CN101265247A - Method for separating effective constituent butenolide II from astraolylis lancea formalyrata volatile oil - Google Patents
Method for separating effective constituent butenolide II from astraolylis lancea formalyrata volatile oil Download PDFInfo
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- CN101265247A CN101265247A CNA2008100607964A CN200810060796A CN101265247A CN 101265247 A CN101265247 A CN 101265247A CN A2008100607964 A CNA2008100607964 A CN A2008100607964A CN 200810060796 A CN200810060796 A CN 200810060796A CN 101265247 A CN101265247 A CN 101265247A
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- volatile oil
- atractylenolide
- ethyl acetate
- rhizoma atractylodis
- atractylodis macrocephalae
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- 238000000034 method Methods 0.000 title claims abstract description 27
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- 239000000470 constituent Substances 0.000 title description 2
- SQEBMLCQNJOCBG-HVHJFMEUSA-N (5s)-3-(hydroxymethyl)-5-methoxy-4-methyl-5-[(e)-2-phenylethenyl]furan-2-one Chemical compound C=1C=CC=CC=1/C=C/[C@]1(OC)OC(=O)C(CO)=C1C SQEBMLCQNJOCBG-HVHJFMEUSA-N 0.000 title 1
- 241001131629 Lancea Species 0.000 title 1
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- 239000013078 crystal Substances 0.000 claims abstract description 12
- 238000004440 column chromatography Methods 0.000 claims abstract description 10
- ZTVSGQPHMUYCRS-SWLSCSKDSA-N (4as,8as)-3,8a-dimethyl-5-methylidene-4a,6,7,8-tetrahydro-4h-benzo[f][1]benzofuran-2-one Chemical compound C=C([C@@H]1C2)CCC[C@]1(C)C=C1C2=C(C)C(=O)O1 ZTVSGQPHMUYCRS-SWLSCSKDSA-N 0.000 claims description 26
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- 238000010828 elution Methods 0.000 claims description 3
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- OQYBLUDOOFOBPO-KCQAQPDRSA-N Atractylenolide II Chemical compound C=C([C@@H]1C2)CCC[C@]1(C)C[C@H]1C2=C(C)C(=O)O1 OQYBLUDOOFOBPO-KCQAQPDRSA-N 0.000 abstract description 4
- OQYBLUDOOFOBPO-YDHLFZDLSA-N atractylenolide II Natural products CC1=C2C[C@H]3C(=C)CCC[C@@]3(C)C[C@@H]2OC1=O OQYBLUDOOFOBPO-YDHLFZDLSA-N 0.000 abstract description 4
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 abstract description 3
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- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
Abstract
The invention discloses a method for separating effective component, atractylenolide II, from Rhizoma Atractylodis Macrocephalae volatile oil. The method includes pulverizing rhizome of dried Rhizoma Atractylodis Macrocephalae, extracting in medium and low polar solvents, and concentrating to obtain tan volatile oil; standing for a certain time, extracting the volatile oil with mixed solution prepared from n-hexane, ethyl acetate, ethanol and water at a certain ratio, collecting upper layer phase, and concentrating to obtain the refined volatile oil; and gradient eluting with mixed solvent of petroleum ether and ethyl acetate at a certain ratio with column chromatography, collecting correspondent fraction, concentrating, and recycling solvent to obtain atractylenolide II. The method performs pretreatment on Rhizoma Atractylodis Macrocephalae volatile oil coarse product by extracting and oxidizing before separating through column chromatography to have simple process and low cost, increased yield and purity of atractylenolide II, and avoids the disadvantages in the conventional post-treatment steps such as crystal loss caused by recrystallization, etc.
Description
Technical field
The invention belongs to the Chinese medicine pharmaceutical field, relate in particular to a kind of from Rhizoma Atractylodis Macrocephalae volatile oil the method for separating effective ingredient atractylenolide I.
Background technology
The bighead atractylodes rhizome (Atractylodes macrocephala koidz) is called in art, winter art, Zhejiang art etc., is per nnial herb, is composite family Atractylodes medicinal plant, and its rhizome can be used as medicine, and the beginning is stated from Shennong's Herbal, classifies as top grade.The bighead atractylodes rhizome is warm in nature, and it is sweet, bitter to distinguish the flavor of, and returns spleen, stomach warp, and invigorating the spleen and benefiting QI, eliminating dampness Li Shui, hidroschesis, antiabortive function are arranged.Main product is in Zhejiang, Anhui and other places, with Yinxian County, Zhejiang, area, Xinchang output maximum.Be mainly used in insufficiency of the spleen food less, maldigestion, abdominal distension have loose bowels, lassitude, phlegm and retained fluid are dizzyly throbbed with fear, oedema, spontaneous perspiration, threatened abortion.Modern pharmacological research shows, effect such as the bighead atractylodes rhizome protects the liver, cholagogic, diuresis, antibacterial, anti-inflammatory, antitumor, strengthening immunity, step-down, hypoglycemic, anticoagulation, strong, calmness.
The chemical ingredients of the bighead atractylodes rhizome is mainly Rhizoma Atractylodis Macrocephalae volatile oil, and in the volatile oil content maximum be atractylone, atractylodes lactone class material.Wherein, atractylone has hypotensive activity, atractylenolide, II are effective content of anti inflammation, atractylenolide II) has antitumous effect (Endo K, Toguchi T, Toguchi F, et al.Antiinflammatoryprinciples of Atractylodes.Chem.Pharm.Bull., 1979,27:2954-2958.).This constituents also has to be regulated gastrointestinal function and promotes the function that nutritive substance attracts.
Atractylenolide I (atractylenoide II) belongs to sesquiterpene lactones class chemicals, and molecular formula is C
15H
18O
2, molecular structural formula is:
Atractylenolide I is colourless needle, is dissolved in organic solvents such as methyl alcohol, ethanol, ethyl acetate, is insoluble to low polar solvents such as sherwood oil, normal hexane.
Atractylenolide I in the bighead atractylodes rhizome content only 0.07% (Li Wei. the active substance of the bighead atractylodes rhizome and quality approach. Nanjing: Nanjing University of Traditional Chinese Medicine, 2001,38.), and separate relatively difficulty.The normal separation method that adopts of prior art is that high speed adverse current chromatogram separates (Chunxia Zhao, Chaohong He.Preparative isolation and purificationof atractyl on and atractylenolide III from the Chinese medicinal plant Atractylodesmacrocephala by high-speed counter-current chromatography.J.Sep.Sci., 2006,29:1630-1636.) separate with column chromatography chromatogram (Li Wei. the active substance of the bighead atractylodes rhizome and quality approach. Nanjing: Nanjing University of Traditional Chinese Medicine, 2001,29-79.), but the former present stage suitability for industrialized production acquire a certain degree of difficulty, the latter generally need just can obtain purer material after follow-up means such as recrystallization are separated, this is possible loss part material again, moreover, separate the material yield with column chromatography, purity is all lower.
Summary of the invention
The objective of the invention is at the deficiencies in the prior art, provide a kind of from Rhizoma Atractylodis Macrocephalae volatile oil the method for separating effective ingredient atractylenolide I.
The objective of the invention is to be achieved through the following technical solutions: a kind of from Rhizoma Atractylodis Macrocephalae volatile oil the method for separating effective ingredient atractylenolide I, exsiccant bighead atractylodes rhizome rhizome is pulverized, powder with in, the solvent leaching of low isopolarity, concentrate, tawny volatile oil crude extract; The volatile oil crude extract of gained is placed for some time, treat its exhaustive oxidation after, with the mixed extractant solvent of normal hexane-ethyl acetate, alcohol and water, get phase by 4: 0～1: 1～4: 1 arbitrary proportion preparation, concentrate, get purified volatile oil; Purified volatile oil separates with the column chromatography chromatogram method, again with petroleum ether-ethyl acetate by 40～150: the mixed solvent of 1 arbitrary proportion preparation carries out gradient elution, collects corresponding flow point; Concentrate, reclaim solvent, get final product colourless acicular crystal atractylenolide I.
Further, described powder concentrates with air distillation or distillation under vacuum with the mixed solvent leaching of petroleum ether-ethyl acetate by 0～40: 1 arbitrary proportion preparation, gets tawny volatile oil crude extract; Described leaching method is that ultrasonic method, cold soaking, heat are carried, diacolation or Soxhlet are extracted.
Further, got mutually after, again with down mutually washing go up phase, concentrate with air distillation or distillation under vacuum, purified volatile oil.
The invention has the beneficial effects as follows: this method before column chromatography earlier to the Rhizoma Atractylodis Macrocephalae volatile oil crude product extract, pre-treatment step such as oxidation, simple to operate, cost is low, and removed the bigger composition of Rhizoma Atractylodis Macrocephalae volatile oil Semi-polarity, improved the wherein content of atractylenolide I, thereby can improve its yield and purity.
Description of drawings
Fig. 1 is the operational flowchart of the present invention's method of separating effective ingredient atractylenolide I from Rhizoma Atractylodis Macrocephalae volatile oil;
Fig. 2 is the column chromatography chromatogram figure of the present invention's method of separating effective ingredient atractylenolide I from Rhizoma Atractylodis Macrocephalae volatile oil.
Embodiment
Describe the present invention in detail with specific embodiment with reference to the accompanying drawings below, it is more obvious that purpose of the present invention and effect will become.
As shown in Figure 1, the method for the present invention separating effective ingredient atractylenolide I from Rhizoma Atractylodis Macrocephalae volatile oil may further comprise the steps:
One, exsiccant bighead atractylodes rhizome rhizome is pulverized, powder with in, the solvent leaching of low isopolarity, concentrate, tawny volatile oil.
Commercially available bighead atractylodes rhizome rhizome is placed on shady and cool place nature dries the back pulverizing, powder with petroleum ether-ethyl acetate in 0～40: the leaching of mixed solvent that 1 ratio is made into, leaching method comprises that ultrasonic method, cold soaking, heat are carried, diacolation or Soxhlet are extracted, concentrate with air distillation or distillation under vacuum, get tawny volatile oil.
Two, the Rhizoma Atractylodis Macrocephalae volatile oil crude extract placement for some time that step 1 is obtained with the mixed extractant solvent volatile oil that normal hexane-ethyl acetate, alcohol and water is prepared by a certain percentage, is got phase after treating its exhaustive oxidation, concentrates, and gets purified volatile oil.
The Rhizoma Atractylodis Macrocephalae volatile oil crude extract that step 1 is obtained is placed after for some time or heating make its exhaustive oxidation, pressed 4: 1: 4 with normal hexane-ethyl acetate, alcohol and water: 1 to 4: 0: 1: the mixed extractant solvent volatile oil of 1 arbitrary proportion preparation, get phase, go up the phase several times with clean following washing mutually again, concentrate with air distillation or distillation under vacuum, get purified volatile oil.
Three, the volatile oil after refining separates with the column chromatography chromatogram method, carries out gradient elution with the petroleum ether-ethyl acetate mixed solvent of certain proportioning, collects corresponding flow point.
Select the high chromatography column more than 10 times for the post footpath of post during column chromatography for use, the particle diameter of used silica gel is a 60-100 order or littler, and whether the silica gel heat-activated all can; Be 200: 1 or littler mixed solvent (gradient was less than 200: the 1) removal of impurities of polarity in gradient at first during chromatography, use the mixed solvent of the arbitrary proportion preparation of petroleum ether-ethyl acetate mixed solvent between 150: 1 to 40: 1 to carry out wash-out again with the petroleum ether-ethyl acetate mixed solvent; Eluent speed can be chosen wantonly below 10ml/min during removal of impurities, and speed should be 5ml/min or littler during wash-out, and collects corresponding flow point.
Four, concentrate, reclaim solvent, get final product colourless acicular crystal atractylenolide I.
Concentrate with air distillation, distillation under vacuum, reclaim solvent, or solvent is volatilized naturally, get final product colourless acicular crystal atractylenolide I.
Embodiment 1:
Bighead atractylodes rhizome rhizome is dried naturally, is ground into bighead atractylodes rhizome meal, with the ethyl acetate of 5 times of volumes leaching 2 times (60 ℃ water bath with thermostatic control, mixing speed 500r/min, each 1h), concentrate tawny volatile oil.Placement for some time uses normal hexane-ethyl acetate, alcohol and water in 4: 1: 4 after making its complete oxidation: the mixing solutions extraction of 1 ratio preparation, get phase, and concentrate, get refined volatile oil.Volatile oil after refining separates in the chromatography column of Φ 1.0 * 40cm with 60-100 purpose silica gel, at first adopting the petroleum ether-ethyl acetate mixed solvent is that 300: 1, flow velocity are removed impurity peaks during for 5ml/min in gradient, be that 50: 1, flow velocity are carried out wash-out during for 1ml/min with the petroleum ether-ethyl acetate mixed solvent in gradient again, collect corresponding flow point, concentrate, reclaim solvent, get final product colourless acicular crystal atractylenolide I, about 1.6% (is benchmark with the volatile oil applied sample amount) of yield, purity about 95.1%.
Embodiment 2:
Bighead atractylodes rhizome rhizome is dried naturally, be ground into bighead atractylodes rhizome meal, 2 (60 ℃ the waters bath with thermostatic control of mixed solvent leaching that are made in 1: 1 ratio with the petroleum ether-ethyl acetate of 5 times of volumes, mixing speed 500r/min, each 1h), place after for some time makes its complete oxidation, with normal hexane-ethyl acetate, alcohol and water in 4: 1: 4: the mixing solutions extraction of 1 ratio preparation, get phase, concentrate, get refined volatile oil.Volatile oil after refining separates in the chromatography column of Φ 1.0 * 40cm with 60-100 purpose silica gel, at first adopting the petroleum ether-ethyl acetate mixed solvent is that 300: 1, flow velocity are removed impurity peaks during for 5ml/min in gradient, be that 50: 1, flow velocity are carried out wash-out during for 1ml/min with the petroleum ether-ethyl acetate mixed solvent in gradient again, collect corresponding flow point, concentrate, reclaim solvent, get final product colourless acicular crystal atractylenolide I, about 1.9% (is benchmark with the volatile oil applied sample amount) of yield, purity about 95.2%.
Embodiment 3:
Bighead atractylodes rhizome rhizome is dried naturally, be ground into bighead atractylodes rhizome meal, 2 (60 ℃ the waters bath with thermostatic control of mixed solvent leaching that are made in 40: 1 ratio with the petroleum ether-ethyl acetate of 5 times of volumes, mixing speed 500r/min, each 1h), place after for some time makes its complete oxidation, with normal hexane-ethyl acetate, alcohol and water in 4: 1: 4: the mixing solutions extraction of 1 ratio preparation, get phase, concentrate, get refined volatile oil.Volatile oil after refining separates in the chromatography column of Φ 1.0 * 40cm with 60-100 purpose silica gel, at first adopting the petroleum ether-ethyl acetate mixed solvent is that 300: 1, flow velocity are removed impurity peaks during for 5ml/min in gradient, be that 50: 1, flow velocity are carried out wash-out during for 1ml/min with the petroleum ether-ethyl acetate mixed solvent in gradient again, collect corresponding flow point, concentrate, reclaim solvent, get final product colourless acicular crystal atractylenolide I, about 1.9% (is benchmark with the volatile oil applied sample amount) of yield, purity about 95.4%.
Embodiment 4:
Bighead atractylodes rhizome rhizome is dried naturally, be ground into bighead atractylodes rhizome meal, 2 (60 ℃ the waters bath with thermostatic control of mixed solvent leaching that are made in 40: 1 ratio with the petroleum ether-ethyl acetate of 5 times of volumes, mixing speed 500r/min, each 1h), place after for some time makes its complete oxidation, with of the mixing solutions extraction of normal hexane-alcohol-water in 4: 1: 1 ratio preparation, get phase, concentrate, get refined volatile oil.Volatile oil after refining separates in the chromatography column of Φ 1.0 * 40cm with 60-100 purpose silica gel, at first adopting the petroleum ether-ethyl acetate mixed solvent is that 300: 1, flow velocity are removed impurity peaks during for 5ml/min in gradient, be that 50: 1, flow velocity are carried out wash-out during for 1ml/min with the petroleum ether-ethyl acetate mixed solvent in gradient again, collect corresponding flow point, concentrate, reclaim solvent, get final product colourless acicular crystal atractylenolide I, about 1.5% (is benchmark with the volatile oil applied sample amount) of yield, purity about 93.9%.
Embodiment 5:
Bighead atractylodes rhizome rhizome is dried naturally, be ground into bighead atractylodes rhizome meal, 2 (60 ℃ the waters bath with thermostatic control of mixed solvent leaching that are made in 40: 1 ratio with the petroleum ether-ethyl acetate of 5 times of volumes, mixing speed 500r/min, each 1h), place after for some time makes its complete oxidation, with normal hexane-ethyl acetate, alcohol and water in 8: 1: 1: the mixing solutions extraction of 1 ratio preparation 3 times, get phase, concentrate, get refined volatile oil.Volatile oil after refining separates in the chromatography column of Φ 1.0 * 40cm with 60-100 purpose silica gel, at first adopting the petroleum ether-ethyl acetate mixed solvent is that 300: 1, flow velocity are removed impurity peaks during for 5ml/min in gradient, be that 50: 1, flow velocity are carried out wash-out during for 1ml/min with the petroleum ether-ethyl acetate mixed solvent in gradient again, collect corresponding flow point, concentrate, reclaim solvent, get final product colourless acicular crystal atractylenolide I, about 1.8% (is benchmark with the volatile oil applied sample amount) of yield, purity about 94.8%.
Gained atractylenolide I crystal is differentiated with gas chromatography-mass spectrography method (GC-MS):
The atractylenolide I crystal that takes a morsel carries out GC-MS and analyzes instrument: TRACE GC 2000/TRACEMS (the thermoelectric instrument company of ThermoQuest).GC condition: chromatographic column DB-WAX30m * 0.25mm * 0.25 μ m, 50 ℃ of (2min)-15 of column temperature ℃/min-260 ℃ (15min), 270 ℃ of sample introduction temperature, splitting ratio 50: 1, carrier gas He, flow velocity 1ml/min.EIMS condition: electron energy 70Ev, mass range 35-450,200 ℃ of ion source temperatures, 250 ℃ of interface temperatures.The retention time of atractylenolide I is 16.23min.
Claims (4)
1. the method for a separating effective ingredient atractylenolide I from Rhizoma Atractylodis Macrocephalae volatile oil is characterized in that, comprises the steps:
(1) exsiccant bighead atractylodes rhizome rhizome is pulverized, powder with in, the solvent leaching of low isopolarity, concentrate, tawny volatile oil crude extract.
(2) the volatile oil crude extract of gained is placed for some time, treat its exhaustive oxidation after, with the mixed extractant solvent of normal hexane-ethyl acetate, alcohol and water, get phase by 4: 0～1: 1～4: 1 arbitrary proportion preparation, concentrate, get purified volatile oil.
(3) purified volatile oil separates with the column chromatography chromatogram method, again with petroleum ether-ethyl acetate by 40～150: the mixed solvent of 1 arbitrary proportion preparation carries out gradient elution, collects corresponding flow point.
(4) concentrate, reclaim solvent, get final product colourless acicular crystal atractylenolide I.
2. according to claim 1 from Rhizoma Atractylodis Macrocephalae volatile oil the method for separating effective ingredient atractylenolide I, it is characterized in that, in the described step (1), described powder leaches with the mixed solvent of petroleum ether-ethyl acetate by 0～40: 1 arbitrary proportion preparation, concentrate with air distillation or distillation under vacuum, get tawny volatile oil crude extract.
3. according to claim 2 from Rhizoma Atractylodis Macrocephalae volatile oil the method for separating effective ingredient atractylenolide I, it is characterized in that described leaching method is that ultrasonic method, cold soaking, heat are carried, diacolation or Soxhlet are extracted.
4. according to claim 1 from Rhizoma Atractylodis Macrocephalae volatile oil the method for separating effective ingredient atractylenolide I, it is characterized in that, in the described step (2), on having got mutually after, use again to wash mutually down and go up phase, concentrate, get purified volatile oil with air distillation or distillation under vacuum.
Priority Applications (1)
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103396890A (en) * | 2013-07-26 | 2013-11-20 | 山西省肿瘤医院 | Method for preparing stable bighead atractylodes rhizome volatile oil |
| CN104490868A (en) * | 2014-12-09 | 2015-04-08 | 上海交通大学医学院附属第三人民医院 | Application of atractylenolide-II derivative in preparation of platelet aggregation resisting drug and platelet aggregation resisting drug |
| CN107556275A (en) * | 2017-10-20 | 2018-01-09 | 南京盖斯夫医药科技有限公司 | A kind of preparation method of atractylenolide |
| CN107674053A (en) * | 2017-10-23 | 2018-02-09 | 南京盖斯夫医药科技有限公司 | A kind of atractylodes lactone V, preparation method and the application in terms of CIK cell culture |
| CN111012815A (en) * | 2020-01-03 | 2020-04-17 | 吉林大学 | Application of bighead atractylodes rhizome volatile oil in preparation of medicine for treating acute liver injury |
-
2008
- 2008-04-22 CN CN2008100607964A patent/CN101265247B/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103396890A (en) * | 2013-07-26 | 2013-11-20 | 山西省肿瘤医院 | Method for preparing stable bighead atractylodes rhizome volatile oil |
| CN103396890B (en) * | 2013-07-26 | 2014-11-05 | 山西省肿瘤医院 | Method for preparing stable bighead atractylodes rhizome volatile oil |
| CN104490868A (en) * | 2014-12-09 | 2015-04-08 | 上海交通大学医学院附属第三人民医院 | Application of atractylenolide-II derivative in preparation of platelet aggregation resisting drug and platelet aggregation resisting drug |
| CN107556275A (en) * | 2017-10-20 | 2018-01-09 | 南京盖斯夫医药科技有限公司 | A kind of preparation method of atractylenolide |
| CN107674053A (en) * | 2017-10-23 | 2018-02-09 | 南京盖斯夫医药科技有限公司 | A kind of atractylodes lactone V, preparation method and the application in terms of CIK cell culture |
| CN107674053B (en) * | 2017-10-23 | 2019-08-20 | 北京恒生佳合细胞科技有限公司 | A kind of atractylodes lactone V, preparation method and the application of aspect is frozen in CIK cell |
| CN111012815A (en) * | 2020-01-03 | 2020-04-17 | 吉林大学 | Application of bighead atractylodes rhizome volatile oil in preparation of medicine for treating acute liver injury |
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| CN101265247B (en) | 2011-05-04 |
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