Efficient ligustilide extraction method and application thereof

Abstract

The invention discloses an application of ligustilide in preparation of a medicine for improving obesity and complications thereof. The invention further provides an efficient ligustilide extraction method. By taking caenorhabditis elegans as a model organism, it is clear that ligustilide not only can significantly reduce fat accumulation caused by high-fat diet, but also can improve systemic health problems caused by obesity by regulating and controlling various metabolic pathways, such as shortened life, reduced athletic ability, impaired immune function and the like.

Description

Technical Field

[0001] This application relates to the field of pharmaceutical technology, and particularly to a method for highly efficient extraction of ligustilide and its applications. Background Art

[0002] Obesity is a chronic metabolic disease caused by multiple factors, manifested as excessive accumulation and / or abnormal distribution of body fat, resulting in weight gain and potentially leading to a large number of complications, including diabetes, hypertension, coronary heart disease, and impaired immune system function, etc. The continuous increase in the obesity trend has brought a huge burden to human health and the healthcare system, and has become a global public health problem.

[0003] There are many causes of obesity. Among them, excessive intake of high-calorie foods rich in fat and sugar is the main factor leading to obesity. A high-fat diet can change the structure of the intestinal flora and cause excessive accumulation of fat, triggering obesity and related metabolic diseases.

[0004] In recent years, many drugs have been approved for the treatment of obesity. However, the vast majority of weight loss drugs face challenges in terms of efficacy and safety. Especially for systemic problems such as metabolic diseases, decreased immunity, and shortened lifespan caused by obesity, existing drug treatment regimens still lack good effects. Summary of the Invention

[0005] To solve the above technical problems, the first object of the present invention is to provide the application of ligustilide in the preparation of drugs for improving obesity and its complications; the second object of the present invention is to provide a method for highly efficient extraction of ligustilide; the present invention uses Caenorhabditis elegans as a model organism and has clarified that ligustilide can not only significantly reduce fat accumulation caused by a high-fat diet, but also improve systemic health problems caused by obesity, such as shortened lifespan, decreased locomotor ability, and impaired immune function, etc., by regulating multiple metabolic pathways.

[0006] The technical solutions provided by the present invention are as follows: The application of ligustilide in the preparation of drugs for improving obesity and its complications.

[0007] Preferably, improving obesity includes inhibiting fat accumulation.

[0008] Preferably, improving obesity complications includes improving lifespan, improving locomotor ability, and improving intestinal immune deficiency.

[0009] Preferably, the drug includes ligustilide and pharmaceutically acceptable salts, as well as pharmaceutically acceptable excipients.

[0010] Preferably, the dosage form of the drug includes capsules, microcapsules, tablets, granules, dispersible powders, injections, liposomes, oral liquids, intravenous injections, or intramuscular injections; The pharmaceutically acceptable excipients include any one or more of disintegrants, lubricants, emulsifiers, and binders.

[0011] Preferably, the obesity is caused by a high-fat diet.

[0012] A method for highly efficient extraction of ligustilide, comprising the following steps: S1. Grind Chuanxiong, soak and extract it with a mixed solution of petroleum ether and ethanol with a volume ratio of (2 - 8):(8 - 2) at 15 - 25°C for 1 - 4 h, with a material-liquid ratio of 1:(2 - 6) g / ml; then add water for extraction, collect the petroleum ether layer, and concentrate to obtain the volatile oil of Chuanxiong. S2. Use a silica gel column, and elute the volatile oil of Chuanxiong multiple times with a mixed solvent of petroleum ether and ethyl acetate. The volume of the solvent for each elution is equal to the volume of the silica gel column; collect the eluates from the 7th to the 9th time, remove the solvent, and obtain purified ligustilide.

[0013] Preferably, step S1 is specifically: grind Chuanxiong, soak and extract it with a mixed solution of petroleum ether and ethanol with a volume ratio of 6:4 for 3 h, with a material-liquid ratio of 0.25 g / ml; then add water for extraction, collect the petroleum ether layer, and concentrate to obtain the volatile oil of Chuanxiong.

[0014] Ligustilide (CAS NO. 4431-01-0) is a natural terpenoid compound derived from the volatile oil of Chuanxiong, and has an α,β-unsaturated lactone ring in its molecule. In this invention, using Caenorhabditis elegans as a model organism, it is clarified that ligustilide can not only significantly reduce fat accumulation caused by a high-fat diet, but also improve systemic health problems caused by obesity, such as shortened lifespan, decreased locomotor ability, and impaired immune function, etc., by regulating multiple metabolic pathways. This comprehensive effect gives it a unique advantage in the field of anti-obesity.

[0015] Compared with the active substances such as polyphenol extract of Artemisia argyi Levl. et Vant., extract of Notoginseng ginger, and extract of Millettia speciosa Champ. disclosed in the existing literature, the ligustilide provided by this invention has a clear composition and has obvious effects on improving multiple physiological indexes of an obesity model caused by a high-fat diet, especially the intervention and improvement of systemic problems such as metabolic syndrome, decreased immunity, and shortened lifespan caused by obesity.

[0016] This application also provides a method for highly efficient extraction of ligustilide. By response surface analysis (BBD-RSM), the extraction process of ligustilide is optimized, significantly improving the extraction efficiency and purity, and the operation is simple, the cost is low, and it is suitable for industrial production, providing a scientific basis for the large-scale production and industrial application of ligustilide. Description of the Drawings

[0017] To more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the accompanying drawings required for the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only some embodiments recorded in the present application. For those of ordinary skill in the art, without creative efforts, other accompanying drawings can also be obtained based on these drawings.

[0018] Figure 1 It is the experimental result diagram of 13 extraction schemes designed based on BBD-RSM in the embodiments of the present invention; among them, (A) high performance liquid chromatography; (B) regression equation of the peak area concentration curve of LIG standard product.

[0019] Figure 2 It is the experimental result diagram showing that the interaction of extraction parameters is significant in the response surface diagram in the embodiments of the present invention; among them, (A-B) three-dimensional surface diagram (A) and contour diagram (B) of the ratio of petroleum ether to ethanol and extraction time; (C-D) three-dimensional surface diagram (C) and contour diagram (D) of the ratio of petroleum ether to ethanol and liquid-to-material ratio; (E-F) three-dimensional surface diagram (C) and contour diagram (D) of extraction time and the ratio of petroleum ether to ethanol.

[0020] Figure 3 It is the experimental result diagram of the extraction scheme predicted based on BBD-RSM and silica gel column chromatography in the embodiments of the present invention, as well as the thin layer chromatography result diagram; among them, (A) high performance liquid chromatography diagram of the BBD-RSM prediction scheme; (B) thin layer chromatography result diagram; (C) high performance liquid chromatography diagram of purification by silica gel column chromatography method.

[0021] Figure 4 It is the experimental result diagram of ligustilide inhibiting high-fat diet-induced fat accumulation in the embodiments of the present invention; among them, (A) representative pictures of oil red staining (OR) and nile red staining (NR) of nematodes after 7 days of high-fat diet and ligustilide treatment. (B) Quantitative analysis of nile red staining of nematodes. (C) Quantitative analysis of oil red staining of nematodes. (D) Quantitative analysis of the area of nematodes after 7 days of high-fat diet and ligustilide treatment. (E) Quantitative analysis of the body length of nematodes after 7 days of high-fat diet and ligustilide treatment.

[0022] Figure 5 It is the experimental result diagram of ligustilide improving obesity-induced intestinal immune function damage, shortened lifespan and reduced motor ability of nematodes in the embodiments of the present invention; among them, (A) effect of high-fat diet and ligustilide treatment on the lifespan of nematodes. (B) Effect of high-fat diet and ligustilide treatment for 7 days on the movement of nematodes. (C) Effect of high-fat diet and ligustilide treatment for 7 days on the survival time of nematodes under Pseudomonas aeruginosa PA14. (D) Effect of high-fat diet and ligustilide treatment for 7 days on the pharyngeal pumping rate of nematodes. Detailed implementation manners

[0023] To enable those skilled in the art to better understand the technical solutions in this application, the technical solutions in the embodiments of this application will be clearly and completely described below. Obviously, the described embodiments are only part of the embodiments of this application, rather than all of them. Based on the embodiments in this application, all other embodiments obtained by those of ordinary skill in the art without creative efforts belong to the scope protected by this application.

[0024] I. Research on the extraction process of ligustilide 1. Experimental scheme 1.1 Extraction of volatile oil from Ligusticum chuanxiong Ligusticum chuanxiong was dried and pulverized using a high-speed pulverizer, and the powder was obtained by passing through a 40-mesh sieve.

[0025] Among the factors affecting the yield of ligustilide, the volume ratio of petroleum ether to ethanol (2:8, 5:5, 8:2), extraction time (1 - 4 h), and solid-liquid ratio (1:2 - 1:6 g / mL) in the extraction solution were studied and optimized. That is, the above-mentioned extraction solution, extraction time, and solid-liquid ratio were respectively used for soaking extraction at 20 °C. Then, an equal volume of distilled water was added to the extraction solution, and after standing for 30 min, the upper petroleum ether phase was collected, vacuum concentrated at 40 °C, and the volatile oil of Ligusticum chuanxiong was collected.

[0026] 1.2 Silica gel column chromatography The volatile oil of Ligusticum chuanxiong was separated by loading it into a silica gel column. A mixed solvent of petroleum ether and ethyl acetate (volume ratio 99:1) was used as the eluent, and the silica gel column was eluted multiple times. The volume of the solvent for each elution was equal to the volume of the silica gel column. Subsequently, the eluates for each time were collected, and the solvent was removed at 40 °C by rotary evaporation. Finally, high performance liquid chromatography (HPLC) was used to qualitatively and quantitatively analyze each elution fraction to evaluate the main components in the eluate.

[0027] 1.3 Response surface analysis The Box-Behnken method was used to design and optimize the extraction conditions of ligustilide. The Box-Behnken method can achieve similar statistical power with fewer experimental times and show the importance of the interaction between factors. As shown in Table 1, in this procedure, a total of 3 independent factors were established, each factor had three levels, and there were 13 experimental combinations in total. The independent variables of the experiment were the extraction solution ratio (petroleum ether: ethanol), extraction time (min), and solid-liquid ratio (g / mL), and the response (changing with the variable) was the yield of ligustilide. Among them, -1 represented the low level, 0 represented the central level, and 1 represented the high level.

[0028] Table 1 Factors and levels for BBD-RSM process optimization

[0029] 1.4 HPLC Quantitative Analysis High performance liquid chromatography was used to quantitatively analyze Z-ligustilide in the volatile oil. The liquid chromatography analysis instrument was Shimadzu SPD-20A, and the chromatographic column was (Agilent 4.6*250mm Eclipse Plus C18). The ultraviolet detection wavelength was 280 nm. The injection volume was 10 μL, and water and methanol were used as the mobile phase with a flow rate of 1 mL / min. Gradient elution: 0 - 5 minutes, 50% B; 5 - 6 minutes, 50% - 70% B; 6 - 40 minutes, 70% B. For each group of protocols, 5 g of Ligusticum chuanxiong powder was used for extraction, and all volatile oil samples were dissolved in 70 mL of methanol solution and filtered through a 0.22 μm filter membrane before injection. Standard curves were prepared based on standard Z-ligustilide at different concentrations (1, 2, 3, 4, 5, 6 μmol / mL), and the content of Z-ligustilide in the samples was determined according to the peak area.

[0030] 1.5 Thin Layer Chromatography A 10×10 cm silica gel thin layer plate was taken, and a starting line was drawn with a pencil at 1 cm from the bottom, and the sample loading points were marked at equal intervals. A small amount of sample solution was aspirated with a capillary tube and gently dropped at the corresponding sample points. After natural air drying, the silica gel plate was placed in a developing tank. A petroleum ether - ethyl acetate solution with a volume ratio of 99:1 was selected as the developing solution, and the liquid level height was added to 0.5 cm. The developing tank was sealed until the developing solution reached 1 cm from the top of the thin layer plate. The thin layer plate was naturally air dried and developed with iodine vapor.

[0031] 2. Experimental Results 2.1 Establishment of a Process for Highly Efficient Extraction of High-Purity Ligustilide by Response Surface Analysis Based on the BBD-RSM design, 13 groups of protocols (No.1 - No.13) with different factors and 4 groups of repeat experiments (No.14 - No.17) were conducted. High performance liquid chromatography was used to quantitatively analyze the ligustilide extracted from each protocol ( Figure 1 A). The standard curve regression equation between the peak area y (mV) and concentration x (µmol / mL) in the liquid chromatography was determined using the ligustilide standard product ( Figure 1 B). Standard curve equation: Y = 16.739x + 0.0028, R2 = 0.9997. R 2 > 0.99 indicates that the equation has a high degree of fitting to the experimental results and good prediction accuracy. Based on the standard curve regression equation, the extraction yields of ligustilide for each protocol are shown in Table 2.

[0032] Table 2 BBD-RSM Protocols and Extraction Yield Results

[0033] Fitting was performed based on the results of 17 groups of experiments and processed according to the Quadratic model to obtain a ternary quadratic equation with the extraction liquid ratio, extraction time, and liquid-to-solid ratio as independent variables and the extraction rate as the research index:

[0034] Analysis of variance was performed on the ternary quadratic equation. The F value of the quadratic regression model was 43.08 (P < 0.0001), indicating that the regression model was significant (Table 3). By comparing the F values of the extraction liquid ratio, extraction time, and liquid-to-solid ratio factor terms, the significance of these three factors for the research target extraction rate could be directly obtained: extraction time > extraction liquid ratio > liquid-to-solid ratio.

[0035] A three-dimensional surface plot was drawn according to the quadratic regression equation: the interaction surface of the extraction liquid ratio and extraction time ( Figure 2 A - B), the interaction surface of the extraction liquid ratio and liquid-to-solid ratio ( Figure 2 C - D), the interaction surface of the extraction time and extraction liquid ratio ( Figure 2 E - F). From the shape of the surface, a more obvious feature of the surface was that when the proportion of petroleum ether was the lowest (20%) and the extraction time was the shortest (1 h), the extraction rate of ligustilide was the lowest, even less than 1% at this time. And the extraction rate of ligustilide was always not high near this level combination. According to the obtained model and considering the actual operating conditions, the optimal extraction process predicted by BBD - RSM was: the ratio of petroleum ether - ethanol was 6:4, the extraction time was 3 h, and the liquid-to-solid ratio was 250 mg / mL. Under these conditions, the predicted extraction rate of ligustilide was 2.877%.

[0036] Table 3 Analysis of variance of the extraction rate of ligustilide

[0037] To verify the accuracy of the response surface prediction, we collected the volatile oil of Ligusticum chuanxiong Hort. under the optimal extraction process and detected it using liquid chromatography. The results were as Figure 3 shown in A. The liquid chromatography detection results showed that the extraction rate of ligustilide was 2.96%, which was basically consistent with the extraction rate predicted by the response surface method. The volatile oil was separated using silica gel column chromatography, and the eluent was collected and rotary evaporated and concentrated based on the volume of the silica gel column (BV). The eluted components were preliminarily identified using thin layer chromatography. The thin layer chromatography identification results were as Figure 3 shown in B. Ligustilide mainly existed in the eluent of the 7 - 9 BV. The eluted components collected under 7 - 9 BV were mixed and dissolved in methanol to prepare a sample solution of 1 mg / mL, and its purity was further determined by liquid chromatography. The results were as Figure 3As shown in C, the peak area shown by the sample was 86.879, and the purity of ligustilide was calculated to be 98.73% using the standard curve regression equation. The results showed that this extraction process could rapidly obtain high-purity ligustilide.

[0038] II. Experiment on the Activity Effect of Ligustilide 1. Experimental Scheme 1.1 Head Swing Experiment To measure the vitality and health of Caenorhabditis elegans, the head swing of the nematodes was evaluated.

[0039] The nematodes synchronized to the L4 stage were respectively picked into NGM plates with the following four kinds of culture media: (1) NGM plate containing OP50 bacterial solution, (2) NGM plate containing OP50 bacterial solution mixed with ligustilide, (3) NGM plate containing OP50 bacterial solution mixed with egg yolk powder, (4) NGM plate containing OP50 bacterial solution mixed with ligustilide and egg yolk powder. All nematodes were cultured at 20 °C for 7 days. Then, 20 μL of M9 solution was dropped onto the center of the NGM plate, and the recording started 30 seconds after the nematodes were transferred to M9. The number of head swings within 30 s was set as one cycle. This experiment was carried out in a constant temperature environment of 20 °C to ensure that the movement of the nematodes was not affected by temperature.

[0040] 1.2 Lifespan Test The lifespan test of nematodes can effectively reflect the impact on the lifespan of nematodes after drug treatment. The nematodes synchronized to the L4 stage were picked into NGM plates with four different culture media, with 50 nematodes on each plate, and the recording time was day 0. All nematodes were cultured in a constant temperature incubator at 20 °C, and the number of nematodes in each group was recorded every day. Deaths in the first seven days were considered non-natural deaths and would not be recorded. The death criterion was that when the head of the nematode was gently touched and no wriggling or swallowing movement was seen, it was recorded as dead. The daily survival rate = (total number - cumulative number of deaths) / total number of deaths 1.3 Feeding Frequency Experiment In this experiment, the feeding frequency of nematodes was quantified by counting the action frequency of the head pump. The nematodes synchronized to the L4 stage were respectively picked into NGM plates with four different culture media and cultured for 7 days. For the feeding frequency experiment, a multifunctional microscope was used to take videos to record the pharyngeal pump movement frequency of the nematodes within 30 s.

[0041] 1.4 Fat Deposition Experiment The research used Oil Red and Nile Red staining methods to detect fat accumulation in Caenorhabditis elegans. The synchronized L4-stage nematodes were separately picked into NGM plates with the following four types of culture media: (1) NGM plates containing OP50 bacterial solution, (2) NGM plates containing OP50 bacterial solution with mixed ligustilide, (3) NGM plates containing OP50 bacterial solution with mixed egg yolk powder, (4) NGM plates containing OP50 bacterial solution with mixed ligustilide and egg yolk powder. All nematodes were cultured at 20 °C for 7 days. Twenty nematodes from each group were collected into 1.5 mL EP tubes containing M9 and washed three times with M9 solution. After removing the solution, 150 μL of 4% paraformaldehyde was added to each tube and fixed at room temperature for 15 minutes. The fixed nematodes were placed in a -80 °C refrigerator for 2 minutes, and then placed in room temperature water for 1 minute, and this operation was repeated 3 times to break the chitin shell of the nematodes. 1 mL of 60% isopropanol was added to each tube for dehydration for 10 minutes. After removing the solution, 600 μL of Oil Red working solution and Nile Red working solution were added to each tube respectively, and stained for 2 hours in the dark. After staining, it was washed 3 times with M9 buffer containing 0.01% Triton-X100, the nematodes were transferred to an agarose gel pad, and observed and photographed using a fluorescence inverted microscope. Preparation method of Oil Red working solution: 100 mg of Oil Red powder was dissolved in 20 mL of isopropanol solution to prepare an Oil Red stock solution, and then ultrapure water and the Oil Red stock solution were mixed at a ratio of 2:3 (v:v), and shaken on a shaker for 2 hours to prepare the Oil Red working solution. Preparation method of Nile Red working solution: 10 mg of Nile Red powder was added to 20 mL of DMSO and stirred in the dark for 2 hours to obtain a Nile Red stock solution. A 40% isopropanol solution was prepared using isopropanol and ultrapure water, and 6 μL of the Nile Red stock solution was added to each 1 mL of the 40% isopropanol solution to make the Nile Red working solution.

[0042] 2. Experimental Results 2.1 Ligustilide can reduce fat accumulation caused by high-fat diet Caenorhabditis elegans is a classic model organism with a simple structure and has made significant achievements in development, obesity, aging, and metabolic system diseases in the past few decades. Compared with rodent models, Caenorhabditis elegans has many advantages such as a short lifespan, high fecundity, complete cell and developmental lineages, and simple cultivation. The feeding behavior of nematodes is relatively simple and stable, and fat is stored in the form of lipid droplets in the body, and the change process of fat accumulation can be clearly observed. Therefore, based on the fast life cycle of nematodes, easy-to-observe physiological characteristics, and transparent body structure, Caenorhabditis elegans is an ideal research object for constructing an obesity model under high-fat diet.

[0043] Using wild-type Caenorhabditis elegans as a model, we investigated the in vivo effects of ligustilide on the obesity process. The nematodes were cultured in OP50 plates containing different concentrations of ligustilide and egg yolk powder for 7 days, and then the fat accumulation in the nematodes was observed by Oil Red staining and Nile Red staining and quantitatively analyzed.

[0044] The research results are as Figure 4 shown in A - C. Compared with the nematodes in the OP50 group, the fat content of the severely obese group of nematodes treated with a high-fat diet (HFD) increased significantly. Compared with the severely obese group of nematodes, treatment with different concentrations of ligustilide could effectively reduce the fat accumulation in the nematodes, and this effect showed a concentration-dependent enhancement. At the same time, we evaluated the effect of ligustilide on the size of the nematodes. Figure 4 D and Figure 4 E respectively show the area and body length of Caenorhabditis elegans. Compared with the severely obese group of nematodes, treatment with all concentrations of ligustilide could significantly reduce the area and length of the nematodes. Treatment with 50 μM ligustilide could reduce the area and length of the nematodes by about 60% and 30% respectively, and this result was similar to the trend of fat accumulation.

[0045] The results indicate that the natural compound ligustilide derived from Ligusticum chuanxiong can effectively reduce fat accumulation under a high-fat diet and inhibit the occurrence of obesity.

[0046] 2.2 Ligustilide improves the lifespan and immune capacity decline caused by severe obesity We conducted lifespan and intestinal immunity-related experiments on the nematodes treated with ligustilide to further investigate the connection between ligustilide and anti-obesity.

[0047] To evaluate the regulatory ability of ligustilide on the effects of severe obesity, we studied the lifespan index ( Figure 5 A) and locomotion index ( Figure 5 B) of the obese nematodes after treatment with ligustilide. The results showed that compared with the nematodes in the OP50 group, the severely obese nematodes caused by a high-fat diet showed a significant shortening of lifespan and a decline in locomotor ability. However, by feeding the severely obese nematodes with ligustilide, their lifespan and locomotor ability were significantly improved and restored to near the control group level.

[0048] At the same time, nematodes from different groups were simultaneously exposed to the PA14 bacteria environment for culture to further evaluate their intestinal immune capacity ( Figure 5C). The results showed that, compared with the OP50 group, the death rate of nematodes in the obese group increased significantly, suggesting that the intestinal immune function of obese nematodes was severely impaired. After treatment with ligustilide, the death rate of obese nematodes decreased significantly and there was no significant difference from the control group, indicating that ligustilide could effectively improve the intestinal immune deficiency caused by obesity. In addition, we also measured the effect of obesity on feeding ability. The results are as Figure 5 shown in D, obesity affects the feeding ability of nematodes, but ligustilide can improve this situation.

[0049] These results indicate that ligustilide has a significant regulatory effect on various physiological indexes of obese nematodes, including prolonging lifespan, improving locomotor ability, restoring intestinal immune function and enhancing feeding ability. These results provide a scientific basis for the potential application of ligustilide in the field of anti-obesity.

[0050] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to these embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. Application of ligustilide in the preparation of drugs for improving obesity and its complications.

2. The use according to claim 1, characterized in that: Improving obesity includes inhibiting fat accumulation.

3. The use according to claim 1, characterized in that: Improving complications of obesity include improving life expectancy, improving exercise capacity, and improving intestinal immune deficiency.

4. The use according to claim 1, characterized in that: The drug comprises ligustilide and a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

5. The use according to claim 4, characterized in that: The dosage form of the drug includes capsules, microcapsules, tablets, granules, dispersible powders, injections, liposomes, oral solutions, intravenous solutions or intramuscular solutions; The pharmaceutically acceptable excipients include any one or more of disintegrants, lubricants, emulsifiers, and binders.

6. The use according to any one of claims 1 to 5, characterized in that: The obesity is caused by a high-fat diet.

7. A method for efficiently extracting ligustilide, characterized in that: The following steps are involved: S1. Crush the rhizome of Chuanxiong, and use a mixture of petroleum ether and ethanol in a volume ratio of (2-8):(8-2) to soak and extract at 15-25° C. for 1-4 h, with a solid-liquid ratio of 1:(2-6) g / ml; then add water for extraction, collect the petroleum ether layer, and concentrate to obtain volatile oil of the rhizome of Chuanxiong; S2. Using a silica gel column, the volatile oil of Rhizoma Chuanxiong is eluted multiple times with a mixed solvent of petroleum ether and ethyl acetate, and the volume of the solvent for each elution is equal to the volume of the silica gel column; the eluates from the 7th to the 9th time are collected, and the solvent is removed to obtain purified Ligusticum chuanxiong lactone.

8. The method according to claim 7, characterized in that Step S1 is specifically as follows: crushing Chuanxiong, soaking and extracting it in a mixture of petroleum ether and ethanol in a volume ratio of 6:4, the extraction time is 3 hours, and the solid-liquid ratio is 0.25g / ml; then adding water for extraction, collecting the petroleum ether layer, and concentrating it to obtain Chuanxiong volatile oil.

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