CN101709028A - Method for extracting and separating curdione from oil of zedoary turmeric - Google Patents
Method for extracting and separating curdione from oil of zedoary turmeric Download PDFInfo
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- CN101709028A CN101709028A CN200910177025A CN200910177025A CN101709028A CN 101709028 A CN101709028 A CN 101709028A CN 200910177025 A CN200910177025 A CN 200910177025A CN 200910177025 A CN200910177025 A CN 200910177025A CN 101709028 A CN101709028 A CN 101709028A
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Abstract
The invention relates to a method for extracting and purifying curdione from the oil of zedoary turmeric, a natural medicinal plant volatile oil. The purity of the curdione obtained by the method reaches 99.9%, and the curdione conforms to the requirement of the purity of a contrast product in the field of traditional Chinese medicine and provides a superior raw material for developing a subsequent product. The method comprises the following steps: mixing the oil of the zedoary turmeric and silica gel used for chromatography evenly, coating a silica gel column, taking petroleum ether and ethyl acetate which have different proportions as an eluent for gradient elution, collecting and merging eluant containing the component of the curdione, concentrating, refrigerating, standing, precipitating crystals and separating and purifying the crystals again by the same method to obtain recrystallized crystals so as to obtain the curdione. The method features simple operation and good repeatability.
Description
Technical field
The present invention relates to a kind of extracting method that extracts single active ingredient from natural medicinal plant volatile oil, be specifically related to a kind ofly to extract from zedoary turmeric oil, the method for purifying curdione, the curdione purity of this method preparation is up to 99.9%.
Background technology
Zedoary turmeric oil is the volatile oil that extracts from multiple medicinal plant, as all containing zedoary turmeric oil in the zingiberaceous plants such as turmeric, curcuma zedoary, root tuber of aromatic turmeric.Contain the various active composition in the zedoary turmeric oil, have promoting flow of QI and blood, the long-pending pain relieving that disappears, promoting blood circulation and removing blood stasis, remove necrosis and promote granulation, the effect of enhancing body immunological competence.Modern pharmacology studies show that zedoary turmeric oil directly suppresses, destruction of cancer cells; Anti-HPV infects; Zedoary turmeric oil has collaborative killing action to pathogenic micro-organisms such as bacterium, mould, trichomonad, viruses, and repairs pathological tissues, promotion wound healing.Aromatic turmeric oil preparation is used for the treatment of vaginal infection, colpitis mycotica, trichomonal vaginitis, cervical erosion and the treatment viral infection that Candida albicans causes; Also be used for prevention, treatment cervical cancer.Its anticancer, antibiotic, antiviral main active ingredient is Curcumenol, curdione, Elemenum, germacrone etc., having at present a lot clinically is the preparation of effective constituent with the zedoary turmeric oil, as: Zedoary turmeric oil glucose injection, Oleum Curcumae injection, Compound Zedoary Turmeric Oil Suppositories, compound zedoary oil soft capsule, Baofukang suppositories etc., be widely used in treatment and prophylaxis of viral infections, the cervical cancer pathology of the gynaecopathia of bacterium, mould, candida albicans infection and HPV repeated infection.In the past, because the Zedoary turmeric oil complexity, active constituent content is lower, and analytical procedure is limited, has limited the quality control of preparation effective constituent.Current, along with the progress and the development of analytical instrument, analytical procedure, can detect the monomeric substance of zedoary turmeric oil effective constituent more exactly, for aromatic turmeric oil preparation accurately composition qualitative, quantitatively provide condition.Curdione is a main active ingredient in the zedoary turmeric oil, account for about 15% of oil of zedoary turmeric contents, the present application people passes through test repeatedly, the effective ways of extraction, purifying curdione from zedoary turmeric oil have been found, can be for the aromatic turmeric oil preparation composition detection provide highly purified reference substance, and provide the fine raw material for the exploitation of curdione subsequent product.
Extract the curdione method in the prior art and normally adopt the sherwood oil wash-out earlier, use sherwood oil again: ethyl acetate (9: 1) wash-out, but this single type of elution is difficult to remove effectively impurity.In view of complicated component in the zedoary turmeric oil, the application is with sherwood oil (60～90 ℃): the ethyl acetate volume ratio is 100: 0,30: 1,25: 1,20: 1,15: 1,12: 1, carry out gradient elution at 9: 1, and with sherwood oil (60-90 ℃): the ethyl acetate volume ratio is 18: 1,15: 1,12: 1 elutriant carries out the secondary wash-out, wash-out by secondary, a plurality of gradients is removed most of impurity, and to guarantee the purity of curdione, the curdione purity that the application obtains is up to 99.9%.The described method of the application and other extracting and purifying method compare, and it is easy also to have experimental implementation, and equipment requirements is simple, and condition reaches easily, and yield is big, the characteristics that purity is high.
Summary of the invention
The object of the invention provide a kind of monomer of highly purified curdione and from zedoary turmeric oil, extract, the method for purifying curdione, monomer purity has solved the problem that always lacks high purity curdione reference substance in the prior art up to 99.9%.
Extraction of the present invention, purification process are as follows:
A certain amount of zedoary turmeric oil and an amount of column chromatography are mixed thoroughly with silica gel, on the silicagel column that is pre-charged with, carry out gradient elution with sherwood oil (60～90 ℃) and ethyl acetate mixed solvent successively, sherwood oil wherein: the ethyl acetate volume ratio is followed successively by: 100: 0,30: 1,25: 1,20: 1,15: 1,12: 1,9: 1, collect the elutriant under each gradient, every part of 30ml, immediately many parts of elutriants under each gradient are detected with thin-layer method, at any time monitor the wash-out situation of curdione, when wash-out proceeds to sherwood oil: when the ethyl acetate volume ratio was 12: 1 and 9: 1, curdione was come out by wash-out successively, wherein concentrated under 9: 1 gradient, detected result under the gradient of 12: 1 and 9: 1 is shown that many parts of elutriants that contain single curdione spot merge, and concentrate 0～2 ℃ of refrigeration, placed 24～48 hours, there is crystal to separate out, filters, stand-by.Get chromatography column and add column chromatography with silica gel (105 ℃ drying 6 hours), tamp, vacuum pump was taken out 6 hours, and with sherwood oil (60～90 ℃): ethyl acetate (18: 1) is walked post.Get crystal sherwood oil (60～90 ℃) dissolving that the step obtains, mix thoroughly with an amount of silica gel for chromatography, last silicagel column, adjust the ratio of elutriant, successively with sherwood oil (60-90 ℃): the ethyl acetate volume ratio is 18: 1,15: 1, with method carry out gradient elution at 12: 1, collect the elutriant under each gradient, every part of 20ml detects with thin-layer method many parts of elutriants under each gradient immediately, and detected result under 12: 1 gradients is shown that many parts of elutriants that contain single curdione spot merge, concentrate, 0～2 ℃ of refrigeration, place crystallization in 24～48 hours, crystallisate carries out recrystallization with appropriate organic solvent, promptly gets highly purified curdione.
In the aforesaid method, silicagel column is preferably the silicagel column of dry-packing; Recrystallization solvent is selected from sherwood oil, propyl carbinol or normal hexane, is preferably normal hexane.
Description of drawings
Accompanying drawing 1: the gas-matter color atlas of gas chromatograph-mass spectrometer analysis self-control curdione
Accompanying drawing 2: the analysis report of high performance liquid chromatography quantitative analysis self-control curdione and commercially available product
Embodiment
Getting chromatography column adds column chromatography and fills out post with silica gel (105 ℃ drying 6 hours) 300g dry method, tamp, vacuum pump was taken out 6 hours, and with sherwood oil (60～90 ℃): ethyl acetate (18: 1) is walked post, gets zedoary turmeric oil 30g, strutting silica gel for chromatography 15g mixes thoroughly, the dry method upper prop is used linear gradient elution method, carries out gradient elution with sherwood oil (60～90 ℃) and ethyl acetate mixed solvent successively, sherwood oil wherein: the ethyl acetate volume ratio was followed successively by 100: 0,30: 1,25: 1,20: 1,15: 1,12: 1,9: 1, collect the elutriant under each gradient, every part of 30ml, immediately many parts of elutriants under each gradient are carried out the thin layer inspection, the wash-out situation of monitoring curdione shows that with detected result under the gradient of 12: 1 and 9: 1 many parts of elutriants that contain single curdione spot merge, concentrate, 0～2 ℃ of refrigeration was placed 48 hours, had crystal to separate out, with sintered filter funnel leaching crystallization, stand-by.Get chromatography column and add column chromatography with silica gel (105 ℃ drying 6 hours) 30g, dry method is filled out post, tamps, and vacuum pump was taken out 6 hours, and with sherwood oil (60～90 ℃): ethyl acetate (18: 1) is walked post.Get crystal sherwood oil (60～90 ℃) dissolving that the step obtains, mix thoroughly with an amount of column chromatography silica gel, be leather hard, last silicagel column, carry out gradient elution, wherein sherwood oil with method with sherwood oil (60～90 ℃) and ethyl acetate mixed solvent: the ethyl acetate volume ratio was followed successively by 18: 1,15: 1,12: 1, collect elutriant, every part of 20ml immediately carries out the thin layer inspection to many parts of elutriants under each gradient, at any time monitor the wash-out situation of curdione, detected result under 12: 1 gradients is shown that many parts of elutriants that contain single curdione spot merge, and concentrate 0～2 ℃ of refrigeration, placed 48 hours, there is crystal to separate out, uses the sintered filter funnel leaching, with the crystal that obtains normal hexane recrystallization, promptly get highly purified curdione (1.5g), purity is 99.9%.
Get chromatography column and add column chromatography with silica gel (105 ℃ drying 6 hours) 300g, dry method is filled out post, tamps, and vacuum pump was taken out 6 hours, with sherwood oil (60～90 ℃): ethyl acetate (18: 1) is walked post, get zedoary turmeric oil 30g, strutting silica gel for chromatography 15g mixes thoroughly, the dry method upper prop, use linear gradient elution method, carry out gradient elution, wherein sherwood oil with sherwood oil (60～90 ℃) and ethyl acetate mixed solvent successively: the ethyl acetate volume ratio was followed successively by 100: 0,30: 1,25: 1,20: 1,15: 1,12: 1,9: 1, collect the elutriant under each gradient, every part of 30ml immediately carries out the thin layer inspection to many parts of elutriants under each gradient, the wash-out situation of monitoring curdione, detected result under the gradient of 12: 1 and 9: 1 is shown that many parts of elutriants that contain single curdione spot merge, and concentrate 0～2 ℃ of refrigeration, placed 48 hours, there is crystal to separate out, with sintered filter funnel leaching crystallization, stand-by.Get chromatography column and add column chromatography with silica gel (105 ℃ drying 6 hours) 30g, dry method is filled out post, tamps, and vacuum pump was taken out 6 hours, and with sherwood oil (60～90 ℃): ethyl acetate (18: 1) is walked post.Get crystal sherwood oil (60～90 ℃) dissolving that the step obtains, mix thoroughly with an amount of column chromatography silica gel, be leather hard, last silicagel column, carry out gradient elution, wherein sherwood oil with method with sherwood oil (60～90 ℃) and ethyl acetate mixed solvent: the ethyl acetate volume ratio was followed successively by 18: 1,15: 1,12: 1, collect elutriant, every part of 20ml immediately carries out the thin layer inspection to many parts of elutriants under each gradient, at any time monitor the wash-out situation of curdione, detected result under 12: 1 gradients is shown that many parts of elutriants that contain single curdione spot merge, and concentrate 0～2 ℃ of refrigeration, placed 24 hours, there is crystal to separate out, uses the sintered filter funnel leaching, with the crystal that obtains normal hexane recrystallization, promptly get highly purified curdione (1.2g), purity is 99.7%.
Get chromatography column and add column chromatography with silica gel (105 ℃ drying 6 hours) 600g, dry method is filled out post, tamps, and vacuum pump was taken out 6 hours, with sherwood oil (60～90 ℃): ethyl acetate (18: 1) ethyl acetate is walked post, get zedoary turmeric oil 60g, strutting silica gel for chromatography 30g mixes thoroughly, the dry method upper prop, use linear gradient elution method, carry out gradient elution, wherein sherwood oil with sherwood oil (60～90 ℃) and ethyl acetate mixed solvent successively: the ethyl acetate volume ratio was followed successively by 100: 0,30: 1,25: 1,20: 1,15: 1,12: 1,9: 1, collect the elutriant under each gradient, every part of 30ml immediately carries out the thin layer inspection to many parts of elutriants under each gradient, the wash-out situation of monitoring curdione, detected result under the gradient of 12: 1 and 9: 1 is shown that many parts of elutriants that contain single curdione spot merge, and concentrate 0～2 ℃ of refrigeration, placed 48 hours, there is crystal to separate out, with sintered filter funnel leaching crystallization, stand-by.Get chromatography column and add column chromatography with silica gel (105 ℃ drying 6 hours) 60g, dry method is filled out post, tamps, and vacuum pump was taken out 6 hours, and with sherwood oil (60～90 ℃): ethyl acetate (18: 1) is walked post.Get crystal sherwood oil (60～90 ℃) dissolving that the step obtains, mix thoroughly with an amount of column chromatography silica gel, be leather hard, last silicagel column, carry out gradient elution, wherein sherwood oil with method with sherwood oil (60～90 ℃) and ethyl acetate mixed solvent: the ethyl acetate volume ratio was followed successively by 18: 1,15: 1,12: 1, collect elutriant, every part of 20ml immediately carries out the thin layer inspection to many parts of elutriants under each gradient, at any time monitor the wash-out situation of curdione, detected result under 12: 1 gradients is shown that many parts of elutriants that contain single curdione spot merge, and concentrate 0～2 ℃ of refrigeration, placed 24 hours, there is crystal to separate out, with the sintered filter funnel leaching of this crystal, with the crystal that obtains normal hexane recrystallization, promptly get highly purified curdione (3.2g), purity is 99.8%.
Embodiment 4
The curdione that embodiment 1 is obtained carries out qualitative identification and analysis with gas chromatograph-mass spectrometer, experiment condition: GC conditions: quartz capillary column HP-FFAP (30m * 0.25mm, 0.25um), temperature programming: since 160 ℃, keep 20min, be warmed up to 250 ℃ with 20 ℃/min again, carrier gas is He, post flow 3.0ml/min, 250 ℃ of injector temperatures, splitting ratio 50: 1.
Mass spectrum condition: EI source; Ionization voltage 70eV, 230 ℃ of ion source temperatures, sweep limit 40～500aum, sample size 1.0uL.
Result such as accompanying drawing 1 show: self-control curdione mass spectrum cracking pattern and computer mass-spectrometric data storehouse curdione collection of illustrative plates are in full accord, and the material that obtains with the inventive method really is curdione.
Curdione and commercially available curdione that embodiment 1 is obtained carry out the quantitative comparison analysis with high performance liquid chromatograph, experiment condition: liquid phase chromatogram condition: octadecylsilane chemically bonded silica is a weighting agent; With acetonitrile-0.1% acetic acid aqueous solution (40: 60) is moving phase; The detection wavelength is 230nm; Flow velocity is 1ml/min.
Result such as accompanying drawing 2, this result shows: the content of commercially available curdione only is 98.1%, the curdione content that extracts purifying with the inventive method is 99.9%.
The analytical results of commercially available curdione
The curdione analytical results of the embodiment of the present application 1
The present invention is by optimizing chromatographic condition and crystallization condition, from zedoary turmeric oil, obtained highly purified curdione monomer, for the detection by quantitative of the product that contains zedoary turmeric oil provides highly purified reference substance, also provide the fine raw material for other Products Development that contain curdione.Above embodiment only exemplifies, and is not limitation of the present invention, and other embodiment or equivalents of carrying out simple transformation on basis of the present invention also will fall into protection scope of the present invention.
Claims (5)
1. the method for extraction, purifying curdione from a zedoary turmeric oil comprises the steps:
Zedoary turmeric oil and an amount of column chromatography are mixed thoroughly with silica gel, on the silicagel column filled in advance, carry out gradient elution with 60～90 ℃ sherwood oils and ethyl acetate mixed solvent, sherwood oil wherein: the ethyl acetate volume ratio is followed successively by: 100: 0,30: 1,25: 1,20: 1,15: 1,12: 1,9: 1, collect the elutriant under each gradient, every part of 30ml, immediately many parts of elutriants under each gradient are detected with thin-layer method, detected result under 12: 1 and 9: 1 gradients is shown that many parts of elutriants that contain single curdione spot merge, concentrate, 0～2 ℃ of refrigeration was placed 24～48 hours, had crystal to separate out, filter, crystal is mixed the silicagel column of upward filling in advance with sherwood oil (60～90 ℃) dissolving, solution thoroughly with an amount of column chromatography with silica gel, carry out gradient elution with 60～90 ℃ sherwood oil and ethyl acetate mixed solvent once more, adjust the ratio of elutriant, wherein sherwood oil: the ethyl acetate volume ratio was followed successively by 18: 1,15: 1,12: 1, collect the elutriant under each gradient, every part of 20ml detects with thin-layer method many parts of elutriants under each gradient immediately, detected result under 12: 1 gradients is shown that many parts of elutriants that contain single curdione spot merge, concentrate, 0～2 ℃ of refrigeration, place crystallization in 24～48 hours, crystallisate carries out recrystallization with appropriate organic solvent, promptly gets the curdione product.
2. the method for claim 1, wherein said silicagel column is the silicagel column of dry-packing.
3. method as claimed in claim 1 or 2, wherein said recrystallization solvent is selected from sherwood oil, propyl carbinol or normal hexane.
4. method as claimed in claim 3, recrystallization solvent wherein is preferably normal hexane.
5. claim 1 or 2 described methods, wherein the purity of curdione is 99.8%-99.9%.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102091059A (en) * | 2010-12-24 | 2011-06-15 | 海南碧凯药业有限公司 | Application of curdione |
| CN102716112A (en) * | 2012-07-13 | 2012-10-10 | 海南碧凯药业有限公司 | Pharmaceutical composition capable of resisting HPV (human papillomavirus) infection |
| CN105688028A (en) * | 2014-11-28 | 2016-06-22 | 天津耀宇生物技术有限公司 | A method of extracting anti-lung-cancer active components from zedoray rhizome and applications of the active components |
| CN105738546A (en) * | 2014-12-12 | 2016-07-06 | 桂林八加一药物研究股份有限公司 | Establishment method of curcuma aromatica medicine fingerprint map and the fingerprint map thereof |
| CN109876115A (en) * | 2019-04-16 | 2019-06-14 | 石药集团远大(大连)制药有限公司 | It is a kind of to treat skin trauma, the composition of infectious disease and preparation method and application |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1284534C (en) * | 2004-01-13 | 2006-11-15 | 李绍平 | Aromatic turmeric oil extracts and its preparing process and medicinal composition containing the same |
| CN101948456B (en) * | 2010-07-29 | 2015-09-09 | 沈阳药科大学 | Class Curcumalactone derivative with anti-tumor activity and preparation method thereof |
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2009
- 2009-09-17 CN CN2009101770258A patent/CN101709028B/en active Active
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102091059A (en) * | 2010-12-24 | 2011-06-15 | 海南碧凯药业有限公司 | Application of curdione |
| CN102716112A (en) * | 2012-07-13 | 2012-10-10 | 海南碧凯药业有限公司 | Pharmaceutical composition capable of resisting HPV (human papillomavirus) infection |
| CN102716112B (en) * | 2012-07-13 | 2014-05-14 | 海南碧凯药业有限公司 | Pharmaceutical composition capable of resisting HPV (human papillomavirus) infection |
| CN105688028A (en) * | 2014-11-28 | 2016-06-22 | 天津耀宇生物技术有限公司 | A method of extracting anti-lung-cancer active components from zedoray rhizome and applications of the active components |
| CN105738546A (en) * | 2014-12-12 | 2016-07-06 | 桂林八加一药物研究股份有限公司 | Establishment method of curcuma aromatica medicine fingerprint map and the fingerprint map thereof |
| CN109876115A (en) * | 2019-04-16 | 2019-06-14 | 石药集团远大(大连)制药有限公司 | It is a kind of to treat skin trauma, the composition of infectious disease and preparation method and application |
| CN109876115B (en) * | 2019-04-16 | 2021-07-30 | 石药集团远大(大连)制药有限公司 | Composition for treating skin wound and infection diseases, preparation method and application |
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