Method for extracting and separating artemisinin from artemisinin crystallization mother liquor

Abstract

The invention discloses a method for extracting and separating artemisinin alkene from artemisinin crystallization mother liquor, which comprises the following steps: step 1, separating an artemisinin crude product from an artemisinin extracting solution, crystallizing the artemisinin crude product, and concentrating the obtained crystallization mother liquor to constant weight; 2, extracting the concentrate with a solvent 1, and filtering; step 3, concentrating and drying the filtrate; 4, crystallizing the concentrate by using a solvent 2 to obtain an artemisinin crude product; 5, recrystallizing the crude artemisinin product with a solvent 2 to obtain a wet artemisinin product; and 6, drying the wet artemisinin product to obtain the artemisinin product. The invention provides a scheme for separating and purifying the artemisinin from the artemisia apiacea crystal mother liquor by a solvent method for the first time, the obtained artemisinin is high in content and yield, and the method disclosed by the invention is relatively simple in used equipment, simple in process, few in used solvent type, good in stability and repeatability, very good in popularization and application prospect, and suitable for industrial production. And the obtained artemisinene product can be applied to various fields such as cosmetics and medicines at the same time.

Description

Technical Field

[0001] The present invention relates to the technical field of artemisinin preparation, and in particular to a method for extracting and separating artemisinin from artemisinin crystallization mother liquor. Background Art

[0002] Artemisinin is a natural product extracted from Artemisia annua plants, and its structure is shown below:

[0003]

[0004] Artemisinin has been widely used in the treatment of malaria. Its pharmacological characteristics are that it selectively kills malarial parasites without affecting normal human cells. Studies in recent years have shown that artemisinin also has a certain selective killing effect on cells, so it has attracted widespread attention from scientists. The study of the selective killing effect of tumor cells and its mechanism of action has important implications for the discovery of the wide application of artemisinin and will also help discover and develop new anti-cancer drugs.

[0005] The market demand for artemisinin is growing day by day, and it is mainly used in cosmetics, medicine and other facial fields. However, there are few preparation processes for artemisinin in the existing technology, and it is necessary to enrich the preparation scheme of artemisinin. Summary of the invention

[0006] The technical problem to be solved by the present invention is to provide a method for extracting and separating artemisinin from artemisinin crystal mother liquor in view of the deficiencies in the above-mentioned prior art.

[0007] In order to solve the above technical problems, the technical solution adopted by the present invention is: a method for extracting and separating artemisinin from artemisinin crystal mother liquor, characterized in that it comprises the following steps:

[0008] Step 1: separating crude artemisinin from the artemisinin extract, crystallizing the crude artemisinin, and concentrating the resulting crystallization mother liquor to a constant weight;

[0009] Step 2: extracting the concentrate obtained in step 1 with solvent 1 and filtering;

[0010] Step 3: Concentrating the filtrate obtained in step 2 to obtain a concentrate;

[0011] Step 4: crystallizing the concentrate obtained in step 3 with solvent 2 to obtain a crude product of artemisinene;

[0012] Step 5: Recrystallize the crude artemisinin with solvent 2 to obtain wet artemisinin;

[0013] Step 6: Dry the wet artemisinin product to obtain the artemisinin product.

[0014] Preferably, the solvent 1 is at least one of methanol, ethanol and ethyl acetate.

[0015] Preferably, the solvent 2 is ethanol or a mixture of ethanol and water.

[0016] Preferably, the volume fraction of ethanol in the ethanol-water mixture is 30-80%.

[0017] Preferably, the concentration temperature in step 1 is 40-80°C, and the concentration temperature in step 3 is 60-90°C.

[0018] Preferably, the extraction temperature in step 2 is 50-90°C.

[0019] Preferably, the extraction times in step 2 are 1 to 3 times, and the extraction time is 1 to 3 hours.

[0020] Preferably, the crystallization temperature in step 4 and step 5 is 0°C to -10°C.

[0021] Preferably, the drying temperature in step six is ​​40-70°C.

[0022] Preferably, the method for extracting and separating artemisinin from artemisinin crystallization mother liquor comprises the following steps:

[0023] Step 1: separating crude artemisinin from the artemisinin extract, crystallizing the crude artemisinin, and concentrating the resulting crystallization mother liquor to a constant weight;

[0024] Step 2: extract the concentrate obtained in step 1 with methanol, extract twice, each time for 2 hours, extraction temperature 60°C, combine the extracts obtained from the two times, and filter;

[0025] Step 3: Concentrate the filtrate obtained in step 2 under vacuum at 60° C. to dryness to obtain a concentrate;

[0026] Step 4: crystallizing the concentrate obtained in step 3 with ethanol having a volume concentration of 80% at -5°C to obtain a crude product of artemisinene;

[0027] Step 5: recrystallizing the crude artemisinin with ethanol having a volume concentration of 80% at -5°C to obtain a wet artemisinin product;

[0028] Step six: Dry the wet artemisinin at 60°C to obtain the artemisinin product.

[0029] The beneficial effects of the present invention are:

[0030] The present invention provides a method for extracting and separating artemisinin from artemisinin crystal mother liquor. The present invention provides for the first time a scheme for separating and purifying artemisinin from artemisinin crystal mother liquor by a solvent method. The obtained artemisinin has a high content and a high yield. The method of the present invention has relatively simple equipment and process, uses fewer types of solvents, has good stability and repeatability, and has a good prospect for promotion and application. The obtained artemisinin product can also be applied to various fields such as cosmetics and medicines. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 The chromatograms of artemisinin prepared in Example 1 and the reference substance artemisinin;

[0032] Figure 2 This is the chromatogram of the reference substance artemisinin. DETAILED DESCRIPTION

[0033] The present invention is further described in detail below in conjunction with embodiments so that those skilled in the art can implement the invention with reference to the description.

[0034] It should be understood that the terms such as “having”, “including” and “comprising” used herein do not exclude the existence or addition of one or more other elements or combinations thereof.

[0035] The test methods used in the following examples are conventional methods unless otherwise specified. The materials and reagents used in the following examples are all commercially available unless otherwise specified. In the following examples, if no specific conditions are specified, the experiments were carried out under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used, if the manufacturer is not specified, are all conventional products that can be purchased commercially.

[0036] Example 1

[0037] A method for extracting and separating artemisinin from artemisinin crystallization mother liquor comprises the following steps:

[0038] Step 1: separating crude artemisinin from the artemisinin extract, crystallizing the crude artemisinin at 20° C., concentrating the crystallization mother liquor to a constant weight, and obtaining 10 kg of crystallization mother liquor concentrate;

[0039] Step 2: The concentrate was extracted with methanol twice, each time for 2 hours, the extraction multiple was 2 times (the mass unit was kg, the volume unit was L, the volume of methanol extracted each time was 2 times the mass of the concentrate, the same as in the following examples), the extraction temperature was 60° C., the extracts obtained from the two times were combined and filtered;

[0040] Step 3: Concentrate the filtrate obtained in step 2 under vacuum at 60°C to dryness to obtain 0.38 kg;

[0041] Step 4: Dissolve 0.38 kg of the concentrate in 80% by volume ethanol at 60° C., and then crystallize at -5° C. to obtain 0.26 kg of crude artemisinin;

[0042] Step 5: Dissolve 0.26 kg of crude artemisinin in 80% by volume ethanol at 60°C, and then crystallize at -5°C to obtain 0.23 kg of wet artemisinin;

[0043] Step six: Dry 0.23 kg of wet artemisinin obtained in step five at 60° C. to obtain 0.20 kg of artemisinin, which is the final product.

[0044] Example 2

[0045] A method for extracting and separating artemisinin from artemisinin crystallization mother liquor comprises the following steps:

[0046] Step 1: separating crude artemisinin from the artemisinin extract, crystallizing the crude artemisinin at 20° C., concentrating the crystallization mother liquor to a constant weight, and obtaining 50 kg of crystallization mother liquor concentrate;

[0047] Step 2: extract the concentrate with methanol twice, 2 hours each time, the extraction multiple is 2 times, the temperature is 60°C, combine the extracts obtained from the two times, and filter;

[0048] Step 3: Concentrate the filtrate obtained in step 2 under vacuum at 60°C and dry to obtain 1.98 kg;

[0049] Step 4: Dissolve 1.98 kg of the concentrate in 80% by volume ethanol at 60°C, and crystallize at -5°C to obtain 1.32 kg of crude artemisinin;

[0050] Step 5: Dissolve 1.32 kg of crude artemisinin in 80% by volume ethanol at 60°C, and then crystallize at -5°C to obtain 1.21 kg of wet artemisinin;

[0051] Step 6: Dry 1.21 kg of wet artemisinin obtained in step 5 at 60° C. to obtain 1.02 kg of artemisinin, which is the final product.

[0052] Example 3

[0053] A method for extracting and separating artemisinin from artemisinin crystallization mother liquor comprises the following steps:

[0054] Step 1: separating crude artemisinin from the artemisinin extract, crystallizing the crude artemisinin at 20° C., concentrating the crystallization mother liquor to a constant weight, and obtaining 500 kg of crystallization mother liquor concentrate;

[0055] Step 2: extract the concentrate with methanol twice, 2 hours each time, the extraction multiple is 2 times, the temperature is 60°C, combine the extracts obtained from the two times, and filter;

[0056] Step 3: Concentrate the filtrate obtained in step 2 under vacuum at 60°C to dryness to obtain 19.5 kg;

[0057] Step 4: Dissolve 19.5 kg of the concentrate in 80% by volume ethanol at 60°C, and crystallize at -5°C to obtain 13.2 kg of crude artemisinin;

[0058] Step 5: Dissolve 13.2 kg of crude artemisinin in 80% by volume ethanol at 60°C, and then crystallize at -5°C to obtain 11.7 kg of wet artemisinin;

[0059] Step 6: Dry 11.7 kg of wet artemisinin obtained in step 5 at 60° C. to obtain 10.2 kg of artemisinin, which is the final product.

[0060] Product Testing

[0061] Determined according to high performance liquid chromatography (General Rule 0512);

[0062] Chromatographic conditions and system suitability test:

[0063] Column: Diamonsil C18 5μm 250x 4.6mm

[0064] Chromatographic conditions:

[0065] Mobile phase: methanol-water (volume ratio 75:25)

[0066] Flow rate: 1ml / min

[0067] Injection volume: 20 μL

[0068] Detector: Evaporative light scattering detection

[0069] The number of theoretical plates calculated based on peaks should not be less than 2000.

[0070] Preparation of reference solution: An appropriate amount of artemisinin reference substance (commercially available artemisinin product, purchased from Changzhou Chenhong Biotechnology Co., Ltd., batch number 20231212) was accurately weighed and methanol was added to make a solution containing 0.1 mg per 1 ml.

[0071] Preparation of test solution: Take 2 parts of each of artemisinene prepared in Examples 1 to 3 above (precisely weighed, placed in a stoppered conical flask, accurately add 10 ml of methanol, weigh the weight, ultrasonically treat (power 250 W, frequency 25 kHz) for 15 minutes, cool, weigh again, make up the lost weight with methanol, shake well, filter, and obtain;

[0072] Determination method: Accurately pipette 10 μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0073] The chromatograms of artemisinin prepared in Example 1 and the reference artemisinin are as follows: Figure 1 and Figure 2 shown.

[0074] The content detection results and the artemisinin yield results are shown in Table 1 below:

[0075] Table 1

[0076]

[0077]

[0078] According to the sample content and yield results of the above three examples, it is shown that the determination results are relatively stable, the artemisinin products of Examples 1-3 have high purity, high artemisinin product yield, high process stability and good repeatability.

[0079] Although the embodiments of the present invention have been disclosed as above, they are not limited to the applications listed in the specification and the implementation modes. They can be fully applied to various fields suitable for the present invention. For those familiar with the art, additional modifications can be easily implemented. Therefore, without departing from the general concept defined by the claims and the scope of equivalents, the present invention is not limited to specific details.

Claims

1. A method for extracting and separating artemisinin from artemisinin crystal mother liquor, characterized in that: The following steps are involved: Step 1: separating crude artemisinin from the artemisinin extract, crystallizing the crude artemisinin, and concentrating the resulting crystallization mother liquor to a constant weight; Step 2: extracting the concentrate obtained in step 1 with solvent 1 and filtering; Step 3: Concentrating the filtrate obtained in step 2 to obtain a concentrate; Step 4: crystallizing the concentrate obtained in step 3 with solvent 2 to obtain a crude product of artemisinene; Step 5: Recrystallize the crude artemisinin with solvent 2 to obtain wet artemisinin; Step 6: Dry the wet artemisinin product to obtain the artemisinin product.

2. The method for extracting and separating artemisinin from artemisinin crystal mother liquor according to claim 1, characterized in that: The solvent 1 is at least one of methanol, ethanol and ethyl acetate.

3. The method for extracting and separating artemisinin from artemisinin crystal mother liquor according to claim 1, characterized in that: Solvent 2 is ethanol or a mixture of ethanol and water.

4. The method for extracting and separating artemisinin from artemisinin crystal mother liquor according to claim 3, characterized in that: The volume fraction of ethanol in the ethanol-water mixture is 30-80%.

5. The method for extracting and separating artemisinin from artemisinin crystal mother liquor according to claim 1, characterized in that: The concentration temperature in step 1 is 40-80°C, and the concentration temperature in step 3 is 60-90°C.

6. The method for extracting and separating artemisinin from artemisinin crystal mother liquor according to claim 1, characterized in that: The extraction temperature in step 2 is 50-90°C.

7. The method for extracting and separating artemisinin from artemisinin crystal mother liquor according to claim 6, characterized in that: The extraction times in step 2 are 1 to 3 times, and the extraction time is 1 to 3 hours.

8. The method for extracting and separating artemisinene from artemisinin crystal mother liquor according to claim 1, characterized in that: The crystallization temperature in step 4 and step 5 is 0°C to -10°C.

9. The method for extracting and separating artemisinene from artemisinin crystal mother liquor according to claim 1, characterized in that: In step six, the drying temperature is 40-70°C.

10. The method for extracting and separating artemisinene from artemisinin crystal mother liquor according to any one of claims 1 to 9, characterized in that: The following steps are involved: Step 1: separating crude artemisinin from the artemisinin extract, crystallizing the crude artemisinin, and concentrating the resulting crystallization mother liquor to a constant weight; Step 2: extract the concentrate obtained in step 1 with methanol, extract twice, each time for 2 hours, extraction temperature 60°C, combine the extracts obtained from the two times, and filter; Step 3: Concentrate the filtrate obtained in step 2 under vacuum at 60° C. to dryness to obtain a concentrate; Step 4: crystallizing the concentrate obtained in step 3 with ethanol having a volume concentration of 80% at -5°C to obtain a crude product of artemisinene; Step 5: recrystallizing the crude artemisinin with ethanol having a volume concentration of 80% at -5°C to obtain a wet artemisinin product; Step six: Dry the wet artemisinin at 60°C to obtain the artemisinin product.

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