CN111705097A - Purification process of radix astragali for animal feed additive - Google Patents
Purification process of radix astragali for animal feed additive Download PDFInfo
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- CN111705097A CN111705097A CN202010621104.XA CN202010621104A CN111705097A CN 111705097 A CN111705097 A CN 111705097A CN 202010621104 A CN202010621104 A CN 202010621104A CN 111705097 A CN111705097 A CN 111705097A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/44—Preparation of O-glycosides, e.g. glucosides
- C12P19/60—Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/14—Pretreatment of feeding-stuffs with enzymes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J53/00—Steroids in which the cyclopenta(a)hydrophenanthrene skeleton has been modified by condensation with a carbocyclic rings or by formation of an additional ring by means of a direct link between two ring carbon atoms, including carboxyclic rings fused to the cyclopenta(a)hydrophenanthrene skeleton are included in this class
- C07J53/002—Carbocyclic rings fused
- C07J53/004—3 membered carbocyclic rings
Abstract
The invention discloses a purification process of astragalus mongholicus for an animal feed additive, which comprises the following steps: pulverizing radix astragali, sieving, adding zeolite powder, grinding in a grinder, adding enzyme preparation, and treating at 30-35 deg.C for 30-50 hr to obtain a prefabricated material; adding anhydrous ethanol into the prefabricated material, heating and reflux-extracting for 2-3 times, mixing filtrates, concentrating under reduced pressure until the solid content is 10-20%, adding into sodium hydroxide solution, and reflux-stirring at 50-60 deg.C for 2-4 hr to obtain pre-extract; loading AB-8 resin into a chromatographic column, and loading the pre-extraction solution; washing with distilled water to remove sugar; eluting with sodium hydroxide solution to remove impurities, and washing with distilled water to neutrality; eluting with ethanol solution at an elution flow rate of 210-220L/h, and collecting the eluate to obtain purified solution; removing ethanol in the purified solution, filtering, adding the filter cake into 90-95% ethanol by mass percent, and recrystallizing to obtain the purified baicalin.
Description
Technical Field
The invention relates to the technical field of feed additives, in particular to a purification process of astragalus mongholicus for an animal feed additive.
Background
Astragalus membranaceus is a plant belonging to the genus Astragalus of the family Leguminosae, a perennial herb, and has a thick main root, is woody, frequently branched, off-white, upright stem, multi-branched upper part, fine edges and white and soft hair. The feathery compound leaves have 13-27 small leaves and are 5-10 cm long. The general inflorescence of the astragalus is slightly dense, and 10-20 flowers exist; the total pedicel is nearly as long as the leaf or longer until the fruit stage is obviously elongated. The pod is membranous, slightly swollen, semi-elliptical, with the neck of the fruit beyond the calyx; 3-8 seeds. The flowering period is 6-8 months, and the fruit period is 7-9 months.
The astragalus root can be used as a medicine, has sweet taste and mild nature, and pharmacological research shows that the astragalus contains choline, coumarin, folic acid, amino acid, betaine, saponin, saccharide, protein, riboflavin, flavan compound, iron, calcium, phosphorus, selenium, zinc, copper, manganese and other trace elements. The astragalus membranaceus has the effects of tonifying qi, strengthening exterior, promoting urination, strengthening heart, reducing blood pressure, resisting bacteria, expelling toxin, expelling pus, promoting granulation, strengthening capillary resistance, suppressing sweating and sex hormones, and is used for treating exterior deficiency spontaneous perspiration, internal injury due to qi deficiency, spleen deficiency diarrhea, edema, carbuncle and cellulitis and the like.
Researches show that the saponin compound is one of the important components of the drug effect substance basis of the astragalus, and the astragaloside is one of the main active ingredients of the astragalus. Astragaloside IV is a kind of lanolate alcohol type tetracyclic triterpenoid saponin, and is an important index component for evaluating the astragalus root medicinal material and the preparation containing the astragalus root medicinal material at present. However, the content of astragaloside in astragalus is very low, about 0.04%, extraction, separation and purification are difficult, and the yield and purity of astragaloside are low, so that a solution is needed.
Disclosure of Invention
The invention aims to solve the defects in the prior art and provides a process for purifying astragalus membranaceus used for an animal feed additive.
A purification process of radix astragali for animal feed additive comprises the following steps:
s1, crushing and sieving the astragalus, adding zeolite powder, feeding the mixture into a grinder for grinding for 10-20min, wherein the grinding pressure is 1.25-1.45MPa, the temperature is less than or equal to 50 ℃ in the grinding process, then adding an enzyme preparation, and treating for 30-50h at 30-35 ℃ to obtain a prefabricated material;
s2, adding absolute ethyl alcohol into the prefabricated material, soaking for 10-20h at 40-50 ℃, then heating and refluxing for 2-3 times, combining filtrates, concentrating under reduced pressure until the solid content is 10-20%, adding into a sodium hydroxide solution, and refluxing and stirring for 2-4h at 50-60 ℃ to obtain a pre-extraction solution;
s3, loading the AB-8 resin into a chromatographic column, and loading the pre-extraction solution at an adsorption flow rate of 100-; removing sugar by using distilled water, wherein the elution flow rate is 200-210L/h; eluting with 0.12-0.16mol/L sodium hydroxide solution at an elution flow rate of 200-210L/h to remove impurities, and washing with distilled water to neutrality; eluting with 55-65% ethanol solution at an elution flow rate of 210-220L/h, and collecting eluate to obtain purified solution;
s4, removing ethanol in the purified liquid, filtering, and adding the filter cake into ethanol with the mass fraction of 90-95% for recrystallization to obtain the purified baicalin.
Preferably, in S1, the mass ratio of the astragalus to the zeolite powder to the enzyme preparation is 10-20: 2-4: 50-80.
Preferably, in S1, the polishing speed is 1000-1200 r/min.
Preferably, the enzyme preparation used in S1 has a concentration of 0.1-0.12u/ml of deacetylase, a concentration of 0.15-0.2u/ml of glycosidase and a concentration of 0.08-0.12u/ml of cellulase.
Preferably, in S2, absolute ethyl alcohol is added into the prefabricated material until the mass fraction of the ethyl alcohol in the system is 60-70%.
Preferably, in S2, the concentration of the sodium hydroxide solution is 2-3mol/L, and the mass ratio of the preformed material to the sodium hydroxide solution is 100: 20-30.
Preferably, the yield of astragaloside obtained from S4 is more than 0.079%, and the purity is more than 99.4%.
The technical effects of the invention are as follows:
according to the invention, astragalus is firstly crushed and then compounded and ground with zeolite powder, by controlling grinding parameters, the dissolution degree of active ingredients is extremely high on the premise of not damaging the active ingredients of the astragalus, and the active ingredients are matched with an enzyme preparation for treatment, and the enzyme preparation is fully activated in a system, so that the dissolution degree of astragaloside can be effectively improved, and other dissolved astragalosides can be converted into astragaloside, and the method has the advantages of mild reaction conditions, simplicity and convenience in operation, less pollution, capability of effectively controlling the problem of instability in an enzymatic process, extremely high product specificity and extremely high conversion rate;
the method is matched with AB-8 resin chromatography treatment, so that the content of impurities in the pre-extraction solution is reduced, the extraction cost is reduced by avoiding using various filtering membranes, the problems of low dissolution degree and serious loss of the astragaloside can be solved, and the high-yield and high-purity astragaloside can be obtained simply and quickly.
Detailed Description
The present invention will be further illustrated with reference to the following specific examples.
Example 1
A purification process of radix astragali for animal feed additive comprises the following steps:
s1, crushing 10kg of astragalus, sieving with a 80-mesh sieve, adding 4kg of zeolite powder, feeding into a grinder for grinding for 10min at the grinding speed of 1200r/min and the grinding pressure of 1.25MPa, maintaining the temperature at 50 ℃ in the grinding process, then adding 50kg of enzyme preparation, and treating at 35 ℃ for 30h to obtain a prefabricated material;
in the enzyme preparation, the concentration of the deacetylase is 0.12u/ml, the concentration of the glycosidase is 0.15u/ml, and the concentration of the cellulase is 0.12 u/ml;
s2, adding absolute ethyl alcohol into 100kg of prefabricated material until the mass fraction of the ethyl alcohol in the system is 60%, soaking for 10h at 50 ℃, then heating and refluxing for 3 times, combining filtrates, concentrating under reduced pressure until the solid content is 10%, adding into 30kg of sodium hydroxide solution with the concentration of 2mol/L, and refluxing and stirring for 2h at 60 ℃ to obtain a pre-extracting solution;
s3, filling the AB-8 resin into a chromatographic column, and loading the pre-extraction solution into the chromatographic column, wherein the adsorption flow rate is 106L/h and the pre-extraction solution is saturated for 2 h; washing with distilled water to remove sugar, wherein the elution flow rate is 200L/h; eluting with 0.16mol/L sodium hydroxide solution at 200L/h to remove impurities, and washing with distilled water to neutrality; eluting with 65% ethanol solution at an elution flow rate of 210L/h, and collecting eluate to obtain purified solution;
s4, removing ethanol in the purified liquid, filtering, adding the filter cake into 95% ethanol by mass percent, and recrystallizing to obtain purified baicalin;
the yield of astragaloside obtained in this example was 0.079%, and the purity was 99.4%.
Example 2
A purification process of radix astragali for animal feed additive comprises the following steps:
s1, crushing 20kg of astragalus membranaceus, sieving the crushed astragalus membranaceus with an 80-mesh sieve, adding 2kg of zeolite powder, feeding the mixture into a grinder to grind for 20min at the grinding speed of 1000r/min and the grinding pressure of 1.45MPa, maintaining the temperature at 45 ℃ in the grinding process, then adding 80kg of enzyme preparation, and treating the mixture at 30 ℃ for 50h to obtain a prefabricated material;
in the enzyme preparation, the concentration of the deacetylase is 0.1u/ml, the concentration of the glycosidase is 0.2u/ml, and the concentration of the cellulase is 0.08 u/ml;
s2, adding absolute ethyl alcohol into 100kg of prefabricated material until the mass fraction of the ethyl alcohol in the system is 70%, soaking for 20h at 40 ℃, then heating and refluxing for 2 times, combining filtrates, concentrating under reduced pressure until the solid content is 20%, adding into 20kg of sodium hydroxide solution with the concentration of 3mol/L, and refluxing and stirring for 4h at 50 ℃ to obtain a pre-extracting solution;
s3, filling the AB-8 resin into a chromatographic column, and loading the pre-extraction solution into the chromatographic column, wherein the adsorption flow rate is 100L/h and the pre-extraction solution is saturated for 2 h; washing with distilled water to remove sugar, wherein the elution flow rate is 210L/h; eluting with 0.12mol/L sodium hydroxide solution at flow rate of 210L/h to remove impurities, and washing with distilled water to neutrality; eluting with 55% ethanol solution at an elution flow rate of 220L/h, and collecting eluate to obtain purified solution;
s4, removing ethanol in the purified liquid, filtering, adding the filter cake into ethanol with the mass fraction of 90%, and recrystallizing to obtain purified baicalin;
the yield of astragaloside obtained in this example was 0.081%, and the purity was 99.5%.
Example 3
A purification process of radix astragali for animal feed additive comprises the following steps:
s1, crushing 12kg of astragalus, sieving with a 80-mesh sieve, adding 3.5kg of zeolite powder, grinding for 13min in a grinder at the grinding speed of 1150r/min and the grinding pressure of 1.3MPa, maintaining the temperature at 46 ℃ in the grinding process, adding 60kg of enzyme preparation, and treating for 35h at 34 ℃ to obtain a prefabricated material;
in the enzyme preparation, the concentration of the deacetylase is 0.115u/ml, the concentration of the glycosidase is 0.16u/ml, and the concentration of the cellulase is 0.11 u/ml;
s2, adding absolute ethyl alcohol into 100kg of prefabricated material until the mass fraction of the ethyl alcohol in the system is 63%, soaking for 13h at 48 ℃, then heating and refluxing for 3 times, combining filtrates, concentrating under reduced pressure until the solid content is 13%, adding into 28kg of 2.2mol/L sodium hydroxide solution, and refluxing and stirring for 2.5h at 56 ℃ to obtain a pre-extracting solution;
s3, filling the AB-8 resin into a chromatographic column, and loading the pre-extraction solution into the chromatographic column, wherein the adsorption flow rate is 104L/h and the pre-extraction solution is saturated for 2 h; washing with distilled water to remove sugar, wherein the elution flow rate is 200L/h; eluting with 0.15mol/L sodium hydroxide solution at flow rate of 202L/h to remove impurities, and washing with distilled water to neutrality; eluting with 62% ethanol solution at an elution flow rate of 213L/h, and collecting eluate to obtain purified solution;
s4, removing ethanol in the purified liquid, filtering, adding the filter cake into ethanol with the mass fraction of 94% for recrystallization to obtain purified baicalin;
the yield of astragaloside obtained in this example was 0.082%, and the purity was 99.8%.
Example 4
A purification process of radix astragali for animal feed additive comprises the following steps:
s1, crushing 18kg of astragalus, sieving with a 80-mesh sieve, adding 2.5kg of zeolite powder, grinding for 17min in a grinder at the grinding speed of 1050r/min and the grinding pressure of 1.4MPa, maintaining the temperature at 40 ℃ in the grinding process, adding 70kg of enzyme preparation, and treating for 45h at 32 ℃ to obtain a prefabricated material;
in the enzyme preparation, the concentration of the deacetylase is 0.105u/ml, the concentration of the glycosidase is 0.18u/ml, and the concentration of the cellulase is 0.09 u/ml;
s2, adding absolute ethyl alcohol into 100kg of prefabricated material until the mass fraction of the ethyl alcohol in the system is 67%, soaking for 17h at 42 ℃, then heating and refluxing for 2 times, combining filtrates, concentrating under reduced pressure until the solid content is 17%, adding into 22kg of sodium hydroxide solution with the concentration of 2.8mol/L, and refluxing and stirring for 3.5h at 54 ℃ to obtain a pre-extracting solution;
s3, filling the AB-8 resin into a chromatographic column, and loading the pre-extraction solution into the chromatographic column, wherein the adsorption flow rate is 102L/h and the pre-extraction solution is saturated for 2 h; washing with distilled water to remove sugar, wherein the elution flow rate is 210L/h; eluting with 0.13mol/L sodium hydroxide solution at a flow rate of 208L/h to remove impurities, and washing with distilled water to neutrality; eluting with 58% ethanol solution at an elution flow rate of 217L/h, and collecting eluate to obtain purified solution;
s4, removing ethanol in the purified liquid, filtering, adding the filter cake into 92% ethanol by mass percent, and recrystallizing to obtain purified baicalin;
the yield of astragaloside obtained in this example was 0.084% and the purity was 99.7%.
Example 5
A purification process of radix astragali for animal feed additive comprises the following steps:
s1, crushing 15kg of astragalus membranaceus, sieving the crushed astragalus membranaceus with a 80-mesh sieve, adding 3kg of zeolite powder, feeding the mixture into a grinder to grind for 13min, wherein the grinding speed is 1100r/min, the grinding pressure is 1.35MPa, the temperature is maintained at 43 ℃ in the grinding process, then adding 65kg of enzyme preparation, and treating the mixture at 33 ℃ for 40h to obtain a prefabricated material;
in the enzyme preparation, the concentration of the deacetylase is 0.11u/ml, the concentration of the glycosidase is 0.17u/ml, and the concentration of the cellulase is 0.1 u/ml;
s2, adding absolute ethyl alcohol into 100kg of prefabricated material until the mass fraction of the ethyl alcohol in the system is 65%, soaking for 15h at 45 ℃, then heating and refluxing for 3 times, combining filtrates, concentrating under reduced pressure until the solid content is 15%, adding 25kg of sodium hydroxide solution with the concentration of 2.5mol/L, and refluxing and stirring for 3h at 55 ℃ to obtain a pre-extracting solution;
s3, filling the AB-8 resin into a chromatographic column, and loading the pre-extraction solution into the chromatographic column, wherein the adsorption flow rate is 103L/h and the pre-extraction solution is saturated for 2 h; washing with distilled water to remove sugar, wherein the elution flow rate is 205L/h; eluting with 0.14mol/L sodium hydroxide solution at flow rate of 205L/h to remove impurities, and washing with distilled water to neutrality; eluting with 60% ethanol solution at an elution flow rate of 215L/h, and collecting the eluate to obtain a purified solution;
s4, removing ethanol in the purified liquid, filtering, adding a filter cake into 93% ethanol by mass percent, and recrystallizing to obtain purified baicalin;
the yield of astragaloside obtained in this example was 0.085% and the purity was 99.9%.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (7)
1. A purification process of radix astragali for an animal feed additive is characterized by comprising the following steps:
s1, crushing and sieving the astragalus, adding zeolite powder, feeding the mixture into a grinder for grinding for 10-20min, wherein the grinding pressure is 1.25-1.45MPa, the temperature is less than or equal to 50 ℃ in the grinding process, then adding an enzyme preparation, and treating for 30-50h at 30-35 ℃ to obtain a prefabricated material;
s2, adding absolute ethyl alcohol into the prefabricated material, soaking for 10-20h at 40-50 ℃, then heating and refluxing for 2-3 times, combining filtrates, concentrating under reduced pressure until the solid content is 10-20%, adding into a sodium hydroxide solution, and refluxing and stirring for 2-4h at 50-60 ℃ to obtain a pre-extraction solution;
s3, loading the AB-8 resin into a chromatographic column, and loading the pre-extraction solution at an adsorption flow rate of 100-; removing sugar by using distilled water, wherein the elution flow rate is 200-210L/h; eluting with 0.12-0.16mol/L sodium hydroxide solution at an elution flow rate of 200-210L/h to remove impurities, and washing with distilled water to neutrality; eluting with 55-65% ethanol solution at an elution flow rate of 210-220L/h, and collecting eluate to obtain purified solution;
s4, removing ethanol in the purified liquid, filtering, and adding the filter cake into ethanol with the mass fraction of 90-95% for recrystallization to obtain the purified baicalin.
2. The process for purifying astragalus membranaceus as animal feed additive according to claim 1, wherein in S1, the mass ratio of astragalus membranaceus to zeolite powder to enzyme preparation is 10-20: 2-4: 50-80.
3. The process for purifying astragalus membranaceus as claimed in claim 1, wherein the grinding speed of S1 is 1000-.
4. The process for purifying astragalus membranaceus as claimed in claim 1, wherein the concentration of deacetylase in the enzyme preparation used in S1 is 0.1-0.12u/ml, the concentration of glycosidase in the enzyme preparation used in S1 is 0.15-0.2u/ml, and the concentration of cellulase in the enzyme preparation used in S1 is 0.08-0.12 u/ml.
5. The process for purifying astragalus membranaceus as claimed in claim 1, wherein anhydrous ethanol is added to the preformed material in the step S2 until the mass fraction of ethanol in the system is 60-70%.
6. The process for purifying astragalus membranaceus as an animal feed additive according to claim 1, wherein in S2, the concentration of the sodium hydroxide solution is 2-3mol/L, and the mass ratio of the prefabricated material to the sodium hydroxide solution is 100: 20-30.
7. The process for purifying astragalus membranaceus as claimed in claim 1, wherein the yield of astragaloside obtained from S4 is 0.079% or more, and the purity is 99.4% or more.
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Citations (5)
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|---|---|---|---|---|
| CN101775056A (en) * | 2010-01-28 | 2010-07-14 | 东北林业大学 | Method for extracting, separating and purifying Astragaloside IV from Astragalus mongholicus |
| CN102559828A (en) * | 2010-12-30 | 2012-07-11 | 复旦大学 | Method for preparing astragaloside IV by converting total saponins of astragalus by microorganisms |
| CN102746362A (en) * | 2011-04-19 | 2012-10-24 | 河北以岭医药研究院有限公司 | Method for extracting refined astragaloside from astragaliradix |
| CN103073614A (en) * | 2013-01-22 | 2013-05-01 | 西安岳达植物科技有限公司 | Method for extracting high-purity astragaloside from astragalus mongholicus |
| CN110669096A (en) * | 2019-11-15 | 2020-01-10 | 齐齐哈尔医学院 | Method for preparing astragaloside from astragalus |
-
2020
- 2020-06-30 CN CN202010621104.XA patent/CN111705097A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101775056A (en) * | 2010-01-28 | 2010-07-14 | 东北林业大学 | Method for extracting, separating and purifying Astragaloside IV from Astragalus mongholicus |
| CN102559828A (en) * | 2010-12-30 | 2012-07-11 | 复旦大学 | Method for preparing astragaloside IV by converting total saponins of astragalus by microorganisms |
| CN102746362A (en) * | 2011-04-19 | 2012-10-24 | 河北以岭医药研究院有限公司 | Method for extracting refined astragaloside from astragaliradix |
| CN103073614A (en) * | 2013-01-22 | 2013-05-01 | 西安岳达植物科技有限公司 | Method for extracting high-purity astragaloside from astragalus mongholicus |
| CN110669096A (en) * | 2019-11-15 | 2020-01-10 | 齐齐哈尔医学院 | Method for preparing astragaloside from astragalus |
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| YU KONG ET AL: "Preparative enrichment and separation of astragalosides from Radix Astragali extracts using macroporous resins", 《LIQUID CHROMATOGRAPHY》 * |
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