阅读说明：本技术 一种高提取率的黄芪甲苷提取方法 (Method for extracting astragaloside with high extraction rate ) 是由 岳露凯 张元坤 于 2021-07-13 设计创作，主要内容包括：发明公开了一种高提取率的黄芪甲苷提取方法,包括如下步骤：将冷冻后的黄芪超微粉与碱性甲醇水溶液混匀,加压浸透,然后加水,减压提取得到提取液,纯化得到黄芪甲苷。本发明黄芪甲苷的提取率高,黄芪甲苷的纯度高。(The invention discloses a method for extracting astragaloside with high extraction rate, which comprises the following steps: mixing the frozen radix astragali superfine powder with alkaline methanol aqueous solution, pressurizing and soaking, adding water, extracting under reduced pressure to obtain extractive solution, and purifying to obtain astragaloside IV. The method has high extraction rate of the astragaloside and high purity of the astragaloside.)

1. A method for extracting astragaloside IV with high extraction rate is characterized by comprising the following steps: mixing the frozen radix astragali superfine powder with alkaline methanol aqueous solution, pressurizing and soaking, adding water, extracting under reduced pressure to obtain extractive solution, and purifying to obtain astragaloside IV.

2. The method for extracting astragaloside IV with high extraction rate as claimed in claim 1, wherein the particle size of the radix astragali ultra-fine powder is 300-400 mesh; preferably, the freezing temperature is less than or equal to-20 ℃, and the freezing time is 8-12 h.

3. The method for extracting astragaloside IV with high extraction rate according to claim 1 or 2, wherein the temperature of the alkaline methanol aqueous solution is 40-50 ℃; preferably, the pH of the aqueous alkaline methanol solution is 10-11; preferably, the volume fraction of methanol in the basic aqueous methanol solution is 75-80%.

4. The method for extracting astragaloside IV with high extraction rate according to any one of claims 1-3, wherein the weight volume (g/ml) ratio of the astragalus ultra-micro powder to the alkaline methanol aqueous solution is 1: 3-5.

5. The method for extracting astragaloside IV with high extraction rate according to any one of claims 1-4, wherein the pressure for pressure impregnation is 11-13 MPa; preferably, the time for pressurizing and soaking is 60-90 min; preferably, the temperature for pressure impregnation is 40-50 ℃.

6. The method for extracting astragaloside IV with high extraction rate according to any one of claims 1-5, wherein the temperature of water is 70-80 deg.C; preferably, the weight ratio of the astragalus membranaceus superfine powder to water is 1: 10-12.

7. The method for extracting astragaloside IV with high extraction rate according to any one of claims 1-6, wherein the pressure for the extraction under reduced pressure is 0.03-0.04 MPa; preferably, the extraction time under reduced pressure is 20-25 min; preferably, the extraction is performed 1-3 times under reduced pressure.

8. The method for extracting astragaloside IV with high extraction rate according to any one of claims 1-7, characterized in that the purification method comprises the following specific steps: eluting the extractive solution with HPD300 resin column to obtain eluate, concentrating the eluate to obtain astragaloside IV crude product, and recrystallizing to obtain astragaloside IV.

9. The method for extracting astragaloside IV with high extraction rate as claimed in claim 8, wherein the column elution is carried out with 68-72% by volume of ethanol aqueous solution, and the amount of ethanol aqueous solution is 4-5 times the column volume.

10. The method for extracting astragaloside IV with high extraction rate as claimed in claim 8, wherein the recrystallization solution is methanol.

Technical Field

The invention relates to the technical field of traditional Chinese medicine extraction, in particular to a method for extracting astragaloside IV with high extraction rate.

Background

Astragalus belongs to the genus Astragalus of the family Leguminosae, which is classified into Astragalus mongholicus and Astragalus membranaceus, and is mainly distributed in Heilongjiang, Jilin, inner Mongolia and other provinces, and is called the top grade of medicine. Huangmao is slightly warm, sweet in nature, spleen meridian entered lung, beneficial to induce diuresis to expel toxin, tonify qi to consolidate superficies, expel pus, heal wound and promote granulation.

Radix astragali contains Astragalus polysaccharides, saponins (astragalosides I-VIII), flavonoids (kaempferol, rhamnitrin, perkin, flavonoid glycoside, etc.), amino acids (aspartic acid, canavanine, alanine, proline, r-aminobutyric acid, arginine, etc.), and other trace elements (iron, manganese, zinc, etc.). The astragalus polysaccharides and saponins are the main active substances in astragalus, and the astragalus saponins (I-VIII) are the main saponins in astragalus. The content of astragaloside in radix astragali is an important index for quality control of medicinal materials. Astragaloside IV is beneficial to the positive inotropic action of the heart and is one of the important effective components for reducing blood pressure.

The astragaloside IV is astragaloside IV in the triterpene saponin of radix astragali, the content of astragaloside IV in radix astragali is very low, generally 0.04-0.14%, the ingredients in radix astragali are more, and the separation and extraction are very difficult. At present, the extraction method of astragaloside IV generally adopts a water extraction method to obtain an astragalus extract, and then adopts methods such as a n-butanol extraction method, a macroporous adsorption resin method and the like to purify and separate saponin monomers, but the method has the advantages of long period, complex operation, higher production cost and lower yield.

Disclosure of Invention

Based on the technical problems in the background art, the invention provides the method for extracting the astragaloside with high extraction rate.

The invention provides a method for extracting astragaloside with high extraction rate, which comprises the following steps: mixing the frozen radix astragali superfine powder with alkaline methanol aqueous solution, pressurizing and soaking, adding water, extracting under reduced pressure to obtain extractive solution, and purifying to obtain astragaloside IV.

Preferably, the grain diameter of the astragalus root superfine powder is 300-400 meshes.

Preferably, the freezing temperature is less than or equal to-20 ℃, and the freezing time is 8-12 h.

Preferably, the temperature of the basic aqueous methanol solution is 40-50 ℃.

Preferably, the pH of the aqueous alkaline methanol solution is 10-11.

The pH can be adjusted with sodium hydroxide or the like.

Preferably, the volume fraction of methanol in the basic aqueous methanol solution is 75-80%.

Preferably, the weight-volume (g/ml) ratio of the astragalus membranaceus superfine powder to the alkaline methanol aqueous solution is 1: 3-5.

Preferably, the pressure for pressure impregnation is 11-13 MPa.

Preferably, the time for pressure soaking is 60-90 min.

Preferably, the temperature for pressure impregnation is 40-50 ℃.

Preferably, the temperature of the water is 70-80 ℃.

Preferably, the weight ratio of the astragalus membranaceus superfine powder to water is 1: 10-12.

Preferably, the pressure of the reduced pressure extraction is 0.03-0.04 MPa.

Preferably, the extraction time under reduced pressure is 20-25 min.

Preferably, the extraction is performed 1-3 times under reduced pressure.

Preferably, the specific steps of the purification method comprise: eluting the extractive solution with HPD300 resin column to obtain eluate, concentrating the eluate to obtain astragaloside IV crude product, and recrystallizing to obtain astragaloside IV.

Preferably, the column elution is carried out with 68-72% by volume of an aqueous ethanol solution, which is used in an amount of 4-5 column volumes.

Preferably, the recrystallization solution is methanol.

Has the advantages that:

the astragalus mongholicus superfine powder is frozen at a low temperature and then is uniformly mixed with an alkaline methanol aqueous solution at a temperature of 40-50 ℃, fibers, cell walls and the like in the astragalus mongholicus are subjected to extremely-temperature-difference rapid heat treatment and can be broken and crushed, then the cell walls are further crushed by pressurizing and soaking, the alkaline methanol aqueous solution is promoted to enter the inside of cells, so that the extraction of effective components is promoted, in addition, the soaking in an alkaline environment can promote the astragalosides I-II, and the deacetylation is converted into astragaloside, so that the content of the astragaloside is improved; then adding water with a certain temperature, immediately reducing the pressure to boil the solution, thereby extracting and obtaining astragaloside IV; by the previous shock heating and pressurizing soaking treatment, the astragaloside IV can be more easily and completely extracted from broken cells, so that the extraction rate is improved; then eluting by HPD300 resin column, recrystallizing to obtain high purity astragaloside IV; the alkaline methanol aqueous solution and water used for extraction are less in dosage, so that the dosage of the solvent during column elution can be greatly reduced, the purification time is shortened, the dosage of the solvent is reduced, and the cost is saved.

Detailed Description

The technical solution of the present invention will be described in detail below with reference to specific examples.

The astragalus powder of the following examples 1 to 3 and comparative examples 1 to 2 is a product of the same lot purchased from the same manufacturer, and has a particle size of 60 mesh.

Example 1

A method for extracting astragaloside IV with high extraction rate comprises the following steps:

weighing radix astragali powder, micronizing to obtain radix astragali superfine powder with particle size of 320 meshes, weighing 5g radix astragali superfine powder, freezing at-20 deg.C for 10h, taking out, immediately mixing with 20ml methanol aqueous solution with pH of 10.5, temperature of 45 deg.C and volume fraction of 77%, transferring to a reaction kettle, adjusting pressure to 12Mpa, soaking at 45 deg.C for 80min, adding 75 deg.C water, immediately reducing pressure to 0.03MPa, extracting for 2 times (water dosage is 55ml each time, and extraction time under reduced pressure is 23min each time), and mixing extractive solutions;

eluting with HPD300 resin column (using 70 vol% ethanol water solution for column elution, the amount of ethanol water solution is 5 times of column volume) to obtain eluate, concentrating the eluate under reduced pressure to obtain astragaloside IV crude product, and recrystallizing with methanol to obtain astragaloside IV.

Example 2

A method for extracting astragaloside IV with high extraction rate comprises the following steps:

weighing radix astragali powder, carrying out superfine grinding to obtain radix astragali superfine powder with the particle size of 300 meshes, weighing 5g of radix astragali superfine powder, freezing at-20 ℃ for 12h, taking out, immediately mixing with 25ml of 75% methanol aqueous solution with the pH of 10, the temperature of 50 ℃ and the volume fraction of 75%, transferring to a reaction kettle, regulating the pressure to 11Mpa, soaking at 50 ℃ for 60min, then adding 50ml of water with the temperature of 80 ℃, immediately reducing the pressure to 0.04MPa, extracting for 1 time, wherein the time of each reduced pressure extraction is 25min, and combining the extracting solutions;

eluting with HPD300 resin column (using 68% ethanol water solution for column elution, the amount of ethanol water solution is 5 times of column volume) to obtain eluate, concentrating the eluate under reduced pressure to obtain astragaloside IV crude product, and recrystallizing with methanol to obtain astragaloside IV.

Example 3

A method for extracting astragaloside IV with high extraction rate comprises the following steps:

weighing radix astragali powder, performing superfine grinding to obtain radix astragali superfine powder with the particle size of 400 meshes, weighing 5g of radix astragali superfine powder, freezing at-20 ℃ for 8h, taking out, immediately mixing with 15ml of methanol aqueous solution with the pH value of 11, the temperature of 40 ℃ and the volume fraction of 80%, transferring to a reaction kettle, regulating the pressure to 13Mpa, soaking at 40 ℃ for 90min, then adding water with the temperature of 70 ℃, immediately reducing the pressure to 0.03MPa, extracting for 3 times (the water consumption is 60ml each time, and the time for extracting under reduced pressure is 20min each time), and combining the extracting solutions;

eluting with HPD300 resin column (with 72% ethanol water solution at 4 times of column volume) to obtain eluate, concentrating the eluate under reduced pressure to obtain astragaloside IV crude product, and recrystallizing with methanol to obtain astragaloside IV.

Comparative example 1 (Water extraction method)

An extraction method of astragaloside IV comprises the following steps:

weighing 5g radix astragali powder with particle size of 60 mesh, decocting with water for 2 times (55 ml for each time, 1.5 hr for each time), and filtering to obtain extractive solution;

eluting with HPD300 resin column (using 70 vol% ethanol water solution for column elution, the amount of ethanol water solution is 5 times of column volume) to obtain eluate, concentrating the eluate under reduced pressure to obtain astragaloside IV crude product, and recrystallizing with methanol to obtain astragaloside IV.

Comparative example 2 (internal extraction under reduced pressure)

An extraction method of astragaloside IV comprises the following steps:

weighing radix astragali powder, micronizing to obtain radix astragali superfine powder with particle size of 320 meshes, weighing 5g radix astragali superfine powder, mixing with 77% methanol water solution 20ml, soaking for 80min, transferring to a reaction kettle, adding 75 deg.C water, immediately reducing pressure to 0.03MPa, extracting for 2 times (each time water amount is 55ml, each time reduced pressure extraction time is 23min), and mixing extractive solutions;

weighing 5g radix astragali powder with particle size of 60 mesh, decocting with water for 2 times (55 ml for each time, 1.5 hr for each time), and filtering to obtain extractive solution;

eluting with HPD300 resin column (using 70 vol% ethanol water solution for column elution, the amount of ethanol water solution is 5 times of column volume) to obtain eluate, concentrating the eluate under reduced pressure to obtain astragaloside IV crude product, and recrystallizing with methanol to obtain astragaloside IV.

Experiment of

Detecting the content of astragaloside IV in the extractive solution according to reference external standard method, detecting the purity of finally obtained astragaloside IV, and calculating the extraction rate of astragaloside IV in the extractive solution and the yield of finally obtained astragaloside IV product.

The detection method comprises the following steps:

high performance liquid chromatography (Agilent1200, usa), evaporative light scattering detector ELSD (2000ES, alttech, usa), rotary evaporator (RE-52AA, shanghai);

a chromatographic column: agilent XDB-C18(150 mm. times.4.6 mm, 5 μm), mobile phase: acetonitrile-water 32:68(v/v), flow rate 1.0ml/min, column temperature 30 ℃, sample size 5 μ L, ELSD parameters: the drift tube temperature was 105 ℃ and the N2 flow rate was 2.9 mL/min.

The results are shown in Table 1.

TABLE 1 test results

As can be seen from the above table, the method has high extraction rate of the astragaloside and the prepared astragaloside has high purity.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.