CN110835297A - Method for extracting multiple active substances from waste liquid of ginkgo leaf extract production - Google Patents
Method for extracting multiple active substances from waste liquid of ginkgo leaf extract production Download PDFInfo
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- 239000007788 liquid Substances 0.000 title claims abstract description 41
- 239000009429 Ginkgo biloba extract Substances 0.000 title claims abstract description 23
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 20
- 238000000034 method Methods 0.000 title claims abstract description 19
- 239000002699 waste material Substances 0.000 title claims abstract description 18
- 239000013543 active substance Substances 0.000 title claims abstract description 7
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- 241000218628 Ginkgo Species 0.000 claims abstract description 52
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 43
- JXOHGGNKMLTUBP-HSUXUTPPSA-N shikimic acid Chemical compound O[C@@H]1CC(C(O)=O)=C[C@@H](O)[C@H]1O JXOHGGNKMLTUBP-HSUXUTPPSA-N 0.000 claims abstract description 33
- JXOHGGNKMLTUBP-JKUQZMGJSA-N shikimic acid Natural products O[C@@H]1CC(C(O)=O)=C[C@H](O)[C@@H]1O JXOHGGNKMLTUBP-JKUQZMGJSA-N 0.000 claims abstract description 33
- 239000013078 crystal Substances 0.000 claims abstract description 26
- 238000001556 precipitation Methods 0.000 claims abstract description 17
- 150000004676 glycans Chemical class 0.000 claims abstract description 16
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 16
- 239000005017 polysaccharide Substances 0.000 claims abstract description 16
- 239000000284 extract Substances 0.000 claims abstract description 11
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 10
- 238000002425 crystallisation Methods 0.000 claims abstract description 10
- 230000008025 crystallization Effects 0.000 claims abstract description 10
- 150000001450 anions Chemical class 0.000 claims abstract description 8
- 239000000463 material Substances 0.000 claims abstract description 5
- 238000010828 elution Methods 0.000 claims abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 25
- 239000000243 solution Substances 0.000 claims description 20
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 16
- 239000002244 precipitate Substances 0.000 claims description 16
- 238000005406 washing Methods 0.000 claims description 13
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 12
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 12
- 235000019750 Crude protein Nutrition 0.000 claims description 10
- 239000003480 eluent Substances 0.000 claims description 10
- 239000012535 impurity Substances 0.000 claims description 10
- 238000011068 loading method Methods 0.000 claims description 10
- 239000011347 resin Substances 0.000 claims description 9
- 229920005989 resin Polymers 0.000 claims description 9
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 8
- 239000007864 aqueous solution Substances 0.000 claims description 8
- 238000004440 column chromatography Methods 0.000 claims description 8
- 235000019253 formic acid Nutrition 0.000 claims description 8
- 229940068052 ginkgo biloba extract Drugs 0.000 claims description 7
- 235000020686 ginkgo biloba extract Nutrition 0.000 claims description 7
- 239000003153 chemical reaction reagent Substances 0.000 claims description 6
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- 238000002156 mixing Methods 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 5
- 238000001704 evaporation Methods 0.000 claims description 4
- 238000001179 sorption measurement Methods 0.000 claims description 4
- 239000002994 raw material Substances 0.000 claims description 3
- 238000001953 recrystallisation Methods 0.000 claims description 2
- 238000000108 ultra-filtration Methods 0.000 claims 1
- 244000194101 Ginkgo biloba Species 0.000 abstract description 5
- 101710138460 Leaf protein Proteins 0.000 abstract description 4
- 108090000623 proteins and genes Proteins 0.000 abstract description 3
- 102000004169 proteins and genes Human genes 0.000 abstract description 3
- 238000004064 recycling Methods 0.000 abstract description 2
- 238000004065 wastewater treatment Methods 0.000 abstract description 2
- 239000000047 product Substances 0.000 description 9
- 238000000605 extraction Methods 0.000 description 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
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- 238000001035 drying Methods 0.000 description 5
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- 238000000643 oven drying Methods 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
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- 238000010992 reflux Methods 0.000 description 2
- 210000000582 semen Anatomy 0.000 description 2
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- 238000005303 weighing Methods 0.000 description 2
- HVAUUPRFYPCOCA-AREMUKBSSA-N 2-O-acetyl-1-O-hexadecyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCCOC[C@@H](OC(C)=O)COP([O-])(=O)OCC[N+](C)(C)C HVAUUPRFYPCOCA-AREMUKBSSA-N 0.000 description 1
- SQGLUEWZRKIEGS-UHFFFAOYSA-N Ginkgetin Natural products C1=CC(OC)=CC=C1C1=CC(=O)C2=C(O)C=C(OC)C(C=3C(=CC=C(C=3)C=3OC4=CC(O)=CC(O)=C4C(=O)C=3)O)=C2O1 SQGLUEWZRKIEGS-UHFFFAOYSA-N 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 229930045534 Me ester-Cyclohexaneundecanoic acid Natural products 0.000 description 1
- 206010052779 Transplant rejections Diseases 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
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- 238000006243 chemical reaction Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- AIFCFBUSLAEIBR-UHFFFAOYSA-N ginkgetin Chemical compound C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C(C=1)=CC=C(OC)C=1C1=C(O)C=C(O)C(C(C=2)=O)=C1OC=2C1=CC=C(O)C=C1 AIFCFBUSLAEIBR-UHFFFAOYSA-N 0.000 description 1
- 229930184727 ginkgolide Natural products 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
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- 230000001681 protective effect Effects 0.000 description 1
- 239000003087 receptor blocking agent Substances 0.000 description 1
- 210000004994 reproductive system Anatomy 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
- C07C51/42—Separation; Purification; Stabilisation; Use of additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/006—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from vegetable materials
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/16—Ginkgophyta, e.g. Ginkgoaceae (Ginkgo family)
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- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
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Abstract
The invention discloses a method for extracting various active substances from waste liquid in the production of ginkgo leaf extract, which comprises the following steps: (1) alcohol precipitation of waste liquid from ginkgo leaf extract production; (2) processing alcohol precipitation; (3) processing an alcohol precipitation solution; (4) recrystallizing shikimic acid crude crystal. Compared with the prior art, the invention can effectively and continuously extract 4 products of ginkgo leaf extract, ginkgo leaf polysaccharide, ginkgo leaf protein and shikimic acid in ginkgo leaves, and improve the comprehensive utilization degree of ginkgo leaf medicinal materials; comprehensively recycling the ginkgo biloba polysaccharides, the ginkgo biloba proteins and the shikimic acid, thereby reducing the cost of wastewater treatment in the production process; when the column is washed, double columns of anions and activated carbon columns are connected in series, and the decoloring treatment is carried out while the elution is carried out, so that the secondary decoloring procedure during the crystallization is reduced, and the production efficiency is improved. Is suitable for comprehensive processing and utilization of ginkgo leaves.
Description
Technical Field
The invention relates to multi-stage treatment of wastewater, in particular to a method for extracting ginkgo leaves and comprehensively utilizing waste liquid thereof to obtain various active substances.
Technical Field
As is well known, Ginkgo biloba is a leaf of Ginkgo biloba (Ginkgo biloba L.) belonging to the genus Ginkgo of the family Ginkgo. The ginkgo biloba extract has unique physiological and pharmacological effects and therapeutic values, and particularly, the ginkgolide is a PAF (platelet activating factor) receptor blocker with high specificity, so that the ginkgo biloba extract is well appreciated by people. The literature considers that the main pharmacological actions of the ginkgo biloba extract are as follows: has effects in treating central nervous system, resisting shock, and resisting allergy. The protective action for organ transplant rejection reaction, besides the above-mentioned action, has the inhibiting action for cell immunological reaction, eye pathological change and circulation change due to endotoxin, and also has the action for reproductive system. The ginkgo biloba extract is developed in China since the last 70 years, but the research on the comprehensive utilization of the ginkgo biloba extract waste liquid is not much.
At present, in the Chinese patent database, there are 2 patent application pieces relating to ginkgo leaf extraction wastewater: CN106495411A No. ICBB technology and device for treating wastewater in the extraction process of folium Ginkgo extract, and CN110002616A No. device and method for treating and recycling wastewater from folium Ginkgo extraction of ginkgetin, which are both treatment technologies for folium Ginkgo extraction wastewater. In addition, there are 3 patent applications relating to the preparation of shikimic acid from waste liquid of ginkgo leaf extraction: CN109593034A A method for preparing shikimic acid from waste liquid of ginkgo leaf extraction, CN108191639A A method for preparing shikimic acid extract from waste liquid of ginkgo leaf extract chromatography, and CN109369373A A method for preparing shikimic acid extract from waste liquid of ginkgo leaf extract chromatography are all technical schemes for preparing shikimic acid. At present, no patent application for comprehensive utilization of ginkgo leaf extraction waste liquid exists.
Disclosure of Invention
The invention aims to provide a method for extracting various active substances from ginkgo leaf extract production waste liquid, which not only changes the ginkgo leaf extract production waste liquid into valuable and reduces the pollution to the environment, but also can obtain ginkgo leaf polysaccharide, ginkgo leaf protein and shikimic acid products and realize comprehensive utilization.
The method for extracting various active substances from the production waste liquid of the ginkgo leaf extract provided by the inventor comprises the following steps:
(1) alcohol precipitation of waste liquid from production of ginkgo leaf extract
Macroporous resin sample loading residual liquid and water washing liquid generated in the production process of the ginkgo biloba extract prepared according to Chinese pharmacopoeia are concentrated to the relative density of the material liquid of about 1.1, thus obtaining the raw material liquid. Carrying out alcohol precipitation by adopting ethanol to obtain a white precipitate and an alcohol precipitation solution for later use;
(2) processing alcohol precipitation
Dissolving the white precipitate in water, ultrafiltering the obtained solution with molecular weight of 5000 or 1 ten thousand, vacuum concentrating the concentrated solution to 200L, adding Sevage reagent, mixing, oscillating, centrifuging to obtain folium Ginkgo crude protein precipitate, and evaporating the centrifugate to dryness to obtain folium Ginkgo crude polysaccharide;
(3) processing of alcohol precipitation liquid
Concentrating the alcohol precipitation solution until no alcohol smell exists, then carrying out column chromatography on the treated anion resin, slowly loading the extract above an adsorption column, adsorbing for a period of time, washing with distilled water to remove impurities, connecting an activated carbon column in series after removing the impurities, eluting the column in series with formic acid aqueous solution, collecting eluent, concentrating the eluent to a small volume, placing the concentrated eluent in an environment at 0-25 ℃ for crystallization, and obtaining shikimic acid crude crystals with the purity of more than 80%;
(4) recrystallization of crude crystals of shikimic acid
Adding methanol or water into the shikimic acid crude crystal for dissolving, and then placing the solution in the temperature environment the same as the temperature environment of the crude crystal for secondary crystallization to obtain the shikimic acid crystal with the purity of more than 98 percent.
In the step (1), the mass fraction of the ethanol is 90-95%.
In the step (2), the amount of the added water is controlled to enable the ratio of the materials to the liquid to be 1: 10-1: 20; the vacuum degree of the vacuum concentration control is 0.06-0.09 MPa; the Sevage reagent is a mixture of chloroform and n-butanol, wherein the volume ratio of the chloroform to the n-butanol is 4: 1.
In the step (3), the sample loading speed is 1-2 times of column volume/h; the amount of the distilled water for washing and removing impurities is 2-4 times of the column volume; the mass fraction of the formic acid aqueous solution of the elution string column is 3% -8%, and the using amount of the formic acid aqueous solution is 3-5 times of the column volume.
In the step (4), the amount of the added methanol or water is 1/5-1/10 of the amount of the shikimic acid crude crystals; the same temperature as the crude crystal is 0 ℃ to 25 ℃.
The inventor indicates that the ginkgo leaf crude protein precipitate and the ginkgo leaf crude polysaccharide prepared in the step (2) can be further processed and purified by adopting a conventional process according to the market demand to prepare a pure product.
The innovation points of the invention are as follows:
(1) the invention can effectively and continuously extract 4 products of ginkgo leaf extract, ginkgo leaf polysaccharide, ginkgo leaf protein and shikimic acid in ginkgo leaves, improves the comprehensive utilization rate of ginkgo leaf medicinal materials, lowers the comprehensive production cost of the products and effectively improves the market competitiveness of production plants.
(2) In the production process of the ginkgo leaf extract, ginkgo leaf polysaccharide, ginkgo leaf protein and shikimic acid are comprehensively recycled from waste wastewater, so that the wastewater treatment pressure in the production process is reduced.
(3) When the column is washed, double columns of anions and activated carbon columns are connected in series, so that the decolorization treatment is carried out while enrichment and elution are carried out, the secondary decolorization process during crystallization is reduced, and the production efficiency is improved.
The invention is suitable for comprehensive processing and utilization of ginkgo leaf extract produced by ginkgo leaves.
Drawings
FIG. 1 is a crystal pattern of shikimic acid,
FIG. 2 is a raw material liquid spectrum,
FIG. 3 is a map of shikimic acid in ginkgo leaves.
Detailed Description
Example 1
Weighing folium Ginkgo 1t, adding 70% ethanol, and extracting under reflux for 2 times; mixing the extractive solutions, concentrating until no alcohol smell exists, adding 3 times of water, stirring, and precipitating; filtering to obtain clear liquid, performing DM130 column chromatography on the filtered clear liquid, washing with 2 times of column volume of water, and collecting effluent liquid and washing liquid of the column for later use; eluting with 70% ethanol water solution, deacidifying the eluate with deacidifying resin, concentrating, and oven drying to obtain folium Ginkgo extract under the item of 2015 edition of Chinese pharmacopoeia;
collecting column chromatography column effluent and water washing solution 3 tons, concentrating to relative density of 1.1, adding 95% ethanol solution 9 times, precipitating with ethanol, filtering, collecting white flocculent precipitate, separating and purifying folium Ginkgo polysaccharide and semen Ginkgo protein, and separating shikimic acid from filtrate.
Dissolving the white precipitate in water at a ratio of 1: 20, ultrafiltering the solution with 1 ten thousand molecular weight, concentrating the concentrated solution to about 200L under a vacuum degree of about 0.08MPa, adding Sevage reagent (the volume ratio of chloroform to n-butanol is 4: 1), mixing, oscillating, centrifuging to obtain crude protein precipitate of folium Ginkgo, evaporating the centrifugate to obtain crude polysaccharide of folium Ginkgo, drying the crude protein precipitate of folium Ginkgo with two products to obtain about 80kg, and drying the crude protein precipitate of folium Ginkgo with the two products to obtain about 60kg crude polysaccharide of folium Ginkgo. The ginkgo leaf crude protein and the ginkgo leaf crude polysaccharide are directly applied or further processed according to the requirements of users.
Concentrating the alcohol precipitation filtrate until no alcohol smell exists, performing column chromatography on anion resin which is treated by about 500L, slowly loading the extract above an adsorption column (the loading speed is 1 time of column volume/h), washing with distilled water with the volume of 3 times to remove impurities, connecting the anion column in series with a 300L active carbon column after the impurities are removed, eluting with formic acid aqueous solution with the concentration of 4% with the volume of 5 times, collecting eluent, concentrating the eluent to 80L, placing in a room temperature environment for crystallization to obtain shikimic acid crude crystals, and pumping out the crude crystals by vacuum filtration. Adding the crude crystals into water according to the feed-liquid ratio of 1: 5 for dissolving, placing the mixture in the same temperature environment for secondary crystallization to obtain white granular shikimic acid crystals with the purity of more than 98%, and performing suction filtration and drying to obtain 98% white shikimic acid crystals.
Example 2
Weighing 1 ton of ginkgo leaves, adding 70% ethanol for reflux extraction, and extracting for 2 times; mixing the extractive solutions, concentrating until no alcohol smell exists, adding 4 times of water, stirring, and precipitating; filtering to obtain clear liquid, performing D101 column chromatography on the filtered clear liquid, washing with 3 times of column volume of water, and collecting effluent liquid and washing liquid of the column for later use; eluting with 68% ethanol water solution, deacidifying the eluate with deacidifying resin, concentrating, and oven drying to obtain folium Ginkgo extract under the item of 2015 edition of Chinese pharmacopoeia;
collecting eluate and water eluate 4t after resin column chromatography, concentrating to relative density of 1.1, adding 95% ethanol 10 times of the feed liquid, precipitating with ethanol, filtering, collecting white flocculent precipitate, separating and purifying folium Ginkgo polysaccharide and semen Ginkgo protein, and separating shikimic acid from the filtrate.
Dissolving the white precipitate in water at ratio of 1: 15, ultrafiltering the solution with 1 ten thousand molecular weight, concentrating the concentrated solution at vacuum degree of about 0.08MPa to about 150L, adding Sevage reagent (mass ratio of chloroform to n-butanol is 4: 1), mixing, oscillating, and centrifuging to obtain crude protein precipitate of folium Ginkgo; evaporating the centrifugate to dryness to obtain folium Ginkgo crude polysaccharide, drying folium Ginkgo crude protein precipitate of the two products to about 75kg, and folium Ginkgo crude polysaccharide to about 55 kg. The ginkgo leaf crude protein and the ginkgo leaf crude polysaccharide are directly applied or further processed according to the requirements of users.
Concentrating the alcohol precipitation filtrate until no alcohol smell exists, performing column chromatography on anion resin which is treated by about 600L, slowly loading the extract above an adsorption column (the loading speed is 1 column volume/h), washing with distilled water with the volume of 3 times to remove impurities, connecting the anion column in series with a 300L active carbon column after the impurities are removed, eluting with 4 times volume of formic acid aqueous solution with the concentration of 4%, collecting eluent, concentrating the eluent to 80L, placing in a room temperature environment for crystallization to obtain shikimic acid crude crystals, and pumping out the crude crystals by vacuum filtration. Adding 50% methanol water into the crude crystals according to the feed-liquid ratio of 1: 5 for dissolving, placing the mixture in the same temperature environment for secondary crystallization to obtain white granular shikimic acid crystals with the purity of more than 98%, and performing suction filtration and drying to obtain 98% white shikimic acid crystals.
TABLE 1 content of shikimic acid in samples at different stages of the process
Table 2 table of product quality for different production batches:
shikimic acid product quality analysis report
Quality analysis report of shikimic acid
Claims (5)
1. A method for extracting a plurality of active substances from ginkgo leaf extract production waste liquid is characterized by comprising the following steps:
(1) alcohol precipitation of waste liquid from production of ginkgo leaf extract
Combining macroporous resin sample loading residual liquid and water washing liquid generated in the production process of the ginkgo biloba extract prepared according to Chinese pharmacopoeia, concentrating the mixture until the relative density of the material liquid is 1.1 to obtain a raw material liquid;
carrying out alcohol precipitation by adopting high-concentration ethanol to obtain a white precipitate and an alcohol precipitation solution for later use;
(2) processing alcohol precipitation
Dissolving the white precipitate in water, performing ultrafiltration on the obtained solution with the molecular weight of 1 ten thousand, further performing vacuum concentration on the concentrated solution to obtain about 150-200L, adding a Sevage reagent, mixing, oscillating, centrifuging to obtain a ginkgo leaf crude protein precipitate, and evaporating the centrifugate to dryness to obtain a ginkgo leaf crude polysaccharide;
(3) processing of alcohol precipitation liquid
Concentrating the alcohol precipitation solution until no alcohol smell exists, then carrying out column chromatography on the treated anion resin, slowly loading the extract above an adsorption column, adsorbing for a period of time, washing with distilled water to remove impurities, connecting an activated carbon column in series after removing the impurities, eluting the column in series with formic acid aqueous solution, collecting eluent, concentrating the eluent to a small volume, and placing the concentrated eluent in an environment at 0-25 ℃ for crystallization to obtain shikimic acid crude crystals;
(4) recrystallization of crude crystals of shikimic acid
Adding a proper amount of water into the shikimic acid crude crystal for dissolving, and then placing the solution in the temperature environment same as the crude crystal for secondary crystallization to obtain shikimic acid crystal with the purity of more than 98 percent.
2. The method according to claim 1, wherein the ethanol is present in an amount of 90 to 95% by weight in step (1).
3. The method according to claim 1, wherein in the step (2), the water is added in an amount such that the ratio of the feed to the liquid is 1: 10 to 1: 20; the vacuum degree of the vacuum concentration control is 0.06-0.09 MPa; the Sevage reagent is a mixture of chloroform and n-butanol, wherein the volume ratio of the chloroform to the n-butanol is 4: 1.
4. The method according to claim 1, wherein in the step (3), the sample loading speed is 1-2 times of column volume/h; the amount of the distilled water for washing and removing impurities is 2-4 times of the column volume; the mass fraction of the formic acid aqueous solution of the elution string column is 3% -8%, and the using amount of the formic acid aqueous solution is 3-5 times of the column volume.
5. The method according to claim 1, wherein in the step (4), the amount of the added water is 1/5-1/10% of the crude crystals; the same temperature as the crude crystal is 0 ℃ to 25 ℃.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111233658A (en) * | 2020-02-27 | 2020-06-05 | 陕西嘉禾生物科技股份有限公司 | Method for extracting shikimic acid and quinic acid from folium ginkgo |
| CN111978422A (en) * | 2020-08-25 | 2020-11-24 | 湖南华诚生物资源股份有限公司 | Method for separating multiple active ingredients from waste liquid after pigment extraction from roselle |
| CN112390715A (en) * | 2020-11-13 | 2021-02-23 | 晨光生物科技集团股份有限公司 | Method for extracting shikimic acid and quinic acid from ginkgo leaf extract adsorbed lower column water |
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| CN113698289A (en) * | 2020-05-21 | 2021-11-26 | 晨光生物科技集团股份有限公司 | Method for preparing shikimic acid from ginkgo leaf extraction waste liquid |
| CN114057572A (en) * | 2020-08-03 | 2022-02-18 | 江苏得乐康生物科技有限公司 | Method for simultaneously extracting shikimic acid and quinic acid from ginkgo leaves |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104031160A (en) * | 2014-06-26 | 2014-09-10 | 河南中烟工业有限责任公司 | Gingko polysaccharide, extraction and purification method and application of gingko polysaccharide as tobacco humectant |
| CN107353201A (en) * | 2016-05-09 | 2017-11-17 | 云南希康生物科技有限公司 | A kind of natural shikimic acid extract of high content and preparation method thereof |
| CN109593034A (en) * | 2018-11-22 | 2019-04-09 | 桂林莱茵生物科技股份有限公司 | A method of it is extracted in waste liquid from ginkgo leaf and prepares shikimic acid |
-
2019
- 2019-10-29 CN CN201911033149.9A patent/CN110835297A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104031160A (en) * | 2014-06-26 | 2014-09-10 | 河南中烟工业有限责任公司 | Gingko polysaccharide, extraction and purification method and application of gingko polysaccharide as tobacco humectant |
| CN107353201A (en) * | 2016-05-09 | 2017-11-17 | 云南希康生物科技有限公司 | A kind of natural shikimic acid extract of high content and preparation method thereof |
| CN109593034A (en) * | 2018-11-22 | 2019-04-09 | 桂林莱茵生物科技股份有限公司 | A method of it is extracted in waste liquid from ginkgo leaf and prepares shikimic acid |
Non-Patent Citations (1)
| Title |
|---|
| 刘东彦 等: "雪松松针中莽草酸和多糖的提取工艺研究", 《中药材》, vol. 40, no. 8, pages 1914 - 1916 * |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111233658B (en) * | 2020-02-27 | 2022-06-07 | 陕西嘉禾生物科技股份有限公司 | Method for extracting shikimic acid and quinic acid from folium ginkgo |
| CN111233658A (en) * | 2020-02-27 | 2020-06-05 | 陕西嘉禾生物科技股份有限公司 | Method for extracting shikimic acid and quinic acid from folium ginkgo |
| CN113511967A (en) * | 2020-04-10 | 2021-10-19 | 江苏得乐康生物科技有限公司 | Method for extracting quinic acid from ginkgo leaf extract chromatographic wastewater |
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