AU2020102524A4

AU2020102524A4 - Production method of decursin

Info

Publication number: AU2020102524A4
Application number: AU2020102524A
Authority: AU
Inventors: Xiangqian Liu
Original assignee: Changsha Broad Ocean Bio Science And Technique Co Ltd
Current assignee: Changsha Broad-Ocean Bio-Science And Technique Co Ltd
Priority date: 2020-09-30
Filing date: 2020-09-30
Publication date: 2020-11-19
Anticipated expiration: 2028-09-30

Abstract

The invention relates to a production method of decursin, in particular to a technical method for producing decursin from Angelica decursiva by ultrasonic extraction. The method takes Angelica decursiva as a raw material and includes the steps of raw material crushing, extraction, refining and the like, and is characterized in that the extraction step includes the step of carrying out ultrasonic extraction under the temperature of 40-70 °C and the frequency of 40-60 KHz by using a polar organic solvent as a solvent. The production method of the decursin is not only low in temperature and short in time, but also obtains unexpected high extraction efficiency. Meanwhile, in the technical process of the invention, the temperature is lower, and effective components are not damaged easily, so that post-treatment processes are facilitated and product is high in purity.

Description

Production method of decursin

TECHNICAL FIELD

[01] The invention relates to a production method of decursin, in particular to a technical method for producing decursin from Angelica decursiva by ultrasonic extraction. The method belongs to the field of chemistry of Chinese materia medica.

BACKGROUND

[02] Angelica decursiva is the dried root of umbelliferae Angelica decursiva (Miq.) Franch and a perennial ratoon herb plant, which has the effects of dispelling wind and clearing, reducing qi and eliminating phlegm and the like, is mainly produced in Hunan, Jiangxi, Anhui or the like, is widely distributed in China, and is also a common traditional Chinese medicine in clinic application. It has effects in dispelling wind, clearing away heat, lowering qi, and eliminating phlegm; and can be used for treating symptom such as cough caused by wind-heat and excessive phlegm and used for eliminating dampness and clearing away heat. Modem research shows that the plant contains volatile oil and chemical components such as coumarin and the like, for example, the plant mainly includes nodakenin, nodakenetin, decursin, cynanolide and the like. The molecular formula of the decursin is C19H2005, the melting point is 110 112 DEG c, and [a]d1 5: + 172 (c=7,CHCL3). The research results show that the decursin has obvious effects on treating leukemia, diabetes, hypertension and reducing nephrotoxin, and can inhibit helicobacter pylori which are closely related to diseases such as gastric ulcer, duodenal ulcer, gastritis, gastric cancer and the like.

0

Decursin

[03] Extraction methods of decursin disclosed in the prior art are generally carried out by using diethyl ether as a solvent. These methods include a thermal extraction method such as reflux extraction or Soxhlet extraction and the like, and a cold leaching method. Diethyl ether has a low boiling point and is easy to volatilize and explode, so that the method is not easy for industrial large-scale production. Decursin is unstable and is easy to be damaged when being heated and extracted at a high temperature, so that the yield and the product content are reduced. The cold leaching extraction method is characterized by low extraction efficiency, long process time and incapability of being suitable for industrial production. Therefore, it is very meaningful to provide a safe and efficient method for producing decursin.

SUMMARY

[04] In order to overcome the defects in the prior art, the invention provides a production technology of decursin, which is safe, more effective and suitable for industrial production. The technical scheme of the invention is as follows:

[05] The invention relates to a production method of decursin, which takes Angelica decursiva as a raw material and includes the steps of raw material crushing, extraction, refining and the like. The method is characterized in that the extraction step includes the step of carrying out ultrasonic extraction under the temperature of 40-70 °C and the frequency of 40-60 KHZ by taking a polar organic solvent as a solvent.

[06] The Angelica decursiva used as a raw material can be fed by using roots, stems, leaves, fruits and the like of the Angelica decursiva, wherein the roots of Angelica decursiva are preferably used for feeding. Before the raw material is fed, certain pretreatment is needed. In general, the raw materials are crushed into proper powder to for feeding and extraction. According to the invention, the Angelica decursiva raw material is preferably crushed into powder of 20-60 meshes for feeding.

[07] As the extraction solvent in the method of the present invention, a polar organic solvent is selected as the extraction solvent, such as methanol, ethanol or acetone, or a mixed solvent composed of the polar organic solvent can be selected as the extraction solvent, or an aqueous mixed solvent containing a small amount of water is selected as the extraction solvent, and the water content thereof should be less than %. As a preferred embodiment of the present invention, 90%-95% ethanol is selected as the solvent for ultrasonic extraction.

[08] According to the production method disclosed by the invention, the refining step includes the following steps of: extracting an extract obtained by ultrasonic extraction with chloroform or a solvent which has a polarity similar to that of the chloroform; and drying the obtained extract at low temperature and then carrying out column chromatography separation.

[09] Among them, the "solvent which has a polarity similar to that of chloroform" may be a single solvent which a polarity similar to that of chloroform, such as an organic solvent such as dichloromethane, or a mixed solvent which has a polarity similar to that of chloroform, such as a mixed solvent of petroleum ether or n-hexane and a solvent such as ethyl acetate or acetone in an appropriate ratio. Suitable solvents may be selected as needed for those skilled in the art.

[010] In the production method described above, it is preferable to use a low temperature drying method for drying the extract. For example, a low-temperature drying method such as a reduced-pressure drying method, a spray drying method, and a microwave drying method can be used. As a preferred embodiment of the present invention, a microwave drying method is preferably employed.

[011] Specifically, in the production method, conventional traditional Chinese medicine separation approaches such as column chromatography, solvent extraction and the like can be selected for refining or purifying the extract. According to the present invention, column chromatography is preferably used for separation. There is no particular requirement for stationary and mobile phases. Those skilled in the art can select suitable separation materials for separation as needed. As a specific implementation of the present invention, silica gel column chromatography is preferred, and a mixed solvent of n-hexane and ethyl acetate is used as the eluent for gradient elution.

[012] As a preferred embodiment of the present invention, the above production method includes crushing Angelica decursiva into 20-60 meshes of powder, carrying out ultrasonic extraction under the temperature of 40-70 °C and the frequency of 40 KHz for 30 min with 90-95% ethanol, carrying out extraction for twice, mixing extractive solutions, carrying out filtration, recovering ethanol from the filtrate, carrying out dilution with water, carrying out extraction with chloroform for 3-5 times, mixing polar layers, recovering chloroform, carrying out microwave drying at a low temperature, carrying out chromatography on a silica gel column, carrying out gradient elution with a mixed solvent of n-hexane and ethyl acetate, and collecting decursin.

[013] The production method disclosed by the invention further includes, if necessary, the step of further purifying the obtained decursin. For example, the decursin prepared by the method is subjected to a column chromatography or recrystallization step, so that the purity of the product is improved to the required sample purity. Specifically, it is obvious that conventional purification steps are carried out according to the material properties of the decursin and the means of separating and refining the Chinese herbal medicines. Preferably, the appropriate manners of column chromatography and recrystallization are used for carrying out conventional steps of purification and completing the refining of a crude product, which belongs to the general technology in the field.

[014] In order to illustrate the effect of the invention, the extraction efficiency and the product purity of the decursin are also measured. A high performance liquid chromatography (HPLC) is used for detecting the decursin. The detection conditions of the HPLC are as follows:

[015] Preparing a mobile phase:

[016] (1) After being diluted by 10 times by thephosphoric acid-sodium dihydrogen phosphate buffer solvent with the pH value of 5, the mixture is mixed with acetonitrile according to the volume ratio of 1: 1;

[017] (2) Dodecyl sulfate is added to the mixture at a ratio of the mixture to the dodecyl sulfate of 1,000 ml (1) to 2.88 g.

[018] Detection conditions:

[019] The column temperature is 35 °C and the flow rate is 1.2 ml/min. The detection wavelength is 280 nm, wherein the ODS column is subject to 250x4.6 mm 5

pm.

[020] According to the invention, as found by chance, under selection of the ultrasonic extraction method and under a certain extraction condition, the process for producing the decursin from the Angelica decursiva raw material is not only low in temperature and short in time, but also obtains unexpected high extraction efficiency. Meanwhile, in the technical process of the invention, the temperature is lower, and the effective components are not damaged easily, so that a post-treatment process is facilitated, and the purity of the product is high.

DESCRIPTION OF THE INVENTION

[021] The following examples are provided merely to illustrate or explain the present invention and should not be understand to form limits or limit the scope of protection of the appended claims of the invention.

[022] EMBODIMENT 1

[023] Dried Angelica decursiva roots of 1KG are taken, crushed to 60 meshes, and then subjected to ultrasonic extraction for 30 minutes under the temperature of 50 °C and the frequency of 60 KHZ, wherein the solvent is 95% ethanol, extraction is carried out for twice, extracting solutions are mixed, filtration is carried out, the ethanol is recovered, 200ML of water is added for dilution, extraction is carried out for four times with 1000ML of chloroform, the chloroform is recovered, and microwave low temperature drying is carried out till dry paste of 30g is obtained.

[024] The dry past is mixed uniformly with 60 g of silica gel and then loaded on a silica column (the amount of the silica gel is 1000 g), gradient elution is carried out by using n-hexane and ethyl acetate in volume ratios of 9:1, 5:1, 3:1, 2:1, and 1:1, the part (3:1) of the decursin is collected, a solvent is recovered, then silica gel column chromatography is carried out, elution is carried out by using n-hexane and ethyl acetate with ratios of 3:1 and 2:1, the part (2: 1) is collected, the solvent is recovered, about 6 g of the decursin is obtained, and the content is 98.6% as detected by an HPLC method.

[025] EMBODIMENT 2 (Contrast Experiment)

[026] Dried Angelica decursiva root medical materials of a proper amount are taken and crushed into powder of 40 meshes.

[027] Extracting a): taking a proper amount of the powder, precisely weighing the powder, ultrasonically extracting the powder for 30 minutes at 55 °C and the frequency of 55 KHZ, wherein the solvent is 95% ethanol, extracting the powder twice, mixing the extracting solutions, filtering the extracting solutions, dissolving the extracting solutions by using methanol, fixing the volume to 100ml, passing through a 0.45pm filter membrane to serve as a sample solution, and measuring the sample solution according to the high performance liquid chromatography method;

[028] Extracting b): taking a proper amount of the powder, precisely weighing, extracting twice with 95% ethanol for 2 hours each time, mixing the extractive solutions, filtering, dissolving with methanol, fixing the volume to 100ml, passing through a 0.45pm filter membrane to serve as a sample solution, and measuring the solution according to the high performance liquid chromatography method.

[029] Results: the content of the decursin in the medicinal materials measured by the extraction method a (the method of the invention) is 0.679%, the content of the prednisone in the medicinal materials measured by the extraction method b (the comparison method) is only 0.0786%, which is almost one tenth of that of the method of the invention; and moreover, the extraction time is long, and the extraction efficiency difference is remarkable.

[030] Although the invention has been described with reference to specific examples, it will be appreciated by those skilled in the art that the invention may be embodied in many other forms, in keeping with the broad principles and the spirit of the invention described herein.

[031] The present invention and the described embodiments specifically include the best method known to the applicant of performing the invention. The present invention and the described preferred embodiments specifically include at least one feature that is industrially applicable

Claims

THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS:

1. A production method of decursin, wherein the method takes Angelica decursiva as a raw material and comprises the steps of raw material crushing, extraction, refining and the like, and the extraction step comprises the step of carrying out ultrasonic extraction under the temperature of 40-70 °C and the frequency of 40-60 KHz by using a polar organic solvent as a solvent.

2. According to claim: the production method, wherein the polar organic solvent is selected from methanol, ethanol, acetone or a mixed solvent of which the water content is lower than 20%, and 90%-95% of ethanol is preferred selected.

3. The production method according to claims 1-2, wherein the refining step comprises: extracting an extract obtained by ultrasonic extraction with chloroform or a solvent which has a polarity similar to that of chloroform, drying the obtained extract at a low temperature, and carrying out column chromatography separation.

4. The production method according to claim 3, wherein the low-temperature drying is preferably a microwave drying method.

5. The production method according to claim 4, wherein the column chromatography is preferably silica gel column chromatography and gradient elution is carried out with n-hexane and ethyl acetate as an eluent.

6. The production method according to claim 1, wherein the method comprises crushing Angelica decursiva into 20-60 meshes of powder, carrying out ultrasonic extraction under the temperature of 40-70 °C and the frequency of 40-60 KHz for 30 min with 90-95% ethanol, carrying out extraction for twice, mixing extractive solutions, carrying out filtration, recovering ethanol from the filtrate, carrying out dilution with water, carrying out extraction with chloroform for 3-5 times, mixing polar layers, recovering chloroform, carrying out microwave drying at a low temperature, carrying out chromatography on a silica gel column, carrying out gradient elution with a mixed solvent of n-hexane and ethyl acetate, and collecting decursin.

7. The production method according to claims 1-6, wherein the method further comprises the step of further purifying the obtained decursin.

8. The production method according to claim 7, wherein the purification is column chromatography or recrystallization.