CN103073417B - The preparation method of high-purity trans-crocetin - Google Patents
The preparation method of high-purity trans-crocetin Download PDFInfo
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- CN103073417B CN103073417B CN201310066851.1A CN201310066851A CN103073417B CN 103073417 B CN103073417 B CN 103073417B CN 201310066851 A CN201310066851 A CN 201310066851A CN 103073417 B CN103073417 B CN 103073417B
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- 229950004497 transcrocetin Drugs 0.000 title claims abstract description 126
- PANKHBYNKQNAHN-MQQNZMFNSA-N crocetin Chemical compound OC(=O)C(/C)=C/C=C/C(/C)=C/C=C/C=C(\C)/C=C/C=C(\C)C(O)=O PANKHBYNKQNAHN-MQQNZMFNSA-N 0.000 title claims abstract description 104
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims abstract description 109
- -1 trans-crocetin pyridinium salt Chemical class 0.000 claims abstract description 63
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims abstract description 54
- PANKHBYNKQNAHN-JTBLXSOISA-N Crocetin Natural products OC(=O)C(\C)=C/C=C/C(/C)=C\C=C\C=C(\C)/C=C/C=C(/C)C(O)=O PANKHBYNKQNAHN-JTBLXSOISA-N 0.000 claims abstract description 38
- PANKHBYNKQNAHN-JUMCEFIXSA-N carotenoid dicarboxylic acid Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C(=O)O)C=CC=C(/C)C(=O)O PANKHBYNKQNAHN-JUMCEFIXSA-N 0.000 claims abstract description 38
- 239000012043 crude product Substances 0.000 claims abstract description 31
- 238000001953 recrystallisation Methods 0.000 claims abstract description 24
- 238000002425 crystallisation Methods 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 14
- 230000008025 crystallization Effects 0.000 claims abstract description 13
- 239000000243 solution Substances 0.000 claims description 35
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 32
- 238000001556 precipitation Methods 0.000 claims description 32
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 30
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 24
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 24
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 22
- 239000007864 aqueous solution Substances 0.000 claims description 21
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 18
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 230000007935 neutral effect Effects 0.000 claims description 17
- 238000005406 washing Methods 0.000 claims description 17
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 16
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 16
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 16
- 239000003960 organic solvent Substances 0.000 claims description 15
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 14
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 13
- 239000002253 acid Substances 0.000 claims description 13
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 claims description 12
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 9
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 8
- 230000002378 acidificating effect Effects 0.000 claims description 8
- 238000004090 dissolution Methods 0.000 claims description 8
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 6
- 230000002209 hydrophobic effect Effects 0.000 claims description 6
- ZXEKIIBDNHEJCQ-UHFFFAOYSA-N isobutanol Chemical compound CC(C)CO ZXEKIIBDNHEJCQ-UHFFFAOYSA-N 0.000 claims description 6
- 239000012046 mixed solvent Substances 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 5
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 4
- 239000003513 alkali Substances 0.000 claims description 4
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 3
- 239000000178 monomer Substances 0.000 claims description 3
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 claims description 3
- 229920002554 vinyl polymer Polymers 0.000 claims description 3
- XVMSFILGAMDHEY-UHFFFAOYSA-N 6-(4-aminophenyl)sulfonylpyridin-3-amine Chemical compound C1=CC(N)=CC=C1S(=O)(=O)C1=CC=C(N)C=N1 XVMSFILGAMDHEY-UHFFFAOYSA-N 0.000 claims description 2
- 235000005979 Citrus limon Nutrition 0.000 claims description 2
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 2
- 239000011260 aqueous acid Substances 0.000 claims description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Substances [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 2
- 235000015320 potassium carbonate Nutrition 0.000 claims description 2
- 235000011118 potassium hydroxide Nutrition 0.000 claims description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 2
- 235000017550 sodium carbonate Nutrition 0.000 claims description 2
- 235000011121 sodium hydroxide Nutrition 0.000 claims description 2
- 244000248349 Citrus limon Species 0.000 claims 1
- 239000002904 solvent Substances 0.000 abstract description 19
- 239000000047 product Substances 0.000 abstract description 5
- 238000000926 separation method Methods 0.000 abstract description 3
- 229910052736 halogen Inorganic materials 0.000 abstract description 2
- 150000002367 halogens Chemical class 0.000 abstract description 2
- 241000411851 herbal medicine Species 0.000 abstract description 2
- 230000003321 amplification Effects 0.000 abstract 1
- 238000003199 nucleic acid amplification method Methods 0.000 abstract 1
- 239000013078 crystal Substances 0.000 description 28
- 238000004128 high performance liquid chromatography Methods 0.000 description 23
- 238000001291 vacuum drying Methods 0.000 description 13
- 239000012535 impurity Substances 0.000 description 12
- 239000000706 filtrate Substances 0.000 description 10
- 239000007788 liquid Substances 0.000 description 8
- SEBIKDIMAPSUBY-ARYZWOCPSA-N Crocin Chemical compound C([C@H]1O[C@H]([C@@H]([C@@H](O)[C@@H]1O)O)OC(=O)C(C)=CC=CC(C)=C\C=C\C=C(/C)\C=C\C=C(C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)O1)O)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O SEBIKDIMAPSUBY-ARYZWOCPSA-N 0.000 description 7
- 239000007787 solid Substances 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 5
- 238000003860 storage Methods 0.000 description 5
- 235000015655 Crocus sativus Nutrition 0.000 description 4
- 244000124209 Crocus sativus Species 0.000 description 4
- 229960000583 acetic acid Drugs 0.000 description 4
- 239000009627 gardenia yellow Substances 0.000 description 4
- 150000003839 salts Chemical class 0.000 description 4
- 235000018958 Gardenia augusta Nutrition 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- SEBIKDIMAPSUBY-JAUCNNNOSA-N Crocin Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C(=O)OC1OC(COC2OC(CO)C(O)C(O)C2O)C(O)C(O)C1O)C=CC=C(/C)C(=O)OC3OC(COC4OC(CO)C(O)C(O)C4O)C(O)C(O)C3O SEBIKDIMAPSUBY-JAUCNNNOSA-N 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 2
- 244000111489 Gardenia augusta Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 229930182470 glycoside Natural products 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 244000131522 Citrus pyriformis Species 0.000 description 1
- QWVMSYBGKWZIIE-RDFNRINOSA-N Flavochrome Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C1OC2(C)CCCC(C)(C)C2=C1)C=CC=C(/C)C=CC3C(=CCCC3(C)C)C QWVMSYBGKWZIIE-RDFNRINOSA-N 0.000 description 1
- 240000001972 Gardenia jasminoides Species 0.000 description 1
- 208000032456 Hemorrhagic Shock Diseases 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 206010049771 Shock haemorrhagic Diseases 0.000 description 1
- 238000003916 acid precipitation Methods 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 235000021466 carotenoid Nutrition 0.000 description 1
- 150000001747 carotenoids Chemical class 0.000 description 1
- 230000002279 cholagogic effect Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003810 ethyl acetate extraction Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- QWVMSYBGKWZIIE-FZKBJVJCSA-N flavochrome Chemical compound O1C2(C)CCCC(C)(C)C2=CC1C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1C(C)=CCCC1(C)C QWVMSYBGKWZIIE-FZKBJVJCSA-N 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Natural products CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 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
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- 238000004062 sedimentation Methods 0.000 description 1
- 239000011343 solid material Substances 0.000 description 1
Landscapes
- Pyridine Compounds (AREA)
Abstract
The invention discloses a kind of preparation method of high-purity trans-crocetin, belong to Chinese herbal medicine extracting separation field.The concrete steps of the method are: the trans-crocetin in crocetin crude product is changed into trans-crocetin pyridinium salt, through pyridine or the solution crystallization and the recrystallization that contain pyridine, obtain trans-crocetin pyridinium salt; High-purity trans-crocetin is obtained by after trans-crocetin pyridinium salt and acid-respons.The inventive method, have the advantage that operation steps is easy to repetition, operational condition is easy to control, recrystallisation solvent is easy to reclaim, be easy to industrial mass amplification, finished product trans-crocetin purity Absorbable organic halogens reaches more than 98%.
Description
Technical field
The invention belongs to Chinese herbal medicine extracting separation technology field, be specifically related to by the method for separating high-purity trans-crocetin further in crocetin crude product.
Background technology
Crocetin is the natural carotenoid constituents being present in the plant such as style and stigma of Saffron Crocus and cape jasmine, there is how unsaturated conjugated olefin(e) acid structure, be divided into trans-crocetin and 13-cis crocetin two kinds, except small part is free state, be present in the plant such as style and stigma of Saffron Crocus and cape jasmine mainly with being combined into ester glycosides (crocin and Gardenia Yellow) form with sugar.Trans-crocetin has pharmacologically active widely, comprises antitumor, atherosclerosis, hypertension and treatment hemorrhagic shock, also has the effects such as anti-oxidant and hepatic cholagogic.The structural formula of trans-crocetin is as follows:
Crocetin higher in conjunction with ester glycosides content in style and stigma of Saffron Crocus, but style and stigma of Saffron Crocus is expensive, therefrom extracts crocetin cost very high.Cape jasmine raw material sources are very abundant, and the Gardenia Yellow master therefrom extracted is containing crocin composition.Gardenia Yellow is the edible flavochrome of widespread use, and its production firm is numerous, commercially available product look valency from 50-500 not etc., by its under basic or acidic conditions according to a conventional method hydrolysis be easy to obtain crocetin crude product, existing commercially available product.But due to the insoluble of crocetin, to its separation and purification, especially the separating difficulty of trans-crocetin and 13-cis crocetin isomer is very large.Adopt conventional chromatography separating method, applied sample amount is all very little, and the eluting solvent of meeting at substantial, be not suitable for its mass-producing preparation.And when adopting solvent crystallization method, because it is insoluble in most organic solvent, be difficult to find desirable recrystallisation solvent.Such as, there is investigator that crocetin crude product Acid precipitation method is refined trans-crocetin, its precipitation process is noncrystalline process, cis crocetin and other impurity can along with sedimentations together, the condition such as pH value, sample concentration, settling velocity, time of repose that gained trans-crocetin purity and acid are sunk in process is closely related, condition is wayward, poor reproducibility, is unfavorable for industrialized mass production.Investigator is also had to adopt N, dinethylformamide and methyl-sulphoxide crystallization purifying crocetin.But the boiling point of N, dinethylformamide and methyl-sulphoxide crystallization is higher, and easily destroy sample structure in removal process, finished product may have higher dissolvent residual.
Summary of the invention
The object of this invention is to provide the method preparing trans-crocetin from crocetin crude product, the method treatment condition are easy to control, and recrystallisation solvent is easy to reclaim, and the trans-crocetin purity obtained is high, is beneficial to suitability for industrialized production.
The present invention is achieved through the following technical solutions:
The preparation method of high-purity trans-crocetin, changes into trans-crocetin pyridinium salt by the trans-crocetin in crocetin crude product, through pyridine or the solution crystallization and the recrystallization that contain pyridine, obtains trans-crocetin pyridinium salt; High-purity trans-crocetin is obtained by after trans-crocetin pyridinium salt and acid-respons.
In described crocetin crude product, the mass percent of trans-crocetin is 5% ~ 95%.
The described solution be made up of mixture and the pyridine of one or two or more kinds in water, ethanol, methyl alcohol, acetone, n-propyl alcohol, Virahol, propyl carbinol, isopropylcarbinol, tetrahydrofuran (THF) and acetonitrile containing the solution of pyridine; Described is 50% ~ 100% containing the mass percent of pyridine in the solution of pyridine.
During described crystallization, pyridine or the consumption containing the solution of pyridine are 30 ~ 500 times of crocetin crude product weight, and during recrystallization, pyridine or the solution usage containing pyridine are 50 ~ 1000 times of trans-crocetin pyridinium salt weight, and the number of times of recrystallization is 1-4 time.
When described crystallization and recrystallization, temperature is-20 ~ 25 DEG C, and the crystallization time is 0.5 ~ 48 hour.
After described trans-crocetin pyridinium salt and acid-respons the concrete grammar of high-purity trans-crocetin adopts one in following method:
A. the mixed solvent of trans-crocetin pyridinium salt alkaline aqueous solution or alkaline aqueous solution and hydrophilic organic solvent is dissolved, then with acid for adjusting pH value to 1 ~ 6, separate out precipitation, after getting precipitation washing, drying, namely obtain high-purity trans-crocetin;
B. by trans-crocetin pyridinium salt hydrophilic organic solvent or its aqueous dissolution, then with acid for adjusting pH value to 1 ~ 6, separate out precipitation, after getting precipitation washing, drying, namely obtain high-purity trans-crocetin;
C. trans-crocetin pyridinium salt being suspended in pH value is in the acidic aqueous solution of 1 ~ 6, then acidic aqueous solution layer is extracted to hydrophobic organic solvent closely colourless, merge hydrophobic organic solvent layer, be washed to elutant in neutral, reclaim organic solvent, drying, obtains high-purity trans-crocetin;
In described step a, b, hydrophilic organic solvent is selected from one or more the mixed solvent in ethanol, methyl alcohol, acetone, n-propyl alcohol, Virahol, tetrahydrofuran (THF), acetonitrile; In described step c, hydrophobic organic solvent is one or more the mixed solvent in vinyl acetic monomer, propyl carbinol, isopropylcarbinol.
The described step a neutral and alkali aqueous solution is the aqueous solution of sodium hydroxide, potassium hydroxide, sodium carbonate, salt of wormwood, sodium bicarbonate or saleratus, or ammoniacal liquor; In described alkaline aqueous solution, the mass percentage concentration of alkali is 0.1% ~ 5%.
In described step a, b, adjust ph acid used adopts hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid or citric acid; In step c, acidic aqueous solution used is hydrochloric acid or sulfuric acid, phosphoric acid, acetic acid or lemon aqueous acid.
the present invention has following beneficial effect:
1, trans-crocetin is insoluble in the common solvent such as water, ethanol, methyl alcohol, acetone, n-propyl alcohol, Virahol, propyl carbinol, isopropylcarbinol, tetrahydrofuran (THF) and acetonitrile, is comparatively soluble in pyridine and basic solvent.The present invention is first by the trans-crocetin in crocetin crude product and pyridine salify, and it is easy to crystallization, thus is separated smoothly with other impurity.Trans-crocetin pyridinium salt is through recrystallization purifying, and purity can reach more than 98%, then itself and acid-respons are obtained trans-crocetin, simple to operate, and in finished product, trans-crocetin purity Absorbable organic halogens reaches more than 98%.In crocetin crude product, trans-crocetin content is higher, and the required placement crystallization time is shorter.Trans-crocetin pyridinium salt is compared with trans-crocetin, comparatively be soluble in hydrophilic solvent and the water-containing solvents thereof such as ethanol, methyl alcohol, acetone, n-propyl alcohol, Virahol, propyl carbinol, isopropylcarbinol, tetrahydrofuran (THF) and acetonitrile, if basic solvent, solubleness is larger, trans-crocetin can be reverted to easily after being acidified with acid, solubleness declines, and separates out, reach comparatively ideal purifying object with precipitation forms.
2, recrystallisation solvent boiling point used of the present invention is lower, and it is convenient to reclaim, and repeatedly can recycle, have fewer environmental impacts.
3, production technique of the present invention is simple, and condition is easy to control, and can realize more easily from being prepared into production in enormous quantities in a small amount.
Embodiment
The present invention is further illustrated to be below to provide most preferred embodiment, but and unrestricted the present invention.
The trans-crocetin prepared in the embodiment of the present invention and pyridinium salt thereof, adopt high performance liquid chromatograph detection level.Actual conditions is: Shimadzu high performance liquid chromatograph (LC-20AD double pump and Sil-20A automatic sample handling system, SPD-20A detector, LC solution chromatographic working station), determined wavelength: 423nm; C
18-ODS post; Moving phase is methanol-water-Glacial acetic acid (79.5:20:0.5); Flow velocity: 1.0ml/min; Sample feeding concentration about 10 μ g/ml, sample size is 20 μ l.
embodiment 1
The preparation of crocetin crude product: get the Gardenia Yellow 500g that commercially available look valency is 60, add 2% aqueous sodium hydroxide solution 10L to dissolve, in 60 DEG C of hydrolysis 2.5 hours, add the salt acid for adjusting pH value to 3.0 that mass percentage concentration is 10%, leave standstill 24 hours and separate out precipitation, centrifugal, get precipitation and wash with water to washing out water liquid pH value in neutral.Gained is deposited in 60 DEG C of vacuum-dryings 3 hours, obtains crocetin crude product 20.5g, detecting wherein trans-crocetin mass percent through HPLC is 14.6%.
Get crocetin crude product 5.0g prepared by the present embodiment, add 300mL pyridine, be heated to 80 DEG C and make it dissolve, insoluble impurity filtered while hot removing.Filtrate is placed 24 hours in room temperature (20-25 DEG C), separates out trans-crocetin pyridinium salt crystal.By trans-crocetin pyridinium salt crystal pyridine recrystallization 3 times (each solvent load is 200 times of crystal weight), obtain highly purified trans-crocetin pyridinium salt, detecting its purity through HPLC is 99.3%.The aqueous sodium hydroxide solution being 1% by highly purified trans-crocetin pyridinium salt mass percentage concentration dissolves completely, add concentrated hydrochloric acid adjust ph to 3.0, leave standstill 24 hours and separate out trans-crocetin precipitation, centrifugal, get precipitation water and be repeatedly washed till elutant pH value in neutral.By washing after be deposited in 60 DEG C of vacuum-dryings 3 hours, obtain trans-crocetin 0.21g, detecting its purity through HPLC is 99.2%.
embodiment 2
The crocetin crude product 5.0g of preparation in Example 1, adds the pyridine solution that 300mL mass percentage concentration is 80%, is heated to 70 DEG C of dissolvings, insoluble impurity filtered while hot removing.Filtrate is placed 24 hours in refrigerator cold-storage layer (2-8 DEG C), separate out trans-crocetin pyridinium salt crystal. be the pyridine solution recrystallization 3 times (each solvent load is 250 times of crystal) of 80% by trans-crocetin pyridinium salt crystal mass percentage concentration, obtain highly purified trans-crocetin pyridinium salt, detecting its purity through HPLC is 99.4%.The aqueous ethanolic solution that concentration expressed in percentage by volume is 95% is added in highly purified trans-crocetin pyridinium salt, be heated to dissolve completely, with the salt acid for adjusting pH value to 2.0 that mass percentage concentration is 10%, leave standstill 24 hours and separate out trans-crocetin precipitation, centrifugal, get precipitation water and be repeatedly washed till and wash out water liquid pH value in neutral.By washing after be deposited in 60 DEG C of vacuum-dryings 3 hours, obtain trans-crocetin 0.15g, detecting its purity through HPLC is 99.5%.
embodiment 3
The crocetin crude product 5.0g of preparation in Example 1, adds the pyridine solution (pyridine quality hundred number is 60%) of 300mL ethanol, is heated to 80 DEG C and makes dissolution of solid, insoluble impurity filtered while hot removing.Filtrate is placed 24 hours in refrigerator cold-storage layer (2-8 DEG C), separates out trans-crocetin pyridinium salt crystal.By pyridine solution (pyridine quality hundred number the is 60%) recrystallization 3 times (each solvent load is 300 times of crystal weight) of trans-crocetin pyridinium salt crystal ethanol, obtain highly purified trans-crocetin pyridinium salt, detecting its purity through HPLC is 98.8%.It is in the aqueous hydrochloric acid of 2% that highly purified trans-crocetin pyridinium salt is suspended in 1000ml mass percentage concentration, then closely colourless to acidic aqueous solution layer with ethyl acetate extraction, merge vinyl acetic monomer layer, it is neutral for washing with water to elutant, obtains trans-crocetin after recovery of acetic acid ethyl ester.By trans-crocetin in 60 DEG C of vacuum-dryings 2 hours, obtain 0.20g trans-crocetin, detecting its purity through HPLC is 98.6%.
embodiment 4
The preparation of crocetin crude product: get the Gardenia Yellow 500g that commercially available look valency is 500, add 2% aqueous sodium hydroxide solution 10L to dissolve, in 60 DEG C of hydrolysis 3 hours, adding 10L concentration expressed in percentage by volume is 95% ethanol, and mixing, adds the sulfuric acid adjust ph to 2.0 that mass percentage concentration is 5%, leave standstill 24 hours and separate out precipitation, filtering, get precipitation, washing with water to washing out water liquid pH value in neutral.Gained is deposited in 60 DEG C of vacuum-dryings 3 hours, obtains crocetin crude product 45.9g(crocetin crude product I), detecting wherein trans-crocetin content through HPLC is 44.6%.Get 20.0g crocetin crude product I, add the methanol solution that mass percentage concentration is the potassium hydroxide of 1%, heating makes to dissolve completely, add the salt acid for adjusting pH value to 5.0 that mass percentage concentration is 5%, leave standstill 24 hours and separate out precipitation, centrifugal, get precipitation and wash with water to washing out water liquid pH value in neutral.Gained is deposited in 60 DEG C of vacuum-dryings 3 hours, obtains crocetin crude product 9.2g(crocetin crude product II), detect through HPLC, wherein trans-crocetin mass percent is 75.3%.
Get crocetin crude product I prepared by 5.0g the present embodiment, add 500mL pyridine, be heated to 80 DEG C and make dissolution of solid, insoluble impurity filtered while hot removing.Filtrate is placed 2 hours in-20 DEG C, separates out trans-crocetin pyridinium salt crystal.By trans-crocetin pyridinium salt crystal pyridine recrystallization 2 times (each solvent load is 200 times of crystal weight), obtain highly purified trans-crocetin pyridinium salt, detecting its purity through HPLC is 99.5%.The potassium hydroxide aqueous solution being 2% by highly purified trans-crocetin pyridinium salt mass percentage concentration dissolves completely, by the phosphate aqueous solution adjust ph to 2.0 that mass concentration is 5%, leave standstill 24 hours and separate out trans-crocetin precipitation, centrifugal, get precipitation water and be repeatedly washed till elutant pH value in neutral.Gained is deposited in 60 DEG C of vacuum-dryings 2 hours, obtains trans-crocetin 0.88g, detecting its purity through HPLC is 99.5%.
embodiment 5
Get the crocetin crude product I of preparation in 5.0g embodiment 4, add the pyridine solution (pyridine quality hundred number is 70%) of 600mL acetone, be heated to 40 DEG C and make solid material dissolves, insoluble impurity filtered while hot removing.Filtrate is placed 24 hours in refrigerator cold-storage layer (2-8 DEG C), separates out trans-crocetin pyridinium salt crystal.By pyridine solution (pyridine quality hundred number the is 70%) recrystallization 2 times (each solvent load is 300 times of crystal weight) of trans-crocetin pyridinium salt crystal acetone, obtain highly purified trans-crocetin pyridinium salt, detecting its purity through HPLC is 99.6%.It is in the aqueous sulfuric acid of 2% that highly purified trans-crocetin pyridinium salt is suspended in 1000ml concentration expressed in percentage by weight, then closely colourless to aqueous sulfuric acid layer with n-butanol extraction, merge n-butanol layer, be washed to and wash out water liquid in neutral, after reclaiming propyl carbinol, obtain trans-crocetin.By trans-crocetin in 60 DEG C of vacuum-dryings 4 hours, obtain trans-crocetin 0.94g, detecting its purity through HPLC is 99.4%.
embodiment 6
Get the crocetin crude product I of preparation in 3.0g embodiment 4, add the pyridine solution (pyridine quality hundred number is 90%) of 200mL Virahol, be heated to 70 DEG C and make dissolution of solid, insoluble impurity filtered while hot removing.Filtrate is placed 2 hours in-20 DEG C, separates out trans-crocetin pyridinium salt crystal.By pyridine solution (pyridine quality hundred number the be 90%) recrystallization 1 time of trans-crocetin pyridinium salt crystal with the Virahol of its weight 200 times, obtain highly purified trans-crocetin pyridinium salt, detecting its purity through HPLC is 98.4%.It is in the methanol solution of the potassium hydroxide of 1% that highly purified trans-crocetin pyridinium salt is added mass percentage concentration, heating makes to dissolve completely, by the aqueous hydrochloric acid adjust ph to 2.0 that mass percentage concentration is 5%, leave standstill 24 hours and separate out precipitation, filter, get precipitation and wash pH value to elutant with water in neutral.By washing after be deposited in 60 DEG C of vacuum-dryings 2 hours, obtain trans-crocetin 0.43g, detecting its purity through HPLC is 98.5%.
embodiment 7
Get the crocetin crude product I of preparation in 3.0g embodiment 4, add the pyridine solution (pyridine quality hundred number is 85%) of 200mL propyl carbinol, be heated to 70 DEG C and make dissolution of solid, insoluble impurity filtered while hot removing.Filtrate is placed 2 hours in-20 DEG C, separates out trans-crocetin pyridinium salt crystal.By pyridine solution (pyridine quality hundred number the is 85%) recrystallization 2 times (each solvent load is 200 times of crystal weight) of trans-crocetin pyridinium salt crystal propyl carbinol, obtain highly purified trans-crocetin pyridinium salt, detecting its purity through HPLC is 99.6%.In highly purified trans-crocetin pyridinium salt, add the potassium hydroxide aqueous solution that mass percentage concentration is 2%, be heated to dissolve completely, by Glacial acetic acid adjust ph to 4.0, leave standstill 24 hours and separate out precipitation, filter, get precipitation, wash pH value to elutant with water in neutral.By washing after be deposited in 60 DEG C of vacuum-dryings 2 hours, obtain trans-crocetin 0.51g, detecting its purity through HPLC is 99.6%.
embodiment 8
Get the crocetin crude product I of preparation in 3.0g embodiment 4, add the pyridine solution (pyridine quality hundred number is 55%) of 250mL acetonitrile, be heated to 80 DEG C of dissolvings, insoluble impurity filtered while hot removing.Filtrate is placed 12 hours in refrigerator cold-storage layer (2-8 DEG C), separates out trans-crocetin pyridinium salt crystal.By pyridine solution (pyridine quality hundred number the is 55%) recrystallization 1 time (solvent load is 300 times of crystal weight) of trans-crocetin pyridinium salt crystal acetonitrile, obtain highly purified trans-crocetin pyridinium salt, detecting its purity through HPLC is 99.1%.Be the mass mixing such as aqueous sodium hydroxide solution and 95% ethanol of 1% by mass percentage concentration, add highly purified trans-crocetin pyridinium salt wherein, be heated to dissolve completely.By the aqueous hydrochloric acid adjust ph to 4.0 that mass percentage concentration is 5%, leave standstill 24 hours and separate out precipitation, filter, get precipitation and wash pH value to elutant with water in neutral.By washing after be deposited in 60 DEG C of vacuum-dryings 2 hours, obtain trans-crocetin 0.45g, detecting its purity through HPLC is 99.1%.
embodiment 9
Get the crocetin crude product II of preparation in 3.0g embodiment 4, add the pyridine solution (pyridine quality hundred number is 75%) of 250mL tetrahydrofuran (THF), be heated to 80 DEG C and make dissolution of solid, insoluble impurity filtered while hot removing.Filtrate is placed 12 hours in refrigerator cold-storage layer (2-8 DEG C), separates out trans-crocetin pyridinium salt crystal.By pyridine solution (pyridine quality hundred number the is 75%) recrystallization 2 times (each solvent load is 300 times of crystal weight) of trans-crocetin pyridinium salt crystal tetrahydrofuran (THF), obtain highly purified trans-crocetin pyridinium salt, detecting its purity through HPLC is 99.5%.In highly purified trans-crocetin pyridinium salt, add Virahol, heating makes to dissolve completely, is the aqueous sulfuric acid adjust ph to 2.0 of 5% by mass percentage concentration, leaves standstill 24 hours and separates out precipitation, filters, and gets precipitation and washes with water to elutant pH value in neutral.By washing after be deposited in 60 DEG C of vacuum-dryings 2 hours, obtain trans-crocetin 1.05g, detecting its purity through HPLC is 99.6%.
embodiment 10
Get commercially available crocetin crude product (trans-crocetin mass percent is 73.9%) 5.0g, add 400mL pyridine, be heated to 80 DEG C and make dissolution of solid, insoluble impurity filtered while hot removing.Filtrate is placed 24 hours in room temperature (20-25 DEG C), separates out trans-crocetin pyridinium salt crystal.By the pyridine recrystallization 1 time of trans-crocetin pyridinium salt crystal by its weight 200 times, obtain highly purified trans-crocetin pyridinium salt, detecting its purity through HPLC is 99.4%.The ammonia aqueous solution being 1% by highly purified trans-crocetin pyridinium salt mass percentage concentration makes it dissolve completely, with the salt acid for adjusting pH value to 3.0 that mass percentage concentration is 5%, leave standstill 24 hours and separate out precipitation, filter, get precipitation and wash with water to washing out water liquid pH value in neutral.By washing after be deposited in 60 DEG C of vacuum-dryings 2 hours, obtain trans-crocetin 2.06g, detecting its purity through HPLC is 99.3%.
Claims (6)
1. the preparation method of trans-crocetin, is characterized in that: the trans-crocetin in crocetin crude product is changed into trans-crocetin pyridinium salt, through pyridine or the solution crystallization and the recrystallization that contain pyridine, obtains trans-crocetin pyridinium salt; High-purity trans-crocetin is obtained by after trans-crocetin pyridinium salt and acid-respons; The described solution be made up of mixture and the pyridine of one or two or more kinds in water, ethanol, methyl alcohol, acetone, n-propyl alcohol, Virahol, propyl carbinol, isopropylcarbinol, tetrahydrofuran (THF) and acetonitrile containing the solution of pyridine; Described is 50 ~ 100% containing the mass percent of pyridine in the solution of pyridine; During described crystallization, pyridine or the consumption containing the solution of pyridine are 30 ~ 500 times of crocetin crude product weight, and during recrystallization, pyridine or the solution usage containing pyridine are 50 ~ 1000 times of trans-crocetin pyridinium salt weight, and the number of times of recrystallization is 1-4 time.
2. the preparation method of trans-crocetin according to claim 1, is characterized in that: in described crocetin crude product, the mass percent of trans-crocetin is 5% ~ 95%.
3. the preparation method of trans-crocetin according to claim 2, it is characterized in that: when described crystallization and recrystallization, temperature is-20 ~ 25 DEG C, the crystallization time is 0.5 ~ 48 hour.
4. the preparation method of trans-crocetin according to claim 3, is characterized in that: after described trans-crocetin pyridinium salt and acid-respons the concrete grammar of trans-crocetin adopts one in following method:
A. the mixed solvent of trans-crocetin pyridinium salt alkaline aqueous solution or alkaline aqueous solution and hydrophilic organic solvent is dissolved, then with acid for adjusting pH value to 1 ~ 6, separate out precipitation, after getting precipitation washing, drying, namely obtain trans-crocetin;
B. by trans-crocetin pyridinium salt hydrophilic organic solvent or its aqueous dissolution, then with acid for adjusting pH value to 1 ~ 6, separate out precipitation, after getting precipitation washing, drying, namely obtain trans-crocetin;
C. trans-crocetin pyridinium salt being suspended in pH value is in the acidic aqueous solution of 1 ~ 6, is then extracted to acidic aqueous solution layer with hydrophobic organic solvent closely colourless, merges hydrophobic organic solvent layer, be washed to elutant in neutral, reclaim organic solvent, dry, obtain trans-crocetin;
In described step a, b, hydrophilic organic solvent is selected from one or more the mixed solvent in ethanol, methyl alcohol, acetone, n-propyl alcohol, Virahol, tetrahydrofuran (THF), acetonitrile; In described step c, hydrophobic organic solvent is one or more the mixed solvent in vinyl acetic monomer, propyl carbinol, isopropylcarbinol.
5. the preparation method of trans-crocetin according to claim 4, is characterized in that the described step a neutral and alkali aqueous solution is the aqueous solution of sodium hydroxide, potassium hydroxide, sodium carbonate, salt of wormwood, sodium bicarbonate or saleratus, or ammoniacal liquor; In described alkaline aqueous solution, the mass percentage concentration of alkali is 0.1 ~ 5%.
6. the preparation method of trans-crocetin according to claim 4, is characterized in that in described step a, b, adjust ph acid used adopts hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid or citric acid; In step c, acidic aqueous solution used is hydrochloric acid or sulfuric acid, phosphoric acid, acetic acid or lemon aqueous acid.
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| CN108403677B (en) * | 2018-05-03 | 2020-06-05 | 北京市心肺血管疾病研究所 | Application of crocin in prevention and treatment of thoracic aortic dissection/aortic aneurysm |
| CN109180469B (en) | 2018-09-05 | 2021-04-13 | 嘉文丽（福建）化妆品有限公司 | Separation method of cis-trans isomeric crocetin |
| CN109620831A (en) * | 2018-12-20 | 2019-04-16 | 浙江工业大学 | Application of the crocetin in preparation treatment nonalcoholic steatohepatitis drug |
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| WO2004078695A1 (en) * | 2003-03-07 | 2004-09-16 | Riken Vitamin Co., Ltd. | Method for the purification of crocetin |
| CN101225040A (en) * | 2007-01-18 | 2008-07-23 | 中国科学院过程工程研究所 | Method for extracting crocetin from gardenia |
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| WO2004078695A1 (en) * | 2003-03-07 | 2004-09-16 | Riken Vitamin Co., Ltd. | Method for the purification of crocetin |
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