CN103524576A - Separation and purification process of 2, 3, 5, 4'-tetrahydroxystilbene-2-O-beta-D-glucoside - Google Patents

Abstract

The present invention provides a separation and purification process for 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside (hereinafter referred to as “stilbene glycoside”). By weighing stilbene The glycoside extract was mixed with silica gel in a loose form, placed on a glass column and eluted with an eluent, tracked and detected by TLC, combined the fractions containing stilbene glycosides, recovered the solvent, and dried to obtain the stilbene glycoside extract. Add pure water to heat to dissolve the stilbene glycoside extract, then freeze and crystallize it, after it dissolves, suction filter and wash with pure water to obtain off-white crystals, transfer the filter residue to a vacuum drying oven for drying, and obtain pure stilbene glycoside ; The process is simple and feasible to operate, and can improve product purity, and can quickly and massively separate and purify stilbene glycosides; the process is easy to operate, low in cost, and stable in yield. After the crystallization treatment, high-purity stilbene glycosides can be obtained, which is suitable for large-scale industrial production.

Description

Separation and purification process of 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside

technical field

The invention relates to the technical field of separation and purification of traditional Chinese medicine, especially the separation and purification process of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside (hereinafter referred to as "stilbene glycoside") . the

Background technique

Polygonum multiflorum is the dry tuberous root of Polygonum multiflorum Thunb., a Polygonum multiflorum Thunb. It was first recorded in "Kaiyuan Materia Medica". It has the effects of eliminating carbuncles, moistening the intestines and laxatives, and also has the functions of nourishing essence and blood, black beard and hair, strengthening muscles and bones, and nourishing the liver and kidney. Pharmacological studies have shown that Polygonum multiflorum has anti-aging, improving the body's non-specific immune function, lowering blood fat, anti-atherosclerosis, protecting the myocardium, protecting the liver, neuroprotection and antibacterial and other pharmacological activities. Clinical research is mainly used for the treatment of hyperlipidemia, insomnia syndrome, white hair, boils, etc. The active ingredients in Radix Polygoni Multiflori mainly include anthraquinones and stilbene glycosides mainly composed of chrysophanol, emodin, rhein, and emodin methyl ether. Among them, anthraquinones can detoxify, eliminate carbuncle, and laxative. The stilbene glycosides have obvious cholesterol-lowering and liver-protecting effects. The stilbene glycosides are the main active ingredients in Polygonum multiflorum. The 2010 edition of the Pharmacopoeia of the People's Republic of China clearly stipulates the total amount of stilbene glycosides. The total amount of stilbene glycosides in Polygonum multiflorum must not be less than 1.0 %. Stilbene glycoside is one of the active ingredients in Polygonum multiflorum with important pharmacological effects. Clinical application and modern pharmacological research have shown that stilbene glycoside has significant anti-aging and lipid-lowering effects. the

How to quickly separate and purify a large amount of stilbene glycosides is a problem of close concern in industrial production. At present, methods for the separation and purification of stilbene glycosides include crystallization, polyamide adsorption chromatography, macroporous resin adsorption chromatography, high-speed countercurrent chromatography, and the like. the

For example, the crystallization method is a relatively traditional extraction method. The coarse powder of Polygonum multiflorum is extracted with 95% ethanol under reflux, and the ethanol is recovered to obtain the liquid extract of Polygonum multiflorum, which is diluted with an appropriate amount of distilled water and extracted three times with an equal amount of ether. Ethyl acetate was refluxed for 5-6 hours, the ethyl acetate layer was collected, concentrated under reduced pressure to 1/10 of the original volume, left to stand at room temperature for crystallization, and filtered to obtain the crude stilbene glycoside. Dissolve the crude product in an appropriate amount of hot water or hot ethyl acetate, filter it while it is hot, let it cool and crystallize, and repeat the crystallization in this way to obtain the refined stilbene glycoside as a gray amorphous crystalline powder. According to HPLC analysis, the purity of stilbene glycosides in the crude product is 68.71%, and that in the fine product is 99.94%. Although the purity of the stilbene glycosides obtained by separation and purification by this method is high, there are still some disadvantages in this method, such as large amount of solvent used, complex process, and low utilization rate of resources. the

In addition, there are researches using high-performance liquid chromatography to separate stilbene glycoside monomers with high purity, but this method has high instrument cost and large solvent consumption, which restricts the development of this technology. There is a need in the art for a method capable of rapidly and/or mass-preparing stilbene glycosides with higher purity. the

Contents of the invention

The technical problem to be solved by the present invention is to provide a separation and purification process for stilbene glycosides. the

For solving the problems of the technologies described above, technical scheme of the present invention is:

The separation and purification process of stilbene glycosides, the specific steps are: accurately weigh the initially purified stilbene glycoside extract, add pure water I to dissolve it, freeze and crystallize it, and after it melts, quickly filter it and wash it with pure water II. Off-white crystals were obtained, and the filter residue was transferred to a vacuum oven for drying to obtain pure stilbene glycosides. the

The preferred separation and purification process of the above-mentioned stilbene glycosides, the weight of the preliminary purified stilbene glycoside extract and the pure water I used to dissolve the extract and the pure water II used to wash the crystals: volume: volume in g /ml/ml is calculated as 1:3-7:3-7, and the freezing and crystallization temperature is -20±1°C-0±1°C. the

The preferred separation and purification process of the above-mentioned stilbene glycosides, the weight of the preliminary purified stilbene glycoside extract and the pure water I used to dissolve the extract and the pure water II used to wash the crystals: volume: volume in g /ml/ml is calculated as 1:3:5, and the freezing and crystallization temperature is -10°C±1°C. the

Preferably, the separation and purification process of the above-mentioned stilbene glycosides, the specific steps are as follows:

(1) Weigh the stilbene glycoside extract, add methanol or 95% ethanol to dissolve it, add silica gel to mix the sample, and grind it into a loose shape after the solvent evaporates. Among them, the stilbene glycoside extract, methanol or 95% The weight/volume/weight ratio of ethanol and silica gel is 100:120-200:100-200 in g/ml/g;

(2) Mix ethyl acetate and silica gel at a volume-to-weight ratio of 5:1 in ml/g, pour into a glass column after removing air bubbles, and continue to add ethyl acetate to wash off the silica gel attached to the tube wall, so that The silica gel on the surface of the glass column is smooth;

(3) Add the sample treated in step (1) to the surface of the glass column, start to elute with ethyl acetate, track and detect by TLC, combine the fractions containing stilbene glycosides, recover the solvent, and dry to obtain stilbene Glycoside extract;

(4) Accurately weigh the preliminarily purified stilbene glycoside extract, add pure water I to dissolve it, and freeze it to crystallize it. After it melts, quickly filter it and wash it with pure water II to obtain off-white crystals. Transfer the filter residue to Dry in a vacuum oven to obtain pure stilbene glycosides. the

Preferably, the separation and purification process of the above-mentioned stilbene glycosides, the weight of the initially purified stilbene glycoside extract in the step (4) and the pure water I used to dissolve the extract and the pure water II used to wash the crystal : Volume: The volume is 1:3-7:3-7 in g/ml/ml, and the freezing and crystallization temperature is -20±1℃-0±1℃. the

Preferably, the separation and purification process of the above-mentioned stilbene glycosides, the weight of the initially purified stilbene glycoside extract in the step (4) and the pure water I used to dissolve the extract and the pure water II used to wash the crystal : Volume: The volume is calculated in g/ml/ml as 1:3:5, and the freezing and crystallization temperature is -10℃±1℃. the

The beneficial effects of the present invention are:

The separation and purification process of the above-mentioned stilbene glycosides overcomes the deficiencies in the existing technology of separating and purifying stilbene glycosides from the extract of stilbene glycosides (polygonum multiflorum extract), the operation is simple and feasible, and the product purity can be improved. Rapidly separate and purify stilbene glycosides in large quantities; the process is simple to operate, pollution-free, low in cost, and stable in yield. While maintaining low cost and high yield, high-purity stilbene can be obtained after recrystallization Stilbene glycosides with good repeatability and wide applicability. Stilbene glycoside samples with a purity of 10%-80% can be purified to more than 96%, which is suitable for large-scale industrial production. the

Description of drawings

Figure 1 shows the content obtained by dissolving in 3ml of pure water, freezing and crystallizing at -10°C±1°C, and washing with 5ml of pure water. the

Figure 2 shows the content obtained by dissolving with 5ml of pure water, freezing and crystallizing at -10°C±1°C, and washing with 5ml of pure water. the

Figure 3 shows the content obtained by dissolving in 7ml of pure water, freezing and crystallizing at -10°C±1°C, and washing with 3ml of pure water. the

Figure 4 shows the content obtained by dissolving with 3ml of pure water, freezing and crystallizing at -10°C±1°C, and washing with 7ml of pure water. the

Figure 5 shows the content obtained by dissolving in 3ml of pure water, freezing and crystallizing at 0°C±1°C, and washing with 3ml of pure water. the

Figure 6 shows the content obtained by dissolving with 5ml of pure water, freezing and crystallizing at 0°C±1°C, and washing with 7ml of pure water. the

Figure 7 shows the content obtained by dissolving in 7ml of pure water, freezing and crystallizing at 0°C±1°C, and washing with 5ml of pure water. the

Figure 8 is the content obtained by dissolving with 7ml of pure water, freezing and crystallizing at -20°C±1°C, and washing with 7ml of pure water. the

Figure 9 is the content obtained by dissolving with 3ml of pure water, freezing and crystallizing at -20°C±1°C, and washing with 5ml of pure water. the

Figure 10 is the content obtained by dissolving with 5ml of pure water, freezing and crystallizing at -20°C±1°C, and washing with 3ml of pure water. the

Fig. 11 is an HPLC detection spectrum for the separation and purification of stilbene glycosides using the extract of stilbene glycosides (50% purity of stilbene glycosides). the

Fig. 12 is an HPLC detection spectrum of the separation and purification of stilbene glycosides with the extract of stilbene glycosides (80%). the

Detailed ways

In order to enable those skilled in the art to better understand the technical solution of the present invention, the technical solution of the present invention will be further described in detail below in conjunction with the drawings and specific embodiments. the

The stilbene glycoside extracts in the following examples are all Polygonum multiflorum extracts purchased from the market, and the purity of the stilbene glycosides in the above stilbene glycoside extracts is 10%-80% as detected by HPLC. The present invention is illustrated by the following examples, but not intended to limit the scope of the present invention. Those skilled in the art can understand that various changes and modifications can be made to the present invention without departing from the spirit and scope of the present invention. The present invention provides general and/or specific descriptions of the materials and test methods used in the tests. While many of the materials and methods of manipulation which are employed for the purposes of the invention are well known in the art, the invention has been described here in as much detail as possible. In the present invention (except Examples 1 and 2, pure stilbene glycosides are detected by ultra-high performance liquid chromatography, chromatographic column: ACQUITY UPLC BEH C18 (3.0×100mm, 1.7 μm); mobile phase: acetonitrile: water=10 : 90; flow rate: 0.4mL/min; column temperature: 50°C; detection wavelength: 320nm), the HPLC method used for the remaining detection of extracts or monomer components is the "Pharmacopoeia of the People's Republic of China" on stilbene glycosides in Polygonum multiflorum The determination method is shown in the table below. The purity was determined by the external standard one-point method. The stilbene glycoside reference substance was purchased from China National Institutes for Food and Drug Control, with a content of 94.7%; batch number: 110844-201109; barcode: 8EOS-NMAB. the

Chromatographic column: Agilent Eclipse XDB-C18 (4.6×150mm, 5μm);

Mobile phase: acetonitrile: water = 18:82

Flow rate: 1mL/min;

Column temperature: 20°C;

Detection wavelength: 320nm;

Method: Isocratic elution with mobile phase (acetonitrile: water = 18:82) for 15 minutes. the

Example 1

Use the extract of stilbene glycosides (50% purity of stilbene glycosides) to separate and purify stilbene glycosides, the specific steps are as follows: 

(1) Weigh 110g of 50% stilbene glycoside extract, put it in a beaker, add 150ml of 95% (v/v) ethanol to fully mix and dissolve it, add 150g of silica gel (200-300 mesh) to mix the sample, and put it in Grind in a fume hood and allow the solvent to evaporate naturally to become loose;

(2) Mix 5000ml of ethyl acetate and 1000g of silica gel (100-200 mesh), stir to remove air bubbles, slowly pour into the glass column after soaking completely, and then continue to add ethyl acetate to remove the Wash the silica gel to make the silica gel on the surface of the glass column smooth;

(3) Slowly add the sample treated in step (1) on the prepared glass column, elute with ethyl acetate, 1 fraction per 600ml, collect 20 fractions in total, and track and detect them by TLC , combined 4-19, blue fluorescence can be observed at 365nm, combined the fractions containing stilbene glycosides, recovered the solvent, and dried to obtain the extract of stilbene glycosides;

(4) Precisely weigh about 70g of the initially purified stilbene glycoside extract, put it in a beaker, heat and dissolve it with about 3 times the amount (210ml) of pure water I, and place it at -10°C±1°C Freeze until it crystallizes and take it out. After melting at room temperature, transfer it to a suction filtration device for suction filtration, and wash it with 5 times the amount (350ml) of pure water II in small amounts. It can be seen that the filter residue is off-white crystals, and the filter residue is transferred to a vacuum. After drying under reduced pressure in a drying oven, 41.78 g of pure stilbene glycosides were obtained. After HPLC detection and low-temperature water treatment, the purity of stilbene glycosides was 97.76%. The results are shown in Figure 11. the

Example 2

Use the extract of stilbene glycosides (80%) to separate and purify stilbene glycosides, the specific steps are as follows:

Weigh about 70g of the stilbene glycoside sample (80% stilbene glycoside extract), put it in a beaker, heat and dissolve it with about 3 times the amount (210ml) of pure water I, and place it in a -10±1°C Freeze until it crystallizes under certain conditions and take it out. After melting at room temperature, transfer it to a suction filtration device for suction filtration, and wash it with 5 times the amount (350ml) of pure water II in small amounts. It can be seen that the filter residue is off-white crystals. Transfer the filter residue After drying in a vacuum oven under reduced pressure, 41.59 g of pure stilbene glycosides were obtained. After HPLC detection and low-temperature water treatment, the purity of stilbene glycosides was 96.50%. The results are shown in Figure 12. the

Example 3

Use the extract of stilbene glycosides (50%) to separate and purify stilbene glycosides, the specific steps are as follows:

(1) Weigh 100g of 50% stilbene glycoside extract, put it in a beaker, add 120ml of 95% ethanol to mix and dissolve it, add 200g of silica gel (200-300 mesh) to mix the sample, grind it in a fume hood and make it The solvent evaporates naturally and becomes loose;

(2) Mix 5000ml of ethyl acetate and 1000g of silica gel (100-200 mesh), stir to remove air bubbles, slowly pour into the glass column after soaking completely, and then continue to add ethyl acetate to remove the Wash the silica gel to make the silica gel on the surface of the glass column smooth;

(3) Slowly add the sample treated in step (1) on the prepared glass column, elute with ethyl acetate, 1 fraction per 600ml, collect 20 fractions in total, and track and detect them by TLC , combined 4-19, blue fluorescence can be observed at 365nm, combined the fractions containing stilbene glycosides, recovered the solvent, and dried to obtain the purified crude product of stilbene glycosides;

(4) Precisely weigh about 10g of the initially purified stilbene glycoside extract, put it in a beaker, heat and dissolve it with about 5 times the amount (50ml) of pure water I, and place it at -20°C±1°C Freeze until it crystallizes and take it out. After melting at room temperature, transfer it to a suction filtration device for suction filtration, and wash it with 5 times the amount (50ml) of pure water II in small amounts. It can be seen that the filter residue is off-white crystals, and the filter residue is transferred to a vacuum. Drying in a drying oven under reduced pressure yielded 7.53 g of pure stilbene glycosides, which were detected by HPLC and treated by low-temperature water-medium method, and the purity of stilbene glycosides was 91.7%. the

Example 4

Use the extract of stilbene glycosides (50%) to separate and purify stilbene glycosides, the specific steps are as follows:

(1) Weigh 100g of 50% stilbene glycoside extract, put it in a beaker, add 200ml of 95% ethanol to mix and dissolve it, add 100g of silica gel (200-300 mesh) to mix the sample, grind it in a fume hood and make it The solvent evaporates naturally and becomes loose;

(2) Mix 5000ml of ethyl acetate and 1000g of silica gel (100-200 mesh), stir to remove air bubbles, slowly pour into the glass column after soaking completely, and then continue to add ethyl acetate to remove the Wash the silica gel to make the silica gel on the surface of the glass column smooth;

(3) Slowly add the sample treated in step (1) on the prepared glass column, elute with ethyl acetate, 1 fraction per 600ml, collect 20 fractions in total, and track and detect them by TLC , combined 4-19, blue fluorescence can be observed at 365nm, combined the fractions containing stilbene glycosides, recovered the solvent, and dried to obtain the purified crude product of stilbene glycosides;

(4) Precisely weigh about 10g of the initially purified stilbene glycoside extract, put it in a beaker, heat and dissolve it with about 4 times the amount (40ml) of pure water I, and place it at -15°C±1°C Freeze until it crystallizes and take it out. After melting at room temperature, transfer it to a suction filtration device for suction filtration, and wash it with 5 times the amount (50ml) of pure water II in small amounts. It can be seen that the filter residue is off-white crystals, and the filter residue is transferred to a vacuum. Drying in a drying oven under reduced pressure yielded 7.47 g of pure stilbene glycosides, which was detected by HPLC and treated by low-temperature water-medium method, and the purity of stilbene glycosides was 94.32%. the

Process comparison test example

Preparation of an extract with 80% stilbene glycoside content: Weigh about 20 g of 50% stilbene glycoside extract, use silica gel column chromatography to elute isocratically with ethyl acetate, and track and detect according to thin-layer chromatography, The stilbene glycosides are completely eluted, the eluents containing the stilbene glycosides are combined, recovered and dissolved to obtain a stilbene glycoside sample powder. The sample powder (about 80% stilbene glycoside content) is used in the following tests. the

Test 1: Weigh about 1.0g of the above-mentioned stilbene glycoside sample into a test tube, heat and dissolve with 3mL pure water, freeze and crystallize it at -10°C±1°C, take it out and wait for it to melt, and use a suction filtration device Suction filter, and wash the filter residue with 5 mL of pure water, and the obtained solid is dried into powder. After HPLC detection, the purity of stilbene glycosides was 96.72%, as shown in Figure 1. the

Test 2: Weigh about 1.0g of the above-mentioned stilbene glycoside sample into a test tube, heat and dissolve it with 5mL pure water, make it freeze and crystallize at -10°C±1°C, take it out and wait for it to melt, and use a suction filtration device Suction filter, and wash the filter residue with 5 mL of pure water, and the obtained solid is dried into powder. After HPLC detection, the purity of stilbene glycosides was 96.31%, as shown in Figure 2. the

Test 3: Weigh about 1.0g of the above-mentioned stilbene glycoside sample into a test tube, heat and dissolve it with 7mL pure water, make it freeze and crystallize at -10°C±1°C, take it out and wait for it to melt, and use a suction filtration device Suction filter, and wash the filter residue with 3 mL of pure water, and the obtained solid is dried into powder. Samples were taken and detected by HPLC. After HPLC detection, the purity of stilbene glycosides was 91.80%, as shown in Figure 3. the

Test 4: Weigh about 1.0 g of the above-mentioned stilbene glycoside sample into a test tube, heat and dissolve with 3 mL of pure water, and make it freeze and crystallize at -10 °C ± 1 °C, take it out and wait for it to dissolve, and use suction filtration The device was suction filtered, and the filter residue was washed with 7 mL of pure water, and the obtained solid was dried into powder. Sampling, after HPLC detection, the purity of stilbene glycosides was 91.31%, as shown in Figure 4. the

Test 5: Weigh about 1.0 g of the above-mentioned stilbene glycoside sample into a test tube, heat and dissolve with 3 mL of pure water, freeze and crystallize it at 0°C±1°C, take it out and wait for it to dissolve, and use a suction filtration device to extract Filter, and wash the filter residue with 3 mL of pure water, and the obtained solid is dried into powder. Sampling, after HPLC detection, the purity of stilbene glycosides was 93.83%, as shown in Figure 5. the

Test 6: Weigh about 1.0 g of the above-mentioned stilbene glycoside sample and place it in a test tube, heat and dissolve it with 5 mL of pure water, make it freeze and crystallize at 0°C±1°C, take it out and wait for it to dissolve, and use a suction filtration device to extract Filter, and wash the filter residue with 7 mL of pure water, and the resulting solid is dried into powder. Sampling, after HPLC detection, the purity of stilbene glycosides was 93.36%, as shown in Figure 6. the

Test 7: Weigh about 1.0 g of the above-mentioned stilbene glycoside sample and place it in a test tube, heat and dissolve it with 7 mL of pure water, make it freeze and crystallize at 0°C±1°C, take it out to be melted, and use a suction filtration device to extract Filter, and wash the filter residue with 5ml of pure water, and the resulting solid is dried into powder. Sampling, after HPLC detection, the purity of stilbene glycosides was 91.51%, as shown in Figure 7. the

Test 8: Weigh about 1.0g of the above-mentioned stilbene glycoside sample into a test tube, heat to dissolve with 7mL of pure water, and make it freeze and crystallize at -20°C±1°C, take it out and wait for it to dissolve, and use suction filtration The device was suction filtered, and the filter residue was washed with 7 mL of pure water, and the obtained solid was dried into powder. Sampling, after HPLC detection, the purity of stilbene glycosides was 93.01%, as shown in Figure 8. the

Test 9: Weigh about 1.0g of the above-mentioned stilbene glycoside sample into a test tube, heat and dissolve it with 3mL pure water, make it freeze and crystallize at -20°C±1°C, take it out and wait for it to dissolve, and use a suction filtration device Suction filter, and wash the filter residue with 5 mL of pure water, and the obtained solid is dried into powder. Sampling, after HPLC detection, the purity of stilbene glycosides was 96.64%, as shown in Figure 9. the

Test 10: Weigh about 1.0 g of the above-mentioned stilbene glycoside sample into a test tube, heat to dissolve with 5 mL of pure water, and make it freeze and crystallize at -20°C±1°C, take it out and wait for it to dissolve, and use suction filtration The device was filtered with suction, and the filter residue was washed with 3 mL of pure water, and the obtained solid was dried into powder. Sampling, after HPLC detection, the purity of stilbene glycosides was 89.25%, as shown in Figure 10. the

In summary, it can be seen from Tests 1-10 that in the separation and purification process of the present invention, the volume of solvent (pure water) used per 1.0 g of stilbene glycoside sample is 3 ml, and the crystallization temperature is -10°C ± 1°C. The effect is better when the volume is 5ml. the

The above detailed description of the separation and purification process of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside with reference to specific embodiments is illustrative rather than limiting , several embodiments can be listed according to the defined scope, so changes and modifications without departing from the general concept of the present invention should fall within the protection scope of the present invention. the

Claims (6)

1. The separation and purification process of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside (hereinafter referred to as "stilbene glycoside") is characterized in that: the specific steps are: precision weighing Measure the initially purified stilbene glycoside extract, add pure water I to dissolve it, and then freeze and crystallize it. After it melts, quickly filter it and wash it with pure water II to obtain off-white crystals. Transfer the filter residue to a vacuum drying oven for drying. Pure stilbene glycosides were obtained. 2. The separation and purification process of stilbene glycoside according to claim 1, characterized in that: the preliminary purified stilbene glycoside extract and pure water I for dissolving the extract and pure water for washing crystals The weight of water II: volume: volume in g/ml/ml is 1:3-7:3-7, and the freezing crystallization temperature is -20±1°C-0±1°C. 3. The separation and purification process of stilbene glycoside according to claim 1 or 2, characterized in that: the preliminary purified stilbene glycoside extract is mixed with pure water I for dissolving the extract and for washing crystals The weight of pure water II: volume: volume is 1:3:5 in terms of g/ml/ml, and the freezing and crystallization temperature is -10°C±1°C. 4. The separation and purification process of stilbene glycoside according to claim 1, characterized in that: the specific steps are as follows: (1) Weigh the stilbene glycoside extract, add methanol or 95% ethanol to dissolve it, add silica gel to mix the sample, and grind it into a loose shape after the solvent evaporates. Among them, the stilbene glycoside extract, methanol or 95% The weight/volume/weight ratio of ethanol and silica gel is 100:120-200:100-200 in g/ml/g; (2) Mix ethyl acetate and silica gel at a volume-to-weight ratio of 5:1 in ml/g, pour into a glass column after removing air bubbles, and continue to add ethyl acetate to wash off the silica gel attached to the tube wall, so that The surface of the glass column is smooth with silica gel; (3) Add the sample treated in step (1) to the surface of the glass column, start to elute with ethyl acetate, track and detect by TLC, combine the fractions containing stilbene glycosides, recover the solvent, and dry to obtain stilbene Glycoside extract; (4) Accurately weigh the preliminarily purified stilbene glycoside extract, add pure water I to dissolve it, and freeze it to crystallize it. After it melts, quickly filter it and wash it with pure water II to obtain off-white crystals. Transfer the filter residue to Dry in a vacuum oven to obtain pure stilbene glycosides. 5. The separation and purification process of stilbene glycoside according to claim 4, characterized in that: the stilbene glycoside extract initially purified in the step (4) is mixed with pure water I for dissolving the extract and The weight of pure water II for washing crystals: volume: volume is 1:3-7:3-7 in g/ml/ml, and the freezing and crystallization temperature is -20±1°C-0±1°C. 6. The separation and purification process of stilbene glycoside according to claim 4 or 5, characterized in that: the stilbene glycoside extract initially purified in the step (4) and the pure water I used to dissolve the extract And the weight of pure water II used for washing crystals: volume: volume is 1:3:5 in terms of g/ml/ml, and the freezing crystallization temperature is -10±1°C.

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