CN111321179A - Method for preparing matrine from subprostrate sophora - Google Patents
Method for preparing matrine from subprostrate sophora Download PDFInfo
- Publication number
- CN111321179A CN111321179A CN201811536654.0A CN201811536654A CN111321179A CN 111321179 A CN111321179 A CN 111321179A CN 201811536654 A CN201811536654 A CN 201811536654A CN 111321179 A CN111321179 A CN 111321179A
- Authority
- CN
- China
- Prior art keywords
- filtrate
- ethanol solution
- filtering
- matrine
- concentrating
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- 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
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/18—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms containing at least two hetero rings condensed among themselves or condensed with a common carbocyclic ring system, e.g. rifamycin
- C12P17/182—Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides a method for preparing matrine from subprostrate sophora root, which comprises the following steps: crushing the vietnamese sophora root, adding an ethanol solution, leaching for 1-3 hours at a high temperature of 80-90 ℃, and filtering to obtain filtrate I and filter residue I; adding 5-10% of an Aspergillus niger liquid chamber into the filter residue I, carrying out warm fermentation for 3-8 days, adding an ethanol solution, carrying out high-temperature leaching at 80-90 ℃ for 1-3 h, and filtering to obtain a filtrate II; mixing the filtrate I and the filtrate II, concentrating under reduced pressure, and extracting with ethyl acetate with a volume twice to four times that of the filtrate I to obtain a water phase; adjusting the pH value of the water phase to 7.5-8, and extracting with butanol with a volume which is two to four times that of the water phase to obtain a butanol phase; concentrating butanol phase, loading on ADS-7 column, eluting with ethanol solution four times to six times column volume to obtain eluate I; and concentrating the eluent I, crystallizing overnight at 4-8 ℃, and filtering to obtain a matrine product.
Description
Technical Field
The invention relates to a method for preparing matrine from subprostrate sophora tonkinensis, belonging to the technical field of natural product preparation.
Background
The radix Sophorae Tonkinensis is a plant belonging to the genus Vietnamese of the family Leguminosae, and is also named as Vietnamese Asiatic moonseed root, Vietnamese sophora root, and rhizoma Schizocapsae Plantagineae (in the western Hunan province). The plants of Sophora contain alkaloids, flavonoids, steroids, and volatile oil. The plants in the genus contain various flavonoid compounds, and 86 flavonoid glycosides are obtained from the plants in the genus at present, wherein one flavonoid glycoside is flavonoid aglycone, and the rest flavonoid aglycone is flavonoid aglycone. 13 flavonoids are separated from roots of subprostrate sophora root, including 6 subprostrata (euchretin A-E, I) and 7 subprostrata ketones (euchrenone a1, a4, a14, a15, b2, b16, b 17). The alkaloid separated from the plant of the genus is 14, and plum exists and the like, which are separated from the subprostrate sophora calyx to obtain matrine, oxymatrine and cytisine, but are only identified by chemical and infrared spectroscopy analysis, and a more precise nuclear magnetic resonance method (NMR) is not used. And others: comprises 4 steroids, 40 volatile oils and coumarins thereof, wherein coumarone benzofurans are obtained by separating subprostrate sophora calyx. The medicinal value of the subprostrate sophora plants is widely reputed at home and abroad, and the subprostrate sophora is used for treating ulcer, diminishing inflammation, resisting cancer and resisting arrhythmia in Japan; in java island, seeds of Euchresta horsfieldii were used as kidney tonics. The tonkin sophora root is known as a national medicine with extremely high reputation in the Tujia mountain area of China, and the plant has strong effects of clearing heat, relieving pain, diminishing inflammation, resisting cancer and the like and has good curative effects on sore throat, toothache, stomachache, poisonous insect bite and the like. In addition, the Zhu imperial and other reports that the subprostrate sophora root is used as a main drug to prepare the burn liniment which is clinically applied and has satisfactory effect. Pharmacological research shows that the plant has the activities of resisting tumor, oxidation, bacteria, platelet aggregation, HIV, central depression, blood fat regulation, etc.
Chinese patent publication No. CN103630630A discloses a method for determining the content of matrine in subprostrate sophora root, which is characterized in that: (1) octadecylsilane chemically bonded silica is used as a filler in chromatographic conditions and system applicability tests; isopropanol, acetonitrile, 0.1% phosphoric acid (74:18:8) are used as a mobile phase; the detection wavelength is 210 nm; the number of theoretical plates is not less than 2000 calculated according to matrine peak; (2) preparing reference solution by accurately weighing appropriate amount of matrine reference, and adding mobile phase to obtain solution containing matrine 20 μ g per 1 ml; (3) preparing a test solution, namely taking about 0.5g of powder of the test solution, sieving the powder by a third sieve, precisely weighing the powder, placing the powder into a conical flask with a plug, precisely adding 50% ethanol 50ml, sealing the plug, weighing the powder, placing the powder for 30 minutes, carrying out ultrasonic treatment for 30 minutes, carrying out power of 250W and frequency of 40kHz, weighing the powder again, supplementing the lost weight by 50% ethanol, shaking up, filtering, precisely taking 5ml of subsequent filtrate, mixing the subsequent filtrate with 5ml of 10% sodium bisulfite solution, carrying out vortex oscillation for 5min, filtering, and taking the subsequent filtrate to obtain the test solution; (4) the measurement method comprises precisely sucking 5 μ l of each of the reference solution and the sample solution, injecting into liquid chromatograph, and measuring.
Chinese patent publication No. CN103739602A discloses a method for extracting matrine, which comprises the following steps: (1) preparing matrine molecular imprinting: dissolving 1-1.5mmol matrine and 6mmol methacrylic acid in 5-10ml chloroform, keeping at 0 deg.C for 30min, adding 15-30mmol ethylene glycol dimethacrylate, 15-30mmol N-isopropylacrylamide and 40-100mg azobisisobutyronitrile, mixing, introducing nitrogen, stirring to dissolve completely, introducing nitrogen for 10min, sealing, reacting at 65 deg.C until polymerization is complete, pulverizing, sieving with 200 mesh sieve, adding methanol: eluting matrine with acetic acid (9: 1), and loading to solid phase extraction column; (2) extracting the matrine solution: ultrasonic extracting radix Sophorae Tonkinensis with water as extraction medium, and filtering the extractive solution with 50nm ultrafiltration membrane; (3) extracting and separating matrine by a matrine molecular imprinting solid-phase extraction column: heating the membrane-passed extractive solution to 50 deg.C, loading on solid phase extraction column, eluting with 50 deg.C water to remove impurities, and eluting with 25 deg.C water to remove matrine.
Chinese patent publication No. CN107400133A discloses a method for preparing high-purity matrine from subprostrate sophora, which comprises the following steps: adding the subprostrate sophora powder into an ultrasonic extractor, adding a proper amount of reducing agent and ethanol, wherein the mass ratio of the reducing agent to the subprostrate sophora powder is 0.08-0.1: 1, the mass ratio of the ethanol to the subprostrate sophora powder is 10-20: 1, immersing for 30-60 min, performing ultrasonic reduction for 30-60 min at the temperature of 60-80 ℃ at the ultrasonic power of 300W, extracting for 4 times, combining extracting solutions, removing material residues, and concentrating the extracting solution; standing the concentrated solution overnight, filtering to remove starch impurities and part of tannin and protein, statically adsorbing the filtrate by macroporous resin H103, SP825, X-5 or D101, eluting with ethanol, concentrating the eluate to dryness, dissolving with water, acidifying the aqueous solution with concentrated hydrochloric acid or concentrated sulfuric acid to pH 3-4 or 2-3, salifying the alkaloid and dissolving in the aqueous solution of acid, dissolving part of low-polarity substances in water and separating out, filtering, and extracting the filtrate with petroleum ether to remove grease, wax and resin impurities. The petroleum ether layer was discarded and the acid aqueous layer was separated. Alkalizing the acid water layer with sodium hydroxide or potassium hydroxide until the pH value is 8-9, 9-10 or 10-10, so that the matrine is in a free state; extracting the alkaline water layer with petroleum ether under reflux for several times until no matrine is detected in the alkaline water layer. Concentrating the extract to 10 times of the volume of the medicinal materials, standing and crystallizing to obtain a refined matrine product with the purity of 97.9-98.5 percent; the reducing agent is sodium pyrosulfite, sodium sulfite, sodium bisulfite or potassium iodide; the concentration of the ethanol is 30-80% of the body tissue concentration.
Disclosure of Invention
The invention provides a method for preparing matrine from subprostrate sophora root, which comprises the following steps:
(1) crushing the vietnamese sophora root, adding an ethanol solution, carrying out high-temperature leaching at 80-90 ℃ for 1-3 h, and filtering to obtain a filtrate I and a filter residue I, wherein the weight ratio of the amount L of the added ethanol solution to the vietnamese sophora root is 1-3: 1;
(2) adding 5-15% of Aspergillus niger liquid into the filter residue I, fermenting for 3-8 days at room temperature, adding an ethanol solution, performing high-temperature leaching at 80-90 ℃ for 1-3 hours, and filtering to obtain a filtrate II, wherein the weight ratio of the amount L of the added ethanol solution to the subprostrate sophora ramosissima is 1-3: 1;
(3) combining the filtrate I and the filtrate II, concentrating the mixture at 50-70 ℃ under reduced pressure to 1/30-1/60 of the original volume, and extracting the mixture by using ethyl acetate with the volume being two to four times that of the original volume to obtain a water phase;
(4) adjusting the pH value of the water phase to 7.5-8, and extracting with butanol with a volume which is two to four times that of the water phase to obtain a butanol phase;
(5) concentrating the butanol phase at 50-70 ℃ under reduced pressure to 1/10-1/30 of the original volume, loading the butanol phase on an ADS-7 column, and eluting with ethanol solution four to six times the column volume to obtain eluent I, wherein the ethanol solution is 60-80% ethanol water solution;
(6) and concentrating the eluent I at 50-70 ℃ under reduced pressure to 1/10-1/30 of the original volume, crystallizing overnight at 4-8 ℃, and filtering to obtain the matrine product.
Adding 60-80% ethanol solution and 0.01-10 mmol/L hydrochloric acid into the ethanol solution in the steps (1) and (2).
The filtration is ceramic membrane filtration for 1 time, and the pore size of the membrane pore is 0.22 μm.
Adding Aspergillus niger liquid in the step (2), wherein the volume L/weight kg ratio of the subprostrate sophora tonkinensis is 5-10: 100;
technical effects
1. The matrine is efficiently released through aspergillus niger fermentation conversion, the yield is improved, and the production cost of the matrine is reduced.
2. The subprostrate sophora root resource is saved, and the environmental pollution emission is reduced.
Detailed Description
The present invention will be further described with reference to examples, but it is not limited to any one of these examples or the like.
Example 1
Taking 100kg of leaves of the subprostrate sophora, crushing, adding 300L of ethanol solution, carrying out high-temperature leaching at 85 ℃ for 3h, and filtering to obtain filtrate I and filter residue I; adding 15L of Aspergillus niger liquid into the filter residue I, fermenting for 6 days at a high temperature, adding 200L of ethanol solution, leaching for 3 hours at a high temperature of 85 ℃, and filtering to obtain a filtrate II; mixing the filtrate I and the filtrate II, concentrating at 60 deg.C under reduced pressure to 10L, and extracting with 30L ethyl acetate to obtain water phase; adjusting the pH value of the water phase to 8, and extracting with 20L butanol to obtain a butanol phase; concentrating butanol phase at 60 deg.C under reduced pressure to 1L, loading on ADS-7 column, eluting with 70% ethanol water solution to obtain 12L eluate I; concentrating the eluent I to 0.8L at 60 deg.C under reduced pressure, crystallizing at 4-8 deg.C overnight, and filtering to obtain 368g matrine product.
And (4) HPLC detection: the chromatographic column is Boston Green ODS PC18The mobile phase is acetonitrile-0.2% phosphoric acid aqueous solution, the flow rate is 0.8mL/min, the column temperature is 30 ℃, the detection wavelength is 220nm, and the sample injection amount is 10 mu L, wherein the volume L ratio of the acetonitrile to the 0.2% phosphoric acid aqueous solution is 0.04:0.76, 0.2% phosphoric acid aqueous solution containing 0.01% triethylamine is preferably selected from the 0.2% phosphoric acid aqueous solution, and the matrine content in a detection sample is 98.73%.
Example 2
Pulverizing 200kg leaves of Vietnamese sophora root, adding 600L ethanol solution, extracting at 85 deg.C for 3 hr, and filtering to obtain filtrate I and residue I; adding the filter residue I into 30L of an Aspergillus niger liquid chamber, carrying out warm fermentation for 6 days, adding 400L of ethanol solution, carrying out high-temperature leaching at 85 ℃ for 3 hours, and filtering to obtain a filtrate II; mixing the filtrate I and the filtrate II, concentrating at 60 deg.C under reduced pressure to 20L, and extracting with 60L ethyl acetate to obtain water phase; adjusting the pH value of the water phase to 8, and extracting with 40L butanol to obtain a butanol phase; concentrating butanol phase at 60 deg.C under reduced pressure to 1L, loading on ADS-7 column, and eluting with 70% ethanol water solution to obtain 20L eluate I; concentrating the eluent I at 60 ℃ under reduced pressure to 1.5L, crystallizing at 4-8 ℃ overnight, and filtering to obtain 735g of matrine product. The content of matrine in the sample is 98.82% by HPLC detection.
Example 3
Taking 400kg of leaves of the subprostrate sophora, crushing, adding 1000L of ethanol solution, carrying out high-temperature leaching at 85 ℃ for 3h, and filtering to obtain filtrate I and filter residue I; adding 50L of Aspergillus niger liquid into the filter residue I, fermenting for 6 days at a high temperature, adding 1000L of ethanol solution, leaching for 3 hours at a high temperature of 85 ℃, and filtering to obtain a filtrate II; mixing the filtrate I and the filtrate II, concentrating at 60 deg.C under reduced pressure to 40L, and extracting with 120L ethyl acetate to obtain water phase; adjusting the pH value of the water phase to 7.5, and extracting with 180L of butanol to obtain a butanol phase; concentrating butanol phase at 60 deg.C under reduced pressure to 4L, loading on ADS-7 column, and eluting with 70% ethanol water solution to obtain 40L eluate I; concentrating the eluent I to 2L at 60 deg.C under reduced pressure, crystallizing at 4-8 deg.C overnight, and filtering to obtain 1.68g matrine product. HPLC detection shows that the matrine content in the sample is 98.80%.
Claims (3)
1. A method for preparing matrine from subprostrate sophora root comprises the following steps:
(1) crushing the vietnamese sophora root, adding an ethanol solution, carrying out high-temperature leaching at 80-90 ℃ for 1-3 h, and filtering to obtain a filtrate I and a filter residue I, wherein the weight ratio of the amount L of the added ethanol solution to the vietnamese sophora root is 1-3: 1;
(2) adding 5-10% of an Aspergillus niger liquid chamber into the filter residue I, carrying out warm fermentation for 3-8 days, adding an ethanol solution, carrying out high-temperature leaching at 80-90 ℃ for 1-3 h, and filtering to obtain a filtrate II, wherein the weight ratio of the added ethanol solution L to the subprostrate sophora calyx is 1-3: 1;
(3) combining the filtrate I and the filtrate II, concentrating the mixture at 50-70 ℃ under reduced pressure to 1/30-1/60 of the original volume, and extracting the mixture by using ethyl acetate with the volume being two to four times that of the original volume to obtain a water phase;
(4) adjusting the pH value of the water phase to 7.5-8, and extracting with butanol with a volume which is two to four times that of the water phase to obtain a butanol phase;
(5) concentrating the butanol phase at 50-70 ℃ under reduced pressure to 1/10-1/30 of the original volume, loading the butanol phase on an ADS-7 column, and eluting with ethanol solution four to six times the column volume to obtain eluent I, wherein the ethanol solution is 60-80% ethanol water solution;
(6) and concentrating the eluent I at 50-70 ℃ under reduced pressure to 1/10-1/30 of the original volume, crystallizing overnight at 4-8 ℃, and filtering to obtain the matrine product.
2. The method according to claim 1, wherein the ethanol solution added in step (1) is 60-80% ethanol aqueous solution, and further contains 0.01-10 mmol/L hydrochloric acid.
3. The method according to claim 1, wherein the filtration is 1 filtration with a ceramic membrane having a pore size of 0.22 μm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811536654.0A CN111321179A (en) | 2018-12-15 | 2018-12-15 | Method for preparing matrine from subprostrate sophora |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811536654.0A CN111321179A (en) | 2018-12-15 | 2018-12-15 | Method for preparing matrine from subprostrate sophora |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111321179A true CN111321179A (en) | 2020-06-23 |
Family
ID=71165027
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811536654.0A Withdrawn CN111321179A (en) | 2018-12-15 | 2018-12-15 | Method for preparing matrine from subprostrate sophora |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111321179A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112174963A (en) * | 2020-10-13 | 2021-01-05 | 石家庄正道动物药业有限公司 | A method for processing matrine |
-
2018
- 2018-12-15 CN CN201811536654.0A patent/CN111321179A/en not_active Withdrawn
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112174963A (en) * | 2020-10-13 | 2021-01-05 | 石家庄正道动物药业有限公司 | A method for processing matrine |
| CN112174963B (en) * | 2020-10-13 | 2022-02-01 | 石家庄正道动物药业有限公司 | A method for processing matrine |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109568389B (en) | Preparation method of high-purity cannabinol extract | |
| CN101497594B (en) | Method for extracting, separating and purifying genistein from pigeon pea rhizome | |
| CN112870236B (en) | Flavone effective part of abelmoschus manihot and preparation method and application thereof | |
| CN103613679B (en) | A kind of gastrodia elata polysaccharide preparation method that is raw material with fresh rhizoma Gastrodiae | |
| CN101955479A (en) | Method for extracting orientin from bamboo leaf | |
| CN107501356A (en) | A kind of method that rutin is extracted from the sophora bud | |
| CN104940280A (en) | Method for extracting total flavones from radix puerariae employing enzyme preparation | |
| CN111321179A (en) | Method for preparing matrine from subprostrate sophora | |
| CN102477040A (en) | Method for preparing dendrobine | |
| CN108033970B (en) | Active extract of physalis alkekengi, extraction method and application | |
| CN101880303A (en) | Method for extracting bitter gourd saponin from bitter gourd | |
| CN105294793B (en) | The separation method of aurantiin in aizoon stonecrop | |
| CN104829474A (en) | Method for preparing glycine betaine chemical reference substances from boxthorn leaves | |
| Wu et al. | Preparation of paeoniflorin from the stems and leaves of Paeonia lactiflora Pall.‘Zhongjiang’through green efficient microwave-assisted extraction and subcritical water extraction | |
| CN101613278A (en) | A kind of from Folium Ipomoea the industrial technology of rapid extraction high-purity chlorogenic acid | |
| CN107213180B (en) | Separation and extraction method of notoginseng flavone | |
| CN108126000A (en) | Arasaponin extracts preparation method in fresh Radix Notoginseng | |
| CN112250655B (en) | Two novel cyclic diphenylheptanes compounds, preparation method and application thereof | |
| CN108997359A (en) | A method of chlorophyll is extracted from stevioside production waste residue | |
| CN105646638B (en) | The preparation method of pedunculoside | |
| KR20060098480A (en) | Formation of oil-soluble licorice extract including whitening bioactive materials | |
| CN111321181A (en) | Method for preparing N-formyl cytisine from subprostrate sophora | |
| CN111320628A (en) | Method for preparing cytisine from subprostrate sophora | |
| CN103665178B (en) | A kind of gastrodia elata polysaccharide extracting method | |
| CN101450962B (en) | Method for extracting oleanolic acid from Kandelia candel leaf |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20200623 |