A method for extracting multiple effective components from spinach seeds and its application

Abstract

The invention provides a method and application for extracting multiple effective components from Chinese jujube seeds, belonging to the field of biomedical technology. The method uses D101 macroporous adsorption resin and ethanol of different concentrations to separate and extract flavonoids, saponins and free triterpenoid components from Chinese jujube seeds extract, and conducts pharmacodynamic investigation on each separated and purified part. The flavonoids, saponins and free triterpenoid components extracted from Chinese jujube seeds all have the effects of nourishing the heart and calming the nerves, sedation and hypnosis on mice with insomnia caused by para-chlorophenylalanine, among which the sedation and hypnosis effect of Chinese jujube seeds saponin components is more significant. The method has simple process, high efficiency, high recovery rate, safe separation materials, and the macroporous resin materials used can be repeatedly used for many times, which is safe and has low production cost, and provides a good technical solution for the development of "safe, effective and controllable" Chinese jujube seeds effective part raw materials.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine and relates to a method for extracting multiple effective components from wild jujube seeds and application thereof. Background Art

[0002] Semen ziziphi spinosae (SZS) is the dried mature seed of the Rhamnaceae shrub Ziziphusjujuba Mill. Var. spinosa (Bunge) Hu ex HF Chou. It is sweet and sour in taste and neutral in nature. It has the effects of nourishing the heart and liver, calming the mind, astringing sweat and promoting body fluid. Modern pharmacological studies have shown that Semen ziziphi spinosae has the effects of anti-anxiety, anticonvulsant, sedative and hypnotic, enhancing immune function, improving cognitive function, anti-atherosclerosis, anti-inflammatory, and myocardial protection.

[0003] The main components of the seeds of the Chinese jujube are flavonoids, triterpenes, volatile oils and organic acids. Among them, the flavonoid components are a series of compounds with 2-phenylchromone as the basic mother nucleus, which generally refers to natural products with two benzene rings connected by a central three-carbon chain, that is, with a C6-C3-C6 basic mother nucleus. The flavonoid components in the seeds of the Chinese jujube mainly include spinosin, 6'''-feruloyl spinosin, etc., which have pharmacological effects such as sedation and hypnosis, anti-depression, anti-anxiety, antioxidant, enhanced learning and memory, and brain nerve protection. The triterpenoid components are a class of terpenoid compounds with a basic mother nucleus composed of 30 carbon atoms. The triterpenoid components in the seeds of the Chinese jujube are mostly bound dammarane-type tetracyclic triterpenoid saponins, such as jujube saponin A and jujube saponin B, which have sedation and hypnosis, anticonvulsant, hypolipidemic and cardiovascular system protection effects. The free triterpenoid components in the seeds of the Chinese date include betulinic acid, betulinol, theophyllic acid, and maltose acid, which are lupane-type pentacyclic triterpenoids. In addition to their anti-inflammatory, anti-tumor, antibacterial, and anti-HIV pharmacodynamic activities, they also have certain antidepressant, sedative, and hypnotic effects. The flavonoids, saponins, and free triterpenoids in the seeds of the Chinese date all have certain sedative and hypnotic effects, but they are quite different in other pharmacodynamic activities, and their structures and physicochemical properties are also quite different.

[0004] Regarding the extraction of effective ingredients such as flavonoids from Ziziphus jujuba seeds, the Chinese patent with application number 201910717516.0 discloses the use of reverse phase chromatography fillers to separate and purify the two major active ingredients of flavonoids and saponins in Ziziphus jujuba seeds extract by adjusting the proportion of organic phase in the mobile phase. The Chinese patent with application number 201510137286.2 discloses the use of HPD100 macroporous adsorption resin to separate and purify the effective parts of flavonoids and saponins. However, the above patents only separate and purify the flavonoids and saponins components in Ziziphus jujuba seeds, and do not involve free triterpenoid components, nor do they investigate the efficacy of the separated and purified components. Summary of the invention

[0005] The purpose of the present invention is to provide a method for extracting multiple effective components from spinach seeds and application thereof, so as to solve the problem that the existing extraction method fails to extract free triterpenoid components.

[0006] To achieve the above object, the present invention adopts the following technical solutions:

[0007] The present application provides a method for extracting multiple effective ingredients from spinach seeds, the method comprising:

[0008] S01: Add 5 times the amount of 70% ethanol to the Ziziphus jujuba seed powder, heat and reflux for 1-1.5 hours, filter, and concentrate under reduced pressure to obtain a concentrated solution, in which the amount of Ziziphus jujuba seed crude drug is 0.2-0.4 g / mL and the alcohol content is 20-40%.

[0009] The dried Chinese jujube kernels were crushed to form Chinese jujube kernel powder. Five times the volume of 70% ethanol was added to the Chinese jujube kernel powder, and the mixture was heated to reflux for 1-1.5 hours at 90°C. After the reflux, the solution was filtered to obtain a filtrate. The filtrate was concentrated under reduced pressure at 50°C until the amount of Chinese jujube kernel crude drug in the filtrate was 0.2-0.4 g / mL and the alcohol content was 20-40%, thereby obtaining a concentrated solution.

[0010] S02: The concentrated solution was loaded onto a macroporous adsorption resin column, and after standing for 40-60 minutes of adsorption, it was eluted in sequence with 5 BV 40% ethanol, 6 BV 60% ethanol and 8 BV 90% ethanol, and the eluates were collected respectively.

[0011] The concentrate is loaded onto a macroporous adsorption resin column, wherein the loading amount of the concentrate is 5 times the volume BV of the macroporous adsorption resin column, and the loading rate is 2 BV / h. After loading, stand for 40-60 minutes to allow the macroporous adsorption resin column to fully adsorb the concentrate. In the present application, the macroporous adsorption resin column is a D101 macroporous adsorption resin column. The D101 macroporous adsorption resin belongs to a non-polar resin, which is a spherical styrene-type non-polar copolymer. It has a generally strong adsorption capacity for organic compounds that are not polar or weakly polar, and is particularly good for the separation and purification of saponins and flavonoid components. It is mostly used in the preparation of natural active ingredients of Chinese herbal medicine. In the present application, D101 macroporous adsorption resin is used to separate flavonoid components, saponin components and free triterpenoid components in Ziziphus jujuba seeds.

[0012] After standing, 5 BV 40% ethanol, 6 BV 60% ethanol and 8 BV 90% ethanol were used to elute the D101 macroporous adsorption resin column at an elution rate of 5 BV / h, and the eluates were collected to obtain 40% ethanol eluate, 60% ethanol eluate and 90% ethanol eluate.

[0013] S03: The eluate is concentrated under reduced pressure and dried into powder to obtain flavonoid components, saponin components and free triterpenoid components.

[0014] The 40% ethanol eluate, the 60% ethanol eluate, and the 90% ethanol eluate were concentrated under reduced pressure, and dried at 70°C to form powders to obtain flavonoid components, saponin components, and free triterpenoid components.

[0015] The prepared flavonoids, saponins and free triterpenes were analyzed by UPLC-U3000 ultra-high performance liquid chromatography system to determine their composition and content. Figure 1-3 . Among them, the UPLC-U3000 ultra-high performance liquid chromatography analysis conditions are as follows: chromatographic column: Waters ACQUITY UPLC BEH C18 (100×2.1mm, 1.7μm), mobile phase: 0.1% phosphoric acid aqueous solution (A)-acetonitrile (B), elution conditions: 0-15min: 10%-15%B; 15-20min: 15%-20%B; 20-30min: 20%-22%B; 30-35min: 22%-42%B; 35-39min: 42%-49% B; 39-40min: 49%-81%B; 40-47min: 81%-90%B. Column temperature: 35°C, flow rate: 0.3 mL / min, injection volume: 5μL, wavelength 203nm.

[0016] By the attached Figure 1 It can be seen that the components obtained after the 40% ethanol eluate was concentrated and dried under reduced pressure contain spinotoxin and 6'''-feruloyl spinotoxin, indicating that its components are mainly flavonoid components. The content of the flavonoid component is calculated based on the total amount of spinotoxin and 6'''-feruloyl spinotoxin. The regression equation of spinotoxin is Y1=0.1499X1-0.2245, R1 2 =0.9996, linear range is 6.2713~200.68 µg / mL. 6'''-feruloyl spinosin regression equation Y2=0.1599X2-0.1043, R2 2 =0.9992, and the linear range was 6.4503~206.41 µg / mL.

[0017] By the attached Figure 2 It can be seen that the component obtained after the 60% ethanol eluate was concentrated and dried under reduced pressure contains jujuboside A, indicating that its main component is a saponin component. The saponin component is calculated based on the content of jujuboside A. The regression equation of jujuboside A is Y3=0.0658X3-0.0444, R3 2 =0.9997, and the linear range was 6.2784~200.91 µg / mL.

[0018] By the attached Figure 3 It can be seen that the components obtained after the 90% ethanol eluate was concentrated and dried under reduced pressure contain betulinic acid, indicating that its main component is free triterpenoid components. The free triterpenoid components are calculated based on the content of betulinic acid. The regression equation of betulinic acid is Y4=0.202X4-0.7989, R4 2 =0.9992, and the linear range was 6.5047~208.15 µg / mL.

[0019] In addition, in the present application, the flavonoid components, saponin components and free triterpenoid components extracted from the Chinese date seeds are respectively used to prepare drugs with the functions of nourishing the heart and calming the nerves, and sedative and hypnotic.

[0020] The present invention has the following beneficial effects:

[0021] (1) In this method, D101 macroporous adsorption resin and different concentrations of ethanol were used to separate and extract flavonoids, saponins and free triterpenoid components from the extract of Ziziphus jujuba seeds, and the pharmacodynamics of the separated and purified parts were investigated.

[0022] (2) The flavonoids, saponins and free triterpenoid components extracted from the seeds of the Chinese jujube all have the effects of nourishing the heart and calming the nerves, as well as sedative and hypnotic effects on PCPA-induced insomnia mice. Among them, the sedative and hypnotic effects of the saponins components of the seeds of the Chinese jujube were more significant.

[0023] (3) This method has a simple process, high efficiency, high recovery rate, safe separation materials, and the macroporous resin material used can be reused many times, which is safe and has low production cost. It provides a good technical solution for the development of "safe, effective, and controllable" raw materials for the effective parts of Ziziphus jujuba seeds. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 is the chromatogram of flavonoid components;

[0025] Figure 2 is the chromatogram of saponin components;

[0026] Figure 3 is the chromatogram of free triterpenoid components;

[0027] Figure 4 This is the effect of various extraction parts of Semen Ziziphi Spinosae on the weight gain of insomnia mice before and after modeling;

[0028] Figure 5 This is the effect of various extraction parts of Ziziphus jujuba seeds on the sleep latency of insomnia mice;

[0029] Figure 6 This is a graph showing the effects of various extraction parts of Ziziphus jujuba seeds on the sleep duration of insomnia mice. DETAILED DESCRIPTION

[0030] The technical solution of the present invention is further explained and illustrated by means of specific embodiments below.

[0031] Example 1

[0032] The present invention provides a method for extracting multiple active ingredients from spinach seeds, the method comprising:

[0033] S101: Crush the dried Chinese jujube kernel to form Chinese jujube kernel powder. Add 300 ml of 70% ethanol to 60 g of Chinese jujube kernel powder, and heat to reflux at 90°C for 1 hour. After the reflux is completed, filter the solution to obtain a filtrate. Concentrate the filtrate under reduced pressure at 50°C until the amount of Chinese jujube kernel crude drug in the filtrate is 0.3 g / mL and the alcohol content is 40%, thereby obtaining a concentrated solution.

[0034] S102: Load the concentrate onto a D101 macroporous adsorption resin column, wherein the amount of concentrate loaded is 5 times the volume BV of the macroporous adsorption resin column, and the loading rate is 2 BV / h. After loading, let it stand for 50 minutes to allow the macroporous adsorption resin column to fully adsorb the concentrate. After standing, 5 BV 40% ethanol, 6 BV 60% ethanol and 8 BV 90% ethanol are used to elute the D101 macroporous adsorption resin column at an elution rate of 5 BV / h, and the eluents are collected to obtain 40% ethanol eluent, 60% ethanol eluent and 90% ethanol eluent.

[0035] S103: The 40% ethanol eluate, the 60% ethanol eluate, and the 90% ethanol eluate were concentrated under reduced pressure and dried at 70° C. to form powders to obtain flavonoid components, saponin components, and free triterpenoid components.

[0036] Quantitative analysis by UPLC-U3000 ultra-high performance liquid chromatography showed that the content of flavonoids was 28.13 mg / g, the content of saponins was 25.56 mg / g, and the content of free triterpenes was 277.99 mg / g.

[0037] Example 2

[0038] The present invention provides a method for extracting multiple active ingredients from spinach seeds, the method comprising:

[0039] S201: Crush the dried Chinese jujube kernel to form Chinese jujube kernel powder. Add 300 ml of 70% ethanol to 60 g of Chinese jujube kernel powder, and heat to reflux for 1.5 h at 90°C. After the reflux is completed, filter the solution to obtain a filtrate. Concentrate the filtrate under reduced pressure at 50°C until the amount of Chinese jujube kernel crude drug in the filtrate is 0.3 g / mL and the alcohol content is 30%, thereby obtaining a concentrated solution.

[0040] S202: Load the concentrated solution onto a D101 macroporous adsorption resin column, wherein the loading amount of the concentrated solution is 5 times the volume BV of the macroporous adsorption resin column, and the loading rate is 2 BV / h. After loading, let it stand for 50 minutes to allow the macroporous adsorption resin column to fully adsorb the concentrated solution. After standing, 5 BV 40% ethanol, 6 BV 60% ethanol and 8 BV 90% ethanol are used to elute the D101 macroporous adsorption resin column at an elution rate of 5 BV / h, and the eluents are collected to obtain 40% ethanol eluent, 60% ethanol eluent and 90% ethanol eluent.

[0041] S203: The 40% ethanol eluate, the 60% ethanol eluate, and the 90% ethanol eluate were concentrated under reduced pressure and dried at 70° C. to form powders to obtain flavonoid components, saponin components, and free triterpenoid components.

[0042] Quantitative analysis by UPLC-U3000 ultra-high performance liquid chromatography showed that the content of flavonoids was 28.01 mg / g, the content of saponins was 26.54 mg / g, and the content of free triterpenes was 279.16 mg / g.

[0043] Example 3

[0044] The present invention provides a method for extracting multiple active ingredients from spinach seeds, the method comprising:

[0045] S301: Grind the dried Chinese jujube kernel to form Chinese jujube kernel powder. Add 3000 ml 70% ethanol to 600 g Chinese jujube kernel powder, and heat to reflux for 1 hour at 90°C. After the reflux is completed, filter the solution to obtain a filtrate. Concentrate the filtrate under reduced pressure at 50°C until the amount of Chinese jujube kernel crude drug in the filtrate is 0.3 g / mL and the alcohol content is 40%, thereby obtaining a concentrated solution.

[0046] S302: Load the concentrated solution onto a D101 macroporous adsorption resin column, wherein the loading amount of the concentrated solution is 5 times the volume BV of the macroporous adsorption resin column, and the loading rate is 2 BV / h. After loading, let it stand for 60 minutes to allow the macroporous adsorption resin column to fully adsorb the concentrated solution. After standing, 5 BV 40% ethanol, 6 BV 60% ethanol and 8 BV 90% ethanol are used to elute the D101 macroporous adsorption resin column at an elution rate of 5 BV / h, and the eluents are collected to obtain 40% ethanol eluent, 60% ethanol eluent and 90% ethanol eluent.

[0047] S303: The 40% ethanol eluate, the 60% ethanol eluate, and the 90% ethanol eluate were concentrated under reduced pressure and dried at 70° C. to form powders to obtain flavonoid components, saponin components, and free triterpenoid components.

[0048] Quantitative analysis by UPLC-U3000 ultra-performance liquid chromatography showed that the content of flavonoids was 27.36 mg / g, the content of saponins was 26.03 mg / g, and the content of free triterpenes was 278.66 mg / g.

[0049] Example 4

[0050] The present invention provides a method for extracting multiple active ingredients from spinach seeds, the method comprising:

[0051] S401: Crush the dried Chinese jujube kernel to form Chinese jujube kernel powder. Add 3000 ml 70% ethanol to 600 g Chinese jujube kernel powder, and heat to reflux for 1.5 h at 90°C. After reflux, filter the solution to obtain a filtrate. Concentrate the filtrate under reduced pressure at 50°C until the amount of Chinese jujube kernel crude drug in the filtrate is 0.4 g / mL and the alcohol content is 30%, thereby obtaining a concentrated solution.

[0052] S402: Load the concentrated solution onto a D101 macroporous adsorption resin column, wherein the loading amount of the concentrated solution is 5 times the volume BV of the macroporous adsorption resin column, and the loading rate is 2 BV / h. After loading, let it stand for 50 minutes to allow the macroporous adsorption resin column to fully adsorb the concentrated solution. After standing, 5 BV 40% ethanol, 6 BV 60% ethanol and 8 BV 90% ethanol are used to elute the D101 macroporous adsorption resin column at an elution rate of 5 BV / h, and the eluents are collected to obtain 40% ethanol eluent, 60% ethanol eluent and 90% ethanol eluent.

[0053] S403: The 40% ethanol eluate, the 60% ethanol eluate, and the 90% ethanol eluate were concentrated under reduced pressure and dried at 70° C. to form powders to obtain flavonoid components, saponin components, and free triterpenoid components.

[0054] Quantitative analysis by UPLC-U3000 ultra-performance liquid chromatography showed that the content of flavonoids was 27.20 mg / g, the content of saponins was 25.89 mg / g, and the content of free triterpenes was 278.30 mg / g.

[0055] In order to verify that the flavonoid components, saponin components and free triterpenoid components extracted by the method provided in this application have the functions of nourishing the heart and calming the mind, calming and hypnotic, the flavonoid components, saponin components and free triterpenoid components were used in the examples of this application to conduct experiments on PCPA (parachlorophenylalanine)-induced insomnia mice. The specific experimental process and test results are as follows:

[0056] 1. Animal grouping and modeling

[0057] After 7 days of normal feeding and adaptation to the environment, the mice were randomly divided into normal group, model group, estazolam group, flavonoid group, saponin group, and free triterpenoid group according to body weight, with 10 mice in each group. At a fixed time every day, the model group and each drug-treated group were intraperitoneally injected with 400 mg / kg PCPA suspension for 3 consecutive days, and the normal group was intraperitoneally injected with normal saline for 3 consecutive days. After the model was established, the status of each group was observed. Except for the normal group, the mice in the other groups showed day and night disorders, mutual aggression, decreased appetite, etc., indicating that the insomnia model was successfully established.

[0058] 2. Animal medication

[0059] After successful modeling, each group was given medication. The normal group and the model group were gavaged with normal saline for 7 consecutive days; the estazolam group was gavaged with estazolam for 7 consecutive days, and the gavage amount for each mouse was 0.39 mg / kg based on body weight; the flavonoid group was gavaged with flavonoid components for 7 consecutive days, and the gavage amount for each mouse was 0.309 mg / g based on body weight, the saponin group was gavaged with saponin components for 7 consecutive days, and the gavage amount for each mouse was 0.391 mg / g based on body weight, and the free triterpenoid group was gavaged with free triterpenoid components for 7 consecutive days, and the gavage amount for each mouse was 0.1619 mg / g based on body weight. The dosage of the estazolam group was obtained by converting the dosage of animals and humans. The dosage of the flavonoid group, saponin group, and free triterpenoid group was obtained by converting the dosage of human jujube seeds and the dosage of animals and humans specified in the 2020 edition of the Pharmacopoeia of the People's Republic of China (Part I).

[0060] 3. Behavioral status and weight change results

[0061] After modeling, the status of mice in each group showed obvious differences. Among them, the mice in the normal group were in good spirits, had obvious circadian rhythms, normal appetite, and smooth fur; the mice in the model group had circadian disorders, irritability, mutual aggression, decreased appetite, and hair that was easy to fall off and dull; after oral administration of drugs in each group, the status of the mice gradually returned to normal. The changes in body weight of mice in each group before and after modeling are shown in Table 1. Figure 4 .

[0062] Table 1: Effects of purified Semen Ziziphi Spinosae on body weight of insomnia mice (n=10)

[0063]

[0064] Note: Compared with the normal group, # P<0.05; compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0065] From Table 1, Figure 4 It can be seen that the weight of mice in the normal group continued to increase steadily. Compared with the normal group, the weight gain of the model group before and after modeling was not obvious (P<0.05). Compared with the model group, the weight gain of mice in the estazolam group, flavonoid group, saponin group and free triterpenoid group was extremely obvious (P<0.001), which shows that the flavonoid components, saponin components and free triterpenoid components extracted from the spiny jujube seeds in the examples of the present application can restore the state of PCPA-induced insomnia mice and increase their weight.

[0066] 4. Pentobarbital sodium synergistic sleep experiment

[0067] One day before the last administration, mice in each group were intraperitoneally injected with 50 mg / kg sodium pentobarbital, and the sleep latency, sleep duration, and time of completion of intraperitoneal injection of each mouse were immediately recorded to observe the status of the mice. The sleep latency refers to the time from the injection of sodium pentobarbital to the disappearance of the righting reflex, and the sleep duration refers to the time from the disappearance of the righting reflex to the recovery.

[0068] Place the mouse on a soft and flat surface with its abdomen facing upward. If the mouse keeps its abdomen facing upward within 60 seconds, it is considered that the mouse's righting reflex has disappeared. Record the time point and continue to observe the mouse's state. When the mouse flips over more than three times within 30 seconds, it is considered that the righting reflex has been restored and the mouse ends its sleep. The time period from the completion of the intraperitoneal injection to the disappearance of the righting reflex is recorded as the sleep latency; the time from the disappearance of the righting reflex to the recovery of the righting reflex is the sleep duration. In addition, mice whose sleep duration is greater than 10 minutes are considered to have successfully fallen asleep. After the test is completed, the sleep latency and sleep duration of each group of mice are calculated and analyzed, and the results are shown in Table 2 and Appendix. Figure 5 , 6 .

[0069] Table 2: Effects of purified extracts from Chinese date seeds on sleep latency and sleep duration in insomnia mice (n=6)

[0070]

[0071] Note: Compared with the normal group, ## P<0.01, ### P<0.001; compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0072] From Table 2, Appendix Figure 5 , 6 It can be seen that compared with the normal group, the sleep latency in the model group was significantly prolonged (P<0.01), and the sleep duration was extremely significantly shortened (P<0.001), indicating that intraperitoneal injection of PCPA can cause insomnia in mice.

[0073] Compared with the model group, the sleep latency of mice in the estazolam group and saponin group was significantly shortened (P<0.01), and the sleep duration was extremely significantly increased (P<0.001). The sleep latency of mice in the flavonoid group and free triterpenoid group was significantly shortened (P<0.05), and the sleep duration was significantly increased. Among them, the increase time in the flavonoid group (P<0.01) was longer than that in the free triterpenoid group (P<0.05).

[0074] Compared with the estazolam group, there was no significant difference in the sleep latency and sleep duration of mice in the saponin group (P>0.05), and there was a certain gap between the flavonoid group and the free triterpenoid group and the estazolam group (P<0.05). This shows that the purified parts of Semen Ziziphi Spinosae have an improvement effect on the sleep of PCPA insomnia mice, but the improvement is different. Among them, the saponin component has an effect on improving insomnia in mice that is comparable to the drug estazolam, and is better than the flavonoid component and the free triterpenoid component.

[0075] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. The use of free triterpenoid components extracted from Ziziphus jujuba seeds in the preparation of drugs with heart-nourishing, tranquilizing, sedative and hypnotic functions, wherein the free triterpenoid components are extracted according to the following method: Add 5 times the volume of 70% ethanol to the Ziziphus jujuba seed powder, heat and reflux for 1-1.5 hours, filter, and concentrate under reduced pressure to obtain a concentrated solution, wherein the amount of Ziziphus jujuba seed crude drug in the concentrated solution is 0.2-0.4 g / mL and the alcohol content is 20-40%; The concentrated solution was loaded onto a D101 macroporous adsorption resin column, and after standing for 40-60 minutes of adsorption, 5 BV of 40% ethanol, 6 BV of 60% ethanol, and 8 BV of 90% ethanol were used for elution in sequence, and the eluates were collected respectively; The eluate is concentrated under reduced pressure and dried into powder to obtain flavonoid components, saponin components and free triterpenoid components, wherein: The free triterpenoid fraction was obtained by eluting with 8 BV 90% ethanol.

2. The use according to claim 1, characterized in that: The heating reflux temperature was 90°C.

3. The use according to claim 1, characterized in that: The loading amount of the concentrated solution is 5 times the volume BV of the macroporous adsorption resin column.

4. The use according to claim 1, characterized in that: The loading rate of the concentrate was 2 BV / h.

5. The use according to claim 1, characterized in that: The elution rate was 5 BV / h.

6. The use according to claim 1, characterized in that: The mixture was concentrated under reduced pressure at 50°C to obtain a concentrate.

7. The use according to claim 1, characterized in that: The drying temperature is 70°C.

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