阅读说明：本技术 一种微波协同萃取川芎有效物的方法 (Method for microwave synergistic extraction of ligusticum wallichii effective substance ) 是由 陈聪地 杜琳 于海 许青滔 黄荣 刘玉容 张琦 于 2019-12-17 设计创作，主要内容包括：本发明公开了一种微波协同萃取川芎有效物的方法,包括以下步骤：1)将川芎干燥根茎粉碎,过筛,然后加入萃取剂,得到混合液A；2)将混合液A进行梯度式微波萃取,萃取完成获得微波萃取液B；3)将微波萃取液B加入到二氧化碳亚临界萃取装置进行萃取,然后进行解析,得到萃取物；4)纯化步骤：采用色谱硅胶装柱纯化减压,回收溶剂得川芎有效物。本发明采用微波萃取法协同超临界萃取法提取川芎有效物,该方法提取溶剂毒性低、工艺过程简单、产物收率高,成本低,适用于工业化生产。(The invention discloses a method for microwave synergistic extraction of ligusticum wallichii effective substances, which comprises the following steps: 1) pulverizing dried rhizome of rhizoma Ligustici Chuanxiong, sieving, and adding extractant to obtain mixed solution A; 2) carrying out gradient microwave extraction on the mixed solution A, and obtaining microwave extraction liquid B after extraction is finished; 3) adding the microwave extraction liquid B into a carbon dioxide subcritical extraction device for extraction, and then resolving to obtain an extract; 4) a purification step: purifying with chromatographic silica gel column, reducing pressure, and recovering solvent to obtain rhizoma Ligustici Chuanxiong effective component. The invention adopts the microwave extraction method in cooperation with the supercritical extraction method to extract the effective substances of the ligusticum wallichii, and the method has the advantages of low toxicity of the extraction solvent, simple process, high product yield and low cost, and is suitable for industrial production.)

1. A method for microwave synergistic extraction of effective substances of ligusticum wallichii is characterized by comprising the following steps:

1) pulverizing dried rhizome of rhizoma Ligustici Chuanxiong, sieving, and adding 20-25 times of extractant to obtain mixed solution A;

2) carrying out gradient microwave extraction on the mixed solution A, and obtaining microwave extraction liquid B after extraction is finished;

3) supercritical CO₂An extraction step: adding microwave extract B into carbon dioxide subcritical extraction device for extraction, and extracting with CO₂The flow rate is controlled to be 30-35L/h; then, resolving to obtain an extract;

4) a purification step: loading chromatographic silica gel into a column with a diameter-height ratio of 1:0.5-1:3, dissolving the extract with methanol, adding 1-2 times of solvent A for elution to obtain eluent I, discarding the eluent, eluting with 4-7 times of solvent B to obtain eluent II, and recovering solvent from the eluent II at 35-40 deg.C under reduced pressure to obtain rhizoma Ligustici Chuanxiong effective component.

2. The method as claimed in claim 1, wherein the extractant in step 1) comprises chlorinated gallbladder and ethanol at a molar ratio of 1:2-1: 4.

3. The method for microwave synergistic extraction of ligusticum wallichii effective substance according to claim 1 or 2, wherein the microwave extraction in the step 2) is divided into two stages: the first stage, the mixed solution A is subjected to microwave extraction for 8-10min at the temperature of 40-50 ℃, the microwave frequency of 2000-2200 MHz and the power of 150-180W; in the second stage, the temperature, time and power of the extraction are the same as those in the first stage, and the extraction frequency is 2800-3000 MHz.

4. The method as claimed in claim 3, wherein the extraction pressure in step 3) is 25-30MPa and the extraction temperature is 40-55 ℃.

5. The method as claimed in claim 3, wherein the desorption pressure in step 3) is 6-9MPa and the desorption temperature is 25-38 ℃.

6. The method as claimed in claim 1, wherein the step 3) of supercritical CO extraction₂Continuously pumping entrainer in the extraction process, wherein the addition amount of the entrainer is 6-10% of the mass of the extraction liquid B; the entrainer comprises acetone, ethanol and butyl acrylate, and the adding proportion is 1:3-4:1 by weight.

7. The method as claimed in claim 6, wherein the step 3) of supercritical CO extraction₂The polyglycerol fatty acid ester is used as additive in the extraction process, and the dosage of the additive is 0.2-0.3% of the volume of the extraction device.

8. The method as claimed in claim 7, wherein the step 3) of supercritical CO extraction₂Vitamin C which accounts for 0.1-1% of the volume of the extraction device is also added as a stabilizer in the extraction process.

9. The method of claim 1, wherein the linear velocity of the eluting solvent in step 4) is 0.2-8 cm-min^-1。

10. The method for microwave synergistic extraction of rhizoma Chuanxiong active ingredients as claimed in claim 1, wherein the solvent A in step 4) is one or more of petroleum ether, n-hexane, n-heptane, and ethyl acetate; the solvent B is one or a mixture of ethyl acetate, dichloromethane and chloroform.

Technical Field

The invention relates to the technical field of extraction of effective components of traditional Chinese medicinal materials, in particular to a method for microwave synergistic extraction of ligusticum wallichii effective substances.

Background

The rhizoma Ligustici Chuanxiong is dried rhizome of Ligusticum chuanxiong Hort of Umbelliferae, and has effects of promoting blood circulation, activating qi-flowing, dispelling pathogenic wind and relieving pain. The rhizoma ligustici wallichii is a traditional Chinese medicine in China and is also a famous medicinal material in Chuan producing area, and according to statistics, the rhizoma ligustici wallichii is produced in domestic years by 5 multiplied by 10⁶～6×10⁶In 564 finished prescription preparations and single preparation collected in the section I of Chinese pharmacopoeia (2005 edition), 85 kinds of ligusticum wallichii are used, which accounts for about 15% of Chinese patent medicines collected in the pharmacopoeia. Ligusticum wallichii contains various lactones, alkaloids, phenols, volatile oils and other compounds, and in recent years, the research shows that the polysaccharide of Ligusticum wallichii has the function of inhibiting the growth of cancer cells. Therefore, the method has wide prospect for researching the functions and the application of the ligusticum wallichii polysaccharide. However, the active components in the ligusticum wallichii are complex, have low content and are difficult to enrich.

The traditional extraction method mainly comprises a water vapor distillation method, a percolation method, a solvent extraction method and the like. The method introduced in journal "journal of pharmacy" at page 482 of phase 6 in 1988 and "journal of Chinese materia medica" at page 147 of phase 3 in 1996 by steam distillation method is as follows: firstly, the raw materials are crushed, then the crushed raw materials are placed in a steam extractor for distillation and extraction for a period of time, finally, the extracted oil is dried and dehydrated by anhydrous sodium sulfate, and the yield of the completely extracted oil is between 0.05 and 1.20 percent. The method has the advantages of high extraction temperature, easy damage to some effective components, extremely low extraction rate and high production cost; meanwhile, the steam distillation process can cause the hydrolytic deterioration, rearrangement, oxidation, polymerization, resinification and the like of volatile components, so that the volatile components are not completely the same as the original components of the ligusticum wallichii, and the actually measured components and content of the sample are not in accordance with the actual results. The percolation method has low efficiency and is difficult to be produced in modern times. The solvent extraction method mainly focuses on solvent extraction and extraction processes, and further purification processes are reported less.

In order to solve the defects in the prior art, the invention provides a method for extracting ligusticum wallichii effective substances by microwave synergy.

Disclosure of Invention

In view of the above, the invention provides a method for extracting ligusticum wallichii effective substances by microwave synergistic extraction, and the method adopts the microwave extraction method and the supercritical extraction method to extract the ligusticum wallichii effective substances, has the advantages of low toxicity of extraction solvent, simple process, high product yield and low cost, and is suitable for industrial production.

In order to achieve the purpose, the invention adopts the following technical scheme:

a method for microwave synergistic extraction of rhizoma Ligustici Chuanxiong effective components comprises the following steps:

1) pulverizing dried rhizome of rhizoma Ligustici Chuanxiong, sieving, and adding 20-25 times of extractant to obtain mixed solution A;

2) carrying out gradient microwave extraction on the mixed solution A, and obtaining microwave extraction liquid B after extraction is finished;

3) supercritical CO₂An extraction step: adding microwave extract B into carbon dioxide subcritical extraction device for extraction, and extracting with CO₂The flow rate is controlled to be 30-35L/h; then, resolving to obtain an extract;

4) a purification step: loading chromatographic silica gel into a column with a diameter-height ratio of 1:0.5-1:3, dissolving the extract with methanol, adding a dissolving solution with an amount of 2-4 times of the column volume, adding a solvent A with an amount of 1-2 times of the column volume for elution to obtain an eluent I, discarding the eluent I, eluting with a solvent B with an amount of 4-7 times of the column volume to obtain an eluent II, and recovering the solvent from the eluent II under reduced pressure at 35-40 ℃ to obtain the ligusticum wallichii effective substance.

Preferably, the extractant in the step 1) consists of chlorinated gallbladder and ethanol according to a molar ratio of 1:2-1: 4.

Preferably, the microwave extraction in step 2) is divided into two stages: the first stage, the mixed solution A is subjected to microwave extraction for 8-10min at the temperature of 40-50 ℃, the microwave frequency of 2000-2200 MHz and the power of 150-180W; in the second stage, the temperature, time and power of the extraction are the same as those in the first stage, and the extraction frequency is 2800-3000 MHz.

Preferably, the extraction pressure in the step 3) is 25-30MPa, and the extraction temperature is 40-55 ℃.

Preferably, the desorption pressure in the step 3) is 6-9MPa, and the desorption temperature is 25-38 ℃.

Preferably, the step 3) of supercritical CO₂The extraction process is continuously pumped with the carrierThe addition amount of the entrainer is 6-10% of the mass of the extract B; the entrainer comprises acetone, ethanol and butyl acrylate, and the adding proportion is 1:3-4:1 by weight.

Preferably, the step 3) of supercritical CO₂The polyglycerol fatty acid ester is used as additive in the extraction process, and the dosage of the additive is 0.2-0.3% of the volume of the extraction device.

Preferably, the step 3) of supercritical CO₂Vitamin C which accounts for 0.1-1% of the volume of the extraction device is also added as a stabilizer in the extraction process.

Preferably, the linear velocity of the elution solvent in the step 4) is 0.2-8 cm-min^-1。

Preferably, the solvent A in the step 4) is one or more of petroleum ether, n-hexane, n-heptane and ethyl acetate; the solvent B is one or a mixture of ethyl acetate, dichloromethane and chloroform.

Preferably, the crushed materials in the step 1) are sieved by a 50-100 mesh sieve.

According to the technical scheme, compared with the prior art, the invention has the following beneficial effects:

1. the invention adopts reasonable extractant to pretreat the ligusticum wallichii, thereby laying a good foundation for efficiently extracting effective substances in the ligusticum wallichii in subsequent steps;

2. although the supercritical extraction method and the microwave method are the most commonly used extraction methods at present, the methods have advantages and disadvantages, and the extraction rate of the invention is improved by 50-60% compared with the single extraction method by adopting the microwave extraction method and the supercritical extraction method to extract the effective substances of the ligusticum wallichii.

3. The invention adopts a gradient extraction method in the microwave extraction process, adds entrainer, additive and stabilizer which are not disclosed in the prior art in the subcritical extraction process, determines reasonable proportion through a large number of experiments, shortens the extraction time of the whole process, improves the yield and reduces the cost.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.