CN114736123B - Method for extracting high-purity rosmarinic acid - Google Patents
Method for extracting high-purity rosmarinic acid Download PDFInfo
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Abstract
The invention discloses an extraction method of high-purity rosmarinic acid, which comprises a plurality of extraction, precipitation and filtration steps, particularly, a specific-aperture filtration device is selected for filtration in the step (4), a two-aqueous-phase system is selected for extraction in the step (6), and impurities which are difficult to separate in the rosmarinic acid are effectively removed, and meanwhile, the loss of the rosmarinic acid in the extraction process is reduced, so that the purity of the rosmarinic acid is ensured, the yield of the high-purity rosmarinic acid is improved, and the industrial production and application of the rosmarinic acid are facilitated.
Description
Technical Field
The invention relates to the technical field of plant component extraction, in particular to a method for extracting high-purity rosmarinic acid.
Background
Rosmarinic acid (Rosa for short) belongs to diterpenoids in terpenes, is a water-soluble natural phenolic acid compound separated from rosemary of Labiatae, has wide distribution, and mainly exists in various plants of Labiatae, boraginaceae, cucurbitaceae, tilia, umbelliferae, especially in Labiatae and Arnebiae. Rosmarinic acid is a natural antioxidant with stronger antioxidant activity than vitamin E, caffeic acid, chlorogenic acid, folic acid and the like, and can reduce CCl 4-induced liver injury by scavenging free radicals, reducing peroxide or peroxynitrite, promoting protein synthesis, regulating fat metabolism, inhibiting collagen synthesis, promoting degradation and the like. Rosmarinic acid also has strong anti-inflammatory activity, has remarkable antibacterial effect on staphylococcus aureus, escherichia coli and salmonella, and has strong antibacterial effect on gram-positive bacteria. At present, rosmarinic acid has shown important application value in the fields of pharmacy, food, cosmetics and the like.
At present, research on the extraction process of rosmarinic acid is a hot spot of food industry and plant extraction industry, but the processes for preparing the product with high rosmarinic acid content are very few, and most of the processes use a macroporous resin adsorption separation method, and the method has the defects of low production efficiency, high product solvent residue, serious environmental pollution and the like; meanwhile, the partial preparation method has the defect of low yield of the high-content rosemary product, so that the production income is low, and the large-scale production and application of rosmarinic acid are not facilitated.
Disclosure of Invention
The invention aims to overcome the defect of low product yield of the existing preparation method of high-purity rosmarinic acid, and provides an extraction method of high-purity rosmarinic acid; in the extraction process of the high-purity rosmarinic acid, the filtration equipment with specific aperture is selected for filtration in the step (4), and the two-aqueous phase system is selected for extraction in the step (6), so that the loss of the rosmarinic acid in the extraction process is reduced while the impurities which are difficult to separate in the rosmarinic acid are effectively removed, the purity of the rosmarinic acid is ensured, the yield of the high-purity rosmarinic acid is improved, and the industrial production and application of the rosmarinic acid are facilitated.
In order to achieve the above object, the present invention provides a method for extracting high-purity rosmarinic acid, comprising the steps of:
(1) Steam distilling and extracting fresh rosemary leaves, and removing distillate to obtain distilled rosemary leaves;
(2) Adding water with the weight of She Tongdeng to distilled rosemary leaves for water extraction for 1-2 times, and combining water extracts to obtain primary extract A;
(3) Concentrating the primary extract A under reduced pressure, controlling the mass percentage of solute in the concentrated solution to be 5% -10%, adding acid liquor to adjust the pH value to 2-4, standing for layering, and taking supernatant;
(4) Filtering the supernatant with a filter membrane with a pore diameter of 0.2-0.5 μm to obtain filtrate B;
(5) Adding n-butanol into the filtrate B, standing for layering after extraction treatment, repeatedly extracting for 2-4 times, mixing n-butanol extract, concentrating under reduced pressure to 1/20-1/25 of the weight of n-butanol, cooling, standing, and collecting precipitate A;
(6) Dissolving the precipitate A with a 20-40% dihydrogen phosphate solution with the mass percentage of 5-10 volume times of the mass of the precipitate A at the temperature of 25-50 ℃, filtering, adding an ethanol solution with the volume percentage of 95% into the filtrate, stirring and extracting for 15-25min, and standing to separate an upper layer solution C; wherein the volume ratio of the ethanol solution to the dihydrogen phosphate solution is 4:6;
(7) Adding acid into the upper layer solution C to adjust the pH value to 3-4, concentrating under reduced pressure until the solution is saturated, standing for crystallization to obtain a precipitate B;
(8) Adding the precipitate B obtained in the step 7 into acetic acid, heating for dissolution, filtering, cooling, standing for recrystallization, and repeating for several times to obtain a precipitate C;
(9) Adding water into the precipitate C, heating for dissolving, cooling, standing for recrystallization, and repeating for several times to obtain high-purity rosmarinic acid.
In the step (2), preferably, the water is used as an extraction solvent, the extraction temperature is 60-70 ℃, the extraction time is 40-50min, and the preferred water extraction process and parameters have no fat-soluble rosemary extract component, so that the complexity of subsequent purification is reduced;
wherein in the step (3), the temperature of the reduced pressure concentration is 60-70 ℃ and the pressure is-0.07 MPa to-0.09 MPa; the acid liquor is one of phosphoric acid, hydrochloric acid or citric acid. The preferred concentration conditions have higher concentration efficiency and lower energy consumption. Standing for 24-48h; the preferable acid liquid type has better impurity removing effect and higher yield.
Wherein in the step (4), preferably, the pore diameter of the filtering membrane is 0.2-0.3 μm; the pore diameter of the filtering membrane is preferable, and most of impurity substances with molecular weight larger than rosmarinic acid can be removed by filtering, so that the purification treatment in the later stage is facilitated, and the yield of high-purity rosmarinic acid can be improved.
In the step (5), the dosage of the n-butanol is preferably 1-1.5 times of the weight of the filtrate B, and the extraction time is 20-30min; the reduced pressure concentration temperature is 65-75 ℃, and the pressure is-0.08 MPa to-0.1 MPa; standing for 12-24h; the optimized extraction and concentration conditions can better extract impurities, thereby being beneficial to the later purification treatment and improving the yield of the high-purity rosmarinic acid.
In the step (6), preferably, the monobasic phosphate solution is one or two of a monobasic sodium phosphate solution and a monobasic potassium phosphate solution; the application of the double aqueous phase system of the dihydrogen phosphate solution and the ethanol solution better separates impurities from active ingredients, and is more beneficial to the use of crystallization technology in the subsequent step;
in the step (7), preferably, the acid liquid is phosphoric acid, hydrochloric acid or citric acid, and is concentrated under reduced pressure in an acidic environment, so that the active ingredients are protected more easily;
in the step (8), preferably, the adding amount of the acetic acid is 2-3 times of the weight of the filter residue, and the standing time is 12-24 hours; the repetition times are 1-2 times; the preferred conditions are better able to remove the impurities from rosmarinic acid.
Wherein, in the steps (8) and (9), the temperature of heating and dissolving is 60-70 ℃, and the preferable dissolving temperature can better remove the impurities in the rosmarinic acid.
Wherein in the step (9), the water is preferably added in an amount of 1.5 to 2.5 times the weight of the precipitate C, and the standing time is 12 to 48 hours; the repetition times are 2-3 times; the preferred conditions are better able to remove the impurities from rosmarinic acid.
Compared with the prior art, the invention has the beneficial effects that:
1. according to the extraction method, the filtering equipment with the specific aperture is selected for filtering, and the impurity in the rosmarinic acid is removed through the two-aqueous-phase system, so that the loss of the rosmarinic acid in the extraction process is reduced while the impurity in the rosmarinic acid is effectively removed, and the yield of the high-purity rosmarinic acid is improved while the high purity of the rosmarinic acid is ensured.
2. The extraction method is simple and reliable, can obviously improve the yield of the rosmarinic acid, and is beneficial to the large-scale production and application of the high-purity rosmarinic acid.
3. The extraction method has the advantages of good safety, recyclable solvent, low production cost, less production waste and less environmental pollution.
Detailed Description
The present invention will be described in further detail with reference to test examples and specific embodiments, but it should not be construed that the scope of the above subject matter of the present invention is limited to the following examples, and all techniques realized based on the present invention are within the scope of the present invention.
Example 1:
an extraction method of high-purity rosmarinic acid comprises the following steps:
(1) Putting 100KG of fresh rosemary leaves into an extraction tank, distilling with steam for 50min, and removing distillate to obtain distilled rosemary leaves;
(2) Adding 100KG water into distilled rosemary leaves, extracting the distilled rosemary leaves for 2 times at 65 ℃ for 50min, and combining the extracting solutions to obtain primary extract A;
(3) Concentrating the primary extract A to 30KG under reduced pressure, wherein the solute percentage in the solution is 10%, adding hydrochloric acid to adjust the pH value to 3, standing and layering for 36h, and taking supernatant;
(4) Filtering the supernatant with a filter membrane with the pore diameter of 0.3 μm to obtain filtrate B;
(5) Adding 45KG n-butanol into the filtrate B, extracting for 30min, standing for layering for 1h after extraction treatment, extracting for 3 times repeatedly, mixing n-butanol extract, concentrating under reduced pressure to 2KG under the condition that the pressure is-0.08 MPa at 65 ℃, cooling, standing, and collecting precipitate A;
(6) Dissolving the precipitate A with 18L of 28% potassium dihydrogen phosphate solution at 35 deg.C; filtering, adding 95% ethanol solution with volume of 12L into the filtrate, stirring and extracting for 20min, standing and separating for 1h, and separating out upper layer solution C;
(7) Adding hydrochloric acid into the solution C to adjust the pH value to 3, concentrating under reduced pressure until the solution just has insoluble substances separated out, standing for crystallization to obtain a precipitate B;
(8) Adding the precipitate B obtained in the step 7 into 1L of acetic acid, heating to 65 ℃ for dissolution, filtering while the precipitate B is hot, cooling the filtrate, standing for recrystallization, and repeating the steps for 2 times to obtain a precipitate C;
(9) Adding water 0.5L into the precipitate C, heating to 65deg.C for dissolving, cooling, standing for recrystallization, and repeating for 2 times to obtain high-purity rosmarinic acid.
Example 2:
an extraction method of high-purity rosmarinic acid comprises the following steps:
(1) Putting 100KG of fresh rosemary leaves into an extraction tank, distilling with steam for 50min, and removing distillate to obtain distilled rosemary leaves;
(2) Adding 100KG water into distilled rosemary leaves, extracting the distilled rosemary leaves for 2 times at 60 ℃ for 45min, and combining the extracting solutions to obtain primary extract A;
(3) Concentrating the primary extract A to 37.5KG under reduced pressure, wherein the solute percentage in the solution is 8.2%, adding hydrochloric acid to adjust the pH value to 3, standing and layering for 24 hours, and taking supernatant;
(4) Filtering the supernatant with a filter membrane with the pore diameter of 0.4 μm to obtain filtrate B;
(5) Adding 40KG of n-butanol into the filtrate B, extracting for 30min, standing for layering for 1h after extraction treatment, extracting for 4 times repeatedly, mixing n-butanol extract, concentrating under reduced pressure to 2KG under the condition that the pressure is-0.08 MPa at 70 ℃, cooling, standing, and collecting precipitate A;
(6) Dissolving the precipitate A with 15L of 32% potassium dihydrogen phosphate solution at 40deg.C; filtering, adding 95% ethanol solution with volume of 10L into the filtrate, stirring and extracting for 20min, standing and separating for 1h, and separating out upper layer solution C;
(7) Adding phosphoric acid into the solution C to adjust the pH value to 3, concentrating under reduced pressure until the solution just has insoluble substances separated out, standing for crystallization to obtain a precipitate B;
(8) Adding 1L of acetic acid into the precipitate B obtained in the step 7, heating to 60 ℃ for dissolution, filtering while the precipitate B is hot, cooling the filtrate, standing for recrystallization, and repeating for 1 time to obtain a precipitate C;
(9) Adding water into precipitate C0.6L, heating to 60deg.C for dissolving, cooling, standing for recrystallization, and repeating for 3 times to obtain high-purity rosmarinic acid.
Example 3:
an extraction method of high-purity rosmarinic acid comprises the following steps:
(1) Putting 100KG of fresh rosemary leaves into an extraction tank, distilling with steam for 50min, and removing distillate to obtain distilled rosemary leaves;
(2) Adding 100KG water into distilled rosemary leaves, extracting the distilled rosemary leaves for 1 time at 70 ℃ for 50min to obtain primary extract A;
(3) Concentrating the primary extract A to 25KG under reduced pressure, wherein the solute percentage in the solution is 9%, adding hydrochloric acid to adjust the pH value to 2, standing and layering for 30h, and taking supernatant;
(4) Filtering the supernatant with a filter membrane with the pore diameter of 0.5 μm to obtain filtrate B;
(5) Adding 33KG of n-butanol into the filtrate B, extracting for 30min, standing for layering for 1h after extraction treatment, extracting for 3 times repeatedly, mixing n-butanol extract, concentrating under reduced pressure to 1.5KG under the condition that the pressure is-0.08 MPa at 65 ℃, cooling, standing, and collecting precipitate A;
(6) Dissolving the precipitate A with 12L of 30% sodium dihydrogen phosphate solution at 40deg.C; filtering, adding 95% ethanol solution with volume of 8L into the filtrate, stirring and extracting for 20min, standing and separating for 1h, and separating out upper layer solution C;
(7) Adding phosphoric acid into the solution C to adjust the pH value to 3, concentrating under reduced pressure until the solution just has insoluble substances separated out, standing for crystallization to obtain a precipitate B;
(8) Adding 0.8L of acetic acid into the precipitate B obtained in the step 7, heating to 70 ℃ for dissolution, filtering while the precipitate B is hot, cooling the filtrate, standing for recrystallization, and repeating the steps for 2 times to obtain a precipitate C;
(9) Adding water 0.4L into the precipitate C, heating to 70deg.C for dissolving, cooling, standing for recrystallization, and repeating for 3 times to obtain high-purity rosmarinic acid.
Comparative example 1:
an extraction method of high-purity rosmarinic acid comprises the following steps:
(1) Putting 100KG of fresh rosemary leaves into an extraction tank, distilling with steam for 50min, and removing distillate to obtain distilled rosemary leaves;
(2) Adding 100KG water into distilled rosemary leaves, extracting the distilled rosemary leaves for 2 times at 65 ℃ for 50min, and combining the extracting solutions to obtain primary extract A;
(3) Concentrating the primary extract A to 30KG under reduced pressure, wherein the solute percentage in the solution is 10%, adding hydrochloric acid to adjust the pH value to 3, standing and layering for 36h, and taking supernatant;
(4) Adding 45KG n-butanol into the supernatant for 30min, standing for layering for 1h after extraction treatment, extracting for 3 times repeatedly, mixing n-butanol extractive solutions, concentrating under reduced pressure to 2KG at 65 deg.C and under-0.08 MPa, cooling, standing, and collecting precipitate A;
(5) Dissolving the precipitate A with 18L of 28% potassium dihydrogen phosphate solution at 35 deg.C; filtering, adding 95% ethanol solution with volume of 12L into the filtrate, stirring and extracting for 20min, standing and separating for 1h, and separating out upper layer solution C;
(6) Adding hydrochloric acid into the solution C to adjust the pH value to 3, concentrating under reduced pressure until the solution just has insoluble substances separated out, standing for crystallization to obtain a precipitate B;
(7) Adding the precipitate B obtained in the step 6 into 1L of acetic acid, heating to 65 ℃ for dissolution, filtering while the precipitate B is hot, cooling the filtrate, standing for recrystallization, and repeating the steps for 2 times to obtain a precipitate C;
(8) Adding water 0.5L into the precipitate C, heating to 65deg.C for dissolving, cooling, standing for recrystallization, and repeating for 2 times to obtain high-purity rosmarinic acid.
Comparative example 2:
an extraction method of high-purity rosmarinic acid comprises the following steps:
(1) Putting 100KG of fresh rosemary leaves into an extraction tank, distilling with steam for 50min, and removing distillate to obtain distilled rosemary leaves;
(2) Adding 100KG water into distilled rosemary leaves, extracting the distilled rosemary leaves for 2 times at 65 ℃ for 50min, and combining the extracting solutions to obtain primary extract A;
(3) Concentrating the primary extract A to 30KG under reduced pressure, wherein the solute percentage in the solution is 10%, adding hydrochloric acid to adjust the pH value to 3, standing and layering for 36h, and taking supernatant;
(4) Filtering the supernatant with a filter membrane with the pore diameter of 0.3 μm to obtain filtrate B;
(5) Adding 45KG of n-butanol into the filtrate B, extracting for 30min, standing for layering for 1h after extraction treatment, extracting for 3 times repeatedly, combining n-butanol extract, concentrating under reduced pressure to 5KG under the condition that the pressure is-0.08 MPa at 65 ℃, cooling, standing, and collecting precipitate A;
(6) Dissolving the precipitate A with 18L of 28% potassium dihydrogen phosphate solution at 35 deg.C; filtering, adding 95% ethanol solution with volume of 12L into the filtrate, stirring and extracting for 20min, standing and separating for 1h, and separating out upper layer solution C;
(7) Adding hydrochloric acid into the solution C to adjust the pH value to 3, concentrating under reduced pressure until the solution just has insoluble substances separated out, standing for crystallization to obtain a precipitate B;
(8) Adding the precipitate B obtained in the step 7 into 1L of acetic acid, heating to 65 ℃ for dissolution, filtering while the precipitate B is hot, cooling the filtrate, standing for recrystallization, and repeating the steps for 2 times to obtain a precipitate C;
(9) Adding water 0.5L into the precipitate C, heating to 65deg.C for dissolving, cooling, standing for recrystallization, and repeating for 2 times to obtain high-purity rosmarinic acid.
Comparative example 3:
an extraction method of high-purity rosmarinic acid comprises the following steps:
(1) Putting 100KG of fresh rosemary leaves into an extraction tank, distilling with steam for 50min, and removing distillate to obtain distilled rosemary leaves;
(2) Adding 100KG water into distilled rosemary leaves, extracting the distilled rosemary leaves for 2 times at 65 ℃ for 50min, and combining the extracting solutions to obtain primary extract A;
(3) Concentrating the primary extract A to 30KG under reduced pressure, wherein the solute percentage in the solution is 10%, adding hydrochloric acid to adjust the pH value to 3, standing and layering for 36h, and taking supernatant;
(4) Filtering the supernatant with a filter membrane with the pore diameter of 0.3 μm to obtain filtrate B;
(5) Adding 45KG n-butanol into the filtrate B, extracting for 30min, standing for layering for 1h after extraction treatment, extracting for 3 times repeatedly, mixing n-butanol extract, concentrating under reduced pressure to 2KG under the condition that the pressure is-0.08 MPa at 65 ℃, cooling, standing, and collecting precipitate A;
(6) Dissolving the precipitate A with 18L of 15% potassium dihydrogen phosphate solution at 35 deg.C; filtering, adding 95% ethanol solution with volume of 12L into the filtrate, stirring and extracting for 20min, standing and separating for 1h, and separating out upper layer solution C;
(7) Adding hydrochloric acid into the solution C to adjust the pH value to 3, concentrating under reduced pressure until the solution just has insoluble substances separated out, standing for crystallization to obtain a precipitate B;
(8) Adding the precipitate B obtained in the step 7 into 1L of acetic acid, heating to 65 ℃ for dissolution, filtering while the precipitate B is hot, cooling the filtrate, standing for recrystallization, and repeating the steps for 2 times to obtain a precipitate C;
(9) Adding water 0.5L into the precipitate C, heating to 65deg.C for dissolving, cooling, standing for recrystallization, and repeating for 2 times to obtain high-purity rosmarinic acid.
Comparative example 4:
an extraction method of high-purity rosmarinic acid comprises the following steps:
(1) Putting 100KG of fresh rosemary leaves into an extraction tank, distilling with steam for 50min, and removing distillate to obtain distilled rosemary leaves;
(2) Adding 100KG water into distilled rosemary leaves, extracting the distilled rosemary leaves for 2 times at 65 ℃ for 50min, and combining the extracting solutions to obtain primary extract A;
(3) Concentrating the primary extract A to 30KG under reduced pressure, wherein the solute percentage in the solution is 10%, adding hydrochloric acid to adjust the pH value to 3, standing and layering for 36h, and taking supernatant;
(4) Filtering the supernatant with a filter membrane with the pore diameter of 0.3 μm to obtain filtrate B;
(5) Adding 45KG n-butanol into the filtrate B, extracting for 30min, standing for layering for 1h after extraction treatment, extracting for 3 times repeatedly, mixing n-butanol extract, concentrating under reduced pressure to 2KG under the condition that the pressure is-0.08 MPa at 65 ℃, cooling, standing, and collecting precipitate A;
(6) Dissolving the precipitate A with 18L of 28% potassium dihydrogen phosphate solution at 35 deg.C; filtering, adding 95% ethanol solution with volume of 12L into the filtrate, stirring and extracting for 20min, standing and separating for 1h, and separating out upper layer solution C;
(7) Adding hydrochloric acid into the solution C to adjust the pH value to 3, concentrating under reduced pressure until the solution just has insoluble substances separated out, standing for crystallization to obtain a precipitate B;
(8) Adding the precipitate B obtained in the step 7 into 1L of acetic acid, heating to 65 ℃ for dissolution, filtering while the precipitate B is hot, cooling the filtrate, standing for recrystallization, and repeating the steps for 2 times to obtain a precipitate C; the obtained precipitate C is high-purity rosmarinic acid.
Experimental example:
concentration measurement: the purity of rosmarinic acid (rosmarinic acid content/total mass% of crystals) was determined by liquid chromatography.
And (3) measuring the weight of the crystal: analytical balance
The weight of the rosemary material (fresh rosemary leaves) in each embodiment of the invention is 100kg.
Test results:
| project | Purity (%) | Weight (g) |
| Example 1 | 99.4 | 119.5 |
| Example 2 | 99.3 | 116.4 |
| Example 3 | 99.4 | 112.8 |
| Comparative example 1 | 80.2 | 146.6 |
| Comparative example 2 | 98.4 | 61.2 |
| Comparative example 3 | 75.6 | 158.2 |
| Comparative example 4 | 94.1 | 124.1 |
Conclusion of experiment: the preparation method in the embodiments 1-3 can effectively improve the yield of rosmarinic acid and reduce the impurity content; in comparative example 1, the microporous membrane filtration process in step (4) is not adopted, and a large amount of impurities enter the subsequent purification process, so that the content of rosmarinic acid is obviously reduced; in comparative example 2, step (5) resulted in a significant decrease in the final rosmarinic acid yield due to insufficient concentration under reduced pressure, a large amount of solution, and a decrease in the precipitates; in the step (6) of the comparative example 3, the concentration of the potassium dihydrogen phosphate solution is too low to effectively separate impurities, so that the purity of rosmarinic acid obtained by subsequent crystallization is obviously reduced; in comparative example 4, the step (9) was not continued to recrystallize with water recrystallization, and the content of rosmarinic acid obtained was only 94.1%. The experimental results show that the pore diameter, the extraction system and the recrystallization solvent selected by filtering in the preparation process of the high-purity rosmarinic acid have direct influence on the content and the yield of the final product.
Claims (8)
1. The method for extracting rosmarinic acid is characterized by comprising the following steps:
(1) Steam distilling and extracting fresh rosemary leaves, and removing distillate to obtain distilled rosemary leaves;
(2) Adding water with the weight of She Tongdeng to distilled rosemary leaves for water extraction for 1-2 times, and combining water extracts to obtain primary extract A;
(3) Concentrating the primary extract A under reduced pressure, controlling the mass percentage of solute in the concentrated solution to be 5% -10%, adding acid to adjust the pH value to 2-4, standing for layering, and taking supernatant;
(4) Filtering the supernatant with a filter membrane with a pore diameter of 0.2-0.5 μm to obtain filtrate B;
(5) Adding n-butanol into the filtrate B, standing for layering after extraction treatment, repeatedly extracting for 2-4 times, mixing n-butanol extract, concentrating under reduced pressure to 1/20-1/25 of the weight of n-butanol, cooling, standing, and collecting precipitate A;
(6) Dissolving the precipitate A with a solution of 28-40% of dihydrogen phosphate with the mass percent of 5-10 volume times of the mass of the precipitate A at the temperature of 25-50 ℃, filtering, adding an ethanol solution with the volume percent of 95% into the filtrate, stirring and extracting for 15-25min, and standing to separate an upper layer solution C; wherein the volume ratio of the ethanol solution to the dihydrogen phosphate solution is 4:6;
(7) Adding acid into the upper layer solution C to adjust the pH value to 3-4, concentrating under reduced pressure until the solution is saturated, standing for crystallization to obtain a precipitate B;
(8) Adding the precipitate B obtained in the step 7 into acetic acid, heating for dissolution, filtering, cooling, standing for recrystallization, and repeating for several times to obtain a precipitate C;
(9) Adding water into the precipitate C, heating for dissolving, cooling, standing for recrystallization, and repeating for several times to obtain crystal of rosmarinic acid;
wherein, the monobasic phosphate solution in the step (6) is one or two of sodium monobasic phosphate solution and potassium monobasic phosphate solution; the acid in the step (3) and the step (7) is one of phosphoric acid, hydrochloric acid or citric acid.
2. The method according to claim 1, wherein in the step (2), the water extraction temperature is 60-70 ℃ and the water extraction time is 40-50min.
3. The method according to claim 1, wherein in the step (3), the reduced pressure concentration is performed at a temperature of 60 to 70 ℃ and a pressure of-0.07 to-0.09 MPa.
4. The method according to claim 1, wherein in the step (4), the pore size of the filtration membrane is 0.2 to 0.3. Mu.m.
5. The method according to claim 1, wherein in the step (5), the n-butanol is used in an amount of 1 to 1.5 times the weight of the filtrate B.
6. The method according to claim 1, wherein in the step (8), the amount of acetic acid is 2 to 3 times the weight of the precipitate B.
7. The method for extracting rosmarinic acid as claimed in claim 1, wherein in the steps (8) and (9), the temperature of the heating dissolution is 60 to 70 ℃.
8. The method for extracting rosmarinic acid as claimed in claim 1, wherein in the step (9), the amount of water is 1.5 to 2.5 times by weight of the precipitate C.
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| JPS59132894A (en) * | 1982-12-23 | 1984-07-31 | ア−・ナツタ−マン・ウント・コムパニ−・ゲ−エムベ−ハ− | Production of rosmarine |
| CN1995007A (en) * | 2005-12-26 | 2007-07-11 | 山东绿叶天然药物研究开发有限公司 | Process for preparing rosmarinic acid |
| CN106542999A (en) * | 2016-10-14 | 2017-03-29 | 无锡加莱克色谱科技有限公司 | A kind of purification process of rosmarinic acid |
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