Method for extracting galanthamine from snow lotus
By using a specific ratio of solvent system and separation steps, high-purity galantamine was extracted from snow lotus, solving the problems of low purity and yield in existing technologies and achieving efficient galantamine separation.
Patent Information
- Application Number
- CN202511741534.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-11-25
- Publication Date
- 2026-02-10
AI Technical Summary
The purity and yield of galantamine extracted from Amaryllidaceae plants in the existing technology are low.
Snowdrop was extracted at 60-90℃ using a mixed solvent of ethyl acetate, methanol, and acetone in a specific ratio. The extraction was combined with alkaline aqueous solution extraction, silica gel column separation, and elution with petroleum ether and methanol in a specific ratio. Finally, high-purity galantamine was obtained by crystallization.
It significantly improves the yield and purity of galantamine, is simple to operate, and has excellent separation effect.
Smart Images
Figure CN121494864A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of natural product extraction technology, specifically relating to a method for extracting galantamine from snow lotus. Background Technology Galanthamine is a phenanthridine alkaloid extracted from plants in the Amaryllidaceae family (such as snowdrop and Lycoris radiata), and can also be chemically synthesized. It belongs to the reversible acetylcholinesterase inhibitor class and is clinically used to treat mild to moderate Alzheimer's disease (senile dementia), and also as adjunctive therapy for neurological disorders such as myasthenia gravis and sequelae of poliomyelitis. It is a prescription drug, belonging to the second-generation cholinesterase inhibitor class, and has a dual mechanism of action: it not only inhibits acetylcholinesterase but also regulates the function of nicotinic acetylcholine receptors (nAChR).
[0002] The global galantamine market has shown a growth trend in recent years, especially in oral and injectable formulations. Market demand continues to grow, and the increasing global aging trend and rising diagnosis rates of cognitive impairment diseases such as Alzheimer's disease are expected to drive steady growth in demand for therapeutic drugs such as galantamine.
[0003] In existing technologies, the extraction of galantamine from Amaryllidaceae plants suffers from low purity and low yield. Summary of the Invention
[0004] The technical problem to be solved by the present invention is to overcome the deficiencies and defects mentioned in the background art above, and to provide a method for extracting galantamine from snow lotus in order to improve the yield and purity.
[0005] To solve the above-mentioned technical problems, the technical solution proposed by this invention is as follows: A method for extracting galantamine from snowdrop includes the following steps: S1. Wash, crush, and dry the snowflakes; S2. The dried snowflake is extracted with an extraction solvent to obtain an extract, and the extract is concentrated to obtain a crude extract; the extraction solvent is a mixed solvent of ethyl acetate, methanol and acetone in a volume ratio of 1:(0.85~1.15):(0.15~0.25); the extraction temperature is 60~90℃; S3. Add the crude extract to an alkaline aqueous solution, stir and separate the layers, then extract the alkaline aqueous layer with an extractant, and concentrate the extract to obtain crude galantamine. S4. The crude galantamine is subjected to column separation and then concentrated to obtain high-content galantamine; the column separation is carried out using a silica gel column, and the elution solvent is petroleum ether and methanol in a volume ratio of 5~20:1. S5. Crystallize the high-content galantamine to obtain galantamine.
[0006] As a further improvement, the extraction solvent described in S2 is a mixture of ethyl acetate, methanol, and acetone in a volume ratio of 1:1:0.2.
[0007] As a further improvement, the extraction temperature described in S2 is 80°C.
[0008] As a further improvement, the extraction time described in S2 is 1 to 3 hours, and the number of extractions is 1 to 3 times.
[0009] As a further improvement, the aqueous solution of the alkali in S3 is at least one of 1-5 wt% sodium hydroxide, saturated sodium bicarbonate, and 10-15 wt% potassium hydroxide.
[0010] As a further improvement, the extractant in S3 is at least one of petroleum ether, ethyl acetate, acetone, and chloroform.
[0011] As a further improvement, the weight ratio of silica gel to sample loading in the silica gel column described in S4 is 5~12:1.
[0012] As a further improvement, the elution solvent in S4 is petroleum ether and methanol in a volume ratio of 6:1.
[0013] As a further improvement, the solvent for crystallization in S5 is at least one of petroleum ether, methanol, acetone, and ethanol.
[0014] As a further improvement, the crystallization temperature described in S5 is -10 to 10°C.
[0015] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention uses a mixed solvent of ethyl acetate, methanol, and acetone in a specific ratio with different polarities as the extraction solvent, and at an extraction temperature of 60-90°C, galantamine from snowdrop can be extracted into the solution to the maximum extent, while other interfering substances and impurities are extracted as little as possible. Using a specific ratio of petroleum ether and methanol as the elution solvent can better separate galantamine from impurities, thereby improving the yield and purity of galantamine.
[0016] The method provided by this invention is simple to operate, has excellent extraction and separation effects, and achieves high yield and purity of galantamine. Attached Figure Description
[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.
[0018] Figure 1 This is the chromatogram of galantamine reference standard; Figure 2 This is the chromatogram of the galantamine sample obtained in Example 1. Detailed Implementation
[0019] To facilitate understanding of the present invention, the present invention will be described more fully and in detail below with reference to the accompanying drawings and preferred embodiments, but the scope of protection of the present invention is not limited to the following specific embodiments.
[0020] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by those skilled in the art. The technical terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the scope of the invention.
[0021] Unless otherwise specified, all raw materials, reagents, instruments and equipment used in this invention can be purchased from the market or prepared by existing methods.
[0022] In some specific embodiments, the method for extracting galantamine from snowdrop according to the present invention includes the following steps: S1. Wash, crush, and dry the snowflakes.
[0023] S2. The dried snowflakes were extracted with an extraction solvent to obtain an extract, and the extract was concentrated to obtain a crude extract.
[0024] In some embodiments, the extraction solvent is a mixture of ethyl acetate, methanol, and acetone in a volume ratio of 1:(0.85~1.15):(0.15~0.25) (preferably 1:1:0.2). Preferably, the volume ratio of the extraction solvent to the mass of the dried snowflake is 10:1 (denoted as 10V).
[0025] In some embodiments, the extraction temperature is 60~90℃, preferably 80℃.
[0026] In some embodiments, the extraction time is 1 to 3 hours, the extraction is performed 1 to 3 times, and the extracts are combined.
[0027] S3. Add the crude extract to an alkaline aqueous solution, stir and separate the layers to remove impurities through alkalization, then extract the alkaline aqueous layer with an extractant, and concentrate the extract to obtain crude galantamine.
[0028] In some embodiments, the aqueous solution of the alkali is at least one selected from 1-5 wt% sodium hydroxide, saturated sodium bicarbonate, and 10-15 wt% potassium hydroxide. Preferably, the volume ratio of the aqueous solution of the alkali to the mass of the crude extract is 5:1 (denoted as 5V).
[0029] In some embodiments, the extractant is at least one selected from petroleum ether, ethyl acetate, acetone, and chloroform.
[0030] S4. The crude galantamine is separated by column chromatography and concentrated to obtain high-content galantamine.
[0031] In some embodiments, column separation is performed using a silica gel column with a mesh size of 100-300 mesh, and the ratio of silica gel to sample loading is 5-12:1 by weight.
[0032] In some embodiments, the elution solvent for column separation is petroleum ether and methanol in a volume ratio of 5 to 20:1 (preferably 6:1).
[0033] S5. Crystallize the high-content galantamine to obtain galantamine.
[0034] In some embodiments, the crystallization solvent is at least one of petroleum ether, methanol, acetone, and ethanol, preferably 50% (by volume) ethanol.
[0035] In some embodiments, the crystallization temperature is -10 to 10°C.
[0036] Example 1 1. Wash, crush, and dry the snowflakes; 2. Weigh 100g of snow lotus medicinal material, and extract twice with 10V ethyl acetate, methanol and acetone (1:1:0.2 volume) at 80℃ for 2 hours each time. Combine the two extracts and concentrate the extract at 70℃ to obtain 16.5g of crude extract. 3. 16.5g of crude extract was stirred with 5V 2% sodium hydroxide alkaline solution, and the layers were separated. The alkaline layer was then extracted with 2V chloroform. The chloroform extract was concentrated at 60℃ to obtain 6.2g of crude galantamine. 4. Mix 6.2g of crude galantamine with 6.2g of 200-300 mesh silica gel, then pack the sample into a column with 62g of 200-300 mesh silica gel. Elute with petroleum ether:methanol 6:1 and collect the solution containing galantamine as the main component. Concentrate the solution to obtain 0.98g of high-content galantamine. 5. Dissolve 0.98g of high-content galantamine in 50% ethanol by heating at 75°C, crystallize at 5°C, filter, and obtain 0.43g of galantamine crystals.
[0037] Example 2 1. Wash, crush, and dry the snowflakes; 2. Weigh 500g of snow lotus medicinal material, and extract twice with 10V ethyl acetate, methanol and acetone (1:1:0.2 volume) at 80℃ for 2 hours each time. Combine the two extracts and concentrate the extract at 70℃ to obtain 81.6g of crude extract. 3. 81.6g of crude extract was stirred with 5V 2% sodium hydroxide alkaline solution, and the layers were separated. The alkaline layer was then extracted with 2V chloroform. The chloroform extract was concentrated at 60℃ to obtain 30.5g of crude galantamine. 4. Mix 30.5g of crude galantamine with 30.5g of 200-300 mesh silica gel, then pack the sample into a column with 305g of 200-300 mesh silica gel. Elute with petroleum ether:methanol 6:1 and collect the sample. Collect the sample containing galantamine as the main component. Concentrate the sample to obtain 4.9g of high-content galantamine. 5. Dissolve 4.9g of high-content galantamine in 50% ethanol by heating at 75°C, crystallize at 5°C, filter, and obtain 2.13g of galantamine crystals.
[0038] Example 3 1. Wash, crush, and dry the snowflakes; 2. Weigh 10 kg of snow lotus medicinal material, and extract twice with 10 V ethyl acetate, methanol and acetone (1:1:0.2 volume) at 80℃ for 2 hours each time. Combine the two extracts and concentrate the extract at 70℃ to obtain 1.61 kg of crude extract. 3. 1.61 kg of crude extract was stirred with 5V 2% sodium hydroxide alkaline solution, and the layers were separated. The alkaline layer was then extracted with 2V chloroform. The chloroform extract was concentrated at 60℃ to obtain 0.60 kg of crude galantamine. 4. Mix 0.6 kg of crude galantamine with 0.6 kg of 200-300 mesh silica gel, then pack the sample into a column with 6 kg of 200-300 mesh silica gel. Elute with petroleum ether:methanol 6:1 and collect the sample. Collect the liquid containing galantamine as the main component. Concentrate the liquid to obtain 95.2 g of high-content galantamine. 5. Dissolve 95.2g of high-content galantamine in 50% ethanol by heating at 75°C, crystallize at 5°C, filter, and obtain 43.5g of galantamine crystals.
[0039] Comparative Example 1 1. Wash, crush, and dry the snowflakes; 2. Weigh 500g of snow lotus medicinal material, and extract twice with 10V ethyl acetate, methanol and acetone (1:1:1 volume) at 80℃ for 2 hours each time. Combine the two extracts and concentrate the extract at 70℃ to obtain 61.6g of crude extract. 3. 61.6g of crude extract was stirred with 5V 2% sodium hydroxide alkaline solution, and the layers were separated. The alkaline layer was then extracted with 2V chloroform. The chloroform extract was concentrated at 60℃ to obtain 19.5g of crude galantamine. S4. Mix 19.5g of crude galantamine with 19.5g of 200-300 mesh silica gel, then pack the sample into a column with 195g of 200-300 mesh silica gel. Elute with petroleum ether:methanol 6:1 and collect the sample. Collect the sample containing galantamine as the main component. Concentrate the sample to obtain 1.7g of high-content galantamine. S5. Dissolve 1.7g of high-content galantamine in 50% ethanol by heating at 75°C, crystallize at 5°C, filter, and obtain 0.95g of galantamine crystals.
[0040] Comparative Example 2 1. Wash, crush, and dry the snowflakes; 2. Weigh 500g of snow lotus medicinal material, and extract twice with 10V ethyl acetate, methanol and acetone (0.2:1:1 volume) at 80℃ for 2 hours each time. Combine the two extracts and concentrate the extract at 70℃ to obtain 80.5g of crude extract. 3. 80.5g of crude extract was stirred with 5V 2% sodium hydroxide alkaline solution, and the layers were separated. The alkaline layer was then extracted with 2V chloroform. The chloroform extract was concentrated at 60℃ to obtain 31.2g of crude galantamine. S4. Mix 31.2g of crude galantamine with 31.2g of 200-300 mesh silica gel, then pack the sample into a column with 312g of 200-300 mesh silica gel. Elute with petroleum ether:methanol 6:1 and collect the sample. Collect the sample containing galantamine as the main component. Concentrate the sample to obtain 1.92g of high-content galantamine. S5. Dissolve 1.92g of high-content galantamine in 50% ethanol by heating at 75°C, crystallize at 5°C, filter, and obtain 1.95g of galantamine crystals.
[0041] Comparative Example 3 1. Wash, crush, and dry the snowflakes; 2. Weigh 500g of snow lotus medicinal material, and extract twice at 80℃ with 10V ethyl acetate, methanol and acetone (volume ratio 1:0.2:1), each time for 2 hours. Combine the two extracts and concentrate the extract at 70℃ to obtain 70.5g of crude extract. 3. 70.5g of crude extract was stirred with 5V 2% sodium hydroxide alkaline solution, and the layers were separated. The alkaline layer was then extracted with 2V chloroform. The chloroform extract was concentrated at 60℃ to obtain 24.2g of crude galantamine. S4. Mix 24.2g of crude galantamine with 24.2.2g of 200-300 mesh silica gel, then pack the sample into a column with 242g of 200-300 mesh silica gel. Elute with petroleum ether:methanol 6:1 and collect the solution containing galantamine as the main component. Concentrate the solution to obtain 1.12g of high-content galantamine. S5. Dissolve 1.12g of high-content galantamine in 50% ethanol by heating at 75°C, crystallize at 5°C, filter, and obtain 0.95g of galantamine crystals.
[0042] Comparative Example 4 1. Wash, crush, and dry the snowflakes; 2. Weigh 500g of snow lotus medicinal material, and extract twice with 10V ethyl acetate, ethanol and acetone (1:1:0.2 volume) at 80℃ for 2 hours each time. Combine the two extracts and concentrate the extract at 70℃ to obtain 48.6g of crude extract. 3. 48.6g of crude extract was stirred with 5V 2% sodium hydroxide alkaline solution, and the layers were separated. The alkaline layer was then extracted with 2V chloroform. The chloroform extract was concentrated at 60℃ to obtain 11.5g of crude galantamine. S4. Mix 11.5g of crude galantamine with 11.5g of 200-300 mesh silica gel, then pack the sample into a column with 195g of 200-300 mesh silica gel. Elute with petroleum ether:methanol 6:1 and collect the sample. Collect the sample containing galantamine as the main component. Concentrate the sample to obtain 1.2g of high-content galantamine. S5. Dissolve 1.2g of high-content galantamine in 50% ethanol by heating at 75°C, crystallize at 5°C, filter, and obtain 0.65g of galantamine crystals.
[0043] Comparative Example 5 1. Wash, crush, and dry the snowflakes; 2. Weigh 500g of snow lotus medicinal material, and extract twice at 80℃ with 10V petroleum ether, methanol and acetone (volume ratio 1:1:0.2), each time for 2 hours. Combine the two extracts and concentrate the extract at 70℃ to obtain 51.4g of crude extract. 3. 51.4g of crude extract was stirred with 5V 2% sodium hydroxide alkaline solution, and the layers were separated. The alkaline layer was then extracted with 2V chloroform. The chloroform extract was concentrated at 60℃ to obtain 14.5g of crude galantamine. S4. Mix 14.5g of crude galantamine with 14.5g of 200-300 mesh silica gel, then pack the sample into a column with 195g of 200-300 mesh silica gel. Elute with petroleum ether:methanol 6:1 and collect the sample. Collect the sample containing galantamine as the main component. Concentrate the sample to obtain 1.1g of high-content galantamine. S5. Dissolve 1.1g of high-content galantamine in 50% ethanol by heating at 75°C, crystallize at 5°C, filter, and obtain 0.55g of galantamine crystals.
[0044] Comparative Example 6 1. Wash, crush, and dry the snowflakes; 2. Weigh 500g of snow lotus medicinal material, and extract twice with 10V ethyl acetate, methanol and acetone (volume ratio 1:1:0.2) at 80℃ for 2 hours each time. Combine the two extracts and concentrate the extract at 70℃ to obtain 61.3g of crude extract. 3. 61.3g of crude extract was stirred with 5V 2% sodium hydroxide alkaline solution, and the layers were separated. The alkaline layer was then extracted with 2V chloroform. The chloroform extract was concentrated at 60℃ to obtain 19.1g of crude galantamine. S4. Mix 19.1g of crude galantamine with 19.1g of 200-300 mesh silica gel, then pack the sample into a column with 195g of 200-300 mesh silica gel. Elute with petroleum ether:methanol 4:1 and collect the sample. Collect the sample containing galantamine as the main component. Concentrate the sample to obtain 1.4g of high-content galantamine. S5. Dissolve 1.4g of high-content galantamine in 50% ethanol by heating at 75°C, crystallize at 5°C, filter, and obtain 0.90g of galantamine crystals.
[0045] Product Testing Determined according to high performance liquid chromatography (General Rule 0512); Chromatographic conditions and system suitability test: Column: C185μm 150 x 4.6mm Chromatographic conditions: Mobile phase: Acetonitrile / water (volume ratio 55:45) Flow rate: 1 ml / min Injection volume: 10 μL Column temperature: 25℃ The theoretical plate number, calculated based on the peak, should be no less than 2000.
[0046] Preparation of reference solution: Accurately weigh an appropriate amount of galantamine reference standard, add methanol to prepare a solution containing 0.1 mg per ml; Preparation of test solution: Take two portions of galantamine obtained from the above examples and comparative examples (accurately weighed, placed in a stoppered conical flask, accurately add 10 ml of methanol, weigh, sonicate (power 250 W, frequency 25 kHz) for 15 minutes, cool, weigh again, make up the lost weight with methanol, shake well, filter, and the test solution is obtained. Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.
[0047] Figure 1 This is the chromatogram of galantamine reference standard; Figure 2 This is the chromatogram of the galantamine sample obtained in Example 1.
[0048] The results of galantamine content detection and galantamine yield for each example and comparative example are shown in Table 1 below (wherein, the HPLC content was calculated by peak area normalization method, and the yield of the refined product in Example 2 was 94.67%, based on the total amount of galantamine in the medicinal material): Table 1
[0049] Based on the above examples and comparative sample content and yield results, it is shown that the galantamine products of Examples 1-3 have high purity, high yield, high process stability, and good repeatability.
[0050] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention in any way. Therefore, any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention should fall within the protection scope of the present invention.
Claims
1. A method for extracting galantamine from snowdrop, characterized in that, Includes the following steps: S1. Wash, crush, and dry the snowflakes; S2. The dried snowflake is extracted with an extraction solvent to obtain an extract, and the extract is concentrated to obtain a crude extract; the extraction solvent is a mixed solvent of ethyl acetate, methanol and acetone in a volume ratio of 1:(0.85~1.15):(0.15~0.25); the extraction temperature is 60~90℃; S3. Add the crude extract to an alkaline aqueous solution, stir and separate the layers, then extract the alkaline aqueous layer with an extractant, and concentrate the extract to obtain crude galantamine. S4. The crude galantamine is subjected to column separation and then concentrated to obtain high-content galantamine; the column separation is carried out using a silica gel column, and the elution solvent is petroleum ether and methanol in a volume ratio of 5~20:
1. S5. Crystallize the high-content galantamine to obtain galantamine.
2. The method for extracting galantamine from snowdrop according to claim 1, characterized in that, The extraction solvent described in S2 is a mixture of ethyl acetate, methanol, and acetone in a volume ratio of 1:1:0.
2.
3. The method for extracting galantamine from snowdrop according to claim 1 or 2, characterized in that, The extraction temperature described in S2 is 80℃.
4. The method for extracting galantamine from snowdrop according to claim 1 or 2, characterized in that, The extraction time described in S2 is 1 to 3 hours, and the number of extractions is 1 to 3 times.
5. The method for extracting galantamine from snowdrop according to claim 1, characterized in that, The aqueous solution of the alkali mentioned in S3 is at least one of 1-5 wt% sodium hydroxide, saturated sodium bicarbonate, and 10-15 wt% potassium hydroxide.
6. The method for extracting galantamine from snowdrop according to claim 1 or 5, characterized in that, The extractant mentioned in S3 is at least one of petroleum ether, ethyl acetate, acetone, and chloroform.
7. The method for extracting galantamine from snowdrop according to claim 1, characterized in that, The weight ratio of silica gel used in the silica gel column to the amount of sample loaded in S4 is 5~12:
1.
8. The method for extracting galantamine from snowdrop according to claim 1 or 7, characterized in that, The elution solvent in S4 is petroleum ether and methanol in a volume ratio of 6:
1.
9. The method for extracting galantamine from snowdrop according to claim 1, characterized in that, The solvent for crystallization described in S5 is at least one of petroleum ether, methanol, acetone, and ethanol.
10. The method for extracting galantamine from snowdrop according to claim 1 or 9, characterized in that, The crystallization temperature described in S5 is -10 to 10°C.