CN103159820A - Method using microwave reverse flow technique to extract marsh marigold lactone in marsh marigold - Google Patents
Method using microwave reverse flow technique to extract marsh marigold lactone in marsh marigold Download PDFInfo
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- CN103159820A CN103159820A CN2012105955857A CN201210595585A CN103159820A CN 103159820 A CN103159820 A CN 103159820A CN 2012105955857 A CN2012105955857 A CN 2012105955857A CN 201210595585 A CN201210595585 A CN 201210595585A CN 103159820 A CN103159820 A CN 103159820A
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Abstract
The invention belongs to the field of natural organic chemistry. Microwave reverse flow is adopted to carry out extraction. The method has the advantages that heating is even, a process is easy to control, preprocessing such as drying is not needed to be carried out, a technology is simplified, investment is reduced, good extraction efficiency is greatly improved, macroporous resin adsorption-column chromatographic technique is adopted to separate and purify the marsh marigold lactone, adsorptive selectivity to the marsh marigold lactone is good, absorption is rapid, desorption is rapid, adsorption capacity is quite large, extraction is convenient and rapid, production cost is low, separation effect is obvious, purity of extraction is high, the marsh marigold lactone can be selectively absorbed from the solution through physical absorption, more than 20% of semi-finished products of the marsh marigold lactone and end products of the marsh marigold lactone are obtained, wherein the end products of the marsh marigold lactone are more than 70% in content, and the defects that conventional extraction ratio is relevantly low and purity of extraction is low are overcome.
Description
Technical field:
The invention belongs to the natural organic chemistry field, relate to a kind of purification process of kincup lactone, particularly relate to a kind of method of utilizing kincup lactone in microwave counter current extraction-macroporous resin adsorption-column chromatography coupling technology enriching and purifying kincup.
Background technology:
Kincup: English name Herb of Membranaceous Marshmarigold, another name: donkey hoof dish, marsh marigold.Perennial herb, high 20-48cm is without hair, fibrous root meat.Stem is upright, and is solid, the thin longitudinal furrow of tool, and middle part or middle part be with upper branch, rare not branch.Basal leaf 3-7, herbaceous stem has long handle; The long 7-24cm of handle; Blade is circular, circle kidney shape or with shape, long 2.5-5cm, and wide 3-9cm, tip is justified, base portion innermost being shape, the close living denticle in edge; Stem leaf is less, tool short handle or stockless.Single over-expense cyme is born in stem or branch top, and 2 flowers are arranged usually; The long 2-10cm of bennet; The flower both sexes, diameter 1.6-3.2cm; Sepal 5, petal-shaped, yellow, obovate or narrow obovate, long 1-1.8cm, wide 0.6-1.2cm, tip circle; Petal without; Stamen is most, long 4.5-7mm, and filigree is narrow linear, flower pesticide Long Circle, long 1-1.6mm; Carpel 7-12, closely isometric with stamen, stockless, style Duan ， Follicle radish fruit is about 1cm, and wide approximately 3mm has the transverse vein line, and beak is about 1mm.Seed is most, narrow ovoid, and long 1.5-2mm, black is glossy, and the tool minority is indulged wrinkle.The florescence 5-9 month, the fruit phase 6-10 month.Chemical composition: herb contains corytuberine (corytuberine), magnoflorine (magnoflorine), nicotine (nicotine), kincup lactone (caltholide), the compositions such as table kincup lactone (epiacltholide).
The kincup lactone is a kind of natural active matter that human health is had remarkable health-care effect, has caused the common concern of international community.Prove through scientific research and clinical application, the kincup lactone has clearing heat and promoting diuresis, detoxifcation.Be used for heatstroke, urinary tract infections; Burn and scald is controlled in external application, venomous snake bite.In recent years, it is found that and contain a large amount of kincup lactones in the natural phant kincup, therefrom extract the kincup lactone and obtained encouraging progress, raw material kincup source is wider.
Summary of the invention:
The object of the invention is to overcome the shortcomings such as the routine techniques extraction yield is relatively low, DNA purity is low, a kind of method of utilizing microwave counter current to extract kincup lactone in macroporous resin adsorption-column chromatography coupling technology enriching and purifying kincup is provided.
For achieving the above object, the technical solution used in the present invention is:
A kind of method of utilizing kincup lactone in microwave counter current extraction-macroporous resin adsorption-column chromatography coupling technique enriching and purifying kincup, its step is as follows:
(1) with after dried kincup pulverizing, add the pure water mixing, carry out microwave counter current and extract, forward feeding, oppositely water inlet, the mobile of material and water is reverse continuous dynamic flow, and microwave power is at 400～800W, and extraction time is 60min～120min; Extraction temperature is 50～60 ℃, extracts three times, filters, and merging filtrate obtains extracting solution;
(2) will through the kincup extracting solution suction filtration of step (1) processing, get extracting solution and concentrating under reduced pressure and get the kincup primary extract:
(3) with obtaining primary extract water heating for dissolving, add ethyl acetate extraction, standing, suction filtration, concentrating under reduced pressure get kincup lactone crude extract;
(4) gained kincup lactone crude extract is adsorbed with macroporous adsorbent resin;
(5) ethanol with different concns carries out wash-out to macroporous adsorbent resin, follows the tracks of with tlc and detects, and collects each stepwise elution liquid of kincup lactone;
(6) with each stepwise elution liquid concentration and recovery ethanol of kincup lactone, get kincup lactone work in-process, its content is more than 20%;
(7) gained kincup lactone work in-process are dissolved in ethyl acetate and add the proper silica gel stirring, dried the 60-80 mesh sieve, the dry method loading, upper technical grade post 200-300 order silica gel column chromatography, and carry out wash-out with ethanol, ether with the different volumes ratio, tlc is followed the tracks of and is detected, and collects each stepwise elution liquid of kincup lactone;
(8) with collected kincup lactone elutriant, adopt activated carbon decolorizing, recrystallization method to make with extra care to get the kincup lactone, its content is more than 70%.
In described step (1), after pulverizing, kincup crosses 60～80 mesh sieves.
In described step (1), the volume that adds purified water is 8～10 times that kincup is pulverized volume.
In described step (3), add the amount of ethyl acetate by volume to calculate to be 10～20 times after the extract dissolving.
In described step (4), macroporous adsorbent resin used is a kind of in Tianjin Nankai university X-5, the NKA-9, S-8, D3520, D4006, H103, D4020, AB-8H and the NKA-II type resin that produce, is adsorbed as Static Adsorption or dynamic adsorption.
Wash-out described in described step (5) carries out wash-out for the ethanol with 10～90%.
In described step (5) and (7), the detection method of taking is tlc and follows the tracks of detection.
The ethanol of taking in described step (7) and the volume of ether are 1～5: 1.
The present invention adopts microwave counter current to extract, and has homogeneous heating, process is easy to control, and need not the dry pre-treatment that waits, and has simplified technique, has reduced investment, has greatly improved extraction efficiency; Macroporous adsorption resin technology and column chromatographic isolation and purification kincup lactone, adsorption selectivity to the kincup lactone is good, the fast parsing of absorption is also fast, and loading capacity is larger, extracts convenient and swift, raw material sources are abundant, low production cost, separating effect is obvious, and DNA purity is high, can obtain kincup lactone work in-process and the content kincup lactone finished product 70% or more of content more than 20%, overcome the shortcoming that conventional extraction yield is relatively low, DNA purity is low.
The present invention selects that physico-chemical property is stable, surface-area is large, exchange velocity is very fast, physical strength is high, contamination resistance is strong, the macroporous adsorbent resin of Heat stability is good, be insoluble to acid, alkali and organic matchmaker, better to the organism selectivity, the impact that not existed by inorganic salts and strong ion low molecular compound, can adsorb selectively by physical adsorption the kincup lactone from solution, absorption is fast, resolve also fast, loading capacity is larger.
Embodiment:
Embodiment 1: a kind of method of utilizing kincup lactone in microwave counter current extraction-macroporous adsorbent resin-column chromatography coupling technology enriching and purifying kincup, and its step is as follows:
(1) will cross 60 mesh sieves after the dried kincup pulverizing of 500g, the pure water that adds kincup to pulverize 10 times of volumes carries out microwave counter current and extracts, forward feeding, reverse water inlet, and flowing of material and water is reverse continuous dynamic flow.Microwave power is at 400W, and extraction time is 60min; Extraction temperature is 50 ℃, extracts three times, filters, and merging filtrate obtains extracting solution;
(2) will through the kincup extracting solution suction filtration of step (1) processing, get extracting solution and concentrating under reduced pressure and get the kincup primary extract;
(3) with obtaining primary extract water heating for dissolving, adding volume is that the ethyl acetate of 10 times of solution of extract volumes extracts, and standing, suction filtration, concentrating under reduced pressure get kincup lactone crude extract;
(4) gained kincup lactone extract is adsorbed with macroporous adsorbent resin, macroporous resin used is the NKA-II type resin that Tianjin Nankai university produces;
(5) ethanol with 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% different concns carries out gradient elution successively in macroporous adsorbent resin layer post, follow the tracks of with tlc and detect, collect each stepwise elution liquid of kincup lactone;
(6) with each stepwise elution liquid concentration and recovery ethanol of kincup lactone, obtain kincup lactone work in-process, its content is 23.1%;
(7) gained kincup lactone work in-process are dissolved in ethyl acetate and add the proper silica gel stirring, dried 60 mesh sieves, the dry method loading, upper technical grade post 200-300 order silica gel column chromatography, and carry out gradient elution with the volume ratio of 1: 1,2: 1,3: 1,4: 1,5: 1 successively with ethanol, ether, tlc is followed the tracks of and is detected, and collects each stepwise elution liquid of kincup lactone;
(8) with collected kincup lactone elutriant, adopt activated carbon decolorizing, recrystallization method to make with extra care and obtain the kincup lactone, its content is 72.3%.
Case study on implementation 2: a kind of method of utilizing kincup lactone in microwave counter current extraction-macroporous adsorbent resin-column chromatography coupling technology enriching and purifying kincup, its step is as follows:
(1) will cross 70 mesh sieves after the dried kincup pulverizing of 500g, the pure water that adds kincup to pulverize 15 times of volumes carries out microwave counter current and extracts, forward feeding, reverse water inlet, and flowing of material and water is reverse continuous dynamic flow.Microwave power is at 600W, and extraction time is 90min; Extraction temperature is 55 ℃, extracts three times, filters, and merging filtrate obtains extracting solution;
(2) will through the kincup extracting solution suction filtration of step (1) processing, get extracting solution and concentrating under reduced pressure and get the kincup primary extract;
(3) with obtaining primary extract water heating for dissolving, adding volume is that the ethyl acetate of 15 times of solution of extract volumes extracts, and standing, suction filtration, concentrating under reduced pressure get kincup lactone crude extract;
(4) gained kincup lactone extract is adsorbed with macroporous adsorbent resin, macroporous resin used is the AB-8H type resin that Tianjin Nankai university produces;
(5) ethanol with 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% different concns carries out gradient elution successively in macroporous adsorbent resin layer post, follow the tracks of with tlc and detect, collect each stepwise elution liquid of kincup lactone;
(6) with each stepwise elution liquid concentration and recovery ethanol of kincup lactone, obtain kincup lactone work in-process, its content is 23.7%:
(7) gained kincup lactone work in-process are dissolved in ethyl acetate and add the proper silica gel stirring, dried 80 mesh sieves, the dry method loading, upper technical grade post 200-300 order silica gel column chromatography, and carry out gradient elution with the volume ratio of 1: 1,2: 1,3: 1,4: 1,5: 1 successively with ethanol, ether, tlc is followed the tracks of and is detected, and collects each stepwise elution liquid of kincup lactone;
(8) with collected kincup lactone elutriant, adopt activated carbon decolorizing, recrystallization method to make with extra care and obtain the kincup lactone, its content is 74.5%.
Case study on implementation 3: a kind of method of utilizing kincup lactone in microwave counter current extraction-macroporous adsorbent resin-column chromatography coupling technology enriching and purifying kincup, its step is as follows:
(1) will cross 80 mesh sieves after the dried kincup pulverizing of 500g, the pure water that adds kincup to pulverize 20 times of volumes carries out microwave counter current and extracts, forward feeding, reverse water inlet, and flowing of material and water is reverse continuous dynamic flow.Microwave power is at 800W, and extraction time is that the 120min extraction temperature is 60 ℃, extracts three times, filters, and merging filtrate obtains extracting solution;
(2) will through the kincup extracting solution suction filtration of step (1) processing, get extracting solution and concentrating under reduced pressure and get the kincup primary extract;
(3) with obtaining primary extract water heating for dissolving, adding volume is that the ethyl acetate of 20 times of solution of extract volumes extracts, and standing, suction filtration, concentrating under reduced pressure get kincup lactone crude extract;
(4) gained kincup lactone extract is adsorbed with macroporous adsorbent resin, macroporous resin used is the NKA-9 type resin that Tianjin Nankai university produces;
(5) ethanol with 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% different concns carries out gradient elution successively in macroporous adsorbent resin layer post, follow the tracks of with tlc and detect, collect each stepwise elution liquid of kincup lactone;
(6) with each stepwise elution liquid concentration and recovery ethanol of kincup lactone, obtain kincup lactone work in-process, its content is 24.2%:
(7) gained kincup lactone work in-process are dissolved in ethyl acetate and add the proper silica gel stirring, dried 80 mesh sieves, the dry method loading, upper technical grade post 200-300 order silica gel column chromatography, and carry out gradient elution with the volume ratio of 1: 1,2: 1,3: 1,4: 1,5: 1 successively with ethanol, ether, tlc is followed the tracks of and is detected, and collects each stepwise elution liquid of kincup lactone;
(8) with collected kincup lactone elutriant, adopt activated carbon decolorizing, recrystallization method to make with extra care and obtain the kincup lactone, its content is 75.3%.
Claims (6)
1. method of utilizing kincup lactone in microwave counter current extraction-macroporous resin adsorption-column chromatography coupling technique enriching and purifying kincup, its step is as follows:
(1) with after dried kincup pulverizing, add the pure water mixing, carry out microwave counter current and extract, forward feeding, oppositely water inlet, the mobile of material and water is reverse continuous dynamic flow.Microwave power is at 400～800W, and extraction time is 60min～120min; Extraction temperature is 50～60 ℃, extracts three times, filters, and merging filtrate obtains extracting solution;
(2) will through the kincup extracting solution suction filtration of step (1) processing, get extracting solution and concentrating under reduced pressure and get the kincup primary extract;
(3) with obtaining primary extract water heating for dissolving, add ethyl acetate extraction, standing, suction filtration, concentrating under reduced pressure get kincup lactone crude extract;
(4) gained kincup lactone crude extract is adsorbed with macroporous adsorbent resin;
(5) ethanol with different concns carries out wash-out to macroporous adsorbent resin, follows the tracks of with tlc and detects, and collects each stepwise elution liquid of kincup lactone;
(6) with each stepwise elution liquid concentration and recovery ethanol of kincup lactone, get kincup lactone work in-process, its content is more than 20%;
(7) gained kincup lactone work in-process are dissolved in ethyl acetate and add the proper silica gel stirring, dried the 60-80 mesh sieve, the dry method loading, upper technical grade post 200-300 order silica gel column chromatography, and carry out wash-out with ethanol, ether with the different volumes ratio, tlc is followed the tracks of and is detected, and collects each stepwise elution liquid of kincup lactone;
(8) with collected kincup lactone elutriant, adopt activated carbon decolorizing, recrystallization method to make with extra care to get the kincup lactone, its content is more than 70%.
In described step (4), macroporous adsorbent resin used is a kind of in Tianjin Nankai university X-5, the NKA-9, S-8, D3520, D4006, H103, D4020, AB-8H and the NKA-II type resin that produce, is adsorbed as Static Adsorption or dynamic adsorption.
In described step (5) and (7), the detection method of taking is tlc and follows the tracks of detection.
2. the method for utilizing kincup lactone in microwave counter current extraction-macroporous resin adsorption-column chromatography coupling technique enriching and purifying kincup according to claim 1, its speciality is: in described step (1), cross 60～80 mesh sieves after kincup is pulverized.
3. the microwave counter current extraction-macroporous resin adsorption of utilizing according to claim 2--the method for kincup lactone in column chromatography coupling technique enriching and purifying kincup, it is characterized in that: in described step (1), the volume that adds purified water is 10～20 times that kincup is pulverized volume.
4. the method for utilizing kincup lactone in microwave counter current extraction-macroporous resin adsorption-column chromatography coupling technique enriching and purifying kincup according to claim 3, is characterized in that; In described step (3), add the amount of ethyl acetate to be 10～20 times after the extract dissolving by volume.
5. the method for utilizing kincup lactone in microwave counter current extraction-macroporous resin adsorption-column chromatography coupling technique enriching and purifying kincup according to claim 4, it is characterized in that: in described step (5), described elutriant carries out wash-out with 10～90% aqueous ethanolic solution.
6. the method for utilizing kincup lactone in microwave counter current extraction-macroporous resin adsorption-column chromatography coupling technique enriching and purifying kincup according to claim 5, it is characterized in that: in described step (7), the volume ratio of the ethanol that adopts, ether is 1～5: 1.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101062069A (en) * | 2007-06-18 | 2007-10-31 | 石任兵 | Whole coumarins extract from root of dahuriae angelica and the preparing method thereof |
| CN101073590A (en) * | 2007-06-18 | 2007-11-21 | 石任兵 | Muxiang total lactone extract and its production |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101062069A (en) * | 2007-06-18 | 2007-10-31 | 石任兵 | Whole coumarins extract from root of dahuriae angelica and the preparing method thereof |
| CN101073590A (en) * | 2007-06-18 | 2007-11-21 | 石任兵 | Muxiang total lactone extract and its production |
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