CN111676264A - Method for extracting caffeic acid from rosemary - Google Patents
Method for extracting caffeic acid from rosemary Download PDFInfo
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- CN111676264A CN111676264A CN202010596385.8A CN202010596385A CN111676264A CN 111676264 A CN111676264 A CN 111676264A CN 202010596385 A CN202010596385 A CN 202010596385A CN 111676264 A CN111676264 A CN 111676264A
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- 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
- C12P39/00—Processes involving microorganisms of different genera in the same process, simultaneously
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- 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
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/42—Hydroxy-carboxylic acids
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Abstract
The invention discloses a method for extracting caffeic acid from rosemary, which comprises the following steps: (1) crushing rosemary, extracting with ethanol at 80-83 ℃ to obtain an extracting solution, and carrying out solid-liquid separation on the extracting solution to obtain a clear extracting solution; (2) concentrating the clear extracting solution to Brix 40-60%, cooling, and adding a carbon source; (3) inoculating the mixed bacteria liquid consisting of lactic acid bacteria and saccharomycetes into the step (2), adjusting the pH value, and fermenting to obtain liquid; (4) adding water into the liquid until the Brix 11-12% is reached, adjusting the pH value to 4-6, and carrying out solid-liquid separation to obtain a clarified liquid; (5) adding the clarified liquid into macroporous adsorption resin, washing the macroporous adsorption resin, and concentrating under reduced pressure until Brix 30-35%; (6) decolorizing and filtering the concentrated solution, concentrating again, and drying to obtain caffeic acid extract. The method has high effective component content, environment friendliness, high extraction efficiency, low cost, and suitability for industrial production.
Description
Technical Field
The invention relates to a method for extracting caffeic acid from rosemary, and belongs to the technical field of biological medicines.
Background
Rosmarinus officinalis is a plant of Rosmarinus of Labiatae. The rosemary is a plant with abundant use value, is originally produced in mediterranean regions, is widely cultivated in countries in the world at present, and is used for enhancing memory, treating language disorder, conditioning menstruation, growing hair, promoting digestion and protecting liver in European countries more than 2000 years ago. Rosemary, recorded in Bencao Shi Yi, is pungent, warm and non-toxic. Mainly for foul breath. At present, China has large-area planting in Yunnan, Guizhou, Guangxi, Hainan, Xinjiang and other places.
For the plant research, a large number of reports and patents are provided abroad, and the components of the plant mainly comprise diterpene phenols, flavonoids, triterpenes, essential oil and the like, and the plant is widely applied to the aspects of medicines, cosmetics, foods and the like. Rosemary is also widely used as food additive, and its antibacterial and antioxidant effects can improve food quality and prolong fresh-keeping time. In recent years, the interest of rosemary has been increasing, and many reports have been made about the chemical components and pharmacological actions of rosemary. Rosemary contains rosmarinic acid which is absorbed and metabolized in vivo in a conjugate or methylated form, and most of the absorbed rosmarinic acid is degraded into a conjugate of caffeic acid, ferulic acid and cinnamic acid, or methylated form, and then gradually secreted into urine. Has various pharmacological actions such as antibacterial action, anti-inflammatory action, antivirus action, antioxidation action, anti-apoptosis action, free radical removal action, immunosuppression action and the like, particularly has the protection action on myocardial cell injury, nerve cell apoptosis resistance, T cell apoptosis induction and anticancer action, has strong amyloid generation resistance on Alzheimer beta-amyloid fibrils, and the like, so that the application prospect and market value of the novel anti-tumor medicine are very wide.
Caffeic acid is also called hydrolytic caffeotannic acid, 3, 4-dihydroxy cinnamic acid, 3- (3, 4-dihydroxy phenyl) -2-acrylic acid, and is cinnamic acid compound; it is yellow crystal, slightly soluble in cold water, and easily soluble in hot water and cold ethanol. Is commonly found in common plant herbs such as radix Angelicae sinensis and rhizoma Ligustici Chuanxiong, and is commonly found in flowers, leaves or fruits of plants of Compositae, Rosaceae, Sapindaceae, Ranunculaceae, Polypodiaceae, Rutaceae, Labiatae and Eucommiaceae. Has antibacterial, antiviral, anti-snake venom, anti-diarrhea, anti-fertility, and anti-tumor pharmacological activities, and also has blood cooling and hemostatic effects, and can be used for treating hypertension, coronary heart disease, arrhythmia, etc. Caffeic acid, a representative compound of hydroxycinnamic acids, has a strong antioxidant activity.
At present, no report of extracting and preparing caffeic acid from rosemary is found. Mainly by extracting caffeic acid from plants rich in caffeic acid, which is usually not high in content, the direct extraction of caffeic acid from rosemary is more rarely harvested, so that a new way for obtaining caffeic acid is needed.
The research finds that rosemary contains more rosmarinic acid and a small amount of caffeic acid, the invention provides a preparation method of caffeic acid, rosmarinic acid is extracted and then is converted into caffeic acid through fermentation, so that the yield of caffeic acid obtained from rosemary is greatly improved, and the efficient utilization of rosemary resources is realized.
Disclosure of Invention
The invention aims to provide an efficient and economic method for extracting caffeic acid from rosemary, and simultaneously improve the content of the caffeic acid in the extract.
A method for extracting caffeic acid from rosemary is characterized by comprising the following steps:
s1, crushing rosemary, extracting with ethanol at 80-83 ℃, and carrying out solid-liquid separation on the extracting solution to obtain a clear extracting solution;
s2, concentrating the clear extracting solution obtained in the step S1 to Brix 40-60% to obtain a concentrated solution, and adding a carbon source;
s3, inoculating zymophyte into the concentrated solution added with the carbon source in the step S2, adjusting the pH value to 4-6, and fermenting to obtain caffeic acid liquid;
s4, adding water into the caffeic acid liquid obtained in the step S3 to Brix 11-12%, adjusting the pH value to 4-6, and performing solid-liquid separation to obtain clear supernatant;
s5, adding the supernatant obtained in the step S4 into macroporous adsorption resin, and washing the macroporous adsorption resin with water to remove unadsorbed impurities; washing the macroporous adsorption resin with desorption solution with the pH value of 4-6, collecting the washed desorption solution, and concentrating under reduced pressure to Brix 30-35%;
s6, decolorizing and filtering the concentrated solution, concentrating the filtrate again, and drying to obtain the caffeic acid extract.
Preferably, in the step S1, rosemary is crushed to a diameter of 0.5cm or less.
Preferably, the ethanol concentration in the step S1 is 50% to 60%.
The ethanol concentration is 50% -60% because the solubility of the rosemary caffeic acid is relatively proper at the concentration, so that the alcohol-soluble impurities brought into the rosemary caffeic acid as little as possible can be avoided, the burden of the post-treatment process is increased, and the production efficiency is improved.
Preferably, the extraction of step (1) is a percolation extraction.
Preferably, the carbon source is glucose.
Preferably, the solid-liquid separation is filtration or centrifugation.
Preferably, the fermentation bacteria comprise, by mass, 10-90% of lactic acid bacteria and 10-90% of yeast.
Preferably, the fermentation bacteria comprise 60-80% of lactic acid bacteria and 20-40% of yeast by mass percentage.
Preferably, the fermentation bacteria comprise 70% of lactic acid bacteria and 30% of yeast in percentage by mass.
Caffeine is a xanthine alkaloid compound, caffeine is prepared through biotransformation, lactic acid bacteria and saccharomycetes are the most effective biological bacteria for plant biological fermentation, the lactic acid bacteria and the saccharomycetes are probiotic bacteria, a fermentation process of generating lactic acid is theoretically realized through fermentation of a large number of lactic acid bacteria and anaerobic glycolysis, and ethanol fermentation is two main fermentation forms in an organism, so that smooth fermentation can be promoted. And a small amount of yeast is added for fermentation, so that the sugar is decomposed into carbon dioxide and water under the aerobic condition, the growth of the yeast is fast, and the sugar is decomposed into alcohol and carbon dioxide under the anoxic condition, so that the fermentation process is expanded, and the bioconversion process of caffeine is facilitated. Therefore, it is preferable that the ratio of lactic acid bacteria is larger.
Preferably, the fermentation temperature is 35-37 ℃ and the period is 7-10 days.
Preferably, the pH adjusted solution is dilute hydrochloric acid.
Preferably, the macroporous adsorption resin is NKA-9 macroporous resin, D1O1 macroporous resin, D301 macroporous basic resin, D201 macroporous basic resin and 001 × 7Na 732.
Preferably, the macroporous adsorbent resin is a medium-polarity macroporous adsorbent resin.
Preferably, the macroporous adsorption resin is NKA-9 macroporous resin or D1O1 macroporous resin.
More preferably, the macroporous adsorption resin is NKA-9 macroporous adsorption resin.
The NKA-9 macroporous adsorption resin is an adsorbent with divinylbenzene as a framework structure, a benzene ring connected to a main chain is a plane with uniformly distributed electrons, and the NKA-9 macroporous adsorption resin has strong adsorption capacity for molecules with similar properties and various cyclic aromatic compounds and is increased along with the enhancement of lipophilicity of adsorbed molecules. Is widely used for separating ginkgo flavone, polyphenol, caffeic acid and the like. In recent years, it has been shown to have a unique effect on the separation and purification of natural products, particularly water-soluble compounds.
Preferably, the desorption solution is an ethanol solution with the volume percentage concentration of 50-60%.
Preferably, the active carbon filter cloth for decolorization and filtration has a 325-mesh screen.
Preferably, the drying is vacuum drying or spray drying until the water content of the caffeic acid extract is not more than 5%.
At present, the similar substances are usually extracted by alcohol, and are precipitated at low temperature for a long time after being concentrated to remove impurities, and the main defects are high temperature requirement (below 4 ℃) and incomplete impurity removal, so that the production cost is high, the content of active ingredients is low, and the industrialization is difficult. The caffeic acid prepared by the traditional chemical synthesis method has the defects of long reaction time, unsatisfactory purity, high residue of harmful substances in the product, unsatisfactory caffeic acid product and the like. The method extracts the rosmarinic acid and the caffeic acid by 60-65% ethanol, and converts the rosmarinic acid into the caffeic acid by fermentation after concentration, thereby greatly improving the yield of obtaining the caffeic acid from the rosemary and realizing the efficient utilization of rosemary resources. Before resin column separation, the invention also adds water into the obtained liquid to Brix 10-12%, and adjusts the pH value to 4-6 with acid, under the condition, the impurities in the solution are rapidly precipitated under the acidic condition, and do not need to be stood for a long time at low temperature, and are separated immediately, thereby greatly shortening the production period. Separating with macroporous adsorbent resin column to obtain the product with high effective component content. The concentrated solution is decolorized and filtered by 325-mesh active carbon filter cloth, and partial colored substances and water-insoluble impurities can be removed. In the conventional extraction method, the content of a target product in the extract is generally 10-20%, but the content of caffeic acid in the extract can reach 50-60% by using the method provided by the invention. The product prepared by the invention is mainly used as a raw material of medicines.
Compared with the prior art, the invention has the following beneficial effects:
1. the method has the advantages of good extraction continuity of the caffeic acid, simple process and suitability for industrial popularization.
2. The method comprises the step of fermenting rosemarinic acid and mixed bacteria consisting of lactic acid bacteria and yeast at the pH value of 4-6 and the Brix 40-60% to obtain the caffeic acid, wherein the fermentation process contains 6-P-beta-glucosidase, Gallate decarboxylase and P-coumaric acid carboxylase which participate in phenolic compound metabolism to convert the rosemarinic acid into the caffeic acid, so that the content of the caffeic acid is improved.
3. The invention adopts a leakage method to extract caffeic acid, so that the effective components in the rosemary can be quickly and fully dissolved out in a short time, the reaction conditions are mild, the structure protection of active substances is facilitated, new organic impurities cannot be introduced, the safety of final products is ensured, and the environmental pollution is reduced.
4. The method has the advantages of reasonable process, high content of effective components, high extraction efficiency, simple operation, and suitability for industrial production.
5. The extract prepared by the invention has stable purity, and the caffeic acid content is 50-60%.
6. The solvent of the invention can be recycled, and is economic and environment-friendly.
Drawings
FIG. 1 is a graph of the effect of caffeic acid on ethanol extraction of various concentrations selected in example 1;
FIG. 2 is a graphical analysis of the conditions for influencing fermentation broths of different ratios with caffeic acid rosemary in example 1;
wherein the ratio of lactic acid bacteria: the yeast mixture ratio is A1:9, B3:7 and C5: 5. d7: 3. e9: 1;
FIG. 3 is a graph of the selective elution of caffeic acid from rosemary in example 2 under different conditions on macroporous adsorbent resins;
wherein A is NKA-9 macroporous resin, B is D1O1 macroporous resin, C is D301 macroporous basic resin, D201 macroporous basic resin, E is 001 × 7Na732
The ordinate of fig. 1-3 is the caffeic acid content of the product.
FIG. 4 is a high performance liquid chromatography assay of caffeic acid of example 3.
Detailed Description
The present invention will be described in detail with reference to examples. It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict.
Example 1
Weighing 500g of dried rosemary, adding 10L of ethanol (30%, 40%, 50%, 60%, 70% and 80%), refluxing and extracting for 2 hours, filtering, concentrating the filtrate under reduced pressure to Brix 40-60%, standing, cooling, adding 50g of glucose, and mixing uniformly. Adding 5mL of mixed bacteria liquid of 70% of lactic acid bacteria and 30% of yeast into the concentrated solution, uniformly mixing, controlling the temperature at 37 ℃, carrying out heat preservation culture for 7 days, adding water into the obtained liquid until Brix 10-12%, adjusting the pH value to 4-6 by using dilute hydrochloric acid, and centrifuging to obtain clear centrifugal liquid; passing the centrifuged clear solution through NKA-9 macroporous adsorption resin; and after the column is loaded, eluting by pure water to remove impurities which are not adsorbed by the resin, desorbing by using 50-60% ethanol solution with PH 4-6, starting to receive liquid when the effluent is weakly acidic, washing to be neutral by using deionized water after the caffeic acid solution is loaded, stopping water loading and liquid receiving, concentrating under reduced pressure to Brix 30-35%, decolorizing and filtering the concentrated solution by using 325-mesh active carbon filter cloth, concentrating again, drying in vacuum, and detecting the caffeic acid content.
As shown in the result of figure 1, when the ethanol concentration is 30% -40%, the caffeic acid content is 22.1% -42.7%, and the caffeic acid content gradually increases along with the increase of the proportion of the ethanol. At an ethanol concentration of 60, the caffeic acid content was 57.1%, reaching a maximum value, which then decreased with increasing ethanol concentration.
Therefore, the ethanol concentration is preferably 50% -60% for extraction.
Example 2
Weighing 500g of dried rosemary pieces, adding 10L of 60-65% ethanol, performing reflux extraction for 2 hours, filtering, concentrating the filtrate under reduced pressure to Brix 40-60%, standing, cooling, adding 50g of glucose, and uniformly mixing. Mixing the lactic acid bacteria: adding 5mL of mixed yeast solution with the yeast ratio of (A1: 9, B3:7, C5: 5, D7: 3 and E9: 1) into the concentrated solution, uniformly mixing, controlling the temperature at 37 ℃, carrying out heat preservation culture for 7 days, adding water into the obtained liquid to Brix 10-12%, adjusting the pH value to 4-6 by using dilute hydrochloric acid, and centrifuging to obtain clear centrifugal solution; passing the centrifuged clear solution through NKA-9 macroporous adsorption resin; and after the column is loaded, eluting by pure water to remove impurities which are not adsorbed by the resin, desorbing by using 50-60% ethanol solution with PH 4-6, starting to receive liquid when the effluent is weakly acidic, washing to be neutral by using deionized water after the caffeic acid solution is loaded, stopping water loading and liquid receiving, concentrating under reduced pressure to Brix 30-35%, decolorizing and filtering the concentrated solution by using 325-mesh active carbon filter cloth, concentrating again, drying in vacuum, and detecting the caffeic acid content.
As shown in the results of fig. 2, lactic acid bacteria: when the yeast ratio is 1:9, the caffeic acid content is only 14.8%, and the caffeic acid content gradually increases with the increase of the proportion of the lactic acid bacteria. In the case of lactic acid bacteria: when the yeast ratio is 7:3, the caffeic acid content is 57.4%, the highest value is reached, and then the caffeic acid content is reduced along with the increase of lactic acid bacteria.
Therefore, the mixed bacteria liquid with 70% of lactic acid bacteria and 30% of yeast is preferably fermented.
Example 2
Weighing 500g of dried rosemary pieces, adding 10L of 60-65% ethanol, performing reflux extraction for 2 hours, filtering, concentrating the filtrate under reduced pressure to Brix 40-60%, standing, cooling, adding 50g of glucose, and uniformly mixing. Adding 5mL of mixed bacteria liquid containing 70% of lactic acid bacteria and 30% of yeast into the concentrated solution, uniformly mixing, controlling the temperature at 37 ℃, carrying out heat preservation culture for 7 days, adding water into the obtained liquid to Brix 10-12%, adjusting the pH value to 4-6 by using dilute hydrochloric acid, and centrifuging to obtain clear centrifugal liquid; passing the supernatant through macroporous adsorbent resin (NKA-9 macroporous resin, D1O1 macroporous resin, C D301 macroporous alkaline resin, D201 macroporous alkaline resin, E001 × 7Na 732); and after the column is loaded, eluting by pure water to remove impurities which are not adsorbed by the resin, desorbing by using 50-60% ethanol solution with PH 4-6, starting to receive liquid when the effluent is weakly acidic, washing to be neutral by using deionized water after the caffeic acid solution is loaded, stopping water loading and liquid receiving, concentrating under reduced pressure to Brix 30-35%, decolorizing and filtering the concentrated solution by using 325-mesh active carbon filter cloth, concentrating again, drying in vacuum, and detecting the caffeic acid content.
As shown in the results of fig. 3, the effect of different adsorption resins on the total coumarin content was very significant. The adsorption resin is D1O1, and the total coumarin content is only 12.6%. The adsorption resin is D301 and NKA-9, and the total coumarin content can reach 42.4% and 58.4%.
Therefore, medium polarity NKA-9 macroporous resins are preferred.
Example 3
Weighing 10kg of dried rosemary pieces, putting into a 200L extraction tank, adding 120L of 60% -65% ethanol, and stirring; and opening a jacket, heating with steam to 80-83 ℃, circulating for 2 hours, then starting normal percolation, collecting the percolate until the Brix value is less than 1%, stopping collection, filtering, concentrating the filtrate under reduced pressure to Brix 40-60%, standing, cooling, adding 1kg of glucose, and uniformly mixing. Adding 10mL of mixed solution of 70% of lactic acid bacteria and 30% of yeast into the concentrated solution, uniformly mixing, controlling the temperature at 37 ℃, carrying out heat preservation culture for 7 days, adding water into the obtained liquid to Brix 10-12%, adjusting the pH value to 4-6 by using dilute hydrochloric acid, and centrifuging to obtain clear centrifugal solution; passing the centrifugated clear liquid through 25L NKA-9 macroporous adsorbent resin at a flow rate of 25L/h; after the column loading is finished, 150L of pure water is used for eluting impurities which are not adsorbed by the resin at the flow rate of 50L/h, then 25L of 50-60% ethanol solution with the pH of 4-6 is used for desorbing at the flow rate of 10L/h, and when the effluent liquid is weakly acidic, the liquid begins to be received. And after the caffeic acid solution is applied, washing the caffeic acid solution to be neutral by deionized water, stopping applying water and liquid receiving, concentrating the solution under reduced pressure to Brix 30-35%, decoloring and filtering the concentrated solution by 325-mesh active carbon filter cloth, concentrating the filtrate again until the solid content reaches more than 50%, and drying the concentrated solution in vacuum to obtain 462g of dried block with the caffeic acid content of 57.3%. Liquid chromatography data for caffeic acid is shown in fig. 4A.
Example 4
Weighing 300kg of dried rosemary pieces, putting into a 5000L extraction tank, adding 3000L of 60-65% ethanol, and stirring; and opening a jacket, heating with steam to 80-83 ℃, circulating for 2 hours, then starting normal percolation, collecting the percolate until the Brix value is less than 1%, stopping collection, filtering, concentrating the filtrate under reduced pressure to Brix 40-60%, standing, cooling, adding 30kg of glucose, and uniformly mixing. Adding 300mL of mixed solution of 70% of lactic acid bacteria and 40% of yeast into the concentrated solution, uniformly mixing, controlling the temperature at 37 ℃, carrying out heat preservation culture for 7 days, adding water into the obtained liquid to Brix 10-12%, adjusting the pH value to 4-6 by using dilute hydrochloric acid, and centrifuging to obtain clear centrifugal solution; passing the centrifuged clear liquid through 1000L NKA-9 macroporous adsorption resin at the flow rate of 1000L/h; after the column loading is finished, eluting with 3000L of pure water at a flow rate of 2000L/h to remove impurities not adsorbed by the resin, desorbing with 1500L of 50-60% ethanol solution with pH of 4-6 at a flow rate of 500L/h, and starting to receive liquid when the effluent is weakly acidic. And after the caffeic acid solution is applied, washing the caffeic acid solution to be neutral by deionized water, stopping applying water and liquid receiving, concentrating the solution under reduced pressure to Brix 30-35%, decoloring and filtering the concentrated solution by 325-mesh active carbon filter cloth, concentrating the filtrate again until the solid content reaches more than 50%, and drying the concentrated solution in vacuum to obtain 13.27kg of dried blocks with the caffeic acid content of 58.1%. Liquid chromatography data for caffeic acid is shown in fig. 4B. After amplified extraction, the extraction rate of the total coumarin is also 58.1 percent, the total coumarin is consistent with the small test level, and meanwhile, the chromatogram shows that the product composition and the content are highly consistent, so that the caffeic acid content in industrial production is greatly improved, and the industrial application prospect is excellent.
The foregoing examples are set forth to illustrate the present invention more clearly and are not to be construed as limiting the scope of the invention, which is defined in the appended claims to which the invention pertains, as modified in all equivalent forms, by those skilled in the art after reading the present invention.
Claims (10)
1. A method for extracting caffeic acid from rosemary is characterized by comprising the following steps:
s1, crushing rosemary, extracting with ethanol at 80-83 ℃, and carrying out solid-liquid separation on the extracting solution to obtain a clear extracting solution;
s2, concentrating the clear extracting solution obtained in the step S1 to Brix 40-60% to obtain a concentrated solution, and adding a carbon source;
s3, inoculating zymophyte into the concentrated solution added with the carbon source in the step S2, adjusting the pH value to 4-6, and fermenting to obtain caffeic acid liquid;
s4, adding water into the caffeic acid liquid obtained in the step S3 to Brix 11-12%, adjusting the pH value to 4-6, and performing solid-liquid separation to obtain clear supernatant;
s5, adding the supernatant obtained in the step S4 into macroporous adsorption resin, and washing the macroporous adsorption resin with water to remove unadsorbed impurities; washing the macroporous adsorption resin with desorption solution with the pH value of 4-6, collecting the washed desorption solution, and concentrating under reduced pressure to Brix 30-35%;
s6, decolorizing and filtering the concentrated solution, concentrating the filtrate again, and drying to obtain the caffeic acid extract.
2. The method according to claim 1, wherein in step S1 rosemary is crushed to a diameter of 0.5cm or less and the extraction is a percolation extraction.
3. The method according to claim 1, wherein the ethanol concentration in the step S1 is 50% -60%.
4. The method of claim 1, wherein the carbon source is glucose; the fermentation temperature is 35-37 ℃ and the period is 7-10 days; the solid-liquid separation is filtration or centrifugation.
5. The method according to claim 1, wherein the fermentation tubes comprise 10-90% by mass of lactic acid bacteria and 10-90% by mass of yeast.
6. The method according to claim 1, wherein the fermentation tubes comprise 60-80% of lactic acid bacteria and 20-40% of yeast by mass.
7. The method according to claim 1, wherein the fermentation tubes comprise 70% of lactic acid bacteria and 30% of yeast by mass.
8. The method of claim 1, wherein the macroporous adsorbent resin is NKA-9 macroporous resin, D1O1 macroporous resin, D301 macroporous basic resin, D201 macroporous basic resin, 001 x 7Na 732.
9. The method of claim 1, wherein the macroporous adsorbent resin is a medium polarity macroporous adsorbent resin; preferably, the macroporous adsorption resin is NKA-9 macroporous adsorption resin.
10. The method according to claim 1, wherein the desorption solution is an ethanol solution with a concentration of 50-60% by volume; the active carbon filter cloth for decolorization and filtration has 325 meshes; the drying is vacuum drying or spray drying until the water content of the caffeic acid extract is not more than 5%.
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| 曹姗、祖元刚、张琳: "LC-MS/MS方法同时检测超声提取迷迭香(Rosmarnus officinalis L.)叶片中的4种主要成分" * |
| 王珲: "迷迭香化学成分与质量评价研究" * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113173848A (en) * | 2021-05-12 | 2021-07-27 | 南京合谷生命生物科技有限公司 | Method for extracting and separating caffeic acid from fermentation liquor |
| CN113173848B (en) * | 2021-05-12 | 2022-02-18 | 南京合谷生命生物科技有限公司 | Method for extracting and separating caffeic acid from fermentation liquor |
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| CN111676264B (en) | 2023-05-16 |
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