CN101209284A - Method for continuously extracting alkaloids, flavone and polysaccharide active components from mulberry leaves - Google Patents
Method for continuously extracting alkaloids, flavone and polysaccharide active components from mulberry leaves Download PDFInfo
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Abstract
The invention relates to a method of taking mulberry leaves as the raw materials to continuously extract the active ingredients of alkaloids, flavonoids and polysaccharides. The invention includes the following steps: the solvents of 20 percent to 90 percent ethanol or 10 percent to 90 percent acetone are selected for reflux extraction to obtain the filtrate and filter residue; the filtrate is concentrated and dissolved by adding water, the flavonoid compounds with the content of 30 percent to 50 percent are obtained by macroporous resin separation; the solution is treated by ion exchange resin separation to get the alkaloid compounds with the content of 40 percent to 60 percent after passes through the macroporous resin; the filter residue of the mulberry leaves is boiled and decolored by using the water, and the acid polysaccharide compounds with the content of 55 percent to 85 percent are obtained by 10 percent to 60 percent alcohol sedimentation. The invention continuously and efficiently separates three active ingredients of alkaloids, flavonoids and polysaccharides of the mulberry leaves by one-time feeding, all the steps are not disturbed mutually, the extraction rate is high, the process operation is simple, the cost is low, and the invention is conductive to industrial popularization and application.
Description
Technical field
The present invention relates to a kind of from Folium Mori the method for continuously extracting alkaloids, flavone and polysaccharide active components, belong to the technology of from natural product, extracting bioactive ingredients.
Background technology
Folium Mori different name Herba adianti myriosori is the leaf of moraceae plants Mulberry (Morus alba.L), and bitter in the mouth is sweet, cold, is distributed in most of area, the whole nation.Modern pharmacological research proof Folium Mori can suppress blood sugar increasing, can prevent and treat diabetes.In addition, Folium Mori also have the effect of expelling wind and clearing away heat, cooling blood for improving eyesight, and the property of medicine is gentle and have no side effect, and formally are included into the row of " be food be again medicine " by health ministry.
Contain the various active composition in the Folium Mori, comprise alkaloid, polysaccharide, flavone, polyphenol, aminoacid and various trace elements, wherein topmost active component is alkaloid, polysaccharide and flavone.Asano N isolation identification from Folium Mori goes out 11 kinds of alkaloids, and wherein the content of 1-deoxynojirimycin (DNJ) in Folium Mori reaches 0.1%, far above other plant, is one of effective inhibitors of alpha-glucosidase, has very strong hypoglycemic activity.The Folium Mori total polysaccharides also is the main active in the Folium Mori, and the alloxan diabetes mice is had significant hypoglycemic activity.General flavone content accounts for 1.0%～3.0% of Folium Mori dry weight in the Folium Mori, mainly contains rutin, isoquercitrin, and Quercetin-3-tri-glucose glycosides and Herba Astragali Melilotoidis (Herba Astragali Sinici) glycosides etc. have antioxidation, suppress effects such as serum lipids increase and the formation of inhibition atherosclerosis.There are some researches show that flavones in mulberry leaves also has hypoglycemic activity, and flavone is combined with DNJ, more can suppress the rising of blood glucose than independent DNJ effectively.And therefore alkaloid, polysaccharide and the flavonoid active effective part in extraction separation and the purification Folium Mori made Chinese patent medicine or functional health care food all has broad prospects.
About the more existing relevant patents in the separation aspect of active effective part in the Folium Mori, as the domestic patent No. 03101988.9 " a kind of preparation that contains Folium Mori total alkali extractum and preparation method thereof ", the alkaloidal preparation method in a kind of Folium Mori is disclosed at present; The patent No. 200510012388.8 " a kind of flavones in mulberry leaves extracting method " discloses a kind of method of separating the flavone in the Folium Mori; The patent No. 01129976.2 " a kind of mulberry leaf polyose and uses thereof " discloses a kind of extraction process of polysaccharide; The patent No. 200610038713.2 " composition of white mulberry leaf blood-sugar-reducing effective components and preparation method " discloses a kind of alkaloid in the Folium Mori and method of polysaccharide of separating.But these patents only can be isolated one or both effective ingredient, and other effective ingredient is then had to be used as impurity and removed in purge process, not only caused the very big wasting of resources, and product cost is also higher.And existing patent all is the alkaloid in the solvent extraction Folium Mori with water, and our research experiment shows, water is lower than the extraction ratio of finite concentration solvent to the alkaloid extraction rate, and complicated component in the water extract, has brought difficulty for follow-up separation.In addition, at present general to the polysaccharides of Folium Mori general concept that extracts, the analysis of its structure and classification be yet there are no report.
Summary of the invention
The purpose of this invention is to provide a kind of from Folium Mori the method for continuously extracting alkaloids, flavone and polysaccharide active components, this procedure is simple, alkaloid, flavone and polyoses content height in the extract.
The present invention realized by the following technical programs, a kind of from Folium Mori the method for continuously extracting alkaloids, flavone and polysaccharide active components, it is characterized in that comprising following process:
1) volume ratio by dry Folium Mori weight and solution is 1: 10～15, it is 20%～90% alcoholic solution that dry Folium Mori after pulverizing are soaked in volumetric concentration, or to be soaked in volumetric concentration be 10%～90% acetone soln, reflux, extract, is 3 times under 95 ℃ of conditions, each 30min that extracts, extracting solution is mixed and filtration under diminished pressure, obtain filtrate and filtering residue.
2) step 1) gained filtrate is removed ethanol through reduction vaporization, the volume ratio by dry Folium Mori weight and deionized water is 1: 10～15 adding deionized waters then, leaves standstill 2h, centrifugalize.Supernatant is by macroporous resin X-5, D101, D4020 or H103,20～30 ℃ of absorption, at first use ionized water eluting macroporous resin, then with ethanol or acetone soln eluting macroporous resin, collect eluent, eluent is through evaporation under reduced pressure removed ethanol or acetone, obtains mass content and be 30%～50% flavone compound, and the main component of flavone is rutin and isoquercitrin.
3) stream is through the solution behind the macroporous resin, by model is 001 * 7,001 * 4 or 001 * 7 * 7 cation exchange resin is at 20～30 ℃ of absorption 5～10h, use earlier the deionized water rinsing resin, obtain eluent with ammonia eluting resin again, it is residual that eluent is concentrated into no ammonia, be 201 * 7 by model again, D201 or the absorption of 201 * 4 anion exchange resin, get eluent with deionized water eluting anion exchange resin, eluent is through reduction vaporization, obtain mass content and be 40%～60% alkaloid compound, alkaloidal main component is 1-deoxynojirimycin (DNJ).
4) the Folium Mori filtering residue that obtains in the step 1) deionized water with 10～15 times of volumes is decocted twice down at 60～90 ℃, filtration under diminished pressure, collect filtrate, filtrate is through activated carbon decolorizing, filtrate decompression after the decolouring is concentrated into 1/5～1/10 of extracting liquid volume, adds ethanol afterwards in concentrated solution, makes that the ethanol volumetric concentration is 10%～60% in the concentrated solution, leave standstill 10～15h, centrifugalize gets precipitate.Use dehydrated alcohol, acetone and ether washing precipitate more successively, obtain mass content and be 55%～85% polysaccharide compound, this compound of polysaccharide main component is an acidic polysaccharose, its molecular weight 500,000～550,000Da.
The invention has the advantages that, 20%～90% ethanol of selecting for use or 10%～90% acetone soln have good leaching ability to alkaloid in the Folium Mori and flavone, and to polysaccharide leaching ability in the Folium Mori, in addition, the macroporous resin that is adopted in the process has stronger absorbability to flavone, but relatively poor to the alkaloid absorbability, so can be continuously with the alkaloid in the Folium Mori, flavone and separation of polysaccharides in the process, and the rate of departure is fast, component content height in the product.
Description of drawings
The liquid chromatogram of the flavone that Fig. 1 is extracted for the embodiment of the invention 2, peak 1 is the rutin composition among the figure, peak 2 is the isoquercitrin composition.
The alkaloidal liquid chromatogram that Fig. 2 is extracted for the embodiment of the invention 2, peak 1 is a 1-deoxynojirimycin composition among the figure.
Fig. 3 by the embodiment of the invention 2 the HP-GPC figure of extraction polysaccharide, peak 1 is a polysaccharide component among the figure.
The specific embodiment
Example 1
Folium Mori raw material 400g after pulverizing is added 30% ethanol water of 4L, and reflux, extract, under 95 ℃ of conditions repeats to extract 3 times, each 30min, and the extracting solution that obtains for three times is respectively 2.9L, 3.7L, 4.1L, and merge extractive liquid, filters.Filtering residue dries standby, and ethanol evaporate to dryness in the gained filtrate is got extractum, adds the 2L deionized water then in extractum it is dissolved, and leaves standstill 2h, centrifugalize.Supernatant adsorbs 5h by 200g D4020 macroporous resin under 25 ℃ of conditions.Then earlier with 2L deionized water rinsing macroporous resin, 60% the ethanol water eluting macroporous resin of reuse 1L is collected ethanol elution, and the evaporated under reduced pressure eluent obtains extractum 6.3g, and wherein the mass content of total flavones is 31%.
Flow through solution behind the macroporous resin, 001 * 4 cation exchange resin by 50g adsorbs 5h down at 25 ℃, earlier with 500ml deionized water rinsing resin, get eluent with 150ml ammonia eluting resin again, eluent is concentrated into 30ml, get eluent after concentrating again by 201 * 4 anion exchange resin (50g), and with 500ml deionized water eluting anion exchange resin, eluent gets extractum 1.3g through reduction vaporization, and wherein the mass content of alkaloid DNJ is 43.0%.
With the Folium Mori filtering residue behind intact alkaloid of said extracted and the flavone is raw material, adds the 3L deionized water, puts into 70 ℃ of waters bath with thermostatic control, lixiviate 1.5h, extract 2 times, extracted twice liquid is respectively 1.8L and 2.7L, and merge extractive liquid, also filters, regulate filtrate pH3.0, filtrate is concentrated into 500ml behind the 200g activated carbon decolorizing, add ethanol in the extracting solution after concentrating then, and regulating the ethanol volumetric concentration is 40%, leave standstill 12h, centrifugalize must precipitate.Adopt dehydrated alcohol, acetone and ether washing precipitate successively, get Folium Mori total polysaccharides precipitation 6.2g, wherein neutral polysaccharide content 21%, and acidic polysaccharose content is 51.3%.
Example 2
70% ethanol water that Folium Mori raw material 400g after pulverizing is added 4L, 95 ℃ of reflux, extract, 3 times, each 30min, the extracting solution that obtains three times is respectively 2.8L, 3.3L, 3.8L, and merge extractive liquid, also filters, and filtering residue dries standby, ethanol evaporate to dryness in the gained filtrate is got extractum, in extractum, add the 4L distilled water then, leave standstill 2h, centrifugalize.Supernatant adsorbs under 30 ℃ of conditions by the H-103 macroporous resin of 200g.Then earlier with 2L deionized water rinsing macroporous resin, 60% the aqueous acetone solution eluting macroporous resin of reuse 1L, the evaporated under reduced pressure eluent obtains extractum 7.6g, and wherein the mass content of total flavones is 36%.
Flow through solution behind the macroporous resin, 001 * 7 cation exchange resin by 60g adsorbs 5h down at 25 ℃, earlier with 600ml deionized water rinsing resin, get eluent with 300ml ammonia eluting resin again, eluent is concentrated into 50ml, get eluent after concentrating again by 201 * 7 anion exchange resin (80g) absorption, and with 500ml deionized water eluting anion exchange resin, the evaporated under reduced pressure eluent gets extractum 1.0g, wherein alkaloid DNJ mass content 50.2%.
With the Folium Mori filtering residue behind intact alkaloid of said extracted and the flavone component is raw material, adds the 2L deionized water, puts into 60 ℃ of waters bath with thermostatic control, lixiviate 1.5h, extract 2 times, extracted twice liquid is respectively 1.1L and 1.9L, and merge extractive liquid, also filters, regulate filtrate pH4.0, filtrate is concentrated into 400ml behind the 200g activated carbon decolorizing, add ethanol in the extracting solution after concentrating then, and regulating the ethanol volumetric concentration is 60%, leave standstill 12h, centrifugalize must precipitate.Use dehydrated alcohol, acetone and ether washing precipitate successively, get Folium Mori total polysaccharides precipitation 7.1g, wherein neutral polysaccharide content 15%, and acidic polysaccharose content is 56.8%.
Example 3
30% ethanol water that Folium Mori raw material 400g after pulverizing is added 4L, 95 ℃ of reflux, extract, 3 times, each 30min, the extracting solution that obtains three times is respectively 2.8L, 3.5L, 4.2L, and merge extractive liquid, also filters, and filtering residue dries standby, ethanol evaporate to dryness in the gained filtrate is got extractum, in extractum, add the 4L distilled water then, leave standstill 2h, centrifugalize.Supernatant is by the D101 macroporous resin of 200g, 28 ℃ of absorption down.Then earlier with 2L deionized water rinsing macroporous resin, 60% the aqueous acetone solution eluting macroporous resin of reuse 1L, the eluent evaporated under reduced pressure with obtaining obtains extractum 6.5g, and wherein the mass content of total flavones is 37%.
Flow through solution behind the macroporous resin, 001 * 7 * 7 cation exchange resiies by 80g adsorb 5h down at 25 ℃, earlier with 800ml deionized water rinsing resin, with 200ml ammonia eluting resin, collect the ammonia eluent and be concentrated into 50ml again, the eluent after concentrating adsorbs after D201 anion exchange resin (100g), and with 1L deionized water eluting anion exchange resin, the evaporated under reduced pressure eluent gets extractum 1.2g, and wherein the mass content of alkaloid DNJ is 40.3%.
With the Folium Mori filtering residue behind intact alkaloid of said extracted and the flavone component is raw material, adds the 2L deionized water, puts into 80 ℃ of waters bath with thermostatic control, lixiviate 1.5h, extract 2 times, extracted twice liquid is respectively 1.1L and 1.9L, and merge extractive liquid, also filters, regulate filtrate pH2.0, filtrate is concentrated into 500ml behind the 300g activated carbon decolorizing, add ethanol in the extracting solution after concentrating then, and regulating the ethanol volumetric concentration is 15%, leave standstill 12h, centrifugalize must precipitate.Use dehydrated alcohol, acetone and ether washing precipitate successively, get Folium Mori total polysaccharides precipitation 7.0g, wherein neutral polysaccharide content 25%, and acidic polysaccharose content is 59.2%.
Claims (1)
1. the method for a continuously extracting alkaloids from Folium Mori, flavone and polysaccharide active components is characterized in that comprising following process:
1) volume ratio by dry Folium Mori weight and solution is 1: 10～15, it is 20%～90% alcoholic solution that dry Folium Mori after pulverizing are soaked in volumetric concentration, or to be soaked in volumetric concentration be 10%～90% acetone soln, reflux, extract, is 3 times under 95 ℃ of conditions, each 30min that extracts, extracting solution is mixed and filtration under diminished pressure, obtain filtrate and filtering residue;
2) step 1) gained filtrate is removed ethanol through reduction vaporization, volume ratio by dry Folium Mori weight and deionized water is 1: 10～15 adding deionized waters then, left standstill 2 hours, centrifugalize, supernatant 20～30 ℃ of absorption, is at first used ionized water eluting macroporous resin by macroporous resin D4020 or H103, then with ethanol or acetone soln eluting macroporous resin, collect eluent, eluent obtains flavonoid compound extract through evaporation under reduced pressure removed ethanol or acetone;
3) stream is through the solution behind the macroporous resin, be that 001 * 7 or 001 * 7 * 7 cation exchange resin was 20～30 ℃ of absorption 5～10 hours by model, use earlier the deionized water rinsing resin, obtain eluent with ammonia eluting resin again, it is residual that the gained eluent is concentrated to no ammonia, eluent after concentrating is 201 * 7 or 201 * 4 anion exchange resin absorption again by model, reuse deionized water eluting anion exchange resin gets eluent, eluent obtains alkaloid compound extractum through reduction vaporization;
4) the Folium Mori filtering residue that obtains in the step 1) deionized water with 3 times of volumes is decocted twice down at 60～80 ℃, filtration under diminished pressure, collect filtrate, filtrate is through activated carbon decolorizing, and the filtrate decompression after the decolouring is concentrated into 1/5～1/10 of extracting liquid volume, in concentrated solution, add ethanol afterwards, make that the ethanol volumetric concentration is 10%～60% in the concentrated solution, left standstill 10～15 hours, centrifugalize gets precipitate, use dehydrated alcohol, acetone and ether washing precipitate more successively, obtain the matter polysaccharide compound.
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