CN115925768B - Method for extracting high-purity cyanidin galactoside from Aronia melanocarpa - Google Patents
Method for extracting high-purity cyanidin galactoside from Aronia melanocarpa Download PDFInfo
- Publication number
- CN115925768B CN115925768B CN202211478814.7A CN202211478814A CN115925768B CN 115925768 B CN115925768 B CN 115925768B CN 202211478814 A CN202211478814 A CN 202211478814A CN 115925768 B CN115925768 B CN 115925768B
- Authority
- CN
- China
- Prior art keywords
- microsphere
- aronia
- reverse phase
- phase chromatography
- purity
- 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.)
- Active
Links
- 240000005662 Aronia melanocarpa Species 0.000 title claims abstract description 48
- 235000007425 Aronia melanocarpa Nutrition 0.000 title claims abstract description 48
- -1 cyanidin galactoside Chemical class 0.000 title claims abstract description 32
- VEVZSMAEJFVWIL-UHFFFAOYSA-O cyanidin Natural products [O+]=1C2=CC(O)=CC(O)=C2C=C(O)C=1C1=CC=C(O)C(O)=C1 VEVZSMAEJFVWIL-UHFFFAOYSA-O 0.000 title claims abstract description 31
- 235000007336 cyanidin Nutrition 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 23
- 239000004005 microsphere Substances 0.000 claims abstract description 40
- 238000001179 sorption measurement Methods 0.000 claims abstract description 32
- 239000011347 resin Substances 0.000 claims abstract description 23
- 229920005989 resin Polymers 0.000 claims abstract description 23
- 239000002994 raw material Substances 0.000 claims abstract description 20
- 238000004108 freeze drying Methods 0.000 claims abstract description 15
- 238000004366 reverse phase liquid chromatography Methods 0.000 claims abstract description 14
- 239000002904 solvent Substances 0.000 claims abstract description 13
- 238000000746 purification Methods 0.000 claims abstract description 10
- 238000004458 analytical method Methods 0.000 claims abstract description 9
- 238000002425 crystallisation Methods 0.000 claims abstract description 8
- 230000008025 crystallization Effects 0.000 claims abstract description 8
- 238000002481 ethanol extraction Methods 0.000 claims abstract description 8
- 238000001953 recrystallisation Methods 0.000 claims abstract description 8
- 239000000945 filler Substances 0.000 claims abstract 9
- 238000011084 recovery Methods 0.000 claims abstract 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 46
- 238000012856 packing Methods 0.000 claims description 31
- 235000013399 edible fruits Nutrition 0.000 claims description 25
- 238000007710 freezing Methods 0.000 claims description 23
- 230000008014 freezing Effects 0.000 claims description 19
- 238000003795 desorption Methods 0.000 claims description 13
- 239000007788 liquid Substances 0.000 claims description 11
- 239000013078 crystal Substances 0.000 claims description 10
- 238000001914 filtration Methods 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 5
- 239000002352 surface water Substances 0.000 claims description 5
- 238000002137 ultrasound extraction Methods 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 4
- 239000012153 distilled water Substances 0.000 claims description 4
- 239000002245 particle Substances 0.000 claims description 4
- 239000010865 sewage Substances 0.000 claims description 4
- 239000000741 silica gel Substances 0.000 claims description 4
- 229910002027 silica gel Inorganic materials 0.000 claims description 4
- 230000010355 oscillation Effects 0.000 claims description 2
- 238000000859 sublimation Methods 0.000 claims description 2
- 230000008022 sublimation Effects 0.000 claims description 2
- 201000010099 disease Diseases 0.000 claims 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims 1
- 238000000605 extraction Methods 0.000 abstract description 10
- 238000011031 large-scale manufacturing process Methods 0.000 abstract description 2
- 229930002877 anthocyanin Natural products 0.000 description 9
- 235000010208 anthocyanin Nutrition 0.000 description 9
- 239000004410 anthocyanin Substances 0.000 description 9
- 150000004636 anthocyanins Chemical class 0.000 description 9
- 229930182830 galactose Natural products 0.000 description 6
- XFZJEEAOWLFHDH-UHFFFAOYSA-N (2R,2'R,3R,3'R,4R)-3,3',4',5,7-Pentahydroxyflavan(48)-3,3',4',5,7-pentahydroxyflavan Natural products C=12OC(C=3C=C(O)C(O)=CC=3)C(O)CC2=C(O)C=C(O)C=1C(C1=C(O)C=C(O)C=C1O1)C(O)C1C1=CC=C(O)C(O)=C1 XFZJEEAOWLFHDH-UHFFFAOYSA-N 0.000 description 5
- CWEZAWNPTYBADX-UHFFFAOYSA-N Procyanidin Natural products OC1C(OC2C(O)C(Oc3c2c(O)cc(O)c3C4C(O)C(Oc5cc(O)cc(O)c45)c6ccc(O)c(O)c6)c7ccc(O)c(O)c7)c8c(O)cc(O)cc8OC1c9ccc(O)c(O)c9 CWEZAWNPTYBADX-UHFFFAOYSA-N 0.000 description 5
- MOJZMWJRUKIQGL-FWCKPOPSSA-N Procyanidin C2 Natural products O[C@@H]1[C@@H](c2cc(O)c(O)cc2)Oc2c([C@H]3[C@H](O)[C@@H](c4cc(O)c(O)cc4)Oc4c3c(O)cc(O)c4)c(O)cc(O)c2[C@@H]1c1c(O)cc(O)c2c1O[C@@H]([C@H](O)C2)c1cc(O)c(O)cc1 MOJZMWJRUKIQGL-FWCKPOPSSA-N 0.000 description 5
- 229930182470 glycoside Natural products 0.000 description 5
- 229920002414 procyanidin Polymers 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 238000004140 cleaning Methods 0.000 description 4
- 239000003480 eluent Substances 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000010257 thawing Methods 0.000 description 3
- RKWHWFONKJEUEF-WVXKDWSHSA-O Idaein Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=CC2=C(O)C=C(O)C=C2[O+]=C1C1=CC=C(O)C(O)=C1 RKWHWFONKJEUEF-WVXKDWSHSA-O 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- YTMNONATNXDQJF-QSLGVYCOSA-N cyanidin 3-O-beta-D-galactoside chloride Chemical compound [Cl-].O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1OC1=CC2=C(O)C=C(O)C=C2[O+]=C1C1=CC=C(O)C(O)=C1 YTMNONATNXDQJF-QSLGVYCOSA-N 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 150000002338 glycosides Chemical group 0.000 description 2
- KZMACGJDUUWFCH-UHFFFAOYSA-O malvidin Chemical compound COC1=C(O)C(OC)=CC(C=2C(=CC=3C(O)=CC(O)=CC=3[O+]=2)O)=C1 KZMACGJDUUWFCH-UHFFFAOYSA-O 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 150000008442 polyphenolic compounds Chemical class 0.000 description 2
- 235000013824 polyphenols Nutrition 0.000 description 2
- HGVVOUNEGQIPMS-UHFFFAOYSA-N procyanidin Chemical compound O1C2=CC(O)=CC(O)=C2C(O)C(O)C1(C=1C=C(O)C(O)=CC=1)OC1CC2=C(O)C=C(O)C=C2OC1C1=CC=C(O)C(O)=C1 HGVVOUNEGQIPMS-UHFFFAOYSA-N 0.000 description 2
- ORTBMTXABUAMJS-CRTITRSXSA-N (4s,5r)-2-[2-(3,4-dihydroxyphenyl)-5,7-dihydroxychromenylium-3-yl]oxyoxane-3,4,5-triol;chloride Chemical compound [Cl-].OC1[C@@H](O)[C@H](O)COC1OC1=CC2=C(O)C=C(O)C=C2[O+]=C1C1=CC=C(O)C(O)=C1 ORTBMTXABUAMJS-CRTITRSXSA-N 0.000 description 1
- 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 1
- 241001444063 Aronia Species 0.000 description 1
- 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 1
- KUCVMQMKRICXJC-LMVHDODDSA-O Cyanidin 3-O-alpha-L-arabinopyranoside Natural products O([C@@H]1[C@H](O)[C@H](O)[C@@H](O)CO1)c1c(-c2cc(O)c(O)cc2)[o+]c2c(c(O)cc(O)c2)c1 KUCVMQMKRICXJC-LMVHDODDSA-O 0.000 description 1
- ORTBMTXABUAMJS-VGEDXCMYSA-N Cyanidin 3-arabinoside Chemical compound [Cl-].O[C@H]1[C@H](O)[C@H](O)CO[C@H]1OC1=CC2=C(O)C=C(O)C=C2[O+]=C1C1=CC=C(O)C(O)=C1 ORTBMTXABUAMJS-VGEDXCMYSA-N 0.000 description 1
- KUCVMQMKRICXJC-FBVAEJEDSA-O Cyanidin 3-arabinoside Natural products O([C@H]1[C@@H](O)[C@@H](O)[C@H](O)CO1)c1c(-c2cc(O)c(O)cc2)[o+]c2c(c(O)cc(O)c2)c1 KUCVMQMKRICXJC-FBVAEJEDSA-O 0.000 description 1
- GCPYCNBGGPHOBD-UHFFFAOYSA-N Delphinidin Natural products OC1=Cc2c(O)cc(O)cc2OC1=C3C=C(O)C(=O)C(=C3)O GCPYCNBGGPHOBD-UHFFFAOYSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 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 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- YKRGDOXKVOZESV-WRJNSLSBSA-N Paeoniflorin Chemical compound C([C@]12[C@H]3O[C@]4(O)C[C@](O3)([C@]1(C[C@@H]42)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)C)OC(=O)C1=CC=CC=C1 YKRGDOXKVOZESV-WRJNSLSBSA-N 0.000 description 1
- 235000004789 Rosa xanthina Nutrition 0.000 description 1
- 241000220222 Rosaceae Species 0.000 description 1
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 229930014669 anthocyanidin Natural products 0.000 description 1
- 235000008758 anthocyanidins Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 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 1
- 229940074393 chlorogenic acid Drugs 0.000 description 1
- 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 1
- 235000001368 chlorogenic acid Nutrition 0.000 description 1
- YTMNONATNXDQJF-UBNZBFALSA-N chrysanthemin Chemical compound [Cl-].O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC2=C(O)C=C(O)C=C2[O+]=C1C1=CC=C(O)C(O)=C1 YTMNONATNXDQJF-UBNZBFALSA-N 0.000 description 1
- 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 1
- 239000003086 colorant Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 235000007242 delphinidin Nutrition 0.000 description 1
- JKHRCGUTYDNCLE-UHFFFAOYSA-O delphinidin Chemical compound [O+]=1C2=CC(O)=CC(O)=C2C=C(O)C=1C1=CC(O)=C(O)C(O)=C1 JKHRCGUTYDNCLE-UHFFFAOYSA-O 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000004438 eyesight Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- NWKFECICNXDNOQ-UHFFFAOYSA-N flavylium Chemical compound C1=CC=CC=C1C1=CC=C(C=CC=C2)C2=[O+]1 NWKFECICNXDNOQ-UHFFFAOYSA-N 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 150000008195 galaktosides Chemical class 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 235000009584 malvidin Nutrition 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- YKRGDOXKVOZESV-UHFFFAOYSA-N paeoniflorin Natural products O1C(C)(C2(CC34)OC5C(C(O)C(O)C(CO)O5)O)CC3(O)OC1C24COC(=O)C1=CC=CC=C1 YKRGDOXKVOZESV-UHFFFAOYSA-N 0.000 description 1
- HKUHOPQRJKPJCJ-UHFFFAOYSA-N pelargonidin Natural products OC1=Cc2c(O)cc(O)cc2OC1c1ccc(O)cc1 HKUHOPQRJKPJCJ-UHFFFAOYSA-N 0.000 description 1
- 235000006251 pelargonidin Nutrition 0.000 description 1
- XVFMGWDSJLBXDZ-UHFFFAOYSA-O pelargonidin Chemical compound C1=CC(O)=CC=C1C(C(=C1)O)=[O+]C2=C1C(O)=CC(O)=C2 XVFMGWDSJLBXDZ-UHFFFAOYSA-O 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000014639 sexual reproduction Effects 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/10—Process efficiency
Landscapes
- Saccharide Compounds (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
本发明公开了一种从黑果腺肋花楸中高效提取矢车菊素半乳糖苷的方法，其特征在于包括如下步骤：1)原料预处理；2)乙醇提取；3)浓缩回收溶剂；4)大孔吸附树脂吸附及解析；5)浓缩回收溶剂；6)微球反相色谱填料纯化；7)低温浓缩结晶、重结晶；8)冷冻干燥。本发明方法通过二次大孔树脂吸附，并结合微球反相色谱填料纯化，可制备出纯度大于95％的矢车菊素半乳糖苷。本发明工艺过程简单，提取过程安全、温和，有利于在矢车菊素半乳糖苷的大规模生产中进行推广和应用。
The present invention discloses a method for efficiently extracting cyanidin galactoside from Aronia nigra, which is characterized by comprising the following steps: 1) raw material pretreatment; 2) ethanol extraction; 3) concentration and recovery of solvent; 4) macroporous adsorption resin adsorption and analysis; 5) concentration and recovery of solvent; 6) microsphere reverse phase chromatography filler purification; 7) low temperature concentration crystallization and recrystallization; 8) freeze drying. The method of the present invention can prepare cyanidin galactoside with a purity greater than 95% through secondary macroporous resin adsorption and combined with microsphere reverse phase chromatography filler purification. The process of the present invention is simple, and the extraction process is safe and mild, which is conducive to the promotion and application of cyanidin galactoside in large-scale production.
Description
Technical Field
The invention belongs to the technical field of extraction of plant active ingredients, and particularly relates to a method for extracting high-purity cyanidin galactoside from Aronia melanocarpa.
Background
Aronia melanocarpa is also called as Aronia melanocarpa, native to northeast of America, and is a perennial fallen leaf shrub of Aronia in Rosaceae. The Liaoning arid region institute in the 90 th century introduced one Aronia melanocarpa cultivar to northeast China, and the research of Aronia melanocarpa in China was started. Since introduction of China, the wide application of sexual reproduction and asexual propagation technology has been mastered through years of research, and research on cultivation, area test and the like has been completed successively, and the current planting areas are mainly concentrated in eastern and middle areas of China. The Aronia melanocarpa is mainly of three values of medical use, edibility and ornamental value. Aronia melanocarpa is rich in anthocyanin, procyanidine, flavone, chlorogenic acid, dietary fiber, etc. In 2018, the national health committee issued "bulletin on new food raw materials of Aronia melanocarpa", which shows that Aronia melanocarpa and its products have potential to promote health, and have potential market prospects in industries such as health care products, natural food additives, cosmetics, and the like.
Anthocyanin is a glycoside form of anthocyanin, which is widely present in the cell fluid of flowers, fruits, stems, leaves and root organs of plants, and makes the plants present different colors from red, purplish red to blue. The anthocyanin mainly comprises 6 non-glycoside of pelargonidin, cyanidin, delphinidin, paeoniflorin, petuniin and malvidin. Researches show that anthocyanin has the effects of resisting oxidation, resisting fatigue, resisting inflammation, resisting tumor, enhancing eyesight and the like, and is widely applied to the fields of foods, biology, medicines and the like at present. The Aronia melanocarpa fruit is rich in polyphenols, and the polyphenols content is the highest in the currently known fruits, wherein the anthocyanin content can be one fourth of the total phenols. The Aronia melanocarpa anthocyanin mainly comprises 4 anthocyanidins including cyanidin-3-O-galactoside, cyanidin-3-arabinoside, cyanidin-3-glucoside and cyanidin-3-xyloside, wherein the cyanidin galactoside accounts for 68.68% at maximum.
The cyanidin-3-O-galactoside (Cyanidin-galactoside) is also called cyanidin galactoside, and has various physiological and biochemical effects of protecting blood vessel, resisting blood coagulation, etc. Therefore, the Aronia melanocarpa fruit is used as a raw material to extract, separate and purify the procyanidin galactose glycoside so as to prepare the high-purity procyanidin galactose glycoside product, and the Aronia melanocarpa fruit has wide market prospect. At present, most of the existing researches utilize conventional methods, such as solvent extraction method, ultrasonic assisted extraction method and the like, to extract anthocyanin from Aronia melanocarpa, and reports on extraction, separation and purification of procyanidin galactoside are seen.
Disclosure of Invention
Aiming at the problems, the invention provides a method for extracting high-purity cyanidin galactoside from Aronia melanocarpa, the method can prepare the cyanidin galactoside with the purity of more than 95 percent by secondary macroporous resin adsorption and microsphere reversed phase chromatographic packing purification.
The invention is realized by the following technical scheme.
A method for efficiently extracting cyanidin galactoside from Aronia melanocarpa is characterized by comprising the following steps:
1) Pretreatment of raw materials
Picking full and pest-free Aronia melanocarpa fruits as raw materials, cleaning, draining surface water, quick-freezing, and preserving at low temperature below-18 for later use;
2) Ethanol extraction
After the raw materials in the step (1) are subjected to microwave thawing-slow freezing cycle treatment for 2-3 times, adding ethanol for ultrasonic auxiliary extraction, and then centrifugally filtering and collecting an extracting solution for later use;
3) Concentrating and recovering solvent
4) Adsorption and analysis by macroporous adsorption resin
Adsorbing the concentrated solution in the step (3) by using macroporous adsorption resin I, then carrying out ethanol stepwise desorption, then adsorbing by using macroporous adsorption resin II, then carrying out ethanol stepwise desorption, and collecting the finally obtained desorption solution for later use;
5) Concentrating and recovering solvent
6) Microsphere reversed phase chromatography packing purification
Adding the microsphere reversed phase chromatographic packing into the concentrated solution in the step (5), stirring for 10-30 min, standing to remove the lower layer of settled microsphere reversed phase chromatographic packing, eluting the microsphere reversed phase chromatographic packing, and collecting the eluent to obtain a purified solution;
7) Concentrating, crystallizing and recrystallizing at low temperature
Sequentially carrying out low-temperature concentration crystallization and recrystallization on the purified solution obtained in the step (6), and collecting precipitated crystals for later use;
8) Freeze drying
And (3) freeze-drying the crystal obtained in the step (7) to obtain the cyanidin galactoside.
Preferably, in the step (2), the feed liquid ratio of the Aronia melanocarpa fruits to the ethanol is 1:6-12 kg/L.
Preferably, in the step (2), the power of the ultrasonic wave is 150-200 w, and the ultrasonic wave extraction time is 15-30 min.
Preferably, in the step (2), the specific parameters of microwave thawing-slow thawing are that frozen Aronia melanocarpa fruits are placed in a container, then thawed for 2-5 min under the microwave power of 0.2-2.5 kw/kg, and then the thawed Aronia melanocarpa fruits are placed in the condition of-12 to-8 ℃ for slow thawing for 4-10 h.
Preferably, in the step (4), the macroporous adsorption resin I type is D101 type, and the macroporous adsorption resin II type is AB-8 type.
Preferably, in the step (6), the microsphere reverse phase chromatographic packing is porous silica gel, and the particle size is 18-30 μm.
Preferably, in the step (6), the microsphere reversed phase chromatographic packing is pretreated before being used, namely, the microsphere reversed phase chromatographic packing is mixed with sewage ethanol according to the feed liquid ratio of 1:3-5 kg/L, then the mixture is placed in ultrasonic oscillation for treatment for 10-15 min, and then the mixture is filtered and separated to obtain the microsphere reversed phase chromatographic packing, and the microsphere reversed phase chromatographic packing is continuously leached by distilled water and then dried at low temperature.
Preferably, in the step (8), the parameters of freeze drying are that the pre-freezing temperature is-55 to-45 ℃ and the time is 4 to 6 hours, the sublimation drying temperature is-35 to-30 ℃ and the time is 22 to 24 hours, the analysis temperature is 30 ℃ and the time is 16 to 20 hours, the vacuum degree of a freeze dryer is 1.0Pa, and the cold trap temperature is-55 to-45 ℃.
The invention has the beneficial effects that:
1) According to the application, the picked Aronia melanocarpa fruits are quick-frozen and stored at low temperature, so that anthocyanin components in the Aronia melanocarpa fruits can be furthest kept from being damaged and lost, and the high-purity cyanidin galactoside can be obtained; in addition, the microwave thawing-slow freezing circulation treatment is used in the ethanol extraction, and the cell structure of the Aronia melanocarpa fruit can be damaged through repeated thawing and freezing, so that the dissolution of active ingredients in cells is facilitated, the ethanol extraction yield is improved, and the extraction effect is further enhanced through an ultrasonic-assisted means.
2) The method can prepare the cyanidin galactoside with the purity of more than 95 percent by secondary macroporous resin adsorption and combination with microsphere reversed phase chromatographic packing purification, low-temperature concentration crystallization and recrystallization.
3) The method has simple technical process, safe and mild extraction process, and is favorable for popularization and application in large-scale production of the cyanidin galactoside.
Drawings
FIG. 1 is a liquid chromatogram of a solvent blank;
FIG. 2 is a liquid chromatogram of a cyanidin galactosyl sample.
Detailed Description
The present invention will be further described with reference to the following specific embodiments, which are given by way of illustration only, but the scope of the invention is not limited thereto, and all equivalents of the invention within the spirit of the invention will fall within the scope of the invention by those skilled in the art.
Example 1
A method for efficiently extracting cyanidin galactoside from Aronia melanocarpa comprises the following steps:
1) Pretreatment of raw materials
Picking full and pest-free Aronia melanocarpa fruits as raw materials, cleaning, draining surface water, quick-freezing, and preserving at low temperature below-18 for later use;
2) Ethanol extraction
After the raw materials in the step (1) are subjected to microwave thawing-slow freezing cycle treatment for 2-3 times, adding ethanol for ultrasonic auxiliary extraction, and then centrifugally filtering and collecting an extracting solution for later use;
3) Concentrating and recovering solvent
4) Adsorption and analysis by macroporous adsorption resin
Adsorbing the concentrated solution in the step (3) by adopting macroporous adsorption resin I, then carrying out ethanol stepwise desorption, then carrying out adsorption by adopting macroporous adsorption resin II, carrying out ethanol stepwise desorption, and collecting the finally obtained desorption solution for later use;
5) Concentrating and recovering solvent
6) Microsphere reversed phase chromatography packing purification
Adding the microsphere reversed phase chromatographic packing into the concentrated solution in the step (5), stirring for 10-30 min, standing to remove the lower layer of settled microsphere reversed phase chromatographic packing, eluting the microsphere reversed phase chromatographic packing, and collecting the eluent to obtain a purified solution;
7) Concentrating, crystallizing and recrystallizing at low temperature
Sequentially carrying out low-temperature concentration crystallization and recrystallization on the purified solution obtained in the step (6), and collecting precipitated crystals for later use;
8) Freeze drying
And (3) freeze-drying the crystal obtained in the step (7) to obtain the cyanidin galactoside.
Example 2
A method for efficiently extracting cyanidin galactose glycoside from Aronia melanocarpa comprises the following steps:
1) Pretreatment of raw materials
Picking full and pest-free Aronia melanocarpa fruits as raw materials, cleaning, draining surface water, quick-freezing, and preserving at low temperature below-18 for later use;
2) Ethanol extraction
After the raw materials in the step (1) are subjected to microwave thawing-slow freezing circulation treatment for 2-3 times, wherein the specific parameters of microwave thawing-slow freezing are that frozen Aronia melanocarpa fruits are placed in a container, then thawed for 2-5 min under the microwave power of 0.2-2.5 kw/kg, and then the thawed Aronia melanocarpa fruits are placed in the condition of-12 to-8 ℃ for slow freezing for 4-10 h;
After the raw materials are subjected to microwave thawing-slow freezing cycle treatment for 2-3 times, adding ethanol according to the feed liquid ratio of the Aronia melanocarpa fruits to the ethanol of 1:6-12 kg/L, performing ultrasonic auxiliary extraction under the conditions that the power of ultrasonic waves is 150-200 w and the ultrasonic extraction time is 15-30 min, and then centrifugally filtering and collecting an extracting solution for later use;
3) Concentrating and recovering solvent
4) Adsorption and analysis by macroporous adsorption resin
Adsorbing the concentrated solution in the step (3) by using D101 type macroporous adsorption resin, then carrying out ethanol stepwise desorption, then carrying out adsorption by using AB-8 type macroporous adsorption resin, carrying out ethanol stepwise desorption, and collecting the finally obtained desorption solution for later use;
5) Concentrating and recovering solvent
6) Microsphere reversed phase chromatography packing purification
Selecting porous silica gel with the particle size of 18-30 μm as microsphere reversed phase chromatographic packing, firstly mixing the microsphere reversed phase chromatographic packing with sewage ethanol according to the feed liquid ratio of 1:3-5, then placing the mixture in ultrasonic vibration for treatment for 10-15 min, and then filtering and separating to obtain the microsphere reversed phase chromatographic packing, and continuously leaching the microsphere reversed phase chromatographic packing with distilled water and then drying at low temperature;
Adding the microsphere reversed phase chromatographic packing into the concentrated solution in the step (5), stirring for 10-30 min, standing to remove the lower layer of settled microsphere reversed phase chromatographic packing, eluting the microsphere reversed phase chromatographic packing, and collecting the eluent to obtain a purified solution;
7) Concentrating, crystallizing and recrystallizing at low temperature
Sequentially carrying out low-temperature concentration crystallization and recrystallization on the purified solution obtained in the step (6), and collecting precipitated crystals for later use;
8) Freeze drying
And (3) freeze-drying the crystal obtained in the step (7) to obtain the cyaniding galactoside, wherein the parameters of freeze-drying are that the pre-freezing temperature is-55 to-45 ℃ and the time is 4-6 hours, the sublimation-drying temperature is-35 to-30 ℃ and the time is 22-24 hours, the analysis temperature is 30 ℃ and the time is 16-20 hours, the vacuum degree of a freeze dryer is 1.0Pa, and the cold trap temperature is-55 to-45 ℃.
Example 3
A method for efficiently extracting cyanidin galactose glycoside from Aronia melanocarpa comprises the following steps:
1) Pretreatment of raw materials
Picking full and pest-free Aronia melanocarpa fruits as raw materials, cleaning, draining surface water, quick-freezing, and preserving at low temperature below-18 for later use;
2) Ethanol extraction
After the raw materials in the step (1) are subjected to microwave thawing-slow freezing circulation treatment for 2 times, wherein the specific parameters of microwave thawing-slow freezing are that frozen Aronia melanocarpa fruits are placed in a container, then thawed for 5min under the microwave power of 0.5kw/kg, and then the thawed Aronia melanocarpa fruits are placed in a condition of-10 ℃ for slow freezing for 10h;
After the raw materials are subjected to microwave thawing-slow freezing circulation treatment for 2 times, adding ethanol according to the feed liquid ratio of the Aronia melanocarpa fruits to the ethanol of 1:10kg/L, performing ultrasonic auxiliary extraction under the conditions that the power of ultrasonic waves is 150-200 w and the ultrasonic extraction time is 15min, and centrifuging, filtering and collecting the extracting solution for later use;
3) Concentrating and recovering solvent
4) Adsorption and analysis by macroporous adsorption resin
Adsorbing the concentrated solution in the step (3) by using D101 type macroporous adsorption resin, then carrying out ethanol stepwise desorption, then carrying out adsorption by using AB-8 type macroporous adsorption resin, carrying out ethanol stepwise desorption, and collecting the finally obtained desorption solution for later use;
5) Concentrating and recovering solvent
6) Microsphere reversed phase chromatography packing purification
Selecting porous silica gel with the particle size of 20 mu m as microsphere reversed phase chromatographic packing, firstly mixing the microsphere reversed phase chromatographic packing with sewage ethanol according to the feed liquid ratio of 1:5, then placing the mixture in ultrasonic vibration for treatment for 10min, filtering and separating to obtain the microsphere reversed phase chromatographic packing, continuously eluting with distilled water, and drying at low temperature;
Adding the microsphere reversed phase chromatographic packing into the concentrated solution in the step (5), stirring for 30min, standing to remove the lower layer of settled microsphere reversed phase chromatographic packing, eluting the microsphere reversed phase chromatographic packing, and collecting the eluent to obtain a purified solution;
7) Concentrating, crystallizing and recrystallizing at low temperature
Sequentially carrying out low-temperature concentration crystallization and recrystallization on the purified solution obtained in the step (6), and collecting precipitated crystals for later use;
8) Freeze drying
Freeze-drying the crystal obtained in the step (7), wherein the parameters of freeze-drying are that the pre-freezing temperature is-55 to-45 ℃ and the time is 4-6 hours, the sublimation-drying temperature is-35 to-30 ℃ and the time is 22-24 hours, the analysis temperature is 30 ℃ and the time is 16-20 hours, the vacuum degree of a freeze dryer is 1.0Pa, the cold trap temperature is-55 to-45 ℃, and finally, 6.09g of the procyanidin galactose is prepared, and the purity of the prepared procyanidin galactose is detected by adopting liquid chromatography, wherein the detection conditions are that:
Chromatographic column C18 ME, 5um, 100A, 4.6X250 mm;
A detector, wherein UV is 200-600 nm;
the detection wavelength is 520nm;
The mobile phase is as follows:
| Time (min) | Aacetonitrile | B0.2% phosphoric acid water |
| 0 | 13 | 87 |
| 15 | 13 | 87 |
The liquid chromatogram of the obtained cyanidin galactoside is shown in figure 2, and the purity of the cyanidin galactoside reaches 95.62% through calculation.
Claims (6)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211478814.7A CN115925768B (en) | 2022-11-23 | 2022-11-23 | Method for extracting high-purity cyanidin galactoside from Aronia melanocarpa |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211478814.7A CN115925768B (en) | 2022-11-23 | 2022-11-23 | Method for extracting high-purity cyanidin galactoside from Aronia melanocarpa |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115925768A CN115925768A (en) | 2023-04-07 |
| CN115925768B true CN115925768B (en) | 2024-12-17 |
Family
ID=86648142
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211478814.7A Active CN115925768B (en) | 2022-11-23 | 2022-11-23 | Method for extracting high-purity cyanidin galactoside from Aronia melanocarpa |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115925768B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108409703A (en) * | 2018-06-07 | 2018-08-17 | 中国农业大学 | A method for separating anthocyanins with anti-aging effect by simulating moving bed |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8153866B2 (en) * | 2007-06-21 | 2012-04-10 | Suntava Llc | Corn inbreds like FAR045 and hybrids thereof |
| KR20090049148A (en) * | 2007-11-13 | 2009-05-18 | (주)마이크로허브 | Separation of Anthocyanins and Polyphenols as High Purity from Black Chokeberry Fruit |
| KR20130090485A (en) * | 2012-02-06 | 2013-08-14 | 이강수 | Anthocyanidin and anthocyanins maded from omija pigment |
| CN107325138B (en) * | 2016-11-08 | 2020-04-28 | 东北林业大学 | A method for extracting, separating and purifying four main anthocyanins from blackcurrant pomace |
| KR20190053607A (en) * | 2017-11-10 | 2019-05-20 | 충청북도 단양군 | Method for preparing Aronia extract comprising high pressure homogenization process and pharmaceutical composition for improving immunity comprising Aronia extract prepared thereby as active ingredients |
| CN112089037A (en) * | 2020-09-16 | 2020-12-18 | 江南大学 | A method for preparing berry product rich in type A procyanidin |
| CN115057900B (en) * | 2022-07-26 | 2023-08-08 | 山东省农业科学院 | A kind of method for extracting anthocyanin from Aronia nigra |
-
2022
- 2022-11-23 CN CN202211478814.7A patent/CN115925768B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108409703A (en) * | 2018-06-07 | 2018-08-17 | 中国农业大学 | A method for separating anthocyanins with anti-aging effect by simulating moving bed |
Non-Patent Citations (2)
| Title |
|---|
| "黑果腺肋花楸花色苷提取分离纯化及微胶囊化的研究";唐璐;《中国优秀硕士学位论文全文数据库（电子期刊） 工程科技Ⅰ辑》;20200315(第03期);第B024-317页 * |
| 赵余庆.《中药及天然产物提取制备关键技术》.中国医药科技出版社,2012,第132-133页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115925768A (en) | 2023-04-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109810201B (en) | Ultrasonic wave composite acidic water extraction method for cordyceps polysaccharide and cordycepin in cordyceps militaris | |
| CN103159864B (en) | Five kinds of polysaccharide that a kind of separation purification method of lycium ruthenicum polysaccharide and separation obtain | |
| CN101265252A (en) | Vaccinium anthocyanin and its separation and purification method | |
| CN105153091B (en) | Method for improving yield of dihydromyricetin in ampelopsis grossedentata | |
| CN110183542B (en) | A kind of extraction method of poplar Phellinus polysaccharide | |
| CN107686473B (en) | A kind of joint preparation method of polysaccharide and polyphenol of purple cabbage | |
| CN102212144B (en) | A method for preparing pure polysaccharide from alfalfa hay | |
| CN102219865A (en) | Preparation method of cherokee rose polysaccharide derivatives with antitumor activity | |
| CN107970270A (en) | A kind of method of step by step arithmetic general flavone and polysaccharide from folium cortex eucommiae | |
| CN104764846B (en) | A kind of extraction, purification, method of identification anthocyanidin from fresh leaf of Camelliae sinensis | |
| CN113024679B (en) | A kind of method for extracting selenium polysaccharides and polyphenols in selenium-enriched Moringa seeds | |
| CN105524183B (en) | A kind of extraction method of water chestnut polysaccharide | |
| CN116531431A (en) | Extraction, separation and purification method of total flavonoids in hawthorn seeds | |
| CN101229335A (en) | Method for Enzymatically Preparing Smilax Smilax Total Saponins Extract | |
| CN108567836A (en) | A method of combined extraction separation flavones and polysaccharide from hawthorn skin slag | |
| CN110882285A (en) | Efficient preparation method of active substances in Phellinus linteus | |
| CN108467345B (en) | Method for extracting nervonic acid from garlic fruits and nervonic acid clathrate compound | |
| CN115925768B (en) | Method for extracting high-purity cyanidin galactoside from Aronia melanocarpa | |
| CN106008647A (en) | Novel extraction method of momordica saponins | |
| WO2019184025A1 (en) | Method for preparing polyarabogalacturonic acid by using dried tangerine peel | |
| CN118667042A (en) | Polygonatum sibiricum polysaccharide extraction process | |
| CN107988280A (en) | The method that isothiocyanates high sterling is obtained through refining from brassicaceous vegetable seed | |
| CN111718428A (en) | A kind of method that utilizes Dendrobium officinale fermentation broth to prepare water-soluble polysaccharide | |
| CN111171112A (en) | Enzymatic extraction method of bitter melon seed protein | |
| CN114249666B (en) | Extraction method of betaine in phoenix tree leaves |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |