A method for extracting beta-amyrin

Abstract

The application discloses a kind of β-amyrin extraction methods, including wood pod red bean, the leaf of wood pod red bean β-amyrin content is 0.18~0.31%, extract new method, including the following steps: step one, dry screening;Step two, extraction mixing;Step three, concentrated extract;Step four, impurity purification;Step five, crystallization precipitation, select wood pod red bean leaf as preparation raw material, obtain simple, reduce raw material cost, prepare β-amyrin extract by multiple extraction mixing in preparation process, fully extract β-amyrin in raw material, reduce the loss of β-amyrin in extraction process, improve yield;Different solvent ratio petroleum ether-acetic ether is used in impurity purification and is eluted twice respectively, remove impurities, improve the purity after preparation, extraction method is simple and easy to operate, reduce production difficulty, save production cost.

Description

Technical Field

[0001] This invention relates to the field of β-amyrin extraction technology, specifically to a method for extracting β-amyrin. Background Technology

[0002] β-Armyrin is a white crystalline powder with the molecular formula C2. 30 H 50 O, with a molecular weight of 426, is a pentacyclic triterpenoid compound with anti-inflammatory, antitumor, antiviral, hypoglycemic, and hypolipidemic medicinal effects. It can be used as an additive in daily chemical products and is also a synthetic intermediate for glycyrrhizic acid, a liver drug. There is a large market demand. However, β-amyrin has a complex chemical structure and multiple stereostructures. The method of preparing β-amyrin from plants such as Gentiana macrophylla, Citrus reticulata peel, Eriocaulon buergerianum, and Ipomoea quamoclit through solvent extraction, extraction, and repeated treatment with silica gel and gel column chromatography is complicated, costly, and has a very low final yield, making industrialization difficult. Therefore, it is essential to find a plant rich in β-amyrin and design a new extraction method. Summary of the Invention

[0003] The purpose of this invention is to provide a method for extracting β-amyrin to solve the problems mentioned in the background art.

[0004] To achieve the above objectives, the present invention provides the following technical solution: a plant rich in β-amyrin, comprising red bean, wherein the β-amyrin content in the leaves of the red bean plant is 0.18% to 0.31%.

[0005] A method for extracting β-amyrin includes the following steps: Step 1, drying and sieving; Step 2, extraction and mixing; Step 3, concentrating the extract; Step 4, removing impurities and purifying; Step 5, crystallization. In step one above, dried red bean leaves are taken, dried in an oven, then pulverized using a pulverizer, and then sieved through a sieve to obtain red bean leaf powder. In step two above, the leaf powder of red bean is mixed with ethyl acetate solution and poured into a heating furnace. The mixture is then heated to boiling for a period of time. After filtration, the first extract is obtained. The filter residue is then mixed with ethyl acetate solution and added to the heating furnace. The mixture is heated to boiling for a period of time and then filtered again to obtain the second extract. Finally, the first extract and the second extract are mixed to obtain the β-amyrin extract. In step three above, the β-amyrin extract obtained in step two is placed in a concentration tank and concentrated under reduced pressure to obtain an ethyl acetate extract paste. In step four above, the ethyl acetate extract obtained in step three is eluted for the first time with petroleum ether-ethyl acetate to remove impurities, and then eluted a second time with petroleum ether-ethyl acetate. The eluent is collected and placed in a concentration tank for vacuum concentration to obtain a high-concentration β-amyrin extract. In step five above, ethyl acetate-ethanol mixed solvent is added to the high-concentration β-amyrin extract obtained in step four, then the mixture is heated and allowed to stand. After standing, it is filtered to obtain β-amyrin.

[0006] Preferably, in step one, the drying temperature is 75-85℃, the drying time is 10-15 minutes, and a 20-mesh sieve is used for screening.

[0007] Preferably, in step two, the mass of the powdered leaves of *Adenium obesum* is 950–1050 g, the volume of the ethyl acetate solution mixed with the powdered leaves is 9500–10500 ml, and the boiling time is 110–130 min.

[0008] Preferably, in step two, the volume of the ethyl acetate solution mixed with the filter residue is 9500–10500 ml, and the boiling time is 110–130 min.

[0009] Preferably, in step three, the pressure in the concentration tank is 93.32–98.64 kPa, the temperature is 38–42°C, and the concentration time under reduced pressure is 110–130 min.

[0010] Preferably, in step four, the elution solvent ratio of the petroleum ether-ethyl acetate system in the initial elution is 50:1, and the elution volume is 4 to 6 column volumes.

[0011] Preferably, in step four, the ratio of the petroleum ether-ethyl acetate system as the eluting solvent is 30:1, the elution volume is 4 to 6 column volumes, the pressure in the concentration tank is 93.32 to 98.64 kPa, the temperature is 38 to 42°C, and the concentration time under reduced pressure is 110 to 130 min.

[0012] Preferably, in step five, the volume ratio of ethyl acetate to ethanol in the ethyl acetate-ethanol mixed solvent is 3.5–4.8:1, the temperature is raised to 55–60°C, and the standing time is 120–180 min.

[0013] Compared with the prior art, the beneficial effects of the present invention are as follows: This method for extracting β-amyrin uses leaves of *Adenanthera philoxeroides* as the raw material, which is easy to obtain and reduces the cost of raw materials. The preparation method involves multiple extractions and mixing to prepare a β-amyrin extract, which fully extracts β-amyrin from the raw material and improves the yield of β-amyrin. In the purification process, petroleum ether-ethyl acetate with different solvent ratios is used for two elutions to remove impurities from the material and improve the purity of the prepared β-amyrin. There is no need to use gel chromatography or liquid chromatography for separation, the preparation method is simple, the final yield is increased, and the production difficulty is reduced. Attached Figure Description

[0014] Figure 1 This is a flowchart of the extraction method of the present invention. Detailed Implementation

[0015] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0016] Please see Figure 1 One embodiment provided by the present invention: Example 1:

[0017] A plant rich in β-amyrin, including *Acer sarcodactylis*, wherein the leaves of *Acer sarcodactylis* contain 0.18% to 0.31% β-amyrin.

[0018] A method for extracting β-amyrin includes the following steps: Step 1, drying and sieving; Step 2, extraction and mixing; Step 3, concentrating the extract; Step 4, removing impurities and purifying; Step 5, crystallization. In step one above, 1000g of dried red bean leaves are taken, baked in an oven at 80℃ for 15 minutes, then crushed using a pulverizer, and then sieved through a 20-mesh sieve to obtain red bean leaf powder. In step two above, the leaf powder of red bean is mixed with 10,000 ml of ethyl acetate solution and poured into a heating furnace. The mixture is then heated to boiling for 120 minutes. After filtration, the first extract is obtained. The filter residue is then mixed with 10,000 ml of ethyl acetate solution and added to the heating furnace. The mixture is heated to boiling for 120 minutes and then filtered again to obtain the second extract. The first and second extracts are then mixed to obtain the β-amyrin extract. In step three above, the β-amyrin extract obtained in step two is placed in a concentration tank, the pressure is adjusted to 95 kPa and the temperature is 40℃, and the concentration is carried out under reduced pressure for 120 min. After concentration, an ethyl acetate extract is obtained. In step four above, the ethyl acetate extract obtained in step three is eluted for 5 column volumes with a petroleum ether-ethyl acetate eluent ratio of 50:1 as the first elution to remove impurities. Then, it is eluted again with a petroleum ether-ethyl acetate eluent ratio of 30:1 for 5 column volumes as the second elution. The eluent is collected and placed in a concentration tank. The pressure is adjusted to 96 kPa and the temperature is 40°C. The mixture is concentrated under reduced pressure for 115 min to obtain a high-concentration β-amyrin extract. In step five above, the volume ratio of ethyl acetate-ethanol mixed solvent added to the high-concentration β-amyrin extract obtained in step four is 4.5:1. Then the temperature is raised to 58°C, and the mixture is allowed to stand for 150 minutes. After standing, the mixture is filtered to obtain β-amyrin. Example 2:

[0019] A plant rich in β-amyrin, including *Acer sarcodactylis*, wherein the leaves of *Acer sarcodactylis* contain 0.18% to 0.31% β-amyrin.

[0020] A method for extracting β-amyrin includes the following steps: Step 1, drying and sieving; Step 2, extraction and mixing; Step 3, concentrating the extract; Step 4, removing impurities and purifying; Step 5, crystallization. In step one above, 1000g of dried red bean leaves are taken, baked in an oven at 82℃ for 12 minutes, then crushed using a pulverizer, and then sieved through a 20-mesh sieve to obtain red bean leaf powder. In step two above, the leaf powder of red bean is mixed with 10,000 ml of ethyl acetate solution and poured into a heating furnace. The mixture is then heated to boiling for 125 min. After filtration, the first extract is obtained. The filter residue is then mixed with 10,000 ml of ethyl acetate solution and added to the heating furnace. The mixture is heated to boiling for 115 min. After filtration, the second extract is obtained. The first and second extracts are then mixed to obtain the β-amyrin extract. In step three above, the β-amyrin extract obtained in step two is placed in a concentration tank, the pressure is adjusted to 95.56 kPa and the temperature is 40℃, and the concentration is carried out under reduced pressure for 120 min. After concentration, an ethyl acetate extract is obtained. In step four above, the ethyl acetate extract obtained in step three is eluted for 6 column volumes with a petroleum ether-ethyl acetate eluent ratio of 50:1 as the first elution to remove impurities. Then, it is eluted again with a petroleum ether-ethyl acetate eluent ratio of 30:1 for 6 column volumes as the second elution. The eluent is collected and placed in a concentration tank. The pressure is adjusted to 98.21 kPa and the temperature is 40°C. The mixture is concentrated under reduced pressure for 120 min to obtain a high-concentration β-amyrin extract. In step five above, the volume ratio of ethyl acetate-ethanol mixed solvent added to the high-concentration β-amyrin extract obtained in step four is 4.5:1. Then the temperature is raised to 58°C, and the mixture is allowed to stand for 150 minutes. After standing, the mixture is filtered to obtain β-amyrin. Example 3:

[0021] A plant rich in β-amyrin, including *Acer sarcodactylis*, wherein the leaves of *Acer sarcodactylis* contain 0.18% to 0.31% β-amyrin.

[0022] A method for extracting β-amyrin includes the following steps: Step 1, drying and sieving; Step 2, extraction and mixing; Step 3, concentrating the extract; Step 4, removing impurities and purifying; Step 5, crystallization. In step one above, 1000g of dried red bean leaves are taken, baked in an oven at 83℃ for 12 minutes, then crushed using a pulverizer, and then sieved through a 20-mesh sieve to obtain red bean leaf powder. In step two above, the leaf powder of red bean is mixed with 10,000 ml of ethyl acetate solution and poured into a heating furnace. The mixture is then heated to boiling for 120 minutes. After filtration, the first extract is obtained. The filter residue is then mixed with 10,000 ml of ethyl acetate solution and added to the heating furnace. The mixture is heated to boiling for 120 minutes and then filtered again to obtain the second extract. The first and second extracts are then mixed to obtain the β-amyrin extract. In step three above, the β-amyrin extract obtained in step two is placed in a concentration tank, the pressure is adjusted to 97.87 kPa and the temperature is 40℃, and the concentration is carried out under reduced pressure for 120 min. After concentration, an ethyl acetate extract is obtained. In step four above, the ethyl acetate extract obtained in step three is eluted for 5 column volumes with a petroleum ether-ethyl acetate eluent ratio of 50:1 as the first elution to remove impurities. Then, it is eluted again with a petroleum ether-ethyl acetate eluent ratio of 30:1 for 5 column volumes as the second elution. The eluent is collected and placed in a concentration tank. The pressure is adjusted to 96.35 kPa and the temperature is 40°C. The mixture is concentrated under reduced pressure for 120 min to obtain a high-concentration β-amyrin extract. In step five above, the volume ratio of ethyl acetate-ethanol mixed solvent added to the high-concentration β-amyrin extract obtained in step four is 4.5:1. Then the temperature is raised to 58°C, and the mixture is allowed to stand for 150 minutes. After standing, the mixture is filtered to obtain β-amyrin. Example 4:

[0023] A plant rich in β-amyrin, including *Acer sarcodactylis*, wherein the leaves of *Acer sarcodactylis* contain 0.18% to 0.31% β-amyrin.

[0024] A method for extracting β-amyrin includes the following steps: Step 1, drying and sieving; Step 2, extraction and mixing; Step 3, concentrating the extract; Step 4, removing impurities and purifying; Step 5, crystallization. In step one above, 1000g of dried red bean leaves are taken, baked in an oven at 80℃ for 12 minutes, then crushed using a pulverizer, and then sieved through a 20-mesh sieve to obtain red bean leaf powder. In step two above, the leaf powder of red bean is mixed with 10,000 ml of ethyl acetate solution and poured into a heating furnace. The mixture is then heated to boiling for 120 minutes. After filtration, the first extract is obtained. The filter residue is then mixed with 10,000 ml of ethyl acetate solution and added to the heating furnace. The mixture is heated to boiling for 120 minutes and then filtered again to obtain the second extract. The first and second extracts are then mixed to obtain the β-amyrin extract. In step three above, the β-amyrin extract obtained in step two is placed in a concentration tank, the pressure is adjusted to 95.48 kPa and the temperature is 40℃, and the concentration is carried out under reduced pressure for 120 min. After concentration, an ethyl acetate extract is obtained. In step four above, the ethyl acetate extract obtained in step three is eluted for 6 column volumes with a petroleum ether-ethyl acetate eluent ratio of 50:1 as the first elution to remove impurities. Then, it is eluted again with a petroleum ether-ethyl acetate eluent ratio of 30:1 for 6 column volumes as the second elution. The eluent is collected and placed in a concentration tank. The pressure is adjusted to 98.55 kPa and the temperature is 40°C. The mixture is concentrated under reduced pressure for 120 min to obtain a high-concentration β-amyrin extract. In step five above, the volume ratio of ethyl acetate-ethanol mixed solvent added to the high-concentration β-amyrin extract obtained in step four is 4.5:1. Then the temperature is raised to 58°C, and the mixture is allowed to stand for 150 minutes. After standing, the mixture is filtered to obtain β-amyrin.

[0025] The purity of the β-amyrin obtained in the above examples was tested and compared with that of β-amyrin prepared by conventional methods. The results are shown in the table below: Example 1 93.7% Example 2 95.5% Example 3 94.2% Example 4 92.6% Comparative Example 82.3% Based on the above, the advantages of this invention are as follows: It uses leaves of *Adenanthera philoxeroides* as the raw material, which is readily available and reduces raw material costs. The β-amyrin extract is prepared through multiple extractions and mixing, fully extracting β-amyrin from the raw material and reducing its loss during production, thereby increasing the yield. In the purification process, petroleum ether-ethyl acetate with different solvent ratios is used for two elutions, effectively removing impurities from the material and improving the purity of the prepared β-amyrin. Furthermore, it eliminates the need for gel chromatography and liquid chromatography separation, simplifying the preparation method and reducing production difficulty.

[0026] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the invention can be implemented in other specific forms without departing from its spirit or essential characteristics. Therefore, the embodiments should be considered in all respects as exemplary and non-limiting, and the scope of the invention is defined by the appended claims rather than the foregoing description. Thus, all variations falling within the meaning and scope of equivalents of the claims are intended to be included within the present invention. No reference numerals in the claims should be construed as limiting the scope of the claims.

Claims

1. A method for extracting β-amyrin, comprising the following steps: Step 1: Drying and sieving; Step 2: Extraction and mixing; Step 3: Concentrating the extract; Step 4: Purification and removal of impurities; Step 5: Crystallization; Characterized by: In step one above, dried red bean leaves are taken, dried in an oven, then pulverized using a pulverizer, and then sieved through a sieve to obtain red bean leaf powder. In step two above, the leaf powder of red bean is mixed with ethyl acetate solution and poured into a heating furnace. The mixture is then heated to boiling for a period of time. After filtration, the first extract is obtained. The filter residue is then mixed with ethyl acetate solution and added to the heating furnace. The mixture is heated to boiling for a period of time and then filtered again to obtain the second extract. Finally, the first extract and the second extract are mixed to obtain the β-amyrin extract. In step three above, the β-amyrin extract obtained in step two is placed in a concentration tank and concentrated under reduced pressure to obtain an ethyl acetate extract paste. In step four above, the ethyl acetate extract obtained in step three is eluted for the first time with petroleum ether-ethyl acetate to remove impurities, and then eluted a second time with petroleum ether-ethyl acetate. The eluent is collected and concentrated under reduced pressure in a concentration tank to obtain a high-concentration β-amyrin extract. The elution solvent ratio of the petroleum ether-ethyl acetate system in the first elution is 50:1, and 4 to 6 column volumes are eluted. The elution solvent ratio of the petroleum ether-ethyl acetate system in the second elution is 30:1, and 4 to 6 column volumes are eluted. The pressure in the concentration tank is 93.32 to 98.64 kPa, the temperature is 38 to 42°C, and the concentration time under reduced pressure is 110 to 130 min. In step five above, an ethyl acetate-ethanol mixed solvent is added to the high-concentration β-amyrin extract obtained in step four, then the mixture is heated and allowed to stand. After standing, the mixture is filtered to obtain β-amyrin. The volume ratio of ethyl acetate to ethanol in the ethyl acetate-ethanol mixed solvent is 3.5 to 4.8:

1. The mixture is heated to 55 to 60°C and allowed to stand for 120 to 180 minutes.

2. The extraction method for β-amyrin according to claim 1, characterized in that: In step one, the drying temperature is 75-85℃, the drying time is 10-15 minutes, and a 20-mesh sieve is used for screening.

3. The extraction method for β-amyrin according to claim 1, characterized in that: In step two, the mass of the powdered leaves of *Adenanthera philoxeroides* is 950–1050 g, the volume of the ethyl acetate solution mixed with the powdered leaves is 9500–10500 ml, and the boiling time is 110–130 min.

4. The extraction method for β-amyrin according to claim 1, characterized in that: In step two, the volume of the ethyl acetate solution mixed with the filter residue is 9500–10500 ml, and the boiling time is 110–130 min.

5. The extraction method for β-amyrin according to claim 1, characterized in that: In step three, the pressure in the concentration tank is 93.32–98.64 kPa, the temperature is 38–42 °C, and the concentration time under reduced pressure is 110–130 min.

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