove components mixed with the resin adsorbent, the dye is eluted with an eluent. If you want to obtain the dye as a solid (precipitate or crystal), either do not use an acid as the eluent or use a volatile acid (TFA, acetic acid, etc.) to obtain the dye as a concentrated liquid. In some cases, non-volatile acids (citric acid, mineral acids, etc.) may be used.
この一連の操作によって、酸性アントシアニン色素液か
ら、これに当初含まれる酸を除去する事を特徴とするア
ントシアニン色素のｔａ製製法法ある。There is a TA manufacturing method for anthocyanin pigments, which is characterized by removing the acid initially contained in the acidic anthocyanin pigment liquid through this series of operations.
（発明の効果）
以上述べたように、この発明は酸性アントシアニン色素
液から、２段階カラム説着法Ｃ゛よって酸の除去或は、
交換を可能とした。このことによって、植物から抽出し
た！！謡なアン！・シアニン混合物から、アントシアニ
ンを各々単離することが可能となった。(Effects of the Invention) As described above, this invention removes acid from an acidic anthocyanin pigment solution by a two-step column deposition method C.
made exchange possible. By this, extracted from plants! ! Singing Anne! - It became possible to isolate each anthocyanin from a cyanin mixture.
又、従来、良品着色用アントシアニン色素は、植物から
溶媒抽出したままの粗色素を、植物成分とアントシアニ
ン色素混合物とを荒分けする目的で吸着樹脂カラムによ
る脱着によって粗精製を行い、濃縮色素ジュース（１５
％プロピレングリコール、 １％クエン酸含有）として
市販されているが、上記した様に吸着樹脂カラムによる
脱着によって大量の樹脂成分が市販の濃縮色素ジュース
中に含まれていることが明かとなった。このことは樹脂
成分の毒性の有無を含めて人間の健康に関する重要な問
題を提起するものである。即ち、樹脂成分の除去の為、
新たな色素の製法を検討する必要がある。その場合、本
方法に則って逆相吸着分配型カラムによる樹脂成分の除
去を試みるか或は、植物から溶媒抽出したままの粗色素
を、植物成分とアントシアニン色素混合物とを荒分けす
る目的で、直接、逆相吸着分配型カラムによる粗精製を
行い、濃縮色素ジュースとすることによって、樹脂成分
の混入を防ぐことが可能となる（この場合、逆相吸着分
配型充填剤による成分の溶出はない事が５００ＭＨｚ
’ＨＮＭＲによって確認した）。Conventionally, good quality anthocyanin pigments for coloring have been obtained by crudely refining the crude pigments extracted from plants with solvents by desorption using an adsorption resin column in order to roughly separate the plant components and the anthocyanin pigment mixture, and then producing concentrated pigment juice ( 15
% propylene glycol, 1% citric acid), but as mentioned above, it has become clear that a large amount of resin components are contained in the commercially available concentrated pigment juice due to desorption using an adsorption resin column. This raises important questions regarding human health, including the toxicity of resin components. That is, in order to remove the resin component,
It is necessary to consider new methods for producing pigments. In that case, try to remove the resin component using a reversed-phase adsorption/distribution column according to this method, or roughly separate the crude pigment extracted from the plant with a solvent into the plant component and the anthocyanin pigment mixture. By directly performing rough purification using a reversed-phase adsorption/distribution column to create a concentrated pigment juice, it is possible to prevent the contamination of resin components (in this case, there is no elution of components by the reversed-phase adsorption/distribution type packing material). thing is 500MHz
' Confirmed by HNMR).
次に、この発明の酸性アントシアニン色素液に含まれる
酸の除去方法を実施例によって更に詳細に説明する。Next, the method for removing acid contained in the acidic anthocyanin pigment liquid of the present invention will be explained in more detail with reference to Examples.
実施例１
赤キャベツ（学名： Ｂｒａｓｓｉｃａ ｏｌｅｒａ
ｃｅａ Ｌ、 ｖａｒ、 ｒｕｂｒａ）には２０種類以
上のアントシアニンが含まれている事がＨＰＬＣ分析の
結果間らかになったが、その中の３１！１１７１のアン
トシアニンをＨＰＬＣで分取し、その構造決定を行った
。すなわち、３−０− ［６−０−ｐ−クマリル−２−
０−（２−〇−シナビルーβ−Ｄ−グルコピラノシル）
−β−Ｄ−グルコピラノシル］−５−０−（β−Ｄ−グ
ルコピラノシル）シアニジン（Ｉ ） ［３−０−［
６−Ｏ−ｐ−ｃｏｕｍ６−０−ｐ−ｃｏｕ （２−０−
ｓｉｎａｐｙｌ−β−トｇｌｕｃｏｐｙｒａｎｏｓｙｌ
）−β−Ｄ−ｇｌｕｃｏｐｙｒａｎｏｓｙｌｌ−５−０
−（β−Ｄ−ｇｌｕｃ。Example 1 Red cabbage (scientific name: Brassica olera
HPLC analysis revealed that cea L, var, rubra) contain more than 20 types of anthocyanins, and among them, 31!1171 anthocyanins were fractionated by HPLC and their structures were determined. made the decision. That is, 3-0-[6-0-p-coumaryl-2-
0-(2-〇-cinnabi-β-D-glucopyranosyl)
-β-D-glucopyranosyl]-5-0-(β-D-glucopyranosyl)cyanidin (I) [3-0-[
6-O-p-coum6-0-p-cou (2-0-
sinapyl-β-glucopyranosyl
)-β-D-glucopyranosyl-5-0
-(β-D-gluc.
ｐｙｒａｎｏｓｙｌ）ｃｙａｎｉｄｉｎｌ、３−０−
［６−０−フエルリル−２−０−（２−０−シナビル−
β−Ｄ−グルコピラノシル）−β−Ｄ−グルコピラノシ
ル］−５−０−（β−Ｄ−グルコピラノシル）シアニジ
ン（Ｉ Ｉ）及び、 ［３−０−［６−Ｏ−ｆｅｒｕｌ
ｙｌ−２−０−（２−０−ｓｉｎａｐｙｌ−β−Ｄ−ｇ
ｌｕｃｏｐｙｒａｎｏｓｙｌ）−β−１］−ｇｌｕｃＯ
ＰＹｒａｎｏｓｙｌｌ−５−０−（β−Ｄ−ｇｌｕｃａ
ｐｙｒａｎｏｓｙｌ）ｃｙａｎｉｄｉｎｌ、３−０−
［６−０−シナビル−２−０−（２−０−シナビル−β
−Ｄ−グルコピラノシル）−β−Ｄ−グルコピラノシル
］−５−０−（β−Ｄ−グルコピラノシル）シアニジン
（Ｉ Ｉ Ｉ ） ［３−０−［６−Ｏ−ｓ １ｎａ
ｐｙｌ−２−０−（２−０−ｓ １ｎａｐｙｌ−β−Ｄ
−ｇｌｕｃｏｐｙｒａｎ。pyranosyl) cyanidinl, 3-0-
[6-0-Feruryl-2-0-(2-0-Cinavir-
β-D-glucopyranosyl)-β-D-glucopyranosyl]-5-0-(β-D-glucopyranosyl)cyanidin (II) and [3-0-[6-O-ferul
yl-2-0-(2-0-sinapyl-β-D-g
lucopyranosyl)-β-1]-glucO
PYranosyl-5-0-(β-D-gluca
pyranosyl) cyanidinl, 3-0-
[6-0-Cinavir-2-0-(2-0-Cinavir-β
-D-glucopyranosyl)-β-D-glucopyranosyl]-5-0-(β-D-glucopyranosyl)cyanidin (I II I) [3-0-[6-O-s 1na
pyl-2-0-(2-0-s 1napyl-β-D
-glucopyran.
５ｙｌ）−β−Ｄ−ｇｌｕｃｏｐｙｒａｎｏａｙｌｌ−
５−０−（β−ｏ−ｇｔｕｃｏｐｙｒａｎｏｓｙｌ）ｃ
ｙａｎｉｄｉｎｌを特開詔６１−８５４７７号公報記載
の方法によってＨＰＬＣを用いて分取した。5yl)-β-D-glucopyranoayll-
5-0-(β-o-gtucopyranosyl)c
Yanidinl was fractionated using HPLC according to the method described in Japanese Patent Application Laid-open No. 61-85477.
溶離液の酸成分として１．５％リン酸を使用した。1.5% phosphoric acid was used as the acid component of the eluent.
この溶出液をエバポレータで低温減圧乾固を行った場合
、 （Ｉ）〜（ＩＩＩ）は濃縮された酸で分解してしま
う。そこで、アンバーライトＸＡＤ−７カラムに各々の
色素を含む溶離液をマウントし、充分な量の０．５％Ｔ
ＦＡで洗浄した後、３０％メタノール水溶液（０，５％
ＴＦＡを含む）で色素を溶出した。約３０％減圧濃縮し
てメタノールを除いたｉｋ ＯＤ ｓカラムにマウント
し０．５％ＴＦＡで充分洗浄して樹脂溶出成分を除いた
後、１０％メタノールで色素を溶出した。得られた溶出
液を低温減圧乾固して各々の色素をＴＦＡ塩として得た
。これを更に１％塩酸／メタノールに溶かし、エーテル
を加えて色素を塩化物として沈澱させた。この沈澱を濾
過、乾燥して純粋なアントシアニンを塩化物として得た
。When this eluate is dried under low temperature and reduced pressure in an evaporator, (I) to (III) are decomposed by concentrated acid. Therefore, the eluent containing each dye was mounted on an Amberlite XAD-7 column, and a sufficient amount of 0.5% T was added.
After washing with FA, 30% methanol aqueous solution (0.5%
The dye was eluted with TFA (containing TFA). The column was mounted on an ik OD s column that had been concentrated under reduced pressure by about 30% to remove methanol, and washed thoroughly with 0.5% TFA to remove resin-eluted components, and then the dye was eluted with 10% methanol. The obtained eluate was dried under reduced pressure at low temperature to obtain each dye as a TFA salt. This was further dissolved in 1% hydrochloric acid/methanol and ether was added to precipitate the dye as a chloride. The precipitate was filtered and dried to obtain pure anthocyanin as chloride.
実施例２
３−０− ［６−０−シｆビル−２−０−（ｐ’−Ｄ−
グルコピラノシル）−β−Ｄ−グルコピラノシル］−５
−０−（β−Ｄ−グルコピラノシル）シアニジ＞ （Ｉ
Ｖ ） ｒ ３−０−［６−Ｏ−ｓｉｎａｐＨ−２
−［１−（β−Ｄ−ｇｌｕｃｏｐｙｒａｎｏｓｙｌ）−
β−Ｄ−ｇｌｕｃｏｐｙｒａｎｏｓｙｌｌ−５−０−（
β−Ｄ−ｇｌｕｃｏｐｙｒａｎｏｓｙｌ）ｃｙａｎｉｄ
ｉｎｌ （塩化物）５ｍｇをＸＡＤ−７カラムにマウン
ト、洗浄、溶出後、溶媒を減圧濃縮し、色素を残漬とし
て回収したく収量６５ｍｇ）、 この残漬の５００Ｍ
Ｈｚ ’ＨＮＭＲスペクトルを第１図に示す。０．７
〜１．８Ｐ ｐｍ ＋： Ｘ Ａ Ｄ −７樹脂（アク
リル酸エステル・ポリマー）に由来するピークが１ＩＩ
Ｈされる。この１１ｔＭｔｔｏ、 ５％ＴＦＡに溶解
しＯＤＳカラムにマウント後、０．５％ＴＦＡで洗浄、
１０％エタノール（０，５％ＴＦＡを含む）で溶出後
、溶媒を減圧濃縮し、色素を残漬として回収した（収量
５゜０ｍｇ）。この残漬の５００ＭＨｚ ’ＨＮＭＲ
スペクトルを第２図に示す、０．７〜１．８ｐｐｍにＸ
ＡＤ−７樹脂（アクリル餞エステル・ポリマー）に由来
するピークは消失しており純粋な色素がほぼ定量的に回
収された。この色素を１％塩酸／メタノールに溶解し、
エーテルで色素を沈澱させ、濾別、乾燥後、純粋な色素
を塩化物として得た（収ｆｉ４．２ｍｇ、収率８４％）
。Example 2 3-0-[6-0-cyf-2-0-(p'-D-
glucopyranosyl)-β-D-glucopyranosyl]-5
-0-(β-D-glucopyranosyl)cyanidi> (I
V) r3-0-[6-O-sinapH-2
-[1-(β-D-glucopyranosyl)-
β-D-glucopyranosyl-5-0-(
β-D-glucopyranosyl)cyanid
After mounting 5 mg of inl (chloride) on an XAD-7 column, washing, and elution, the solvent was concentrated under reduced pressure to recover the dye as a residue (yield: 65 mg).
The Hz'HNMR spectrum is shown in FIG. 0.7
~1.8P pm +: The peak derived from XAD-7 resin (acrylic acid ester polymer) is 1II
He gets fucked. This 11tMtto was dissolved in 5% TFA, mounted on an ODS column, washed with 0.5% TFA,
After elution with 10% ethanol (containing 0.5% TFA), the solvent was concentrated under reduced pressure and the dye was recovered as a residue (yield: 5.0 mg). 500MHz 'HNMR of this residual
The spectrum is shown in Figure 2,
The peak derived from AD-7 resin (acrylic ester polymer) disappeared, and pure dye was recovered almost quantitatively. Dissolve this dye in 1% hydrochloric acid/methanol,
After precipitating the dye with ether, filtering and drying, the pure dye was obtained as a chloride (fi 4.2 mg, yield 84%).
.
実施例３
赤キャベツから抽出したアントシアニン粗色素（乾燥重
量１ｇ）をＭＣＩゲルＣＨＰ２０Ｐにマウント後、洗浄
、溶出した後、低温減圧濃縮した結果、乾燥重量０．７
７ｇを得た。５２０ｎｍにおける吸光度によって色素の
回収率を測定したところ８０％の回収率であった。、こ
こで回収した色素をＯＤＳカラムにマウント後、洗浄、
溶出し、低温減圧乾固した結果、０．３ｇの粗色素を得
た。Example 3 Crude anthocyanin pigment (dry weight 1 g) extracted from red cabbage was mounted on MCI gel CHP20P, washed, eluted, and concentrated under reduced pressure at low temperature, resulting in a dry weight of 0.7
7g was obtained. The recovery rate of the dye was measured by absorbance at 520 nm, and the recovery rate was 80%. , After mounting the recovered dye on an ODS column, washing,
As a result of elution and drying under reduced pressure at low temperature, 0.3 g of crude pigment was obtained.
吸光度による色素の回収率は８７％であった。The dye recovery rate based on absorbance was 87%.
一方、赤キャベツから抽出したアントシアニン粗色素（
乾燥重量１ｇ）をＯＤＳカラムにマウント後、洗浄、溶
出し、低温減圧乾固した結果、０゜４８ｇの粗色素を得
た。吸光度による色素の回収率は８２％であった。On the other hand, anthocyanin crude pigment extracted from red cabbage (
After mounting the dye (dry weight: 1 g) on an ODS column, washing, elution, and drying under reduced pressure at low temperature, 0.48 g of crude dye was obtained. The dye recovery rate based on absorbance was 82%.
この結果から、ＭＣＩゲルＣＨＰ２０Ｐによる処理はか
なりのゲル成分の混入をもたらしている。From this result, treatment with MCI gel CHP20P resulted in considerable contamination of gel components.
このゲル成分の混入はＯＤＳカラム処理によってかなり
除去されている。色素以外の植物成分の混入は、ゲル処
理のみ、ＯＤＳカラム処理のみ、ゲル・０Ｄ８２段処理
の順に減少している。This gel component contamination was largely removed by ODS column treatment. The contamination of plant components other than pigments decreased in the order of gel treatment only, ODS column treatment only, and gel/0D8 two-stage treatment.
第１図は３−０− ［６−０−シナビル−２−０−（β
−Ｄ−グルコピラノシル）−β−Ｄ−グルコピラノシル
］−５−０−（β−Ｄ−グルコピラノシル）シアニジン
（Ｉ Ｖ ） ［３−０−［６−Ｏ−ｓｉｎａｐｙｌ
−２−０−（β−Ｄ−ｇｌｕｃｏｐｙｒａｎｏｓｙｌ）
−β−Ｄ−ｇｌｕｃｏｐｙｒａｎｏｓｙ１１−５−０−
（β−Ｄ−ｇｌｕｃｏｐｙｒａｎｏｓｙｌ） ｃｙａｎ
ｉｄｉｎ ］をＸＡＤ−７カラムにマウント、洗浄、溶
出後、溶媒を減圧濃縮し、色素を残漬として回収した。
この残漬の５００ＭＨｚ ’ＨＮＭＲスペクトルであ
る。
第２図は３−０− ［６−０−シナビル−２−〇−（β
−Ｄ−グルコピラノシル）−β−Ｄ−グルコピラノシル
］ −５−０−（β−Ｄ−グルコピラノシル）シアニジ
ン（Ｉ Ｖ ） ［３−０−［６−Ｏ−ｓｉｎａｐｙ
ｌ−２−０−（β−Ｄ−ｇｌｕｃｏｐｙｒａｎｏｓｙｌ
）−β−Ｄ−ｇｌｕｃｏｐｙｒａｎｏｓｙ１１−５−０
−（β−Ｄ−ｇｌｕｃｏｐｙｒａｎｏｓｙｌ） ｃｙａ
ｎｉｄｉｎ ］をＸＡＤ−７カラムにマウント、洗浄、
溶出後、溶媒を減圧濃縮し、色素を残漬として回収した
。この残漬を０． ５％ＴＦＡに溶解しＯＤＳカラムに
マウント後、０．５％ＴＦＡで洗浄、 １０％エタノー
ル（０，５％ＴＦＡを含む）で溶出後、溶媒を減圧濃縮
し、色素を残漬として回収した。この残漬の５００ＭＨ
ｚ ’ＨＮＭＲスペクトルである。Figure 1 shows 3-0-[6-0-cinavir-2-0-(β
-D-glucopyranosyl)-β-D-glucopyranosyl]-5-0-(β-D-glucopyranosyl)cyanidin (IV) [3-0-[6-O-sinapyl
-2-0-(β-D-glucopyranosyl)
-β-D-glucopyranosy11-5-0-
(β-D-glucopyranosyl) cyan
idin] was mounted on an XAD-7 column, washed, and eluted, the solvent was concentrated under reduced pressure and the dye was recovered as a residual dye. This is a 500 MHz 'HNMR spectrum of this residual material. Figure 2 shows 3-0-[6-0-cinavir-2-〇-(β
-D-glucopyranosyl)-β-D-glucopyranosyl] -5-0-(β-D-glucopyranosyl)cyanidin (IV) [3-0-[6-O-sinapy
l-2-0-(β-D-glucopyranosyl
)-β-D-glucopyranosy11-5-0
-(β-D-glucopyranosyl) cya
Nidin] was mounted on an XAD-7 column, washed,
After elution, the solvent was concentrated under reduced pressure and the dye was recovered as a residue. This residual pickle is 0. The dye was dissolved in 5% TFA and mounted on an ODS column, washed with 0.5% TFA, and eluted with 10% ethanol (containing 0.5% TFA). The solvent was concentrated under reduced pressure and the dye was recovered as a residual dye. 500MH of this residual
z'HNMR spectrum.
Claims (1)
ムにマウントし、水溶液で洗浄することによって脱酸し
た後、溶離液で色素を溶出し、得られた溶出液を続いて
、逆相吸着分配型カラムにマウントした後、水溶液で洗
浄することによって混入した樹脂吸着剤を除いた後、溶
離液で色素を溶出し、その溶出液を濃縮する事によって
アントシアニン色素を得る方法The acidic anthocyanin dye solution is mounted on a column packed with a resin adsorbent, deoxidized by washing with an aqueous solution, and then the dye is eluted with an eluent. A method of obtaining anthocyanin pigments by mounting them on a substrate, removing the mixed resin adsorbent by washing with an aqueous solution, eluating the pigments with an eluent, and concentrating the eluate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62113201A JPS63278971A (en) | 1987-05-09 | 1987-05-09 | Removal of acid from acidic anthocyanin pigment solution |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62113201A JPS63278971A (en) | 1987-05-09 | 1987-05-09 | Removal of acid from acidic anthocyanin pigment solution |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS63278971A true JPS63278971A (en) | 1988-11-16 |
Family
ID=14606116
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP62113201A Pending JPS63278971A (en) | 1987-05-09 | 1987-05-09 | Removal of acid from acidic anthocyanin pigment solution |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63278971A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005510512A (en) * | 2001-11-09 | 2005-04-21 | メドパレット ファーマシューティカルズ エイエス | Method for producing anthocyanin-containing product |
| JP2009298926A (en) * | 2008-06-13 | 2009-12-24 | Tochigi Prefecture | Method for purifying anthocyanin pigment from preliminary pickling liquid of eggplant |
-
1987
- 1987-05-09 JP JP62113201A patent/JPS63278971A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005510512A (en) * | 2001-11-09 | 2005-04-21 | メドパレット ファーマシューティカルズ エイエス | Method for producing anthocyanin-containing product |
| JP2009298926A (en) * | 2008-06-13 | 2009-12-24 | Tochigi Prefecture | Method for purifying anthocyanin pigment from preliminary pickling liquid of eggplant |
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