CN101891727A - Method for separating and extracting apigenin, acacetin-7-O-beta-D-glucoside and apigenin-7-O-beta-D-glucoside from dendranthema indicum - Google Patents
Method for separating and extracting apigenin, acacetin-7-O-beta-D-glucoside and apigenin-7-O-beta-D-glucoside from dendranthema indicum Download PDFInfo
- Publication number
- CN101891727A CN101891727A CN200910062199XA CN200910062199A CN101891727A CN 101891727 A CN101891727 A CN 101891727A CN 200910062199X A CN200910062199X A CN 200910062199XA CN 200910062199 A CN200910062199 A CN 200910062199A CN 101891727 A CN101891727 A CN 101891727A
- Authority
- CN
- China
- Prior art keywords
- apigenin
- glucoside
- beta
- dendranthema indicum
- extraction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 240000005250 Chrysanthemum indicum Species 0.000 title claims abstract description 33
- 235000018959 Chrysanthemum indicum Nutrition 0.000 title claims abstract description 31
- KZNIFHPLKGYRTM-UHFFFAOYSA-N apigenin Chemical compound C1=CC(O)=CC=C1C1=CC(=O)C2=C(O)C=C(O)C=C2O1 KZNIFHPLKGYRTM-UHFFFAOYSA-N 0.000 title claims abstract description 23
- XADJWCRESPGUTB-UHFFFAOYSA-N apigenin Natural products C1=CC(O)=CC=C1C1=CC(=O)C2=CC(O)=C(O)C=C2O1 XADJWCRESPGUTB-UHFFFAOYSA-N 0.000 title claims abstract description 23
- 235000008714 apigenin Nutrition 0.000 title claims abstract description 23
- 229940117893 apigenin Drugs 0.000 title claims abstract description 23
- KMOUJOKENFFTPU-QNDFHXLGSA-N apigenin 7-O-beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=C2C(=O)C=C(C=3C=CC(O)=CC=3)OC2=C1 KMOUJOKENFFTPU-QNDFHXLGSA-N 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title claims abstract description 21
- NLZCOTZRUWYPTP-UHFFFAOYSA-N acacetin-7-O-beta-D-galactoside Natural products C1=CC(OC)=CC=C1C(OC1=C2)=CC(=O)C1=C(O)C=C2OC1C(O)C(O)C(O)C(CO)O1 NLZCOTZRUWYPTP-UHFFFAOYSA-N 0.000 title abstract description 4
- 239000000284 extract Substances 0.000 claims abstract description 24
- 238000000926 separation method Methods 0.000 claims abstract description 17
- 238000000605 extraction Methods 0.000 claims abstract description 16
- 150000001875 compounds Chemical class 0.000 claims abstract description 13
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims abstract description 12
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000007788 liquid Substances 0.000 claims abstract description 12
- 239000003208 petroleum Substances 0.000 claims abstract description 6
- 238000002360 preparation method Methods 0.000 claims abstract description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 13
- 238000010898 silica gel chromatography Methods 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 239000000843 powder Substances 0.000 claims description 8
- 239000012141 concentrate Substances 0.000 claims description 7
- 238000003810 ethyl acetate extraction Methods 0.000 claims description 7
- 230000006837 decompression Effects 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 6
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 5
- 229940079593 drug Drugs 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 5
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 claims description 5
- 238000007654 immersion Methods 0.000 claims description 5
- 239000003921 oil Substances 0.000 claims description 5
- 238000001953 recrystallisation Methods 0.000 claims description 5
- 238000004587 chromatography analysis Methods 0.000 claims description 4
- 238000004458 analytical method Methods 0.000 claims description 2
- 238000010828 elution Methods 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- 239000000126 substance Substances 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 7
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 abstract description 5
- 238000004440 column chromatography Methods 0.000 abstract description 4
- 238000003912 environmental pollution Methods 0.000 abstract description 4
- 241000196324 Embryophyta Species 0.000 abstract description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 abstract 2
- 239000000377 silicon dioxide Substances 0.000 abstract 2
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 30
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Substances OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 16
- 238000006243 chemical reaction Methods 0.000 description 14
- 239000003153 chemical reaction reagent Substances 0.000 description 8
- 239000008103 glucose Substances 0.000 description 8
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 6
- 238000005160 1H NMR spectroscopy Methods 0.000 description 6
- -1 flavonoid compound Chemical class 0.000 description 5
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- 238000005903 acid hydrolysis reaction Methods 0.000 description 4
- 230000008034 disappearance Effects 0.000 description 4
- 229930182486 flavonoid glycoside Natural products 0.000 description 4
- CMOAHYOGLLEOGO-UHFFFAOYSA-N oxozirconium;dihydrochloride Chemical compound Cl.Cl.[Zr]=O CMOAHYOGLLEOGO-UHFFFAOYSA-N 0.000 description 4
- 241000723353 Chrysanthemum Species 0.000 description 3
- 235000007516 Chrysanthemum Nutrition 0.000 description 3
- 235000008495 Chrysanthemum leucanthemum Nutrition 0.000 description 3
- 244000192528 Chrysanthemum parthenium Species 0.000 description 3
- 235000000604 Chrysanthemum parthenium Nutrition 0.000 description 3
- 239000002024 ethyl acetate extract Substances 0.000 description 3
- 235000008384 feverfew Nutrition 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 238000004811 liquid chromatography Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 description 2
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 description 2
- 241000201295 Euphrasia Species 0.000 description 2
- 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 2
- FOGVNFMUZXDMTR-UHFFFAOYSA-N [Mg].Cl Chemical compound [Mg].Cl FOGVNFMUZXDMTR-UHFFFAOYSA-N 0.000 description 2
- 238000002425 crystallisation Methods 0.000 description 2
- 230000008025 crystallization Effects 0.000 description 2
- 238000002451 electron ionisation mass spectrometry Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 229930003944 flavone Natural products 0.000 description 2
- 150000002212 flavone derivatives Chemical class 0.000 description 2
- 235000011949 flavones Nutrition 0.000 description 2
- 229930003935 flavonoid Natural products 0.000 description 2
- 235000017173 flavonoids Nutrition 0.000 description 2
- 125000003147 glycosyl group Chemical group 0.000 description 2
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 description 2
- 238000004611 spectroscopical analysis Methods 0.000 description 2
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 2
- KMOUJOKENFFTPU-UHFFFAOYSA-N Apigenin-7-glucosid Natural products OC1C(O)C(O)C(CO)OC1OC1=CC(O)=C2C(=O)C=C(C=3C=CC(O)=CC=3)OC2=C1 KMOUJOKENFFTPU-UHFFFAOYSA-N 0.000 description 1
- 208000004429 Bacillary Dysentery Diseases 0.000 description 1
- 206010010774 Constipation Diseases 0.000 description 1
- ACBOFPQSBWBAQR-UHFFFAOYSA-N Cosmosiin Natural products OCC1OC(Oc2cc(O)c3C(=O)C=C(Oc3c2)c4cccc(O)c4)C(O)C(O)C1O ACBOFPQSBWBAQR-UHFFFAOYSA-N 0.000 description 1
- 208000004232 Enteritis Diseases 0.000 description 1
- 241000628997 Flos Species 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 206010017915 Gastroenteritis shigella Diseases 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 239000002260 anti-inflammatory agent Substances 0.000 description 1
- 230000001741 anti-phlogistic effect Effects 0.000 description 1
- KMOUJOKENFFTPU-JOZSIVFUSA-N apigenin-7-O-beta-D-glucopyranoside Natural products OC[C@@H]1O[C@@H](Oc2cc(O)c3c(c2)oc(cc3=O)-c2ccc(O)cc2)[C@H](O)[C@@H](O)[C@@H]1O KMOUJOKENFFTPU-JOZSIVFUSA-N 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 235000019504 cigarettes Nutrition 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 150000002215 flavonoids Chemical class 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- KEEWIHDTSNESJZ-UHFFFAOYSA-N poncirenin Natural products C1=CC(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(CO)O3)O)=CC(O)=C2C(=O)C1 KEEWIHDTSNESJZ-UHFFFAOYSA-N 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000011435 rock Substances 0.000 description 1
- 201000005113 shigellosis Diseases 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
Landscapes
- Saccharide Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a method for separating and extracting apigenin, acacetin-7-O-beta-D-glucoside and apigenin-7-O-beta-D-glucoside from dendranthema indicum, which belongs to a method for extracting chemical substances from a natural plant. The method is sequentially carried out according to the following steps of: 1. extract preparation; 2. extraction: sequentially extracting the extract of the dendranthema indicum by using petroleum ether, chloroform and ethyl acetate; and 3. separation and extraction: (1) carrying out normal-pressure and reduced-pressure silica column chromatography on the extracted part by using the ethyl acetate; and (2) separating fractions through a semi-preparative high-efficiency liquid chromatographic instrument to obtain the acacetin-7-O-beta-D-glucoside, the apigenin-7-O-beta-D-glucoside and the apigenin as compounds. In the method, the semi-preparative high-efficiency liquid chromatographic instrument is directly adopted for separation to obtain the three compounds after silica column chromatography is adopted, and the method has good separation effect, short time, high product purity and less environmental pollution.
Description
Technical field
The invention belongs to a kind of method of from natural phant, extracting chemical substance, be specifically related to separate the method that obtains apigenin, robinin-7-O-β-D-glucoside, apigenin-7-O-beta-D-glucoside from the natural phant Dendranthema indicum.
Background technology
Dendranthema indicum Dendranthema indicum (L.) Des Monl.var aromaticum Q.H.Liuet S.F.Zhang. is the new variant of composite family Chrysanthemum (Dendranthema), be the Shennongjia endemic plant, originate in Hubei Province's Shennongjiawooded Area, be distributed in the field on the open hillside that faces south of height above sea level more than 1200 meters, roadside, rock seam, shrubbery limit, complete stool has special strong fragrance, among the people with its flower, leaf dries in the shade and makes sachet.The west place in Hubei mountain area with Dendranthema indicum be used for anti-curing cold, diseases such as headache, bacillary dysentery, enteritis, constipation, coronary heart disease and hypertension, Dendranthema indicum also has bactericidal antiphlogistic, flat liver make eye bright, the loose effect of wind step-down.It can be used to extract essential oil or medicinal extract, is used for aspects such as medicine, beverage, cigarette and makeup.Studies show that Dendranthema indicum dispelling wind, heat-clearing, make eye bright, the main active ingredient of detoxicating functions is flavonoid compound.But it is at present less about the chemical substance fundamental research of Dendranthema indicum, in the existing relevant report, as yet not relevant for the report of finding apigenin, robinin-7-O-β-D-glucoside, apigenin-7-O-beta-D-glucoside to the Dendranthema indicum chemical ingredients.Existing separation about feverfew chemical ingredients apigenin, robinin-7-O-β-D-glucoside, apigenin-7-O-beta-D-glucoside, especially the separation method of Dendranthema indicum chemical ingredients adopts repeatedly after the silica gel wash-out more, adopt the stationary phase of costliness of other types and the column chromatography that consumes a large amount of organic reagents to separate again and obtain compound, this method environmental pollution is bigger.Carrying out in separation and purification and the research that structure is determined as Yang Yansong etc. to flavonoid activeconstituents in the Flos Matricariae chamomillae is that column chromatography is separated repeatedly, purifying obtains apigenin-7-O-beta-D-glucoside through polymeric amide, and its method separation cycle is long, consume that reagent is big, cost is high and big for environment pollution.Gu Yaohua etc. adopt repeatedly silica gel column chromatography to separate from the Bo chrysanthemum and obtain apigenin, apigenin-7-O-beta-D-glucoside, and its method separation cycle is long, products obtained therefrom purity is low, consume that reagent is big, cost is high and big for environment pollution.
Summary of the invention
Above-mentioned deficiency at prior art, the technical problem to be solved in the present invention provide a kind of from Dendranthema indicum the method for separation and Extraction apigenin, robinin-7-O-β-D-glucoside, apigenin-7-O-beta-D-glucoside, it have good separating effect, time short, obtain the product purity height, consume advantages such as organic reagent is few, environmental pollution is little.
Technical scheme of the present invention is: the method for separation and Extraction apigenin, robinin-7-O-β-D-glucoside, apigenin-7-O-beta-D-glucoside is carried out according to the following steps successively from Dendranthema indicum:
[1] preparation medicinal extract
(1), pulverized 20 mesh sieves with the Dendranthema indicum dried flower;
(2) Dendranthema indicum dried flower powder after 12 hours, is used 70% ethanol percolate extraction with 70% alcohol immersion, collect percolate;
(3) collect percolate 10 times, adopt heating in water bath or Rotary Evaporators that percolate is concentrated again to the crude drug amount, concentrate Dendranthema indicum medicinal extract;
[2] extraction
In Dendranthema indicum medicinal extract, add distilled water, make Dendranthema indicum medicinal extract in water, be suspended state, with sherwood oil, chloroform, ethyl acetate Dendranthema indicum medicinal extract is extracted successively, obtain 3 extract parts, keep the ethyl acetate extraction position;
[3] separation and Extraction
(1) above-mentioned ethyl acetate extraction position is carried out normal pressure or decompression 200-300 purpose silica gel column chromatography carries out chromatography, with petroleum ether-ethyl acetate, chloroform-methanol system repeatedly elution chromatography post, detect stream part with the thin layer analysis method, stream part is collected in the colour developing back;
(2) above-mentioned stream part is separated through half preparative high-performance liquid chromatographic instrument, obtain 3 kinds of stream parts after the separation, with 3 kinds of streams part concentrate respectively, drying and recrystallization, obtain compound robinin-7-O-β-D-glucoside, apigenin-7-O-beta-D-glucoside, apigenin respectively.
Method of the present invention adopts and directly adopts half preparative high-performance liquid chromatographic instrument to separate after the silica gel column chromatography to obtain above three compounds, and present method good separating effect, time be short, obtain the product purity height, consume that organic reagent is few, environmental pollution is little.
Embodiment
Below relevant technologies problem of the present invention is described in further detail.
Present method adopts silica gel column chromatography directly to adopt half preparative high-performance liquid chromatographic instrument to separate afterwards exactly with the improvements of similar document report maximum and obtains above three compounds.Its advantage is simple, the good separating effect of method, and the time is short, obtain the product purity height, significantly reduce the consumption of organic reagent, and environmental protection.Do not see about adopting normal pressure from feverfew, to separate the report that obtains apigenin, robinin-7-O-β-D-glucoside, apigenin-7-O-beta-D-glucoside in the prior art with reduce pressure silica gel column chromatography and half preparative high-performance liquid chromatographic method.Especially not about adopting normal pressure from the Dendranthema indicum plant, to separate the report that obtains apigenin, robinin-7-O-β-D-glucoside, apigenin-7-O-beta-D-glucoside with decompression silica gel column chromatography and half preparative high-performance liquid chromatographic method.
The applicant adopts the condition of half preparative high-performance liquid chromatographic to be: moving phase: methyl alcohol: water (63: 37), and flow velocity: 2.5mL/min, column temperature: 25 ℃, sample size: 0.5mL.The applicant has carried out relevant evaluation to 3 kinds of compounds that extracted, and the evaluation situation is made following brief description.
(1) robinin-7-O-β-D-glucoside
Faint yellow crystallization, 260～262 ℃ of mp.Hydrochloric acid-magnesium powder and Molish reaction are positive.Acid hydrolysis detects glucose.Prompt for flavonoid glycoside compound; In zirconates-Citric Acid color reaction, add zirconium oxychloride (ZrOCl
2) the apparent glassy yellow in back, add the yellow disappearance of Citric Acid again, so contain C in the molecule
5-OH exists, but does not have free C
3-OH exists.
1H-NMR[600MHz, DMSO-d
6]: 12.93 (1H, s, 5-OH), 8.08 (2H, d, J=7.8Hz, H-2 ' and 6 '), 7.15 (2H, d, J=7.8Hz, H-3 ' and 5 '), 6.91 (1H, s, H-3), 6.89 (1H, d, J=2.1Hz, H-8), 6.49 (1H, d, J=2.1Hz, H-6), 5.13～5.49 (4H, m, OH on the sugar), 5.10 (1H, d, J=7.2Hz, H-glucose anomeric proton is a beta configuration), 3.27-3.90 (5H, m, H on the sugar), 3.79 (3H, s, 4 '-OCH
3).
13C-NMR[600MHz，DMSO-d
6]：182.7(C-4)，163.7(C-7)，164.5(C-2)，161.8(C-4′)，157.6(C-9)，163.1(C-5)，128.9(C-2′and?6′)，123.0(C-1)，115.1(C-3′and?5′)，106.6(C-10)，100.1(C-6)，95.5(C-8)，55.9(C-OCH
3)。
According to above data and contrast bibliographical information, be accredited as robinin-7-O-β-D-glucoside (acacetin-7-O-β-D-glucopyranoside).Its chemical structure is as follows:
(2) apigenin-7-O-beta-D-glucoside
Yellow powder (MeOH), 226～228 ℃ of mp, hydrochloric acid-magnesium powder reaction and Molish reaction are all positive.Acid hydrolysis detects glucose, prompts for flavonoid glycoside compound; In zirconates-Citric Acid color reaction, add zirconium oxychloride (ZrOCl
2) the apparent glassy yellow in back, add the yellow disappearance of Citric Acid again, so contain C in the molecule
5-OH exists, but does not have free C
3-OH exists.
1H-NMR[600MHz, DMSO-d
6]: 7.78 (2H, d, J=9.0Hz, H-2 ' and 6 '), 6.83 (2H, d, J=9.0Hz, H-3 ' and 5 '), 6.71 (1H, s, H-3), 6.39 (1H, d, J=1.8, H-8), 6.39 (1H, d, J=1.8Hz, H-6,7 on A ring is replaced by glycosyl), 4.97 (1H, d, J=6.6Hz, H-glucose anomeric proton is a beta configuration), 3.39～3.84 (5H, m).
13C-NMR[600MHz，DMSO-d
6]：184.0(C-4)，164.8(C-7)，166.6(C-2)，162.8(C-4′)，158.9(C-9)，162.9(C-5)，129.6(C-2′，C-6′)，123.1(C-1′)，117.2(C-3′，C-5′)，107.1(C-10)，104.5(C-3)，101.2(C-6)，96.2(C-8)，78.6(C-5″)，77.9(C-3″)，74.8(C-2″)，71.3(C-4″)，62.5(C-6″)。
The apigenin-7-O-beta-D of above data and bibliographical information-glucoside basically identical is so authenticating compound is apigenin-7-O-beta-D-glucoside (apigenin-7-O-β-D-glucopyranoside).Its chemical structure is as follows:
(3) apigenin
Yellow powder (MeOH), 345～347 ℃ of mp are dissolved in methyl alcohol, ethanol, dilute alkaline soln.Hydrochloric acid-magnesium powder reaction is positive.The Molish reaction is negative, and prompting may be Flavone aglycone.
EI-MS?m/z：270，242，152，128，121，118。
1H-NMR(600MHz，DMSO-d
6)δ：7.71(2H，d，J＝7.8，H-2′and?H-6′)，6.83(2H，d，J＝7.8Hz，H-3′and?H-5′)，6.42(1H，s，H-3)，6.31(1H，d，J＝1.8Hz，H-8)，6.10(1H，d，J＝1.8Hz，H-6)。
13CNMR(600MHz，DMSO-d
6)δ：181.6(C-4)，165.9(C-2)，163.6(C-7)，161.2(C-4′)，161.4(C-9)，157.6(C-5)，128.5(C-2′and?6′)，121.3(C-1′)，116.1(C-3′and?5′)，103.2(C-3)，102.8(C-10)，99.3(C-6)，94.3(C-8)。
Above spectroscopic data and apigenin pertinent literature report data basically identical are so the deduction compound is apigenin (apigenin).Its chemical structure is as follows:
The present invention adopt and directly adopt good separating effect, time weak point, products obtained therefrom purity height after the silica gel column chromatography, consume half few preparative high-performance liquid chromatographic of organic reagent first from Dendranthema indicum separation obtain apigenin, robinin-7-O-β-D-glucoside, apigenin-7-O-beta-D-glucoside.Present method adopts silica gel column chromatography directly to adopt half preparative high-performance liquid chromatographic instrument to separate afterwards exactly with the improvements of similar document report maximum and obtains above three compounds.And adopting normal pressure to separate the report that obtains apigenin, robinin-7-O-β-D-glucoside, apigenin-7-O-beta-D-glucoside with reduce pressure silica gel column chromatography and half preparative high-performance liquid chromatographic method from feverfew does not see.Simple, the good separating effect of extraction process, the time is short, obtain product purity height, organic reagent consumption less, low cost, and environmental protection is for the chemical substance basis of research Dendranthema indicum provides certain reference frame.
Embodiment 1
The Dendranthema indicum dried flower, pulverized 20 mesh sieves, get 3 kilograms, with the diacolation extraction after 12 hours of 70% alcohol immersion, collect percolate 10 times to the crude drug amount, reclaim solvent and get medicinal extract, with an amount of distilled water medicinal extract is suspended, use sherwood oil successively, chloroform, ethyl acetate extraction obtains three extract parts, then ethyl acetate extract is carried out normal pressure and decompression 200-300 purpose silica gel column chromatography, with petroleum ether-ethyl acetate, chloroform-methanol system repeatedly wash-out for detecting means, is collected stream part with TLC, separate through half preparative high-performance liquid chromatographic instrument, concentrate, drying and recrystallization.Chromatographic condition: moving phase: methyl alcohol: water (63: 37), flow velocity: 2.5mL/min, column temperature: 25 ℃, sample size: 0.5mL.
Identify: faint yellow crystallization, 260～262 ℃ of mp.Hydrochloric acid-magnesium powder and Molish reaction are positive.Acid hydrolysis detects glucose.Prompt for flavonoid glycoside compound; In zirconates-Citric Acid color reaction, add zirconium oxychloride (ZrOCl
2) the apparent glassy yellow in back, add the yellow disappearance of Citric Acid again, so contain C in the molecule
5-OH exists, but does not have free C
3-OH exists.
1H-NMR[600MHz, DMSO-d
6]: 12.93 (1H, s, 5-OH), 8.08 (2H, d, J=7.8Hz, H-2 ' and 6 '), 7.15 (2H, d, J=7.8Hz, H-3 ' and 5 '), 6.91 (1H, s, H-3), 6.89 (1H, d, J=2.1Hz, H-8), 6.49 (1H, d, J=2.1Hz, H-6), 5.13～5.49 (4H, m, OH on the sugar), 5.10 (1H, d, J=7.2Hz, H-glucose anomeric proton is a beta configuration), 3.27-3.90 (5H, m, H on the sugar), 3.79 (3H, s, 4 '-OCH
3).
13C-NMR[600MHz，DMSO-d
6]：182.7(C-4)，163.7(C-7)，164.5(C-2)，161.8(C-4′)，157.6(C-9)，163.1(C-5)，128.9(C-2′and?6′)，123.0(C-1)，115.1(C-3′and?5′)，106.6(C-10)，100.1(C-6)，95.5(C-8)，55.9(C-OCH
3)。
According to above data and contrast bibliographical information, be accredited as robinin-7-O-β-D-glucoside.
Embodiment 2
The Dendranthema indicum dried flower, pulverized 20 mesh sieves, get 3 kilograms, with the diacolation extraction after 12 hours of 70% alcohol immersion, collect percolate 10 times to the crude drug amount, reclaim solvent and get medicinal extract, with an amount of distilled water medicinal extract is suspended, use sherwood oil, chloroform, ethyl acetate extraction successively, obtain three extract parts, then ethyl acetate extract is carried out normal pressure and decompression silica gel column chromatography, with petroleum ether-ethyl acetate, chloroform-methanol system repeatedly wash-out, for detecting means, collect stream part with TLC, separate through half preparative high-performance liquid chromatographic instrument, concentrate, drying and recrystallization.Chromatographic condition: moving phase: methyl alcohol: water (63: 37), flow velocity: 2.5mL/min, column temperature: 25 ℃, sample size: 0.5mL.
Identify: yellow powder (MeOH), 226～228 ℃ of mp, hydrochloric acid-magnesium powder reaction and Molish reaction are all positive.Acid hydrolysis detects glucose, prompts for flavonoid glycoside compound; In zirconates-Citric Acid color reaction, add zirconium oxychloride (ZrOCl
2) the apparent glassy yellow in back, add the yellow disappearance of Citric Acid again, so contain C in the molecule
5-OH exists, but does not have free C
3-OH exists.
1H-NMR[600MHz, DMSO-d
6]: 7.78 (2H, d, J=9.0Hz, H-2 ' and 6 '), 6.83 (2H, d, J=9.0Hz, H-3 ' and 5 '), 6.71 (1H, s, H-3), 6.39 (1H, d, J=1.8, H-8), 6.39 (1H, d, J=1.8Hz, H-6,7 on A ring is replaced by glycosyl), 4.97 (1H, d, J=6.6Hz, H-glucose anomeric proton is a beta configuration), 3.39～3.84 (5H, m).
13C-NMR[600MHz，DMSO-d
6]：184.0(C-4)，164.8(C-7)，166.6(C-2)，162.8(C-4′)，158.9(C-9)，162.9(C-5)，129.6(C-2′，C-6′)，123.1(C-1′)，117.2(C-3′，C-5′)，107.1(C-10)，104.5(C-3)，101.2(C-6)，96.2(C-8)，78.6(C-5″)，77.9(C-3″)，74.8(C-2″)，71.3(C-4″)，62.5(C-6″)。
The apigenin-7-O-beta-D of above data and bibliographical information-glucoside basically identical, authenticating compound are apigenin-7-O-beta-D-glucoside.
Embodiment 3
The Dendranthema indicum dried flower, pulverized 20 mesh sieves, get 3 kilograms, with the diacolation extraction after 12 hours of 70% alcohol immersion, collect percolate 10 times to the crude drug amount, reclaim solvent and get medicinal extract, medicinal extract is suspended, use sherwood oil successively with an amount of distilled water, chloroform, ethyl acetate extraction, obtain three extract parts, then ethyl acetate extract is carried out normal pressure and decompression 200-300 purpose silica gel column chromatography, with petroleum ether-ethyl acetate, chloroform-methanol system repeatedly wash-out, with TLC for detecting means, collect stream part, separate through half preparative high-performance liquid chromatographic, collection stream part, a stream part warp concentrates, drying and recrystallization.Chromatographic condition: moving phase: methyl alcohol: water (63: 37), flow velocity: 2.5mL/min, column temperature: 25 ℃, sample size: 0.5mL.
Identify: yellow powder (MeOH), 345～347 ℃ of mp are dissolved in methyl alcohol, ethanol, dilute alkaline soln.Hydrochloric acid-magnesium powder reaction is positive.The Molish reaction is negative, and prompting may be Flavone aglycone.
EI-MS?m/z：270，242，152，128，121，118。
1H-NMR(600MHz，DMSO-d
6)δ：7.71(2H，d，J＝7.8，H-2′and?H-6′)，6.83(2H，d，J＝7.8Hz，H-3′and?H-5′)，6.42(1H，s，H-3)，6.31(1H，d，J＝1.8Hz，H-8)，6.10(1H，d，J＝1.8Hz，H-6)。
13CNMR(600MHz，DMSO-d
6)δ：181.6(C-4)，165.9(C-2)，163.6(C-7)，161.2(C-4′)，161.4(C-9)，157.6(C-5)，128.5(C-2′and?6′)，121.3(C-1′)，116.1(C-3′and?5′)，103.2(C-3)，102.8(C-10)，99.3(C-6)，94.3(C-8)。
Above spectroscopic data and apigenin pertinent literature report data basically identical, authenticating compound is an apigenin.
Claims (1)
1. the method for a separation and Extraction apigenin from Dendranthema indicum, robinin-7-O-β-D-glucoside, apigenin-7-O-beta-D-glucoside, it is characterized in that: described separating and extracting method carries out according to the following steps successively:
[1] preparation medicinal extract
(1), pulverized 20 mesh sieves with the Dendranthema indicum dried flower;
(2) Dendranthema indicum dried flower powder after 12 hours, is used 70% ethanol percolate extraction with 70% alcohol immersion, collect percolate;
(3) collect percolate 10 times, adopt heating in water bath or Rotary Evaporators that percolate is concentrated again to the crude drug amount, concentrate Dendranthema indicum medicinal extract;
[2] extraction
In Dendranthema indicum medicinal extract, add distilled water, make Dendranthema indicum medicinal extract in water, be suspended state, with sherwood oil, chloroform, ethyl acetate Dendranthema indicum medicinal extract is extracted successively, obtain 3 extract parts, keep the ethyl acetate extraction position;
[3] separation and Extraction
(1) above-mentioned ethyl acetate extraction position is carried out normal pressure or decompression 200-300 purpose silica gel column chromatography carries out chromatography, with petroleum ether-ethyl acetate, chloroform-methanol system repeatedly elution chromatography post, detect stream part with the thin layer analysis method, stream part is collected in the colour developing back;
(2) above-mentioned stream part is separated through half preparative high-performance liquid chromatographic instrument, obtain 3 kinds of stream parts after the separation, with 3 kinds of streams part concentrate respectively, drying and recrystallization, obtain compound robinin-7-O-β-D-glucoside, apigenin-7-O-beta-D-glucoside, apigenin respectively.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910062199XA CN101891727A (en) | 2009-05-22 | 2009-05-22 | Method for separating and extracting apigenin, acacetin-7-O-beta-D-glucoside and apigenin-7-O-beta-D-glucoside from dendranthema indicum |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910062199XA CN101891727A (en) | 2009-05-22 | 2009-05-22 | Method for separating and extracting apigenin, acacetin-7-O-beta-D-glucoside and apigenin-7-O-beta-D-glucoside from dendranthema indicum |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101891727A true CN101891727A (en) | 2010-11-24 |
Family
ID=43101102
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200910062199XA Pending CN101891727A (en) | 2009-05-22 | 2009-05-22 | Method for separating and extracting apigenin, acacetin-7-O-beta-D-glucoside and apigenin-7-O-beta-D-glucoside from dendranthema indicum |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101891727A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102477054A (en) * | 2010-11-25 | 2012-05-30 | 苏州宝泽堂医药科技有限公司 | Robinin preparation method |
| CN102703352A (en) * | 2012-06-03 | 2012-10-03 | 南京工业大学 | Bacterial strain and method for preparing glucosyl group apigenin by glycosylation in nonaqueous phase |
| CN106822165A (en) * | 2015-11-24 | 2017-06-13 | 石河子大学医学院第附属医院 | The purposes of the O glucuronides of robinin 7 |
| CN107586311A (en) * | 2017-07-25 | 2018-01-16 | 贵州维康子帆药业股份有限公司 | The method that the C β D glucosides of robinin 6 are extracted in creeping oxalis |
| CN108524574A (en) * | 2018-05-22 | 2018-09-14 | 武汉轻工大学 | A kind of Dendranthema indicum extract and application |
| CN115260144A (en) * | 2022-09-28 | 2022-11-01 | 山东省中医药研究院 | Method for quickly preparing acacetin based on roasting and hydrolyzing herba cephalanoploris |
-
2009
- 2009-05-22 CN CN200910062199XA patent/CN101891727A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102477054A (en) * | 2010-11-25 | 2012-05-30 | 苏州宝泽堂医药科技有限公司 | Robinin preparation method |
| CN102703352A (en) * | 2012-06-03 | 2012-10-03 | 南京工业大学 | Bacterial strain and method for preparing glucosyl group apigenin by glycosylation in nonaqueous phase |
| CN102703352B (en) * | 2012-06-03 | 2013-05-15 | 南京工业大学 | Bacterial strain and method for preparing glucosyl group apigenin by glycosylation in nonaqueous phase |
| CN106822165A (en) * | 2015-11-24 | 2017-06-13 | 石河子大学医学院第附属医院 | The purposes of the O glucuronides of robinin 7 |
| CN107586311A (en) * | 2017-07-25 | 2018-01-16 | 贵州维康子帆药业股份有限公司 | The method that the C β D glucosides of robinin 6 are extracted in creeping oxalis |
| CN107586311B (en) * | 2017-07-25 | 2018-10-09 | 贵州维康子帆药业股份有限公司 | The method that robinin -6-C- β-D-Glucose glycosides is extracted in creeping oxalis |
| CN108524574A (en) * | 2018-05-22 | 2018-09-14 | 武汉轻工大学 | A kind of Dendranthema indicum extract and application |
| CN115260144A (en) * | 2022-09-28 | 2022-11-01 | 山东省中医药研究院 | Method for quickly preparing acacetin based on roasting and hydrolyzing herba cephalanoploris |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101353363B (en) | Method for separating and purifying Momordica grosvenori leaf chromocor compound by high-speed countercurrent chromatography and products thereof | |
| CN104257715B (en) | Herba Artemisiae extract and its preparation method and application | |
| CN101891727A (en) | Method for separating and extracting apigenin, acacetin-7-O-beta-D-glucoside and apigenin-7-O-beta-D-glucoside from dendranthema indicum | |
| CN101899070A (en) | The preparation method of flavonoid glycoside in the compression leg sharp separation cake of camellia oleifera seeds in a kind of | |
| CN109879844A (en) | The extraction separation method of seven kinds of flavonoids in Sabia parviflora Wall.ex Roxb | |
| CN107759648A (en) | A kind of method that Hyperoside and isoquercitrin are isolated and purified from Golden flower | |
| CN110346462A (en) | Leaf of Moringa UPLC fingerprint and its UPLC finger-print | |
| CN102372754A (en) | Method for preparing specnuezhenide | |
| CN101648937B (en) | Isoflavones compound and preparation and application thereof | |
| CN104761596A (en) | Method for preparing anthocyanin standard substance | |
| CN101941961B (en) | Method for extracting and separating kaempferol from impatiens balsamina | |
| CN109942649A (en) | A kind of indoles glycosides compound and its purification methods and uses | |
| CN102260307A (en) | Method for preparing specnuezhenide | |
| CN104262316B (en) | A kind of flavonoid compound and its preparation method and application | |
| CN111620917B (en) | Isovitexin-2' -O-beta-D-glucopyranoside, and preparation method and application thereof | |
| CN101012257A (en) | Extraction and separation method for rhubarb willow leaf flavone component | |
| CN101407536A (en) | Process for preparing high-purity asiaticoside by solvent crystallization | |
| CN101648959B (en) | Coumaronochromones compound and preparation and application thereof | |
| CN1305870C (en) | Novel flavane derivative and its preparation method and uses | |
| CN102399207B (en) | Extraction method of ginkgetin or isoginkgetin | |
| CN107722087B (en) | Gynostemma pentaphylla flavonoid compound, preparation method thereof and application thereof in antitumor drugs | |
| CN101830997B (en) | Method for preparing reference substance of triterpenoid saponin in northeast clematis | |
| CN104592185A (en) | Method for extracting quercetin from eleocharis tuberosa peels | |
| CN105713005B (en) | A kind of extraction separation method of corymbose hedyotis herb middle ear humulone A | |
| CN104650164A (en) | Method for preparing active flavonoid glycoside monomers from pepper leaf |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20101124 |