CN111072731A - Method for purifying and obtaining glycyrrhetate from liquorice - Google Patents
Method for purifying and obtaining glycyrrhetate from liquorice Download PDFInfo
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Abstract
The invention discloses a method for purifying and obtaining glycyrrhetate from liquorice. The method comprises mixing Glycyrrhrizae radix with organic solvent at a certain ratio, adding first pH regulator, controlling system pH, and performing multi-stage extraction of glycyrrhizic acid; and adding a second pH regulator into the primary extracting solution, controlling the pH value of the system, precipitating, filtering and washing the glycyrrhizic acid in a salt form to further purify the glycyrrhizic acid, wherein the filtrate generated in the purification process and the subsequent primary extracting solution can be continuously used for extracting the medicinal material of the liquorice in the next round/batch. The method realizes high-efficiency extraction, separation and purification of glycyrrhizic acid, and avoids the damage of glycyrrhizic acid; the used organic solvent can be efficiently recovered by modern means, the problems of wastewater treatment and environmental protection in the traditional glycyrrhizic acid extraction process are solved, the nutrient components such as starch, protein and the like in the licorice dregs are well preserved, and the efficient utilization of the licorice resource is realized.
Description
Technical Field
The invention relates to the technical field of medicinal material purification, in particular to a method for purifying and obtaining glycyrrhetate from liquorice.
Background
The licorice is the dried root and rhizome of glycyrrhiza (Glycyrrhiza uralensis Fisch.) of Glycyrrhiza of Leguminosae, Glycyrrhiza inflata (G.inflataBat.), Glycyrrhiza glabra (G.Glabral.), is a commonly used bulk drug and is reputed as "nineteen herbs" and "old people". The Glycyrrhrizae radix contains various components such as saponins, triterpenes, flavonoids, coumarin, sterol, alkaloid, volatile oil, organic acid, amino acids, etc. Glycyrrhizic acid is a sweet component of liquorice, also called glycyrrhizin, and has the sweetness 200-300 times that of cane sugar. Glycyrrhizic acid has the efficacies of antianaphylaxis, anti-tumor, antiulcer, anti-inflammation and the like, and is the pentacyclic triterpenoid saponin compound which is most and earliest researched in the liquorice components at present. Glycyrrhizic acid is widely applied to the fields of medicines, foods and cosmetics mainly in the forms of ammonium salt, potassium salt, zinc salt, glycyrrhetinic acid and the like.
The glycyrrhizic acid content in the licorice root rhizome is about 1-12%, and the glycyrrhizic acid mainly exists in the forms of free acid, sodium salt, calcium salt, potassium salt and magnesium salt. The extraction method of glycyrrhizic acid mainly comprises water extraction, dilute ammonia water extraction, ammonia alcohol extraction, microwave-assisted extraction, etc. Because of low solvent cost, safe application and high extraction rate of effective components, the dilute ammonia water extraction method is the main glycyrrhizic acid extraction method in the current industrial production. However, ammonia water is used in the extraction process, so that on one hand, the extracted decoction dregs contain a large amount of ammonia, the decoction dregs are smelly in the storage process, the nutrient components such as starch in the medicinal materials are damaged, and the subsequent application of the decoction dregs is greatly limited; on the other hand, the process adopts an alkali dissolution and acid precipitation process to obtain glycyrrhizic acid powder, the strong acid solution has about 20 percent of destructive effect on glycyrrhizic acid, and a large amount of acid water generated by the process brings great pressure to environmental protection. At present, most domestic glycyrrhizic acid powder manufacturers stop due to the environmental protection requirements and the limitation of cost improvement.
Along with the year-by-year shortage of liquorice resources in China in recent years and the strong advocated national aspects of green manufacturing and environmental protection, the extraction of the liquorice needs to be developed in a clean and efficient extraction process.
The information disclosed in this background section is only for enhancement of understanding of the general background of the invention and should not be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person skilled in the art.
Disclosure of Invention
The invention aims to provide a method for purifying and obtaining glycyrrhetate from liquorice, which realizes high-efficiency extraction, separation and purification of glycyrrhizic acid and avoids the damage of glycyrrhizic acid; the used organic solvent can be efficiently recovered by modern means, the problems of wastewater treatment and environmental protection in the traditional glycyrrhizic acid extraction process are solved, the nutrient components such as starch, protein and the like in the licorice dregs are well preserved, and the efficient utilization of the licorice resource is realized.
In order to achieve the purpose, the invention provides a method for purifying and obtaining glycyrrhetate from liquorice, which comprises the following steps: crushing liquorice: crushing a licorice medicinal material into licorice medicinal powder, and controlling the granularity of the licorice medicinal powder to be 30-200 meshes; primary extraction: mixing organic solvent and water to obtain first solvent, and mixing with the above Glycyrrhrizae radix powder in a ratio of (2-20): 1 (volume/weight, V/W), stirring, adding a first pH regulator, regulating the pH value, stirring and extracting for 0.5-3 hours, filtering, and collecting a first-level extracting solution and dregs; and purification: adding a second pH regulator into the primary extracting solution, controlling the pH value of the system, converting glycyrrhizic acid in the primary extracting solution into glycyrrhetate, precipitating from an extracting solution system of an organic solvent, filtering, collecting precipitate, washing the precipitate with a washing solution, filtering, respectively collecting the filtered washing solution and the precipitate, drying the washed precipitate at 50-80 ℃ to obtain the glycyrrhetate, wherein the total yield of the glycyrrhetate obtained by the purification method is not lower than 90%.
In an embodiment of the present invention, in the licorice crushing step, the grain size of the licorice powder is preferably 30 to 100 mesh.
The invention also provides a method for purifying and obtaining glycyrrhetate from liquorice, which comprises the following steps: crushing liquorice: crushing a licorice medicinal material into licorice medicinal powder, and controlling the granularity of the licorice medicinal powder to be 30-200 meshes; primary extraction: mixing organic solvent and water to obtain first solvent, and mixing with the Glycyrrhrizae radix powder in a ratio of (2-20): 1(V/W), stirring, adding a first pH regulator, regulating the pH value, stirring and extracting for 0.5-3 hours, filtering, and collecting a first-stage extracting solution and dregs; and (3) K-level extraction: mixing the first solvent and the residue obtained in the previous stage of extraction with a solvent of (2-20): 1(V/W), stirring, adding a first pH regulator, regulating the pH value, stirring and extracting for 0.5-3 hours, filtering, and collecting a K-grade extracting solution and K-grade medicine residues; performing the K-stage extraction starting from K-2 up to N-stage extraction, thereby completing a complete round of extraction, N being 2 or more and 8 or less, and purifying: adding a second pH regulator into the primary extracting solution, controlling the pH value of the system, filtering, collecting precipitate, washing the precipitate with a washing solution, filtering, respectively collecting the filtered washing solution and the precipitate, and drying the washed precipitate at 50-80 ℃ to obtain the glycyrrhetate.
In an embodiment of the present invention, in each round of extraction process, the first solvent further includes a mixed solution composed of each stage of extraction solution in the previous round/batch of extraction except the first stage of extraction solution and the filtered washing solution in the purification step, and this measure can recycle the solution, thereby avoiding the problems of wastewater treatment and environmental protection in the conventional glycyrrhizic acid extraction process.
In an embodiment of the present invention, in the primary extraction step, the organic solvent is at least one selected from alcohols, alcohol derivatives, and ketones, and preferably, the organic solvent is at least one selected from methanol, ethanol, ethyl acetate, and acetone.
In an embodiment of the present invention, in the primary extraction step, the water accounts for 0 to 30% by volume of the first solvent.
In one embodiment of the present invention, in the primary extraction step, the first solvent is selected from one of methanol-ethanol-water and ethanol-ethyl acetate-water.
In an embodiment of the present invention, in the primary extraction step, the ratio of the first solvent to the licorice powder is preferably (4-9): 1 (volume/weight, V/W).
In one embodiment of the present invention, in the first extraction step and the K-extraction step, the first pH adjusting agent is at least one selected from the group consisting of: h2SO4、HCl、H3PO4Polyphosphoric acid, CH3COOH, citric acid, oxalic acid, cation exchange resin, a buffer system with pH less than or equal to 2, and solutions with different concentrations prepared by at least one of the substances; preferably, the first pH adjusting agent is sulfuric acid-hydrochloric acid. The preferable buffering system with the pH value less than or equal to 2 is glycine-hydrochloric acid buffer solution; further preferably 0.1 to 0.3M glycine-hydrochloric acid buffer solution.
In one embodiment of the present invention, in the first extraction step and the K-extraction step, the pH is adjusted to 4 or less, preferably to 0.5-4.
In one embodiment of the present invention, in the first extraction step and the K-stage extraction step, the time for stirring extraction after adding the first pH adjuster is preferably 0.3 to 1 hour.
In one embodiment of the present invention, in the purification step, the second pH adjusting agent is at least one selected from the group consisting of: metal, metal oxide, metal hydroxide, salt formed by metal, anion exchange resin, ammonia gas, liquid ammonia, ammonia water, ammonium salt, triethylamine, a buffer system with a pH buffer range of 9-13, and a solution prepared from at least one of the substances with various concentrations; wherein the metal is metal sodium, potassium, calcium, magnesium and zinc; the metal oxide is the oxide of sodium, potassium, calcium, magnesium and zinc; the metal salt is various acid salts formed by sodium, potassium, calcium, magnesium and zinc; the metal hydroxide is hydroxide of sodium, potassium, calcium, magnesium and zinc. In the actual operation process, the corresponding substances or solutions are selected according to the type of the glycyrrhetate finally required.
In one embodiment of the present invention, in the purification step, the pH is controlled to be between 5 and 10.
In one embodiment of the present invention, in the purification step, the washing solution is the same as the first solvent, and preferably, the washing solution is one selected from methanol-ethanol-water and ethanol-ethyl acetate-water.
In one embodiment of the present invention, in the purification step, the washing solution is washed at normal temperature or under heating; preferably, the heating temperature is 40-80 ℃, and the washing times of the washing solution are 2-3 times.
In an embodiment of the present invention, in the purifying step, the washed precipitate is preferably dried at 50 to 70 ℃ for standby.
Compared with the prior art, the invention has the following beneficial effects:
the method for purifying and obtaining glycyrrhetate from liquorice uses an organic solvent and a first pH regulator to extract glycyrrhizic acid, simultaneously uses a salt precipitation mode to precipitate glycyrrhizic acid, and further purifies the glycyrrhizic acid, and filtrate generated in the purification process and subsequent glycyrrhizic acid extracting solution can be continuously applied to the extraction of the liquorice medicinal material in the next round/batch. On one hand, the extraction rate of the glycyrrhizic acid is improved, the loss and damage of the alkali dissolution and acid precipitation step in the traditional process are avoided, and the damage of the glycyrrhizic acid is avoided; on the other hand, the extraction solvent used in the extraction process is recycled to extract medicinal materials, so that the utilization rate of the organic solvent is improved, and meanwhile, the organic solvent is efficiently recovered by modern means, so that the problems of wastewater treatment and environmental protection in the traditional glycyrrhizic acid extraction process are solved, the nutritional ingredients such as starch, protein and the like in the licorice dregs are well preserved, and the whole-herb efficient utilization of the licorice resource is realized.
Drawings
FIG. 1 is a process flow diagram of a method for purifying glycyrrhetate from licorice root according to an embodiment of the present invention.
Detailed Description
The following detailed description of specific embodiments of the invention is provided, but it should be understood that the scope of the invention is not limited to the specific embodiments.
Throughout the specification and claims, unless explicitly stated otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated element or component but not the exclusion of any other element or component.
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The method for purifying glycyrrhetate from licorice is described in the following examples.
Example 1
(1) Crushing and crushing a licorice medicinal material to ensure that the granularity of the medicinal powder is all 40-mesh standard sieve;
(2) primary extraction: taking 1kg of the licorice powder, adding 5L of methanol-ethanol-water (V/V/V, 10/85/5), stirring uniformly, adding a proper amount of 5M sulfuric acid-hydrochloric acid (V/V, 1/1), controlling the pH value of a mixed system to be 2-4, stirring and extracting for 30 minutes, filtering, and respectively collecting a level 1 extracting solution and dregs;
(3) two-six stage extraction: extracting the medicinal materials by the same method for five stages, namely 6 stages, and collecting the extraction solution and the dregs respectively;
(4) and (3) separating and purifying glycyrrhizic acid: adding 0.1M ammonia-ammonium chloride mixed solution (V/V, 4/1) into the primary extractive solution, controlling pH to 5-10 to generate glycyrrhetate precipitate, filtering, collecting precipitate, washing the precipitate with methanol-ethanol-water (V/V/V, 10/85/5) at room temperature for 3 times, filtering, and collecting precipitate and filtrate respectively; drying the precipitate at 60 ℃ to obtain ammonium glycyrrhetate;
(5) recycling of the solution: the 2-6 grade extracting solution and the glycyrrhetate washing filtrate are used for extracting a new round of medicinal materials.
Example 2
(1) Crushing and crushing a licorice medicinal material to ensure that the granularity of the medicinal powder is all 40-mesh standard sieve;
(2) primary extraction: taking 1kg of the licorice powder, adding 5L of the grade 2-6 extract and the washed glycyrrhetate filtrate obtained in the example 1, uniformly stirring, adding a proper amount of 5M sulfuric acid-hydrochloric acid (V/V, 1/1), controlling the pH value of a mixed system to be 2-4, stirring and extracting for 30 minutes, filtering, and respectively collecting grade 1 extract and dregs;
(3) two-six stage extraction: extracting the medicinal materials by the same method for five stages, namely 6 stages, and collecting the extraction solution and the dregs respectively;
(4) and (3) separating and purifying glycyrrhizic acid: adding 0.1M ammonia-ammonium chloride mixed solution (V/V, 4/1) into the primary extractive solution, controlling pH to 5-10 to generate glycyrrhetate precipitate, filtering, collecting precipitate, washing the precipitate with methanol-ethanol-water (V/V/V, 10/85/5) at room temperature for 3 times, filtering, and collecting precipitate and filtrate respectively; drying the precipitate at 60 ℃ to obtain ammonium glycyrrhetate;
(5) recycling of the solution: the 2-6 grade extracting solution and the glycyrrhetate washing filtrate are used for extracting a new round of medicinal materials.
Example 3
(1) Crushing and crushing a licorice medicinal material to ensure that the granularity of the medicinal powder is all through a standard sieve of 80 meshes;
(2) primary extraction: taking 1kg of the licorice powder, adding 8L of ethanol-water (V/V, 9010), stirring uniformly, adding a proper amount of 0.2M glycine-hydrochloric acid (V/V, 1/4) solution, controlling the pH value of the mixed system to be between 2 and 4, stirring and extracting for 30 minutes, filtering, and respectively collecting a level 1 extracting solution and dregs;
(3) second-fifth stage extraction: extracting the medicinal materials by four stages, namely 5 stages, by the same method, and respectively collecting the extracting solution and the dregs of a decoction at each stage;
(4) and (3) separating and purifying glycyrrhizic acid: adding 2M potassium hydroxide solution into the first-stage extractive solution, controlling pH to 5-10 to obtain potassium glycyrrhetate precipitate, filtering, collecting precipitate, washing the precipitate with ethanol-water (V/V, 90/10) at room temperature for 2 times, filtering, and collecting precipitate and filtrate respectively; precipitating and drying at 60 ℃ to obtain potassium glycyrrhetate;
(5) recycling of the solution: the 2-5 grade extracting solution and the glycyrrhetate washing filtrate are used for extracting a new round of medicinal materials.
Example 4
(1) Crushing and crushing a licorice medicinal material to ensure that the granularity of the medicinal powder is all through a standard sieve of 80 meshes;
(2) primary extraction: taking 1kg of the licorice powder, adding 8L of the grade 2-6 extract and the washed glycyrrhetate filtrate obtained in the example 1, uniformly stirring, adding a proper amount of 0.2M glycine-hydrochloric acid (V/V, 1/4), controlling the pH value of a mixed system to be 2-4, stirring and extracting for 30 minutes, filtering, and respectively collecting the grade 1 extract and dregs of a decoction;
(3) second-fifth stage extraction: extracting the medicinal materials by four stages, namely 5 stages, by the same method, and respectively collecting the extracting solution and the dregs of a decoction at each stage;
(4) and (3) separating and purifying glycyrrhizic acid: adding 2M potassium hydroxide solution into the first-stage extractive solution, controlling pH to 5-10 to obtain potassium glycyrrhetate precipitate, filtering, collecting precipitate, washing the precipitate with ethanol-water (V/V, 90/10) at room temperature for 2 times, filtering, and collecting precipitate and filtrate respectively; precipitating and drying at 60 ℃ to obtain potassium glycyrrhetate;
(5) recycling of the solution: the 2-5 grade extracting solution and the glycyrrhetate washing filtrate are used for extracting a new round of medicinal materials.
The present invention further provides statistics on the extraction rate of glycyrrhizic acid in examples 1-4 above.
Example 1 and example 2 Glycyrrhetinic acid extraction statistics
The detection method comprises the following steps: high performance liquid chromatography
(1) Chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile-0.01 mol/L phosphoric acid solution (38: 62) is used as a mobile phase; the detection wavelength is 252 nm; the flow rate is 1 ml/min; sample introduction amount: 10 mu L of the solution; the theoretical plate number is not lower than 2000 calculated according to the ammonium glycyrrhizinate peak, and the separation degree of the ammonium glycyrrhizinate peak and the internal standard substance peak is in accordance with the requirement.
(2) Preparation of internal standard solution: taking about 70mg of n-butyl p-hydroxybenzoate, accurately weighing, placing in a 100ml measuring flask, dissolving with diluted ethanol, diluting to scale, and shaking.
(3) Preparation of control solutions: precisely weighing about 20mg of monoammonium glycyrrhizinate as reference substance, placing into a 100ml measuring flask, adding diluted ethanol for dissolving, precisely adding 5ml of internal standard solution, diluting with diluted ethanol to scale, and shaking up.
(4) Sample solution preparation
Primary extracting solution: precisely transferring 2mL to 100mL volumetric flasks, precisely adding 5mL of internal standard solution, and diluting with diluted ethanol to a scale;
second-sixth stage extract: precisely transferring 5mL to 100mL volumetric flasks respectively, precisely adding 5mL of internal standard solution, and diluting with diluted ethanol to a scale;
ammonium glycyrrhetate salt: precisely weighing 25mg of glycyrrhizic acid intermediate, transferring to a 100mL volumetric flask, precisely adding 5mL of internal standard solution, and diluting to scale with diluted ethanol.
The glycyrrhizic acid concentration and extraction rate of each stage of the extract liquid in example 1 and example 2 were counted, and the data are shown in table 1.
Table 1 glycyrrhizic acid extraction statistics of example 1 and example 2
As shown in Table 1, after the licorice root medicinal material is extracted by the method for 6 grades, the glycyrrhizic acid in the licorice root medicinal material is basically and completely extracted. In the process, the primary extracting solution is used for precipitating the ammonium glycyrrhetate, and the transfer rate of the glycyrrhetate relative to the primary extracting solution is more than 94 percent and is higher than that of the glycyrrhizic acid by the traditional extracting process. According to the invention, the solvent is recycled to extract the medicinal materials, and the concentration of the primary extracting solution is gradually increased along with the increase of the application times, so that the yield of glycyrrhetate obtained by each round of extraction of the medicinal materials is improved relative to the yield of the medicinal materials, and finally, the total yield of ammonium glycyrrhetate is over 90 percent during each round of extraction.
(II) Glycyrrhetinic acid extraction statistics of examples 3 and 4
The detection method comprises the following steps: high performance liquid chromatography
(1) Chromatographic conditions and system applicability test: octadecylsilane chemically bonded silica is used as a filling agent; acetonitrile-0.01 mol/L phosphoric acid solution (38: 62) is used as a mobile phase; the detection wavelength is 252 nm; the flow rate is 1 ml/min; sample introduction amount: 10 mu L of the solution; the theoretical plate number is not lower than 2000 calculated according to the ammonium glycyrrhizinate peak, and the separation degree of the ammonium glycyrrhizinate peak and the internal standard substance peak is in accordance with the requirement.
(2) Preparation of internal standard solution: taking about 70mg of n-butyl p-hydroxybenzoate, accurately weighing, placing in a 100ml measuring flask, dissolving with diluted ethanol, diluting to scale, and shaking.
(3) Preparation of control solutions: precisely weighing about 20mg of monoammonium glycyrrhizinate as reference substance, placing into a 100ml measuring flask, adding diluted ethanol for dissolving, precisely adding 5ml of internal standard solution, diluting with diluted ethanol to scale, and shaking up.
(4) Sample solution preparation
Primary extracting solution: precisely transferring 2mL to 100mL volumetric flasks, precisely adding 5mL of internal standard solution, and diluting with diluted ethanol to a scale;
second-fifth grade extract: precisely transferring 5mL to 100mL volumetric flasks respectively, precisely adding 5mL of internal standard solution, and diluting with diluted ethanol to a scale;
potassium glycyrrhetate: precisely weighing 25mg of glycyrrhizic acid intermediate, transferring to a 100mL volumetric flask, precisely adding 5mL of internal standard solution, and diluting to scale with diluted ethanol.
The glycyrrhizic acid concentration and extraction rate of each stage of the extract liquid in example 3 and example 4 were counted, and the data are shown in table 2.
Table 2 example 3 and example 4 glycyrrhizic acid extraction statistics
As shown in the table above, after the licorice medicinal material is extracted by the method for 6 grades, the glycyrrhizic acid is basically and completely extracted. In the process, the primary extracting solution is used for precipitating the ammonium glycyrrhetate, and the yield of the potassium glycyrrhetate is more than 94 percent relative to the transfer rate of the primary extracting solution, which is higher than that of the traditional extracting process of the glycyrrhizic acid. According to the invention, the solvent is recycled to extract the medicinal materials, and the concentration of the primary extracting solution is gradually increased along with the increase of the application times, so that the yield of potassium glycyrrhetate obtained by each round of extraction of the medicinal materials is improved relative to the medicinal materials, and finally, the total yield of potassium glycyrrhetate is more than 90% in each round of extraction.
The invention further carries out specific verification tests on the test conditions, and details are shown in the following.
(1) Granularity of medicinal materials:
pulverizing Glycyrrhrizae radix (glycyrrhizic acid content of 3.14%) into powder of 10 mesh, 30 mesh, 60 mesh, 80 mesh, 100 mesh and 200 mesh, respectively 50 g;
adding 250mL of methanol-ethanol-water (V/V/V, 10/85/5) and a proper amount of 5M sulfuric acid-hydrochloric acid (V/V, 1/1) into each medicinal material, stirring, extracting for 30min, filtering, collecting filtrate, recording the volume of the extract, detecting the concentration of the primary extract by a method, and counting the extraction condition of glycyrrhizic acid of the licorice medicinal material by particle size.
TABLE 3 particle size selection test results for Licorice root powder
| Sample (I) | No. 1-10 mesh | No. 2-30 mesh | No. 3-60 mesh | No. 4-80 mesh | No. 5-100 mesh | No. 6-200 mesh |
| Extraction rate | 42.94％ | 60.28％ | 65.77％ | 72.98％ | 74.96％ | 75.19％ |
As can be seen from Table 3, the mesh number of the licorice powder was controlled within a reasonable range to obtain a better extraction rate of glycyrrhizic acid.
(2) Extraction time
Crushing a licorice medicinal material to 80 meshes, taking 5 parts, 100 g/part, adding 500mL of methanol-ethanol-water (V/V/V, 10/85/5) and a proper amount of 5M sulfuric acid-hydrochloric acid (V/V, 1/1) respectively, stirring, extracting, performing primary extraction for 15min, 30min, 60min, 90min, 120min and 180min respectively, filtering, collecting filtrate, recording the volume of the extract, detecting the concentration of the primary extract according to a method, and counting the extraction conditions of the licorice medicinal material glycyrrhizic acid at different extraction time.
TABLE 4 first-order extraction selection test results
As can be seen from table 4, the primary extraction time of the glycyrrhiza uralensis powder is controlled within a reasonable range to obtain a better extraction rate of glycyrrhizic acid.
(3) Extraction order
Taking a licorice medicinal material, crushing to 100 meshes, 100kg, adding 500L ethanol-water (V/V; 90/10) in primary extraction, adding a proper amount of 5M sulfuric acid-hydrochloric acid (V/V, 1/1), stirring, extracting for 30min, filtering, collecting the volume of primary extract, and extracting at 2-5 stages: and (3) extracting the medicinal materials by four stages by adopting the same method, namely extracting 5 stages together, respectively collecting extracting solutions of all stages, recording the volumes of the extracting solutions, detecting the concentrations of the extracting solutions of all stages according to a method, and counting the extraction conditions of the glycyrrhizic acid of the licorice medicinal material by the extraction stages.
TABLE 5 results of extraction order selection test
| Order of the order | Concentration (%) | Extraction ratio (%) | Integrated extraction (%) |
| 1 | 7.62 | 84.25％ | 84.25％ |
| 2 | 1.15 | 12.14％ | 96.39％ |
| 3 | 0.22 | 2.39％ | 98.78％ |
| 4 | 0.06 | 0.64％ | 99.42％ |
| 5 | 0.02 | 0.18％ | 99.60％ |
As can be seen from table 5, the extraction grade of the glycyrrhiza uralensis powder is controlled within a reasonable range to obtain a better extraction rate of glycyrrhizic acid.
(4) Acidity of the extract
The method comprises the steps of crushing a licorice medicinal material to 80 meshes, taking 5 parts of the licorice medicinal material, taking 100 g/part of the licorice medicinal material, adding 500mL of methanol-ethanol-water (V/V/V, 10/85/5) and a proper amount of 5M sulfuric acid-hydrochloric acid (V/V, 1/1) respectively, controlling the pH value of a system to be about 1.0, 1.5, 2.0, 2.5, 3, 4, 5 and 6 respectively, stirring, extracting for 30min, filtering, collecting filtrate, recording the volume of the extract, detecting the concentration of the extract according to a method, and counting the influence of different acidity on extraction of glycyrrhizic acid of the licorice medicinal material.
TABLE 6 acidity selection test results for the extract
| Numbering | pH of the system | Concentration (mg/mL) | Extraction ratio (%) |
| 1 | 1.10 | 5.83％ | 76.7％ |
| 2 | 1.53 | 5.92％ | 77.9％ |
| 3 | 2.08 | 5.76％ | 76.2％ |
| 4 | 2.61 | 5.87％ | 77.2％ |
| 5 | 3.04 | 5.62％ | 73.9％ |
| 6 | 4.21 | 2.34％ | 30.8％ |
| 7 | 5.02 | 1.01％ | 13.3％ |
| 8 | 6.11 | 0.11％ | 1.5％ |
As can be seen from Table 6, the pH of the system was controlled within a reasonable range to ensure a suitable extraction yield of the drug.
(5) Determination of pH value in preparation of glycyrrhetate
Taking 5 parts of the primary extract of the licorice root, adding a proper amount of 0.1M ammonia-ammonium chloride mixed solution (V/V, 4/1) into each 200ml of the primary extract, respectively adjusting the pH value of the system to about 5.0, 6.0, 7.0, 8.0, 9.0 and 10.0, respectively, filtering, collecting filtrate, and detecting the ratio of the glycyrrhetate residue in the filtrate.
Table 7 preparation of glycyrrhetate pH selection test results
As can be seen from Table 7, the pH of the system was controlled within a reasonable range to precipitate more glycyrrhetate and to keep the filtrate residual concentration as low as possible.
The foregoing descriptions of specific exemplary embodiments of the present invention have been presented for purposes of illustration and description. It is not intended to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to explain certain principles of the invention and its practical application to enable one skilled in the art to make and use various exemplary embodiments of the invention and various alternatives and modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the claims and their equivalents.
Claims (10)
1. A method for purifying and obtaining glycyrrhetate from liquorice is characterized by comprising the following steps:
crushing liquorice: crushing a licorice medicinal material into licorice medicinal powder, and controlling the granularity of the licorice medicinal powder to be 30-200 meshes;
primary extraction: mixing organic solvent and water to obtain first solvent, and mixing with the Glycyrrhrizae radix powder in a ratio of (2-20): 1(V/W), stirring, adding a first pH regulator, regulating the pH value, stirring and extracting for 0.5-3 hours, filtering, and collecting a first-stage extracting solution and dregs; and
and (3) purification: adding a second pH regulator into the primary extracting solution, controlling the pH value of the system, filtering, collecting precipitate, washing the precipitate with a washing solution, filtering, respectively collecting the filtered washing solution and the precipitate, and drying the washed precipitate at 50-80 ℃ to obtain the glycyrrhetate.
2. A method for purifying and obtaining glycyrrhetate from liquorice is characterized by comprising the following steps:
crushing liquorice: crushing a licorice medicinal material into licorice medicinal powder, and controlling the granularity of the licorice medicinal powder to be 30-200 meshes;
primary extraction: mixing organic solvent and water to obtain first solvent, and mixing with the Glycyrrhrizae radix powder in a ratio of (2-20): 1(V/W), stirring, adding a first pH regulator to regulate the pH value, stirring and extracting for 0.5-3 hours, filtering, and collecting a first-stage extracting solution and dregs;
and (3) K-level extraction: mixing the first solvent and the residue obtained in the previous stage of extraction with a solvent of (2-20): 1(V/W), stirring, adding a first pH regulator, regulating the pH value, stirring and extracting for 0.5-3 hours, filtering, and collecting a K-grade extracting solution and K-grade medicine residues;
performing the K-level extraction starting from K-2 until N levels of extraction, thereby completing a complete round of extraction, N being greater than or equal to 2 and less than or equal to 8, and
and (3) purification: adding a second pH regulator into the primary extracting solution, controlling the pH value of the system, filtering, collecting precipitate, washing the precipitate with a washing solution, filtering, respectively collecting the filtered washing solution and the precipitate, and drying the washed precipitate at 50-80 ℃ to obtain the glycyrrhetate.
3. The method of claim 2, wherein the first solvent further comprises a mixture of the extract from each stage of the previous extraction, except the first stage of the extraction, and the filtered washing solution of the purification step during each extraction cycle.
4. The method for purifying glycyrrhetate according to claim 2, wherein the organic solvent is at least one selected from the group consisting of alcohols, alcohol derivatives and ketones, preferably at least one selected from the group consisting of methanol, ethanol, ethyl acetate and acetone.
5. The method of claim 2, wherein the water is present in the first solvent in an amount of 0-30% by volume of the first solvent in the first extraction step.
6. The method of claim 2, wherein in the first extraction step and the K extraction step, the first pH regulator is at least one selected from the group consisting of: h2SO4、HCl、H3PO4Polyphosphoric acid, CH3COOH, citric acid, oxalic acid, cation exchange resin, a buffer system with pH less than or equal to 2, and solutions with different concentrations prepared from at least one of the substances.
7. The method for purifying glycyrrhetate from licorice root as claimed in claim 2, wherein the pH value is adjusted to 4 or less in the first extraction and K extraction steps.
8. The method of claim 2, wherein the second pH adjusting agent is at least one selected from the group consisting of: metal, metal oxide, metal hydroxide, salt formed by metal, anion exchange resin, ammonia gas, liquid ammonia, ammonia water, ammonium salt, triethylamine, a buffer system with a pH buffer range of 9-13, and a solution prepared from at least one of the substances with various concentrations; wherein the metal is metal sodium, potassium, calcium, magnesium and zinc; the metal oxide is the oxide of sodium, potassium, calcium, magnesium and zinc; the salt formed by the metal is various acid salts formed by sodium, potassium, calcium, magnesium and zinc; the metal hydroxide is hydroxide of sodium, potassium, calcium, magnesium and zinc.
9. The method for purifying glycyrrhetate according to claim 2, wherein the pH value is controlled between 5 and 10 during the purification step.
10. The method for purifying glycyrrhetate according to claim 2, wherein the washing solution is the same as the first solvent and is washed at room temperature or under heating; preferably, the heating temperature is 40-80 ℃, and the washing times are 2-3.
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| WO2021134407A1 (en) * | 2019-12-31 | 2021-07-08 | 中国医药健康产业股份有限公司 | Method for extracting crude product of glycyrrhizic acid and glycyrrhizic flavone from licorice and purifying to obtain glycyrrhizinate |
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| WO2021134407A1 (en) * | 2019-12-31 | 2021-07-08 | 中国医药健康产业股份有限公司 | Method for extracting crude product of glycyrrhizic acid and glycyrrhizic flavone from licorice and purifying to obtain glycyrrhizinate |
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