CN101560155A - Method for purifying cynarin in artichoke - Google Patents
Method for purifying cynarin in artichoke Download PDFInfo
- Publication number
- CN101560155A CN101560155A CNA200910107323XA CN200910107323A CN101560155A CN 101560155 A CN101560155 A CN 101560155A CN A200910107323X A CNA200910107323X A CN A200910107323XA CN 200910107323 A CN200910107323 A CN 200910107323A CN 101560155 A CN101560155 A CN 101560155A
- Authority
- CN
- China
- Prior art keywords
- arithoke
- purification
- obtains
- cynarin
- liquid
- 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
- 238000000034 method Methods 0.000 title claims abstract description 44
- 235000019106 Cynara scolymus Nutrition 0.000 title claims abstract description 15
- 244000019459 Cynara cardunculus Species 0.000 title claims abstract description 14
- 229950009125 cynarine Drugs 0.000 title abstract description 12
- YDDUMTOHNYZQPO-UHFFFAOYSA-N 1,3-bis{[(2E)-3-(3,4-dihydroxyphenyl)prop-2-enoyl]oxy}-4,5-dihydroxycyclohexanecarboxylic acid Natural products OC1C(O)CC(C(O)=O)(OC(=O)C=CC=2C=C(O)C(O)=CC=2)CC1OC(=O)C=CC1=CC=C(O)C(O)=C1 YDDUMTOHNYZQPO-UHFFFAOYSA-N 0.000 title abstract description 9
- YDDUMTOHNYZQPO-RVXRWRFUSA-N Cynarine Chemical compound O([C@@H]1C[C@@](C[C@H]([C@@H]1O)O)(OC(=O)\C=C\C=1C=C(O)C(O)=CC=1)C(O)=O)C(=O)\C=C\C1=CC=C(O)C(O)=C1 YDDUMTOHNYZQPO-RVXRWRFUSA-N 0.000 title abstract description 9
- YDDUMTOHNYZQPO-BKUKFAEQSA-N cynarine Natural products O[C@H]1C[C@@](C[C@H](OC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)(OC(=O)C=Cc3ccc(O)c(O)c3)C(=O)O YDDUMTOHNYZQPO-BKUKFAEQSA-N 0.000 title abstract description 9
- SITQVDJAXQSXSA-CEZRHVESSA-N Cynarin Natural products O[C@@H]1C[C@@](C[C@H](O)[C@H]1OC(=O)C=Cc2ccc(O)c(O)c2)(OC(=O)C=Cc3cccc(O)c3O)C(=O)O SITQVDJAXQSXSA-CEZRHVESSA-N 0.000 title abstract description 8
- 235000016520 artichoke thistle Nutrition 0.000 title abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 32
- 239000013078 crystal Substances 0.000 claims abstract description 29
- 239000000047 product Substances 0.000 claims abstract description 22
- 238000001914 filtration Methods 0.000 claims abstract description 20
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 16
- 238000001471 micro-filtration Methods 0.000 claims abstract description 13
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 12
- 238000001035 drying Methods 0.000 claims abstract description 8
- 239000011347 resin Substances 0.000 claims abstract description 7
- 229920005989 resin Polymers 0.000 claims abstract description 7
- 239000000706 filtrate Substances 0.000 claims abstract description 6
- 238000005406 washing Methods 0.000 claims abstract 2
- 239000007788 liquid Substances 0.000 claims description 45
- 238000000746 purification Methods 0.000 claims description 29
- 238000004587 chromatography analysis Methods 0.000 claims description 20
- 238000010828 elution Methods 0.000 claims description 17
- 239000012528 membrane Substances 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 16
- 229910052500 inorganic mineral Inorganic materials 0.000 claims description 14
- 239000011707 mineral Substances 0.000 claims description 14
- 238000005119 centrifugation Methods 0.000 claims description 11
- 238000000151 deposition Methods 0.000 claims description 11
- 239000003960 organic solvent Substances 0.000 claims description 10
- 238000001953 recrystallisation Methods 0.000 claims description 9
- 239000012141 concentrate Substances 0.000 claims description 8
- 238000000502 dialysis Methods 0.000 claims description 8
- 150000008442 polyphenolic compounds Chemical class 0.000 claims description 8
- 235000013824 polyphenols Nutrition 0.000 claims description 8
- 239000002253 acid Substances 0.000 claims description 7
- 238000002425 crystallisation Methods 0.000 claims description 6
- 230000008025 crystallization Effects 0.000 claims description 6
- 239000012535 impurity Substances 0.000 claims description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 5
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 4
- 238000013375 chromatographic separation Methods 0.000 claims description 4
- 239000001117 sulphuric acid Substances 0.000 claims description 4
- 235000011149 sulphuric acid Nutrition 0.000 claims description 4
- 239000013049 sediment Substances 0.000 claims description 3
- 238000000638 solvent extraction Methods 0.000 claims description 2
- 238000001179 sorption measurement Methods 0.000 claims description 2
- 239000000284 extract Substances 0.000 abstract description 7
- 239000012043 crude product Substances 0.000 abstract 2
- 238000010521 absorption reaction Methods 0.000 abstract 1
- 238000009776 industrial production Methods 0.000 abstract 1
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 6
- 238000000605 extraction Methods 0.000 description 6
- 239000012071 phase Substances 0.000 description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 5
- 239000001913 cellulose Substances 0.000 description 5
- 229920002678 cellulose Polymers 0.000 description 5
- 238000004128 high performance liquid chromatography Methods 0.000 description 5
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 4
- 239000003463 adsorbent Substances 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 235000006708 antioxidants Nutrition 0.000 description 4
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 4
- 239000000945 filler Substances 0.000 description 4
- 210000004185 liver Anatomy 0.000 description 4
- 239000001814 pectin Substances 0.000 description 4
- 229920001277 pectin Polymers 0.000 description 4
- 235000010987 pectin Nutrition 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 239000001648 tannin Substances 0.000 description 4
- 235000018553 tannin Nutrition 0.000 description 4
- 229920001864 tannin Polymers 0.000 description 4
- 239000003643 water by type Substances 0.000 description 4
- CWVRJTMFETXNAD-FWCWNIRPSA-N 3-O-Caffeoylquinic acid Natural products O[C@H]1[C@@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-FWCWNIRPSA-N 0.000 description 3
- PZIRUHCJZBGLDY-UHFFFAOYSA-N Caffeoylquinic acid Natural products CC(CCC(=O)C(C)C1C(=O)CC2C3CC(O)C4CC(O)CCC4(C)C3CCC12C)C(=O)O PZIRUHCJZBGLDY-UHFFFAOYSA-N 0.000 description 3
- 241000725303 Human immunodeficiency virus Species 0.000 description 3
- CWVRJTMFETXNAD-KLZCAUPSSA-N Neochlorogenin-saeure Natural products O[C@H]1C[C@@](O)(C[C@@H](OC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O CWVRJTMFETXNAD-KLZCAUPSSA-N 0.000 description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- CWVRJTMFETXNAD-JUHZACGLSA-N chlorogenic acid Chemical compound O[C@@H]1[C@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-JUHZACGLSA-N 0.000 description 3
- 229940074393 chlorogenic acid Drugs 0.000 description 3
- FFQSDFBBSXGVKF-KHSQJDLVSA-N chlorogenic acid Natural products O[C@@H]1C[C@](O)(C[C@@H](CC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O FFQSDFBBSXGVKF-KHSQJDLVSA-N 0.000 description 3
- 235000001368 chlorogenic acid Nutrition 0.000 description 3
- BMRSEYFENKXDIS-KLZCAUPSSA-N cis-3-O-p-coumaroylquinic acid Natural products O[C@H]1C[C@@](O)(C[C@@H](OC(=O)C=Cc2ccc(O)cc2)[C@@H]1O)C(=O)O BMRSEYFENKXDIS-KLZCAUPSSA-N 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- MGYBYJXAXUBTQF-FOBVWLSUSA-N 2-(3,4-dihydroxyphenyl)-5-hydroxy-7-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-[[(2r,3r,4r,5r,6s)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxymethyl]oxan-2-yl]oxychromen-4-one Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](OC=2C=C3C(C(C=C(O3)C=3C=C(O)C(O)=CC=3)=O)=C(O)C=2)O1 MGYBYJXAXUBTQF-FOBVWLSUSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- LOUPRKONTZGTKE-WZBLMQSHSA-N Quinine Chemical compound C([C@H]([C@H](C1)C=C)C2)C[N@@]1[C@@H]2[C@H](O)C1=CC=NC2=CC=C(OC)C=C21 LOUPRKONTZGTKE-WZBLMQSHSA-N 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 235000012000 cholesterol Nutrition 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- IVTMALDHFAHOGL-UHFFFAOYSA-N eriodictyol 7-O-rutinoside Natural products OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OC=2C=C3C(C(C(O)=C(O3)C=3C=C(O)C(O)=CC=3)=O)=C(O)C=2)O1 IVTMALDHFAHOGL-UHFFFAOYSA-N 0.000 description 2
- MGYBYJXAXUBTQF-FSFUDZCRSA-N luteolin 7-O-rutinoside Natural products C[C@@H]1O[C@H](OC[C@H]2O[C@@H](Oc3cc(O)c4C(=O)C=C(Oc4c3)c5ccc(O)c(O)c5)[C@H](O)[C@@H](O)[C@@H]2O)[C@@H](O)[C@H](O)[C@H]1O MGYBYJXAXUBTQF-FSFUDZCRSA-N 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000011034 membrane dialysis Methods 0.000 description 2
- MVTFTBIUYLXNKP-UHFFFAOYSA-N scolimoside Natural products COc1cc(O)c2C(=O)C=C(Oc2c1)c3ccc(OC4OC(COC5OC(C)C(O)C(O)C5O)C(O)C(O)C4O)c(O)c3 MVTFTBIUYLXNKP-UHFFFAOYSA-N 0.000 description 2
- NRHBUMVXUGSZAT-IVUGMLEJSA-N (3R,5R)-3-[(E)-3-(3,4-dihydroxyphenyl)prop-2-enoyl]-1,3,4,5-tetrahydroxycyclohexane-1-carboxylic acid Chemical class O[C@@H]1CC(O)(C[C@@](O)(C1O)C(=O)\C=C\c1ccc(O)c(O)c1)C(O)=O NRHBUMVXUGSZAT-IVUGMLEJSA-N 0.000 description 1
- YDDUMTOHNYZQPO-BBLPPJRLSA-N 1,3-di-O-caffeoylquinic acid Natural products O[C@@H]1C[C@@](C[C@@H](OC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)(OC(=O)C=Cc1ccc(O)c(O)c1)C(O)=O YDDUMTOHNYZQPO-BBLPPJRLSA-N 0.000 description 1
- SDYWXFYBZPNOFX-UHFFFAOYSA-N 3,4-dichloroaniline Chemical compound NC1=CC=C(Cl)C(Cl)=C1 SDYWXFYBZPNOFX-UHFFFAOYSA-N 0.000 description 1
- 241000050051 Chelone glabra Species 0.000 description 1
- 235000001258 Cinchona calisaya Nutrition 0.000 description 1
- 244000309023 Cynara scolymus Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000234435 Lilium Species 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 230000036436 anti-hiv Effects 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- LOUPRKONTZGTKE-UHFFFAOYSA-N cinchonine Natural products C1C(C(C2)C=C)CCN2C1C(O)C1=CC=NC2=CC=C(OC)C=C21 LOUPRKONTZGTKE-UHFFFAOYSA-N 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000003337 fertilizer Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 229960000948 quinine Drugs 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
Landscapes
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a method for purifying cynarin in an artichoke. The method comprises the following steps of: extracting the artichoke leaves by an ethanol or acetone solution, concentrating and filtering the extract by water, carrying out micro-filtration treatment to the filtrate, adjusting the pH value of a dialyzate to be less than 6, washing and separating the dialyzate by a macroporous absorption resin chromatographic column, eluting the chromatographic column by the ethanol solution with the volume percentage concentration of 10-18%, thus obtaining a polyphenol-contained eluant, eluting the chromatographic column by the ethanol solution with the volume percentage concentration of 50-70%, thus obtaining a cynarin-contained eluant, concentrating and crystallizing the cynarin-contained eluant to obtain a cynarin moist crystal, drying the moist crystal to obtain the cynarin crude product, and re-crystallizing and drying the cynarin moist crystal to obtain the cynarin product. The cynarin crude product obtained by the method has a purity of more than 20% and can be provided to the clients with corresponding requirements. The method has simple process and is suitable for industrial production.
Description
Technical field
The present invention relates to a kind of method of extracting purifying arithoke element, more particularly, relate to a kind of method of from leaf of Cynara scolymus L, extracting purifying arithoke element.
Background technology
Choke (Cynara scolymus L.) belongs to per nnial herb for the composite family cynara scolymus, and another name arithoke, cynara scolymus, French lily originate in the area between north African and the Mediterranean Sea the east, and plantation history in several thousand is arranged.Choke has very high nutritive value, is a kind of high-grade health-care vegetable, and its leaf contains arithoke element, chlorogenic acid etc. and has the active skull cap components that protects the liver, protects liver, reducing cholesterol effect.Leaf of Cynara scolymus L is taken as feed or fertilizer use mostly at present, and the value of its arithoke element is not fully used.The content that studies show that arithoke element in the leaf of Cynara scolymus L has reached about 1%, and the method for purification of therefore researching and developing arithoke element in the leaf of Cynara scolymus L is significant.
Arithoke element (cynarin) chemical name: 1,5-dicaffeoylquinic acid.Xln (dilute acetic acid), it is sweet to distinguish the flavor of.Molecular formula and molecular weight are respectively C
25H
24O
12With 516.46, fusing point is 225～227 ℃, and specific rotation [α] is-59 ° (c=2, methyl alcohol).Be slightly soluble in cold water, be soluble in the boiling water, be dissolved in Glacial acetic acid, alcohols.Its molecular structural formula is as follows:
That the arithoke element has is anti-oxidant, protect the liver, protect multiple biological activitys such as liver, reducing cholesterol, the most important thing is to have anti-HIV (human immunodeficiency virus) (Human immunodeficiency virus, HIV) activity.
At present, the patent of invention of existing multinomial method of purification about active substance in the leaf of Cynara scolymus L; As patent of invention: a kind of method of chlorogenic acid extracting (patent No.: 200810102863.4) just related to the method for purification of chlorogenic acid in the choke.
But existing method is not specially at extracting purifying arithoke element in the choke.Mostly be extraction at compounding substances about the patent of invention of extracting the arithoke element from leaf of Cynara scolymus L, wherein the content of arithoke element is not high, and the easy organic solvent of residual harmful.As patent of invention number be in 03150268 the Chinese invention patent " choke natural antioxidants and extracting method thereof ", choke dry powder is extracted the former extracting solution that obtains by Soxhlet, carry out chloroform, ethyl acetate, n-butanol extraction respectively, the propyl carbinol that obtains carries out Sephadex LH-20 column chromatography for separation mutually, obtain three different componentss, carry out the high-pressure liquid phase preparative column more respectively and separate, obtain 7 kinds of different natural antioxidantss.7 kinds of different natural antioxidantss that this invention obtains are respectively: 1-caffeoylquinic acids (1-CA), 3-caffeoylquinic acids (3-CA), luteolin-7-rutinoside (LTR), 1, and the 3-dicaffeoylquinic acid (1,3-DCA), 1, and the 5-dicaffeoylquinic acid (1,5-DCA), 3, the 4-dicaffeoylquinic acid (3,4-DCA), 3, the 5-dicaffeoylquinic acid (3,5-DCA).Patent of invention number is that U.S.'s patent of invention " Artichoke leaf extracts " water of 20040234674 or 80% methyl alcohol are as extracting solvent, after extracting solution is concentrated to proper volume, adding an amount of ethyl acetate/propyl carbinol extracts, obtain organic extraction phase, after behind the extraction phase concentrate drying, obtain to be rich in the coffee mesitoyl quinine acid material of 11-20%.
Summary of the invention
The technical problem to be solved in the present invention is, at the problem that does not have ad hoc approach purification arithoke element from choke in the prior art, provide a kind of from choke the method for purification arithoke element.
The technical solution adopted for the present invention to solve the technical problems is: the method for purification of arithoke element in a kind of choke is provided, comprises the following steps:
1) leaf of Cynara scolymus L is passed through organic solvent extraction, the gained extracting solution concentrates, and by the removal of impurity of depositing in water coarse filtration, carries out micro-filtration again, obtains dialysis liquid;
2) step 1) gained dialysis liquid is transferred to the pH value less than 6 with acid, it is carried out chromatographic separation, carry out the elutriant that wash-out obtains to contain the arithoke element with ethanol liquid;
3) with step 2) after the elutriant that contains the arithoke element that obtains concentrates, under 4-10 ℃, leave standstill crystallization, the crystal centrifugation gets the plain wet crystal of arithoke, and dry back obtains the plain thick product of arithoke.
The method of purification of arithoke element in the choke of the present invention also comprises step 4): the plain wet crystal recrystallization of arithoke that described step 3) is obtained obtains the plain product of arithoke.
The method of purification of arithoke element in the choke of the present invention, described step 2) in, described acid is hydrochloric acid soln or sulphuric acid soln; Described dialysis liquid transfers to the pH value and is 3-5.
The method of purification of arithoke element in the choke of the present invention, described step 2) in, the step of described chromatographic separation comprises, with step 2) in dialysis liquid behind the adjust pH by the macroporous adsorption resin chromatography post, wash with water to effluent liquid colourless earlier, be the ethanolic soln elution chromatography post of 10-18% with concentration of volume percent then, reclaim to such an extent that contain the elutriant of polyphenol; Be the ethanolic soln elution chromatography post of 50-70% with concentration of volume percent again, reclaim to such an extent that contain the elutriant of arithoke element.
The method of purification of arithoke element in the choke of the present invention, in the described step 4), described re-crystallization step comprises: the plain wet crystal of the arithoke of step 3) acquisition is added water dissolve again, add gac, stir the decolouring after-filtration, filtrate is left standstill recrystallization under 4-10 ℃, crystal centrifugation after drying obtains the plain product of arithoke.
The method of purification of arithoke element in the choke of the present invention, in the described step 1), described depositing in water coarse filtration step comprises: concentrated solution in the step 1) is added in the entry, leave standstill after stirring, depositing in water liquid filters, and following sediment abandons, and obtains filtered liquid.
The method of purification of arithoke element in the choke of the present invention, in the described step 1), the microfiltration membrane of described microfiltration process is a mineral membrane, the aperture of microfiltration membrane is 0.1-0.5 μ m.
The method of purification of arithoke element in the choke of the present invention, in the described step 1), described organic solvent is ethanol or acetone, described organic solvent concentration of volume percent is 50-70%.
The method of purification of arithoke element in the choke of the present invention, in described step 1) and the step 3), the condition of described concentration process is: temperature 60-70 ℃, pressure 50-300mmHg.
The method of purification of arithoke element in the choke of the present invention, in described step 3) and the step 4), the condition of described drying process is: temperature 60-70 ℃, pressure 10-50mmHg.
Method of purification of the present invention uses the ethanol of high volatility or acetone as extraction agent, extracts the arithoke element, organic solvent that can residual harmful in the product of gained.Implement the plain purity height of the contained arithoke of product that the present invention obtains, the plain product of the arithoke that step 4) obtains, the arithoke cellulose content can reach more than 80%; The plain thick product of the arithoke that step 3) obtains, the arithoke cellulose content is more than 20%, can offer the client of corresponding demand.The present invention also can obtain byproduct in leaching process, as step 2) the polyphenol elutriant that obtains can further handle and obtain the Polyphenols active substance.Integrated artistic of the present invention is simple to operate, and the production efficiency height is fit to suitability for industrialized production.
Description of drawings
The invention will be further described below in conjunction with drawings and Examples, in the accompanying drawing:
Fig. 1 is the schema of the method for purification of arithoke element in the choke of the present invention.
Embodiment
As shown in Figure 1, the step of the method for purification of arithoke element is in the choke of the present invention:
1. the fresh choke blade that will gather cuts into segment, is the organic solvent of 50-70% refluxing extraction 2 times under 60-80 ℃ condition with concentration of volume percent, collects and obtains its extracting solution;
2. with step 1 gained extracting solution concentrating under reduced pressure, reclaim organic solvent, obtain concentrated solution;
The concentrated solution that obtains of institute's step 2 add again be equivalent to 5 times of concentrated solution volumes water to carry out depositing in water; Depositing in water liquid obtains filtered liquid with filter paper filtering, and filter residue abandons;
4. the filter liquide that step 3 is obtained is that the mineral membrane of 0.1-0.5um carries out micro-filtration and handles with the aperture, and impurity such as the high molecular weight protein in the removal solution, tannin, pectin obtain limpid transparent mineral membrane dialysis liquid;
5. the mineral membrane dialysis liquid of step 4 is transferred between the pH to 3-5 with diluted acid, this liquid is the chromatography column of filler by macroporous adsorbent resin then, wash with water to effluent liquid colourless earlier, be that the ethanolic soln of 10-18% carries out the elution chromatography post with concentration of volume percent then, reclaim and contain the more weak polyphenol elutriant of polarity; Be that the ethanolic soln of 50%-70% carries out wash-out with concentration of volume percent again, reclaim the elutriant that contains the arithoke element;
6. step 5 gained is contained the elutriant concentrating under reduced pressure of arithoke element, concentrated solution leaves standstill crystallization under 4-10 ℃, and the crystal centrifugation gets the plain wet crystal of arithoke; The dry plain thick product of arithoke that gets of the plain wet crystal of arithoke;
7. add gac after the plain wet crystal of step 6 gained arithoke being dissolved in water, stir the decolouring after-filtration, filtrate is left standstill recrystallization under 4-10 ℃, and crystal centrifugation after drying obtains arithoke element product.
Plain thick product of the arithoke that the present invention obtains and the plain product of arithoke are by its content of liquid chromatograph check and analysis, shown in annex 1:
Annex 1
1 instrument and reagent
1.1 instrument
Waters 1525 high performance liquid chromatographs, Waters 2487 dual wavelength UV-detector,
Waters 515 pumps, Sartorius BS110S electronic balance
1.2 medicine and reagent
Acetonitrile: HPLC level, (Shanghai chemical reagents corporation of Chinese Medicine group)
Phosphoric acid: AR level, (chemical plant, Gansu Province, west, Shantou, Guangdong)
Ultrapure water: Millipore company
The plain reference substance (purity 95.0%) of arithoke: doctor Wang Mingfu of department of botany of Hong Kong University provides
2. experimental technique and result
2.1 chromatographic condition
Waters symmetry C-18 chromatographic column (5 μ m, 4.6 * 150mm)
Detect wavelength: λ=330nm
Flow velocity: 1.0mL/min, sample size: 10uL, column temperature: 25 ℃
Mobile phase A: acetonitrile, Mobile phase B: phosphoric acid/water [0.2: 99.8V/V]
Table 1:HPLC condition
Condition when table 1 is worked for liquid chromatograph, the scale parameter of moving phase second eyeball when wherein A% is liquid chromatograph work, the scale parameter of moving phase phosphoric acid/water when B% is liquid chromatograph work.
Embodiment 1:
1. the fresh blade of choke of gathering is cut into the segment about 5 centimetres.Take by weighing 1 kilogram of this blade, add 5 liters of concentration expressed in percentage by volumes and be 58% ethanolic soln, under 75 ℃ of conditions, stir and extracted 2 hours, time extracting solution of winning after the filtration.It is 58% ethanolic soln that filter residue adds 4 liters of concentration expressed in percentage by volumes again, under 75 ℃ of conditions, stir and extracted 1 hour, after the filtration extracting solution for the second time.Merge and extract the extracting solution that obtains for the first time and for the second time.
2. the extracting solution that step 1 is obtained is at 65 ℃, and being concentrated into relative density under the 200mmHg pressure condition is the concentrated solution of 1.2g/cm3 (measuring between 50-60 ℃).
3. the concentrated solution that step 2 is obtained adds the water that is equivalent to 5 times of concentrated solution volumes, leaves standstill behind the stirring and evenly mixing 6 hours, and with middling speed filter paper filtering depositing in water liquid, filter residue abandons, and obtains limpid filtered liquid.
4. the filtered liquid that step 3 is obtained carries out the micro-filtration processing with 0.3 μ m mineral membrane, and impurity such as the high molecular weight protein in the removal liquid, tannin, pectin obtain limpid mineral membrane dialyzate.
With the mineral membrane dialyzate of step 4 with dilute hydrochloric acid adjust pH to 3.5, with this liquid with 2.5BV/ hour flow velocity by with the AB-8 macroporous adsorbent resin being the chromatography column (5 centimetres of chromatography column diameters, 50 centimetres of height) of filler; To be washed till effluent liquid with 3BV/ hour flow velocity colourless for water then; Be 15% ethanolic soln elution chromatography post again with concentration of volume percent, elution speed is 2BV/ hour, collects to such an extent that contain the more weak polyphenol elutriant of polarity.Be 60% ethanolic soln elution chromatography post at last with concentration of volume percent, elution speed is 2BV/ hour, collects to such an extent that contain the elutriant of arithoke element.
6. the elutriant that will contain the arithoke element is at 65 ℃, and the 150mmHg pressure condition concentrates down, and being concentrated into relative density is 1.1g/cm
3(measuring between 50-60 ℃) places this concentrated solution under 4-10 ℃ then and leaves standstill crystallization, and the crystal centrifugation gets the plain wet crystal of arithoke.In 65 ℃, the pressure condition that vacuum tightness is lower than 30mmHg obtained the thick product of 15 grams down in dry 12 hours with the plain wet crystal of arithoke.The product of changing products are 22.6% through its arithoke cellulose content of HPLC check and analysis.
Embodiment 2:
1. the fresh blade of choke of gathering is cut into the segment about 5 centimetres.Take by weighing 1 kilogram of this blade, add 5 liters of concentration expressed in percentage by volumes and be 70% ethanolic soln, under 80 ℃ of conditions, stir and extracted 2 hours, time extracting solution of winning after the filtration.It is 70% ethanolic soln that filter residue adds 4 liters of concentration expressed in percentage by volumes again, under 80 ℃ of conditions, stir and extracted 1 hour, after the filtration extracting solution for the second time.Merge and extract the extracting solution that obtains for the first time and for the second time.
2. the extracting solution that step 1 is obtained is at 60 ℃, and being concentrated into relative density under the 50mmHg pressure condition is 1.2g/cm
3The concentrated solution of (between temperature 50-60 ℃, measuring).
3. the concentrated solution that step 2 is obtained adds the water that is equivalent to 5 times of concentrated solution volumes, leaves standstill behind the stirring and evenly mixing 6 hours, and with middling speed filter paper filtering depositing in water liquid, filter residue abandons.Obtain limpid filtered liquid.
4. the filtered liquid that step 3 is obtained carries out the micro-filtration processing with 0.1 μ m mineral membrane, and impurity such as the high molecular weight protein in the removal liquid, tannin, pectin obtain limpid mineral membrane dialyzate.
5. the mineral membrane dialyzate of step 4 is transferred pH to 3 with dilute sulphuric acid, with this liquid with 2.5BV/ hour flow velocity by with the AB-8 macroporous adsorbent resin being the chromatography column (chromatography column diameter 5cm, height 50cm) of filler; To be washed till effluent liquid with 3BV/ hour flow velocity colourless for water then; Be 10% ethanolic soln elution chromatography post again with concentration of volume percent, elution speed is 2BV/ hour, collects to such an extent that contain the more weak polyphenol elutriant of polarity.Be 50% ethanolic soln elution chromatography post at last with concentration of volume percent, elution speed is 2BV/ hour, collects to such an extent that contain the elutriant of arithoke element.
6. the elutriant that will contain the arithoke element is at 60 ℃, and the 50mmHg pressure condition concentrates down, and being concentrated into relative density is 1.1g/cm
3(measuring between 50-60 ℃) places this concentrated solution under 4 ℃ then and leaves standstill crystallization, and the crystal centrifugation gets the plain wet crystal of arithoke.
7. with the water dissolution of the plain wet crystal of the arithoke of step 6 acquisition with 5 times of weight, the adding mass percent is 1% gac, stir 15 minutes after-filtration of decolouring, filtrate is left standstill recrystallization under 10 ℃, the crystal centrifugation, in 60 ℃, 50mmHg pressure condition dry 12 hours down obtains the plain product of 10 gram arithokes.This product is 83.6% through its arithoke cellulose content of HPLC check and analysis.
Embodiment 3:
1. the fresh blade of choke of gathering is cut into the segment about 5 centimetres, taking by weighing 10 kilograms of these blades, to add 50 liters of concentration of volume percent be 50% acetone soln, stirs under 60 ℃ of conditions and extracted 2 hours, time extracting solution of winning after the filtration.It is 50% acetone soln that filter residue adds 40 liters of concentration expressed in percentage by volumes again, under 60 ℃ of conditions, stir and extracted 1 hour, after the filtration extracting solution for the second time.Merge and extract the extracting solution that obtains for the first time and for the second time.
2. the extracting solution that step 1 is obtained is at 70 ℃, and being concentrated into relative density under the 300mmHg pressure condition is 1.3g/cm
3The concentrated solution of (between 50-60 ℃, measuring).
3. the concentrated solution that step 2 is obtained adds the water that is equivalent to 5 times of concentrated solution volumes, leaves standstill behind the stirring and evenly mixing 6 hours, and with middling speed filter paper filtering depositing in water liquid, following sediment abandons.Obtain limpid filtered liquid.
4. the filtered liquid that step 3 is obtained carries out the micro-filtration processing with 0.5 μ m mineral membrane, and impurity such as the high molecular weight protein in the removal liquid, tannin, pectin obtain limpid mineral membrane dialyzate.
With the mineral membrane dialyzate of step 4 with dilute sulphuric acid adjust pH to 5, with this liquid with 2.5BV/ hour flow velocity by with the CAD-40 macroporous adsorbent resin being chromatography column (the chromatography column diameter 15cm of filler, height 100cm), to be washed till effluent liquid with 3BV/ hour flow velocity colourless for water then, be 18% ethanolic soln elution chromatography post again with concentration of volume percent, elution speed is 2BV/ hour, collects to such an extent that contain the more weak polyphenol elutriant of polarity.Be 70% ethanolic soln elution chromatography post at last with concentration of volume percent, elution speed is 2BV/ hour, collects to such an extent that contain the elutriant of arithoke element.
6. the elutriant that contains the arithoke element that step 5 is obtained is at 70 ℃, and the pressure condition of 300mmHg concentrates down, and being concentrated into relative density is 1.2g/cm
3(measuring between 50-60 ℃) places this concentrated solution under 10 ℃ then and leaves standstill crystallization, and the crystal centrifugation obtains the plain wet crystal of arithoke.
7. with the water dissolution of the plain wet crystal of the arithoke of step 6 acquisition with 5 times of weight, the adding mass percent concentration is 1% gac, stir 15 minutes after-filtration of decolouring, filtrate is left standstill recrystallization under 4 ℃, the crystal centrifugation, in 70 ℃, the 10mmHg pressure condition obtained the plain product of 30 gram arithokes down in dry 12 hours.This product is 80.8% through its arithoke cellulose content of HPLC check and analysis.
Claims (10)
1, the method for purification of arithoke element in a kind of choke is characterized in that, comprises the following steps:
1) leaf of Cynara scolymus L is passed through organic solvent extraction, the gained extracting solution concentrates, and by the removal of impurity of depositing in water coarse filtration, carries out micro-filtration again, obtains dialysis liquid;
2) step 1) gained dialysis liquid is transferred to the pH value less than 6 with acid, it is carried out chromatographic separation, carry out the elutriant that wash-out obtains to contain the arithoke element with ethanol liquid;
3) with step 2) after the elutriant that contains the arithoke element that obtains concentrates, under 4-10 ℃, leave standstill crystallization, the crystal centrifugation gets the plain wet crystal of arithoke, and dry back obtains the plain thick product of arithoke.
2, the method for purification of arithoke element in the choke according to claim 1 is characterized in that, also comprises step 4): the plain wet crystal recrystallization of arithoke that described step 3) is obtained obtains the plain product of arithoke.
3, the method for purification of arithoke element in the choke according to claim 1 is characterized in that described step 2) in, described acid is hydrochloric acid soln or sulphuric acid soln; Described dialysis liquid transfers to the pH value and is 3-5.
4, the method for purification of arithoke element in the choke according to claim 1, it is characterized in that, described step 2) in, the step of described chromatographic separation comprises, with step 2) in dialysis liquid behind the adjust pH by the macroporous adsorption resin chromatography post, washing with water to effluent liquid colourlessly earlier, is the ethanolic soln elution chromatography post of 10-18% with concentration of volume percent then, reclaims to such an extent that contain the elutriant of polyphenol; Be the ethanolic soln elution chromatography post of 50-70% with concentration of volume percent again, reclaim to such an extent that contain the elutriant of arithoke element.
5, the method for purification of arithoke element in the choke according to claim 2, it is characterized in that, in the described step 4), described re-crystallization step comprises: the plain wet crystal of the arithoke of step 3) acquisition is added water dissolve again, add gac, stir the decolouring after-filtration, filtrate is left standstill recrystallization under 4-10 ℃, crystal centrifugation after drying obtains the plain product of arithoke.
6, the method for purification of arithoke element in the choke according to claim 1 is characterized in that, in the described step 1), described depositing in water coarse filtration step comprises: concentrated solution in the step 1) is added in the entry, leave standstill after stirring, depositing in water liquid filters, following sediment abandons, and obtains filtered liquid.
7, the method for purification of arithoke element in the choke according to claim 1 is characterized in that, in the described step 1), the microfiltration membrane of described microfiltration process is a mineral membrane, and the aperture of microfiltration membrane is 0.1-0.5 μ m.
8, the method for purification of arithoke element in the choke according to claim 1 is characterized in that, in the described step 1), described organic solvent is ethanol or acetone, and described organic solvent concentration of volume percent is 50-70%.
9, the method for purification of arithoke element in the choke according to claim 1 is characterized in that, in described step 1) and the step 3), the condition of described concentration process is: temperature 60-70 ℃, and pressure 50-300mmHg.
10, the method for purification of arithoke element in the choke according to claim 2 is characterized in that, in described step 3) and the step 4), the condition of described drying process is: temperature 60-70 ℃, and pressure 10-50mmHg.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910107323XA CN101560155B (en) | 2009-05-13 | 2009-05-13 | Method for purifying cynarin in artichoke |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910107323XA CN101560155B (en) | 2009-05-13 | 2009-05-13 | Method for purifying cynarin in artichoke |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101560155A true CN101560155A (en) | 2009-10-21 |
| CN101560155B CN101560155B (en) | 2012-02-08 |
Family
ID=41219160
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910107323XA Expired - Fee Related CN101560155B (en) | 2009-05-13 | 2009-05-13 | Method for purifying cynarin in artichoke |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101560155B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102372634A (en) * | 2010-08-26 | 2012-03-14 | 苏州宝泽堂医药科技有限公司 | Preparation method for cynarin |
| CN106496033A (en) * | 2016-10-17 | 2017-03-15 | 汇美农业科技有限公司 | The extracting method of 1,5 dicaffeoylquinic acids in a kind of globe artichoke |
| CN107927779A (en) * | 2017-11-27 | 2018-04-20 | 爱可道生物科技有限公司 | A kind of beautifying face and moistering lotion oral liquid and preparation method thereof |
| CN108484402A (en) * | 2018-06-13 | 2018-09-04 | 爱可道生物科技有限公司 | The purifying technique of cynarin in a kind of globe artichoke |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1301721C (en) * | 2003-07-23 | 2007-02-28 | 昆明英之源农业科技开发有限公司 | Artichoke natural antioxidant and its extraction method |
| CN100496525C (en) * | 2004-04-02 | 2009-06-10 | 李嫦梅 | Technique of extracting acitve ingredient from roots, stems and leaves from artichoke and application |
-
2009
- 2009-05-13 CN CN200910107323XA patent/CN101560155B/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102372634A (en) * | 2010-08-26 | 2012-03-14 | 苏州宝泽堂医药科技有限公司 | Preparation method for cynarin |
| CN106496033A (en) * | 2016-10-17 | 2017-03-15 | 汇美农业科技有限公司 | The extracting method of 1,5 dicaffeoylquinic acids in a kind of globe artichoke |
| CN107927779A (en) * | 2017-11-27 | 2018-04-20 | 爱可道生物科技有限公司 | A kind of beautifying face and moistering lotion oral liquid and preparation method thereof |
| CN108484402A (en) * | 2018-06-13 | 2018-09-04 | 爱可道生物科技有限公司 | The purifying technique of cynarin in a kind of globe artichoke |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101560155B (en) | 2012-02-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109942380B (en) | Method for preparing cannabidiol by high-speed counter-current chromatography separation and purification | |
| CN102451235A (en) | Preparation method of olive leaf extract | |
| CN103435486A (en) | Novel method for preparing high-purity chlorogenic acid in honeysuckle | |
| CN101948501B (en) | Preparation method of hydroxyl asiaticoside | |
| CN102250195A (en) | Method for producing xanthoceraside | |
| CN101862385B (en) | Sanguisorba saponins and preparation method of sanguisorbin I | |
| CN102234245A (en) | Method for preparing sulforaphane | |
| CN101560155B (en) | Method for purifying cynarin in artichoke | |
| CN101348474A (en) | Method for preparing salvianolic acid B and tanshinol from Salvia miltiorrhiza stem | |
| CN101497637B (en) | Method for extracting high-purity scutellarin from breviscpini | |
| CN104211690B (en) | Method for separating and purifying mangiferin from aquilaria sinensis leaves | |
| CN103524591A (en) | Preparation method of effective components of triperygium wilfordii | |
| CN108218948A (en) | A kind of preparation method of Sodium Aescinate | |
| CN101255183A (en) | Method for extracting protodioscin from fenugreek | |
| CN102228488A (en) | Preparation of Lysimachia capillipes Hemsl total saponin | |
| CN104231011B (en) | Preparation method of verbascoside | |
| CN112336665A (en) | Sambucus williamsii extract and preparation method thereof | |
| CN102603854A (en) | Method for extracting asiaticoside from asiatic pennywort herb | |
| CN102329345A (en) | Method for extracting and purifying sarmentosin in Sedum sarmentosum Bunge | |
| CN102311467A (en) | Novel method for extracting and separating high purity tiliroside from edgeworthia chrysantha | |
| CN103739648A (en) | Preparation method for mussaendoside U | |
| CN101810317B (en) | Preparation method of canophyllic polyphenol and application thereof | |
| CN102603852A (en) | Preparation method of tripterine | |
| CN102648932A (en) | Method for extracting alfalfa saponin | |
| CN103288898B (en) | Neomangiferin reference substance extracts method for purifying and separating |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120208 Termination date: 20170513 |