CN111393499A - Method for removing triterpenoid impurities in diammonium glycyrrhizinate bulk drug - Google Patents
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Abstract
The invention belongs to the technical field of preparation of glycyrrhetate, and particularly relates to a method for removing triterpenoid impurities in a diammonium glycyrrhizinate raw material drug2And glycyrrhizin H2The diammonium glycyrrhizinate product with higher purity is obtained, and the obtained product has good color property, low content of related substances and high content of main components. Compared with the prior art, the method can purposefully remove impurities by utilizing the difference between the properties of related substances and main components, adopts liquid chromatography to detect the content of the related substances in the diammonium glycyrrhizinate bulk drug and the preparation in real time, and is beneficial to improving the diammonium glycyrrhizinate sourceQuality standards of the materials and the preparations.
Description
Technical Field
The invention belongs to the technical field of preparation of glycyrrhetate, and particularly relates to a method for removing triterpenoid impurities in diammonium glycyrrhizinate bulk drugs.
Background
The diammonium glycyrrhizinate is 18 α -diammonium glycyrrhizinate, is an effective component extracted from traditional Chinese medicine licorice, is a renewal product of monoammonium glycyrrhizinate, is a third-generation extract of the effective component of traditional Chinese medicine licorice, is a chronic hepatitis treatment drug with strong pharmacological activity, has strong effects of resisting inflammation, protecting liver cell membranes and improving liver functions, has prevention and treatment effects on liver injury caused by various hepatotoxic agents, presents a certain dose dependence, and can obviously improve the survival rate and improve the liver functions on chronic liver injury caused by compound pathogenic factors.
Related substances in the diammonium glycyrrhizinate bulk drug mainly refer to starting materials, intermediates and byproducts brought in the production process, degradation products generated in the storage process and the like, and comprise triterpenoid impurities. The preparation process of the diammonium glycyrrhizinate comprises the following steps: ammonium glycyrrhetate or triammonium glycyrrhetate is refluxed by alkali liquor to complete configuration conversion, then undergoes pH adjustment and organic solvent extraction layering, ammonia gas or ammonia water is added to synthesize a diammonium glycyrrhetate crude product, and then the ammonium glycyrrhetate crude product is refined for multiple times to reduce various impurities.
The synthetic route of diammonium glycyrrhizinate is shown as the following formula:
the related substances (triterpenoids) in the diammonium glycyrrhizinate bulk drug refer to impurity peaks in front of and behind a main peak of diammonium glycyrrhizinate on a liquid chromatogram, wherein the impurity peak in front of the main peak is glycyrrhizin G2Is prepared from ammonium glycyrrhizinate or glycyrrhizic acidThe impurities can be effectively reduced by adjusting the concentration of the alkali liquor in the reflux process of the alkali liquor when the crude product is synthesized by introducing the triammonium salt; the impurity peak behind the main peak is glycyrrhizin H2The impurities are generated in the reflux process, and can be reduced by refining for multiple times in the later period.
At present, the research on related substances in diammonium glycyrrhizinate bulk drugs and preparations sold in the market only stays at the stage of detecting single maximum impurity, secondary large impurity and total amount of impurities, and no specific structural identification report research is found. The method adopts the liquid chromatography-mass spectrometry technology to firstly determine the structure of the impurity, and then adopts a recrystallization method to remove the impurity according to the structure and the property of the impurity, thereby effectively reducing the triterpenoid impurity glycyrrhizin G in the diammonium glycyrrhizinate2And glycyrrhizin H2The content of (a).
Disclosure of Invention
In order to solve the technical problems, the invention provides a method for removing triterpenoid impurities in diammonium glycyrrhizinate bulk drugs.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for removing triterpenes impurities in diammonium glycyrrhizinate raw material medicine, wherein the triterpenes impurities comprise glycyrrhizin G2And glycyrrhizin H2The removing method comprises the following steps:
a: glycyrrhizasaponin G2Removing, namely adding acetic acid solution I into a diammonium glycyrrhizinate crude product, heating to 85-95 ℃, reacting for 25-35min, cooling to room temperature, standing to separate out crystals for 66-72h, centrifuging, taking a filter cake, washing the filter cake with the acetic acid solution I, centrifuging, draining, adding ethanol solution I with 2-3 times of the mass of the filter cake, pulping at room temperature, centrifuging, washing the filter cake with the ethanol solution I, centrifuging, draining the filter cake, and drying in vacuum to obtain a diammonium glycyrrhizinate primary refined product; adding acetic acid solution II which is 3-5 times of the weight of the primary refined diammonium glycyrrhizinate into the primary refined diammonium glycyrrhizinate, heating to 96-97 ℃, reacting for 20-30min, adding active carbon, and preservingDecolorizing at room temperature for 20-30min, removing carbon while hot, filtering, standing the filtrate at 0-10 deg.C for crystallization for 66-72 hr, centrifuging, filtering, rinsing with acetic acid solution II, filtering to obtain filter cake, and vacuum drying to obtain refined diammonium glycyrrhizinate;
b: glycyrrhizin H2And (3) removing, namely heating the ethanol solution II to 85-90 ℃, adding 15-18g of the diammonium glycyrrhizinate secondary refined product obtained in the step (b), continuously heating to 75-81 ℃, adding 0.6-1.0g of active carbon under the condition of 78 +/-3 ℃, carrying out reflux decoloring for 10-20min at 78 +/-3 ℃, filtering, cooling the filtrate, refrigerating and crystallizing for 3-4d, filtering the crystallized filtrate, taking a filter cake, rinsing with 10-20g of the ethanol solution II, pulping the rinsed filter cake with acetone, filtering, and carrying out vacuum drying at 50-55 ℃ for 20 +/-5 h until the water content is less than 4.5%, thus obtaining the final refined diammonium glycyrrhizinate product.
Further, the glycyrrhizin G2And glycyrrhizin H2The structure of the compound is obtained by analyzing diammonium glycyrrhizinate raw material medicine samples by using an HP L C-ESIMS (HP L C-ESIMS) analysis method, and the specific steps are as follows:
(1) dissolving diammonium glycyrrhizinate raw material drug samples in water to prepare 0.1-0.2mg/ml test solution, and analyzing by liquid chromatography, wherein the sample introduction amount is 10-12 mu L, the flow rate is 1.0-2.0ml/min, the detection wavelength is 248nm, impurity 1, impurity 2 and diammonium glycyrrhizinate are obtained, the peak of impurity 1 is 5.7min, the peak of impurity 2 is 11.7min, and the peak of diammonium glycyrrhizinate is 8.1 min;
adding a liquid chromatogram-mass spectrum combination instrument, wherein the mass spectrum scanning range is as follows: m/z is 100-1000, the detection wavelength is 248nm, and the peak emergence time of the impurity 1 and the impurity 2 is 5.71min and 12.07min respectively;
(2) analysis of the test result of the diammonium glycyrrhizinate bulk drug sample HP L C- (-) -ESIMS:
the impurity 1 is determined to be glycyrrhizin G through the analysis of mass spectrum test results2The chemical structural formula is as follows:
the impurity 2 is determined to be glycyrrhizin through mass spectrum test result analysisH2The chemical structural formula is shown as follows;
specifically, in the step (1), chromatographic conditions comprise that an analytical column using octadecylsilane chemically bonded silica as a stationary phase and a mobile phase are acetonitrile and 0.1% phosphoric acid solution, the volume ratio of the acetonitrile to the phosphoric acid solution is 38:62, the column temperature is 30 ℃ and mass spectrometry detection in the step (1) uses acetonitrile and 0.3% acetic acid solution, the volume ratio of the acetonitrile to the acetic acid solution is 38:62, and mass spectrometry conditions comprise that the flow rate of dry gas is 8L/min, the spray air pressure is 30psi, the collision voltage is 1.00V, the high voltage of a capillary is 3500V and the end plate voltage is-500V
Specifically, in the step a, adding 0.6-0.8 time of volume of the acetic acid solution I to the diammonium glycyrrhizinate crude product; in the step a, adding 3-4 times of volume of the acetic acid solution II to the diammonium glycyrrhizinate primary refined product; in the step a, the volume fraction of the acetic acid solution I is 90-98%, and the volume fraction of the acetic acid solution II is 80-95%.
Specifically, the weight of the activated carbon in the step a is 8-10% of the primary refined diammonium glycyrrhizinate product, and the activated carbon is pre-activated for 1-2 hours at 140 +/-10 ℃ before adding.
Specifically, the centrifugation time in the step a is 0.5-1 h, and the pulping time is 1-2 h.
Specifically, the volume fraction of the ethanol solution I in the step a is 90-98%, and the volume fraction of the ethanol solution II in the step b is 80-90%; in the step b, adding 4-5 times of the volume of the secondary refined diammonium glycyrrhizinate into the ethanol solution II.
Specifically, the weight of the activated carbon in the step b is 4-6% of the secondary refined diammonium glycyrrhizinate product, and the activated carbon is pre-activated for 1-2 hours at the temperature of 140 +/-10 ℃ before adding.
Specifically, the filter cake in the step b is pulped for 2-3h by using acetone, and the weight of the acetone is 2-3 times that of the filter cake.
Specifically, the vacuum drying temperature in the step b is 50-55 ℃.
Compared with the prior art, the invention has the beneficial effects that:
the present application is directed to the prior art, sweetThe research on related substances in diammonium oxalate bulk drugs and preparations only stays at the detection stage of single maximum impurity, secondary maximum impurity and total amount of impurities, the specific structure identification of triterpenoid impurities is reported and researched less, and no clear removal method is found for impurities, and the like2And glycyrrhizin H2And carrying out detailed reasoning analysis on the cracking way of the glycyrrhizin, and determining the glycyrrhizin G according to the structural property of impurities and experimental results2Is easily dissolved in acetic acid and can be effectively removed by refining; glycyrrhizin H2Is easy to dissolve in ethanol and can be effectively removed; effectively reduces G in diammonium glycyrrhizinate2And H2The content of (a). The method of the invention improves the purity of the product and is suitable for industrial large-scale production.
Drawings
FIG. 1 is a HP L C-DAD chromatogram of a diammonium glycyrrhizinate bulk drug;
FIG. 2 shows a basic peak ion current chromatogram of HP L C- (-) -ESIMS of diammonium glycyrrhizinate bulk drug and primary and secondary mass spectrograms of main components;
FIG. 3 shows the first and second mass spectra of (-) -ESIMS of peak 1-3 of diammonium glycyrrhizinate bulk drug;
FIG. 4 shows the first-order and second-order mass spectra of the (-) -ESIMS of HP L C- (-) -ESIMS of impurity components 1 and 2 in the diammonium glycyrrhizinate bulk drug.
Detailed Description
The technical solution of the present invention is further described in detail by the following specific examples, but the scope of the present invention is not limited thereto.
The diammonium glycyrrhizinate raw material medicine used in the method is a product self-made by medicinal industry GmbH of Henan Yuchen, the chemical name of the main component diammonium glycyrrhizinate is 20 β -carboxyl-11-oxo-n-oleanane-12-alkene-3 β -yl-2-O- β -D-glucopyranoside aldehydic acid- α -D-glucopyranoside aldehydic acid diammonium salt, and the structural formula is as follows:
example 1
The triterpenes impurities in diammonium glycyrrhizinate comprises glycyrrhizin G2And glycyrrhizin H2For glycyrrhizin G2And glycyrrhizin H2The structure of the method is obtained by analyzing diammonium glycyrrhizinate bulk drug samples by using an Agilent 1100S L liquid chromatography-mass spectrometer with HP L C-ESIMS, and the method comprises the following specific steps:
(1) dissolving a diammonium glycyrrhizinate raw material medicine sample with water to prepare a test solution of 0.2mg/ml, and analyzing by liquid chromatography, wherein the sample introduction amount is 10 mu L, the mobile phase is acetonitrile-0.1% phosphoric acid solution (38:: 62), the chromatographic column is ODS-C18 (octadecylsilane chemically bonded silica (250mm × 4.6.6 mm × 5 mu m)), the flow rate is 1.0ml/min, the detection wavelength is 248nm, the column temperature is 30 ℃, and under the chromatographic condition, impurity 1, impurity 2 and diammonium glycyrrhizinate are obtained, the peak of impurity 1 is 5.7min, the peak of impurity 2 is 11.7min, and the peak of diammonium glycyrrhizinate is 8.1 min;
adding a liquid chromatography-mass spectrometer, wherein a mobile phase used for mass spectrometry is acetonitrile and 0.3% acetic acid solution, the mass spectrometry scanning range is m/z 100-1000, the detection wavelength is 248nm, the flow rate of drying gas is 8L/min, the spraying air pressure is 30psi, the collision voltage is 1.00V, the capillary high voltage is 3500V, the end plate voltage is-500V, and under the condition, the peak emergence time of the impurity 1 and the impurity 2 is 5.71min and 12.07min respectively;
(2) analysis of the test result of the diammonium glycyrrhizinate bulk drug sample HP L C- (-) -ESIMS:
in HP L C- (-) -ESIMS, peak1, peak2 and peak3 in the primary mass spectrum respectively give excimer ions m/z837, m/z821 and m/z821, and 3 chromatographic peaks in the secondary mass spectrum give fragment ions m/z351, specifically:
the (-) -ESIMS of the impurity 1 gives an excimer ion peak m/z837 in a high-mass region, 16 mass units more than the excimer ion peak m/z821 of diammonium glycyrrhizinate as a main component in a sample, and the secondary mass spectrum of the impurity 1 gives a fragment ion m/z351, and the fragment ion also appears in the secondary mass spectrum of the glycyrrhizinate excimer ion m/z821 as the main component, which can be interpreted as a single-atom ion of diglucuronic acid generated by cracking the molecular ion and the loss of H2O and simultaneous loss of H2O and CO2M/z819 and 775 of (1), two ions m/z661 and m/z643 which are cleaved from different C-O bonds and lose one glucuronic acid fragment, and therefore, it is inferred that the structure of impurity 1 has the same diglucosic acid moiety as glycyrrhizic acid which is the main component, and the aglycone moiety thereof has one more hydroxyl group than glycyrrhizic acid, and is referred to and compared with the literature (Isao kitagawa, Junliang Zhou, Masahiro Sackagami, et al. L icornice-saponarins F3, G2, H2, J2 and K2.five new oleanone-triglucosides from the road of Glycyrrhiza Uraensis. chem. balance. 1991,39(1), 244; Isao tagagawa, Junliang hominix, Saccharomyces cerevisiae. m. chem. 31, 2, 9. Glycyrrhiza aethion 2. 3513. C9. 7. Glycyrrhiza aethion. 3532. 7. C9. 7. Glycyrrhiza. five saponin 1. 7. III. C9. 7. C9. five2(licorice-saponin G2)；
Glycyrrhizasaponin G2: the retention time is 5.71 min;
the (-) -ESIMS of the impurity 2 gives an excimer ion peak m/z821 in a high-quality area, which is the same as the excimer ion peak of the diammonium glycyrrhizinate serving as the main component; the impurity 2 has the same double glucuronic acid monoatomic ion m/z351 as the glycyrrhizic acid and the impurity 1 as the main components in the secondary mass spectrum, and the loss H is generated similarly to the glycyrrhizic acid2O and simultaneous loss of H2O and CO2The ion m/z803 and 759 of (1), and therefore, it is inferred that impurity 2 has the same diglucuronic acid moiety as glycyrrhizic acid and impurity 1 as the main components, and the aglycone moiety thereof is an isomer of glycyrrhetinic acid, which is referred to and found in the literature (Isao kitagawa, Junliang Zhou, Masahiro Sakagami, et al L icon-saponarins F3, G2, H2, J2 and K2.five new maize-triple oligosaccharide from the road of Glycyrrhiza Ugalensis Chem.Pharm.Bu1991, 39 (1); 244. 246.; Isao kitagawa, Junliang Zhou, Masahiro kagami, Ethyl L. Saponicins 2, Isao bagging 2, Junliang Zhou, Glycyrrhiza maize 3655, Glycyrrhiza maize 2, Glycyrrhiza maize 3655, Junlian maize 2, Glycyrrhiza maize 3645, Glycyrrhiza maize, III, Glycyrrhiza maize 80, III, Glycyrrhiza maize 2, Glycyrrhiza maize L, C2, Glycyrrhiza, C2, C3645rrhizaradix, chem, pharm, bull, 1988,36 (9); 3710-3713) and determining that the impurity 2 is glycyrrhizin H2(licorice-saponin H2)；
Glycyrrhizin H2: the retention time is 12.07 min;
according to the analysis result of HP L C- (-) -ESIMS, the impurity 1 glycyrrhizin G is subjected to2And impurity 2 glycyrrhizin H2The cleavage route is subjected to detailed reasoning analysis, and specifically comprises the following steps:
example 2
A method for removing triterpenes impurities in diammonium glycyrrhizinate raw material medicine, wherein the triterpenes impurities comprise glycyrrhizin G2And glycyrrhizin H2The method comprises the following specific steps:
a: glycyrrhizasaponin G2Removing, namely taking a diammonium glycyrrhizinate crude product, adding 95% (V/V) acetic acid solution with the volume of 0.6 times that of the diammonium glycyrrhizinate crude product, stirring, heating, fully dissolving, reacting at 90 ℃ for 30min, cooling to room temperature, standing, crystallizing for 72h, centrifuging for 0.5-1 h, taking a filter cake, washing the filter cake with 95% (V/V) acetic acid solution, centrifuging, filtering, weighing the filter cake, adding 95% (V/V) ethanol solution with the mass of 2 times that of the filter cake into the filter cake, pulping for 2h at room temperature, centrifuging for 0.5-1 h, washing the filter cake with 95% (V/V) ethanol solution, centrifuging, and drying in vacuum to obtain a diammonium glycyrrhizinate primary refined product; adding 87% (V/V) acetic acid solution 4 times of diammonium glycyrrhizinate into the primary refined productHeating and reacting for 20-30min at 96-97 ℃, stirring and fully dissolving, adding active carbon (the active carbon is pre-activated for 2h at 140 +/-10 ℃) which is 9.2 percent (W/W) of the weight of the primary refined diammonium glycyrrhizinate into the solution, keeping the temperature and decoloring for 20-30min, removing the carbon while hot and filtering, standing and crystallizing the filtrate for 72h at 0-10 ℃, centrifuging and filtering, rinsing with 87 percent (V/V) acetic acid solution, filtering to obtain a filter cake, and vacuum drying to obtain the secondary refined diammonium glycyrrhizinate;
b: glycyrrhizin H2Adding 85% (V/V) ethanol solution with the weight 5 times that of the diammonium glycyrrhizinate secondary refined product in the step b into a 250ml four-mouth reaction bottle, stirring to dissolve, heating to 85 ℃, adding 15.5g of the diammonium glycyrrhizinate secondary refined product in the step b in batches, continuously heating to 78 ℃ until the solution is clear, adding 0.8g of active carbon (the weight of the active carbon is 4.8% (W/W) of the diammonium glycyrrhizinate secondary refined product) at the temperature of 78 +/-3 ℃ until the solution is clear, pre-activating at 140 +/-10 ℃ for 2 hours before adding, refluxing and decoloring at 78 +/-3 ℃ for 20min, filtering while hot, cooling the filtrate, refrigerating and crystallizing for 3 days, filtering the crystallized filtrate, rinsing the filter cake with 10-20g of cold 85% (V/V) ethanol solution, pulping the obtained filter cake with acetone (the weight 2 times that of the filter cake) for 2 hours, filtering, vacuum drying at 50 deg.C for 20 + -5 hr until water content is less than 4.5%, to obtain refined diammonium glycyrrhizinate product with yield not less than 50%.
Through the steps, the diammonium glycyrrhizinate crude products are refined, the unrefined crude products are refined for the first time and refined for the second time through the steps, and the detection comparison results of triterpenoid impurities in diammonium glycyrrhizinate bulk drugs in commercially available products are summarized as the following table 1:
TABLE 1
Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A method for removing triterpenes impurities in diammonium glycyrrhizinate bulk drug is characterized in that the triterpenes impurities comprise glycyrrhizin G2And glycyrrhizin H2The removing method comprises the following steps:
a: glycyrrhizasaponin G2Removing, namely adding acetic acid solution I into a diammonium glycyrrhizinate crude product, heating to 85-95 ℃, reacting for 25-35min, cooling to room temperature, standing to separate out crystals for 66-72h, centrifuging, taking a filter cake, washing the filter cake with the acetic acid solution I, centrifuging, draining, adding ethanol solution I with 2-3 times of the mass of the filter cake, pulping at room temperature, centrifuging, washing the filter cake with the ethanol solution I, centrifuging, draining the filter cake, and drying in vacuum to obtain a diammonium glycyrrhizinate primary refined product; adding an acetic acid solution II which is 3-5 times of the weight of the primary refined diammonium glycyrrhizinate into the primary refined diammonium glycyrrhizinate, heating to 96-97 ℃, reacting for 20-30min, adding active carbon, carrying out heat preservation and decoloration for 20-30min, removing carbon while hot, filtering, standing filtrate at 0-10 ℃ for crystallization for 66-72h, carrying out centrifugal filtration, rinsing with the acetic acid solution II, and carrying out vacuum drying on a filter cake to obtain a secondary refined diammonium glycyrrhizinate;
b: glycyrrhizin H2Removing, namely heating the ethanol solution II to 85-90 ℃, adding 15-18g of the diammonium glycyrrhizinate secondary refined product obtained in the step a, continuously heating to 75-81 ℃, adding 0.6-1.0g of active carbon at the temperature of 78 +/-3 ℃, carrying out reflux decolorization for 10-20min at the temperature of 78 +/-3 ℃, filtering, cooling the filtrate, refrigerating and crystallizing for 3-4d, filtering the crystallized filtrate, taking a filter cake, rinsing with 10-20g of the ethanol solution II, pulping the filter cake with acetone after rinsing, filtering, and carrying out vacuum drying for 20 +/-5 h until the water content is less than 4.5%, thus obtaining the final refined diammonium glycyrrhizinate product.
2. The removal method as claimed in claim 1, wherein the licorice soap isGlycoside G2And glycyrrhizin H2The structure of the compound is obtained by using an HP L C-ESI/MS analysis method for diammonium glycyrrhizinate raw material medicine samples, and the specific steps are as follows:
(1) dissolving diammonium glycyrrhizinate raw material drug samples in water to prepare 0.1-0.2mg/ml test solution, and analyzing by liquid chromatography, wherein the sample introduction amount is 10-12 mu L, the flow rate is 1.0-2.0ml/min, the detection wavelength is 248nm, impurity 1, impurity 2 and diammonium glycyrrhizinate are obtained, the peak of impurity 1 is 5.7min, the peak of impurity 2 is 11.7min, and the peak of diammonium glycyrrhizinate is 8.1 min;
(2) adding a liquid chromatogram-mass spectrum combination instrument, wherein the mass spectrum scanning range is as follows: m/z is 100-1000, the detection wavelength is 248nm, and the peak emergence time of the impurity 1 and the impurity 2 is 5.71min and 12.07min respectively; the impurity 1 is determined to be glycyrrhizin G through the analysis of mass spectrum test results2The chemical structural formula is as follows:
determining impurity 2 as glycyrrhizin H2The chemical structural formula is shown as follows;
3. the removing method according to claim 2, wherein the chromatographic conditions in step (1) are an analytical column using octadecylsilane chemically bonded silica as a stationary phase, the mobile phase is acetonitrile: 0.1% phosphoric acid solution, the volume ratio of the acetonitrile: 0.1% phosphoric acid solution and the phosphoric acid solution is 38:62, the column temperature is 30 ℃, the mobile phase used in the mass spectrometry in step (1) is acetonitrile: 0.3% acetic acid solution, the volume ratio of the acetonitrile: 0.62, the mass spectrometry conditions are that the drying gas flow rate is 8L/min, the spray gas pressure is 30psi, the collision voltage is 1.00V, the capillary high voltage is 3500V, and the end plate voltage is-500V.
4. The removing method according to claim 1, wherein the volume of the acetic acid solution I added in step a is 0.6-0.8 times of the volume of the crude diammonium glycyrrhizinate; in the step a, adding 3-4 times of volume of the acetic acid solution II to the diammonium glycyrrhizinate primary refined product; in the step a, the volume fraction of the acetic acid solution I is 90-98%, and the volume fraction of the acetic acid solution II is 80-95%.
5. The removing method according to claim 1, wherein the weight of the activated carbon in the step a is 8-10% of the primary refined diammonium glycyrrhizinate product, and the activated carbon is pre-activated for 1-2 hours at 140 ± 10 ℃ before being added.
6. The removing method according to claim 1, wherein the centrifugation time in step a is 0.5-1 h, and the beating time is 1-2 h.
7. The removal method according to claim 1, wherein the volume fraction of the ethanol solution I in the step a is 90-98%, and the volume fraction of the ethanol solution II in the step b is 80-90%; in the step b, adding 4-5 times of the volume of the secondary refined diammonium glycyrrhizinate into the ethanol solution II.
8. The removing method according to claim 1, wherein the weight of the activated carbon in the step b is 4-6% of the secondary refined diammonium glycyrrhizinate product, and the activated carbon is pre-activated for 1-2 hours at 140 ± 10 ℃ before being added.
9. The removing method according to claim 1, wherein the filter cake in the step b is beaten with acetone for 2 to 3 hours, the weight of acetone being 2 to 3 times that of the filter cake.
10. The removing method according to claim 1, wherein the vacuum drying temperature in the step b is 50 to 55 ℃.
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| CN114409720A (en) * | 2021-12-31 | 2022-04-29 | 江苏天晟药业股份有限公司 | Method for improving large production content of diammonium glycyrrhizinate |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102286058A (en) * | 2011-09-07 | 2011-12-21 | 西安瑞联近代电子材料有限责任公司 | Method for preparing glycyrrhizic acid with high purity |
| CN104861029A (en) * | 2015-01-30 | 2015-08-26 | 李玉山 | Preparation method of ammonium 18alpha,beta-H-glycyrrhetate and hydrate thereof |
| CN106380506A (en) * | 2016-08-29 | 2017-02-08 | 江苏天晟药业股份有限公司 | Preparation method of 18 alpha type diammonium glycyrrhizinate |
| CN106478762A (en) * | 2016-08-29 | 2017-03-08 | 江苏天晟药业股份有限公司 | A kind of preparation method of diammonium glycyrhetate |
| CN109111495A (en) * | 2018-05-02 | 2019-01-01 | 安徽大学 | A kind of method of related substances content in reduction mono-ammonium glycyrrhizinate |
| CN109232707A (en) * | 2018-09-17 | 2019-01-18 | 河南慧锦药业有限公司 | A kind of minimizing technology of diammonium glycyrrhizinate bulk pharmaceutical chemicals dissolvent residual |
-
2020
- 2020-03-25 CN CN202010215860.2A patent/CN111393499A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102286058A (en) * | 2011-09-07 | 2011-12-21 | 西安瑞联近代电子材料有限责任公司 | Method for preparing glycyrrhizic acid with high purity |
| CN104861029A (en) * | 2015-01-30 | 2015-08-26 | 李玉山 | Preparation method of ammonium 18alpha,beta-H-glycyrrhetate and hydrate thereof |
| CN106380506A (en) * | 2016-08-29 | 2017-02-08 | 江苏天晟药业股份有限公司 | Preparation method of 18 alpha type diammonium glycyrrhizinate |
| CN106478762A (en) * | 2016-08-29 | 2017-03-08 | 江苏天晟药业股份有限公司 | A kind of preparation method of diammonium glycyrhetate |
| CN109111495A (en) * | 2018-05-02 | 2019-01-01 | 安徽大学 | A kind of method of related substances content in reduction mono-ammonium glycyrrhizinate |
| CN109232707A (en) * | 2018-09-17 | 2019-01-18 | 河南慧锦药业有限公司 | A kind of minimizing technology of diammonium glycyrrhizinate bulk pharmaceutical chemicals dissolvent residual |
Non-Patent Citations (3)
| Title |
|---|
| 吕昭云等: "HPLC法分离18α-、18β-甘草酸并用于甘草酸类产品的质量控制", 《药物分析杂志》 * |
| 姜新等: "液相色谱-质谱联用技术鉴定甘草酸二铵肠溶片中有关物质", 《中草药》 * |
| 黄浩等: "我国甘草酸铵盐原料药质量控制方法的研究探讨", 《中成药》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114409720A (en) * | 2021-12-31 | 2022-04-29 | 江苏天晟药业股份有限公司 | Method for improving large production content of diammonium glycyrrhizinate |
| CN114409720B (en) * | 2021-12-31 | 2023-12-29 | 江苏天晟药业股份有限公司 | Method for improving large-scale production content of diammonium glycyrrhizinate |
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