CN103655677A - Novel method for extracting total flavone from humifuse euphorbia herb - Google Patents
Novel method for extracting total flavone from humifuse euphorbia herb Download PDFInfo
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- CN103655677A CN103655677A CN201310706335.0A CN201310706335A CN103655677A CN 103655677 A CN103655677 A CN 103655677A CN 201310706335 A CN201310706335 A CN 201310706335A CN 103655677 A CN103655677 A CN 103655677A
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- total flavones
- herba euphorbiae
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- euphorbiae humifusae
- macroporous adsorbent
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Abstract
The invention belongs to the field of natural organic chemicals and relates to a method for enriching purified total flavone from humifuse euphorbia herb by a supercritical CO2 fluid extraction process and a macroporous resin coupling process. The method is characterized in that the effective ingredients of a medical plant are kept by the supercritical CO2 fluid extraction technology in the extracting process, no chemical reaction occurs in the extraction process, the safety is great, the extracting efficiency is high, less energy is consumed, the production efficiency is increased, and fees and cost are reduced; the purified total flavone is separated by the macroporous absorbing technology, the separating effect is remarkable, the extracting purity is high, a semi-finished total flavone product with more then 30% of total flavone and a finished total flavone product with more than 95% of total flavone can be obtained by the macroporous absorbing technology, and the defects of relatively lower conventional extracting rate and low extracting purity are overcome.
Description
Technical field:
The invention belongs to natural organic chemistry field, relate to a kind of purification process of Herba Euphorbiae Humifusae total flavones, particularly relate to a kind of supercritical CO that utilizes
2the method of total flavones in fluid extraction and macroporous adsorbent resin coupled method enriching and purifying Herba Euphorbiae Humifusae.
Background technology:
The annual draft of crawling of Herba Euphorbiae Humifusae.Stem is very thin, nearly base portion branch, and band aubergine, without hair.Leaf is to life; Petiole is extremely short; Stipule is linear, and common 3 split; Blade Long Circle, long 4-10mm, wide 4-6mm, the blunt circle of tip, base portion is insular, and there is serration at edge, and two sides is without hair or dredge raw pubescence, green or pale red.Cup-shaped inflorescence list is born in axil; Involucre turbination, light red, top 4 is split, sliver Yangtze River Delta shape; Body of gland 4, Long Circle, adularescent petal-shaped appurtenance; Ovary Room 3; Style 3,2 splits.Capsule Rhizoma Sparganii shape is spherical, Glabrous; Seed is avette, and pitchy is about 1.2mm by white wax powder, wide about 0.7mm outward.The florescence 6-10 month, fruit is gradually ripe July.Cure mainly: dysentery; Have loose bowels; Jaundice; Hemoptysis; Spit blood; Hematuria; Have blood in stool; Metrorrhagia; Agalactia; Treating swelling and pain by traumatic injury and pathopyretic ulcer.Ecological environment: being born in Plain, wasteland, roadside and field, is the weeds of commonly seeing.
Herba Euphorbiae Humifusae contains three kinds of flavonoid glycoside, and wherein the glycoside of two glycosides unit is nimbecetin (kaempferol), and the glycoside unit of another glycoside is Quercetin (quercetin).Also containing Coumarins composition: scopoletin (scopoletin), umbelliferone (umbelliferone), A Yapan eupatolide (ayapin).Contain again Palmic acid (palmitic acid), gallic acid (
gallicacid), gallicin (methylgallate) and meso inositol (mesoinositol).In recent years, it is found that in natural plants Herba Euphorbiae Humifusae and contain a large amount of total flavones, because raw material Herba Euphorbiae Humifusae source is wider, therefrom extract total flavones and obtained encouraging progress.At present domestic is mainly that microwave counter current extracts or methanol preparation technology to the extracting method of total flavones, its technique is mainly after dry product is pulverized and uses methanol soaked overnight, using Bu Shi filter cone for filtration, extract 4 times, merging filtrate, last rotary evaporation, concentrating under reduced pressure, spraying are dried to obtain total flavones, and its effect of extraction separation method used is nothing like supercritical CO
2after fluid extraction, carry out the effect of macroporous adsorbent resin separation.
Summary of the invention:
The object of the invention is to overcome the shortcomings such as routine techniques extraction ratio is relatively low, DNA purity is low, a kind of supercritical CO that utilizes is provided
2the method of total flavones in fluid extraction and macroporous adsorbent resin coupled method enriching and purifying Herba Euphorbiae Humifusae.
For achieving the above object, the technical solution used in the present invention is:
A kind of supercritical CO that utilizes
2the method of total flavones in fluid extraction and macroporous adsorbent resin coupled method enriching and purifying Herba Euphorbiae Humifusae, its step is as follows:
(1) dry Herba Euphorbiae Humifusae is pulverized, join CO
2in supercritical extraction device, with ethanol, as entrainer, extracting pressure is 20MPa～40MPa, and temperature is 50 ℃～60 ℃, and extraction time 100min～140min, obtains extract;
(2) the Herba Euphorbiae Humifusae extract of processing through step (1) being adjusted to its pH value with hydrochloric acid is 3.0～4.0, and standing, sucking filtration, concentrating under reduced pressure obtain total flavones crude extract;
(3) gained total flavones crude extract is adsorbed with macroporous adsorbent resin;
(4) with the ethanol of 40%, 50%, 60%, 70%, 80%, 90% variable concentrations, macroporous adsorbent resin is carried out to eluting, by thin layer chromatography, follow the tracks of and detect, collect each stepwise elution liquid of total flavones;
(5) by each stepwise elution liquid concentration and recovery ethanol of total flavones, obtain total flavones semi-finished product, its content is more than 30%;
(6) by silica gel column chromatography on gained total flavones semi-finished product, and with acetone: methanol carries out eluting with 1: 1,2: 1,3: 1,4: 1,5: 1,6: 1,7: 1 different volumes ratios, thin layer chromatography is followed the tracks of and is detected, and collects each stepwise elution liquid of total flavones, concentrating under reduced pressure, reclaims solvent;
(7) by collected total flavones concentrated solution solvent crystallization, fractional crystallization, washing, the dry total flavones of refining to obtain, its content is more than 95%.
In described step (1), Herba Euphorbiae Humifusae was pulverized 80～100 mesh sieves;
In described step (3), macroporous adsorbent resin used is a kind of in D101 type, HPD400 type, S-8, D3520, D4006, H103, D4020, AB-8H and NKA-II type resin, is adsorbed as static adsorption or dynamic adsorption.
The present invention's application supercritical CO
2fluid extraction can guarantee that original color, taste, not because being subject to heat damage, more can guarantee wherein thermal sensitivity, and readily oxidizable substance is not destroyed; Also can in the process of extraction, to extract, carry out separation and purification simultaneously; Macroporous absorption technology separation and purification total flavones, good to the adsorptive selectivity of total flavones, the fast parsing of absorption is also fast, and adsorption capacity is larger; Extract convenient and swiftly, raw material sources are abundant, low production cost, separating effect is obvious, DNA purity is high, can obtain more than 35% total flavones semi-finished product and the more than 95% total flavones finished product of content of content, has overcome the feature that conventional extraction ratio is relatively low, DNA purity is low.
The present invention selects that physicochemical property is stable, surface area is large, exchange velocity is very fast, mechanical strength is high, contamination resistance is strong, the macroporous adsorbent resin of Heat stability is good, be not dissolved in acid, alkali and organic matchmaker, better to Organic substance selectivity, the impact that not existed by inorganic salts and strong ion low molecular compound, can from solution, adsorb selectively by physical absorption total flavones, absorption is fast, parsing is also fast, and adsorption capacity is larger.
The specific embodiment:
Case study on implementation 1: a kind of supercritical CO that utilizes
2the method of total flavones in fluid extraction and macroporous adsorbent resin coupled method enriching and purifying Herba Euphorbiae Humifusae, its step is as follows:
(1) the dry Herba Euphorbiae Humifusae of 1000g is pulverized to rear 80 mesh sieves of crossing, join CO
2in supercritical extraction device, with ethanol, as entrainer, the percent by volume that entrainer accounts for total extractant is 3%, and extracting pressure is 20MPa, and temperature is 55 ℃, CO
2flow is 2mg/g crude drug min, and extraction time 100min, obtains extract;
(2) the Herba Euphorbiae Humifusae extract of processing through step (1) being adjusted to its pH value with hydrochloric acid is 3.0, and standing, sucking filtration, concentrating under reduced pressure obtain total flavones crude extract;
(3) gained total flavones crude extract is adsorbed with macroporous adsorbent resin, macroporous resin used is HPD400 type resin;
(4) with the ethanol of 40%, 50%, 60%, 70%, 80%, 90% variable concentrations, macroporous adsorbent resin is carried out to eluting, by thin layer chromatography, follow the tracks of and detect, collect each stepwise elution liquid of total flavones;
(5) by each stepwise elution liquid concentration and recovery ethanol of total flavones, obtain total flavones semi-finished product, its content is 30.2%;
(6) by silica gel column chromatography on gained total flavones semi-finished product, and with acetone: methanol carries out eluting with 1: 1,2: 1,3: 1,4: 1,5: 1,6: 1,7: 1 different volumes ratios, thin layer chromatography is followed the tracks of and is detected, and collects each stepwise elution liquid of total flavones, concentrating under reduced pressure, reclaims solvent;
(7) by collected total flavones concentrated solution dehydrated alcohol crystallization, fractional crystallization, washing, the dry total flavones 0.169g that refines to obtain, its content is 97.2%.
Case study on implementation 2: a kind of supercritical CO that utilizes
2the method of total flavones in fluid extraction and macroporous adsorbent resin coupled method enriching and purifying Herba Euphorbiae Humifusae, its step is as follows:
(1) the dry Herba Euphorbiae Humifusae of 1000g is pulverized to rear 90 mesh sieves of crossing, join CO
2in supercritical extraction device, with ethanol, as entrainer, the percent by volume that entrainer accounts for total extractant is 4%, and extracting pressure is 30MPa, and temperature is 55 ℃, CO
2flow is 2mg/g crude drug min, and extraction time 120min, obtains extract;
(2) the Herba Euphorbiae Humifusae extract of processing through step (1) being adjusted to its pH value with hydrochloric acid is 3.8, and standing, sucking filtration, concentrating under reduced pressure obtain total flavones crude extract;
(3) gained total flavones crude extract is adsorbed with macroporous adsorbent resin, macroporous resin used is D101 type resin;
(4) with the ethanol of 40%, 50%, 60%, 70%, 80%, 90% variable concentrations, macroporous adsorbent resin is carried out to eluting, by thin layer chromatography, follow the tracks of and detect, collect each stepwise elution liquid of total flavones;
(5) by each stepwise elution liquid concentration and recovery ethanol of total flavones, obtain total flavones semi-finished product, its content is 31.5%;
(6) by silica gel column chromatography on gained total flavones semi-finished product, and with acetone: methanol carries out eluting with 1: 1,2: 1,3: 1,4: 1,5: 1,6: 1,7: 1 different volumes ratios, thin layer chromatography is followed the tracks of and is detected, and collects each stepwise elution liquid of total flavones, concentrating under reduced pressure, reclaims solvent;
(7) by collected total flavones concentrated solution dehydrated alcohol crystallization, fractional crystallization, washing, the dry total flavones 0.197g that refines to obtain, its content is 96.6%.
Case study on implementation 3: a kind of supercritical CO that utilizes
2the method of total flavones in fluid extraction and macroporous adsorbent resin coupled method enriching and purifying Herba Euphorbiae Humifusae, its step is as follows:
(1) the dry Herba Euphorbiae Humifusae of 1000g is pulverized to rear 100 mesh sieves of crossing, join CO
2in supercritical extraction device, with ethanol, as entrainer, the percent by volume that entrainer accounts for total extractant is 6%, and extracting pressure is 40MPa, and temperature is 60 ℃, CO
2flow is 3mg/g crude drug min, and extraction time 140min, obtains extract;
(2) the Herba Euphorbiae Humifusae extract of processing through step (1) being adjusted to its pH value with hydrochloric acid is 4.0, and standing, sucking filtration, concentrating under reduced pressure obtain total flavones crude extract;
(3) gained total flavones crude extract is adsorbed with macroporous adsorbent resin, macroporous resin used is AB-8H type resin;
(4) with the ethanol of 40%, 50%, 60%, 70%, 80%, 90% variable concentrations, macroporous adsorbent resin is carried out to eluting, by thin layer chromatography, follow the tracks of and detect, collect each stepwise elution liquid of total flavones;
(5) by each stepwise elution liquid concentration and recovery ethanol of total flavones, obtain total flavones semi-finished product, its content is 33.7%;
(6) by silica gel column chromatography on gained total flavones semi-finished product, and with acetone: methanol carries out eluting with 1: 1,2: 1,3: 1,4: 1,5: 1,6: 1,7: 1 different volumes ratios, thin layer chromatography is followed the tracks of and is detected, and collects each stepwise elution liquid of total flavones, concentrating under reduced pressure, reclaims solvent;
(7) by collected total flavones concentrated solution dehydrated alcohol crystallization, fractional crystallization, washing, the dry total flavones 0.186g that refines to obtain, its content is 98.3%.
Claims (3)
1. one kind is utilized supercritical CO
2the method of total flavones in fluid extraction and macroporous adsorbent resin coupled method enriching and purifying Herba Euphorbiae Humifusae, its step is as follows:
(1) dry Herba Euphorbiae Humifusae is pulverized, join CO
2in supercritical extraction device, with ethanol, as entrainer, extracting pressure is 20MPa～40MPa, and temperature is 50 ℃～60 ℃, and extraction time 100min～140min, obtains extract;
(2) the Herba Euphorbiae Humifusae extract of processing through step (1) being adjusted to its pH value with hydrochloric acid is 3.0～4.0, and standing, sucking filtration, concentrating under reduced pressure obtain total flavones crude extract;
(3) gained total flavones crude extract is adsorbed with macroporous adsorbent resin;
(4) with the ethanol of 40%, 50%, 60%, 70%, 80%, 90% variable concentrations, macroporous adsorbent resin is carried out to eluting, by thin layer chromatography, follow the tracks of and detect, collect each stepwise elution liquid of total flavones;
(5) by each stepwise elution liquid concentration and recovery ethanol of total flavones, obtain total flavones semi-finished product, its content is more than 30%;
(6) by silica gel column chromatography on gained total flavones semi-finished product, and with acetone: methanol carries out eluting with 1: 1,2: 1,3: 1,4: 1,5: 1,6: 1,7: 1 different volumes ratios, thin layer chromatography is followed the tracks of and is detected, and collects each stepwise elution liquid of total flavones, concentrating under reduced pressure, reclaims solvent;
(7) by collected total flavones concentrated solution solvent crystallization, fractional crystallization, washing, the dry total flavones of refining to obtain, its content is more than 95%.
In described step (1), dry Herba Euphorbiae Humifusae was pulverized 80～100 mesh sieves;
In described step (3), macroporous adsorbent resin used is a kind of in D101 type, HPD400 type, S-8, D3520, D4006, H103, D4020, AB-8H and NKA-II type resin, is adsorbed as static adsorption or dynamic adsorption.
2. the supercritical CO that utilizes according to claim 3
2the method of total flavones in fluid extraction and macroporous adsorbent resin coupled method enriching and purifying Herba Euphorbiae Humifusae, is characterized in that: in described step (1), the percent by volume that entrainer accounts for total extractant is 3%～6%.
3. the supercritical CO that utilizes according to claim 4
2the method of total flavones in fluid extraction and macroporous adsorbent resin coupled method enriching and purifying Herba Euphorbiae Humifusae, is characterized in that: in described step (7), concentrated solution crystallization solvent used is dehydrated alcohol.
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Cited By (2)
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Cited By (3)
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| CN104256604A (en) * | 2014-09-25 | 2015-01-07 | 中国科学院西北高原生物研究所 | Method for extracting and separating total flavonoids from oil-extracted Chinese wolfberry residues |
| CN104257882A (en) * | 2014-09-25 | 2015-01-07 | 中国科学院西北高原生物研究所 | Method for simultaneously extracting and separating oil, chlorogenic acid and total flavonoids from Chinese wolfberry residues |
| CN104257882B (en) * | 2014-09-25 | 2018-02-23 | 中国科学院西北高原生物研究所 | A kind of method for extracting separation grease, chlorogenic acid and general flavone simultaneously from Chinese wolfberry fruit dregs |
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Application publication date: 20140326 |