KR20230003852A - Method for extracting and separating flavonoid from natural product using ionic liquids - Google Patents
Method for extracting and separating flavonoid from natural product using ionic liquids Download PDFInfo
- Publication number
- KR20230003852A KR20230003852A KR1020210085435A KR20210085435A KR20230003852A KR 20230003852 A KR20230003852 A KR 20230003852A KR 1020210085435 A KR1020210085435 A KR 1020210085435A KR 20210085435 A KR20210085435 A KR 20210085435A KR 20230003852 A KR20230003852 A KR 20230003852A
- Authority
- KR
- South Korea
- Prior art keywords
- methylimidazolium
- ionic liquid
- butyl
- flavonoids
- solvent
- Prior art date
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- 229930003935 flavonoid Natural products 0.000 title claims abstract description 92
- 150000002215 flavonoids Chemical class 0.000 title claims abstract description 92
- 235000017173 flavonoids Nutrition 0.000 title claims abstract description 92
- 239000002608 ionic liquid Substances 0.000 title claims abstract description 73
- 229930014626 natural product Natural products 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 title claims description 45
- 238000000605 extraction Methods 0.000 claims abstract description 53
- 239000002904 solvent Substances 0.000 claims abstract description 53
- 239000002244 precipitate Substances 0.000 claims abstract description 37
- 238000001953 recrystallisation Methods 0.000 claims abstract description 25
- 239000012296 anti-solvent Substances 0.000 claims abstract description 13
- 238000000926 separation method Methods 0.000 claims abstract description 11
- 238000001914 filtration Methods 0.000 claims abstract description 9
- -1 oxazolium cation Chemical class 0.000 claims description 68
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 42
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 27
- 239000000284 extract Substances 0.000 claims description 25
- 238000002137 ultrasound extraction Methods 0.000 claims description 19
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 13
- ABFDKXBSQCTIKH-UHFFFAOYSA-M 1-ethylpyridin-1-ium;bromide Chemical compound [Br-].CC[N+]1=CC=CC=C1 ABFDKXBSQCTIKH-UHFFFAOYSA-M 0.000 claims description 10
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- KYCQOKLOSUBEJK-UHFFFAOYSA-M 1-butyl-3-methylimidazol-3-ium;bromide Chemical compound [Br-].CCCCN1C=C[N+](C)=C1 KYCQOKLOSUBEJK-UHFFFAOYSA-M 0.000 claims description 7
- FHDQNOXQSTVAIC-UHFFFAOYSA-M 1-butyl-3-methylimidazol-3-ium;chloride Chemical compound [Cl-].CCCCN1C=C[N+](C)=C1 FHDQNOXQSTVAIC-UHFFFAOYSA-M 0.000 claims description 6
- GWQYPLXGJIXMMV-UHFFFAOYSA-M 1-ethyl-3-methylimidazol-3-ium;bromide Chemical compound [Br-].CCN1C=C[N+](C)=C1 GWQYPLXGJIXMMV-UHFFFAOYSA-M 0.000 claims description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 6
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 6
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 6
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 6
- 150000001768 cations Chemical class 0.000 claims description 6
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 5
- 150000001450 anions Chemical class 0.000 claims description 5
- 235000019253 formic acid Nutrition 0.000 claims description 5
- 150000003949 imides Chemical class 0.000 claims description 5
- URVSXZLUUCVGQM-UHFFFAOYSA-M 1-methyl-3-octylimidazol-1-ium;bromide Chemical compound [Br-].CCCCCCCCN1C=C[N+](C)=C1 URVSXZLUUCVGQM-UHFFFAOYSA-M 0.000 claims description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 4
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- NJMWOUFKYKNWDW-UHFFFAOYSA-N 1-ethyl-3-methylimidazolium Chemical compound CCN1C=C[N+](C)=C1 NJMWOUFKYKNWDW-UHFFFAOYSA-N 0.000 claims description 3
- RAXXELZNTBOGNW-UHFFFAOYSA-O Imidazolium Chemical compound C1=C[NH+]=CN1 RAXXELZNTBOGNW-UHFFFAOYSA-O 0.000 claims description 3
- JUJWROOIHBZHMG-UHFFFAOYSA-O pyridinium Chemical compound C1=CC=[NH+]C=C1 JUJWROOIHBZHMG-UHFFFAOYSA-O 0.000 claims description 3
- CCEYVAPCYYFRMO-UHFFFAOYSA-N 1,3-didecyl-2-methylimidazol-1-ium Chemical compound CCCCCCCCCCN1C=C[N+](CCCCCCCCCC)=C1C CCEYVAPCYYFRMO-UHFFFAOYSA-N 0.000 claims description 2
- KSOGGGZFEJTGPZ-UHFFFAOYSA-M 1-butyl-2,3-dimethylimidazol-3-ium;trifluoromethanesulfonate Chemical compound [O-]S(=O)(=O)C(F)(F)F.CCCC[N+]=1C=CN(C)C=1C KSOGGGZFEJTGPZ-UHFFFAOYSA-M 0.000 claims description 2
- ICIVTHOGIQHZRY-UHFFFAOYSA-N 1-butyl-3-methylimidazol-3-ium;cyanoiminomethylideneazanide Chemical compound [N-]=C=NC#N.CCCCN1C=C[N+](C)=C1 ICIVTHOGIQHZRY-UHFFFAOYSA-N 0.000 claims description 2
- PUHVBRXUKOGSBC-UHFFFAOYSA-M 1-butyl-3-methylimidazol-3-ium;methanesulfonate Chemical compound CS([O-])(=O)=O.CCCC[N+]=1C=CN(C)C=1 PUHVBRXUKOGSBC-UHFFFAOYSA-M 0.000 claims description 2
- FRZPYEHDSAQGAS-UHFFFAOYSA-M 1-butyl-3-methylimidazol-3-ium;trifluoromethanesulfonate Chemical compound [O-]S(=O)(=O)C(F)(F)F.CCCC[N+]=1C=CN(C)C=1 FRZPYEHDSAQGAS-UHFFFAOYSA-M 0.000 claims description 2
- HXMUPILCYSJMLQ-UHFFFAOYSA-M 1-ethyl-3-methylimidazol-3-ium;4-methylbenzenesulfonate Chemical compound CC[N+]=1C=CN(C)C=1.CC1=CC=C(S([O-])(=O)=O)C=C1 HXMUPILCYSJMLQ-UHFFFAOYSA-M 0.000 claims description 2
- VRFOKYHDLYBVAL-UHFFFAOYSA-M 1-ethyl-3-methylimidazol-3-ium;ethyl sulfate Chemical compound CCOS([O-])(=O)=O.CCN1C=C[N+](C)=C1 VRFOKYHDLYBVAL-UHFFFAOYSA-M 0.000 claims description 2
- IXLWEDFOKSJYBD-UHFFFAOYSA-M 1-ethyl-3-methylimidazol-3-ium;methanesulfonate Chemical compound CS([O-])(=O)=O.CC[N+]=1C=CN(C)C=1 IXLWEDFOKSJYBD-UHFFFAOYSA-M 0.000 claims description 2
- NQRYJNQNLNOLGT-UHFFFAOYSA-O Piperidinium(1+) Chemical compound C1CC[NH2+]CC1 NQRYJNQNLNOLGT-UHFFFAOYSA-O 0.000 claims description 2
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 claims description 2
- KXKVLQRXCPHEJC-UHFFFAOYSA-N acetic acid trimethyl ester Natural products COC(C)=O KXKVLQRXCPHEJC-UHFFFAOYSA-N 0.000 claims description 2
- IEFUHGXOQSVRDQ-UHFFFAOYSA-N bis(trifluoromethylsulfonyl)azanide;1-methyl-1-propylpiperidin-1-ium Chemical compound CCC[N+]1(C)CCCCC1.FC(F)(F)S(=O)(=O)[N-]S(=O)(=O)C(F)(F)F IEFUHGXOQSVRDQ-UHFFFAOYSA-N 0.000 claims description 2
- ZDVNJIBUDKGGGM-UHFFFAOYSA-N bis(trifluoromethylsulfonyl)azanide;diethyl(methyl)sulfanium Chemical compound CC[S+](C)CC.FC(F)(F)S(=O)(=O)[N-]S(=O)(=O)C(F)(F)F ZDVNJIBUDKGGGM-UHFFFAOYSA-N 0.000 claims description 2
- BLODSRKENWXTLO-UHFFFAOYSA-N bis(trifluoromethylsulfonyl)azanide;triethylsulfanium Chemical compound CC[S+](CC)CC.FC(F)(F)S(=O)(=O)[N-]S(=O)(=O)C(F)(F)F BLODSRKENWXTLO-UHFFFAOYSA-N 0.000 claims description 2
- MKHFCTXNDRMIDR-UHFFFAOYSA-N cyanoiminomethylideneazanide;1-ethyl-3-methylimidazol-3-ium Chemical compound [N-]=C=NC#N.CCN1C=C[N+](C)=C1 MKHFCTXNDRMIDR-UHFFFAOYSA-N 0.000 claims description 2
- YNAVUWVOSKDBBP-UHFFFAOYSA-O morpholinium Chemical compound [H+].C1COCCN1 YNAVUWVOSKDBBP-UHFFFAOYSA-O 0.000 claims description 2
- LAGQNGWYNLUQRI-UHFFFAOYSA-N trioctylmethylammonium bis(trifluoromethylsulfonyl)imide Chemical compound FC(F)(F)S(=O)(=O)[N-]S(=O)(=O)C(F)(F)F.CCCCCCCC[N+](C)(CCCCCCCC)CCCCCCCC LAGQNGWYNLUQRI-UHFFFAOYSA-N 0.000 claims description 2
- KIWBPDUYBMNFTB-UHFFFAOYSA-N Ethyl hydrogen sulfate Chemical compound CCOS(O)(=O)=O KIWBPDUYBMNFTB-UHFFFAOYSA-N 0.000 claims 1
- IUNCEDRRUNZACO-UHFFFAOYSA-N butyl(trimethyl)azanium Chemical compound CCCC[N+](C)(C)C IUNCEDRRUNZACO-UHFFFAOYSA-N 0.000 claims 1
- 125000004170 methylsulfonyl group Chemical group [H]C([H])([H])S(*)(=O)=O 0.000 claims 1
- 238000002425 crystallisation Methods 0.000 abstract description 20
- 230000008025 crystallization Effects 0.000 abstract description 18
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- 229910052737 gold Inorganic materials 0.000 description 35
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- AQHDANHUMGXSJZ-UHFFFAOYSA-N baicalin Natural products OC1C(O)C(C(O)CO)OC1OC(C(=C1O)O)=CC2=C1C(=O)C=C(C=1C=CC=CC=1)O2 AQHDANHUMGXSJZ-UHFFFAOYSA-N 0.000 description 22
- IKIIZLYTISPENI-ZFORQUDYSA-N baicalin Chemical compound O1[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1OC(C(=C1O)O)=CC2=C1C(=O)C=C(C=1C=CC=CC=1)O2 IKIIZLYTISPENI-ZFORQUDYSA-N 0.000 description 22
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- 230000001965 increasing effect Effects 0.000 description 4
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- JMGZEFIQIZZSBH-UHFFFAOYSA-N Bioquercetin Natural products CC1OC(OCC(O)C2OC(OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5)C(O)C2O)C(O)C(O)C1O JMGZEFIQIZZSBH-UHFFFAOYSA-N 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 3
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- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 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 3
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- IQQRAVYLUAZUGX-UHFFFAOYSA-N 1-butyl-3-methylimidazolium Chemical compound CCCCN1C=C[N+](C)=C1 IQQRAVYLUAZUGX-UHFFFAOYSA-N 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 2
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- LDVVBLGHGCHZBJ-UHFFFAOYSA-N 1-decyl-3-methylimidazolium Chemical compound CCCCCCCCCCN1C=C[N+](C)=C1 LDVVBLGHGCHZBJ-UHFFFAOYSA-N 0.000 description 1
- SXAMGRAIZSSWIH-UHFFFAOYSA-N 2-[3-[2-(2,3-dihydro-1H-inden-2-ylamino)pyrimidin-5-yl]-1,2,4-oxadiazol-5-yl]-1-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethanone Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C1=NOC(=N1)CC(=O)N1CC2=C(CC1)NN=N2 SXAMGRAIZSSWIH-UHFFFAOYSA-N 0.000 description 1
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 description 1
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- 125000003118 aryl group Chemical group 0.000 description 1
- XSGKJXQWZSFJEJ-UHFFFAOYSA-N bis(trifluoromethylsulfonyl)azanide;butyl(trimethyl)azanium Chemical compound CCCC[N+](C)(C)C.FC(F)(F)S(=O)(=O)[N-]S(=O)(=O)C(F)(F)F XSGKJXQWZSFJEJ-UHFFFAOYSA-N 0.000 description 1
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
- C07H17/06—Benzopyran radicals
- C07H17/065—Benzo[b]pyrans
- C07H17/07—Benzo[b]pyran-4-ones
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
- C07H1/08—Separation; Purification from natural products
-
- 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
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Abstract
Description
본 발명은 이온성 액체를 이용하여 천연물로부터 고순도의 플라보노이드를 효율적으로 추출 및 분리하는 방법에 관한 것이다. 구체적으로, 상기 추출 및 분리 방법은 천연물로부터 이온성 액체를 이용하여 플라보노이드를 추출하는 단계; 상기 플라보노이드 추출용액에 반용매 (anti-solvent)를 첨가하여 플라보노이드의 용석 결정화를 유도하고 1차 침전물을 얻는 단계; 상기 1차 침전물에 재결정용매 (recrystallization solvent)를 첨가하여 플라보노이드의 재결정화를 유도하고 2차 침전물을 얻는 단계; 및 상기 2차 침전물의 여과를 통해 플라보노이드를 분리하는 단계를 포함한다.The present invention relates to a method for efficiently extracting and separating high-purity flavonoids from natural products using an ionic liquid. Specifically, the extraction and separation method includes extracting flavonoids from natural products using an ionic liquid; adding an anti-solvent to the flavonoid extraction solution to induce soluble crystallization of flavonoids and to obtain a primary precipitate; adding a recrystallization solvent to the primary precipitate to induce recrystallization of flavonoids and to obtain a secondary precipitate; and separating flavonoids through filtration of the secondary precipitate.
최근 건강수명의 중요성이 강조되고 고령화 사회로 진입하면서 합성 화학물질 기반의 의약품, 건강기능식품 및 화장품보다는 천연물질 기반의 것들을 선호하는 추세이다. 이러한 추세에 더불어, 식물 추출물이 제공하는 건강상의 이점으로 인한 수요 증가와 식물 추출물의 효과 및 효능에 대한 활발한 연구가 바탕이 되어 시장 성장이 더욱 촉진될 것으로 보인다. 특히, 식물에서 발견되는 플라보노이드 (flavonoid)는 항산화, 항염증, 항균 및 항암효과로 인해서 큰 주목을 받고 있어 국내외 시장의 성장이 예측된다.Recently, as the importance of a healthy lifespan is emphasized and entering an aging society, there is a tendency to prefer natural substance-based products rather than synthetic chemical substance-based medicines, health functional foods, and cosmetics. In addition to this trend, the growing demand due to the health benefits provided by plant extracts and active research on the effects and efficacy of plant extracts are expected to further stimulate the market growth. In particular, flavonoids found in plants are attracting great attention due to their antioxidant, anti-inflammatory, antibacterial and anticancer effects, and the growth of domestic and foreign markets is expected.
일반적으로 식물 등과 같은 천연물로부터 플라보노이드와 같은 활성성분을 추출 및 분리하기 위한 과정에서 유기용매를 사용한다. 그러나 유기용매를 이용하여 천연물에서 원하는 활성성분을 추출하고 분리해내는 작업은 매우 복잡하고 까다로워 숙련된 기술과 많은 시간을 요구한다. 용해도가 낮은 물질의 경우에는 수율 또한 낮기 때문에 많은 양의 활성성분을 얻어내기 위해서는 과량의 천연물과 유기용매가 필요하며, 결국 생산 비용이 높아진다.In general, an organic solvent is used in a process for extracting and separating active ingredients such as flavonoids from natural products such as plants. However, the extraction and separation of desired active ingredients from natural products using organic solvents is very complex and demanding, requiring skilled techniques and a lot of time. In the case of materials with low solubility, the yield is also low, so an excessive amount of natural products and organic solvents are required to obtain a large amount of active ingredients, resulting in high production costs.
기존 추출 방법의 이러한 어려움을 극복하기 위해서 다양한 연구가 진행되었다. 미국특허 2006/0099690 A1에서 천연물의 추출 단계에서 나린지네이즈 (naringinase), 글루코시데이즈 (glucosidase)와 같은 효소를 추출용매에 추가하여 플라보노이드 (flavonoid)의 추출을 강화하고, 결정화를 통해 활성성분을 얻어내는 추출 방법을 개시하고 있다. 또한, 미국특허 7595080 B2에서 열수추출을 통해 천연물에서 활성성분을 추출하고, 여과하여 얻어낸 추출액에 효소를 추가함으로써 고농도의 활성성분을 얻어내는 추출 방법을 개시하고 있다. 그러나 상기와 같이 천연물의 추출 단계에서 효소를 적용하는 방법은 특정 활성성분의 추출을 강화시킬 수 있지만, 해당 활성성분이 효소에 의해 분해되어 수율이 떨어질 수 있는 단점이 존재한다.In order to overcome these difficulties of the existing extraction method, various studies have been conducted. In U.S. Patent 2006/0099690 A1, enzymes such as naringinase and glucosidase are added to the extraction solvent in the extraction step of natural products to enhance the extraction of flavonoids, and to extract active ingredients through crystallization. The extraction method obtained is disclosed. In addition, U.S. Patent No. 7595080 B2 discloses an extraction method in which an active ingredient is extracted from a natural product through hot water extraction and an enzyme is added to an extract obtained by filtering to obtain a high concentration of active ingredient. However, the method of applying an enzyme in the extraction step of a natural product as described above can enhance the extraction of a specific active ingredient, but has a disadvantage that the yield may decrease because the active ingredient is decomposed by the enzyme.
따라서, 유기용매를 이용하여 천연물로부터 활성성분을 추출 및 분리하는 방법의 단점을 극복하기 위한 빠르고 효율적인 추출 및 분리 기술의 개발이 필요하다.Therefore, it is necessary to develop a fast and efficient extraction and separation technology to overcome the disadvantages of methods for extracting and separating active ingredients from natural products using organic solvents.
본 발명에서는 기존 추출용매인 유기용매를 대체한 새로운 소재로 이온성 액체 (Ionic liquids)를 선택하였으나, 증기압이 거의 없는 이온성 액체의 독특한 특성으로 이를 결정화 용매로 사용하거나 결정화 방법을 적용하기에 어려움이 많이 따른다.In the present invention, ionic liquids were selected as a new material that replaced organic solvents, which are existing extraction solvents, but it is difficult to use them as crystallization solvents or apply crystallization methods due to the unique characteristics of ionic liquids with almost no vapor pressure. This follows a lot.
본 발명자들은 상기의 문제점들을 해결하고자, 이온성 액체를 적용하여 천연물에서 활성성분을 고효율 및 고순도로 분리하는 방법을 개발하였다. 즉, 이온성 액체를 추출용매로 적용하여 천연물 유래 플라보노이드의 추출률을 높이고, 용석 결정화 (drowning-out crystallization) 및 재결정화를 통한 추출액의 결정화로 보다 효율적으로 높은 순도의 플라보노이드를 얻어냄으로써 본 발명을 완성하였다.In order to solve the above problems, the inventors of the present invention developed a method for separating active ingredients from natural products with high efficiency and high purity by applying an ionic liquid. That is, the present invention is completed by increasing the extraction rate of natural product-derived flavonoids by applying an ionic liquid as an extraction solvent, and obtaining high-purity flavonoids more efficiently through crystallization of the extract through falling-out crystallization and recrystallization. did
본 발명은 이온성 액체를 이용하여 천연물로부터 플라보노이드를 추출 및 분리하는 방법을 제공하는 것을 목적으로 한다.An object of the present invention is to provide a method for extracting and separating flavonoids from natural products using an ionic liquid.
또한, 본 발명은 상기 추출 방법에서 이온성 액체를 추출용매로 사용하여 천연물로부터 플라보노이드를 고수율로 추출하는 방법을 제공하는 것을 목적으로 한다.In addition, an object of the present invention is to provide a method for extracting flavonoids from natural products in high yield by using an ionic liquid as an extraction solvent in the extraction method.
또한, 본 발명은 상기 분리 방법에서 용석 결정화 및 재결정화를 적용하여 고순도의 플라보노이드를 분리하는 방법을 제공하는 것을 목적으로 한다.In addition, an object of the present invention is to provide a method for separating high-purity flavonoids by applying laxative crystallization and recrystallization in the above separation method.
본 발명은 이온성 액체를 이용하여 천연물로부터 플라보노이드를 추출 및 분리하는 방법을 제공한다.The present invention provides a method for extracting and separating flavonoids from natural products using an ionic liquid.
본 발명은the present invention
(a) 천연물로부터 이온성 액체를 이용하여 플라보노이드를 추출하는 단계;(a) extracting flavonoids from natural products using an ionic liquid;
(b) 상기 플라보노이드 추출용액에 반용매 (anti-solvent)를 첨가하여 플라보노이드의 용석 결정화를 유도하고 1차 침전물을 얻는 단계;(b) adding an anti-solvent to the flavonoid extract solution to induce soluble crystallization of the flavonoids and obtain a primary precipitate;
(c) 상기 1차 침전물에 재결정용매 (recrystallization solvent)를 첨가하여 플라보노이드의 재결정화를 유도하고 2차 침전물을 얻는 단계; 및(c) inducing recrystallization of flavonoids by adding a recrystallization solvent to the primary precipitate and obtaining a secondary precipitate; and
(d) 상기 2차 침전물의 여과를 통해 플라보노이드를 분리하는 단계를 포함한다.(d) separating flavonoids through filtration of the secondary precipitate.
본 발명의 추출 및 분리 방법을 통해 고수율 및 고순도의 플라보노이드를 효율적으로 추출 및 분리할 수 있다.The extraction and separation method of the present invention can efficiently extract and separate flavonoids in high yield and purity.
본 발명은 이온성 액체를 이용하여 천연물로부터 고순도의 플라보노이드를 추출하는 방법을 제공한다. 이를 통해 천연물에서 플라보노이드를 효율적으로 추출할 수 있어, 경제적인 측면에서 이점을 제공한다.The present invention provides a method for extracting high-purity flavonoids from natural products using an ionic liquid. Through this, it is possible to efficiently extract flavonoids from natural products, providing advantages in terms of economy.
또한, 본 발명을 통해 이온성 액체를 사용한 후 이를 제거하여 목적하는 플라보노이드를 선택적으로 얻어낼 수 있으므로, 상기 플라보노이드는 기존의 표준품을 대신하여 실험 및 산업 등에 다양하게 사용될 수 있다.In addition, since a desired flavonoid can be selectively obtained by removing the ionic liquid after using the present invention, the flavonoid can be used in various fields such as experiments and industries instead of existing standard products.
도 1은 이온성 액체 및 알코올 용매에 따른 황금으로부터 바이칼린의 수율 및 괴화로부터 루틴의 수율을 나타낸다.
도 2는 이온성 액체와 알코올 용매의 혼합 농도에 따른 황금으로부터 바이칼린의 수율을 나타낸다.
도 3은 이온성 액체와 천연물의 혼합 비율에 따른 황금으로부터 바이칼린의 수율을 나타낸다.
도 4는 초음파 추출 시간에 따른 황금으로부터 바이칼린의 수율을 나타낸다.
도 5는 초음파 추출 온도에 따른 황금으로부터 바이칼린의 수율을 나타낸다.
도 6은 재결정용매에 따른 황금으로부터 바이칼린의 수율 및 회수율을 나타낸다.
도 7은 황금으로부터 분리된 플라보노이드의 1H-NMR 분석 결과를 나타낸다.
도 8은 괴화로부터 분리된 플라보노이드의 HPLC 분석 결과를 나타낸다.Figure 1 shows the yield of baicalin from gold and the yield of rutin from agglomeration according to the ionic liquid and alcohol solvent.
Figure 2 shows the yield of baicalin from gold according to the mixed concentration of the ionic liquid and the alcohol solvent.
Figure 3 shows the yield of baicalin from gold according to the mixing ratio of the ionic liquid and the natural product.
Figure 4 shows the yield of baicalin from gold according to the ultrasonic extraction time.
Figure 5 shows the yield of baicalin from gold according to the ultrasonic extraction temperature.
Figure 6 shows the yield and recovery of baicalin from gold according to the recrystallization solvent.
Figure 7 shows the results of 1 H-NMR analysis of flavonoids isolated from gold.
8 shows the results of HPLC analysis of flavonoids isolated from necrosis.
이하, 첨부한 도면을 참조하여 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있도록 본원의 실시태양 및 실시예를 상세히 설명한다. 그러나 본원은 여러 가지 형태로 구현될 수 있으며 여기에서 설명하는 실시태양 및 실시예에 한정되지 않는다.Hereinafter, with reference to the accompanying drawings, embodiments and examples of the present disclosure will be described in detail so that those skilled in the art can easily practice the present invention. However, the present application may be implemented in various forms and is not limited to the embodiments and examples described herein.
본원 명세서 전체에서, 어떤 부분이 어떤 구성 요소를 "포함" 한다고 할 때, 이는 특별히 반대되는 기재가 없는 한 다른 구성 요소를 제외하는 것이 아니라 다른 구성 요소를 더 포함할 수 있는 것을 의미한다.Throughout the present specification, when a part "includes" a certain component, it means that it may further include other components without excluding other components unless otherwise stated.
본 발명에 사용된 “이온성 액체 (ionic liquids)”는 양이온과 음이온이 크기의 비대칭성으로 인해 결정체를 이루지 못하고 액체 상태로 존재하는 물질을 의미하고, 낮은 휘발성, 열적 안정성, 높은 이온전도성, 넓은 전기화학적 안정성, 낮은 증기압 등의 특성을 갖는다.As used in the present invention, “ionic liquids” refer to substances in which cations and anions do not form crystals due to size asymmetry and exist in a liquid state, and have low volatility, thermal stability, high ionic conductivity, wide It has characteristics such as electrochemical stability and low vapor pressure.
본 발명에 사용된 “플라보노이드 (flavonoid)”는 15개의 탄소로 이루어진 3개 고리의 골격을 가지며 평면의 방향족 고리 구조를 가지는 물질을 의미한다. 식물에서 발견되는 플라보노이드는 배당체나 비당체로 존재하며, 인간에서 각기 다른 생체이용률을 가진다.As used herein, “flavonoid” refers to a substance having a planar aromatic ring structure with a three-ring backbone of 15 carbon atoms. Flavonoids found in plants exist as glycosides or non-glycosides, and each has a different bioavailability in humans.
본 발명에 사용된 “결정화 (crystallization)”는 분자인식과 자기조립의 원리를 통해 용액에서 용질을 결정으로 석출시키는 고-액분리기술을 의미한다. 유기용매를 이용한 분획이나 컬럼 크로마토그래피에 비해 더 쉽고 효율적으로 활성성분을 분리해 낼 수 있는 장점이 있으며, 분리된 성분의 품질이 우수하고 수득률 또한 높다. 결정화 방법으로는 냉각 결정화 (cooling crystallization), 증발 결정화 (evaporative crystallization), 침전 (precipitation) 및 용석 결정화 (drowning-out crystallization) 등이 포함된다.“Crystallization” used in the present invention refers to a solid-liquid separation technique in which a solute is precipitated as crystals from a solution through the principle of molecular recognition and self-assembly. Compared to fractionation or column chromatography using an organic solvent, it has the advantage of being able to separate the active ingredient more easily and efficiently, and the quality of the separated ingredient is excellent and the yield is also high. Crystallization methods include cooling crystallization, evaporative crystallization, precipitation, and drowning-out crystallization.
본 발명은 이온성 액체를 이용하여 천연물로부터 고순도의 플라보노이드를 효율적으로 추출 및 분리하는 방법에 관한 것으로,The present invention relates to a method for efficiently extracting and separating high-purity flavonoids from natural products using an ionic liquid.
(a) 천연물에 이온성 액체를 첨가하여 플라보노이드 추출용액을 수득하는 단계;(a) obtaining a flavonoid extract solution by adding an ionic liquid to a natural product;
(b) 상기 플라보노이드 추출용액에 반용매 (anti-solvent)를 첨가하여 1차 침전물을 수득하는 단계;(b) obtaining a primary precipitate by adding an anti-solvent to the flavonoid extract solution;
(c) 상기 1차 침전물에 재결정용매 (recrystallization solvent)를 첨가하여 2차 침전물을 수득하는 단계; 및(c) obtaining a secondary precipitate by adding a recrystallization solvent to the primary precipitate; and
(d) 상기 2차 침전물을 여과하는 단계를 포함한다.(d) filtering the secondary precipitate.
본 발명의 추출 대상이 되는 천연물은 꿀풀과 (Labiatae)에 속하는 식물인 속썩은풀 (Scutellaria baicalensis Georg)의 뿌리 또는 콩과 (Leguminosae)에 속하는 식물인 회화나무 (Sophora japonica)의 미개화일 수 있다.The natural product to be extracted in the present invention may be the root of Scutellaria baicalensis Georg, a plant belonging to the Lamiaceae family (Labiatae), or the unflowered Sophora japonica , a plant belonging to the Leguminosae family.
일 실시태양에서, 상기 천연물의 부위는 속썩은풀의 뿌리로서 그대로 또는 주피를 제거한 것, 또는 회화나무의 미개화일 수 있으며, 구체적으로 황금 (The roots of Scutellaria baicalensis) 또는 괴화 (The flowers of Sophora japonica)일 수 있다.In one embodiment, the part of the natural product may be the root of the rotten grass intact or with the perianth removed, or the unflowered tree of the fig tree, specifically the roots of Scutellaria baicalensis or the flowers of Sophora japonica ) can be.
본 발명의 플라보노이드는 바이칼린 또는 루틴일 수 있다.A flavonoid of the present invention may be baicalin or rutin.
상기 이온성 액체를 구성하는 양이온은 몰포리늄, 이미다졸리움 양이온, 옥사졸리움 양이온, 피페리디늄 양이온, 피라지니움 양이온, 피라졸리움 양이온, 피리다지니늄 양이온, 피리디늄 양이온, 피리미디늄 양이온, 피롤리디늄 양이온, 피롤리늄 양이온, 피롤리움 양이온, 설포늄 양이온, 포스포늄 양이온 및 트리아졸륨 양이온으로 구성된 군으로부터 선택될 수 있고, 바람직하게는 이미다졸리움 및 피리디늄 양이온으로 구성된 군으로부터 선택될 수 있으며, 더욱 바람직하게는 피리디늄 양이온일 수 있다.The cations constituting the ionic liquid are morpholinium, imidazolium cation, oxazolium cation, piperidinium cation, pyrazinium cation, pyrazolium cation, pyridazininium cation, pyridinium cation, pyrimidinium cation , It may be selected from the group consisting of pyrrolidinium cations, pyrrolinium cations, pyrroleium cations, sulfonium cations, phosphonium cations and triazolium cations, preferably selected from the group consisting of imidazolium and pyridinium cations It may be, and more preferably may be a pyridinium cation.
상기 이온성 액체를 구성하는 음이온은 에틸설페이트 음이온, 토실레이트 음이온, 메탄설폰 음이온, 테트라플루오로보레이트 음이온, 헥사플루오르포스페이트 음이온, 헥사플루오르안티모네이트 음이온 및 테트라클로로알루미네이트, 브로마이드 음이온 및 클로라이드 음이온으로 구성된 군으로부터 선택될 수 있고, 바람직하게는 테트라플루오로보레이트 음이온, 브로마이드 음이온 및 클로라이드로 음이온으로 구성된 군으로부터 선택될 수 있으며, 더욱 바람직하게는 브로마이드 음이온일 수 있다.Anions constituting the ionic liquid include ethylsulfate anion, tosylate anion, methanesulfone anion, tetrafluoroborate anion, hexafluorophosphate anion, hexafluoroantimonate anion, tetrachloroaluminate, bromide anion and chloride anion. It may be selected from the group consisting of, preferably selected from the group consisting of tetrafluoroborate anion, bromide anion and chloride anion, more preferably bromide anion.
상기 이온성 액체는 1-부틸-3-메틸이미다졸리움 브로마이드, 1-부틸-3-메틸이미다졸리움 클로라이드, 1-부틸-3-메틸이미다졸리움 테트라플루오로보레이트, 1-부틸-3-메틸이미다졸리움 헥사플루오로포스페이트, 1-메틸-3-에틸이미다졸리움 브로마이드, 1-옥틸-3-메틸이미다졸리움 브로마이드, 1-에틸피리디늄 브로마이드, 1-에틸-3-메틸이미다졸리움 에틸설페이트, 1-에틸-3-메틸이미다졸리움 테트라플루오로보레이트, 1-에틸-3-메틸이미다졸리움 토실레이트, 1-에틸-3-메틸이미다졸리움 메탄설포네이트, 1-에틸-3-메틸이미다졸리움 프리플레이트, 1-에틸-3-메틸이미다졸리움 디시안아미드, 1-부틸-3-메틸이미다졸리움 테트라플루오르보레이트, 1-부틸-3-메틸이미다졸리움 비스(트리플푸오로메틸설포닐)이미드, 1-부틸-3-메틸이미다졸리움 디시안아미드, 1-부틸-3-메틸이미다졸리움 트리플레이트, 1-부틸-3-메틸이미다졸리움 메탄설포네이트, 1,3-디데실-2-메틸이미다졸리움 비스(트리플푸오로메틸설포닐)이미드, 1,3-디알릴이미다졸리움 테트라플르오르보레이트, 1-부틸-2,3-디메틸이미다졸리움 트리플레이트, 1-부틸-2,3-디메틸이미다졸리움 테트라플루오로보레이트, 1-(2-히드록시에틸)-3-메틸이미다졸리움 테트라플루오로보레이트, 1-데실-3-메틸이미다졸리움 테트라플루오로보레이트, 1-부틸-3-메틸피리디늄 테트라플루오로보레이트, 1-부틸-4-메틸피리디늄 테트라플루오로보레이트, 1-알릴-3-에틸이미다졸리움 테트라플루오로보레이트, 1-메틸-1프로필피페리디늄 비스(트리플루오로메틸설포닐)이미드, 메틸트리옥틸암모늄 비스(트리플루오로메틸설포닐)이미드, 부틸트리메틸암모늄 비스(트리플루오로메틸설포닐)이미드, 트리에틸설포늄 비스(트리플루오로메틸설포닐)이미드 및 디에틸메틸설포늄 비스(트리플루오로메틸설포닐)이미드로 구성된 군으로부터 선택될 수 있고, 바람직하게는 1-에틸-3-메틸이미다졸리움 테트라플루오로보레이트, 1,3-디알릴이미다졸리움 테트라플르오르보레이트, 1-부틸-2,3-디메틸이미다졸리움 테트라플루오로보레이트, 1-(2-히드록시에틸)-3-메틸이미다졸리움 테트라플루오로보레이트, 1-데실-3-메틸이미다졸리움 테트라플루오로보레이트, 1-부틸-3-메틸피리디늄 테트라플루오로보레이트, 1-부틸-4-메틸피리디늄 테트라플루오로보레이트 또는 1-알릴-3-에틸이미다졸리움 테트라플루오로보레이트 군으로부터 선택되고, 바람직하게는 1-부틸-3-메틸이미다졸리움 브로마이드, 1-부틸-3-메틸이미다졸리움 클로라이드, 1-부틸-3-메틸이미다졸리움 테트라플루오로보레이트, 1-부틸-3-메틸이미다졸리움 헥사플루오로포스페이트, 1-메틸-3-에틸이미다졸리움 브로마이드, 1-옥틸-3-메틸이미다졸리움 브로마이드, 1-에틸피리디늄 브로마이드 군으로부터 선택될 수 있으며, 더욱 바람직하게는 1-에틸피리디늄 브로마이드일 수 있다.The ionic liquid is 1-butyl-3-methylimidazolium bromide, 1-butyl-3-methylimidazolium chloride, 1-butyl-3-methylimidazolium tetrafluoroborate, 1-butyl-3- Methylimidazolium hexafluorophosphate, 1-methyl-3-ethylimidazolium bromide, 1-octyl-3-methylimidazolium bromide, 1-ethylpyridinium bromide, 1-ethyl-3-methylimidazolium Ethyl sulfate, 1-ethyl-3-methylimidazolium tetrafluoroborate, 1-ethyl-3-methylimidazolium tosylate, 1-ethyl-3-methylimidazolium methanesulfonate, 1-ethyl-3 -Methylimidazolium preplate, 1-ethyl-3-methylimidazolium dicyanamide, 1-butyl-3-methylimidazolium tetrafluoroborate, 1-butyl-3-methylimidazolium bis(triple fu) Oromethylsulfonyl) imide, 1-butyl-3-methylimidazolium dicyanamide, 1-butyl-3-methylimidazolium triflate, 1-butyl-3-methylimidazolium methanesulfonate, 1 ,3-Didecyl-2-methylimidazolium bis(triplefluoromethylsulfonyl)imide, 1,3-diallylimidazolium tetrafluoroborate, 1-butyl-2,3-dimethylimidazolium Triflate, 1-butyl-2,3-dimethylimidazolium tetrafluoroborate, 1-(2-hydroxyethyl)-3-methylimidazolium tetrafluoroborate, 1-decyl-3-methylimida Zolium tetrafluoroborate, 1-butyl-3-methylpyridinium tetrafluoroborate, 1-butyl-4-methylpyridinium tetrafluoroborate, 1-allyl-3-ethylimidazolium tetrafluoroborate, 1 -Methyl-1propylpiperidinium bis(trifluoromethylsulfonyl)imide, methyltrioctylammonium bis(trifluoromethylsulfonyl)imide, butyltrimethylammonium bis(trifluoromethylsulfonyl)imide , triethylsulfonium bis(trifluoromethylsulfonyl)imide and diethylmethylsulfonium bis(trifluoromethylsulfonyl)imide, preferably 1-ethyl-3- Methylimidazolium tetrafluoroborate, 1,3-diallylimidazolium tetrafluoroborate, 1-butyl-2,3-dimethylimidazolium tetrafluoro Borate, 1-(2-hydroxyethyl)-3-methylimidazolium tetrafluoroborate, 1-decyl-3-methylimidazolium tetrafluoroborate, 1-butyl-3-methylpyridinium tetrafluoro selected from the group of borates, 1-butyl-4-methylpyridinium tetrafluoroborate or 1-allyl-3-ethylimidazolium tetrafluoroborate, preferably 1-butyl-3-methylimidazolium bromide; 1-butyl-3-methylimidazolium chloride, 1-butyl-3-methylimidazolium tetrafluoroborate, 1-butyl-3-methylimidazolium hexafluorophosphate, 1-methyl-3-ethyl It may be selected from the group of midazolium bromide, 1-octyl-3-methylimidazolium bromide, and 1-ethylpyridinium bromide, more preferably 1-ethylpyridinium bromide.
구체적으로, 상기 (a) 단계는 건조된 천연물을 분쇄하고 이온성 액체를 추출용매로 사용하여 초음파 추출한 후, 천연물-이온성액체의 혼합물에서 고형분을 제거하여 플라보노이드 추출용액을 얻어내는 과정을 포함한다.Specifically, the step (a) includes a process of pulverizing the dried natural product, performing ultrasonic extraction using an ionic liquid as an extraction solvent, and removing solids from the natural product-ionic liquid mixture to obtain a flavonoid extract solution. .
상기 (a) 단계에서, 천연물과 이온성 액체는 1:3 내지 1:30의 중량비로 포함될 수 있으며, 바람직하게는 1:10일 수 있다.In step (a), the natural product and the ionic liquid may be included in a weight ratio of 1:3 to 1:30, preferably 1:10.
상기 (a) 단계에서, 이온성 액체는 알코올 용매와 혼합하여 추출용매로 사용될 수 있다. 상기 알코올 용매는 저급 알코올 또는 알코올 수용액일 수 있다. 또한, 알코올 용매에 혼합되는 이온성 액체의 농도는 6M 이하일 수 있으며, 바람직하게는 5M 이하일 수 있으며, 더욱 바람직하게는 1 내지 4M일 수 있다.In step (a), the ionic liquid may be used as an extraction solvent by mixing with an alcohol solvent. The alcohol solvent may be a lower alcohol or an aqueous alcohol solution. In addition, the concentration of the ionic liquid mixed with the alcohol solvent may be 6M or less, preferably 5M or less, and more preferably 1 to 4M.
상기 (a) 단계에서, 초음파 추출 온도는 0 내지 60°C일 수 있고, 바람직하게는 30 내지 50°C일 수 있다.In the step (a), the ultrasonic extraction temperature may be 0 to 60 °C, preferably 30 to 50 °C.
상기 (a) 단계에서, 초음파 추출 시간은 24시간 이하일 수 있고, 바람직하게는 120분 이하일 수 있고, 더욱 바람직하게는 90분일 수 있다.In the step (a), the ultrasonic extraction time may be 24 hours or less, preferably 120 minutes or less, and more preferably 90 minutes.
상기 (a) 단계에서, 고형분의 제거에는 원심분리 또는 여과를 적용할 수 있다.In the step (a), centrifugation or filtration may be applied to remove the solid content.
구체적으로, 상기 (b) 단계는 플라보노이드 추출용액에 무기 또는 유기용매를 반용매 (anti-solvent)로 첨가하여 용석 결정화 (drowning-out crystallization)를 유도한 후, 용매를 제거한 1차 침전물을 얻어내는 과정을 포함한다.Specifically, in the step (b), an inorganic or organic solvent is added as an anti-solvent to the flavonoid extract solution to induce drowning-out crystallization, and then the solvent is removed to obtain a primary precipitate. include the process
상기 반용매 (anti-solvent)는 극성도 표에서 메탄올 이하의 극성도를 가지며 추출액과 혼합되는 용매로 구성된 군으로부터 선택될 수 있고, 바람직하게는 메틸아세테이트, 메틸렌클로라이드, 벤젠, 부탄올, 아세토니트릴, 아세톤, 에탄올, 에틸아세테이트, 이소프로필알콜, 클로로포름으로 구성된 군에서 선택될 수 있으며, 더욱 바람직하게는 아세토니트릴일 수 있다.The anti-solvent has a polarity of less than methanol in the polarity table and may be selected from the group consisting of solvents that are mixed with the extract, preferably methyl acetate, methylene chloride, benzene, butanol, acetonitrile, It may be selected from the group consisting of acetone, ethanol, ethyl acetate, isopropyl alcohol, and chloroform, more preferably acetonitrile.
상기 (b) 단계에서, 플라보노이드 추출용액과 반용매가 1:3 내지 1:30의 부피비로 포함될 수 있고, 바람직하게는 1:5 내지 1:10의 부피비로 포함될 수 있다.In the step (b), the flavonoid extract solution and the antisolvent may be included in a volume ratio of 1:3 to 1:30, preferably 1:5 to 1:10.
상기 (b) 단계에서, 용석 결정화 유도 시간은 24시간일 수 있고, 바람직하게는 60분 이하일 수 있다.In the step (b), the time required to induce molten metal crystallization may be 24 hours, preferably 60 minutes or less.
상기 (b) 단계에서, 용매의 제거에는 원심분리를 적용할 수 있다.In step (b), centrifugation may be applied to remove the solvent.
구체적으로, 상기 (c) 단계는 1차 침전물에 무기 또는 유기용매를 재결정용매 (recrystallization solvent)로 첨가하여 플라보노이드의 재결정화를 유도한 후, 2차 침전물을 얻어내는 과정을 포함한다.Specifically, the step (c) includes adding an inorganic or organic solvent as a recrystallization solvent to the primary precipitate to induce recrystallization of flavonoids, and then obtaining secondary precipitates.
상기 재결정용매 (recrystallization solvent)는 상기 1차 침전물을 녹일 수 있을 정도의 극성을 가지는 용매로 구성된 군으로부터 선택될 수 있고, 바람직하게는 물, 메탄올, 아세트산, 에탄올, 포름산으로 구성된 군으로부터 선택되는 무기 또는 유기용매의 pH를 조절한 것일 수 있으며, 더욱 바람직하게는 0.1% 수용성 포름산일 수 있다.The recrystallization solvent may be selected from the group consisting of solvents having a degree of polarity capable of dissolving the primary precipitate, preferably inorganic selected from the group consisting of water, methanol, acetic acid, ethanol, and formic acid Alternatively, the pH of the organic solvent may be adjusted, and more preferably 0.1% water-soluble formic acid.
상기 (c) 단계에서, 1차 침전물과 재결정용매가 1:3 내지 1:30의 비로 포함될 수 있고, 가장 바람직하게는 1:5 내지 1:10의 비로 포함될 수 있다.In step (c), the primary precipitate and the recrystallization solvent may be included in a ratio of 1:3 to 1:30, and most preferably in a ratio of 1:5 to 1:10.
상기 (c) 단계에서, 반응 시간은 24시간일 수 있고, 바람직하게는 4 내지 24시간일 수 있다.In step (c), the reaction time may be 24 hours, preferably 4 to 24 hours.
구체적으로, 상기 (d) 단계는 2차 침전물을 감압여과 시킨 후, 친수성 용매로 씻어내어 비유효성분을 제거하고, 용매를 완전히 제거하여 최종 플라보노이드를 얻어내는 과정을 포함한다.Specifically, the step (d) includes a process of filtering the secondary precipitate under reduced pressure, washing it with a hydrophilic solvent to remove non-active ingredients, and completely removing the solvent to obtain the final flavonoid.
상기 친수성 용매는 물, 메탄올, 에탄올에서 선택되는 무기 또는 유기용매일 수 있고, 바람직하게는 물일 수 있다.The hydrophilic solvent may be an inorganic or organic solvent selected from water, methanol and ethanol, preferably water.
상기 (d) 단계에서, 용매의 제거에는 오븐건조법을 적용할 수 있다.In the step (d), an oven drying method may be applied to remove the solvent.
본 발명에 사용된 "추출물"은 당업계에서 조추출물 (crude extract)로 통용되는 의미를 갖지만, 광의적으로 추출물을 추가적으로 분획 (fractionation)한 분획물도 포함한다. 즉 추출물은 추출용매를 이용하여 얻은 것뿐만 아니라, 여기에 정제 과정을 추가적으로 적용하여 얻은 것을 포함한다. 예컨대, 상기 추출물을 일정한 분자량 컷-오프 값을 갖는 한외여과막을 통과시켜 얻은 분획, 다양한 크로마토그래프 (크기, 전하, 소수성 또는 친화성에 다른 분리를 위해 제작된 것)에 의한 분리 등, 추가적으로 실시된 다양한 정제 방법을 통해 얻어진 분획도 본 발명의 추출물에 포함된다. 또한 상기 추출물은 용액 (유동엑스), 농축물 (연조엑스) 또는 건조 분말상태일 수 있으며, 이에 제한되지 않는다.The term "extract" used in the present invention has a meaning commonly used in the art as a crude extract, but also includes a fraction obtained by further fractionating the extract in a broad sense. That is, the extract includes not only those obtained by using an extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, separation by various chromatographs (designed for separation different in size, charge, hydrophobicity or affinity), etc. Fractions obtained through the purification method are also included in the extract of the present invention. In addition, the extract may be in the form of a solution (liquid extract), concentrate (softened extract) or dry powder, but is not limited thereto.
이하 실시예를 통하여 본 발명을 더욱 상세하게 설명하고자 하나, 하기의 실시예는 단지 설명의 목적을 위한 것이며 본원 발명의 범위를 한정하고자 하는 것은 아니다.The present invention will be described in more detail through the following examples, but the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.
[실시예 1] [Example 1]
생약으로부터 플라보노이드를 추출하기 위한 이온성 액체Ionic Liquids for Extraction of Flavonoids from Herbal Medicines
추출 생약으로 황금 (The roots of Scutellaria baicalensis) 및 괴화 (The flowers of Sophora japonica)를 선정하고, 플라보노이드를 추출하기 위한 추출용매를 확인하기 위하여 하기 실험을 수행하였다.Gold as an extract herbal medicine (The roots of Scutellaria baicalensis ) and necrosis (The flowers of Sophora japonica ) were selected, and the following experiment was performed to confirm an extraction solvent for extracting flavonoids.
구체적으로, 메탄올, 70% 메탄올 수용액, 70% 에탄올 수용액에 1-부틸-3-메틸이미다졸리움 브로마이드 ([Bmim]Br), 1-부틸-3-메틸이미다졸리움 클로라이드 ([Bmim]Cl), 1-부틸-3-메틸이미다졸리움 테트라플루오로보레이트 ([Bmim]BF4), 1-에틸-3-메틸이미다졸리움 브로마이드 ([Emim]Br), 1-옥틸-3-메틸이미다졸리움 브로마이드 ([Omim]Br), 1-에틸피리디늄 브로마이드 ([EtPyr]Br)가 각각 혼합된 18가지 이온성 액체를 사용하여 황금으로부터 추출된 바이칼린의 수율 및 괴화로부터 추출된 루틴의 수율을 측정하였고, 그 결과를 하기 도 1에 나타내었다.Specifically, 1-butyl-3-methylimidazolium bromide ([Bmim]Br), 1-butyl-3-methylimidazolium chloride ([Bmim]Cl) in methanol, 70% methanol aqueous solution, and 70% ethanol aqueous solution , 1-butyl-3-methylimidazolium tetrafluoroborate ([Bmim]BF4), 1-ethyl-3-methylimidazolium bromide ([Emim]Br), 1-octyl-3-methylimidazolium Yields of baicalin extracted from gold and rutin extracted from agglomerate were measured using 18 ionic liquids mixed with bromide ([Omim]Br) and 1-ethylpyridinium bromide ([EtPyr]Br), respectively. And the results are shown in Figure 1 below.
도 1에 나타낸 바와 같이, 상기 18가지의 이온성 액체를 추출용매로 사용하여 바이칼린의 수율을 측정한 결과, 1-부틸-3-메틸이미다졸리움 브로마이드, 1-부틸-3-메틸이미다졸리움 클로라이드, 1-에틸-3-메틸이미다졸리움 브로마이드, 또는 1-에틸피리디늄 브로마이드가 혼합된 이온성 액체에서 높은 수율을 나타내었고, 특히 황금의 경우 1-에틸피리디늄 브로마이드가 결정화 단계에서 가장 적합한 용매로 선택되었다. 괴화의 경우 1-에틸-3-메틸이미다졸리움 브로마이드가 결정화 단계에서 가장 적합한 용매로 선택되었다.As shown in FIG. 1, as a result of measuring the yield of baicalin using the 18 ionic liquids as extraction solvents, 1-butyl-3-methylimidazolium bromide and 1-butyl-3-methylimida The ionic liquid in which zolium chloride, 1-ethyl-3-methylimidazolium bromide, or 1-ethylpyridinium bromide were mixed showed high yields, especially in the case of gold, 1-ethylpyridinium bromide was the most dominant in the crystallization step. A suitable solvent was selected. For agglomeration, 1-ethyl-3-methylimidazolium bromide was chosen as the most suitable solvent for the crystallization step.
또한, 황금의 경우 70% 에탄올 수용액과 혼합된 이온성 액체를 사용하였을 경우 전체적으로 더 높은 수율을 나타내었고, 괴화의 경우 70% 메탄올 수용액을 이온성 액체와 혼합하여 사용하였을 경우 전체적으로 더 높은 수율을 나타내었다.In addition, in the case of gold, when the ionic liquid mixed with 70% ethanol aqueous solution was used, the overall yield was higher, and in the case of blackening, the overall yield was higher when 70% methanol aqueous solution was mixed with the ionic liquid. was
하기의 실시예에서는 황금을 선택하여 바이칼린의 추출 조건 최적화 과정을 진행하였다.In the following examples, gold was selected to proceed with the process of optimizing baicalin extraction conditions.
[실시예 2] [Example 2]
황금 (The roots of The roots of gold Scutellaria baicalensisScutellaria baicalensis )으로부터 플라보노이드를 추출하기 위한 이온성 액체의 농도) Concentration of ionic liquid to extract flavonoids from
황금으로부터 플라보노이드를 추출하기 위한 이온성 액체의 농도를 확인하기 위하여, 70% 에탄올 수용액에 1-에틸피리디늄 브로마이드를 1, 2, 3, 4, 5 및 6 M 농도로 혼합하여 바이칼린의 수율을 측정하였고, 그 결과를 도 2에 나타내었다. 이온성 액체 대 황금의 비율은 10:1mL/g, 추출 시간은 90분, 추출 온도는 50˚C로 고정하였다.In order to confirm the concentration of the ionic liquid for extracting flavonoids from gold, 1-ethylpyridinium bromide was mixed with 70% ethanol aqueous solution at concentrations of 1, 2, 3, 4, 5 and 6 M to determine the yield of baicalin. It was measured, and the results are shown in FIG. 2 . The ratio of ionic liquid to gold was fixed at 10:1mL/g, extraction time was 90 minutes, and extraction temperature was 50˚C.
도 2에 나타낸 바와 같이, 상기 6가지 농도로 혼합한 이온성 액체를 사용하여 바이칼린의 수율을 측정한 결과, 1M 내지 4M 농도에서 높은 수율을 나타내었고, 5M 농도 이상에서는 수율이 감소하는 것을 확인하였다.As shown in Figure 2, as a result of measuring the yield of baicalin using the ionic liquid mixed at the above six concentrations, it was confirmed that the yield was high at 1M to 4M concentration, and the yield decreased at 5M concentration or higher. did
[실시예 3] [Example 3]
황금 (The roots of The roots of gold Scutellaria baicalensisScutellaria baicalensis )으로부터 플라보노이드를 추출하기 위한 이온성 액체의 비율) Ratio of ionic liquid to extract flavonoids from
황금으로부터 플라보노이드를 추출하기 위한 이온성 액체의 비율을 확인하기 위하여, 이온성 액체 대 황금의 비율을 3:1, 10:1, 15:1, 20:1, 25:1 및 30:1로 설정하여 바이칼린의 수율을 측정하였고, 그 결과를 도 3에 나타내었다. 이온성 액체의 농도는 3.5M, 추출 시간은 90분, 추출 온도는 50˚C로 고정하였다.In order to confirm the ratio of ionic liquid to extract flavonoids from gold, the ratio of ionic liquid to gold was set at 3:1, 10:1, 15:1, 20:1, 25:1 and 30:1 The yield of baicalin was measured, and the results are shown in FIG. 3 . The concentration of the ionic liquid was fixed at 3.5M, the extraction time was 90 minutes, and the extraction temperature was 50˚C.
도 3에 나타낸 바와 같이, 이온성 액체와 황금을 상기 6가지 비율로 혼합하여 바이칼린의 수율을 측정한 결과, 10:1까지 수율이 증가하는 것을 확인하였고, 이후 수율이 점차 감소하는 것을 확인하였다.As shown in FIG. 3, as a result of measuring the yield of baicalin by mixing the ionic liquid and gold in the above six ratios, it was confirmed that the yield increased up to 10: 1, and then the yield gradually decreased. .
[실시예 4] [Example 4]
황금 (The roots of The roots of gold Scutellaria baicalensisScutellaria baicalensis )으로부터 플라보노이드를 추출하기 위한 추출 시간) Extraction time to extract flavonoids from
황금으로부터 플라보노이드를 추출하기 위한 추출 시간을 확인하기 위하여, 이온성 액체와 황금을 혼합한 후 초음파 추출 시간을 0, 30, 60, 90 및 120분으로 설정하여 바이칼린의 수율을 측정하였고, 그 결과는 도 4에 나타내었다. 이온성 액체의 농도는 3.5M, 이온성 액체 대 황금의 비율은 10:1mL/g, 추출 온도는 50˚C로 고정하였다.In order to confirm the extraction time for extracting flavonoids from gold, after mixing the ionic liquid and gold, the yield of baicalin was measured by setting the ultrasonic extraction time to 0, 30, 60, 90 and 120 minutes. is shown in Figure 4. The concentration of the ionic liquid was 3.5M, the ratio of ionic liquid to gold was 10:1mL/g, and the extraction temperature was fixed at 50˚C.
도 4에 나타낸 바와 같이, 상기 5종류의 시간에 따라 초음파 추출을 한 후 바이칼린의 수율을 측정한 결과, 90분까지 바이칼린의 수율이 증가하는 것을 확인하였고, 이후 약간 감소하는 것을 확인하였다.As shown in FIG. 4, as a result of measuring the yield of baicalin after ultrasonic extraction according to the five types of time, it was confirmed that the yield of baicalin increased until 90 minutes, and then slightly decreased.
[실시예 5] [Example 5]
황금 (The roots of The roots of gold Scutellaria baicalensisScutellaria baicalensis )으로부터 플라보노이드를 추출하기 위한 추출 온도) Extraction temperature for extracting flavonoids from
황금으로부터 플라보노이드를 추출하기 위한 추출 온도를 확인하기 위하여, 이온성 액체와 황금을 혼합한 후 초음파 추출 온도 (초음파 수조의 설정 온도)를 20, 30, 40, 50 및 60˚C로 설정하여 바이칼린의 수율을 측정하였고, 그 결과를 도 5에 나타내었다. 이온성 액체의 농도는 3.5M, 이온성 액체 대 황금의 비율은 10:1mL/g, 추출 시간은 90분으로 고정하였다.In order to confirm the extraction temperature for extracting flavonoids from gold, after mixing the ionic liquid and gold, the ultrasonic extraction temperature (set temperature of the ultrasonic bath) was set to 20, 30, 40, 50 and 60˚C to baicalin The yield of was measured, and the results are shown in FIG. 5. The concentration of the ionic liquid was fixed at 3.5M, the ratio of ionic liquid to gold was 10:1mL/g, and the extraction time was 90 minutes.
도 5에 나타낸 바와 같이, 상기 5가지 온도에 따라 초음파 추출을 한 후 바이칼린의 수율을 측정한 결과, 60˚C까지 바이칼린의 수율이 증가하는 것을 확인하였다. 그러나 고온에서 화합물이 용매에 의해 용해 또는 분해될 수 있음을 고려하여, 천연물로부터 플라보노이드를 추출하기 위한 추출 온도를 50˚C로 선택하였다.As shown in Figure 5, as a result of measuring the yield of baicalin after ultrasonic extraction according to the above five temperatures, it was confirmed that the yield of baicalin increased up to 60 °C. However, considering that compounds can be dissolved or decomposed by solvents at high temperatures, 50 °C was selected as an extraction temperature for extracting flavonoids from natural products.
[실시예 6] [Example 6]
황금 (The roots of The roots of gold Scutellaria baicalensisScutellaria baicalensis )으로부터 플라보노이드를 분리하기 위한 재결정용매) Recrystallization solvent for separating flavonoids from
황금으로부터 플라보노이드를 분리하기 위한 재결정용매를 확인하기 위하여, 하기와 같이 실험을 진행하였다.In order to confirm the recrystallization solvent for separating flavonoids from gold, the experiment was conducted as follows.
각 알코올 용매와 혼합된 이온성 액체에 황금을 혼합한 후 초음파 추출을 진행하고, 반용매를 첨가하여 타겟 플라보노이드의 과포화를 유도하였다. 상기 과포화로 유도된 침전물을 pH 2.53, pH 4.02, DW의 재결정용매에 용해시켜 재결정을 유도하였다. 바이칼린의 추출 수율 및 회수율을 측정하였고, 그 결과를 도 6에 나타내었다.After mixing gold with the ionic liquid mixed with each alcohol solvent, ultrasonic extraction was performed, and supersaturation of the target flavonoid was induced by adding an anti-solvent. Recrystallization was induced by dissolving the precipitate induced by supersaturation in a recrystallization solvent of pH 2.53, pH 4.02, DW. The extraction yield and recovery rate of baicalin were measured, and the results are shown in FIG. 6 .
도 6에 나타낸 바와 같이, 상기 3가지 재결정용매에 따른 바이칼린의 추출 수율 및 회수율을 측정한 결과, pH 2.53 용액에서만 재결정화가 유도된 것을 확인하였다. 또한, 추출용매로 70% 에탄올에 녹인 이온성 액체를 사용하였을 경우 61.10%로 최대 회수율을 나타내었다.As shown in Figure 6, as a result of measuring the extraction yield and recovery rate of baicalin according to the three recrystallization solvents, it was confirmed that recrystallization was induced only in the pH 2.53 solution. In addition, when the ionic liquid dissolved in 70% ethanol was used as the extraction solvent, the maximum recovery rate was 61.10%.
[실시예 7][Example 7]
생약으로부터 플라보노이드의 추출 및 분리Extraction and isolation of flavonoids from herbal medicines
7-1. 황금으로부터 플라보노이드의 추출7-1. Extraction of flavonoids from gold
황금으로부터 플라보노이드의 추출물을 얻기 위하여 건조시킨 황금 10g을 분쇄하고, 50mesh의 체로 걸러 300μm 입자 크기를 가진 생약을 골라내었다. 이온성 액체 1-에틸피리디늄 브로마이드를 3.5M의 농도로 70% 에탄올과 혼합하여 추출용매로 하였다. 300μm 입자의 생약 1g을 칭량하여 추출용매 10ml와 혼합하고, 효과적인 추출을 위해서 볼텍싱하여 섞어주었다. 이후 황금-이온성 액체 혼합물을 50°C 수조에 넣어 90분 동안 초음파 추출을 하였다. 90분 후에 600rpm에서 5분 동안 원심분리를 시키고, 상층의 추출액을 획득하였다.In order to obtain an extract of flavonoids from gold, 10 g of dried gold was ground, and crude drugs having a particle size of 300 μm were selected through a 50 mesh sieve. The ionic liquid 1-ethylpyridinium bromide was mixed with 70% ethanol at a concentration of 3.5M as an extraction solvent. 1 g of crude drug with 300 μm particles was weighed and mixed with 10 ml of extraction solvent, and mixed by vortexing for effective extraction. Then, the gold-ionic liquid mixture was placed in a 50 °C water bath and subjected to ultrasonic extraction for 90 minutes. After 90 minutes, centrifugation was performed at 600 rpm for 5 minutes, and an extract of the upper layer was obtained.
7-2. 플라보노이드의 1차 결정화7-2. Primary crystallization of flavonoids
상기 실시예 7-1에서 획득한 추출물에 8배 부피비의 아세토니트릴을 반용매로 넣고, 볼텍싱하여 섞어주었다. 추출물-아세토니트릴의 혼합물을 실온에서 1시간 동안 방치하여 과포화를 통한 용석 결정화를 유도한 후 600rpm에서 5분 동안 원심분리를 시키고, 상층액을 제거하여 1차 침전물인 고형분의 펠렛을 얻어내었다.An 8-fold volume ratio of acetonitrile was added as an anti-solvent to the extract obtained in Example 7-1, and mixed by vortexing. The extract-acetonitrile mixture was left at room temperature for 1 hour to induce soluble crystallization through supersaturation, and then centrifuged at 600 rpm for 5 minutes, and the supernatant was removed to obtain a solid pellet as the primary precipitate.
7-3. 플라보노이드의 재결정화7-3. Recrystallization of flavonoids
상기 실시예 7-2에서 획득한 1차 침전물을 0.1% 수용성 포름산을 재결정용매로 추가한 후 볼텍싱하여 섞어주었다. 1차 침전물-0.1% 수용성 포름산의 혼합물을 실온에서 24시간 동안 방치하여 재결정화를 유도하고, 최종 생성물인 2차 침전물이 가라앉도록 하였다. The primary precipitate obtained in Example 7-2 was mixed by vortexing after adding 0.1% water-soluble formic acid as a recrystallization solvent. A mixture of the primary precipitate and 0.1% aqueous formic acid was allowed to stand at room temperature for 24 hours to induce recrystallization, and the final product, the secondary precipitate, was allowed to settle.
7-4. 플라보노이드의 분리7-4. Isolation of flavonoids
최종 생성물을 얻어내기 위해서 상기 실시예 7-3에서 획득한 2차 침전물을 증류수로 씻어내고, 오븐에서 용매를 완전히 제거하여 건조시켜 98.00% 순도의 최종 생성물을 61.10%의 회수율로 추출액에서 얻어내었다.In order to obtain the final product, the secondary precipitate obtained in Example 7-3 was washed with distilled water, and the solvent was completely removed in an oven and dried to obtain a final product with a purity of 98.00% at a recovery rate of 61.10%.
[실시예 8] [Example 8]
황금 (The roots of The roots of gold Scutellaria baicalensisScutellaria baicalensis )으로부터 추출 및 분리된 플라보노이드의 구조 분석) Structural analysis of flavonoids extracted and isolated from
황금으로부터 분리된 플라보노이드를 1H-NMR을 통해 분석하였고, 그 결과를 도 7에 나타내었다.Flavonoids isolated from gold were analyzed through 1 H-NMR, and the results are shown in FIG. 7 .
도 7에 나타낸 바와 같이, 세척이 되지 않은 미정제 침전물은 이온성 액체를 포함하는 불순물의 피크를 나타내고, 증류수로 세척한 최종 침전물은 불순물의 피크를 나타내지 않았다. 따라서, 이온성 액체 등이 친수성 용매로 씻어내는 과정을 통해 제거되어 최종 플라보노이드가 수득될 수 있는 것을 확인하였다.As shown in FIG. 7 , the unwashed crude precipitate shows the peak of impurities including the ionic liquid, and the final precipitate washed with distilled water does not show the peak of the impurity. Therefore, it was confirmed that the final flavonoid could be obtained by removing the ionic liquid and the like through a process of washing with a hydrophilic solvent.
[실시예 9] [Example 9]
괴화 (The flowers of The flowers of Goehwa Sophora japonicaSophora japonica )로부터 추출 및 분리된 플라보노이드의 HPLC 분석) HPLC analysis of flavonoids extracted and isolated from
괴화로부터 분리된 플라보노이드를 HPLC를 통해 분석하였고, 그 결과를 도 8에 나타내었다.Flavonoids isolated from necrosis were analyzed by HPLC, and the results are shown in FIG. 8 .
도 8에 나타낸 바와 같이, 최종 이온성 액체를 포함한 불순물의 피크는 나타나지 않았고, UV에 의한 순도는 97.00% 이상으로 나타났다. 따라서, 이온성 액체 등이 친수성 용매로 씻어내는 과정을 통해 제거되어 최종 플라보노이드가 수득될 수 있는 것을 확인하였다.As shown in FIG. 8, the peak of impurities including the final ionic liquid did not appear, and the purity by UV was 97.00% or more. Therefore, it was confirmed that the final flavonoid could be obtained by removing the ionic liquid and the like through a process of washing with a hydrophilic solvent.
Claims (22)
(b) 상기 플라보노이드 추출용액에 반용매 (anti-solvent)를 첨가하여 1차 침전물을 수득하는 단계;
(c) 상기 1차 침전물에 재결정용매 (recrystallization solvent)를 첨가하여 2차 침전물을 수득하는 단계; 및
(d) 상기 2차 침전물을 여과하는 단계를 포함하는, 천연물로부터 플라보노이드를 추출 및 분리하는 방법.
(a) obtaining a flavonoid extract solution by adding an ionic liquid to a natural product;
(b) obtaining a primary precipitate by adding an anti-solvent to the flavonoid extract solution;
(c) obtaining a secondary precipitate by adding a recrystallization solvent to the primary precipitate; and
(d) a method for extracting and separating flavonoids from natural products, comprising filtering the secondary precipitate.
The method of claim 1, wherein the cation constituting the ionic liquid is morpholinium, imidazolium cation, oxazolium cation, piperidinium cation, pyrazinium cation, pyrazolium cation, pyridazininium cation, pyridinium Extraction and separation of flavonoids from natural products, characterized in that they are selected from the group consisting of cations, pyrimidinium cations, pyrrolidinium cations, pyrrolinium cations, pyrroleium cations, sulfonium cations, phosphonium cations and triazolium cations How to.
The method of claim 1, wherein the anion constituting the ionic liquid is ethyl sulfate anion, tosylate anion, methanesulfone anion, tetrafluoroborate anion, hexafluorophosphate anion, hexafluoroantimonate anion, tetrachloroaluminate, A method for extracting and separating flavonoids from natural products, characterized in that they are selected from the group consisting of bromide anions and chloride anions.
The method of claim 1, wherein the ionic liquid is 1-butyl-3-methylimidazolium bromide, 1-butyl-3-methylimidazolium chloride, 1-butyl-3-methylimidazolium tetrafluoroborate, 1-Butyl-3-methylimidazolium hexafluorophosphate, 1-methyl-3-ethylimidazolium bromide, 1-octyl-3-methylimidazolium bromide, 1-ethylpyridinium bromide, 1-ethyl- 3-methylimidazolium ethyl sulfate, 1-ethyl-3-methylimidazolium tetrafluoroborate, 1-ethyl-3-methylimidazolium tosylate, 1-ethyl-3-methylimidazolium methanesulfonate , 1-ethyl-3-methylimidazolium preplate, 1-ethyl-3-methylimidazolium dicyanamide, 1-butyl-3-methylimidazolium tetrafluoroborate, 1-butyl-3-methyl Midazolium bis(triplefluoromethylsulfonyl)imide, 1-butyl-3-methylimidazolium dicyanamide, 1-butyl-3-methylimidazolium triflate, 1-butyl-3-methylimida Zolium methanesulfonate, 1,3-didecyl-2-methylimidazolium bis(triplefluoromethylsulfonyl)imide, 1,3-diallylimidazolium tetrafluoroborate, 1-butyl-2, 3-dimethylimidazolium triflate, 1-butyl-2,3-dimethylimidazolium tetrafluoroborate, 1-(2-hydroxyethyl)-3-methylimidazolium tetrafluoroborate, 1-decyl -3-methylimidazolium tetrafluoroborate, 1-butyl-3-methylpyridinium tetrafluoroborate, 1-butyl-4-methylpyridinium tetrafluoroborate, 1-allyl-3-ethylimidazolium Tetrafluoroborate, 1-methyl-1propylpiperidinium bis(trifluoromethylsulfonyl)imide, methyltrioctylammonium bis(trifluoromethylsulfonyl)imide, butyltrimethylammonium bis(trifluoro A natural product, characterized in that it is selected from the group consisting of methylsulfonyl)imide, triethylsulfonium bis(trifluoromethylsulfonyl)imide and diethylmethylsulfonium bis(trifluoromethylsulfonyl)imide A method for extracting and isolating flavonoids from
The method of claim 4, wherein the ionic liquid is 1-butyl-3-methylimidazolium bromide, 1-butyl-3-methylimidazolium chloride, 1-butyl-3-methylimidazolium tetrafluoroborate, from the group consisting of 1-butyl-3-methylimidazolium hexafluorophosphate, 1-methyl-3-ethylimidazolium bromide, 1-octyl-3-methylimidazolium bromide, and 1-ethylpyridinium bromide A method for extracting and separating flavonoids from natural products, characterized in that selected.
The method of claim 1, wherein in step (a), the natural product and the ionic liquid are included in a weight ratio of 1:3 to 1:30.
The method for extracting and separating flavonoids from natural products according to claim 1, wherein the natural product and the ionic liquid are included in a weight ratio of 1:10 in step (a).
The method of claim 1, wherein in the step (a), an ionic liquid is added to the natural product to obtain a flavonoid extraction solution through ultrasonic extraction, and the ultrasonic extraction temperature is 0 to 60 ° C. How to extract and isolate.
The method of claim 1, wherein in the step (a), an ionic liquid is added to the natural product to obtain a flavonoid extraction solution through ultrasonic extraction, and the ultrasonic extraction temperature is 30 to 50 ° C. How to extract and isolate.
The method of claim 1, wherein in the step (a), an ionic liquid is added to the natural product to obtain a flavonoid extraction solution through ultrasonic extraction, and the ultrasonic extraction time is 120 minutes or less. how to separate.
The method of claim 1, wherein in step (a), an ionic liquid is added to the natural product to obtain a flavonoid extraction solution through ultrasonic extraction, and the ultrasonic extraction time is 90 minutes. How to.
The method for extracting and separating flavonoids from natural products according to claim 1, characterized in that an alcohol solvent is mixed with the ionic liquid.
13. The method for extracting and separating flavonoids from natural products according to claim 12, characterized in that the ionic liquid is mixed at a concentration of 6 M or less.
13. The method for extracting and separating flavonoids from natural products according to claim 12, wherein the ionic liquid is mixed at a concentration of 5M or less.
13. The method of claim 12, wherein the ionic liquid is mixed at a concentration of 1M to 4M.
The method for extracting and separating flavonoids from natural products according to claim 1, wherein the anti-solvent has a polarity less than methanol in the polarity table.
The flavonoid of claim 1, wherein the anti-solvent is selected from the group consisting of methyl acetate, methylene chloride, benzene, butanol, acetonitrile, acetone, ethanol, ethyl acetate, isopropyl alcohol, and chloroform. How to extract and isolate.
The method for extracting and separating flavonoids from natural products according to claim 1, wherein in step (b), the flavonoid extraction solution and the anti-solvent are included in a volume ratio of 1:3 to 1:30.
The method of claim 1, wherein the recrystallization solvent is selected from the group consisting of water, methanol, acetic acid, ethanol and formic acid.
The method of claim 1, wherein step (d) comprises: filtering the secondary precipitate under reduced pressure; washing the filtered precipitate with a hydrophilic solvent; and removing a solvent from the washed precipitate.
21. The method of claim 20, wherein the hydrophilic solvent is selected from water, methanol or ethanol.
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| US7595080B2 (en) | 2003-10-31 | 2009-09-29 | Rexgenebiotech Co., Ltd. | Method for preparing an extract of fruit of Sophora japonica containing isoflavone |
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| KR20060099690A (en) | 2005-03-14 | 2006-09-20 | 삼성전자주식회사 | Non-volatile semiconductor memory device and method of fabrication the same |
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