CN108992572B - Extraction and separation method and application of non-polysaccharide components of polygonatum sibiricum - Google Patents
Extraction and separation method and application of non-polysaccharide components of polygonatum sibiricum Download PDFInfo
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Abstract
The invention discloses an extraction and separation method of non-polysaccharide components of polygonatum sibiricum, which comprises the following steps: slicing dried rhizoma Polygonati tuber, adding 95% ethanol, performing first reflux extraction, adding 80% ethanol into the residue obtained by first filtration, performing second reflux extraction, filtering, mixing the filtrates, and concentrating to obtain rhizoma Polygonati crude extract; dispersing the crude extract of rhizoma Polygonati with distilled water, sequentially extracting with petroleum ether, n-butanol, ethyl acetate and dichloromethane to obtain petroleum ether phase, n-butanol phase, ethyl acetate phase and dichloromethane phase, mixing dichloromethane phase and ethyl acetate phase to obtain dichloromethane/ethyl acetate phase, recovering organic solvent to obtain concentrated extract of 3 organic phases, and drying at 60 deg.C to obtain rhizoma Polygonati non-polysaccharide component with optimal antitumor effect of dichloromethane/ethyl acetate phase.
Description
Technical Field
The invention belongs to the technical field of sealwort extraction methods, and particularly relates to an extraction and separation method and application of non-polysaccharide components of sealwort.
Background
Sealwort (Polygonatum sibiricum), also known as: rhizoma Polygonati, herba Elsholtziae Pendulifoliae, rhizoma Gynurae Divaricatae, rhizoma Zingiberis recens, and radix Codonopsis Lanceolatae. Is a plant of Polygonatum, with transverse rhizome, cylindrical shape, enlarged nodule, and no stem. The sealwort is sweet in nature and tasty and refreshing, is a traditional Chinese herbal medicine with homology of medicine and food in China, has fleshy and thick rhizome, contains a large amount of starch, sugar, fat, protein, carotene, vitamins and various other nutritional ingredients, can allay hunger when eaten raw and stewed, has the function of body building, can multiply the vitality of people, has full muscles and strong bone marrow, and is very beneficial to the body. The rhizoma Polygonati polysaccharide has high medicinal value, and has antitumor, antioxidant, immunity regulating, antibacterial, antiinflammatory, blood sugar lowering, and blood lipid reducing effects. The content of polygonatum polysaccharide is an important index for identifying the quality of polygonatum. The application has a very wide market prospect in the aspects of health food, therapeutic drugs, daily skin care products and the like; however, the quality control indexes of polygonatum in the current Chinese pharmacopoeia only have one index of polygonatum polysaccharide content, the characteristic components are undefined and are not systematic enough, so that the quality of polygonatum cannot be comprehensively identified, and the material basis of polygonatum is clarified. At present, many methods for extracting active ingredients of rhizoma polygonati exist, and an alcohol precipitation method is mainly adopted.
For example, patent application No. CN201410306492.7 discloses a polygonatum sibiricum extract, a preparation method and an application thereof, wherein the polygonatum sibiricum extract is prepared by the following steps: after the rhizoma polygonati is subjected to ethanol thermal extraction, the extracting solution is subjected to reduced pressure reflux to obtain a total extract; adding distilled water for suspension, sequentially extracting with petroleum ether, ethyl acetate and n-butanol with equal volume, and recovering solvent under reduced pressure to obtain ethyl acetate extract; the rhizoma polygonati extract has an AGEs (advanced glycation end products) inhibition effect, can be further applied to preparation of medicines for resisting diabetes and complications thereof, and has an important clinical application value. In the application, the polygonatum sibiricum is extracted by ethanol for multiple times, the concentration of the ethanol added each time is constant, the adding sequence of the extracting agent in the extracting process is petroleum ether, ethyl acetate and n-butyl alcohol, the polarity of the extracting agent is changed from small to large, the extraction rate of effective components in the polygonatum sibiricum is low by adopting the extracting method, the components are not separated thoroughly, and particularly polysaccharide components and non-polysaccharide components limit the application of the polygonatum sibiricum.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides an extraction and separation method and application of non-polysaccharide components of polygonatum.
The invention provides a method for extracting and separating non-polysaccharide components of polygonatum sibiricum, which is characterized by comprising the following steps:
1) slicing dried rhizoma Polygonati tubers, adding 95 vol% ethanol water solution, performing first reflux extraction, adding 80 vol% ethanol water solution into the filter residue obtained by first filtration, performing second reflux extraction, filtering, mixing the filtrates, and concentrating to obtain rhizoma Polygonati crude extract;
2) dispersing the crude extract of rhizoma Polygonati with distilled water, sequentially extracting with petroleum ether, n-butanol, ethyl acetate and dichloromethane to obtain petroleum ether phase, n-butanol phase, ethyl acetate phase and dichloromethane phase, mixing dichloromethane phase and ethyl acetate phase to obtain dichloromethane/ethyl acetate phase, recovering organic solvent to obtain concentrated extract of 3 organic phases, and drying at 60 deg.C to obtain rhizoma Polygonati non-polysaccharide component.
Preferably, the mass ratio of the ethanol aqueous solution with the volume concentration of 95% to the rhizoma polygonati tubers added in the first extraction in the step 1) is 5-6: 1.
Preferably, the time for the first reflux extraction in the step 1) is 2-3 h.
Preferably, the mass ratio of the ethanol aqueous solution with the volume concentration of 80% added in the second extraction in the step 1) to the filter residue obtained in the first filtration is 4-5: 1.
Preferably, the time for the second reflux extraction in the step 1) is 1-2 h.
Preferably, the concentration conditions in step 1) are as follows: the temperature is 50-60 deg.C, the vacuum degree is-0.09 MPa, and the specific gravity of the concentrated solution is 1-1.1.
Preferably, the temperature of the two reflux extractions in the step 1) is 80-90 ℃.
The invention also provides the application of the non-polysaccharide components of the sealwort obtained by the extraction and separation method in the aspect of tumor resistance.
The 3 organic phases obtained in the step 2) of the invention, namely the petroleum ether phase, the n-butanol phase and the ethyl acetate/dichloromethane phase, are subjected to organic solvent recovery, and the concentrated solution is dried at 60 ℃, so that the concentrated solution can be used for preparing anti-tumor drugs or health care products.
The extraction method commonly used for the active ingredients of the sealwort at present is an alcohol extraction method, and the active ingredients of the sealwort are extracted by utilizing the fact that the active ingredients are easy to dissolve in ethanol. The polygonatum sibiricum is extracted by ethanol for multiple times, the concentration of the ethanol added each time is constant, the obtained extracting solution is added with an extracting agent for extraction, the adding sequence of the extracting agent is from small to large according to the polarity (patent application No. CN201410306492.7), the extraction method can lead the extraction rate of effective components in the polygonatum sibiricum to be lower, and the components are not separated completely, especially polysaccharide components and non-polysaccharide components, so the application of the polygonatum sibiricum is limited.
According to the invention, the polygonatum rhizome tubers are respectively subjected to two times of reflux extraction by using an ethanol water solution with the volume concentration of 95% and an ethanol water solution with the volume concentration of 80%, and the inventor discovers that the concentration of ethanol added in the two times of reflux extraction is different, and only if the volume concentration of ethanol added in the two times is respectively 95% and 80%, the non-polysaccharide components in the polygonatum rhizome can be extracted to the maximum extent, so that the effective utilization of the polygonatum rhizome is realized. In addition, the extracting solution obtained by extraction is added with an extracting agent for extraction, firstly, petroleum ether is added for degreasing the sealwort component, and then, n-butanol, ethyl acetate and dichloromethane are sequentially used for further extraction to obtain four organic phases and a water phase; the invention breaks the adding sequence of the conventional extracting agent (namely, the adding sequence is from small to large according to the polarity), and the effective components in the sealwort can be fully separated according to the adding sequence of the extracting agent of the invention and the concentration of ethanol during reflux extraction, especially the non-polysaccharide components, so that the high-efficiency utilization of the sealwort in various medicament aspects is realized, and the application range of the sealwort is expanded.
The petroleum ether phase, the n-butanol phase, the ethyl acetate/dichloromethane phase and the water phase which are finally extracted are subjected to anti-tumor research, and the anti-tumor effect of the ethyl acetate/dichloromethane phase is found to be the best.
Separating and purifying ethyl acetate/dichloromethane phase by repeated column chromatography, preparative high performance liquid chromatography and glucose gel chromatography, and separating to obtain 10 compounds including 4 flavones, 2 alkaloids, 1 steroid saponin, 1 daucosterol and 2 fatty acids.
The invention has the beneficial effects that:
1. according to the invention, the polygonatum rhizome tubers are respectively subjected to two times of reflux extraction by using an ethanol water solution with the volume concentration of 95% and an ethanol water solution with the volume concentration of 80%, and the inventor discovers that the concentration of ethanol added in the two times of reflux extraction is different, and only if the concentration of ethanol added in the two times is respectively 95% and 80%, the non-polysaccharide components in the polygonatum rhizome can be extracted to the maximum extent, so that the effective utilization of the polygonatum rhizome is realized.
2. The invention breaks the adding sequence of the conventional extracting agent (namely, the adding sequence is from small to large according to the polarity), and the effective components in the sealwort can be fully separated according to the adding sequence of the extracting agent of the invention and the concentration of ethanol during reflux extraction, particularly polysaccharide components and non-polysaccharide components, so that the high-efficiency utilization of the sealwort in various medicament aspects is realized, and the application range of the sealwort is expanded.
3. The preparation method is simple and easy to operate, the yield of the non-polysaccharide components of the polygonatum is high, the polygonatum resource is fully utilized, the use value of the raw materials is improved, the extraction of the polysaccharide is not influenced while the non-polysaccharide is extracted by the technology, the recovery rate is high, and the pollution is low (all organic solvents can be recovered again).
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to specific embodiments below.
Example 1
1) Slicing dried rhizoma polygonati tubers, adding an ethanol water solution with the volume concentration of 95%, wherein the mass ratio of the ethanol water solution to the rhizoma polygonati tubers is 5:1, carrying out first reflux extraction at 80 ℃, after 2h, adding an ethanol water solution with the volume concentration of 80% into filter residues obtained by first filtration, wherein the mass ratio of the 80% ethanol water solution to the filter residues obtained by first filtration is 5:1, carrying out second reflux extraction at 80 ℃, filtering after 1h, combining filtrates obtained by two times of extraction, and concentrating to obtain rhizoma polygonati crude extract, wherein the concentration conditions are as follows: the temperature is 50 ℃, the vacuum degree is-0.09 MPa, and the specific gravity of the concentrated solution is 1;
2) adding distilled water with the mass of 2 times to the crude polygonatum sibiricum extract for dispersing, then sequentially extracting by using petroleum ether, n-butanol, ethyl acetate and dichloromethane, obtaining a petroleum ether phase, an n-butanol phase, an ethyl acetate phase, a dichloromethane phase and a water phase after extraction is finished, combining the dichloromethane phase and the ethyl acetate phase to obtain a dichloromethane/ethyl acetate phase, finally extracting non-polysaccharide components in the polygonatum sibiricum to 3 organic phases, namely the petroleum ether phase, the n-butanol phase and the ethyl acetate/dichloromethane phase, recovering organic solvents in the organic phases to obtain concentrated extracts of the 3 organic phases, drying the concentrated extracts at 60 ℃ to obtain non-polysaccharide components of the polygonatum sibiricum, and finally obtaining the non-polysaccharide component of the polygonatum sibiricum with the extraction rate of 90.54%.
Example 2
1) Slicing dried rhizoma polygonati tubers, adding an ethanol water solution with the volume concentration of 95%, wherein the mass ratio of the 95% ethanol water solution to the rhizoma polygonati tubers is 6:1, carrying out first reflux extraction at 88 ℃, after 3h, adding an ethanol water solution with the volume concentration of 80% into filter residues obtained by first filtration, wherein the mass ratio of the 80% ethanol water solution to the filter residues obtained by first filtration is 5:1, carrying out second reflux extraction at 88 ℃, filtering after 2h, combining filtrates obtained by two extractions, and concentrating to obtain rhizoma polygonati crude extract, wherein the concentration conditions are as follows: the temperature is 55 ℃, the vacuum degree is-0.09 MPa, and the specific gravity of the concentrated solution is 1.06;
2) adding distilled water with the mass of 2 times to the crude polygonatum sibiricum extract for dispersing, then sequentially extracting by using petroleum ether, n-butanol, ethyl acetate and dichloromethane, obtaining a petroleum ether phase, an n-butanol phase, an ethyl acetate phase, a dichloromethane phase and a water phase after extraction is finished, combining the dichloromethane phase and the ethyl acetate phase to obtain a dichloromethane/ethyl acetate phase, finally extracting non-polysaccharide components in the polygonatum sibiricum to 3 organic phases, namely the petroleum ether phase, the n-butanol phase and the ethyl acetate/dichloromethane phase, recovering organic solvents in the organic phases to obtain concentrated extracts of the 3 organic phases, drying the concentrated extracts at 60 ℃ to obtain non-polysaccharide components of the polygonatum sibiricum, and finally obtaining the non-polysaccharide component of the polygonatum sibiricum with the extraction rate of 93.68%.
Example 3
1) Slicing dried rhizoma polygonati tubers, adding an ethanol water solution with the volume concentration of 95%, wherein the mass ratio of the ethanol water solution to the rhizoma polygonati tubers is 5.5:1, carrying out first reflux extraction at 90 ℃, after 2.5h, adding an ethanol water solution with the volume concentration of 80% into filter residues obtained by first filtration, wherein the mass ratio of the 80% ethanol water solution to the filter residues obtained by first filtration is 4:1, carrying out second reflux extraction at 90 ℃, filtering after 1.5h, combining filtrates obtained by two extractions, and concentrating to obtain rhizoma polygonati crude extract, wherein the concentration conditions are as follows: the temperature is 60 ℃, the vacuum degree is-0.09 MPa, and the specific gravity of the concentrated solution is 1.1;
2) adding distilled water with the mass of 2 times to the crude polygonatum sibiricum extract for dispersing, then sequentially extracting by using petroleum ether, n-butanol, ethyl acetate and dichloromethane, obtaining a petroleum ether phase, an n-butanol phase, an ethyl acetate phase, a dichloromethane phase and a water phase after extraction is finished, combining the dichloromethane phase and the ethyl acetate phase to obtain a dichloromethane/ethyl acetate phase, finally extracting non-polysaccharide components in the polygonatum sibiricum to 3 organic phases, namely the petroleum ether phase, the n-butanol phase and the ethyl acetate/dichloromethane phase, recovering organic solvents in the organic phases to obtain concentrated extracts of the 3 organic phases, drying the concentrated extracts at 60 ℃ to obtain non-polysaccharide components of the polygonatum sibiricum, and finally obtaining the non-polysaccharide component of the polygonatum sibiricum with the extraction rate of 91.22%.
Comparative example 1
1) Slicing dried rhizoma polygonati tubers, adding an ethanol water solution with the volume concentration of 80%, wherein the mass ratio of the 80% ethanol water solution to the rhizoma polygonati tubers is 6:1, carrying out first reflux extraction at 88 ℃, after 3h, adding the 80% ethanol water solution into filter residues obtained by first filtration, wherein the mass ratio of the 80% ethanol water solution to the filter residues obtained by first filtration is 5:1, carrying out second reflux extraction at 88 ℃, filtering after 2h, combining filtrates obtained by two extractions, and concentrating to obtain rhizoma polygonati crude extract, wherein the concentrating conditions are as follows: the temperature is 55 ℃, the vacuum degree is-0.09 MPa, and the specific gravity of the concentrated solution is 1.06;
2) adding distilled water with the mass of 2 times to the crude polygonatum sibiricum extract for dispersing, then sequentially extracting by using petroleum ether, n-butanol, ethyl acetate and dichloromethane, obtaining a petroleum ether phase, an n-butanol phase, an ethyl acetate phase, a dichloromethane phase and a water phase after extraction is finished, combining the dichloromethane phase and the ethyl acetate phase to obtain a dichloromethane/ethyl acetate phase, finally extracting non-polysaccharide components in the polygonatum sibiricum to 3 organic phases, namely the petroleum ether phase, the n-butanol phase and the ethyl acetate/dichloromethane phase, recovering organic solvents in the organic phases to obtain concentrated extracts of the 3 organic phases, drying the concentrated extracts at 60 ℃ to obtain non-polysaccharide components of the polygonatum sibiricum, and finally obtaining the non-polysaccharide component of the polygonatum sibiricum with the extraction rate of 78.31%.
Comparative example 2
1) Slicing dried rhizoma polygonati tubers, adding an ethanol water solution with the volume concentration of 95%, wherein the mass ratio of the 95% ethanol water solution to the rhizoma polygonati tubers is 6:1, carrying out first reflux extraction at 88 ℃, after 3h, adding an ethanol water solution with the volume concentration of 80% into filter residues obtained by first filtration, wherein the mass ratio of the 80% ethanol water solution to the filter residues obtained by first filtration is 5:1, carrying out second reflux extraction at 88 ℃, filtering after 2h, combining filtrates obtained by two extractions, and concentrating to obtain rhizoma polygonati crude extract, wherein the concentration conditions are as follows: the temperature is 55 ℃, the vacuum degree is-0.09 MPa, and the specific gravity of the concentrated solution is 1.06;
2) adding distilled water with the mass of 2 times to the crude polygonatum sibiricum extract for dispersing, then sequentially extracting by using petroleum ether, n-butanol, ethyl acetate and dichloromethane to obtain a petroleum ether phase, a dichloromethane phase, an ethyl acetate phase, an n-butanol phase and a water phase after extraction is finished, combining the dichloromethane phase and the ethyl acetate phase to obtain a dichloromethane/ethyl acetate phase, finally extracting non-polysaccharide components in the polygonatum sibiricum to 3 organic phases, namely the petroleum ether phase, the ethyl acetate/dichloromethane phase and the n-butanol phase, recovering organic solvents in the organic phases to obtain concentrated extracts of the 3 organic phases, and drying the concentrated extracts at 60 ℃ to obtain non-polysaccharide components of the polygonatum sibiricum, wherein the extraction rate of the non-polysaccharide of the polygonatum sibiricum is 85.41 finally.
The extraction rate of the non-polysaccharide components of the sealwort in the embodiments 1-3 of the invention is more than 90 percent, which is higher than that in the comparative examples 1-2, and the extraction method of the invention is adopted to fully separate the non-polysaccharide components of the sealwort, thus realizing the high-efficiency utilization of the sealwort in various medicament aspects and expanding the application range of the sealwort.
Research on antitumor activity of each extract phase of rhizoma polygonati
1. In vitro antitumor assay
The petroleum ether phase, the dichloromethane/ethyl acetate phase, the n-butanol phase and the aqueous phase obtained in example 2 were subjected to in vitro antitumor experiments by recovering the reagents, drying the reagents at 60 ℃ respectively and drying the samples.
1.1 construction of cell culture and related cell library platforms
TABLE 1 cell lines used in the experiments
1.2 Experimental part
1.2.1MTT assay to analyze the Effect of different extractions on tumor cell proliferation
The experimental principle is as follows: succinate dehydrogenase exists on the mitochondrial inner membrane of the living cells, yellow-green thiazole blue (MTT for short, which is a dye accepting hydrogen ions) can be degraded into blue-purple formazan by the succinate dehydrogenase, the more the living cells are, the more blue-purple formazan is generated, and the dead cells have no function because the succinate dehydrogenase activity on the mitochondrial inner membrane disappears. The blue-violet formazan is dissolved by using dimethyl sulfoxide, and the absorbance value is measured at 490nm wavelength by using an enzyme labeling instrument, so that the number of living cells can be quantitatively reacted.
1.2.2 different extract phases inhibition HepG-2 and MCF-7 cell proliferation experiments
(1) Grouping experiments: the experiment was divided into aqueous phase, n-butanol phase, dichloromethane/ethyl acetate phase, petroleum ether phase and control. Cyclophosphamide is used as a positive control group; each group is provided with 3 multiple holes.
(2) Cell culture: taking HepG-2 and MCF-7 cells in logarithmic phase, sucking out original culture solution, washing twice with sterile PBS, adding a proper amount of pancreatin for digestion, and centrifugally collecting cells to prepare cell suspension; cells were counted and diluted to 7x10 cell concentration4Perml, sterile 96-well cell culture plates were prepared and labeled accordingly, and 100. mu.l of cell suspension was added to each well, and the mixture was incubated at 37 ℃ with 5% CO2Culturing in an incubator. When the cells grow to about 80% adherent, corresponding drugs are added into each hole of the experimental group, the experimental group is placed at 37 ℃ and 5% CO2Culturing in an incubator for 48 h. The original culture was aspirated, washed twice with sterile PBS, and then 0.5mg/mL M was added to each wellTT culture medium was continued for 4 h. After 4h, the medium in the wells is carefully aspirated, 150. mu.l of dimethyl sulfoxide is added to each well, and the mixture is placed on a shaking table and shaken at a constant speed for 10min to fully dissolve crystals. And measuring the absorbance values of all groups at 490nm by using an enzyme-labeling instrument, and calculating the cell proliferation inhibition rate.
1.3 results and analysis
1.3.1MTT test results
After different extraction phases are applied to HepG-2 and MCF-7 cells for 24 hours, the absorbance of each hole is measured by an enzyme-labeling instrument, and the cell proliferation inhibition rate is calculated, wherein the result is shown in Table 2, wherein IC50 is the half inhibition rate.
TABLE 2 inhibition of different extracts against tumor cell lines
As can be seen from the data in Table 2, different extracts of the present invention have strong inhibitory effects on various tumor cells of breast cancer and liver cancer, and show very good anti-tumor efficacy, wherein the anti-tumor effect of the dichloromethane/ethyl acetate phase is the best, and is better than that of the aqueous phase, and unexpected technical effects are obtained.
2. In vivo antitumor assay
The petroleum ether phase, the dichloromethane/ethyl acetate phase, the n-butanol phase and the aqueous phase obtained in example 2 were subjected to in vivo antitumor experiments by recovering the reagents, drying at 60 ℃ respectively, and drying the samples.
2.1 Experimental materials
The experiment comprises a water phase, a n-butanol phase, a dichloromethane/ethyl acetate phase, a petroleum ether phase and a control group, wherein cyclophosphamide is used as a positive control group.
2.2 Experimental methods
Experimental animals: the Kunming mouse is taken, the weight of the Kunming mouse is 18-22 g, and the Kunming mouse can be used for both male and female. Collecting lung cancer Lewis tumor mass inoculated for 10 days to prepare Lewis cell suspension (cell number 1 × 10)7/ml), except for the healthy control group, the right axilla of the upper limb of the other mice are inoculated with 0.2ml of lung cancer Lewis cell suspension subcutaneously, and after 24 hours of inoculation, the mice are randomly divided into physiological saline groups20ml/kg, 3g/kg for the experimental group of dried samples of petroleum ether phase, dichloromethane/ethyl acetate phase, n-butanol phase and aqueous phase, respectively, and 60mg/kg for the control group of Cyclophosphamide (CTX) for intraperitoneal injection. Each group was administered for 14 days, all mice were sacrificed at day 15, tumor masses were weighed, and tumor inhibition rates were calculated. The results of the specific experiments are shown in table 3.
TABLE 3 inhibition of Lewis of different extracts against mouse lung carcinoma
Note: p <0.05 compared to saline group; p < 0.01.
As can be seen from the in vivo anti-tumor data shown in Table 3, the petroleum ether phase, the dichloromethane/ethyl acetate phase, the n-butanol phase and the water phase obtained by the method can effectively inhibit the growth of the Lewis lung cancer of the mice, wherein the dichloromethane/ethyl acetate phase has the best anti-tumor effect and is superior to the water phase, and an unexpected technical effect is achieved.
Claims (2)
1. A method for extracting and separating non-polysaccharide components from rhizoma polygonati is characterized by comprising the following steps:
1) slicing dried rhizoma polygonati tubers, adding an ethanol water solution with the volume concentration of 95%, performing first reflux extraction, wherein the mass ratio of the ethanol water solution with the volume concentration of 95% to the rhizoma polygonati tubers is 5-6:1 during the first reflux extraction, the time for the first reflux extraction is 2-3h, adding an ethanol water solution with the volume concentration of 80% into filter residues obtained by first filtration, performing second reflux extraction, wherein the mass ratio of the ethanol water solution with the volume concentration of 80% to the filter residues obtained by the first filtration is 4-5:1 during the second reflux extraction, and the time for the second reflux extraction is 1-2h, filtering, combining filtrates obtained by two times of extraction, and concentrating to obtain a rhizoma polygonati crude extract;
2) dispersing the crude extract of rhizoma Polygonati with distilled water, sequentially extracting with petroleum ether, n-butanol, ethyl acetate and dichloromethane to obtain petroleum ether phase, n-butanol phase, ethyl acetate phase and dichloromethane phase, mixing dichloromethane phase and ethyl acetate phase to obtain dichloromethane/ethyl acetate phase, recovering organic solvent to obtain concentrated extract of 3 organic phases, and drying at 60 deg.C to obtain rhizoma Polygonati non-polysaccharide component;
the concentration conditions in the step 1) are as follows: the temperature is 50-60 deg.C, the vacuum degree is-0.09 MPa, the specific gravity of the concentrated solution is 1-1.1, and the temperature of two reflux extractions is 80-90 deg.C.
2. The use of the non-polysaccharide fraction of Polygonati officinalis rhizoma of claim 1 in the preparation of an anti-tumor medicament.
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| CN104069348A (en) * | 2014-06-30 | 2014-10-01 | 天津中医药大学 | Sealwort extract as well as preparation method and use thereof |
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| WO2006099804A1 (en) * | 2005-03-23 | 2006-09-28 | Lee's Pharmaceutical (Hong Kong) Limited | Herbal compositions useful in cancer treatment |
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