CN105440095A - Beta-ecdysterone-rich Cyanotis arachnoids extraction and purification method - Google Patents
Beta-ecdysterone-rich Cyanotis arachnoids extraction and purification method Download PDFInfo
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- CN105440095A CN105440095A CN201410432808.7A CN201410432808A CN105440095A CN 105440095 A CN105440095 A CN 105440095A CN 201410432808 A CN201410432808 A CN 201410432808A CN 105440095 A CN105440095 A CN 105440095A
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Abstract
The invention discloses a beta-ecdysterone-rich Cyanotis arachnoids extraction and purification method. The content of beta-ecdysterone in an extract product is 50-100%, and the extract product is prepared from Cyanotis arachnoids through the steps of water refluxing extraction, alcohol precipitation, resin adsorption and recrystallization. The content of the beta-ecdysterone in the Cyanotis arachnoids extract product obtained through adopting the extraction method is greatly improved, and the purity of the extract product reaches 90% or above. The above preparation method is suitable for industrial large-scale production, and is cheap.
Description
Invention field
The invention belongs to effective ingredients in plant extractive technique field, be specifically related to a kind of Radix Cyanotis vagae extracting and purifying method being rich in β-ecdysterone.
Background of invention
Radix Cyanotis vagae (CyanotisarachnoidesC.B.Clarke) belongs to herbaceous plant for many years for the blue ear of Tradescantia albiflora section.Another name has root of Arachnoid Cyanotis, cockscomb ginseng, the wind-weed, the darkish blue Auricled Hedyotis Herb of spider etc., Radix Cyanotis vagae is used as medicine with root, there is dispelling, collateral-activating, inducing diuresis and reducing edema, the effects such as reducing the asthenic fever, be used for the treatment of rheumatic arthritis, numbness of the limbs, pain in the waist and lower extremities, nephritic edema, abnormal heat such as not to move back at the disease, also can external application Zhi Jiao Xian, eczema, knife wound etc.In China, Radix Cyanotis vagae mainly distributes and the areas such as Yunnan, Guangdong, Guangxi, Guizhou, Taiwan, Fujian, and also there are distribution in external India, Sri Lanka, Vietnam, Laos, Cambodia.β-ecdysterone is from Radix Cyanotis vagae, extract obtain main containing a kind of natural steroid compound with insect molting activity, and research finds, Radix Cyanotis vagae be occurring in nature known be rich in the more plant of β-ecdysterone.β-ecdysterone has special physiological function, be the active active substance of natural drug, protective foods, natural cosmetics, the synthesis of human body internal protein especially liver protein can be promoted, the accumulation of body inner cholesterol can be got rid of, suppress blood sugar to rise, and have hypotensive effect; Also can be used for sericulture simultaneously, plant the fields such as the exploitation of source property sterilant.Current China has had Radix Cyanotis vagae preparation to go on the market, but extracts and the restriction of isolation technique due to prior art, and the content of β-ecdysterone is not high, causes drug effect not obvious.The present invention is a kind ofly rich in Cyanotis arachnoides P.E of β-ecdysterone and preparation method thereof fast and efficiently, is very important for making full use of Radix Cyanotis vagae resource, and can also promote Radix Cyanotis vagae plantation.
Summary of the invention
The object of the present invention is to provide the method for the β-ecdysterone of extracting and developing in Radix Cyanotis vagae that a kind of DNA purity is high, yield is high.
Present invention employs following technical scheme:
Be rich in a Radix Cyanotis vagae extracting and purifying method for β-ecdysterone, comprise the following steps:
(1) get without rotting, without going mouldy up-to-standard Radix Cyanotis vagae, rejecting impurity;
(2) get the medicinal material handled well, time little by the water refluxing extraction 1 ~ 5 of volume mass than 5 ~ 40, extraction time is 1 ~ 5 time, filters, merges the filtrate of each time, and is evaporated to raw material weight under 0.09MPa, the condition of 50 ~ 90 DEG C;
(3) concentrated solution is let cool to room temperature, add ethanol and make its determining alcohol reach 50 ~ 80%, let cool, leave standstill, to filter or centrifugal, get alcohol precipitation supernatant liquor and be evaporated between relative density 1.05 ~ 1.25 under 0.09MPa, the condition of 50 ~ 80 DEG C;
(4) alcohol precipitation concentrated solution is crossed macroporous adsorptive resins;
(5) first with water, 5% ethanol carry out wash-out removal of impurities, then use 20 ~ 95% ethanol elutions, retain the elutriant of 20 ~ 95% ethanol.
(6) elutriant of 20 ~ 95% is evaporated to without alcohol taste under 0.09MPa, the condition of 50 ~ 80 DEG C, with the extraction into ethyl acetate of equivalent, retains ethyl acetate layer;
(7) under 0.09MPa, the condition of 50 ~ 80 DEG C, concentrating under reduced pressure obtains ethyl acetate layer, obtains β-ecdysterone crystallization;
(8) be added to by methyl alcohol in β-ecdysterone crystallization, heating makes it dissolve, then in dissolve with methanol liquid, add 100 ~ 300 object column chromatography silica gels of 1 ~ 5 times of sample quality, mixes evaporate to dryness thoroughly, is ground well by the silica gel sample mixed;
(9) by 200 ~ 400 object silica gel column chromatographies on the silica gel sample mixed, with methyl alcohol: ethyl acetate (20:1 ~ 1:20) wash-out, each cut of TLC tracking monitor, collects the cut being rich in β-ecdysterone, concentrating under reduced pressure under 0.09MPa, the condition of 50 ~ 80 DEG C, crystallization;
(10) silicagel column is separated the sample obtained, carries out recrystallization by methyl alcohol, ethyl acetate, place refrigeration, filter, the dry β-ecdysterone sterling obtaining content and be greater than 90%.
Embodiment
Embodiment 1
Select up-to-standard Radix Cyanotis vagae medicinal material, reject impurity, add in extractor, add volume mass than the water being 10, heating and refluxing extraction 1.5 hours, filtrate reduced in volume, adding ethanol makes its alcohol content reach 50%, supernatant liquor concentrating under reduced pressure, HPD100 macroporous resin adsorption post on concentrated solution, removal of impurities is carried out respectively with water, the single volume of each wash-out of 5% ethanol, use the elution of 40% of 6 times amount again, collect elutriant, elutriant concentrates, with the extraction into ethyl acetate 2 times of equivalent, retain ethyl acetate layer; Concentrating under reduced pressure receives dry ethyl acetate, drying, upper silica gel column chromatography column separating purification, cut is collected according to TLC tracking results, concentrating under reduced pressure obtains dry thing, crude product adds volume mass and carries out recrystallization than the ethyl acetate being 8, filters, obtains β-ecdysterone sterling, measure with high performance liquid chromatograph, the content large more than 90% of β-ecdysterone.
Embodiment 2
Select up-to-standard Radix Cyanotis vagae medicinal material, reject impurity, add in extractor, add volume mass than the water being 12, heating and refluxing extraction 1.5 hours, extracts twice, merging filtrate, filtrate reduced in volume, adding ethanol makes its alcohol content reach 60%, supernatant liquor concentrating under reduced pressure, HPD100 macroporous resin adsorption post on concentrated solution, removal of impurities is carried out respectively with water, the single volume of each wash-out of 5% ethanol, again with volume mass than be 6 50% elution, collect elutriant, elutriant concentrate, with the extraction into ethyl acetate 2 times of equivalent, retain ethyl acetate layer; Concentrating under reduced pressure receives dry ethyl acetate, drying, upper silica gel column chromatography column separating purification, cut is collected according to TLC tracking results, concentrating under reduced pressure obtains dry thing, crude product adds volume mass and carries out recrystallization than the ethyl acetate being 8, filters, obtains β-ecdysterone sterling, measure with high performance liquid chromatograph, the content large more than 90% of β-ecdysterone.
Embodiment 3
Select up-to-standard Radix Cyanotis vagae medicinal material, reject impurity, add in extractor, add volume mass than the water being 14, heating and refluxing extraction 1.5 hours, extracts three times, merging filtrate, filtrate reduced in volume, adding ethanol makes its alcohol content reach 70%, supernatant liquor concentrating under reduced pressure, HPD100 macroporous resin adsorption post on concentrated solution, removal of impurities is carried out respectively with water, the single volume of each wash-out of 5% ethanol, again with volume mass than be 6 70% elution, collect elutriant, elutriant concentrate, with the extraction into ethyl acetate 3 times of equivalent, retain ethyl acetate layer; Concentrating under reduced pressure receives dry ethyl acetate, drying, upper silica gel column chromatography column separating purification, cut is collected according to TLC tracking results, concentrating under reduced pressure obtains dry thing, crude product adds volume mass and carries out recrystallization than the ethyl acetate being 10, filters, obtains β-ecdysterone sterling, measure with high performance liquid chromatograph, the content large more than 90% of β-ecdysterone.
Embodiment 4
Select up-to-standard Radix Cyanotis vagae medicinal material, reject impurity, add in extractor, add volume mass than the water being 16, heating and refluxing extraction 2 hours, filtrate reduced in volume, adding ethanol makes its alcohol content reach 80%, supernatant liquor concentrating under reduced pressure, HPD100 macroporous resin adsorption post on concentrated solution, removal of impurities is carried out respectively with water, the single volume of each wash-out of 5% ethanol, again with volume mass than be 6 80% elution, collect elutriant, elutriant concentrate, with the extraction into ethyl acetate 4 times of equivalent, retain ethyl acetate layer; Concentrating under reduced pressure receives dry ethyl acetate, drying, upper silica gel column chromatography column separating purification, cut is collected according to TLC tracking results, concentrating under reduced pressure obtains dry thing, crude product adds volume mass and carries out recrystallization than the ethyl acetate being 12, filters, obtains β-ecdysterone sterling, measure with high performance liquid chromatograph, the content large more than 90% of β-ecdysterone.
Embodiment 5
Select up-to-standard Radix Cyanotis vagae medicinal material, reject impurity, add in extractor, add volume mass than the water being 16, heating and refluxing extraction 2 hours, extracts twice, merging filtrate, filtrate reduced in volume, adding ethanol makes its alcohol content reach 70%, supernatant liquor concentrating under reduced pressure, HPD600 macroporous resin adsorption post on concentrated solution, removal of impurities is carried out respectively with water, the single volume of each wash-out of 5% ethanol, again with volume mass than be 6 40% elution, collect elutriant, elutriant concentrate, with the extraction into ethyl acetate 4 times of equivalent, retain ethyl acetate layer; Concentrating under reduced pressure receives dry ethyl acetate, drying, upper silica gel column chromatography column separating purification, cut is collected according to TLC tracking results, concentrating under reduced pressure obtains dry thing, crude product adds volume mass and carries out recrystallization than the ethyl acetate being 10, filters, obtains β-ecdysterone sterling, measure with high performance liquid chromatograph, the content large more than 90% of β-ecdysterone.
Claims (4)
1. one kind is rich in the Radix Cyanotis vagae extracting and purifying method of β-ecdysterone, it is characterized in that: containing β-ecdysterone 50% ~ 100%, take Radix Cyanotis vagae as raw material, by comprise water refluxing extraction, alcohol precipitation, resin absorption, silica gel column chromatography, purification procedures obtain, specifically comprise:
A, Feedstock treating: select life in 1 ~ 2 year, without rotting, without to go mouldy up-to-standard Radix Cyanotis vagae, rejecting impurity;
B, water refluxing extraction: the raw material after process is put into extractor, adds the water of volume mass than 5 ~ 40, heating and refluxing extraction 1 ~ 5 hour, extract 1 ~ 5 time, filter, merging filtrate;
C, the filtrate after extracting is evaporated to the concentrated solution of 0.5 ~ 3g raw medicinal herbs/ml at 50 ~ 90 DEG C, adding ethanol makes its alcohol content reach 50 ~ 90%, leaves standstill, filtration or centrifugal, get supernatant liquor, alcohol precipitation supernatant liquor is evaporated to the concentrated solution of 0.5 ~ 3g raw medicinal herbs/ml at 50 ~ 80 DEG C;
D, resin absorption: macroporous resin adsorption post on alcohol precipitation concentrated solution, control column flow rate 80 ~ 100kg/h, wash 1 ~ 10 times of column volume with water, use the ethanol elution removal of impurities of 1 ~ 10 times of column volume 5% ~ 20% again, finally use the ethanol elution of 20 ~ 95% of 1 ~ 12 times of column volume, collect elutriant, reclaim ethanol;
E, the elutriant of 30 ~ 90% is evaporated to without alcohol taste under 0.09MPa, the condition of 50 ~ 90 DEG C, with extraction into ethyl acetate 1-6 time of equivalent, retains ethyl acetate layer, concentrated crystallization;
F, silica gel column chromatography: dissolve adding solvothermal in step e coarse-grain, add the column chromatography silica gel of 0.5 ~ 5 times of example weight, evaporate to dryness, porphyrize, be laid in column chromatography silica gel post upper strata, with methyl alcohol: ethyl acetate (20:1 ~ 1:20) mixed solution carries out wash-out, TLC follows the tracks of, and collect elutriant, evaporate to dryness analyses coarse-grain;
G, purifying: carry out recrystallization by adding the ethyl acetate of volume mass than 6 ~ 12 in step F crude product, filter, obtain β-ecdysterone sterling through cryodrying.
2. a kind of Radix Cyanotis vagae extracting and purifying method being rich in β-ecdysterone according to claim, is characterized in that: the amount of water volume mass ratio of described step B is 5 ~ 40; Extraction time is 1 ~ 5h, and extraction time is 1 ~ 5 time.
3. a kind of Radix Cyanotis vagae extracting and purifying method being rich in β-ecdysterone according to claim, it is characterized in that: the macroporous resin type polymeric adsorbent of described D step includes but not limited to: D101, AB-8, HPD100, HPD400, HPD100, HPD600, D941 type macroporous resin.
4. a kind of Radix Cyanotis vagae extracting and purifying method being rich in β-ecdysterone according to claim, is characterized in that: the solvent of described F step is methyl alcohol, ethanol, anhydrous methanol, dehydrated alcohol, N.N-dimethyl formamide etc.
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Cited By (4)
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| CN107727777A (en) * | 2017-12-04 | 2018-02-23 | 山东师范大学 | A kind of method and HPTLC scanning method detection method that β moulting hormones are extracted from krill |
| CN112851737A (en) * | 2021-01-12 | 2021-05-28 | 四川兴杰象药业有限公司 | System and method for efficiently extracting ecdysone |
| CN113943335A (en) * | 2021-11-25 | 2022-01-18 | 云南西草生物科技开发有限公司 | Preparation process of herba Cymbopogonis Citrari extract |
| CN113956319A (en) * | 2021-09-23 | 2022-01-21 | 西安尚诚生物科技有限公司 | Method for extracting and refining ecdysterone |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107727777A (en) * | 2017-12-04 | 2018-02-23 | 山东师范大学 | A kind of method and HPTLC scanning method detection method that β moulting hormones are extracted from krill |
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| CN112851737A (en) * | 2021-01-12 | 2021-05-28 | 四川兴杰象药业有限公司 | System and method for efficiently extracting ecdysone |
| CN113956319A (en) * | 2021-09-23 | 2022-01-21 | 西安尚诚生物科技有限公司 | Method for extracting and refining ecdysterone |
| CN113956319B (en) * | 2021-09-23 | 2024-04-09 | 西安尚诚生物科技有限公司 | Method for extracting and refining ecdysterone |
| CN113943335A (en) * | 2021-11-25 | 2022-01-18 | 云南西草生物科技开发有限公司 | Preparation process of herba Cymbopogonis Citrari extract |
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