CN111686138B - Duckweed extract and preparation method and application thereof - Google Patents
Duckweed extract and preparation method and application thereof Download PDFInfo
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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Abstract
The invention discloses a duckweed extract, which comprises four caffeoyl quinic acid compounds: 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid, and 3,4, 5-O-tricaffeoylquinic acid; in the duckweed extract, the mass content of the four caffeoylquinic acid compounds is 5% or more. The invention also discloses a preparation method of the duckweed extract and application of the duckweed extract in preparing medicines, cosmetics or medical devices with the effect of resisting radiation damage. Biological activity determination proves that the duckweed extract can effectively prevent cell damage caused by radiation, and the further obtained extract refined product also has the same radiation damage resistance effect. The duckweed extract and the refined product of the extract obtained by further purifying the duckweed extract can be used as raw materials for preparing medicines, cosmetics or medical instruments with the radiation damage resistance.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a duckweed extract, a preparation method thereof and application thereof in preparing medicines, cosmetics or medical instruments with the function of resisting radiation damage.
Background
In today's society, radiation has become an important factor affecting human health. In daily life, microwaves, mobile phones, computers, televisions and the like are increasingly popularized, almost all radiation is available, and the influence on human health is increasingly wide and lasting. In medical diagnosis and treatment, radioactive diagnosis and treatment are also widely used, and especially for tumor patients, radiotherapy is a common method. After a human body is irradiated by a certain dose, radiation sickness symptoms of different degrees can be generated, and tissues with vigorous metabolism, such as bone marrow, gonads, immune systems and the like, are easy to damage and can cause death seriously. Even at low doses, prolonged irradiation can cause various radiation disorders, such as hemogram and hematopoietic changes, decreased fertility, cataracts, hair loss, premature aging, malignancies, and the like. At present, some chemical drugs with anti-radiation effect have high toxicity, so that the search for high-efficiency and low-toxicity plant extracts is further applied to drug development and has important significance.
The duckweed is the whole plant of Azolla imbricata (also called Azolla imbricata) and Azolla immbricate (Roxb.) Nakai, and has the effects of relieving exterior syndrome, promoting eruption, inducing sweat, and inducing diuresis. The patent CN102727597A and CN1726939A disclose that the medicine composition containing the duckweed has the effect of resisting radiation injury, but the radiation-resisting effect of the duckweed single medicine is not examined.
The duckweed mainly contains compounds such as chlorogenic acids (including caffeoyl quinic acids), flavonoids, coumarins, cinnamoyl tyrosine derivatives, lignins and fatty acids (3.J.Chromatogr.A,2019,1609,460435; 4.J.Pharm.biomed.anal.,2019,163, 197-203; 5.bot.Mag.Tokyo.1982,95, 303-308; 6.J.Soc.Cosmet.scientists Korea,2010,36, 71-77). However, until now, researches on the basis of the anti-radiation drug effect substances of the duckweed are not found, and duckweed extracts and reports on the prevention and treatment of radiation damage of the duckweed are not found.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a duckweed extract, a preparation method thereof and application thereof in preparing medicines, cosmetics or medical devices with the effect of resisting radiation damage.
The first purpose of the invention is to provide a duckweed extract, which comprises four caffeoyl quinic acid compounds: 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid, and 3,4, 5-O-tricaffeoylquinic acid; in the duckweed extract, the mass content of the four caffeoylquinic acid compounds is 5% or more.
The second purpose of the invention is to provide a refined duckweed extract product, which comprises four caffeoyl quinic acid compounds: 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid, and 3,4, 5-O-tricaffeoylquinic acid; in the duckweed extract, the mass content of the four caffeoylquinic acid compounds is more than 30%.
The third purpose of the invention is to provide the preparation method of the duckweed extract, which comprises the following steps:
step (1), extraction step: pulverizing dried whole herba Spirodelae, extracting with extractant to obtain primary extractive solution, concentrating, centrifuging or filtering, and collecting supernatant or filtrate;
step (2), a purification step: and (2) carrying out macroporous resin adsorption on the supernatant or filtrate obtained in the step (1), sequentially eluting with water and a 5-30% lower alcohol aqueous solution to remove impurities, eluting with a 30-95% lower alcohol aqueous solution, concentrating the eluate to dryness or drying after removing lower alcohol to obtain the duckweed extract.
In a preferred embodiment of the present invention, in the duckweed extract obtained after the steps (1) and (2), four caffeoylquinic acid compounds: the weight content of 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid and 3,4, 5-O-tricaffeoylquinic acid is above 5%.
As a preferred embodiment of the present invention, the preparation method further comprises the step of further purifying: and (3) further purifying the mass content of the four caffeoylquinic acids in the duckweed extract obtained in the step (2) to more than 30% by adopting a method comprising solvent extraction, gel column chromatography, ODS column chromatography and liquid phase preparation to obtain a refined duckweed extract product.
As a preferred embodiment of the present invention, the extractant in step (1) is at least one of water, ethanol and methanol.
As a preferred embodiment of the present invention, the lower alcohol in the step (2) is C 1 -C 5 Is of low orderAn alcohol.
The analytical method of the chemical components of the duckweed extract comprises the following steps:
performing column chromatography on the duckweed extract obtained in the step (2) by adopting MCI, gel, ODS and the like and performing equal separation on a semi-prepared liquid phase to obtain four main components, and identifying the four main components into four caffeoyl quinic acid compounds by spectrum methods such as NMR, MS and the like: 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid, and 3,4, 5-O-tricaffeoylquinic acid. Quantitative analysis by an HPLC-UV method shows that the weight content of the four caffeoylquinic acid compounds in the duckweed extract is more than 5%. Further purifying by solvent extraction, gel column chromatography, ODS column chromatography, preparation liquid phase and other methods to obtain refined duckweed extract products with different purities, wherein the mass content of the four caffeoylquinic acid compounds is more than 30%.
The fourth purpose of the invention is to provide the application of the duckweed extract or the refined duckweed extract product in preparing medicines, cosmetics or medical devices for resisting radiation damage.
The fifth object of the present invention is to provide a pharmaceutical, cosmetic or medical device having an effect of protecting against radiation damage, comprising the duckweed extract or the refined duckweed extract as described above.
Compared with the prior art, the invention has the beneficial effects that:
biological activity tests prove that the duckweed extract can effectively prevent cell damage caused by radiation, and the further obtained refined extract product also has the same radiation damage resistance effect. The duckweed extract and the refined extract product obtained by further purifying the duckweed extract can be used as raw materials for preparing medicines, cosmetics or medical instruments with the function of resisting radiation damage.
Drawings
Fig. 1, fig. 2 and fig. 3 are graphs of experimental data GraphPad of the anti-radiation activity of duckweed extract a (fig. 1), B (fig. 2) and C (fig. 3), respectively, on in vitro cells according to the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1: preparation method of herba Spirodelae extract and refined product thereof
10 kilograms of dried duckweed whole plant is crushed, soaked in 50L of water for 12 hours, added with 50L of water and boiled for 2 times, each time lasts for 2 hours, the extracting solution is combined and concentrated to 50L, then the filtering solution is filtered, the filtrate passes through a 20L HP20 type macroporous resin column at the flow rate of 2BV/h (BV is the volume of a column bed of the macroporous resin), 100L of distilled water is used for leaching to remove impurities, 80L of 60 percent ethanol is used for elution, the eluent is collected and concentrated to be dry, 64g of the duckweed extract is obtained, and the yield is 0.6 percent based on crude drugs. HPLC analysis shows that the sum of the mass contents of the four caffeoylquinic acid compounds in the duckweed extract is 5.6%. The extract is further purified by gel column to obtain refined herba Spirodelae extract with total content of four caffeoylquinic acid compounds of 43 wt%.
Example 2: preparation method of herba Spirodelae extract and its refined product
10 kilograms of dried duckweed whole plant is crushed, soaked in 50L of 50% ethanol for 12 hours and then percolated, 100L of percolate is collected and concentrated to 20L, then filtered, added with water to be regulated to 50L, the filtrate passes through a 20L HZ818 type macroporous resin column at the flow rate of 2BV/h (BV is the volume of a column bed of the macroporous resin), 100L of distilled water is used for leaching to remove impurities, 80L of 70% ethanol is used for elution, the eluent is collected and concentrated to be dry, and then the extract 353g of the duckweed is obtained, wherein the yield is 3.5 percent based on crude drugs. HPLC analysis shows that the sum of the mass contents of the four caffeoylquinic acid compounds in the duckweed extract is 13%. The extract is further purified by ODS column to obtain refined herba Spirodelae extract with total content of four caffeoylquinic acid compounds of 32%.
Example 3: preparation method of herba Spirodelae extract and its refined product
10 kilograms of dried duckweed whole plant is crushed, soaked for 1 hour by 200L of 70% ethanol, extracted for 2 minutes by a flash extractor, the extract is concentrated to 20L, then filtered, added with water and adjusted to 100L, the filtrate passes through a 20L NKA-2 type macroporous resin column at the flow rate of 2BV/h (BV is the volume of a column bed of the macroporous resin), 100L of distilled water is used for leaching to remove impurities, 80L of 80% ethanol is used for elution, the eluent is collected and concentrated to be dry, 404g of the duckweed extract is obtained, and the yield is 4.0 percent based on crude drugs. The sum of the mass contents of the four caffeoylquinic acid compounds in the duckweed extract is 11% by HPLC analysis. The extract is further extracted by a solvent and purified by gel column chromatography to obtain a refined duckweed extract, wherein the sum of the mass contents of the four caffeoylquinic acid compounds in the duckweed extract is 55%.
Example 4: determination of in vitro cell anti-radiation activity of duckweed extract and refined product thereof
First, screening the range of the dose of the extract without toxic side effects on the cells with human normal colon epithelial cells (NCM 460); then, after the cells were pretreated with the extract, NCM460 cells were irradiated and the change in cell activity was observed.
1. Safety detection of the duckweed extract:
taking NCM460 cells in logarithmic growth phase at a cell density of 1 × 10 4 one/mL cell suspension was seeded in 96-well plates at 100. mu.L/well. NCM460 cells were treated at different doses for each sample and tested for toxic side effects of the drug in the absence of radiation using the MTT assay. The results show that the duckweed extract has no cytotoxic side effects on NCM460 in the dose range of 0-40 μ g/mL, so this dose range was chosen for radiation-resistant activity.
2. Effect of duckweed extract on proliferation of NCM460 cells after irradiation:
(1) taking NCM460 cells in logarithmic growth phase, respectively with cell density of 1 × 10 4 Inoculating 100 mu L/well of 96-well plate with each mL of cell suspension;
(2) diluting the duckweed extract to be detected into different concentrations, and adding the duckweed extract according to concentration gradient within the range without toxic and side effects 4 hours before irradiation;
(3) under the condition of room temperature, the patient is irradiated to 8Gy by a Gamma acell-40 type low-dose rate research radiometer, a radiation non-administration group is used as a positive control group, and normal culture is used as a negative control group (namely an NC group). The positive drug is amifostine (amifostine) which is used for verifying the reliability of the radiation model;
(4) at 120h post irradiation, 5mg/mL MTT 20. mu.L was added to each well;
(5) after the culture, carefully sucking and discarding the supernatant, adding 100. mu.L of dimethyl sulfoxide (DMSO) into each well, shaking for 60 seconds, detecting by using a microplate reader after the reduction product of MTT is completely dissolved, and detecting the light absorption value (OD) by using a 492nm and 630nm double wavelength. Cell viability was calculated according to the following formula: cell viability (%) - [ OD average of experimental group-OD average of background group ]/[ OD average of blank group-OD average of background group ] × 100%.
Experimental data are presented as mean ± standard deviation, plotted using GraphPad software (as shown in fig. 1 and 2), and differences between groups were compared using the t-test (p <0.05, p <0.01, p < 0.001). Wherein A is the duckweed extract with the sum of the mass contents of the four caffeoyl quinic acid compounds in the duckweed extract being 5-30%, B is the refined duckweed extract obtained by further purifying the duckweed extract on the basis of A, and the sum of the mass contents of the four caffeoyl quinic acid compounds in the duckweed extract is more than 30%. The results in fig. 1 and fig. 2 show that the two duckweed extracts (a and B) have good radiation resistance (p <0.05) in the selected experimental concentration range, and the radiation resistance effect is better as the mass content of the four caffeoyl quinic acid compounds is increased.
The inventor also carried out in-vitro cell anti-radiation activity measurement on the duckweed extract (C) with the sum of the mass contents of the four caffeoylquinic acid compounds in the duckweed extract being less than 5%, and the result of figure 3 shows that the extract has no remarkable anti-radiation effect.
Biological activity tests prove that the duckweed extract can effectively prevent cell damage caused by radiation, and the further obtained refined extract product also has the same radiation damage resistance effect. The duckweed extract and the refined extract product obtained by further purifying the duckweed extract can be used as raw materials for preparing medicines, cosmetics or medical instruments with the function of resisting radiation damage.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (8)
1. Use of a duckweed extract comprising caffeoyl quinic acid compounds with a mass content of 5% or more in the preparation of a medicament having an effect of preventing radiation-induced cell damage, wherein the caffeoyl quinic acid compounds consist of 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid and 3,4, 5-O-tricaffeoylquinic acid;
the preparation method of the duckweed extract comprises the following steps:
step (1), extraction step: pulverizing dried whole herba Spirodelae, extracting with extractant to obtain primary extractive solution, concentrating, centrifuging or filtering, and collecting supernatant or filtrate;
step (2), a purification step: subjecting the supernatant or filtrate obtained in step (1) to macroporous resin adsorption, sequentially eluting with water and 5-30% lower alcohol aqueous solution to remove impurities, eluting with 30-95% lower alcohol aqueous solution, concentrating the eluate to dryness or removing lower alcohol, and drying to obtain the Spirodela delavayi extract;
in the duckweed extract obtained through the steps (1) and (2), four caffeoylquinic acid compounds: the mass content of the 3, 4-O-dicaffeoylquinic acid, the 3, 5-O-dicaffeoylquinic acid, the 4, 5-O-dicaffeoylquinic acid and the 3,4, 5-O-tricaffeoylquinic acid is more than 5 percent;
the extractant in the step (1) is at least one of water, ethanol and methanol;
the lower alcohol in the step (2) is C 1 -C 5 A lower alcohol of (2).
2. Use of the duckweed extract according to claim 1 for the preparation of a medicament having the effect of preventing radiation-induced cell damage, wherein said duckweed extract comprises 5.6% by mass of caffeoyl quinic acids, wherein said caffeoyl quinic acids consist of 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid and 3,4, 5-O-tricaffeoylquinic acid;
the preparation method of the duckweed extract comprises the following steps:
10 kilograms of dried duckweed whole herb is crushed, soaked in 50L of water for 12 hours, added with 50L of water and boiled for 2 times, each time lasts for 2 hours, extracting solutions are combined and concentrated to 50L, then the filtering solution is filtered, the filtrate passes through a 20L HP20 type macroporous resin column at the flow rate of 2BV/h, BV is the volume of a column bed of the macroporous resin, 100L of distilled water is used for leaching to remove impurities, 80L of 60% ethanol is used for elution, eluent is collected and concentrated to be dry, 64g of the duckweed extract is obtained, and the yield is 0.6% according to crude drug amount.
3. Use of a duckweed extract according to claim 1 for the preparation of a medicament for the prevention of radiation induced cell damage effects, wherein said duckweed extract comprises 13% by weight of caffeoylquinic acids, wherein said caffeoylquinic acids consist of 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid and 3,4, 5-O-tricaffeoylquinic acid;
the preparation method of the duckweed extract comprises the following steps:
crushing 10 kilograms of dried duckweed whole plant, soaking the dried duckweed whole plant in 50L of 50% ethanol for 12 hours, percolating, collecting 100L of percolate, concentrating the percolate to 20L, filtering the percolate, adding water to adjust the percolate to 50L, enabling the filtrate to pass through a 20L HZ818 type macroporous resin column at the flow rate of 2BV/h, wherein BV is the volume of a column bed of the macroporous resin, leaching the macroporous resin column with 100L of distilled water to remove impurities, eluting the macroporous resin column with 80L of 70% ethanol, collecting eluent, and concentrating the eluent to be dry to obtain 353g of the duckweed extract, wherein the yield of the duckweed extract is 3.5 percent based on crude drugs.
4. Use of the duckweed extract according to claim 1 for the preparation of a medicament having an effect of preventing radiation-induced cell damage, wherein said duckweed extract comprises 11% by mass of caffeoyl quinic acids, wherein said caffeoyl quinic acids consist of 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid and 3,4, 5-O-tricaffeoylquinic acid;
the preparation method of the duckweed extract comprises the following steps:
10 kilograms of dried duckweed whole plant is crushed, soaked for 1 hour by 200L of 70% ethanol, extracted for 2 minutes by a flash extractor, the extract is concentrated to 20L, then filtered, added with water and adjusted to 100L, the filtrate passes through a 20L NKA-2 type macroporous resin column at the flow rate of 2BV/h, BV is the volume of a column bed of the macroporous resin, 100L of distilled water is used for leaching to remove impurities, 80L of 80% ethanol is used for elution, the eluent is collected and concentrated to be dry, 404g of the duckweed extract is obtained, and the yield is 4.0 percent based on crude drugs.
5. Use of a refined duckweed extract product in the preparation of a medicament for preventing cell damage caused by radiation, wherein the refined duckweed extract product comprises caffeoyl quinic acid compounds with the mass content of more than 30%, wherein the caffeoyl quinic acid compounds at least comprise 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid and 3,4, 5-O-tricaffeoylquinic acid;
the preparation method of the refined duckweed extract comprises the following steps:
step (1), extraction step: pulverizing dried whole herba Spirodelae, extracting with extractant to obtain primary extractive solution, concentrating, centrifuging or filtering, and collecting supernatant or filtrate;
step (2), a purification step: subjecting the supernatant or filtrate obtained in step (1) to macroporous resin adsorption, sequentially eluting with water and 5-30% lower alcohol aqueous solution to remove impurities, eluting with 30-95% lower alcohol aqueous solution, concentrating the eluate to dryness or removing lower alcohol, and drying to obtain herba Spirodelae extract;
step (3), further purification step: further purifying the mass content of the four caffeoylquinic acids in the duckweed extract obtained in the step (2) to more than 30% by adopting a method comprising solvent extraction, gel column chromatography, ODS column chromatography and liquid phase preparation to obtain a refined duckweed extract product;
in the duckweed extract obtained through the steps (1) and (2), four caffeoylquinic acid compounds: the mass content of 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid and 3,4, 5-O-tricaffeoylquinic acid is more than 5%;
the extractant in the step (1) is at least one of water, ethanol and methanol;
the lower alcohol in the step (2) is C 1 -C 5 The lower alcohol of (2).
6. The use of the refined duckweed extract product of claim 5 for the preparation of a medicament for the prevention of radiation-induced cell damage, wherein said refined duckweed extract product comprises 43% by mass of caffeoylquinic acid compounds, wherein said caffeoylquinic acid compounds are comprised of 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid and 3,4, 5-O-tricaffeoylquinic acid;
the preparation method of the refined duckweed extract comprises the following steps:
crushing 10 kilograms of dried whole duckweed, soaking for 12 hours in 50L of water, adding 50L of water, decocting for 2 times, each time for 2 hours, combining extracting solutions, concentrating to 50L, then filtering, enabling filtrate to pass through a 20L HP20 type macroporous resin column at the flow rate of 2BV/h, wherein BV is the volume of a column bed of the macroporous resin, leaching with 100L of distilled water to remove impurities, eluting with 80L of 60% ethanol, collecting eluent, concentrating to be dry, and obtaining 64g of duckweed extract, wherein the yield is 0.6 percent based on crude drugs; the sum of the mass contents of the four caffeoylquinic acid compounds in the duckweed extract is 5.6%;
and further purifying the duckweed extract by using a gel column to obtain a refined duckweed extract product, wherein the sum of the mass contents of the four caffeoyl quinic acid compounds in the refined duckweed extract product is 43%.
7. The use of the refined duckweed extract product according to claim 5, wherein the refined duckweed extract product comprises 32% by mass of caffeoyl quinic acid compounds, wherein the caffeoyl quinic acid compounds consist of 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid and 3,4, 5-O-tricaffeoylquinic acid;
the preparation method of the refined duckweed extract comprises the following steps:
crushing 10 kilograms of dried duckweed whole plant, soaking the dried duckweed whole plant in 50L of 50% ethanol for 12 hours, percolating, collecting 100L of percolate, concentrating the percolate to 20L, filtering the percolate, adding water to adjust the percolate to 50L, enabling the filtrate to pass through a 20L HZ818 type macroporous resin column at the flow rate of 2BV/h, wherein BV is the volume of a column bed of the macroporous resin, leaching the macroporous resin column with 100L of distilled water to remove impurities, eluting the macroporous resin column with 80L of 70% ethanol, collecting eluent, concentrating the eluent to be dry to obtain 353g of duckweed extract, and the yield of the crude drug is 3.5 percent; the sum of the mass contents of the four caffeoylquinic acid compounds in the duckweed extract is 13%.
And (3) further purifying the duckweed extract by using an ODS column to obtain a refined duckweed extract product, wherein the sum of the mass contents of the four caffeoyl quinic acid compounds in the refined duckweed extract product is 32%.
8. The use of the refined duckweed extract product of claim 5 for the preparation of a medicament for the prevention of radiation-induced cell damage, wherein said refined duckweed extract product comprises 55% by mass of caffeoylquinic acid compounds, wherein said caffeoylquinic acid compounds are comprised of 3, 4-O-dicaffeoylquinic acid, 3, 5-O-dicaffeoylquinic acid, 4, 5-O-dicaffeoylquinic acid and 3,4, 5-O-tricaffeoylquinic acid;
the preparation method of the refined duckweed extract comprises the following steps:
crushing 10 kilograms of dried whole duckweed, soaking for 1 hour by using 200L of 70% ethanol, extracting for 2 minutes by using a flash extractor, concentrating the extracting solution to 20L, then filtering, adding water to adjust to 100L, allowing the filtrate to pass through a 20L NKA-2 type macroporous resin column at the flow rate of 2BV/h, wherein BV is the volume of a column bed of the macroporous resin, leaching with 100L of distilled water to remove impurities, eluting with 80L of 80% ethanol, collecting the eluent, concentrating to dryness to obtain 404g of duckweed extract, and the yield is 4.0 percent based on crude drugs; the sum of the mass contents of the four caffeoylquinic acid compounds in the duckweed extract is 11%;
further extracting the duckweed extract by using a solvent and purifying the extract by using a gel column chromatography to obtain a refined duckweed extract product; the sum of the mass contents of the four caffeoylquinic acid compounds in the refined duckweed extract product is 55%.
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