CN107383148A - Purification shionon technique is extracted from beautiful millettia root - Google Patents
Purification shionon technique is extracted from beautiful millettia root Download PDFInfo
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- CN107383148A CN107383148A CN201710528025.2A CN201710528025A CN107383148A CN 107383148 A CN107383148 A CN 107383148A CN 201710528025 A CN201710528025 A CN 201710528025A CN 107383148 A CN107383148 A CN 107383148A
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- shionon
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- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J63/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by expansion of only one ring by one or two atoms
- C07J63/008—Expansion of ring D by one atom, e.g. D homo steroids
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Abstract
The invention discloses purification shionon technique is extracted from beautiful millettia root, belong to technical field of natural product extraction, including the step such as beautiful millettia root drying, crushing, coarse extraction, concentration, resin extraction, refined.Described extracts purification shionon technique from beautiful millettia root, and each step is concise, it is easy to accomplish large-scale production, extracts gained shionon content height, impurity content is low.
Description
Technical field
The invention belongs to field of natural product extraction, and in particular to the method for extraction purification shionon from beautiful millettia root.
Background technology
Beautiful millettia root, alias pig's feet large bamboo hat with a conical crown and broad brim, Jin Zhonggen, beautiful millettia root, hang upside down Jin Zhong, energetically potato.Planted for pulse family pulse family Millettia
Thing, beautiful millettia Millettia specisoa Champ..Beautiful millettia root is used as medicine with root, and Main Ingredients and Appearance is protein, starchiness
And alkaloid etc..Beautiful millettia root has very high medical value, early in《The sward property of medicine is standby will》In recorded that " strong muscle is active, qi-restoratives
Moistening lung, control pain in waist and lower extremities, arthralgia pain due to rheumatism, chronic hepatitis, pulmonary tuberculosis ".《Luchuan book on Chinese herbal medicine》Also it is recorded：" removing heat from the lung to relieve cough, refrigerant solution
Poison.Hemoptysis, dysentery are controlled, warm disease body heat is thirsty, dizzy ".Existing pharmacological research shows that beautiful millettia root has immunological regulation, drop blood
Sugared, anti-oxidant and removing free radical and other effects, contains the chemical constituents such as triterpene, flavones, phenols, lignin in beautiful millettia root root tuber,
By effective chemical constituent separating-purifying in beautiful millettia root, pharmacology and nutrient for studying beautiful millettia root are significant.
Shionon is a kind of triterpenes components in aster rhizome, and its chemical formula is C30H50O.Shionon is colourless crystallization
Powder, the organic solvents such as petroleum ether, benzene, ether, chloroform can be dissolved in, it is not soluble in water.Aster generally carries from the root of plant aster
Take, but in some other plant content also compared with horn of plenty.Shionon has the effect of moistening lung to lower qi, stopping coughing and removing phlegm, is usually used in
Addition is in some relieving cough and reducing sputum medicines.In modern medicine study, the characteristic effect of shionon is also demonstrate that, it is such as general to going
The inhibitory action of the activity of elementization enzyme 2,1L-1 β, TNF-α and NO influence etc. are discharged to lipopolysaccharide-induced macrophage.
In recent years, as the continuous expansion of beautiful millettia root cultivated area, the continuous growth of yield, beautiful millettia root are eaten as a kind of medicine
Homologous Chinese medicine, the theoretical research for its property of medicine are more and more.Such as paper《Beautiful millettia root petroleum ether part chemical composition is ground
Study carefully》It is the structure for identifying different compounds after being isolated and purified to 95% ethanol extract of beautiful millettia root by spectrographic technique,
Confirm in beautiful millettia root containing shionon, coffee acid rain fan beans alcohol ester, dipeptides Aurantiamide acetate etc.；Application for a patent for invention document
《A kind of Millettia speciosa Champ.extract compound tea and preparation method thereof》One kind is then disclosed by Millettia speciosa Champ.extract, Fructus Monordicae extract etc.
The compound tea of composition, it is disclosed that the extracting method of beautiful millettia root, including continuous flow upstream ultrasonic extraction, centrifugation, enzymolysis, ultrafiltration etc.
The Millettia speciosa Champ.extract that step is prepared.But in existing document, rarely have for the Flavonoid substances in beautiful millettia root, especially
It is that the research of shionon extraction is recorded.
The disclosure of background above technology contents is only used for inventive concept and the technical scheme that auxiliary understands the present invention, and it is not
The prior art of present patent application is necessarily belonged to, shows the applying date of the above in present patent application in no tangible proof
In the case of disclosed, above-mentioned background technology should not be taken to evaluate the novelty and creativeness of the application.
The content of the invention
Purification shionon technique is extracted from beautiful millettia root the technical problem to be solved in the present invention is to provide one kind, to realize from ox
Scale, industrialization extraction shionon composition energetically, controllability is strong, efficiency high.
In order to solve the above technical problems, the present invention uses following technical scheme：
Purification shionon technique is extracted from beautiful millettia root, is comprised the following steps：
a1：Pre-treatment：After the cleaned outer layer impurity of fresh beautiful millettia root, the section that thickness is 0.5~1.0cm is cut into, is divided
Power is positioned over to dry 1~2h under 500~800w infrared lamps, then is placed in hot air drier and dries；
a2：Coarse extraction：By beautiful millettia root obtained by step a1 after being crushed to 50~80 mesh, it is placed in 95% ethanol solution,
Extraction, filtering are heated at 60~85 DEG C, filter residue repeats extraction 2 times, merges extract solution；
a3：It is concentrated and dried：Molten extract solution obtained by step a2 through being evaporated under reduced pressure, recycling design, is obtained into crude extract；
a4：Extraction：By crude extract deionized water dissolving obtained by step a3, extracted 2 times with petroleum ether normal temperature, merge stone
Oily ether moiety；
a5：Column chromatography for separation is extracted：Step a4 resulting solutions are crossed into polarity silicagel column, use 60% petroleum ether and 40% second
Acetoacetic ester mixed liquor elutes, and obtains eluent, is evaporated under reduced pressure and removes solvent；
a6：Resin extracts：By extract obtained 85% ethanol in mass ratio 1 of step a4:1 stirring and dissolving, solution warp
D101B types macroporous absorbent resin adsorbs 2 times；
a7：Elution：Use concentration for 10%~85% ethanol solution, as low concentration in high concentration to described in step a6
Resin gradient elution, obtains eluent；
a8：Filtering：By eluent obtained by step a7 through the milipore filter that molecular cut off is 1000, filtrate is collected；
a9：Concentration：Ultrafiltrate decompression obtained by step a5 is boiled off into solvent；
a10：Dry：Step a7 gains are placed in vacuum drying chamber, 1.5~3h is dried at 60~80 DEG C.
Preferably, it is described to extract purification shionon technique from beautiful millettia root, dry in step a1 to moisture be 8~
15%；
Preferably, described to extract purification shionon technique from beautiful millettia root, grinding mode described in step a2 is hammer leaf powder
Broken machine, fluid energy mill, one kind of ball mill；
Preferably, described to extract purification shionon technique from beautiful millettia root, it is rope that leaching mode is heated described in step a2
One kind in formula extraction method, decocting method；
Preferably, described to extract purification shionon technique from beautiful millettia root, the vapo(u)rizing temperature being evaporated under reduced pressure in step a3 is
65~75 DEG C, distillation pressure is 50~150Pa.
Preferably, described extracted from beautiful millettia root purifies shionon technique, and the petroleum ether boiling range used in step a4 is
60~90 DEG C.
Preferably, described to extract purification shionon technique from beautiful millettia root, the large aperture resin described in step a6 is 50
~100nm apertures, the D101B type macroporous absorbent resins of 35%~60% cell size.
Preferably, described extracted from beautiful millettia root purifies shionon technique, and in step a6, resin adsorption process keeps solution
Temperature is 25~35 DEG C, and adsorption flow rate is 2~5BV/h.
Preferably, described to extract purification shionon technique from beautiful millettia root, ultrafiltration membrane material described in step a8 is poly- third
Alkene nitrile, polyether sulfone, Kynoar.
It is highly preferred that described extract purification shionon technique from beautiful millettia root, in step a8 filter pressure be 0.1~
0.15MPa。
The invention has the advantages that：
(1) in the extraction process of shionon of the present invention, beautiful millettia root fresh goods is first dehydrated through infra-red drying, is effectively killed
Bacterium, mould；
(2) in the extraction process of shionon of the present invention, beautiful millettia root is respectively through column chromatography, macroporous absorbent resin, super
Filter membrane extracts, polysaccharide and other major impurity good separating effects contained by beautiful millettia root；
(3) extraction of shionon is carried out using technique of the present invention, the shionon rate of recovery of institute's output is high, and purity is good,
Be advantageous to the further research of shionon content and property in beautiful millettia root.
Embodiment
The present invention is described in further detail with reference to embodiment.It is emphasized that the description below is only
Only it is exemplary, the scope being not intended to be limiting of the invention and its application.
Embodiment 1：
Purification shionon technique is extracted from beautiful millettia root, is comprised the following steps：
a1：Pre-treatment：After the cleaned outer layer impurity of fresh beautiful millettia root, the section that thickness is 0.5cm is cut into, divides and is positioned over
Power is to dry 2h under 500w infrared lamps, then it is 8% to be placed in drying to moisture in hot air drier；
a2：Coarse extraction：By beautiful millettia root obtained by step a1 after hammer crusher is crushed to 50 mesh, 95% ethanol solution is placed in
In, extraction 2 times is repeated, merges extract solution using the heating extraction of soxhlet extraction method, filtering, filter residue at 60 DEG C；
a3：It is concentrated and dried：In vapo(u)rizing temperature it is 65 DEG C by molten extract solution obtained by step a2, distillation pressure is to be depressurized under 50Pa
Distillation, recycling design, obtains crude extract；
a4：Extraction：It is normal with the petroleum ether that boiling range is 60~90 DEG C by crude extract deionized water dissolving obtained by step a3
Temperature extraction 2 times, merges oil ether moiety；
a5：Column chromatography for separation is extracted：Step a4 resulting solutions are crossed into polarity silicagel column, use 60% petroleum ether and 40% second
Acetoacetic ester mixed liquor elutes, and obtains eluent, is evaporated under reduced pressure and removes solvent；
a6：Resin extracts：By extract obtained 85% ethanol in mass ratio 1 of step a4:1 stirring and dissolving, solution warp
50nm apertures, the D101B types macroporous absorbent resin of 35% cell size adsorb 2 times, and it is 25 DEG C that solution temperature is kept in adsorption process, is inhaled
Attached flow velocity is 2BV/h；
a7：Elution：Use concentration for 10%~85% ethanol solution, as low concentration in high concentration to described in step a6
Resin gradient elution, obtains eluent；
a8：Filtering：By eluent obtained by step a7 in the case where filter pressure is 0.1MPa, through molecular cut off be 1000 it is poly-
Acrylonitrile milipore filter, collect filtrate；
a9：Concentration：Ultrafiltrate decompression obtained by step a5 is boiled off into solvent；
a10：Dry：Step a7 gains are placed in vacuum drying chamber, 3h is dried at 60 DEG C.
Embodiment 2
Purification shionon technique is extracted from beautiful millettia root, is comprised the following steps：
a1：Pre-treatment：After the cleaned outer layer impurity of fresh beautiful millettia root, the section that thickness is 1.0cm is cut into, divides and is positioned over
Power is to dry 1h under 800w infrared lamps, then it is 15% to be placed in drying to moisture in hot air drier；
a2：Coarse extraction：By beautiful millettia root obtained by step a1 after fluid energy mill is crushed to 80 mesh, it is placed in 95% ethanol solution,
Extraction 2 times is repeated, merges extract solution using decocting method heating extraction, filtering, filter residue at 85 DEG C；
a3：It is concentrated and dried：In vapo(u)rizing temperature it is 75 DEG C by molten extract solution obtained by step a2, distillation pressure is to subtract under 150Pa
Pressure distillation, recycling design, obtains crude extract；
a4：Extraction：It is normal with the petroleum ether that boiling range is 60~90 DEG C by crude extract deionized water dissolving obtained by step a3
Temperature extraction 2 times, merges oil ether moiety；
a5：Column chromatography for separation is extracted：Step a4 resulting solutions are crossed into polarity silicagel column, use 60% petroleum ether and 40% second
Acetoacetic ester mixed liquor elutes, and obtains eluent, is evaporated under reduced pressure and removes solvent；
a6：Resin extracts：By extract obtained 85% ethanol in mass ratio 1 of step a4:1 stirring and dissolving, solution warp
100nm apertures, the D101B types macroporous absorbent resin of 60% cell size adsorb 2 times, and it is 35 DEG C that solution temperature is kept in absorption, absorption
Flow velocity is 5BV/h；
a7：Elution：Use concentration for 10%~85% ethanol solution, as low concentration in high concentration to described in step a6
Resin gradient elution, obtains eluent；
a8：Filtering：By eluent obtained by step a7 in the case where filter pressure is 0.15MPa through molecular cut off be 1000 it is poly-
Ether sulfone milipore filter, collect filtrate；
a9：Concentration：Ultrafiltrate decompression obtained by step a5 is boiled off into solvent；
a10：Dry：Step a7 gains are placed in vacuum drying chamber, 1.5h is dried at 80 DEG C.
Embodiment 3
Purification shionon technique is extracted from beautiful millettia root, is comprised the following steps：
a1：Pre-treatment：After the cleaned outer layer impurity of fresh beautiful millettia root, the section that thickness is 0.8cm is cut into, divides and is positioned over
Power is that 1~2h is dried under 600w infrared lamps, then it is 10% to be placed in drying to moisture in hot air drier；
a2：Coarse extraction：By beautiful millettia root obtained by step a1 after ball mill is crushed to 60 mesh, it is placed in 95% ethanol solution,
Extraction 2 times is repeated, merges extract solution using the heating extraction of soxhlet extraction method, filtering, filter residue at 75 DEG C；
a3：It is concentrated and dried：In vapo(u)rizing temperature it is 70 DEG C by molten extract solution obtained by step a2, distillation pressure is to subtract under 100Pa
Pressure distillation, recycling design, obtains crude extract；
a4：Extraction：It is 60~90 DEG C of petroleum ether normal temperature with boiling range by crude extract deionized water dissolving obtained by step a3
Extraction 2 times, merge oil ether moiety；
a5：Column chromatography for separation is extracted：Step a4 resulting solutions are crossed into polarity silicagel column, use 60% petroleum ether and 40% second
Acetoacetic ester mixed liquor elutes, and obtains eluent, is evaporated under reduced pressure and removes solvent；
a6：Resin extracts：By extract obtained 85% ethanol in mass ratio 1 of step a4:1 stirring and dissolving, solution warp
80nm apertures, the D101B types macroporous absorbent resin of 50% cell size adsorb 2 times, and it is 30 DEG C that solution temperature is kept during absorption, absorption stream
Speed is 4BV/h；
a7：Elution：Use concentration for 10%~85% ethanol solution, as low concentration in high concentration to described in step a6
Resin gradient elution, obtains eluent；
a8：Filtering：It is 1000 through molecular cut off by eluent obtained by step a7 in the case where filter pressure is 0.12MPa
Polyvinylidene fluoride (PVDF) ultrafiltration membrane, collect filtrate；
a9：Concentration：Ultrafiltrate decompression obtained by step a5 is boiled off into solvent；
a10：Dry：Step a7 gains are placed in vacuum drying chamber, 2h is dried at 75 DEG C.
To describe beneficial effects of the present invention in detail, it is further provided experimental result.
By equivalent, be divided into 4 groups of equivalent with place of production fresh beautiful millettia root, every group of 500g, respectively the experimental method through control group with
3 embodiment methods describeds of the present invention carry out the extraction of shionon, weigh extract quality with assay balance, use liquid chromatography
The content of shionon in Detection and Extraction thing.Specific experiment condition and result are as follows.
Control group ox extracts shionon method energetically：By beautiful millettia root fresh goods through 120 DEG C of heated-air dryings to moisture≤
15%th ,≤50 mesh particles are ground into, with after 95% ethanol immersion 24h, heating and refluxing extraction 3 times, merge extract solution, be evaporated under reduced pressure
Ethanol extract is obtained, with the petroleum ether extraction 3 times that boiling range is 60~90 DEG C, combining extraction liquid, is pressed using petroleum ether-ethyl acetate
9:1 to 0:1 gradient elution, polarity silicagel column is crossed to eluent, collect main component, be evaporated under reduced pressure, vacuum dried case is dried.
Shionon detection method of content：Shionon standard items (HPLC >=98%) 0.020g is weighed to be dissolved in 20ml chloroforms,
0.0010g/ml shionon standard liquids are made into, it is standby with 0.45 μm of membrane filtration.Select aster end absorption detecting wavelength for
208nm.Forward chromatographic column, 35 DEG C of column temperature, mobile phase V are used as from Shimadzu Shimadzu ODS (150mm × 4.6mm, 5 μm)
(acetonitrile):V (water)=95:5, flow velocity 1.0ml/min, with high performance liquid chromatograph examination criteria product, and record its peak shape area.
Chromatographic peak is also wanted to using identical inspection method 4 groups of extracts of detection again, it contains with the liquid phase calculated by peak area of 4 groups of extracts
Amount.
From experimental result, compared to the general extraction methods of control group, beautiful millettia root is entered using the method for the invention
The extraction of row shionon, there is the advantage that extract quality is more, shionon content is high.
The specific implementation of the invention is not to be limited to these illustrations for above content, is led for technology belonging to the present invention
For the those of ordinary skill in domain, without departing from the inventive concept of the premise, some simple deduction or replace can also be made,
The scope of patent protection that the present invention is determined by the claims submitted should be all considered as belonging to.
Claims (10)
1. extract purification shionon technique from beautiful millettia root, it is characterised in that comprise the following steps：
a1：Pre-treatment：After the cleaned outer layer impurity of fresh beautiful millettia root, the section that thickness is 0.5~1.0cm is cut into, divides placement
It is that 1~2h is dried under 500~800w infrared lamps in power, then is placed in hot air drier and dries；
a2：Coarse extraction：By beautiful millettia root obtained by step a1 after being crushed to 50~80 mesh, it is placed in 95% ethanol solution, 60~
Extraction, filtering are heated at 85 DEG C, filter residue repeats extraction 2 times, merges extract solution；
a3：It is concentrated and dried：Molten extract solution obtained by step a2 through being evaporated under reduced pressure, recycling design, is obtained into crude extract；
a4：Extraction：By crude extract deionized water dissolving obtained by step a3, extracted 2 times with petroleum ether normal temperature, merge petroleum ether
Part；
a5：Column chromatography for separation is extracted：Step a4 resulting solutions are crossed into polarity silicagel column, use 60% petroleum ether and 40% acetic acid second
Ester mixed liquor elutes, and obtains eluent, is evaporated under reduced pressure and removes solvent；
a6：Resin extracts：By extract obtained 85% ethanol in mass ratio 1 of step a4:1 stirring and dissolving, solution is through D101B types
Macroporous absorbent resin adsorbs 2 times；
a7：Elution：Use concentration for 10%~85% ethanol solution, as low concentration in high concentration to resin described in step a6
Gradient elution, obtain eluent；
a8：Filtering：By eluent obtained by step a7 through the milipore filter that molecular cut off is 1000, filtrate is collected；
a9：Concentration：Ultrafiltrate decompression obtained by step a5 is boiled off into solvent；
a10：Dry：Step a7 gains are placed in vacuum drying chamber, 1.5~3h is dried at 60~80 DEG C.
2. according to claim 1 extract purification shionon technique from beautiful millettia root, it is characterised in that is dried extremely in step a1
Moisture is 8~15%.
3. according to claim 1 extract purification shionon technique from beautiful millettia root, it is characterised in that powder described in step a2
Broken mode is hammer crusher, fluid energy mill, one kind of ball mill.
4. according to claim 1 extract purification shionon technique from beautiful millettia root, it is characterised in that adds described in step a2
Hot leaching mode is one kind in soxhlet extraction method, decocting method.
5. according to claim 1 extract purification shionon technique from beautiful millettia root, it is characterised in that depressurizes and steams in step a3
The vapo(u)rizing temperature evaporated is 65~75 DEG C, and distillation pressure is 50~150Pa.
6. according to claim 1 extract purification shionon technique from beautiful millettia root, it is characterised in that used in step a4
Petroleum ether boiling range be 60~90 DEG C.
7. according to claim 1 extract purification shionon technique from beautiful millettia root, it is characterised in that described in step a6
Large aperture resin is 50~100nm apertures, the D101B type macroporous absorbent resins of 35%~60% cell size.
8. according to claim 1 extract purification shionon technique from beautiful millettia root, it is characterised in that in step a6, resin
It is 25~35 DEG C that adsorption process, which keeps solution temperature, and adsorption flow rate is 2~5BV/h.
9. according to claim 1 extract purification shionon technique from beautiful millettia root, it is characterised in that surpasses described in step a8
Filter membrane material is polyacrylonitrile, polyether sulfone, Kynoar.
10. according to claim 1 extract purification shionon technique from beautiful millettia root, it is characterised in that is filtered in step a8
Pressure is 0.1~0.15MPa.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102219823A (en) * | 2011-06-07 | 2011-10-19 | 南京泽朗农业发展有限公司 | Method for extracting shionone from aster tataricus |
| CN102250193A (en) * | 2011-06-07 | 2011-11-23 | 南京泽朗农业发展有限公司 | Method for preparing shionone by utilizing high-speed countercurrent chromatography |
-
2017
- 2017-06-30 CN CN201710528025.2A patent/CN107383148A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102219823A (en) * | 2011-06-07 | 2011-10-19 | 南京泽朗农业发展有限公司 | Method for extracting shionone from aster tataricus |
| CN102250193A (en) * | 2011-06-07 | 2011-11-23 | 南京泽朗农业发展有限公司 | Method for preparing shionone by utilizing high-speed countercurrent chromatography |
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| Title |
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| 毛秋兰 等: "紫菀中紫菀酮提取工艺的研究", 《临床医学工程》 * |
| 王俊儒 主编: "《天然产物提取分离与鉴定技术》", 31 May 2006, 西北农林科技大学出版社 * |
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Application publication date: 20171124 |