CN113461531A - Method for extracting rosmarinic acid - Google Patents
Method for extracting rosmarinic acid Download PDFInfo
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- CN113461531A CN113461531A CN202110668807.2A CN202110668807A CN113461531A CN 113461531 A CN113461531 A CN 113461531A CN 202110668807 A CN202110668807 A CN 202110668807A CN 113461531 A CN113461531 A CN 113461531A
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- ultrafiltration
- rosmarinic acid
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- DOUMFZQKYFQNTF-WUTVXBCWSA-N (R)-rosmarinic acid Chemical compound C([C@H](C(=O)O)OC(=O)\C=C\C=1C=C(O)C(O)=CC=1)C1=CC=C(O)C(O)=C1 DOUMFZQKYFQNTF-WUTVXBCWSA-N 0.000 title claims abstract description 449
- DOUMFZQKYFQNTF-MRXNPFEDSA-N rosemarinic acid Natural products C([C@H](C(=O)O)OC(=O)C=CC=1C=C(O)C(O)=CC=1)C1=CC=C(O)C(O)=C1 DOUMFZQKYFQNTF-MRXNPFEDSA-N 0.000 title claims abstract description 212
- ZZAFFYPNLYCDEP-HNNXBMFYSA-N Rosmarinsaeure Natural products OC(=O)[C@H](Cc1cccc(O)c1O)OC(=O)C=Cc2ccc(O)c(O)c2 ZZAFFYPNLYCDEP-HNNXBMFYSA-N 0.000 title claims abstract description 211
- TVHVQJFBWRLYOD-UHFFFAOYSA-N rosmarinic acid Natural products OC(=O)C(Cc1ccc(O)c(O)c1)OC(=Cc2ccc(O)c(O)c2)C=O TVHVQJFBWRLYOD-UHFFFAOYSA-N 0.000 title claims abstract description 211
- 238000000034 method Methods 0.000 title claims abstract description 48
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 263
- 239000000243 solution Substances 0.000 claims abstract description 115
- 239000007788 liquid Substances 0.000 claims abstract description 72
- 239000011347 resin Substances 0.000 claims abstract description 55
- 229920005989 resin Polymers 0.000 claims abstract description 55
- 239000011575 calcium Substances 0.000 claims abstract description 41
- 239000003480 eluent Substances 0.000 claims abstract description 41
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims abstract description 35
- 229910052791 calcium Inorganic materials 0.000 claims abstract description 35
- 238000000926 separation method Methods 0.000 claims abstract description 30
- 239000000843 powder Substances 0.000 claims abstract description 29
- 239000002699 waste material Substances 0.000 claims abstract description 24
- 241001529742 Rosmarinus Species 0.000 claims abstract description 20
- 239000011550 stock solution Substances 0.000 claims abstract description 17
- 239000003963 antioxidant agent Substances 0.000 claims abstract description 15
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 128
- 239000000706 filtrate Substances 0.000 claims description 104
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 67
- 238000005086 pumping Methods 0.000 claims description 65
- 239000008213 purified water Substances 0.000 claims description 65
- 235000019441 ethanol Nutrition 0.000 claims description 41
- 239000012528 membrane Substances 0.000 claims description 39
- 238000002360 preparation method Methods 0.000 claims description 39
- 238000005406 washing Methods 0.000 claims description 37
- 239000012465 retentate Substances 0.000 claims description 35
- 239000012530 fluid Substances 0.000 claims description 33
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 claims description 25
- 238000001914 filtration Methods 0.000 claims description 25
- 238000003756 stirring Methods 0.000 claims description 20
- 238000007599 discharging Methods 0.000 claims description 16
- 239000002245 particle Substances 0.000 claims description 16
- 239000000725 suspension Substances 0.000 claims description 15
- 239000012141 concentrate Substances 0.000 claims description 13
- 238000010828 elution Methods 0.000 claims description 11
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 11
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- 239000003513 alkali Substances 0.000 claims description 10
- 238000010790 dilution Methods 0.000 claims description 10
- 239000012895 dilution Substances 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 9
- 239000002244 precipitate Substances 0.000 claims description 9
- 238000001556 precipitation Methods 0.000 claims description 9
- 102000004169 proteins and genes Human genes 0.000 claims description 7
- 108090000623 proteins and genes Proteins 0.000 claims description 7
- 238000001291 vacuum drying Methods 0.000 claims description 7
- 150000004676 glycans Chemical class 0.000 claims description 6
- 229920001282 polysaccharide Polymers 0.000 claims description 6
- 239000005017 polysaccharide Substances 0.000 claims description 6
- 239000000920 calcium hydroxide Substances 0.000 claims description 5
- 229910001861 calcium hydroxide Inorganic materials 0.000 claims description 5
- 239000011248 coating agent Substances 0.000 claims description 5
- 238000000576 coating method Methods 0.000 claims description 5
- 150000001875 compounds Chemical class 0.000 claims description 5
- 239000003337 fertilizer Substances 0.000 claims description 5
- 238000011049 filling Methods 0.000 claims description 5
- 238000007667 floating Methods 0.000 claims description 5
- 238000006386 neutralization reaction Methods 0.000 claims description 5
- 210000002966 serum Anatomy 0.000 claims description 5
- 238000002791 soaking Methods 0.000 claims description 5
- 239000002253 acid Substances 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 3
- 238000000605 extraction Methods 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 239000002994 raw material Substances 0.000 abstract description 4
- 239000003814 drug Substances 0.000 abstract description 3
- 238000003912 environmental pollution Methods 0.000 abstract description 3
- 235000013305 food Nutrition 0.000 abstract description 3
- 241000196324 Embryophyta Species 0.000 abstract description 2
- 238000005516 engineering process Methods 0.000 abstract description 2
- 238000002390 rotary evaporation Methods 0.000 description 15
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 11
- 239000012535 impurity Substances 0.000 description 9
- 239000000047 product Substances 0.000 description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- QRYRORQUOLYVBU-VBKZILBWSA-N carnosic acid Chemical compound CC([C@@H]1CC2)(C)CCC[C@]1(C(O)=O)C1=C2C=C(C(C)C)C(O)=C1O QRYRORQUOLYVBU-VBKZILBWSA-N 0.000 description 4
- 239000013049 sediment Substances 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 239000011148 porous material Substances 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- 244000178231 Rosmarinus officinalis Species 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 238000013375 chromatographic separation Methods 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 239000002360 explosive Substances 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 150000002772 monosaccharides Chemical class 0.000 description 2
- 239000003208 petroleum Substances 0.000 description 2
- -1 phenolic acid compound Chemical class 0.000 description 2
- YQUVCSBJEUQKSH-UHFFFAOYSA-N protochatechuic acid Natural products OC(=O)C1=CC=C(O)C(O)=C1 YQUVCSBJEUQKSH-UHFFFAOYSA-N 0.000 description 2
- WKOLLVMJNQIZCI-UHFFFAOYSA-N vanillic acid Chemical compound COC1=CC(C(O)=O)=CC=C1O WKOLLVMJNQIZCI-UHFFFAOYSA-N 0.000 description 2
- TUUBOHWZSQXCSW-UHFFFAOYSA-N vanillic acid Natural products COC1=CC(O)=CC(C(O)=O)=C1 TUUBOHWZSQXCSW-UHFFFAOYSA-N 0.000 description 2
- WCGUUGGRBIKTOS-GPOJBZKASA-N (3beta)-3-hydroxyurs-12-en-28-oic acid Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CC[C@@H](C)[C@H](C)[C@H]5C4=CC[C@@H]3[C@]21C WCGUUGGRBIKTOS-GPOJBZKASA-N 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 101900297506 Human immunodeficiency virus type 1 group M subtype B Reverse transcriptase/ribonuclease H Proteins 0.000 description 1
- 241000207923 Lamiaceae Species 0.000 description 1
- 241000191938 Micrococcus luteus Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 238000003889 chemical engineering Methods 0.000 description 1
- 238000002485 combustion reaction Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000004880 explosion Methods 0.000 description 1
- 230000007760 free radical scavenging Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 229910017053 inorganic salt Inorganic materials 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 239000002808 molecular sieve Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 1
- 229940096998 ursolic acid Drugs 0.000 description 1
- PLSAJKYPRJGMHO-UHFFFAOYSA-N ursolic acid Natural products CC1CCC2(CCC3(C)C(C=CC4C5(C)CCC(O)C(C)(C)C5CCC34C)C2C1C)C(=O)O PLSAJKYPRJGMHO-UHFFFAOYSA-N 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/24—Antidepressants
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
- A61P31/18—Antivirals for RNA viruses for HIV
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
- B01D61/14—Ultrafiltration; Microfiltration
- B01D61/145—Ultrafiltration
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D65/00—Accessories or auxiliary operations, in general, for separation processes or apparatus using semi-permeable membranes
- B01D65/02—Membrane cleaning or sterilisation ; Membrane regeneration
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- C07C67/52—Separation; Purification; Stabilisation; Use of additives by change in the physical state, e.g. crystallisation
- C07C67/54—Separation; Purification; Stabilisation; Use of additives by change in the physical state, e.g. crystallisation by distillation
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- C07C67/60—Separation; Purification; Stabilisation; Use of additives by treatment giving rise to chemical modification
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Abstract
A method for extracting rosmarinic acid belongs to the technical field of plant extraction. The method takes waste liquid generated by extracting rosemary fat-soluble antioxidant from dry rosemary leaves as a raw material, prepares rosmarinic acid stock solution, prepares rosmarinic acid ultrafiltration clear solution, prepares rosmarinic acid ultrafiltration concentrated solution, prepares rosmarinic acid calcium, prepares rosmarinic acid dissolved solution, prepares rosmarinic acid eluent and prepares rosmarinic acid dry powder, and prepares the rosemary dry powder with the mass content percentage of 89-98% and the total yield of 85-93%. The rosmarinic acid dry powder obtained by the invention has high rosmarinic acid content, good separation effect and high yield; the MCI resin used in the invention can be reused after being processed, has no environmental pollution and is a typical green production technology; the invention uses standard equipment which accords with GMP standard, has the advantages of standard operation, mild condition and low production cost, and the prepared rosmarinic acid powder product can be widely applied to the fields of food, health care products, medicine and the like.
Description
One, the technical field
The invention belongs to the technical field of plant extraction, and particularly relates to a method for preparing rosmarinic acid.
Second, background Art
Rosmarinic acid is a multifunctional phenolic acid compound existing in various labiatae plants, has strong in vivo free radical scavenging activity and antioxidation, has remarkable activities of inhibiting reproduction of bacillus subtilis, micrococcus luteus and escherichia coli, inhibiting HIV-1 reverse transcriptase, resisting depression and the like, has important application value in the fields of food, medicine, cosmetics and the like, and is widely concerned.
The existing method for preparing rosmarinic acid from rosemary, for example, a paper written by Zhang Lin and the like published in 2016, 19 th of Guangzhou chemical engineering, introduces a method for separating and purifying rosmarinic acid from dry rosemary leaves, namely, dry rosemary leaves are taken as raw materials, extracted for 2 times by 50% ethanol, then part of impurities are removed by microfiltration, then n-butanol is used for extraction, then ethyl acetate is used for extraction, extract liquor is combined, solvent is recovered under reduced pressure, dry powder is added with water for dissolution, then X-5 macroporous resin is used for separation and enrichment, finally recrystallization is carried out to obtain a product, and the purity reaches 66.67%. The disadvantages of this method are: firstly, only rosmarinic acid is extracted, which causes huge resource waste of fat-soluble antioxidants, namely carnosic acid and ursolic acid, which are main components in dry rosemary leaves; the steps are complicated, and the energy consumption is high; ethyl acetate has low boiling point and large peculiar smell, and is quick to volatilize at normal temperature, flammable and explosive, high in danger and serious in environmental pollution; fourthly, the extraction rate of the ethyl acetate and the n-butyl alcohol to the rosmarinic acid is low, and the single extraction rate is only 22 percent and 41 percent respectively. In addition, the boiling point of the n-butyl alcohol is as high as 110 ℃, and the n-butyl alcohol is easy to remain in the rosmarinic acid to cause product pollution. For another example, patent 201010130287.1 states that during the chromatographic separation, petroleum ether 2-3 times the column volume, ethyl acetate 2-8 times the column volume, 30% ethanol 2-5 times the column volume, and 80% ethanol 2-3 times the column volume are sequentially used for elution. The disadvantages of this method are: the method has the advantages of various steps and complex operation; consumption of organic solvent increases cost and pollutes environment; petroleum ether has strong irritation and high flammability, can form explosive mixture with air, and can cause combustion and explosion when meeting open fire and high heat. Not only can threaten the health of operators, but also has larger potential safety hazard.
Third, the invention
Aiming at the defects of the existing method for preparing the rosmarinic acid, the invention provides a method for preparing the rosmarinic acid by taking waste liquid generated in the process of extracting the rosemary fat-soluble antioxidant as a raw material. The method has the characteristics of simple operation, high efficiency, environmental protection, high product yield, high rosmarinic acid content and the like.
The main principle of the invention is as follows: the carboxyl group of rosmarinic acid has negative charge and can react with Ca2+Etc. form a precipitate which is insoluble or poorly soluble in water, and therefore, Ca-containing is added to an aqueous solution containing rosmarinic acid2+And the like, and calcium rosmarinate precipitates can be generated and separated from the solution. The membrane separation is a precise separation technology, and uses the pressure difference between two sides of the membrane as driving force, and utilizes the selective permeability of membrane pore size to make the solute whose molecular diameter is less than its pore size permeate through it, and can retain the solute whose molecular diameter is greater than its pore size so as to implement separation and classification. Rosemary (Rosemary)The molecular weight of the vanillic acid is 360.13Da, the vanillic acid is far less than saccharides with the molecular weight of more than 50kDa, proteins with the molecular weight of more than 10kDa, and inorganic salts, monosaccharides and other small molecular impurities with the molecular weight of more than 200 Da. Therefore, impurity molecules which are obviously larger or obviously smaller than the rosmarinic acid can be separated through membrane separation, and the content of the rosmarinic acid in the separated matter is improved. The MCI resin is also called medium pressure chromatographic separation gel, has a porous structure, and the separation principle is as follows: when the molecular diameter of the solute is larger than that of the resin micropores, the solute cannot enter the interior of the resin micropores and flows out of the column along with the solvent from gaps among the resins; molecules with solute molecule diameter smaller than that of the micropores can freely enter and exit the particles; the diameter of solute molecule is close to the diameter of MCI resin micropore, and only superficial surface is reached, and then the solute molecule is washed away by solvent, and the process is repeated. Therefore, solutes having significantly different molecular diameters have significantly different paths and distances through the MCI resin column, and thus separation of the target from impurities occurs.
The molecular weight of the rosmarinic acid is only 360.13Da, the diameter of the rosmarinic acid is far smaller than that of macromolecular impurities such as protein and polysaccharide with the molecular weight of tens of thousands Da, and is also obviously larger than that of micromolecular impurities such as amino acid, monosaccharide, inorganic salt and the like, and the paths of the rosmarinic acid in the MCI resin column are obviously different, so that the eluent with the characteristic absorption of the rosmarinic acid is collected, and the high-purity rosmarinic acid product can be prepared.
The purpose of the invention is realized as follows: a method for preparing rosmarinic acid comprises the steps of taking waste liquid generated by extracting a rosemary fat-soluble antioxidant from dry leaves of rosemary as a raw material, and preparing rosmarinic acid stock solution by (1), preparing rosmarinic acid ultrafiltration clear liquid by (2), preparing rosmarinic acid ultrafiltration concentrated solution by (3), preparing rosmarinic acid calcium by (4), preparing rosmarinic acid dissolving solution by (5), preparing rosmarinic acid eluent by (6), and preparing rosmarinic acid dry powder by (7), so that the rosmarinic acid dry powder with the purity of 89-98% and the total yield of 85-93% is prepared. The specific process steps are as follows:
(1) preparation of a Rosmarinic acid stock solution
The centrifuged clear solution obtained in step (2) is prepared according to the method introduced in the Chinese patent publication No. CN 107739305A, example 1, and is the waste liquid produced in the process of producing rosemary lipid-soluble antioxidant. And uniformly mixing the waste liquid with purified water, wherein the volume ratio of the waste liquid to the purified water is 1L: 5-8L. Adjusting the pH value to 3-5 by using dilute hydrochloric acid, pumping the mixture into a tubular centrifuge, separating at the speed of 15000-20000 r/min, respectively collecting centrifugal clear liquid and centrifugal precipitate, and preparing the collected centrifugal clear liquid, namely rosmarinic acid stock solution, for the next step to prepare rosmarinic acid ultrafiltration clear liquid; and (4) carrying out centrifugal precipitation on the collected sediment for preparing the natural mildew-proof coating.
(2) Preparation of Rosmarinic acid Ultrafiltration serum
And (3) after the step (1) is finished, pumping the rosmarinic acid stock solution prepared in the step (1) into an ultrafilter with the molecular weight cutoff of 10-50 KDa, and carrying out primary ultrafiltration under the condition that the gauge pressure is 0.2-0.6 MPa until the volume ratio of the primary ultrafiltration cutoff solution to the filtrate is 1L: 3-5L. Collecting the first ultrafiltration retentate and the first ultrafiltration filtrate, mixing the collected first ultrafiltration filtrate with the filtrate obtained by the subsequent second ultrafiltration, and using the filtrate to prepare rosmarinic acid ultrafiltration concentrate; adding purified water into the collected first ultrafiltration trapped fluid, carrying out second ultrafiltration, wherein the volume ratio of the first ultrafiltration trapped fluid to the added purified water is 1L: 3-5L, the second ultrafiltration gauge pressure is 0.2-0.6 MPa, stopping ultrafiltration until the volume ratio of the second ultrafiltration trapped fluid to the filtrate thereof is 1L: 3-5L, respectively collecting the second ultrafiltration filtrate, the second ultrafiltration trapped fluid and the ultrafiltration membrane with the molecular weight cutoff after the second ultrafiltration separation of 10-50 KDa, washing for 15-20 min with purified water, then washing for 15-20 min with 0.5-1% alkali liquor, and finally washing with purified water until the pH value is 5-7, thus obtaining a regenerated ultrafiltration membrane which can be used for preparing next batch of rosmarinic acid ultrafiltration clear liquid.
And collecting the second ultrafiltration filtrate, and combining the second ultrafiltration filtrate with the collected first ultrafiltration filtrate to obtain rosmarinic acid ultrafiltration clear liquid for preparing the rosmarinic acid ultrafiltration concentrated solution in the next step. And pumping the collected second ultrafiltration trapped fluid mainly containing macromolecular polysaccharide, protein and the like into a biochemical tank for biochemical treatment, and discharging after reaching the standard.
(3) Preparation of Rosmarinic acid ultrafiltered concentrate
And (3) after the step (2) is finished, pumping the rosmarinic acid ultrafiltration clear liquid prepared in the step (2) into an ultrafilter with molecular weight cutoff of 500-1000 Da, and carrying out primary ultrafiltration under the condition that the gauge pressure is 0.2-0.6 MPa until the volume ratio of the primary ultrafiltration cut-off liquid to the primary ultrafiltration filtrate is 1L: 3-5L. Respectively collecting the first ultrafiltration trapped fluid and the first ultrafiltration filtered fluid, pumping the collected first ultrafiltration filtered fluid into a biochemical tank for biochemical treatment, and discharging after reaching the standard; and adding purified water into the collected first ultrafiltration trapped fluid for dilution, wherein the volume ratio of the first ultrafiltration trapped fluid to the added purified water is 1L: 3-5L. And after the dilution is finished, carrying out second ultrafiltration until the volume ratio of the second ultrafiltration retentate to the second ultrafiltration filtrate is 1L: 3-5L, and stopping ultrafiltration. Respectively collecting the second ultrafiltration filtrate and the second ultrafiltration retentate, and collecting the second ultrafiltration retentate to obtain the rosmarinic acid ultrafiltration concentrate for preparing calcium rosmarinic acid; pumping the collected second ultrafiltration filtrate into a biochemical treatment tank for biochemical treatment, and discharging after reaching the standard. And (3) washing the collected ultrafiltration membrane with the molecular weight cutoff of 500-1000 Da after the second ultrafiltration separation for 15-20 min by using purified water, then washing the ultrafiltration membrane for 15-20 min by using 0.5-1% alkali liquor, and finally washing the ultrafiltration membrane by using purified water until the pH value is 5-7, so as to obtain a regenerated ultrafiltration membrane, wherein the membrane can be used for preparing the next batch of rosmarinic acid ultrafiltration concentrated solution.
(4) Preparation of calcium Rosmarinate
Dissolving appropriate amount of calcium hydroxide in purified water, filtering, and collecting filtrate to obtain dilute calcium hydroxide solution containing Ca2+The mass percentage concentration is 0.1-0.5%, and the pH is 8-10. After the step (3) is finished, pumping the rosmarinic acid ultrafiltration concentrated solution prepared in the step (3) into a neutralization tank, controlling the stirring speed to be 50-100 r/min, and dropwise adding Ca2+And (3) a dilute calcium hydroxide solution with the mass percentage concentration of 0.1-0.5% and the pH of 8-10, stopping dropwise adding the dilute calcium hydroxide solution when obvious precipitation appears in the ultrafiltration concentrated solution and the pH value reaches 8-10, and thus preparing the rosmarinic acid calcium suspension solution. Ca was added dropwise2+The mass percentage concentration of the dilute calcium hydroxide solution is 0.1-0.5%, and the pH value is 8-10Then, the stirring is stopped and the suspension is kept still for 6 to 12 hours. Finally, filtering the suspension solution, respectively collecting filter residues and filtrate, and performing biochemical treatment on the collected filtrate to discharge after reaching the standard; and preparing the calcium rosmarinate from the collected filter residues for preparing a rosmarinic acid dissolving solution in the next step.
(5) Preparation of a solution of rosmarinic acid
And (4) after the step (4) is finished, dispersing the calcium rosmarinate prepared in the step (4) into purified water, wherein the mass ratio of the calcium rosmarinate to the volume ratio of the purified water is 1 g: 500-1000 mL, adding the dilute hydrochloric acid solution used in the step (1) for preparing the raw rosmarinate solution under the condition that the stirring speed of a dissolving tank is 50-100 r/min, and stopping dripping and stirring when the pH value of the system is 4-6. Then, filtration was performed until no filtrate flowed out. Collecting filtrate and filtering residue, and collecting filtrate to obtain rosmarinic acid solution for further preparation of rosmarinic acid eluate; and (4) using the collected filter residues for preparing the compound fertilizer.
(6) Preparation of Rosmarinic acid eluent
The method comprises the steps of taking MCI GEL CHP 20 type resin with the particle size specification of 35-75 mu m, soaking the resin in absolute ethyl alcohol with the volume (BV) being 4-6 times of that of the resin for 4-8 hours, removing resin particles floating on the upper layer, filtering, and collecting filter residues and filtrate respectively. Recovering ethanol from the collected filtrate; and balancing the collected filter residues for 1-2 hours by using an ethanol water solution with 4-6 times volume (BV), pH of 3-5 and volume percentage concentration of 40-60%, then filling the filter residues into a column by a wet method, and connecting the column with a constant flow pump, an ultraviolet detector and an automatic fraction collector to form an MCI resin column separation system for later use.
And (5) after the step (5) is finished, pumping the rosmarinic acid dissolving solution prepared in the step (5) into an activated MCI GEL CHP 20 type resin column separation system with the particle size of 35-75 mu m, controlling the pumping flow rate of the rosmarinic acid dissolving solution to be 2-5 times of the resin volume (BV)/h, and controlling the ratio of the MCI resin volume to the dissolving solution volume to be 1L: 10-15L. And after the rosmarinic acid dissolving solution is pumped, pumping an ethanol water solution with the ethanol volume fraction of 70-90% for elution, wherein the pumping flow rate is 1-5 BV/h, and respectively collecting eluents of 1-4 BV, 5-8 BV and 9-12 BV. Combining the collected eluates of 1BV to 4BV and 9BV to 12BV for recovering ethanol; and (3) collecting the eluent of the 5 th BV to 8 th BV, namely the rosmarinic acid eluent, and using the eluent to prepare rosmarinic acid powder in the next step. And after the elution is finished, pumping an ethanol water solution with the ethanol volume fraction of 90-95% for washing, wherein the pumping volume of the ethanol water solution is 1-6 BV, and the pumping flow rate is 1-5 BV/h. The washed MCI resin column can be used for preparing rosmarinic acid eluent in the next batch.
(7) Preparation of Rosmarinic acid powder
And (6) after the step (6) is finished, pumping the rosmarinic acid eluent prepared in the step (6) into a rotary evaporator for reduced pressure concentration, distilling ethanol at the temperature of 60-70 ℃ and the relative vacuum degree of-0.06-0.09 MPa, stopping reduced pressure concentration when no condensed liquid flows out of a condenser, and respectively collecting a condensate and a concentrated solution. The collected condensate is used for recovering ethanol. And transferring the collected concentrated solution into a vacuum drier, and carrying out vacuum drying for 5-8 h under the conditions that the temperature is 45-50 ℃ and the relative vacuum degree is-0.04-0.08 MPa, thus preparing the rosmarinic acid powder. Wherein the mass percentage of the rosmarinic acid is 89-98%, and the total yield of the rosmarinic acid powder is 85-93% of the mass of the rosmarinic acid in the waste liquid generated in the process of producing the rosemary lipid-soluble antioxidant.
After the technical scheme is adopted, the invention mainly has the following effects:
1. good separation effect and high yield. The invention uses ultrafiltration membrane to separate macromolecular impurities and micromolecular impurities in the rosmarinic acid extracting solution, and forms calcium rosmarinic acid to further separate impurities with similar properties; and finally purifying by using MCI resin according to the principle of a molecular sieve to obtain powder with the mass percentage of the rosmarinic acid up to 89-98%, wherein the purification multiple is 11.13-12.25. The separation efficiency is high, the effect is good, the total yield of the rosmarinic acid is as high as 85-93%, and the yield is high.
2. Realizes comprehensive utilization and is green and environment-friendly. In the implementation process of the invention, the byproducts such as centrifugal precipitate and the like generated in the process of producing the rosemary lipid-soluble antioxidant carnosic acid are respectively subjected to deep processing treatment, so that the comprehensive utilization is realized; the used solvent is only ethanol, so the paint is non-toxic and has low peculiar smell; the used MCI resin can be reused after being treated; the generated small amount of waste water is discharged after reaching the standard after biochemical treatment, and basically has no environmental pollution.
3. The production equipment is standardized, the cost is low, and the product has wide application. The invention uses standard equipment which accords with GMP standard, has standard operation, mild condition and low production cost, and the prepared rosmarinic acid powder can be widely applied to the fields of food, health care products, medicine and the like and has huge social and economic values.
Fourth, detailed description of the invention
The present invention will be further described with reference to the following specific embodiments.
Example 1
A method for extracting rosmarinic acid comprises the following specific process steps:
(1) preparation of a Rosmarinic acid stock solution
The centrifuged clear solution obtained in step (2) is prepared according to the method introduced in the Chinese patent publication No. CN 107739305A, example 1, and is the waste liquid produced in the process of producing rosemary lipid-soluble antioxidant. The waste liquid and purified water are uniformly mixed, and the volume ratio of the waste liquid to the purified water is 1L: 8L. Adjusting pH to 4 with dilute hydrochloric acid, pumping into tubular centrifuge, separating at 20000r/min, collecting centrifugated clear liquid and centrifugated precipitate respectively, and collecting centrifugated clear liquid to obtain rosmarinic acid stock solution for use in the next step to prepare rosmarinic acid ultrafiltered clear liquid; and (4) carrying out centrifugal precipitation on the collected sediment for preparing the natural mildew-proof coating.
(2) Preparation of Rosmarinic acid Ultrafiltration serum
And (3) after the step (1) is finished, pumping the rosmarinic acid stock solution prepared in the step (1) into an ultrafilter with the molecular weight cutoff of 50KDa, and carrying out primary ultrafiltration under the condition that the gauge pressure is 0.6MPa until the volume ratio of the primary ultrafiltration cutoff solution to the filtrate is 1L: 5L. Collecting the first ultrafiltration retentate and the first ultrafiltration filtrate, mixing the collected first ultrafiltration filtrate with the filtrate obtained by the subsequent second ultrafiltration, and using the filtrate to prepare rosmarinic acid ultrafiltration concentrate; adding purified water into the collected first ultrafiltration retentate, performing second ultrafiltration, wherein the volume ratio of the first ultrafiltration retentate to the added purified water is 1L: 5L, the second ultrafiltration gauge pressure is 0.6MPa, stopping ultrafiltration until the volume ratio of the second ultrafiltration retentate to the filtrate is 1L: 5L, respectively collecting the second ultrafiltration filtrate, the second ultrafiltration retentate and ultrafiltration membrane with the cut-off molecular weight of 50KDa after the second ultrafiltration separation, washing with purified water for 120min, then washing with 0.5% alkali liquor for 20min, and finally washing with purified water until the pH value is 7, thus obtaining a regenerated ultrafiltration membrane, which can be used for preparing next batch of rosmarinic acid ultrafiltration clear liquid.
And collecting the second ultrafiltration filtrate, and combining the second ultrafiltration filtrate with the collected first ultrafiltration filtrate to obtain rosmarinic acid ultrafiltration clear liquid for preparing the rosmarinic acid ultrafiltration concentrated solution in the next step. And pumping the collected second ultrafiltration trapped fluid mainly containing macromolecular polysaccharide, protein and the like into a biochemical tank for biochemical treatment, and discharging after reaching the standard.
(3) Preparation of Rosmarinic acid ultrafiltered concentrate
And (3) after the step (2) is finished, pumping the rosmarinic acid ultrafiltration clear liquid prepared in the step (2) into an ultrafilter with the molecular weight cutoff of 1000Da, and carrying out primary ultrafiltration under the condition that the gauge pressure is 0.6MPa until the volume ratio of the primary ultrafiltration cut-off liquid to the primary ultrafiltration filtrate is 1L: 5L. Respectively collecting the first ultrafiltration trapped fluid and the first ultrafiltration filtered fluid, pumping the collected first ultrafiltration filtered fluid into a biochemical tank for biochemical treatment, and discharging after reaching the standard; and adding purified water into the collected first ultrafiltration retentate for dilution, wherein the volume ratio of the first ultrafiltration retentate to the added purified water is 1L: 5L. And after the dilution is finished, carrying out second ultrafiltration until the volume ratio of the second ultrafiltration retentate to the second ultrafiltration filtrate is 1L: 5L, and stopping ultrafiltration. Respectively collecting the second ultrafiltration filtrate and the second ultrafiltration retentate, and collecting the second ultrafiltration retentate to obtain the rosmarinic acid ultrafiltration concentrate for preparing calcium rosmarinic acid; pumping the collected second ultrafiltration filtrate into a biochemical treatment tank for biochemical treatment, and discharging after reaching the standard. And (3) washing the collected ultrafiltration membranes with the molecular weight cut-off of 1000Da respectively after the second ultrafiltration separation for 20min by using purified water, then washing the ultrafiltration membranes for 20min by using 0.5% alkali liquor, and finally washing the ultrafiltration membranes to the pH value of 7 by using the purified water to obtain regenerated ultrafiltration membranes, wherein the membranes can be used for preparing the next batch of rosmarinic acid ultrafiltration concentrated solution.
(4) Preparation of calcium Rosmarinate
Dissolving appropriate amount of calcium hydroxide in purified water, filtering, and collecting filtrate to obtain dilute calcium hydroxide solution containing Ca2+The mass percentage concentration is 0.1 percent, and the pH value is 8. After the step (3) is finished, pumping the rosmarinic acid ultrafiltration concentrated solution prepared in the step (3) into a neutralization tank, controlling the stirring speed at 100r/min, and dropwise adding Ca2+And (3) diluting the calcium hydroxide solution with the mass percentage concentration of 0.1% and the pH value of 8 until obvious precipitation appears in the ultrafiltration concentrated solution, and stopping dropwise adding the diluted calcium hydroxide solution when the pH value reaches 8 to prepare the calcium rosmarinate suspension solution. Ca was added dropwise2+After completion of the dilute calcium hydroxide solution having a mass percentage concentration of 0.1% and a pH of 8, the stirring was stopped and the suspension was allowed to stand for 12 hours. Finally, filtering the suspension solution, respectively collecting filter residues and filtrate, and performing biochemical treatment on the collected filtrate to discharge after reaching the standard; and preparing the calcium rosmarinate from the collected filter residues for preparing a rosmarinic acid dissolving solution in the next step.
(5) Preparation of a solution of rosmarinic acid
And (4) after the step (4) is finished, dispersing the calcium rosmarinate prepared in the step (4) into purified water, wherein the mass ratio of the calcium rosmarinate to the volume of the purified water is 1 g: 1000mL, adding the dilute hydrochloric acid solution used in the step (1) for preparing the raw rosmarinate solution under the condition that the stirring speed of a dissolving tank is 100r/min, and stopping dripping and stirring when the pH value of the system is 4. Then, filtration was performed until no filtrate flowed out. Collecting filtrate and filtering residue, and collecting filtrate to obtain rosmarinic acid solution for further preparation of rosmarinic acid eluate; and (4) using the collected filter residues for preparing the compound fertilizer.
(6) Preparation of Rosmarinic acid eluent
Soaking MCI GEL CHP 20 type resin with particle size of 35 μm in anhydrous ethanol 4 times the volume of resin (BV) for 8 hr, removing floating resin particles on the upper layer, filtering, and collecting residue and filtrate. Recovering ethanol from the collected filtrate; and balancing the collected filter residue with 4 times volume (BV) of ethanol aqueous solution with pH of 3 and volume percentage concentration of 40% for 1 hour, filling the filter residue into a column by a wet method, and connecting the column with a constant flow pump, an ultraviolet detector and an automatic fraction collector to form an MCI resin column separation system for later use.
After the step (5) is finished, the rosmarinic acid dissolving solution prepared in the step (5) is pumped into a resin column separation system filled with activated MCI GEL CHP 20 type resin with the particle size of 35 mu m, the pumping flow rate of the rosmarinic acid dissolving solution is controlled to be 2 times of the resin volume (BV)/h, and the MCI resin volume and the dissolving solution volume ratio is 1L: 10L. And after the rosmarinic acid solution is pumped, pumping an ethanol water solution with the ethanol volume fraction of 70% for elution, wherein the pumping flow rate is 3BV/h, and respectively collecting eluates of the 1BV, the 5BV and the 9 BV. Combining the collected 1BV and 9BV eluents for recovering ethanol; and collecting the eluent of the 5BV, namely the rosmarinic acid eluent, and using the eluent to prepare rosmarinic acid powder in the next step. After the elution was completed, washing was carried out by pumping in an aqueous ethanol solution having an ethanol volume fraction of 95%, the volume of which was 2BV, and the flow rate was 1 BV/h. After the washing is completed, the MCI resin column can be used for the next batch to prepare the rosmarinic acid eluent.
(7) Preparation of Rosmarinic acid powder
And (6) after the step (6) is finished, pumping the rosmarinic acid eluent prepared in the step (6) into a rotary evaporator for reduced pressure concentration, distilling ethanol at the temperature of 70 ℃ and the relative vacuum degree of-0.09 MPa, stopping rotary evaporation when no condensed liquid flows out of a condenser, and respectively collecting rotary evaporation condensate and rotary evaporation concentrated liquid. And the rotary evaporation condensate can be recycled to an ethanol storage tank. And transferring the collected rotary evaporation concentrated solution into a vacuum drier for vacuum drying, and performing vacuum drying for 8 hours at the temperature of 45 ℃ under the relative vacuum degree of-0.08 MPa to prepare rosmarinic acid powder. Wherein the content of rosmarinic acid is 98%, and the total yield of the rosmarinic acid powder is 85% of the mass of rosmarinic acid in the waste liquid generated in the process of producing the rosemary fat-soluble antioxidant.
Example 2
A method for extracting rosmarinic acid comprises the following specific process steps:
(1) preparation of a Rosmarinic acid stock solution
The centrifuged clear solution obtained in step (2) is prepared according to the method introduced in the Chinese patent publication No. CN 107739305A, example 1, and is the waste liquid produced in the process of producing rosemary lipid-soluble antioxidant. Mixing the waste liquid and purified water uniformly, wherein the volume ratio of the waste liquid to the purified water is 1L: 5L. Adjusting pH to 4 with dilute hydrochloric acid, pumping into tubular centrifuge, separating at 20000r/min, collecting centrifugated clear liquid and centrifugated precipitate respectively, and collecting centrifugated clear liquid to obtain rosmarinic acid stock solution for use in the next step to prepare rosmarinic acid ultrafiltered clear liquid; and (4) carrying out centrifugal precipitation on the collected sediment for preparing the natural mildew-proof coating.
(2) Preparation of Rosmarinic acid Ultrafiltration serum
And (3) after the step (1) is finished, pumping the rosmarinic acid stock solution prepared in the step (1) into an ultrafilter with the molecular weight cutoff of 10KDa, and carrying out primary ultrafiltration under the condition that the gauge pressure is 0.6MPa until the volume ratio of the primary ultrafiltration cutoff solution to the filtrate is 1L: 4L. Collecting the first ultrafiltration retentate and the first ultrafiltration filtrate, mixing the collected first ultrafiltration filtrate with the filtrate obtained by the subsequent second ultrafiltration, and using the filtrate to prepare rosmarinic acid ultrafiltration concentrate; adding purified water into the collected first ultrafiltration retentate, performing second ultrafiltration, wherein the volume ratio of the first ultrafiltration retentate to the added purified water is 1L: 4L, the second ultrafiltration gauge pressure is 0.6MPa, stopping ultrafiltration until the volume ratio of the second ultrafiltration retentate to the filtrate is 1L: 4L, respectively collecting the second ultrafiltration filtrate, the second ultrafiltration retentate and an ultrafiltration membrane with the cut-off molecular weight of 10KDa after the second ultrafiltration separation, washing with purified water for 20min, then washing with 0.5% alkali liquor for 20min, and finally washing with purified water until the pH value is 7, thus obtaining a regenerated ultrafiltration membrane, which can be used for preparing the next batch of rosmarinic acid ultrafiltration clear liquid.
And collecting the second ultrafiltration filtrate, and combining the second ultrafiltration filtrate with the collected first ultrafiltration filtrate to obtain rosmarinic acid ultrafiltration clear liquid for preparing the rosmarinic acid ultrafiltration concentrated solution in the next step. And pumping the collected second ultrafiltration trapped fluid mainly containing macromolecular polysaccharide, protein and the like into a biochemical tank for biochemical treatment, and discharging after reaching the standard.
(3) Preparation of Rosmarinic acid ultrafiltered concentrate
And (3) after the step (2), pumping the rosmarinic acid ultrafiltration clear liquid prepared in the step (2) into an ultrafilter with molecular weight cutoff of 500Da, and carrying out primary ultrafiltration under the condition that the gauge pressure is 0.6MPa until the volume ratio of the primary ultrafiltration cut-off liquid to the primary ultrafiltration filtrate is 1L: 4L. Respectively collecting the first ultrafiltration trapped fluid and the first ultrafiltration filtered fluid, pumping the collected first ultrafiltration filtered fluid into a biochemical tank for biochemical treatment, and discharging after reaching the standard; and adding purified water into the collected first ultrafiltration retentate for dilution, wherein the volume ratio of the first ultrafiltration retentate to the added purified water is 1L: 4L. And after the dilution is finished, carrying out second ultrafiltration until the volume ratio of the second ultrafiltration retentate to the second ultrafiltration filtrate is 1L: 4L, and stopping ultrafiltration. Respectively collecting the second ultrafiltration filtrate and the second ultrafiltration retentate, and collecting the second ultrafiltration retentate to obtain the rosmarinic acid ultrafiltration concentrate for preparing calcium rosmarinic acid; pumping the collected second ultrafiltration filtrate into a biochemical treatment tank for biochemical treatment, and discharging after reaching the standard. And (3) washing the collected ultrafiltration membrane with the molecular weight cutoff of 500Da after the second ultrafiltration separation for 20min by using purified water, then washing the ultrafiltration membrane for 20min by using 0.5% alkali liquor, and finally washing the ultrafiltration membrane by using the purified water until the pH value is 7 to obtain a regenerated ultrafiltration membrane, wherein the membrane can be used for preparing the next batch of rosmarinic acid ultrafiltration concentrated solution.
(4) Preparation of calcium Rosmarinate
Dissolving appropriate amount of calcium hydroxide in purified water, filtering, and collecting filtrate to obtain dilute calcium hydroxide solution containing Ca2+The mass percentage concentration is 0.1 percent, and the pH value is 8. After the step (3) is finished, pumping the rosmarinic acid ultrafiltration concentrated solution prepared in the step (3) into a neutralization tank, controlling the stirring speed at 100r/min, and dropwise adding Ca2+The mass percentage concentration is 0.1 percentAnd the dilute calcium hydroxide solution with the pH value of 8 is added until obvious precipitation appears in the ultrafiltration concentrated solution, and the dilute calcium hydroxide solution is stopped to be added when the pH value reaches 8, so that the rosmarinic acid calcium suspension solution is prepared. Ca was added dropwise2+After completion of the dilute calcium hydroxide solution having a mass percentage concentration of 0.1% and a pH of 8, the stirring was stopped and the suspension was allowed to stand for 12 hours. Finally, filtering the suspension solution, respectively collecting filter residues and filtrate, and performing biochemical treatment on the collected filtrate to discharge after reaching the standard; and preparing the calcium rosmarinate from the collected filter residues for preparing a rosmarinic acid dissolving solution in the next step.
(5) Preparation of a solution of rosmarinic acid
And (4) after the step (4) is finished, dispersing the calcium rosmarinate prepared in the step (4) into purified water, wherein the mass ratio of the calcium rosmarinate to the volume of the purified water is 1 g: 800mL, adding the dilute hydrochloric acid solution used in the step (1) for preparing the raw rosmarinate solution under the condition that the stirring speed of a dissolving tank is 100r/min, and stopping dripping and stirring when the pH value of the system is 4. Then, filtration was performed until no filtrate flowed out. Collecting filtrate and filtering residue, and collecting filtrate to obtain rosmarinic acid solution for further preparation of rosmarinic acid eluate; and (4) using the collected filter residues for preparing the compound fertilizer.
(6) Preparation of Rosmarinic acid eluent
Soaking MCI GEL CHP 20 type resin with particle size of 55 μm in 5 times of resin volume (BV) of anhydrous ethanol for 6 hr, removing resin particles floating on the upper layer, filtering, and collecting residue and filtrate. Recovering ethanol from the collected filtrate; and balancing the collected filter residue with 5 times volume (BV) of ethanol aqueous solution with pH of 4 and volume percentage concentration of 60% for 1 hour, filling the filter residue into a column by a wet method, and connecting the column with a constant flow pump, an ultraviolet detector and an automatic fraction collector to form an MCI resin column separation system for later use.
After the step (5) is finished, the rosmarinic acid dissolving solution prepared in the step (5) is pumped into a resin column separation system filled with activated MCI GEL CHP 20 type resin with the particle size of 55 mu m, the pumping flow rate of the rosmarinic acid dissolving solution is controlled to be 4 times of the resin volume (BV)/h, and the MCI resin volume and the dissolving solution volume ratio is 1L: 10L. And after the rosmarinic acid solution is pumped, pumping an ethanol water solution with the ethanol volume fraction of 80% for elution, wherein the pumping flow rate is 2BV/h, and respectively collecting eluates of the 2BV, the 6BV and the 10 BV. Combining the collected eluent of the 2BV and the eluent of the 10BV for recovering the ethanol; and collecting the eluent of the 6BV, namely the rosmarinic acid eluent, and using the eluent to prepare rosmarinic acid powder in the next step. After the elution was completed, washing was carried out by pumping in an aqueous ethanol solution having an ethanol volume fraction of 95%, the volume of which was 2BV, and the flow rate was 1 BV/h. After the washing is completed, the MCI resin column can be used for the next batch to prepare the rosmarinic acid eluent.
(7) Preparation of Rosmarinic acid powder
And (6) after the step (6) is finished, pumping the rosmarinic acid eluent prepared in the step (6) into a rotary evaporator for reduced pressure concentration, distilling ethanol at the temperature of 70 ℃ and the relative vacuum degree of-0.09 MPa, stopping rotary evaporation when no condensed liquid flows out of a condenser, and respectively collecting rotary evaporation condensate and rotary evaporation concentrated liquid. And the rotary evaporation condensate can be recycled to an ethanol storage tank. And transferring the collected rotary evaporation concentrated solution into a vacuum drier for vacuum drying, and performing vacuum drying for 8 hours at the temperature of 45 ℃ under the relative vacuum degree of-0.08 MPa to prepare rosmarinic acid powder. Wherein the content of rosmarinic acid is 94%, and the total yield of the rosmarinic acid powder is 90% of the mass of rosmarinic acid in the waste liquid generated in the process of producing the rosemary lipid-soluble antioxidant.
Example 3
A method for extracting rosmarinic acid comprises the following specific process steps:
(1) preparation of a Rosmarinic acid stock solution
The centrifuged clear solution obtained in step (2) is prepared according to the method introduced in the Chinese patent publication No. CN 107739305A, example 1, and is the waste liquid produced in the process of producing rosemary lipid-soluble antioxidant. Mixing the waste liquid with purified water uniformly, wherein the volume ratio of the waste liquid to the purified water is 1L: 6L. Adjusting pH to 4 with dilute hydrochloric acid, pumping into tubular centrifuge, separating at 15000r/min, respectively collecting centrifugated clear liquid and centrifugated precipitate, and collecting centrifugated clear liquid to obtain rosmarinic acid stock solution for use in the next step to prepare rosmarinic acid ultrafiltered clear liquid; and (4) carrying out centrifugal precipitation on the collected sediment for preparing the natural mildew-proof coating.
(2) Preparation of Rosmarinic acid Ultrafiltration serum
And (3) after the step (1) is finished, pumping the rosmarinic acid stock solution prepared in the step (1) into an ultrafilter with the molecular weight cutoff of 30KDa, and carrying out primary ultrafiltration under the condition that the gauge pressure is 0.5MPa until the volume ratio of the primary ultrafiltration cutoff solution to the filtrate is 1L: 4L. Collecting the first ultrafiltration retentate and the first ultrafiltration filtrate, mixing the collected first ultrafiltration filtrate with the filtrate obtained by the subsequent second ultrafiltration, and using the filtrate to prepare rosmarinic acid ultrafiltration concentrate; adding purified water into the collected first ultrafiltration retentate, performing second ultrafiltration, wherein the volume ratio of the first ultrafiltration retentate to the added purified water is 1L: 4L, the second ultrafiltration gauge pressure is 0.6MPa, stopping ultrafiltration until the volume ratio of the second ultrafiltration retentate to the filtrate is 1L: 4L, respectively collecting the second ultrafiltration filtrate, the second ultrafiltration retentate and ultrafiltration membranes with the cut-off molecular weight of 30KDa after the second ultrafiltration separation, washing with purified water for 20min, then washing with 0.5% alkali liquor for 20min, and finally washing with purified water until the pH value is 6, thus obtaining regenerated ultrafiltration membranes which can be used for preparing next batch of rosmarinic acid ultrafiltration clear liquid.
And collecting the second ultrafiltration filtrate, and combining the second ultrafiltration filtrate with the collected first ultrafiltration filtrate to obtain rosmarinic acid ultrafiltration clear liquid for preparing the rosmarinic acid ultrafiltration concentrated solution in the next step. And pumping the collected second ultrafiltration trapped fluid mainly containing macromolecular polysaccharide, protein and the like into a biochemical tank for biochemical treatment, and discharging after reaching the standard.
(3) Preparation of Rosmarinic acid ultrafiltered concentrate
And (3) after the step (2), pumping the rosmarinic acid ultrafiltration clear liquid prepared in the step (2) into an ultrafilter with molecular weight cutoff of 500Da, and carrying out primary ultrafiltration under the condition that the gauge pressure is 0.5MPa until the volume ratio of the primary ultrafiltration cut-off liquid to the primary ultrafiltration filtrate is 1L: 4L. Respectively collecting the first ultrafiltration trapped fluid and the first ultrafiltration filtered fluid, pumping the collected first ultrafiltration filtered fluid into a biochemical tank for biochemical treatment, and discharging after reaching the standard; and adding purified water into the collected first ultrafiltration retentate for dilution, wherein the volume ratio of the first ultrafiltration retentate to the added purified water is 1L: 4L. And after the dilution is finished, carrying out second ultrafiltration until the volume ratio of the second ultrafiltration retentate to the second ultrafiltration filtrate is 1L: 4L, and stopping ultrafiltration. Respectively collecting the second ultrafiltration filtrate and the second ultrafiltration retentate, and collecting the second ultrafiltration retentate to obtain the rosmarinic acid ultrafiltration concentrate for preparing calcium rosmarinic acid; pumping the collected second ultrafiltration filtrate into a biochemical treatment tank for biochemical treatment, and discharging after reaching the standard. And (3) washing the collected ultrafiltration membrane with the molecular weight cutoff of 500Da after the second ultrafiltration separation for 20min by using purified water, then washing the ultrafiltration membrane for 20min by using 0.5% alkali liquor, and finally washing the ultrafiltration membrane by using the purified water until the pH value is 6 to obtain a regenerated ultrafiltration membrane, wherein the membrane can be used for preparing the next batch of rosmarinic acid ultrafiltration concentrated solution.
(4) Preparation of calcium Rosmarinate
Dissolving appropriate amount of calcium hydroxide in purified water, filtering, and collecting filtrate to obtain dilute calcium hydroxide solution containing Ca2+The mass percentage concentration is 0.5 percent and the pH value is 10. After the step (3) is finished, pumping the rosmarinic acid ultrafiltration concentrated solution prepared in the step (3) into a neutralization tank, controlling the stirring speed at 100r/min, and dropwise adding Ca2+And (3) diluting the calcium hydroxide solution with the mass percentage concentration of 0.5% and the pH value of 10 until obvious precipitation appears in the ultrafiltration concentrated solution, and stopping dropwise adding the diluted calcium hydroxide solution when the pH value reaches 10 to prepare the calcium rosmarinate suspension solution. Ca was added dropwise2+After completion of the dilute calcium hydroxide solution having a mass percentage concentration of 0.5% and a pH of 10, the stirring was stopped and the suspension was allowed to stand for 12 hours. Finally, filtering the suspension solution, respectively collecting filter residues and filtrate, and performing biochemical treatment on the collected filtrate to discharge after reaching the standard; and preparing the calcium rosmarinate from the collected filter residues for preparing a rosmarinic acid dissolving solution in the next step.
(5) Preparation of a solution of rosmarinic acid
And (4) after the step (4) is finished, dispersing the calcium rosmarinate prepared in the step (4) into purified water, wherein the mass ratio of the calcium rosmarinate to the volume of the purified water is 1 g: 500mL, adding the dilute hydrochloric acid solution used in the step (1) for preparing the raw rosmarinate solution under the condition that the stirring speed of a dissolving tank is 100r/min, and stopping dripping and stirring when the pH value of the system is 5. Then, filtration was performed until no filtrate flowed out. Collecting filtrate and filtering residue, and collecting filtrate to obtain rosmarinic acid solution for further preparation of rosmarinic acid eluate; and (4) using the collected filter residues for preparing the compound fertilizer.
(6) Preparation of Rosmarinic acid eluent
Soaking MCI GEL CHP 20 type resin with particle size of 75 μm in anhydrous ethanol 4 times the volume of resin (BV) for 8 hr, removing floating resin particles on the upper layer, filtering, and collecting residue and filtrate. Recovering ethanol from the collected filtrate; and balancing the collected filter residue with 4 times volume (BV) of ethanol aqueous solution with pH of 4 and volume percentage concentration of 60% for 2 hours, filling the filter residue into a column by a wet method, and connecting the column with a constant flow pump, an ultraviolet detector and an automatic fraction collector to form an MCI resin column separation system for later use.
And (5) after the step (5) is finished, pumping the rosmarinic acid dissolving solution prepared in the step (5) into a resin column separation system provided with an activated MCI GEL CHP 20 type resin with the particle size of 75 mu m, controlling the pumping flow rate of the rosmarinic acid dissolving solution to be 4 times of the volume (BV)/h of the resin, and controlling the volume ratio of the MCI resin to the dissolving solution to be 1L: 15L. And after the rosmarinic acid solution is pumped, pumping an ethanol water solution with the ethanol volume fraction of 80% for elution, wherein the pumping flow rate is 5BV/h, and respectively collecting eluates of the 4BV, the 8BV and the 12 BV. Combining the collected eluent of the 4 th BV and the eluent of the 12 th BV for recovering ethanol; and collecting the eluent of the 8BV, namely the rosmarinic acid eluent, and using the eluent to prepare rosmarinic acid powder in the next step. After the elution is finished, the column is washed by pumping an aqueous ethanol solution with the ethanol volume fraction of 90 percent, the volume of the aqueous ethanol solution is pumped to be 2BV, and the pumping flow rate is 1 BV/h. After the washing is completed, the MCI resin column can be used for the next batch to prepare the rosmarinic acid eluent.
(7) Preparation of Rosmarinic acid powder
And (6) after the step (6) is finished, pumping the rosmarinic acid eluent prepared in the step (6) into a rotary evaporator for reduced pressure concentration, distilling ethanol at the temperature of 70 ℃ and the relative vacuum degree of-0.06 MPa, stopping rotary evaporation when no condensed liquid flows out of a condenser, and respectively collecting rotary evaporation condensate and rotary evaporation concentrated liquid. And the rotary evaporation condensate can be recycled to an ethanol storage tank. And transferring the collected rotary evaporation concentrated solution into a vacuum drier for vacuum drying at 45 ℃ under the relative vacuum degree of-0.04 MPa for 6h to prepare rosmarinic acid powder. Wherein the content of rosmarinic acid is 91%, and the total yield of the rosmarinic acid powder is 88% of the mass of rosmarinic acid in the waste liquid generated in the process of producing the rosemary fat-soluble antioxidant.
Claims (1)
1. A method for extracting rosmarinic acid is characterized by comprising the following specific process steps:
(1) preparation of a Rosmarinic acid stock solution
According to the method introduced in the Chinese patent embodiment 1 with the publication number of CN 107739305A, preparing the centrifugal clear liquid in the step (2), namely the waste liquid generated in the process of producing the rosemary lipid-soluble antioxidant, uniformly mixing the waste liquid with purified water, wherein the volume ratio of the waste liquid to the purified water is 1L: 5-8L, adjusting the pH to 3-5 by using dilute hydrochloric acid, pumping the mixture into a tubular centrifuge, separating at the speed of 15000-20000 r/min, respectively collecting the centrifugal clear liquid and the centrifugal precipitate, and collecting the centrifugal clear liquid, namely the rosmarinic acid stock solution, for preparing the rosmarinic acid ultrafiltration clear liquid in the next step; the collected centrifugal precipitate is used for preparing the natural mildew-proof coating;
(2) preparation of Rosmarinic acid Ultrafiltration serum
After the step (1) is finished, pumping the rosmarinic acid stock solution prepared in the step (1) into an ultrafilter with the molecular weight cutoff of 10-50 KDa, carrying out primary ultrafiltration under the condition that the gauge pressure is 0.2-0.6 MPa until the volume ratio of the primary ultrafiltration cutoff solution to the filtrate is 1L: 3-5L, respectively collecting the primary ultrafiltration cutoff solution and the primary ultrafiltration filtrate, and combining the collected primary ultrafiltration filtrate with the filtrate obtained by the subsequent secondary ultrafiltration for preparing the rosmarinic acid ultrafiltration concentrated solution in the next step; adding purified water into the collected first ultrafiltration trapped fluid, carrying out second ultrafiltration, wherein the volume ratio of the first ultrafiltration trapped fluid to the added purified water is 1L: 3-5L, the second ultrafiltration gauge pressure is 0.2-0.6 MPa, stopping ultrafiltration until the volume ratio of the second ultrafiltration trapped fluid to the filtrate thereof is 1L: 3-5L, respectively collecting the second ultrafiltration filtrate, the second ultrafiltration trapped fluid and the ultrafiltration membrane with the molecular weight cutoff of 10-50 KDa after the second ultrafiltration separation, washing for 15-20 min with purified water, then washing for 15-20 min with 0.5-1% alkali liquor, and finally washing with purified water until the pH value is 5-7 to obtain a regenerated ultrafiltration membrane, wherein the membrane can be used for preparing the next batch of rosmarinic acid ultrafiltration clear liquid;
collecting the second ultrafiltration filtrate, mixing with the first ultrafiltration filtrate to obtain rosmarinic acid ultrafiltration clear solution, and pumping the second ultrafiltration retentate mainly containing macromolecular polysaccharide and protein into biochemical tank for biochemical treatment, and discharging;
(3) preparation of Rosmarinic acid ultrafiltered concentrate
After the step (2) is finished, pumping the rosmarinic acid ultrafiltration clear liquid prepared in the step (2) into an ultrafilter with molecular weight cutoff of 500-1000 Da, carrying out primary ultrafiltration under the condition that gauge pressure is 0.2-0.6 MPa until the volume ratio of the primary ultrafiltration trapped liquid to the primary ultrafiltration filtered liquid is 1L: 3-5L, respectively collecting the primary ultrafiltration trapped liquid and the primary ultrafiltration filtered liquid, pumping the collected primary ultrafiltration filtered liquid into a biochemical tank for biochemical treatment, and discharging after reaching the standard; adding purified water into the collected first ultrafiltration trapped fluid for dilution, wherein the volume ratio of the first ultrafiltration trapped fluid to the added purified water is 1L: 3-5L, carrying out second ultrafiltration after dilution is finished until the volume ratio of the second ultrafiltration trapped fluid to the second ultrafiltration filtered fluid is 1L: 3-5L, stopping ultrafiltration, respectively collecting the second ultrafiltration trapped fluid and the second ultrafiltration trapped fluid, and obtaining rosmarinic acid ultrafiltration concentrated solution from the collected second ultrafiltration trapped fluid for the next step of preparing calcium rosmarinic acid; pumping the collected second ultrafiltration filtrate into a biochemical treatment tank for biochemical treatment, discharging after reaching the standard, washing the ultrafiltration membrane with the molecular weight cutoff of 500-1000 Da after the collected second ultrafiltration separation with purified water for 15-20 min, then washing with 0.5-1% alkali liquor for 15-20 min, and finally washing with purified water until the pH value is 5-7 to obtain a regenerated ultrafiltration membrane, wherein the membrane can be used for preparing the next batch of rosmarinic acid ultrafiltration concentrated solution;
(4) preparation of calcium Rosmarinate
Dissolving appropriate amount of calcium hydroxide in purified water, filtering, and collecting filtrate to obtain dilute calcium hydroxide solution containing Ca2+The mass percentage concentration is 0.1-0.5%, the pH value is 8-10, after the step (3) is finished, the rosmarinic acid ultrafiltration concentrated solution prepared in the step (3) is pumped into a neutralization tank, the stirring speed is controlled to be 50-100 r/min, and Ca is dropwise added2+Dilute calcium hydroxide solution with mass percentage concentration of 0.1-0.5% and pH of 8-10, stopping dropwise adding the dilute calcium hydroxide solution until obvious precipitation appears in the ultrafiltration concentrated solution and the pH value reaches 8-10, thus preparing a rosmarinic acid calcium suspension solution, and dropwise adding Ca2+Stopping stirring and standing the suspension solution for 6-12 hours after the completion of the dilute calcium hydroxide solution with the mass percentage concentration of 0.1-0.5% and the pH of 8-10, finally filtering the suspension solution, respectively collecting filter residue and filtrate, performing biochemical treatment on the collected filtrate, and discharging after reaching the standard; preparing calcium rosmarinci acid from the collected filter residue, and using the calcium rosmarinci acid to prepare a rosmarinci acid solution in the next step;
(5) preparation of a solution of rosmarinic acid
After the step (4) is finished, dispersing the calcium rosmarinate prepared in the step (4) into purified water, wherein the mass ratio of the calcium rosmarinate to the volume ratio of the purified water is 1 g: 500-1000 mL, adding the dilute hydrochloric acid solution used in the raw rosmarinate prepared in the step (1) under the condition that the stirring speed of a dissolving tank is 50-100 r/min, stopping dripping and stirring when the pH value of a system is 4-6, then filtering until no filtrate flows out, respectively collecting filtrate and filtering residues, and preparing rosmarinic acid dissolving liquid for next step of preparing rosmarinic acid eluent; the collected filter residues are used for preparing the compound fertilizer;
(6) preparation of Rosmarinic acid eluent
Soaking MCI GEL CHP 20 type resin with the particle size specification of 35-75 mu m in absolute ethyl alcohol with the volume (BV) of 4-6 times of that of the resin for 4-8 hours, removing resin particles floating on the upper layer, filtering, respectively collecting filter residue and filtrate, and recovering the ethanol from the collected filtrate; balancing the collected filter residues with an ethanol water solution with 4-6 times volume (BV), pH of 3-5 and volume percentage concentration of 40-60% for 1-2 hours, then filling the filter residues into a column by a wet method, and connecting the column with a constant flow pump, an ultraviolet detector and an automatic fraction collector to form an MCI resin column separation system for later use;
after the step (5) is finished, pumping the rosmarinic acid dissolving solution prepared in the step (5) into an activated MCI GEL CHP 20 type resin column separation system with the particle size of 35-75 microns, controlling the pumping flow rate of the rosmarinic acid dissolving solution to be 2-5 times of the volume (BV)/h of the resin, controlling the ratio of the volume of the MCI resin to the volume of the dissolving solution to be 1L: 10-15L, after the rosmarinic acid dissolving solution is pumped, pumping an ethanol water solution with the volume fraction of 70-90% of ethanol for elution, wherein the pumping flow rate is 1-5 BV/h, respectively collecting eluents of 1-4 BV, 5-8 BV and 9-12 BV, and combining the collected eluents of 1-4 BV and 9-12 BV for recovering ethanol; the collected eluent of 5-8 BV is rosmarinic acid eluent, and is used for preparing rosmarinic acid powder in the next step, after elution is completed, an ethanol water solution with the ethanol volume fraction of 90-95% is pumped in for washing, the volume of the ethanol water solution is pumped in for 1-6 BV, the pumping flow rate is 1-5 BV/h, and the washed MCI resin column can be used for preparing rosmarinic acid eluent in the next step;
(7) preparation of Rosmarinic acid powder
And (3) after the step (6) is finished, pumping the rosmarinic acid eluent prepared in the step (6) into a rotary evaporator for reduced pressure concentration, distilling ethanol at the temperature of 60-70 ℃ and the relative vacuum degree of-0.06-0.09 MPa, stopping reduced pressure concentration when no condensed liquid flows out of a condenser, respectively collecting a condensate and a concentrated solution, recovering ethanol from the collected condensate, transferring the collected concentrated solution into a vacuum drier, and performing vacuum drying for 5-8 hours under the conditions that the temperature is 45-50 ℃ and the relative vacuum degree is-0.04-0.08 MPa to prepare rosmarinic acid powder, wherein the content of rosmarinic acid is 89-98%, and the total yield of the rosmarinic acid powder is 85-93% of the mass of rosmarinic acid in waste liquid generated in the process of producing the rosmarinic acid fat-soluble antioxidant.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101863767A (en) * | 2010-03-23 | 2010-10-20 | 南京泽朗农业发展有限公司 | Preparation process of rosmarinic acid |
| CN103044253A (en) * | 2013-01-05 | 2013-04-17 | 青海伊纳维康生物科技有限公司 | Extraction separation method of rosmarinic acid in salvia castabea diels f. tomentosa stib |
| CN107739305A (en) * | 2017-09-11 | 2018-02-27 | 湖南鑫利生物科技有限公司 | A kind of method for improving carnosic acid content in rosemary fat-soluble antioxidant |
| CN108558670A (en) * | 2018-06-01 | 2018-09-21 | 湖南鑫利生物科技有限公司 | A method of preparing Rosmarinic acid |
| WO2021038340A1 (en) * | 2019-08-26 | 2021-03-04 | Tübi̇tak | Carnosic acid, carnosol and rosmarinic acid isolation method |
-
2021
- 2021-06-09 CN CN202110668807.2A patent/CN113461531A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101863767A (en) * | 2010-03-23 | 2010-10-20 | 南京泽朗农业发展有限公司 | Preparation process of rosmarinic acid |
| CN103044253A (en) * | 2013-01-05 | 2013-04-17 | 青海伊纳维康生物科技有限公司 | Extraction separation method of rosmarinic acid in salvia castabea diels f. tomentosa stib |
| CN107739305A (en) * | 2017-09-11 | 2018-02-27 | 湖南鑫利生物科技有限公司 | A kind of method for improving carnosic acid content in rosemary fat-soluble antioxidant |
| CN108558670A (en) * | 2018-06-01 | 2018-09-21 | 湖南鑫利生物科技有限公司 | A method of preparing Rosmarinic acid |
| WO2021038340A1 (en) * | 2019-08-26 | 2021-03-04 | Tübi̇tak | Carnosic acid, carnosol and rosmarinic acid isolation method |
Non-Patent Citations (1)
| Title |
|---|
| 张海龙: "高纯度迷迭香酸的分离纯化工艺研究", 硕士学位论文工程科技I辑, no. 04, pages 1 - 47 * |
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