CN103319495A - Preparation method of high-purity carnosol - Google Patents
Preparation method of high-purity carnosol Download PDFInfo
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- CN103319495A CN103319495A CN2013103050811A CN201310305081A CN103319495A CN 103319495 A CN103319495 A CN 103319495A CN 2013103050811 A CN2013103050811 A CN 2013103050811A CN 201310305081 A CN201310305081 A CN 201310305081A CN 103319495 A CN103319495 A CN 103319495A
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- carnosol
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- elutriant
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- QRYRORQUOLYVBU-VBKZILBWSA-N Carnosic acid Natural products 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 title claims abstract description 104
- 235000004654 carnosol Nutrition 0.000 title claims abstract description 89
- XUSYGBPHQBWGAD-PJSUUKDQSA-N Carnosol Chemical compound CC([C@@H]1C2)(C)CCC[C@@]11C(=O)O[C@@H]2C2=C1C(O)=C(O)C(C(C)C)=C2 XUSYGBPHQBWGAD-PJSUUKDQSA-N 0.000 title claims abstract description 88
- MMFRMKXYTWBMOM-UHFFFAOYSA-N Carnosol Natural products CCc1cc2C3CC4C(C)(C)CCCC4(C(=O)O3)c2c(O)c1O MMFRMKXYTWBMOM-UHFFFAOYSA-N 0.000 title claims abstract description 88
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 239000003960 organic solvent Substances 0.000 claims abstract description 27
- 239000000284 extract Substances 0.000 claims abstract description 20
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 16
- 239000012043 crude product Substances 0.000 claims abstract description 15
- 239000011347 resin Substances 0.000 claims abstract description 13
- 229920005989 resin Polymers 0.000 claims abstract description 13
- 239000002994 raw material Substances 0.000 claims abstract description 8
- 239000013078 crystal Substances 0.000 claims abstract description 5
- 239000007787 solid Substances 0.000 claims abstract description 5
- 238000001514 detection method Methods 0.000 claims abstract description 4
- 239000000706 filtrate Substances 0.000 claims abstract description 3
- 238000001556 precipitation Methods 0.000 claims description 25
- 238000002425 crystallisation Methods 0.000 claims description 19
- 230000008025 crystallization Effects 0.000 claims description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 12
- 239000003463 adsorbent Substances 0.000 claims description 10
- 235000015639 rosmarinus officinalis Nutrition 0.000 claims description 10
- 241000196324 Embryophyta Species 0.000 claims description 9
- 238000009736 wetting Methods 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 7
- 238000000605 extraction Methods 0.000 claims description 7
- 238000000151 deposition Methods 0.000 claims description 5
- 238000009835 boiling Methods 0.000 claims description 4
- 235000008504 concentrate Nutrition 0.000 claims description 4
- 239000012141 concentrate Substances 0.000 claims description 4
- 244000295490 Salvia japonica Species 0.000 claims description 2
- 235000005794 Salvia japonica Nutrition 0.000 claims description 2
- 239000000796 flavoring agent Substances 0.000 claims description 2
- 235000019634 flavors Nutrition 0.000 claims description 2
- 235000014666 liquid concentrate Nutrition 0.000 claims description 2
- 241001530490 Salvia rosmarinus Species 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 15
- 238000001914 filtration Methods 0.000 abstract description 8
- 150000001875 compounds Chemical class 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 239000002244 precipitate Substances 0.000 abstract 4
- 230000001376 precipitating effect Effects 0.000 abstract 2
- 238000007670 refining Methods 0.000 abstract 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 14
- 244000178231 Rosmarinus officinalis Species 0.000 description 9
- LFEINUNSYODISY-UHFFFAOYSA-N (ent-5alpha,6beta)-15,16-Epoxy-3,13(16),14-clerodatrien-18,6-olide Natural products CC1CC(C23C)OC(=O)C3=CCCC2C1(C)CCC=1C=COC=1 LFEINUNSYODISY-UHFFFAOYSA-N 0.000 description 8
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 8
- 239000003921 oil Substances 0.000 description 8
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 7
- 229960004756 ethanol Drugs 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 239000012452 mother liquor Substances 0.000 description 5
- 239000008213 purified water Substances 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 241001529742 Rosmarinus Species 0.000 description 4
- 238000004090 dissolution Methods 0.000 description 4
- 238000001179 sorption measurement Methods 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 235000006708 antioxidants Nutrition 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 229940092258 rosemary extract Drugs 0.000 description 3
- 235000020748 rosemary extract Nutrition 0.000 description 3
- 239000001233 rosmarinus officinalis l. extract Substances 0.000 description 3
- 238000005303 weighing Methods 0.000 description 3
- 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 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 229960000935 dehydrated alcohol Drugs 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 238000005502 peroxidation Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- 238000010898 silica gel chromatography Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000000341 volatile oil Substances 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 1
- 102000003945 NF-kappa B Human genes 0.000 description 1
- 108010057466 NF-kappa B Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 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 description 1
- 240000007164 Salvia officinalis Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000000118 anti-neoplastic effect Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 150000004965 peroxy acids Chemical class 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 235000005412 red sage Nutrition 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 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 description 1
- 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 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000000194 supercritical-fluid extraction Methods 0.000 description 1
- 239000013076 target substance Substances 0.000 description 1
Landscapes
- Medicines Containing Plant Substances (AREA)
- Saccharide Compounds (AREA)
Abstract
The invention relates to a preparation method of a natural active compound. The preparation method provided by the invention comprises the following steps: 1, extracting a dry carnosol-containing raw material per kilogram by the use of 3-10 liters of a lipophilic organic solvent for 2-5 times, merging extracts, and condensing to obtain a carnosol-containing extract or solid; 2, dissolving by the use of a 10-90% hydrophilic organic solvent, filtering and precipitating, and carrying out macroreticular resin refining processing on a filtrate to obtain a carnosol-containing precipitate; 3, dissolving the precipitate by the use of a mixed lipophilic organic solvent, standing, precipitating a precipitate; and filtering the precipitate to obtain a carnosol crude product; and 4, crystallizing by the use of a hydrophilic organic solvent so as to obtain high-purity carnosol crystals with the HPLC detection content being more than 95%. The preparation method provided by the invention has advantages of simple operation process and controllable cost, and is suitable for large-scale preparation and production of high-purity carnosol.
Description
Technical field
The present invention relates to a kind of preparation method of natural radioactivity compound.
Background technology
Rosmarinus officinalis (
Rosmarinus officinalisL.) etc. the phenol diterpene-kind compounds such as carnosol that contain in the plant are the natural antioxidantss of generally acknowledging in recent years, are widely used in fields such as foodstuffs industry, day chemical industry and medicines and health protection; And studies show that carnosol can have antineoplastic action by suppressing the NF-kB passage, so it is a new drug research person natural radioactivity compound looking at of parent relatively in recent years.
About the preparation of carnosol, among the Chinese patent CN101040901A, by Rosmarinus officinalis is put forward methods such as taking, putting cryoprecipitate, activated carbon decolorizing with solvent, make the rosemary extract that contains carnosol 5-12%; Among the Chinese patent CN101402864A, by to Rosmarinus officinalis earlier with the vapor extraction essential oil, organic solvent extracts processes such as residue, macroporous adsorbent resin and silica gel column chromatography again, obtains containing carnosol and the rosemary antioxidant of Salvin sum more than 50%; Among the Chinese patent CN102031116A, by to Rosmarinus officinalis earlier with the vapor extraction essential oil, again organic solvent extract residue obtain extract, with CO
2This extract of supercritical extraction obtains containing the main rosemary antioxidant of oxidation-resistant active ingredient more than 80% such as carnosol, Salvin, rosmarinic acid.The product that these methods obtain all is the mixture that contains a certain amount of carnosol, and for research and the application of a monomer reactivity material, these mixtures do not meet standard and requirement.Among the Chinese patent CN102115442A; by Rosmarinus officinalis is extracted, carries out means such as phosphoric acid-silica gel column chromatography under nitrogen protection with solvent; obtain content and reach 92% carnosol; though this method can obtain the carnosol of higher degree, used condition is too harsh, and silica gel is wanted phosphorylation; make phosphoric acid-silica gel; the nitrogen protection of chromatography process need is difficult to carry out a large amount of preparation carnosols, also is difficult to promote and this special methods of practical application.Restrain sharp David Stauffer Wei Ruili at patent CN101835782A, CN101835783A, and among the CN101835783A, means such as the oxidation by Salvin being carried out hydrogen peroxide or peroxy acid, acetic acid crystallization obtain the carnosol more than 90%.This method gained carnosol content height, flow process is fairly simple, and is better than above-mentioned other several method.But needing highly purified Salvin is raw material; Salvin also is the active compound that extraction separation obtains from Rosmarinus officinalis etc. plant; obtain highly purified carnosol if carry out peroxidation with highly purified Salvin; then its source will be limited greatly; and manufacturing cost is very high; if make carnosol with the Salvin that purity is not high; the carnosol yield that then prepares is low; be difficult to obtain a large amount of target substances; therefore, obtain the method for highly purified carnosol with the Salvin peroxidation, aspect mass-producing and cost control; very big difficulty is arranged, be difficult to practical application.
Summary of the invention
The objective of the invention is at preparation high purity carnosol in the prior art exist not enough and difficult, propose that a kind of flow process is simple, cost is controlled, be fit to the preparation method of the high purity carnosol of large-scale production.
The preparation method of high purity carnosol of the present invention is made up of following steps:
One, with the raw material that contains carnosol of drying, each kilogram rises the lipotropy organic solvent with 3-10 and extracts 2-5 time, and united extraction liquid concentrates the medicinal extract or the solid that obtain containing carnosol;
Two, the described medicinal extract of each kilogram or solid, rise the wetting ability organic solvent dissolving of 10-90% again with 5-10, filtering-depositing, filtrate is carried out the macroporous adsorbent resin refinement treatment, the elutriant Fractional Collections, merge carnosol content greater than the total elutriant of elutriant conduct of 4g/L, concentrate total elutriant to solvent-free flavor; Add the concentrated solution volume again than 1-3 water doubly, stir, precipitation, filter the throw out of carnosol;
Three, the described throw out of each kilogram rises the lipotropy organic solvent dissolving that mixes with 5-10 again, places, and separates out precipitation; Filtering-depositing gets the carnosol crude product;
Four, the described carnosol crude product of each kilogram with the wetting ability organic solvent crystallization of 8-12 liter, obtains with the HPLC detection level at the high purity carnosol crystal more than 95% again.
The described raw material that contains carnosol of step 1, refer to Rosmarinus officinalis (
Rosmarinus officinalisL.), Salvia japonica Thunb., the red sage root etc. contain the plant of carnosol, wherein contain carnosol and are not less than 0.1% to detect HPLC, or the extract of these plants, wherein contain carnosol and be not less than 1% to detect HPLC.
The described lipotropy organic solvent of step 1, refer to boiling point be lower than 100 degrees centigrade, with the organic solvent that layering can take place after water mixes, as sherwood oil, benzene, ether, methylene dichloride, chloroform, ethyl acetate etc.
The described wetting ability organic solvent of step 2 refers to that boiling point is lower than 100 degrees centigrade, organic solvent that can be miscible with water, as acetone, ethanol, methyl alcohol, Virahol etc.Described 10-90% is the volume percent aqueous solution.More superior is the 40-70% volume percent aqueous solution.
The described macroporous adsorbent resin of step 2 refers to wherein any one of D101, AB-8, HZ818.
The present invention adopts the lipotropy organic solvent that the raw material that contains carnosol is extracted, and fat-soluble component is dissolved in the organic solvent, has fully effectively reduced the interference of big polar material such as polysaccharide, protein in the plant; And then will extract concentrated solution wetting ability organic solvent dissolving, filtering-depositing is removed with carnosol polarity and is differed bigger little polar material, has been further purified macroporous resin upper prop liquid, attached with parsing by the macroporous resin selective adsorption, Fractional Collections contains the carnosol lower column liquid; Use lipotropy organic solvent and the fractional precipitation of wetting ability organic solvent and crystallization treatment again, both got the high purity carnosol crystal more than 95%.
The method of the invention can be from the lower raw material of carnosol content, and the disposable carnosol content that obtains is higher than 95% high purity carnosol crystal, is that this field can not be accomplished at present.Present method operating process is simple, cost is controlled, preparation and the production high purity carnosol of suitable mass-producing.
Embodiment
Embodiment 1:
Get dry Rosmarinus officinalis branches and leaves 2000g, add sherwood oil 14L, refluxing extraction 3 times, each 3 hours, merge No. 3 times extracting solution, concentrate and reclaim sherwood oil, get medicinal extract; With medicinal extract with 40% dissolve with ethanol, remove by filter insolubles, get the 1000ml mother liquor as adsorption liquid, adsorb with 500ml D101 macroporous adsorbent resin, after finishing, absorption leads to wash-out with the 1000ml purified water, use 1500ml 90% ethanolic soln wash-out again, elutriant is divided into 6 sections, merges middle 4 parts; The elutriant that merges is evaporated to 100ml, adds water 200ml, stirs and separates out precipitation, places 3 hours; Centrifugal the precipitation that contains carnosol; Precipitation 200ml acetic acid ethyl dissolution to wherein adding the 200ml sherwood oil, stirs, and separates out precipitation; Filter the carnosol crude product, this crude product obtains the carnosol crystallization with the crystallization of 800ml dehydrated alcohol; Filtration, drying obtain crystallization 13.6 g, and it is 96.1% that HPLC detects carnosol content.
Embodiment 2:
Get dry Rosmarinus officinalis branches and leaves 2000g, add methylene dichloride 10L, refluxing extraction 3 times, each 3 hours, merge No. 3 times extracting solution, concentrate and reclaim methylene dichloride, get medicinal extract; With medicinal extract with 60% dissolve with ethanol, remove by filter insolubles, get the 1000ml mother liquor as adsorption liquid, adsorb with 500ml AB-8 macroporous adsorbent resin, after finishing, absorption leads to wash-out with the 1000ml purified water, use 2000ml 80% ethanolic soln wash-out again, elutriant is divided into 6 sections, merges middle 4 parts; The elutriant that merges is evaporated to 150ml, adds water 200ml, stirs and separates out precipitation, places 5 hours; Centrifugal the precipitation that contains carnosol; Precipitation, stirs to wherein adding the 200ml sherwood oil with the dissolving of 150ml methylene dichloride, separates out precipitation; Filter the carnosol crude product, this crude product obtains the carnosol crystallization with the 700ml methanol crystallization; Filtration, drying obtain crystallization 14.7 g, and it is 95.3% that HPLC detects carnosol content.
Embodiment 3:
Taking by weighing rosemary extract 500g(HPLC mensuration carnosol content is 5.7%), dissolve with ethanol with 2500ml 60%, remove by filter insolubles, mother liquor is as adsorption liquid, adsorb with 800ml HZ818 macroporous adsorbent resin, with the logical wash-out of 2000ml purified water, use 2000ml 90% ethanolic soln wash-out again after absorption is finished, elutriant is divided into 6 sections, merges middle 4 parts; The elutriant that merges is evaporated to 300ml, adds water 350ml, stirs and separates out precipitation, places 3 hours; Centrifugal the precipitation that contains carnosol; Precipitation 300ml acetic acid ethyl dissolution to wherein adding the 200ml sherwood oil, stirs, and separates out precipitation; Filter the carnosol crude product, this crude product obtains the carnosol crystallization with the 1000ml methanol crystallization; Filtration, drying obtain crystallization 23.9 g, and it is 96.3% that HPLC detects carnosol content.
Embodiment 4:
Taking by weighing Radix Salviae Miltiorrhizae extract 500g(HPLC mensuration carnosol content is 3.1%), dissolve with ethanol with 3000ml 50%, remove by filter insolubles, mother liquor adsorbs with 1000ml D101 macroporous adsorbent resin, after finishing, absorption leads to wash-out with the 2000ml purified water, use 2000ml 90% ethanolic soln wash-out again, elutriant is divided into 6 sections, merges middle 4 parts; The elutriant that merges is evaporated to 500ml, adds water 500ml, stirs and separates out precipitation, places 3 hours; Centrifugal the precipitation that contains carnosol; Precipitation 200ml acetic acid ethyl dissolution to wherein adding the 150ml sherwood oil, stirs, and separates out precipitation; Filter the carnosol crude product, this crude product obtains the carnosol crystallization with the crystallization of 600ml dehydrated alcohol; Filtration, drying obtain crystallization 12.2 g, and it is 95.8% that HPLC detects carnosol content.
Embodiment 5:
Taking by weighing rosemary extract 500g(HPLC mensuration carnosol content is 17.0%), dissolve with ethanol with 4000ml 60%, remove by filter insolubles, mother liquor adsorbs with 1500ml D101 macroporous adsorbent resin, after finishing, absorption leads to wash-out with the 4000ml purified water, use 4000ml 90% ethanolic soln wash-out again, elutriant is divided into 6 sections, merges middle 4 parts; The elutriant that merges is evaporated to 800ml, adds water 1000ml, stirs and separates out precipitation, places 4 hours; Centrifugal the precipitation that contains carnosol; Precipitation 1000ml acetic acid ethyl dissolution to wherein adding the 800ml sherwood oil, stirs, and separates out precipitation; Filter the carnosol crude product, this crude product obtains the carnosol crystallization with the 4500ml methanol crystallization; Filtration, drying obtain crystallization 73.3 g, and it is 96.8% that HPLC detects carnosol content.
The HPLC detection method:
Test set: Tianjin, island-LC-10AT vp plus
Chromatographic column: Wondasil
TMC18 5nm 4.6*250mm
Chromatographic condition:
Mobile phase A: 0.1% phosphate aqueous solution (crossing the 0.45nm filtering membrane)
Mobile phase B: acetonitrile (chromatographic grade)
A：B=40：60
Flow velocity: 0.8ml/mine
Detect wavelength: 285nm
Appearance time: about 14mine
Trial-product and reference substance obtain solution: hplc grade methanol dissolving.
Claims (7)
1. the preparation method of a high purity carnosol is characterized in that being made up of following steps:
One, with the raw material that contains carnosol of drying, each kilogram rises the lipotropy organic solvent with 3-10 and extracts 2-5 time, and united extraction liquid concentrates the medicinal extract or the solid that obtain containing carnosol;
Two, the described medicinal extract of each kilogram or solid, rise the wetting ability organic solvent dissolving of 10-90% again with 5-10, filtering-depositing, filtrate is carried out the macroporous adsorbent resin refinement treatment, the elutriant Fractional Collections, merge carnosol content greater than the total elutriant of elutriant conduct of 4g/L, concentrate total elutriant to solvent-free flavor; Add the concentrated solution volume again than 1-3 water doubly, stir, precipitation, filter the throw out of carnosol;
Three, the described throw out of each kilogram rises the lipotropy organic solvent dissolving that mixes with 5-10 again, places, and separates out precipitation; Filtering-depositing gets the carnosol crude product;
Four, the described carnosol crude product of each kilogram with the wetting ability organic solvent crystallization of 8-12 liter, obtains with the HPLC detection level at the high purity carnosol crystal more than 95% again.
2. the preparation method of high purity carnosol as claimed in claim 1, it is characterized in that the described raw material that contains carnosol of step (), refer to contain the plant of carnosol, wherein contain carnosol and be not less than 0.1% to detect HPLC, or the extract of these plants, wherein contain carnosol and be not less than 1% to detect HPLC.
3. the preparation method of high purity carnosol as claimed in claim 1 is characterized in that the described lipotropy organic solvent of step (), refer to boiling point be lower than 100 degrees centigrade, with the organic solvent that layering can take place after water mixes.
4. the preparation method of high purity carnosol as claimed in claim 1 is characterized in that the described wetting ability organic solvent of step (two), refers to that boiling point is lower than 100 degrees centigrade, organic solvent that can be miscible with water.
5. the preparation method of high purity carnosol as claimed in claim 1 is characterized in that the described macroporous adsorbent resin of step (two), refers to wherein any one of D101, AB-8, HZ818.
6. the preparation method of high purity carnosol as claimed in claim 2 is characterized in that the described plant that contains carnosol is Rosmarinus officinalis.
7. the preparation method of high purity carnosol as claimed in claim 2 is characterized in that the described plant that contains carnosol is Salvia japonica Thunb..
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104530074A (en) * | 2015-01-07 | 2015-04-22 | 浙江泰康生物科技有限公司 | Method for extracting carnosol from complete salvia |
| CN105696194A (en) * | 2016-04-05 | 2016-06-22 | 东华大学 | Preparation method of carnosol and chitosan self-assembly core sheath composite nanometer fiber mat |
| CN105696195A (en) * | 2016-04-05 | 2016-06-22 | 东华大学 | Preparation method of carnosol and chitosan composite nanometer fiber mat |
| CN111440184A (en) * | 2018-01-30 | 2020-07-24 | 北京联合大学 | Method for preparing high-purity carnosol |
| CN114988983A (en) * | 2022-06-09 | 2022-09-02 | 新疆师范大学 | Method for separating sclareol from sclareol extract |
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| TR201912820A2 (en) * | 2019-08-26 | 2021-03-22 | Tuerkiye Bilimsel Ve Teknolojik Arastirma Kurumu Tuebitak | CARNOSIC ACID, CARNOSOL AND ROSMARINIC ACID ISOLATION METHOD |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104530074A (en) * | 2015-01-07 | 2015-04-22 | 浙江泰康生物科技有限公司 | Method for extracting carnosol from complete salvia |
| CN105696194A (en) * | 2016-04-05 | 2016-06-22 | 东华大学 | Preparation method of carnosol and chitosan self-assembly core sheath composite nanometer fiber mat |
| CN105696195A (en) * | 2016-04-05 | 2016-06-22 | 东华大学 | Preparation method of carnosol and chitosan composite nanometer fiber mat |
| CN111440184A (en) * | 2018-01-30 | 2020-07-24 | 北京联合大学 | Method for preparing high-purity carnosol |
| CN111440184B (en) * | 2018-01-30 | 2021-08-27 | 北京联合大学 | Method for preparing high-purity carnosol |
| CN114988983A (en) * | 2022-06-09 | 2022-09-02 | 新疆师范大学 | Method for separating sclareol from sclareol extract |
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