A method for extracting active ingredients from gynostemma pentaphyllum

Abstract

This invention provides a method for extracting active ingredients from *Erigeron breviscapus*, belonging to the field of pharmaceutical technology. Based on the differences in the physicochemical properties of the compounds, this invention first achieves preliminary separation of caffeic acid esters and scutellarin, followed by further purification and separation based on the properties of the two types of substances. Specifically, this invention uses ethanol-water solutions of different concentrations to extract caffeic acid esters stepwise, and uses macroporous and microporous resins for preliminary separation of the caffeic acid esters. Scutellarin is then preliminarily separated through acid-base treatment. Finally, high-purity scutellarin, 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, and scutellarin are obtained through reversed-phase industrial chromatography. This invention achieves the extraction and separation of multiple active ingredients from *Erigeron breviscapus*, and all solvents used in the extraction and separation process are recyclable, resulting in low cost.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to a method for extracting active ingredients from *Erigeron breviscapus*. Background Technology

[0002] *Erigeron breviscapus* (Vaniot) Hand.-Mazz., also known as *Erigeron breviscapus*, is the whole herb of *Erigeron breviscapus*, a plant in the genus *Erigeron* of the Asteraceae family. It was first recorded in *Diannan Bencao* (a traditional Chinese medicine text) and used to treat sequelae of stroke. *Erigeron breviscapus* contains various chemical components, among which the main active ingredients include scutellarin, 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, and erigosterol B. The chemical formulas of these five active ingredients are shown below:

[0003]

[0004] Erigeron breviscapine is a flavonoid glycoside and the earliest discovered active ingredient in *Erigeron breviscapine*. However, it has low solubility in organic solvents such as water, methanol, ethanol, and acetone, making its separation and purification difficult. Patent CN1053609A discloses an extraction process for esterase B, using 40-85% ethanol, followed by acid-base treatment. The crude product is then washed with 95% ethanol to remove impurities. This method represents the earliest and most basic extraction process, but it does not address the separation of other active ingredients.

[0005] 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, and scutellarin B are caffeic acid esters. Patent CN1136434A discloses the extraction of *Erigeron breviscapus* herb using 95% ethanol, followed by extraction with petroleum ether, ethyl acetate, and n-butanol. Separation of the ethyl acetate fraction yields 3,5-dicaffeoylquinic acid and 3,4-dicaffeoylquinic acid, but does not report methods for separating other caffeic acid esters or scutellarin B. Patent CN1462750A discloses extraction with ethanol or acetone-water, preliminary separation with macroporous resin, and repeated purification with Sephdex HL-20 to obtain scutellarin B, but also does not report methods for separating other caffeic acid esters or scutellarin B.

[0006] It is evident that existing extraction processes can only extract one or two active ingredients from the medicinal herb *Erigeron breviscapus*, without fully considering the comprehensive utilization of resources. Summary of the Invention

[0007] The purpose of this invention is to provide a method for extracting active ingredients from *Erigeron breviscapus*. This invention achieves the extraction and separation of *Erigeron breviscapus* extracts, including ethyl styraxate, 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, and ethyl styraxate.

[0008] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0009] This invention provides a method for extracting active ingredients from *Erigeron breviscapus*, wherein the active ingredients are ethyl styrax, 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, and styrax breviscapus extract, comprising the following steps:

[0010] The herb *Erigeron breviscapus* was extracted with a 95 wt% ethanol aqueous solution to obtain extract residue and extract A; the extract residue was then extracted with a 50 wt% ethanol aqueous solution to obtain extract B.

[0011] Extract A was loaded onto a macroporous resin column and eluted sequentially with water, 25 wt% ethanol aqueous solution, 50 wt% ethanol aqueous solution and 95 wt% ethanol aqueous solution. The eluent obtained from elution with 25 wt% ethanol aqueous solution and the eluent obtained from elution with 50 wt% ethanol aqueous solution were collected and concentrated to dryness to obtain fractions A-2 and A-3.

[0012] The A-2 component was loaded onto a microporous resin column and eluted sequentially with 10 wt% ethanol aqueous solution, 25 wt% ethanol aqueous solution and 95 wt% ethanol aqueous solution. The eluent obtained from the 25 wt% ethanol aqueous solution elution was collected and concentrated to dryness to obtain the A-2-2 component.

[0013] The A-2-2 component was purified by reverse-phase chromatography using methanol and 0.1 vol% formic acid aqueous solution in a volume ratio of 30:70 and acetonitrile and 0.1 vol% formic acid aqueous solution in a volume ratio of 15:85, respectively, to obtain fenugreek ester B.

[0014] The A-3 component was purified by reversed-phase chromatography using a mobile phase of methanol and 0.1 vol% formic acid aqueous solution at a volume ratio of 30:70 to obtain 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid, respectively.

[0015] The extract B was dissolved in water and the pH was adjusted to 1-2 with an acid reagent. After acidification, a precipitate was formed and separated to obtain the precipitate as component B-1.

[0016] The B-1 component was dissolved with an alkaline reagent and the pH was adjusted to 6-7 with an acid reagent. Then, it was purified by reversed-phase chromatography using methanol and 0.1 vol% formic acid aqueous solution at a volume ratio of 30:70 as the mobile phase to obtain scutellarin.

[0017] Preferably, the conditions for extracting Asarum lanceolatum using 95wt% ethanol aqueous solution include: three extractions, each extraction lasting 2 hours, each extraction at reflux temperature, and a ratio of 700L:100kg of the amount of 95wt% ethanol aqueous solution used to extract Asarum lanceolatum each time.

[0018] Preferably, the macroporous resin used in the macroporous resin column is DM130.

[0019] Preferably, the microporous resin used in the microporous resin column is HP20SS.

[0020] Preferably, the conditions for extracting the medicinal residue with a 50wt% ethanol aqueous solution include: extraction twice, extraction time of 2 hours each time, and reflux temperature each time; based on the amount of *Erigeron breviscapus* used, the ratio of the 50wt% ethanol aqueous solution to *Erigeron breviscapus* used in each extraction is 600L:100kg.

[0021] Preferably, the acid reagent used to adjust the pH value to 1-2 is sulfuric acid.

[0022] Preferably, the acidification treatment time is 24 to 48 hours.

[0023] Preferably, the alkaline reagent is a sodium hydroxide solution.

[0024] Preferably, the acid reagent used to adjust the pH value to 6-7 is sulfuric acid.

[0025] Preferably, the chromatography column used for reverse-phase chromatography purification of components A-2-2, A-3 and B-1 is a dynamic axial compression column; the packing material used in the dynamic axial compression column is C-18 reverse separation material.

[0026] This invention provides a method for extracting active ingredients from *Erigeron breviscapus*. Based on the differences in the physicochemical properties of the compounds, this invention first achieves preliminary separation of caffeic acid esters and scutellarin, followed by further purification and separation based on the properties of the two substances. Specifically, this invention uses ethanol-water solutions of different concentrations to extract caffeic acid esters stepwise, and uses macroporous and microporous resins for preliminary separation and purification of the caffeic acid esters. Scutellarin is then preliminarily separated through acid-base treatment. Finally, high-purity scutellarin, 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, and scutellarin are obtained through reversed-phase chromatography. This invention achieves high-purity extraction and separation of multiple active ingredients from *Erigeron breviscapus*, and all solvents used in the extraction and separation process are recyclable, resulting in low cost. The examples show that the purity of the erythritol ethyl acetate, 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid and scutellarin ethyl acetate obtained according to the method provided by the present invention is not less than 99%. Attached Figure Description

[0027] Figure 1 This is a flowchart of the extraction of active ingredients from *Erigeron breviscapus* in Example 1;

[0028] Figure 2 This is a physical image of the liquid phase preparation system used in Example 1 by Hanbang Industrial. Detailed Implementation

[0029] This invention provides a method for extracting active ingredients from *Erigeron breviscapus*, wherein the active ingredients are ethyl styrax, 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, and styrax breviscapus extract, comprising the following steps:

[0030] The herb *Erigeron breviscapus* was extracted with a 95 wt% ethanol aqueous solution to obtain extract residue and extract A; the extract residue was then extracted with a 50 wt% ethanol aqueous solution to obtain extract B.

[0031] Extract A was loaded onto a macroporous resin column and eluted sequentially with water, 25 wt% ethanol aqueous solution, 50 wt% ethanol aqueous solution and 95 wt% ethanol aqueous solution. The eluent obtained from elution with 25 wt% ethanol aqueous solution and the eluent obtained from elution with 50 wt% ethanol aqueous solution were collected and concentrated to dryness to obtain fractions A-2 and A-3.

[0032] The A-2 component was loaded onto a microporous resin column and eluted sequentially with 10 wt% ethanol aqueous solution, 25 wt% ethanol aqueous solution and 95 wt% ethanol aqueous solution. The eluent obtained from the 25 wt% ethanol aqueous solution elution was collected and concentrated to dryness to obtain the A-2-2 component.

[0033] The A-2-2 component was purified by reverse-phase chromatography using methanol and 0.1 vol% formic acid aqueous solution in a volume ratio of 30:70 and acetonitrile and 0.1 vol% formic acid aqueous solution in a volume ratio of 15:85, respectively, to obtain fenugreek ester B.

[0034] The A-3 component was purified by reversed-phase chromatography using a mobile phase of methanol and 0.1 vol% formic acid aqueous solution at a volume ratio of 30:70 to obtain 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid, respectively.

[0035] The extract B was dissolved in water and the pH was adjusted to 1-2 with an acid reagent. After acidification, a precipitate was formed and separated to obtain the precipitate as component B-1.

[0036] The B-1 component was dissolved with an alkaline reagent and the pH was adjusted to 6-7 with an acid reagent. Then, it was purified by reversed-phase chromatography using methanol and 0.1 vol% formic acid aqueous solution at a volume ratio of 30:70 as the mobile phase to obtain scutellarin.

[0037] In this invention, unless otherwise specified, all raw materials used are commercially available products known to those skilled in the art or prepared using methods known to those skilled in the art.

[0038] This invention uses a 95wt% ethanol-water solution to extract *Erigeron breviscapus*, obtaining extracted residue and extract A. In this invention, the *Erigeron breviscapus* is preferably pulverized before extraction. This invention does not have specific limitations on the pulverization method or the particle size of the pulverized *Erigeron breviscapus*; methods and particle sizes well-known to those skilled in the art can be used. In this invention, the preferred conditions for extraction with a 95wt% ethanol-water solution include: three extractions, each extraction lasting 2 hours, each extraction at reflux temperature, and a ratio of 95wt% ethanol-water solution to *Erigeron breviscapus* of 700L:100kg per extraction. In this invention, solid-liquid separation is preferably performed after each extraction, preferably by filtration. This invention preferably combines the liquid materials obtained from each solid-liquid separation and concentrates them, recovering the solvent to obtain extract A. This invention does not have specific limitations on the concentration method; concentration methods well-known to those skilled in the art can be used.

[0039] After obtaining extract A, the present invention loads extract A onto a macroporous resin column and elutes it sequentially with water, 25 wt% ethanol aqueous solution, 50 wt% ethanol aqueous solution, and 95 wt% ethanol aqueous solution. The eluents obtained from elution with 25 wt% ethanol aqueous solution and 50 wt% ethanol aqueous solution are collected and concentrated to dryness to obtain components A-2 and A-3. In the present invention, the macroporous resin used in the macroporous resin column is preferably DM130; the preferred ratio of extract A to macroporous resin is 17.2 kg: 319 L. Preferably, extract A is dissolved in 95 wt% ethanol aqueous solution, mixed with a portion of macroporous resin, vacuum dried, and then loaded onto a macroporous resin column containing the remaining macroporous resin, and eluted with the above-mentioned eluent; the preferred volume ratio of the portion of macroporous resin to the remaining macroporous resin is 69: 250. In this invention, the eluent obtained by elution with water and 95wt% ethanol aqueous solution does not contain the main active ingredients. The eluent obtained by elution with water is discarded directly, while the eluent obtained by elution with 95wt% ethanol aqueous solution can be discarded after solvent recovery.

[0040] After obtaining component A-2, this invention loads component A-2 onto a microporous resin column and elutes it sequentially with 10 wt% ethanol aqueous solution, 25 wt% ethanol aqueous solution, and 95 wt% ethanol aqueous solution. The eluent obtained from elution with 25 wt% ethanol aqueous solution is collected, concentrated to dryness, and then component A-2-2 is obtained. In this invention, the microporous resin used in the microporous resin column is preferably HP20SS; the preferred ratio of component A-2 to microporous resin is 2.95 kg: 11.8 L. Preferably, component A-2 is dissolved in 95 wt% ethanol aqueous solution, mixed with microporous resin, vacuum dried, and then loaded onto a microporous resin column for elution with the aforementioned eluent. In this invention, the eluents obtained from elution with 10 wt% ethanol aqueous solution and 95 wt% ethanol aqueous solution do not contain the main active ingredient and can be discarded after solvent recovery.

[0041] After obtaining the extracted residue, the present invention uses a 50wt% ethanol aqueous solution to extract the residue, yielding extract B. In the present invention, the preferred conditions for extracting the residue using a 50wt% ethanol aqueous solution include: extraction twice, each extraction lasting 2 hours, and extraction at reflux temperature; based on the amount of *Erigeron breviscapus* used, the ratio of the 50wt% ethanol aqueous solution to *Erigeron breviscapus* used in each extraction is 600L:100kg. In the present invention, solid-liquid separation is preferably performed after each extraction, preferably by filtration. The present invention preferably combines the liquid materials obtained from each solid-liquid separation and concentrates them, recovering the solvent to obtain extract B. The present invention does not specifically limit the concentration method; any concentration method well known to those skilled in the art can be used.

[0042] After obtaining extract B, the present invention dissolves extract B in water and adjusts the pH value to 1-2 using an acid reagent. The precipitate is then precipitated by acidification, and the precipitate is separated to obtain component B-1. In the present invention, the acid reagent used to adjust the pH value to 1-2 is preferably sulfuric acid; the acidification time is preferably 24-48 hours, and the temperature is preferably room temperature.

[0043] After obtaining components A-2-2, A-3, and B-1, the present invention performs reverse-phase chromatography purification on components A-2-2, A-3, and B-1 respectively to obtain high-purity target active ingredients, which will be described in detail below.

[0044] After obtaining component A-2-2, this invention sequentially uses methanol and 0.1 vol% formic acid aqueous solution (volume ratio 30:70) as the mobile phase and acetonitrile and 0.1 vol% formic acid aqueous solution (volume ratio 15:85) as the mobile phase to purify component A-2-2 by reversed-phase chromatography to obtain acetonitrile ethyl acetate. Specifically, this invention first uses methanol and 0.1% formic acid aqueous solution (volume ratio 30:70) as the mobile phase for elution, and then uses acetonitrile and 0.1% formic acid aqueous solution (volume ratio 15:85) as the mobile phase for elution. In this invention, the chromatography column used for the reversed-phase chromatography purification is preferably a dynamic axial compression column, and the flow rate is preferably 300-600 mL / min, more preferably 450 mL / min; the packing material used in the dynamic axial compression column is preferably C-18 reverse separation material. In this invention, when performing reversed-phase chromatography purification, it is preferable to weigh 100g of sample each time. Specifically, the weighed sample is dissolved in 30wt% methanol aqueous solution and then purified by reversed-phase chromatography to obtain fenugreel ethyl.

[0045] After obtaining component A-3, this invention uses a mobile phase of methanol and 0.1 vol% formic acid aqueous solution at a volume ratio of 30:70 to purify component A-3 by reversed-phase chromatography, yielding 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid, respectively. In this invention, the chromatographic column used for the reversed-phase chromatography purification is preferably a dynamic axial compression column; the packing material used in the dynamic axial compression column is preferably C-18 reverse separation material. In this invention, it is preferable to weigh 100 g of sample each time for reversed-phase chromatography purification, specifically by dissolving the weighed sample in a 30 wt% methanol aqueous solution and then performing reversed-phase chromatography purification to obtain 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid, respectively.

[0046] After obtaining component B-1, the present invention specifically involves dissolving component B-1 with an alkaline reagent, adjusting the pH to 6-7 with an acidic reagent, and then performing reversed-phase chromatography purification using a mobile phase of methanol and 0.1 vol% formic acid aqueous solution at a volume ratio of 30:70 to obtain scutellarin. In this invention, the alkaline reagent is preferably a sodium hydroxide solution, preferably with a concentration of 10 wt%; the acidic reagent used to adjust the pH to 6-7 is preferably sulfuric acid, preferably with a concentration of 30 wt%. In this invention, the chromatographic column used for the reversed-phase chromatography purification is preferably a dynamic axial compression column; the packing material used in the dynamic axial compression column is preferably C-18 reverse separation material. In this invention, it is preferable to weigh 100 g of sample each time for reversed-phase chromatography purification. Specifically, the weighed sample is dissolved in sodium hydroxide solution, the pH is adjusted to 6-7 with sulfuric acid, and then reversed-phase chromatography purification is performed to obtain scutellarin.

[0047] The following will refer to embodiments of the present invention (process flow diagram as shown). Figure 1 As shown, the technical solutions of this invention are clearly and completely described. Obviously, the described embodiments are only a part of the embodiments of this invention, and not all of them. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0048] Example 1

[0049] 100 kg of dried *Erigeron breviscapus* was pulverized and extracted three times by reflux with 95 wt% ethanol aqueous solution. Each extraction used 700 L of 95 wt% ethanol aqueous solution and lasted for 2 hours. The extracts from the three extractions were combined, and the total extract was concentrated and the solvent was recovered to obtain 17.2 kg of extract A. The residue obtained after the three extractions was extracted twice by reflux with 50 wt% ethanol aqueous solution. Each extraction used 600 L of 50 wt% ethanol aqueous solution and lasted for 2 hours. The extracts from the two extractions were combined, and the total extract was concentrated and the solvent was recovered to obtain 16.6 kg of extract B. Extract B was dissolved in water and acidified with sulfuric acid to pH 2. After standing for 24 hours, a precipitate formed. The precipitate was separated and obtained as B-1 (5.1 kg, crude *Erigeron breviscapus* ethyl acetate).

[0050] Extract A (17.2 kg) was dissolved in 95 wt% ethanol aqueous solution and mixed with 69 L macroporous resin (DM130). The mixture was then vacuum dried and packed into a 250 L macroporous resin column. The dried mixture was then subjected to chromatography for preliminary separation of caffeic acid phenolic compounds. Specifically, the mixture was eluted sequentially with water, 25 wt% ethanol aqueous solution, 50 wt% ethanol aqueous solution, and 95 wt% ethanol aqueous solution. The eluents obtained from elution with 25 wt% ethanol aqueous solution and 50 wt% ethanol aqueous solution were collected (the eluent obtained from water elution did not contain the main active ingredients and was discarded directly; the eluent obtained from elution with 95 wt% ethanol aqueous solution mainly contained pigments and other impurities and could be discarded after solvent recovery). The eluents were concentrated to dryness to obtain A-2 (2.95 kg, mainly containing ethyl fenugreekate) and A-3 (2.35 kg, mainly containing 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, and 4,5-dicaffeoylquinic acid).

[0051] A-2 (2.95 kg) was dissolved in 95 wt% ethanol aqueous solution and mixed with 11.8 L of microporous resin (HP20SS). The mixture was then vacuum dried. The dried mixture was loaded onto a microporous resin column and eluted sequentially with 10 wt% ethanol aqueous solution, 25 wt% ethanol aqueous solution, and 95 wt% ethanol aqueous solution. The eluent obtained from elution with 25 wt% ethanol aqueous solution (the eluents obtained from elution with 10 wt% ethanol aqueous solution and 95 wt% ethanol aqueous solution do not contain the main active ingredient and can be discarded after solvent recovery) was collected and concentrated to obtain A-2-2 (1088 g, the main component of which is ethyl fenugreek).

[0052] The A-2-2, A-3, and B-1 were purified by reversed-phase chromatography using the Hanbang Industrial Preparative Liquid Chromatography System (as shown in Figure 2). The chromatographic column used was a Dynamic Axial Compression (DAC) column. Specifically, 10 kg of C-18 reverse separation material was packed into the DAC column, and the flow rate was controlled at 450 mL / min. The specific steps are as follows:

[0053] Each time, 100g of sample A-2-2 (1088g) was weighed and dissolved in 30wt% methanol aqueous solution. Then, reversed-phase chromatography was performed for purification. Specifically, methanol and 0.1% formic acid aqueous solution (volume ratio of methanol to formic acid aqueous solution was 30:70) were used as the mobile phase for elution. Then, acetonitrile and 0.1% formic acid aqueous solution (volume ratio of acetonitrile to formic acid aqueous solution was 15:85) were used as the mobile phase for elution. Finally, 130g of fenugreek ethyl acetate with a purity of not less than 99% was obtained.

[0054] Each time, 100g of sample A-3 (2350g) was weighed and dissolved in 30wt% methanol aqueous solution, and then purified by reversed-phase chromatography. Specifically, methanol and 0.1% formic acid aqueous solution were used as the mobile phase (volume ratio of methanol to formic acid aqueous solution was 30:70) to elute and obtain 150g of 3,5-dicaffeoylquinic acid, 140g of 3,4-dicaffeoylquinic acid and 100g of 4,5-dicaffeoylquinic acid with a purity of not less than 99%.

[0055] Each time, 100g of sample B-1 (5100g) was weighed and dissolved in 10wt% sodium hydroxide aqueous solution. Then, the pH was adjusted to 7 with 30wt% sulfuric acid to obtain an acid-base treatment solution. The acid-base treatment solution was purified by reversed-phase chromatography. Specifically, methanol and 0.1% formic acid aqueous solution were used as the mobile phase (volume ratio of methanol to formic acid aqueous solution was 30:70) for elution to obtain 250g of pure scutellaria baicalensis ethylsin with a purity of not less than 99%.

[0056] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for extracting active ingredients from *Erigeron breviscapus*, wherein the active ingredients are ethyl filtrate, 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, and ethyl filtrate, comprising the following steps: The herb *Erigeron breviscapus* was extracted with a 95 wt% ethanol aqueous solution to obtain extract residue and extract A; the extract residue was then extracted with a 50 wt% ethanol aqueous solution to obtain extract B. Extract A was loaded onto a macroporous resin column and eluted sequentially with water, 25 wt% ethanol aqueous solution, 50 wt% ethanol aqueous solution and 95 wt% ethanol aqueous solution. The eluent obtained from elution with 25 wt% ethanol aqueous solution and the eluent obtained from elution with 50 wt% ethanol aqueous solution were collected and concentrated to dryness to obtain fractions A-2 and A-3. The A-2 component was loaded onto a microporous resin column and eluted sequentially with 10 wt% ethanol aqueous solution, 25 wt% ethanol aqueous solution and 95 wt% ethanol aqueous solution. The eluent obtained from the 25 wt% ethanol aqueous solution elution was collected and concentrated to dryness to obtain the A-2-2 component. The A-2-2 component was purified by reverse-phase chromatography using methanol and 0.1 vol% formic acid aqueous solution in a volume ratio of 30:70 and acetonitrile and 0.1 vol% formic acid aqueous solution in a volume ratio of 15:85, respectively, to obtain fenugreek ester B. The A-3 component was purified by reversed-phase chromatography using a mobile phase of methanol and 0.1 vol% formic acid aqueous solution at a volume ratio of 30:70 to obtain 3,5-dicaffeoylquinic acid, 3,4-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid, respectively. The extract B was dissolved in water and the pH was adjusted to 1-2 with an acid reagent. After acidification, a precipitate was formed and separated to obtain the precipitate as component B-1. The B-1 component was dissolved with an alkaline reagent and the pH was adjusted to 6-7 with an acid reagent. Then, it was purified by reversed-phase chromatography using methanol and 0.1 vol% formic acid aqueous solution at a volume ratio of 30:70 as the mobile phase to obtain scutellarin.

2. The method according to claim 1, characterized in that, The conditions for extracting Asarum lanceolatum using 95wt% ethanol aqueous solution include: three extractions, each extraction lasting 2 hours, each extraction at reflux temperature, and a ratio of 700L:100kg of 95wt% ethanol aqueous solution to Asarum lanceolatum in each extraction.

3. The method according to claim 1 or 2, characterized in that, The macroporous resin used in the macroporous resin column is DM130.

4. The method according to claim 3, characterized in that, The microporous resin used in the microporous resin column is HP20SS.

5. The method according to claim 1, characterized in that, The conditions for extracting the medicinal residue using a 50wt% ethanol aqueous solution include: two extractions, each extraction lasting 2 hours, and each extraction at a reflux temperature; based on the amount of *Erigeron breviscapus* used, the ratio of the 50wt% ethanol aqueous solution to *Erigeron breviscapus* used in each extraction is 600L:100kg.

6. The method according to claim 1, characterized in that, The acid reagent used to adjust the pH value to 1-2 is sulfuric acid.

7. The method according to claim 1 or 6, characterized in that, The acidification treatment time is 24 to 48 hours.

8. The method according to claim 1, characterized in that, The alkaline reagent is a sodium hydroxide solution.

9. The method according to claim 1 or 8, characterized in that, The acid used to adjust the pH value to 6-7 is sulfuric acid.

10. The method according to claim 1, characterized in that, The chromatography column used for reversed-phase chromatography purification of components A-2-2, A-3 and B-1 is a dynamic axial compression column; the packing material used in the dynamic axial compression column is C-18 reversed-phase separation material.

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