CN110746417A - Extraction method of berberine - Google Patents
Extraction method of berberine Download PDFInfo
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- C07D455/00—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine
- C07D455/03—Heterocyclic compounds containing quinolizine ring systems, e.g. emetine alkaloids, protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine containing quinolizine ring systems directly condensed with at least one six-membered carbocyclic ring, e.g. protoberberine; Alkylenedioxy derivatives of dibenzo [a, g] quinolizines, e.g. berberine
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Abstract
The invention relates to a method for extracting berberine, and belongs to the technical field of extraction of active ingredients of medicinal plants. The extraction method of the berberine comprises the following steps: putting the traditional Chinese medicine powder and an extraction solvent into an extraction container, sealing the extraction container, extracting at 100-140 ℃, and performing solid-liquid separation after extraction to obtain a berberine extracting solution; the extraction solvent is a mixed solution of ethanol and water, and the volume concentration of the ethanol in the extraction solvent is 30-70%; the Chinese medicinal powder is selected from one or two of Coptidis rhizoma powder and cortex Phellodendri powder. The extraction method of berberine of the invention has simple operation, no volatilization of extraction solvent, safety and innocuity, cheap and easily obtained required equipment and low extraction cost; under the synergistic action of higher temperature and autogenous pressure, the method promotes the cracking or hydrolysis of plant cell walls, so that active components in cells can be efficiently and quickly dissolved out, and the extraction rate and the extraction efficiency of berberine are effectively improved.
Description
Technical Field
The invention relates to a method for extracting berberine, and belongs to the technical field of extraction of active ingredients of medicinal plants.
Background
The Coptidis rhizoma is dried rhizome of Coptis chinensis Franch (Coptis chinensis Franch) belonging to Ranunculaceae. Coptis root, rhizoma Coptidis is cold in nature and bitter in taste, and has effects of clearing heat, eliminating dampness, purging pathogenic fire and removing toxic substance, and it is used for treating diseases of heart, spleen, stomach, liver, gallbladder and large intestine. The main pharmacodynamic components of the coptis are alkaloids which mainly comprise berberine, palmatine, tetrandrine, berberrubine and the like, wherein the berberine is the alkaloid with the highest content in the coptis and is also the most main active substance of the coptis and has the effects of resisting diarrhea, resisting inflammation, promoting immunity, reducing blood pressure, resisting arrhythmia, reducing blood sugar and the like.
At present, various methods for extracting berberine from coptis chinensis are available, such as a reflux method, an extraction method, an ultrasonic-assisted method, a microwave method, a cellulase-assisted method and the like, wherein the ultrasonic-assisted extraction method and the microwave method need expensive extraction equipment, and the cellulase-assisted method needs enzymes with biological activity and has higher production cost. Therefore, the extraction methods of berberine are reflux method and extraction method. However, the two methods have long extraction time and low extraction rate when extracting berberine, and have certain limitation when being applied. For example, Zhang Xiaomei et al, in the research on the preparation process of Coptidis rhizoma extract based on different separation techniques (in the fourth 6 th period of 22 at the end of 3 months in 2016 (Chinese medicine guide), adopts ethanol reflux method to extract Coptidis rhizoma, and the specific method is as follows: weighing 2kg of coptis medicinal material, adding 40% ethanol (9 times, 5 times and 4 times) for reflux extraction for 3 times, extracting 1 time for 1.5h, extracting 2 times and 3 times for 45min respectively, combining 3 times of ethanol extract, concentrating to 7700mL (namely each 1mL is equivalent to 0.26g of raw material), taking 250mL of sample, freeze-drying (-50 ℃, 0.01-0.02 Mpas and 30h), and grinding. When the method is adopted to extract berberine from coptis, the extraction rate of berberine can only reach 4.7%.
Disclosure of Invention
The invention aims to provide an extraction method of berberine, which can improve the extraction rate of berberine.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
a method for extracting berberine comprises the following steps: putting the traditional Chinese medicine powder and an extraction solvent into an extraction container, sealing the extraction container, extracting at 100-140 ℃, and performing solid-liquid separation after extraction to obtain a berberine extracting solution;
the extraction solvent is a mixed solution of ethanol and water, and the volume concentration of the ethanol in the extraction solvent is 30-70%; the Chinese medicinal powder is selected from one or two of Coptidis rhizoma powder and cortex Phellodendri powder.
The invention adopts the solvothermal method to extract the berberine in the traditional Chinese medicine, and the physicochemical properties of the extraction solvent, such as polarity, dissolving capacity, dielectric constant, surface tension and the like, can generate mutation under the condition of the solvothermal method, thereby having the characteristics of high activity and high mass transfer rate. The extraction method of berberine of the invention has the following advantages: 1) the whole extraction process is carried out in a closed extraction system, so that the volatilization of an extraction solvent is effectively prevented; 2) the extraction solvent used in the extraction process is safe and nontoxic, and is beneficial to the post-treatment of the extract; 3) the extraction method is simple and convenient to operate, the required equipment is cheap and easy to obtain, and the extraction cost is low; 4) the solvent heat is extracted at a higher temperature, so that the extraction solvent can generate certain autogenous pressure in a closed extraction container, the cracking or hydrolysis of plant cell walls is promoted under the synergistic effect of the higher temperature and the autogenous pressure, the resistance of the solvent to permeate and diffuse into cells is reduced, active components in the cells can be efficiently and quickly dissolved out, the extraction efficiency and the extraction rate of the berberine are effectively improved, and the content of the berberine in the crude product is effectively improved, wherein the extraction rate of the berberine in the coptis chinensis is 6.01-8.53%, and the content of the berberine in the solid crude extract of the berberine extracted from the berberine in the coptis chinensis is 35-43%. In conclusion, the extraction method of berberine of the invention has the advantages of rapidness, high efficiency, environmental protection and the like.
The extraction time of the traditional Chinese medicine powder is not limited, but when the extraction time is too short, the extraction rate of the berberine is low, the traditional Chinese medicine is seriously wasted, and when the extraction time is too long, the extraction efficiency is reduced. Preferably, the extraction time is 10-60 min at 100-140 ℃.
In order to sufficiently dissolve berberine and reduce the dissolution of impurities, the volume of the extraction solvent used per 1g of the Chinese medicinal powder is preferably 10-60 mL.
Preferably, the traditional Chinese medicine powder is obtained by drying traditional Chinese medicines and then crushing the dried traditional Chinese medicines; the drying treatment temperature is 50-80 ℃, and the drying treatment time is 2-6 h.
Preferably, the particle size of the traditional Chinese medicine powder is 40-80 meshes. If the particle size is less than 40 meshes, the extraction solvent cannot completely permeate into the powder, so that incomplete extraction is caused, and the extraction rate is reduced; the particle size is higher than 80 meshes, the extraction rate of the berberine is not obviously different, but the centrifugal separation time needs to be prolonged. Therefore, the particle size of the preferred traditional Chinese medicine powder is 40-80 meshes.
Preferably, the filling rate of the traditional Chinese medicine powder and the extraction solvent in the extraction container is 60-80%. The influence of the filling rate on the extraction effect is shown as that the filling rate is lower than 60%, the autogenous pressure in the reaction kettle is low, the rupture of cell walls is not facilitated, and the extraction rate is low; if the filling rate is higher than 80%, although the extraction rate is high, the pressure in the reaction kettle is too high, and certain potential safety hazard exists.
Preferably, the concentration of ethanol in the extraction solvent is 30-70%. The higher the ethanol concentration in the extraction solvent, the higher the autogenous pressure in the reaction vessel under the same extraction conditions. However, if the concentration of ethanol in the extraction solvent is lower than 30%, the pressure in the reaction kettle is low, the damage degree to the cell wall of the powder is low, and the dissolution of active substances in plant cells is not facilitated; if the solubility of ethanol in the extraction solvent is more than 70%, the polarity of the extraction solvent is reduced, and berberine is a medium-polarity substance, and the polarity is too low to dissolve berberine according to the principle of similar compatibility. Therefore, the preferable ethanol concentration is 30 to 70%.
The extraction method of berberine can also be used for drying the obtained berberine extract to obtain crude extract of berberine. In addition, the obtained berberine extract can be further processed by other treatments, such as salting out to purify berberine.
Drawings
FIG. 1 is a standard curve used in the measurement of the extraction rate of berberine in experimental example 1.
Detailed Description
The present invention will be further described with reference to the following embodiments.
The volumes of the stainless steel reaction vessels used in the following examples were all 100 mL.
Example 1
The extraction method of berberine of the embodiment comprises the following steps:
1) drying Coptidis rhizoma at 50 deg.C for 6 hr, and pulverizing to 80 mesh to obtain Coptidis rhizoma powder;
2) putting the coptis powder obtained in the step 1) and the extraction solvent into a stainless steel reaction kettle with a polytetrafluoroethylene lining according to the proportion of 10mL of extraction solvent to each 1g of coptis powder, and controlling the filling rate to be 80%; the adopted extraction solvent is a mixed solution of ethanol and water, and the volume concentration of the ethanol in the mixed solution is 30 percent;
3) sealing the reaction kettle, putting the reaction kettle in an oven with the temperature of 100 ℃, and extracting for 10 minutes;
4) and after extraction is finished, taking out the reaction kettle from the oven, naturally cooling to room temperature, opening the reaction kettle, and centrifugally separating the content in the polytetrafluoroethylene reaction kettle to obtain the berberine extracting solution.
Example 2
The extraction method of berberine of the embodiment comprises the following steps:
1) drying Coptidis rhizoma at 80 deg.C for 2 hr, and pulverizing to 40 mesh to obtain Coptidis rhizoma powder;
2) taking the coptis powder prepared in the step 1) and an extraction solvent into a stainless steel reaction kettle with a polytetrafluoroethylene lining according to the proportion of adopting 60mL of extraction solvent for every 1g of coptis powder, and controlling the filling rate to be 60%; the adopted extraction solvent is a mixed solution of ethanol and water, and the volume concentration of the ethanol in the mixed solution is 70 percent;
3) sealing the reaction kettle, putting the reaction kettle in an oven with the temperature of 140 ℃, and extracting for 20 minutes;
4) and after extraction is finished, taking out the reaction kettle from the oven, naturally cooling to room temperature, opening the reaction kettle, and centrifugally separating the content in the polytetrafluoroethylene reaction kettle to obtain the berberine extracting solution.
Example 3
The extraction method of berberine of the embodiment comprises the following steps:
1) drying Coptidis rhizoma at 60 deg.C for 3 hr, and pulverizing to 60 mesh to obtain Coptidis rhizoma powder;
2) putting the coptis powder prepared in the step 1) and the extraction solvent into a stainless steel reaction kettle with a polytetrafluoroethylene lining according to the proportion of 40mL of extraction solvent adopted for every 1g of coptis powder, and controlling the filling rate to be 80%; the adopted extraction solvent is a mixed solution of ethanol and water, and the volume concentration of the ethanol in the mixed solution is 50 percent;
3) sealing the reaction kettle, putting the reaction kettle in an oven with the temperature of 120 ℃, and extracting for 50 minutes;
4) and after extraction is finished, taking out the reaction kettle from the oven, naturally cooling to room temperature, opening the reaction kettle, and centrifugally separating the content in the polytetrafluoroethylene reaction kettle to obtain the berberine extracting solution.
Example 4
The extraction method of berberine of the embodiment comprises the following steps:
1) drying Coptidis rhizoma at 60 deg.C for 3 hr, and pulverizing to 60 mesh to obtain Coptidis rhizoma powder;
2) taking the coptis powder prepared in the step 1) and an extraction solvent into a stainless steel reaction kettle with a polytetrafluoroethylene lining according to the proportion of 30mL of the extraction solvent adopted for every 1g of coptis powder, and controlling the filling rate to be 72%; the adopted extraction solvent is a mixed solution of ethanol and water, and the volume concentration of the ethanol in the mixed solution is 70 percent;
3) sealing the reaction kettle, putting the reaction kettle in an oven with the temperature of 120 ℃, and extracting for 50 minutes;
4) and after extraction is finished, taking out the reaction kettle from the oven, naturally cooling to room temperature, opening the reaction kettle, and centrifugally separating the content in the polytetrafluoroethylene reaction kettle to obtain the berberine extracting solution.
Example 5
The extraction method of berberine of the embodiment comprises the following steps:
1) drying Coptidis rhizoma at 70 deg.C for 3 hr, and pulverizing to 60 mesh to obtain Coptidis rhizoma powder;
2) taking the coptis powder prepared in the step 1) and an extraction solvent into a stainless steel reaction kettle with a polytetrafluoroethylene lining according to the proportion of 20mL of the extraction solvent adopted for every 1g of coptis powder, and controlling the filling rate to be 70%; the adopted extraction solvent is a mixed solution of ethanol and water, and the volume concentration of the ethanol in the mixed solution is 60 percent;
3) sealing the reaction kettle, placing the reaction kettle in an oven with the temperature of 110 ℃, and extracting for 60 minutes;
4) and after extraction is finished, taking out the reaction kettle from the oven, naturally cooling to room temperature, opening the reaction kettle, and centrifugally separating the content in the polytetrafluoroethylene reaction kettle to obtain the berberine extracting solution.
Example 6
The extraction method of berberine of the embodiment comprises the following steps:
1) drying Coptidis rhizoma at 70 deg.C for 3 hr, and pulverizing to 60 mesh to obtain Coptidis rhizoma powder;
2) taking the coptis powder prepared in the step 1) and an extraction solvent into a stainless steel reaction kettle with a polytetrafluoroethylene lining according to the proportion of adopting 30mL of the extraction solvent for every 1g of coptis powder, and controlling the filling rate to be 75%; the adopted extraction solvent is a mixed solution of ethanol and water, and the volume concentration of the ethanol in the mixed solution is 60 percent;
3) sealing the reaction kettle, putting the reaction kettle in an oven with the temperature of 130 ℃, and extracting for 40 minutes;
4) and after extraction is finished, taking out the reaction kettle from the oven, naturally cooling to room temperature, opening the reaction kettle, and centrifugally separating the content in the polytetrafluoroethylene reaction kettle to obtain the berberine extracting solution.
Example 7
The extraction method of berberine of the embodiment comprises the following steps:
1) drying raw material cortex Phellodendri at 70 deg.C for 3 hr, and pulverizing to 60 mesh to obtain cortex Phellodendri powder;
2) putting the phellodendron powder prepared in the step 1) and an extraction solvent into a stainless steel reaction kettle with a polytetrafluoroethylene lining according to the proportion of 40mL of the extraction solvent adopted for every 1g of phellodendron powder, and controlling the filling rate to be 80%; the adopted extraction solvent is a mixed solution of ethanol and water, and the volume concentration of the ethanol in the mixed solution is 40%;
3) sealing the reaction kettle, putting the reaction kettle in an oven with the temperature of 130 ℃, and extracting for 35 minutes;
4) and after extraction is finished, taking out the reaction kettle from the oven, naturally cooling to room temperature, opening the reaction kettle, and centrifugally separating the content in the polytetrafluoroethylene reaction kettle to obtain the berberine extracting solution.
Example 8
The extraction method of berberine of the embodiment comprises the following steps:
1) drying cortex Phellodendri and Coptidis rhizoma at 70 deg.C for 3 hr, and pulverizing to 60 mesh to obtain cortex Phellodendri and Coptidis rhizoma powder;
2) taking 40mL of extraction solvent according to the proportion of each 1g of mixed powder of golden cypress and golden thread (the mass ratio of golden thread to golden cypress is 1:1), putting the golden cypress and golden thread powder prepared in the step 1) and the extraction solvent into a stainless steel reaction kettle with a polytetrafluoroethylene lining, and controlling the filling rate to be 80%; the adopted extraction solvent is a mixed solution of ethanol and water, and the volume concentration of the ethanol in the mixed solution is 70 percent;
3) sealing the reaction kettle, putting the reaction kettle in an oven with the temperature of 120 ℃, and extracting for 50 minutes;
4) and after extraction is finished, taking out the reaction kettle from the oven, naturally cooling to room temperature, opening the reaction kettle, and centrifugally separating the content in the polytetrafluoroethylene reaction kettle to obtain the berberine extracting solution.
Comparative example 1
The extraction method of the berberine of the comparative example comprises the following steps:
1) drying Coptidis rhizoma at 60 deg.C for 3 hr, and pulverizing to 60 mesh to obtain Coptidis rhizoma powder;
2) placing the coptis powder prepared in the step 1) and an extraction solvent into a flask according to the proportion of 30mL of the extraction solvent to each 1g of the coptis powder; the adopted extraction solvent is a mixed solution of ethanol and water, and the concentration of the ethanol in the mixed solution is 70 percent;
3) mounting a reflux device, and carrying out reflux extraction for 50 minutes in a water bath at 80 ℃;
4) after extraction, naturally cooling to room temperature, and centrifugally separating the content in the flask to obtain the berberine extracting solution.
Examples of the experiments
1) Respectively taking the berberine extracting solutions prepared in the embodiments 1-8 and the comparative example 1 by adopting an ultraviolet spectrophotometry to detect the concentration of berberine in the berberine extracting solutions, calculating the mass of the berberine in the berberine extracting solutions prepared in the embodiments, and further calculating the extraction rate of the berberine, wherein the specific process is as follows:
i) drawing a standard curve: and (3) precisely weighing 5mg of berberine standard, dissolving with 50% methanol, and metering to a 10mL volumetric flask to obtain standard stock solution. Then taking the berberine stock solution to dilute the berberine stock solution into standard solutions of 2.5 mu g/mL, 5 mu g/mL, 7.5 mu g/mL, 10.0 mu g/mL and 12.5 mu g/mL, and detecting the solutions by an ultraviolet spectrophotometer. The absorbance at 345nm is plotted as ordinate (y) and the concentration of the standard substance is plotted as abscissa (x), and as shown in FIG. 1, the regression equation of the standard curve is that y is 0.0161x-0.0318, R2＝0.9993。
ii) calculation of berberine extraction: the berberine extraction rate (the mass of berberine in the total extract/the mass of the medicinal powder) is multiplied by 100 percent, and the result is shown in table 1.
2) Taking the berberine extracting solution of each example and comparative example to volatilize the solvent at normal temperature and drying the solvent to obtain berberine crude extract solid, and calculating the mass percentage content of the berberine in the berberine crude extract solid according to the measured berberine concentration in the berberine extracting solution, wherein the results are shown in table 1.
TABLE 1 extraction rates of berberine in examples 1 to 8 and
drying to obtain solid containing berberine
As can be seen from Table 1, the extraction rate of berberine is up to 8.53% at the maximum under the condition of extraction temperature of 120 ℃ for 50min, which is 1.4 times of that of ethanol reflux method under the same extraction conditions, when 30mL of extraction solvent is adopted correspondingly for every 1g of coptis powder, the ethanol concentration in the extraction solvent is 70%, and the extraction time is 50min, which is mainly because the extraction temperature is higher and the molecular motion is more vigorous under the condition of solvothermal, thereby increasing the exchange rate of the extraction solvent in and out of cells. Meanwhile, the closed extraction system improves the extraction pressure, so that the plant cell walls are broken or cracked more fully, and the dissolution resistance of the cell contents is reduced. Therefore, the solvothermal method has a higher extraction rate. In addition, the extraction method of the berberine can also improve the content of the berberine in the solute of the berberine extracting solution, and the berberine in the berberine crude extract obtained by volatilizing the solvent and drying the berberine extracting solution can reach 43 percent by mass.
Claims (6)
1. A method for extracting berberine is characterized by comprising the following steps: the method comprises the following steps: putting the traditional Chinese medicine powder and an extraction solvent into an extraction container, sealing the extraction container, extracting at 100-140 ℃, and performing solid-liquid separation after extraction to obtain a berberine extracting solution;
the extraction solvent is a mixed solution of ethanol and water, and the volume concentration of the ethanol in the extraction solvent is 30-70%; the traditional Chinese medicine powder is selected from one or two of coptis chinensis powder and phellodendron amurense powder.
2. The method for extracting berberine according to claim 1, wherein: the extraction time is 10-60 min at 100-140 ℃.
3. The method for extracting berberine according to claim 1, wherein: the volume of the extraction solvent adopted for each 1g of the traditional Chinese medicine powder is 10-60 mL.
4. The method for extracting berberine according to claim 1, wherein: the traditional Chinese medicine powder is obtained by drying traditional Chinese medicines and then crushing the dried traditional Chinese medicines; the drying treatment temperature is 50-80 ℃, and the drying treatment time is 2-6 h.
5. The method for extracting berberine according to claim 1, wherein: the particle size of the traditional Chinese medicine powder is 40-80 meshes.
6. The method for extracting berberine according to claim 1, wherein: the filling rate of the traditional Chinese medicine powder and the extraction solvent in the extraction container is 60-80%.
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| CN103149292A (en) * | 2013-02-22 | 2013-06-12 | 苏州大学 | Extraction method and detection method for berberine and palmatine in cortex phellodendri chinensis |
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| CN107892688A (en) * | 2017-11-15 | 2018-04-10 | 湘潭大学 | A kind of method that Berberine hydrochloride is prepared from golden cypress |
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| CN103149292A (en) * | 2013-02-22 | 2013-06-12 | 苏州大学 | Extraction method and detection method for berberine and palmatine in cortex phellodendri chinensis |
| CN104650071A (en) * | 2015-03-09 | 2015-05-27 | 丁素君 | Method for extracting berberine from rhizome of rhizoma coptidis |
| CN107892688A (en) * | 2017-11-15 | 2018-04-10 | 湘潭大学 | A kind of method that Berberine hydrochloride is prepared from golden cypress |
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