CN104940609A - Method for extracting and separating multiple active constituents from polygonatum - Google Patents
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Abstract
The invention discloses a method for extracting and separating multiple active constituents from polygonatum. Active constituents including polygahatous polysaccharides, polygahatous saponin and polygahatous flavones are extracted and separated through the homogenate extraction technique. Compared with traditional methods, the method has the advantages that extraction condition is mild, and efficient constituents are prevented from being damaged under heating; solvent consumption is low, extraction is quick and efficient, material extraction can be completely achieved in 0.5-1.5 min, and extraction rate is high; separation and purification are further conducted through macroporous resin, so that purification is effectively improved; wherein, the yield and purity of polygahatous polysaccharides are 28.37% and 88.76% respectively, the yield and purity of polygahatous saponin are 4.95% and 89.49% respectively, and the yield and purity of polygahatous flavones are 1.97% and 82.81% respectively. Polygahatous saponin and polygahatous flavones byproducts are extracted through the homogenate extraction technique so that the polygahatous polysaccharides can be extracted, polygahatous medicinal material resources are saved, the effective extraction rate and comprehensive utilization rate of multiple active constituents of polygonatum are increased, and resource waste is avoided.
Description
Technical field
The invention belongs to the extracting method of Rhizoma Polygonati, belong to the method extracting saponins of rhizoma polygonati, Rhizoma Polygonati total flavones and polygonatum polysaccharide especially.
Background technology
Rhizoma Polygonati is Liliaceae Polygonatum (PolygonatumMill.) perennial root herbaceous plant, calls yellow chicken Lay, Canis familiaris L. Rhizoma Zingiberis Recens etc.In recent years, both at home and abroad the research of HUANGJING ZANYU CAPSULE is deepened continuously, be separated from this platymiscium and obtained the various active compositions such as polysaccharide, saponin, flavone.There are some researches show that polygonatum polysaccharide has and strengthened immunologic function, antitumor, radioprotective, memory reinforcing, fatigue-resisting function; The bioactivity research of sibiricoside focuses mostly in blood sugar lowering, antitumor, AntiHIV1 RT activity etc.; And Rhizoma Polygonati flavone also has biological activity more widely, comprise allaying tiredness, protection blood vessel, atherosclerosis, blood sugar lowering, blood fat reducing, expansion blood capillary, defying age, antibacterial and function such as activation brain etc.Therefore the polysaccharide in extraction and isolation Rhizoma Polygonati, steroidal saponin and flavone isoreactivity composition have important society and economic worth.
The acquisition of polygonatum polysaccharide mainly comprises defat ~ alcohol analyses ~ link such as deproteinization ~ dialysis ~ drying, in extraction process, mainly contain water extraction, weak acid extraction method, weak base extraction method, enzyme lifting manipulation, ultrasound assisted extraction method, microwave―assisted extraction and supercritical fluid extraction etc. at present.The deficiency of the aspects such as above-mentioned polygonatum polysaccharide leaching process life period is long, cost is high, process is complicated, purity of polysaccharide is low.Disclosed in Chinese patent CN 103788224, a kind of method extracting polygonatum polysaccharide from Chinese medicine Rhizoma Polygonati adopts the process route of On Microwave-assisted Extraction ~ concentrated ~ sonic oscillation ~ Deproteinization ~ mistake post, obtaining polygonatum polysaccharide through separation and purification repeatedly, there is the shortcoming of the aspects such as process complexity, the process time is longer, organic solvent consumption is large in this technique.
The acquisition of saponins of rhizoma polygonati mainly comprises the links such as extraction ~ separation ~ purification, in extraction process, mainly contains heating reflux method, soxhlet extraction, supercritical ultrasonics technology, microwave method etc. at present.Disclosed in CN 101837091 patent ~ kind of the continuous process route extracting the technique employing soak extraction ~ extraction ~ upper prop of Rhizoma Polygonati Saponin and polygonatum polysaccharide, be 3.9% of Rhizoma Polygonati gross weight by the said sibiricoside yield of document, for 90.7% of saponins of rhizoma polygonati, purity is 95.3%, but the shortcomings such as in this technique, organic solvent consumption used is large, process time length.
The acquisition of Rhizoma Polygonati flavone mainly comprises 3 key links such as extraction ~ separation ~ purification, in extraction process, mainly contains extraction, cable-styled method, microwave method, circumfluence method etc. at present.Wang Wenjun etc. have published Rhizoma Polygonati extracting flavonoids method, and it adopts defat ~ ultrasound assisted extraction ~ decolouring ~ drying process route, complex technical process, and required organic solvent amount is large, and the time is long, and extraction ratio is on the low side simultaneously.
In recent years, homogenate extraction technology is used to the extraction of Chinese crude drug active component gradually, can greatly reduce extraction time, improves the yield of wherein active component.Because flash extracter operationally, inner edge high-speed rotation with outside generation dissection process, strong vortex can be formed centrally in inner edge, and drive the material inside and outside pulverized to stir, thus produce vigorous stirring effect, the material molecule be extracted in material is exposed to along with diminishing of crushed particles in solvent environment and is also transferred to rapidly in solvent, between Extraction solvent and material particles, the distribution of chemical composition hockets between Balanced or unbalanced fast with the carrying out of fragmentation, finally reach thorough pulverizing, the extraction of complete equipilibrium, and then overflow effective content.
Summary of the invention
Problem solved by the invention is to provide one with Rhizoma Polygonati crude drug for raw material, and extraction process is simple, the time is short, can obtain again the extracting method of high-purity polygonatum polysaccharide, sibiricoside, Rhizoma Polygonati flavone.
The technical scheme that the present invention deals with problems comprises the method for the method extracting saponins of rhizoma polygonati, the method extracting Rhizoma Polygonati total flavones, extraction polygonatum polysaccharide successively.
In Rhizoma Polygonati, a method for various active component extraction separation, comprises the steps:
(1) extraction and isolation of sibiricoside
Mixed with ethanol by SHENGHUANG essence medicinal powder, extract by homogenate extraction technology, sucking filtration, obtains saponins of rhizoma polygonati extracting solution and Rhizoma Polygonati medicinal residues; Ethanol rotary evaporator in saponins of rhizoma polygonati extracting solution is reclaimed, obtains the thick product of saponins of rhizoma polygonati; By thick for saponins of rhizoma polygonati product distilled water diluting, add to ready macroporous resin to adsorb, first carry out eluting with distilled water, till being eluted to the not aobvious Molish reaction of effluent, eluting is carried out again with ethanol, until eluent is considered as eluting completely without saponin, volatilizes the lyophilization of ethanol final vacuum to constant weight, obtain sibiricoside finished product;
(2) extraction and isolation of Rhizoma Polygonati flavone
Mixed with ethanol ~ hydrochloric acid solution by SHENGHUANG essence medicinal powder, extract by homogenate extraction technology, sucking filtration, obtains Rhizoma Polygonati flavone extractive and Rhizoma Polygonati medicinal residues; Ethanol is reclaimed by Rhizoma Polygonati flavone extractive rotary evaporator, obtains the thick product of Rhizoma Polygonati flavone; By thick for Rhizoma Polygonati flavone product distilled water diluting, add to ready macroporous resin to adsorb, first carry out eluting with distilled water, till being eluted to the not aobvious Molish reaction of effluent, eluting is carried out again with ethanol, until eluent is considered as eluting completely without flavone, volatilizes the lyophilization of ethanol final vacuum to constant weight, obtain Rhizoma Polygonati flavone finished product;
(3) extraction and isolation of polygonatum polysaccharide
Merge Rhizoma Polygonati medicinal residues in above-mentioned (1), (2), and add distilled water mixing, extract by homogenate extraction technology, sucking filtration, obtains polysaccharide extraction liquid; Polysaccharide extraction liquid is added to ready macroporous resin to adsorb, and carries out eluting with distilled water, until eluent to be considered as eluting without polysaccharide complete; Polygonatum polysaccharide is obtained by spray-dried for eluent.
The method of various active component extraction separation in above-mentioned Rhizoma Polygonati, preferred step and condition as follows:
(1) extraction and isolation of sibiricoside
A. by Solomonseal Rhizome with 50% ~ 80% ethanol mix (percent by volume), with flash extracter extract 0.5 ~ 1.5min, sucking filtration, obtains Rhizoma Polygonati extracting solution and Rhizoma Polygonati medicinal residues; Described Solomonseal Rhizome is 1:10 ~ 20(grams per liter with the ratio of described ethanol);
B. the ethanol rotary evaporator in Rhizoma Polygonati extracting solution is reclaimed, obtain ethanol extraction; Ethanol extraction is used petroleum ether extraction defat; Extract water-saturated n-butanol after defat is repeatedly extracted, merges n-butanol extracting liquid, after reclaiming n-butyl alcohol, obtain the thick product of saponins of rhizoma polygonati;
C. thick for Rhizoma Polygonati total saponins distilled water being dissolved to concentration is 0.8 ~ 1mg/ml, add to ready macroporous resin to adsorb, first carry out eluting with distilled water, till being eluted to the not aobvious Molish reaction of effluent, be the ethanol (percent by volume) of 70% ~ 80% again by volumetric concentration, carry out eluting, until eluent to be considered as eluting without saponin complete, volatilize the lyophilization of ethanol final vacuum to constant weight, obtain sibiricoside finished product;
(2) extraction and isolation Rhizoma Polygonati flavone
A. by Solomonseal Rhizome with 80% ~ 90% ethanol (percent by volume) mix, with flash extracter extract 0.5 ~ 1.5min, sucking filtration, obtains Rhizoma Polygonati extracting solution and Rhizoma Polygonati medicinal residues;
B. the alcoholic solution rotary evaporator in Rhizoma Polygonati extracting solution is reclaimed, obtain alcoholic solution extract; By the petroleum ether extraction defat of alcoholic solution extract; Extract water-saturated n-butanol after defat is repeatedly extracted, merges n-butanol extracting liquid, after reclaiming n-butyl alcohol, obtain the thick product of Rhizoma Polygonati total flavones;
C. be that 0.8 ~ 1mg/ml adds to ready macroporous resin and adsorbs by thick for Rhizoma Polygonati total flavones product distilled water diluting to concentration, first carry out eluting with distilled water, till being eluted to the not aobvious Molish reaction of effluent, be the ethanol (percent by volume) of 80% ~ 90% again by volumetric concentration, carry out eluting, until eluent is considered as eluting completely without saponin, volatilizes the lyophilization of ethanol final vacuum to constant weight, obtain Rhizoma Polygonati flavone finished product finished product;
(3) extraction and isolation polygonatum polysaccharide
A. above-mentioned Rhizoma Polygonati medicinal residues are merged, and distilled water mixing, extract 0.5 ~ 1.5min with flash extracter, sucking filtration obtains polysaccharide extraction liquid; Described Rhizoma Polygonati medicinal residues are step (1) a and step (2) a gained; Described Rhizoma Polygonati medicinal residues and distilled water mass ratio 1:10 ~ 12 (w/w);
B. polysaccharide extraction liquid is added to ready macroporous resin to adsorb, and carries out eluting with distilled water, until eluent to be considered as eluting without polysaccharide complete; Polygonatum polysaccharide is obtained by spray-dried for eluent.
Compared with prior art, beneficial effect of the present invention shows:
1. the present invention's application sudden strain of a muscle is put forward technology and is extracted various active composition in Rhizoma Polygonati, make extraction time foreshorten to 0.5 ~ 1.5min, and overall process is carried out down all at normal temperatures, effectively prevent the destruction that total saponins in Rhizoma Polygonati, total flavones and active polysaccharide are at high temperature subject to;
2, the inventive method achieves various active composition extraction rapidly and efficiently in Rhizoma Polygonati, and wherein, saponins of rhizoma polygonati yield is 4.95% of Rhizoma Polygonati gross weight, and purity is 89.49%; The yield of Rhizoma Polygonati flavone is 1.97% of Rhizoma Polygonati weight, and purity is 82.81%; The yield of polygonatum polysaccharide is 28.37% of Rhizoma Polygonati weight, and purity is 88.76%; Not only take full advantage of Rhizoma Polygonati resource, improve raw-material use value, and this technology is while extraction total saponins and total flavones, does not affect the extraction of polysaccharide, the response rate is high, low stain (all organic solvents all can reclaim again), the feature such as simple to operate.
3, the organic solvent toxicity that uses of the inventive method is less, is easy to reclaim; And this method has features such as operation is few, equipment is simple, easy to operate, feasibility good, low production cost.
Detailed description of the invention
Below in conjunction with embodiment, invention is described in detail.The percentage ratio of following ethanol is percent by volume.
(1) extraction and isolation of sibiricoside:
Embodiment 1:
Precision takes 4 grams of SHENGHUANG essence medicinal powders and mixes with the ethanol of 40ml50%, and extract 0.5min by homogenate extraction technology, sucking filtration, obtains saponins of rhizoma polygonati extracting solution and Rhizoma Polygonati medicinal residues; Ethanol rotary evaporator in saponins of rhizoma polygonati extracting solution is reclaimed, obtains the thick product of saponins of rhizoma polygonati; By thick for saponins of rhizoma polygonati product distilled water diluting to concentration 0.8mg/ml, add to ready macroporous resin to adsorb, first carry out eluting with distilled water, till being eluted to the not aobvious Molish reaction of effluent, be the ethanol of 70% by volumetric concentration again, carry out eluting, until eluent to be considered as eluting without saponin complete, volatilize the lyophilization of ethanol final vacuum to constant weight, obtain sibiricoside finished product; The yield of gained sibiricoside is 6.85%, and purity is 81.21%.
Embodiment 2:
Precision takes 4 grams of SHENGHUANG essence medicinal powders and mixes with the ethanol of 40ml 65%, and extract 1min by homogenate extraction technology, sucking filtration, obtains saponins of rhizoma polygonati extracting solution and Rhizoma Polygonati medicinal residues; Ethanol rotary evaporator in saponins of rhizoma polygonati extracting solution is reclaimed, obtains the thick product of saponins of rhizoma polygonati; By thick for saponins of rhizoma polygonati product distilled water diluting to concentration 0.9mg/ml, add to ready macroporous resin to adsorb, first carry out eluting with distilled water, till being eluted to the not aobvious Molish reaction of effluent, be the ethanol of 75% by volumetric concentration again, carry out eluting, until eluent to be considered as eluting without saponin complete, volatilize the lyophilization of ethanol final vacuum to constant weight, obtain sibiricoside finished product; The yield of gained sibiricoside is 6.89%, and purity is 79.53%.
Embodiment 3:
Precision takes 4 grams of SHENGHUANG essence medicinal powders and mixes with the ethanol of 40ml 80%, extracts 1.5min, extract at twice, merge extractive liquid, sucking filtration, obtain saponins of rhizoma polygonati extracting solution and Rhizoma Polygonati medicinal residues by homogenate extraction technology; Ethanol rotary evaporator in saponins of rhizoma polygonati extracting solution is reclaimed, obtains the thick product of saponins of rhizoma polygonati; By thick for saponins of rhizoma polygonati product distilled water diluting to concentration 1.0mg/ml, add to ready macroporous resin to adsorb, first carry out eluting with distilled water, till being eluted to the not aobvious Molish reaction of effluent, be the ethanol of 80% by volumetric concentration again, carry out eluting, until eluent to be considered as eluting without saponin complete, volatilize the lyophilization of ethanol final vacuum to constant weight, obtain sibiricoside finished product; The yield of gained sibiricoside is 6.95%, and purity is 79.49%.
(2) extraction and isolation of Rhizoma Polygonati flavone:
Embodiment 4:
Precision takes 4 grams of SHENGHUANG essence medicinal powders and mixes with 30ml 80% ethanol ~ hydrochloric acid solution, and extract 0.5min by homogenate extraction technology, sucking filtration, obtains Rhizoma Polygonati flavone extractive and Rhizoma Polygonati medicinal residues; Ethanol is reclaimed by Rhizoma Polygonati flavone extractive rotary evaporator, obtains the thick product of Rhizoma Polygonati flavone; By thick for Rhizoma Polygonati flavone product distilled water diluting to concentration 0.8mg/ml, add to ready macroporous resin to adsorb, first carry out eluting with distilled water, till being eluted to the not aobvious Molish reaction of effluent, be the ethanol of 80% by volumetric concentration again, carry out eluting, until eluent to be considered as eluting without flavone complete, volatilize the lyophilization of ethanol final vacuum to constant weight, obtain Rhizoma Polygonati flavone finished product; The yield of gained Rhizoma Polygonati flavone is 1.85%, and purity is 83.20%.
Embodiment 5:
Precision takes 4 grams of SHENGHUANG essence medicinal powders and mixes with 30ml 85% ethanol ~ hydrochloric acid solution, and extract 1.0min by homogenate extraction technology, sucking filtration, obtains Rhizoma Polygonati flavone extractive and Rhizoma Polygonati medicinal residues; Ethanol is reclaimed by Rhizoma Polygonati flavone extractive rotary evaporator, obtains the thick product of Rhizoma Polygonati flavone; By thick for Rhizoma Polygonati flavone product distilled water diluting to concentration 0.9mg/ml, add to ready macroporous resin to adsorb, first carry out eluting with distilled water, till being eluted to the not aobvious Molish reaction of effluent, be the ethanol of 85% by volumetric concentration again, carry out eluting, until eluent to be considered as eluting without flavone complete, volatilize the lyophilization of ethanol final vacuum to constant weight, obtain Rhizoma Polygonati flavone finished product; The yield of gained Rhizoma Polygonati flavone is 1.93%, and purity is 82.87%.
Embodiment 6:
Precision takes 4 grams of SHENGHUANG essence medicinal powders and mixes with 30ml 90% ethanol ~ hydrochloric acid solution, and extract 1.5min by homogenate extraction technology, sucking filtration, obtains Rhizoma Polygonati flavone extractive and Rhizoma Polygonati medicinal residues; Ethanol is reclaimed by Rhizoma Polygonati flavone extractive rotary evaporator, obtains the thick product of Rhizoma Polygonati flavone; By thick for Rhizoma Polygonati flavone product distilled water diluting to concentration 1.0mg/ml, add to ready macroporous resin to adsorb, first carry out eluting with distilled water, till being eluted to the not aobvious Molish reaction of effluent, be the ethanol of 85% by volumetric concentration again, carry out eluting, until eluent to be considered as eluting without flavone complete, volatilize the lyophilization of ethanol final vacuum to constant weight, obtain Rhizoma Polygonati flavone finished product; The yield of gained Rhizoma Polygonati flavone is 1.97%, and purity is 82.81%.
(3) polygonatum polysaccharide extraction and isolation:
Embodiment 7:
Merge the Rhizoma Polygonati medicinal residues of above-described embodiment 1 and embodiment 4, and add 80 grams of distilled water mixing, extract 0.5min by homogenate extraction technology, sucking filtration, obtains polysaccharide extraction liquid; Polysaccharide extraction liquid is added to ready macroporous resin to adsorb, and carries out eluting with distilled water, until eluent to be considered as eluting without polysaccharide complete; Polygonatum polysaccharide is obtained by spray-dried for eluent; The yield of gained polygonatum polysaccharide is 28.37%, and the purity of polygonatum polysaccharide is 88.76%.
Embodiment 8:
Merge the Rhizoma Polygonati medicinal residues of above-described embodiment 2 and embodiment 5, and add 80 grams of distilled water mixing, extract 1.0min by homogenate extraction technology, sucking filtration, obtains polysaccharide extraction liquid; Polysaccharide extraction liquid is added to ready macroporous resin to adsorb, and carries out eluting with distilled water, until eluent to be considered as eluting without polysaccharide complete; Polygonatum polysaccharide is obtained by spray-dried for eluent; The yield of gained polygonatum polysaccharide is 28.53%, and the purity of polygonatum polysaccharide is 88.74%.
Embodiment 9:
Merge the Rhizoma Polygonati medicinal residues of above-described embodiment 3 and embodiment 6, and add 80 grams of distilled water mixing, extract 1.5min by homogenate extraction technology, sucking filtration, obtains polysaccharide extraction liquid; Polysaccharide extraction liquid is added to ready macroporous resin to adsorb, and carries out eluting with distilled water, until eluent to be considered as eluting without polysaccharide complete; Polygonatum polysaccharide is obtained by spray-dried for eluent; The yield of gained polygonatum polysaccharide is 28.61%, and the purity of polygonatum polysaccharide is 88.70%.
Embodiment 10:
Get polystyrene type resin (AB-8) 2 grams, use soaked in absolute ethyl alcohol respectively 24 hours, (amount of ethanol is as the criterion with the complete submergence resin of energy), then with ultra-pure water wet method dress post; Rinse resin column with dehydrated alcohol, add 2 times of volume distilled water to effluent and do not produce muddiness, then use ultrapure water resin column extremely without alcohol taste.Then each resin at room temperature vacuum drying, for subsequent use.
Claims (9)
1. the method for various active component extraction separation in Rhizoma Polygonati, is characterized in that, comprise the following steps:
(1) extraction and isolation of sibiricoside:
Mixed with ethanol by SHENGHUANG essence medicinal powder, extract by homogenate extraction technology, sucking filtration, obtains saponins of rhizoma polygonati extracting solution and Rhizoma Polygonati medicinal residues; Ethanol rotary evaporator in saponins of rhizoma polygonati extracting solution is reclaimed, obtains the thick product of saponins of rhizoma polygonati; By thick for saponins of rhizoma polygonati product distilled water diluting, add to ready macroporous resin to adsorb, first carry out eluting with distilled water, till being eluted to the not aobvious Molish reaction of effluent, eluting is carried out again with ethanol, until eluent is considered as eluting completely without saponin, volatilizes the lyophilization of ethanol final vacuum to constant weight, obtain sibiricoside finished product;
(2) extraction and isolation of Rhizoma Polygonati flavone:
Mixed with ethanol ~ hydrochloric acid solution by SHENGHUANG essence medicinal powder, extract by homogenate extraction technology, sucking filtration, obtains Rhizoma Polygonati flavone extractive and Rhizoma Polygonati medicinal residues; Ethanol is reclaimed by Rhizoma Polygonati flavone extractive rotary evaporator, obtains the thick product of Rhizoma Polygonati flavone; By thick for Rhizoma Polygonati flavone product distilled water diluting, add to ready macroporous resin to adsorb, first carry out eluting with distilled water, till being eluted to the not aobvious Molish reaction of effluent, eluting is carried out again with ethanol, until eluent is considered as eluting completely without flavone, volatilizes the lyophilization of ethanol final vacuum to constant weight, obtain Rhizoma Polygonati flavone finished product;
(3) extraction and isolation of polygonatum polysaccharide:
Merge Rhizoma Polygonati medicinal residues in above-mentioned (1), (2), and add distilled water mixing, extract by homogenate extraction technology, sucking filtration, obtains polysaccharide extraction liquid; Polysaccharide extraction liquid is added to ready macroporous resin to adsorb, and carries out eluting with distilled water, until eluent to be considered as eluting without polysaccharide complete; Polygonatum polysaccharide is obtained by spray-dried for eluent.
2. the method for various active component extraction separation in Rhizoma Polygonati according to claim 1, is characterized in that: described SHENGHUANG essence medicinal powder make a living Rhizoma Polygonati medical material through cleaning, dry, pulverize, cross 60 mesh sieves.
3. the method for various active component extraction separation in Rhizoma Polygonati according to claim 1, is characterized in that: the concentration of alcohol in described (1) is 50 ~ 80% percents by volume.
4. the method for various active component extraction separation in Rhizoma Polygonati according to claim 1, is characterized in that: the homogenate extraction time in described (1) is 0.5 ~ 1.5min.
5. the method for various active component extraction separation in Rhizoma Polygonati according to claim 1, it is characterized in that: the ethanol ~ hydrochloric acid solution in described (2) is 4:1 volume ratio, wherein concentration of hydrochloric acid is 25% mass percent.
6. the method for various active component extraction separation in Rhizoma Polygonati according to claim 1, is characterized in that: the concentration of alcohol in said (2) is 80 ~ 90% percents by volume.
7. the method for various active component extraction separation in Rhizoma Polygonati according to claim 1, is characterized in that: the Rhizoma Polygonati medicinal residues in described (3) are the Rhizoma Polygonati medicinal residues in above-mentioned (1) and (2).
8. the method for various active component extraction separation in Rhizoma Polygonati according to claim 1, it is characterized in that: described macroporous resin is polystyrene type, best is AB ~ 8.
9. the method for various active component extraction separation in Rhizoma Polygonati according to claim 1, it is characterized in that: the spray drying condition in described (3) is inlet temperature 120 ~ 170 DEG C, outlet temperature is 75 ~ 90 DEG C, input concentration 15 ~ 25%, and charging rate is 1000 ~ 2000ml/h.
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