CN114317630A - Preparation method of diosmetin - Google Patents
Preparation method of diosmetin Download PDFInfo
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- CN114317630A CN114317630A CN202111255615.5A CN202111255615A CN114317630A CN 114317630 A CN114317630 A CN 114317630A CN 202111255615 A CN202111255615 A CN 202111255615A CN 114317630 A CN114317630 A CN 114317630A
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- diosmin
- enzymolysis
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- MBNGWHIJMBWFHU-UHFFFAOYSA-N diosmetin Chemical compound C1=C(O)C(OC)=CC=C1C1=CC(=O)C2=C(O)C=C(O)C=C2O1 MBNGWHIJMBWFHU-UHFFFAOYSA-N 0.000 title claims abstract description 37
- QAGGICSUEVNSGH-UHFFFAOYSA-N Diosmetin Natural products C1=C(O)C(OC)=CC=C1C1=CC(=O)C2=CC=C(O)C=C2O1 QAGGICSUEVNSGH-UHFFFAOYSA-N 0.000 title claims abstract description 36
- 229960001876 diosmetin Drugs 0.000 title claims abstract description 35
- 235000015428 diosmetin Nutrition 0.000 title claims abstract description 35
- 238000002360 preparation method Methods 0.000 title claims abstract description 30
- VUYDGVRIQRPHFX-UHFFFAOYSA-N hesperidin Natural products COc1cc(ccc1O)C2CC(=O)c3c(O)cc(OC4OC(COC5OC(O)C(O)C(O)C5O)C(O)C(O)C4O)cc3O2 VUYDGVRIQRPHFX-UHFFFAOYSA-N 0.000 claims abstract description 85
- GZSOSUNBTXMUFQ-NJGQXECBSA-N 5,7,3'-Trihydroxy-4'-methoxyflavone 7-O-rutinoside Natural products O(C[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](Oc2cc(O)c3C(=O)C=C(c4cc(O)c(OC)cc4)Oc3c2)O1)[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](C)O1 GZSOSUNBTXMUFQ-NJGQXECBSA-N 0.000 claims abstract description 51
- GZSOSUNBTXMUFQ-YFAPSIMESA-N diosmin Chemical compound C1=C(O)C(OC)=CC=C1C(OC1=C2)=CC(=O)C1=C(O)C=C2O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@H]2[C@@H]([C@H](O)[C@@H](O)[C@H](C)O2)O)O1 GZSOSUNBTXMUFQ-YFAPSIMESA-N 0.000 claims abstract description 51
- 229960004352 diosmin Drugs 0.000 claims abstract description 51
- IGBKNLGEMMEWKD-UHFFFAOYSA-N diosmin Natural products COc1ccc(cc1)C2=C(O)C(=O)c3c(O)cc(OC4OC(COC5OC(C)C(O)C(O)C5O)C(O)C(O)C4O)cc3O2 IGBKNLGEMMEWKD-UHFFFAOYSA-N 0.000 claims abstract description 51
- 108090000790 Enzymes Proteins 0.000 claims abstract description 29
- 102000004190 Enzymes Human genes 0.000 claims abstract description 29
- 238000006243 chemical reaction Methods 0.000 claims abstract description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 47
- 238000001914 filtration Methods 0.000 claims description 39
- 239000001100 (2S)-5,7-dihydroxy-2-(3-hydroxy-4-methoxyphenyl)chroman-4-one Substances 0.000 claims description 34
- QUQPHWDTPGMPEX-UHFFFAOYSA-N Hesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(COC4C(C(O)C(O)C(C)O4)O)O3)O)=CC(O)=C2C(=O)C1 QUQPHWDTPGMPEX-UHFFFAOYSA-N 0.000 claims description 34
- QUQPHWDTPGMPEX-UTWYECKDSA-N aurantiamarin Natural products COc1ccc(cc1O)[C@H]1CC(=O)c2c(O)cc(O[C@@H]3O[C@H](CO[C@@H]4O[C@@H](C)[C@H](O)[C@@H](O)[C@H]4O)[C@@H](O)[C@H](O)[C@H]3O)cc2O1 QUQPHWDTPGMPEX-UTWYECKDSA-N 0.000 claims description 34
- APSNPMVGBGZYAJ-GLOOOPAXSA-N clematine Natural products COc1cc(ccc1O)[C@@H]2CC(=O)c3c(O)cc(O[C@@H]4O[C@H](CO[C@H]5O[C@@H](C)[C@H](O)[C@@H](O)[C@H]5O)[C@@H](O)[C@H](O)[C@H]4O)cc3O2 APSNPMVGBGZYAJ-GLOOOPAXSA-N 0.000 claims description 34
- QUQPHWDTPGMPEX-QJBIFVCTSA-N hesperidin Chemical compound C1=C(O)C(OC)=CC=C1[C@H]1OC2=CC(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO[C@H]4[C@@H]([C@H](O)[C@@H](O)[C@H](C)O4)O)O3)O)=CC(O)=C2C(=O)C1 QUQPHWDTPGMPEX-QJBIFVCTSA-N 0.000 claims description 34
- 229940025878 hesperidin Drugs 0.000 claims description 34
- ARGKVCXINMKCAZ-UHFFFAOYSA-N neohesperidine Natural products C1=C(O)C(OC)=CC=C1C1OC2=CC(OC3C(C(O)C(O)C(CO)O3)OC3C(C(O)C(O)C(C)O3)O)=CC(O)=C2C(=O)C1 ARGKVCXINMKCAZ-UHFFFAOYSA-N 0.000 claims description 34
- 239000007810 chemical reaction solvent Substances 0.000 claims description 31
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 30
- 239000000243 solution Substances 0.000 claims description 29
- 238000002156 mixing Methods 0.000 claims description 28
- 239000002994 raw material Substances 0.000 claims description 27
- 239000012065 filter cake Substances 0.000 claims description 22
- 238000005406 washing Methods 0.000 claims description 18
- 239000000706 filtrate Substances 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 15
- 238000010438 heat treatment Methods 0.000 claims description 14
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 12
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 claims description 10
- 108010030923 hesperidinase Proteins 0.000 claims description 10
- 239000011630 iodine Substances 0.000 claims description 10
- 229910052740 iodine Inorganic materials 0.000 claims description 10
- 238000004519 manufacturing process Methods 0.000 claims description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims description 5
- 108010056771 Glucosidases Proteins 0.000 claims description 4
- 102000004366 Glucosidases Human genes 0.000 claims description 4
- 108010059820 Polygalacturonase Proteins 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 108010093305 exopolygalacturonase Proteins 0.000 claims description 4
- 108010001078 naringinase Proteins 0.000 claims description 4
- 238000010298 pulverizing process Methods 0.000 claims description 4
- 239000012670 alkaline solution Substances 0.000 claims description 3
- PNDPGZBMCMUPRI-UHFFFAOYSA-N iodine Chemical compound II PNDPGZBMCMUPRI-UHFFFAOYSA-N 0.000 claims description 3
- -1 rhamnosidase Proteins 0.000 claims description 3
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 10
- 239000002351 wastewater Substances 0.000 abstract description 4
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 30
- 238000010992 reflux Methods 0.000 description 13
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 12
- 238000003756 stirring Methods 0.000 description 9
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 8
- 150000001408 amides Chemical class 0.000 description 8
- 239000003795 chemical substances by application Substances 0.000 description 8
- 230000007071 enzymatic hydrolysis Effects 0.000 description 8
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 8
- 238000000605 extraction Methods 0.000 description 8
- 239000002904 solvent Substances 0.000 description 8
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 description 6
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical group C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 6
- 239000000498 cooling water Substances 0.000 description 5
- 238000002425 crystallisation Methods 0.000 description 5
- 230000008025 crystallization Effects 0.000 description 5
- 238000010812 external standard method Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000002481 ethanol extraction Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 229930003935 flavonoid Natural products 0.000 description 3
- 150000002215 flavonoids Chemical class 0.000 description 3
- 235000017173 flavonoids Nutrition 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000000643 oven drying Methods 0.000 description 3
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 3
- 239000013558 reference substance Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- FXHOOIRPVKKKFG-UHFFFAOYSA-N N,N-Dimethylacetamide Chemical compound CN(C)C(C)=O FXHOOIRPVKKKFG-UHFFFAOYSA-N 0.000 description 2
- PMDCZENCAXMSOU-UHFFFAOYSA-N N-ethylacetamide Chemical compound CCNC(C)=O PMDCZENCAXMSOU-UHFFFAOYSA-N 0.000 description 2
- OHLUUHNLEMFGTQ-UHFFFAOYSA-N N-methylacetamide Chemical compound CNC(C)=O OHLUUHNLEMFGTQ-UHFFFAOYSA-N 0.000 description 2
- 230000006978 adaptation Effects 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 235000005911 diet Nutrition 0.000 description 2
- 230000037213 diet Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000003912 environmental pollution Methods 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- IPZCJUOJSODZNK-UHFFFAOYSA-N n,n'-dimethyloxamide Chemical compound CNC(=O)C(=O)NC IPZCJUOJSODZNK-UHFFFAOYSA-N 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 description 2
- 239000012088 reference solution Substances 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 239000012085 test solution Substances 0.000 description 2
- 238000009210 therapy by ultrasound Methods 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 235000017060 Arachis glabrata Nutrition 0.000 description 1
- 244000105624 Arachis hypogaea Species 0.000 description 1
- 235000010777 Arachis hypogaea Nutrition 0.000 description 1
- 235000018262 Arachis monticola Nutrition 0.000 description 1
- 235000007516 Chrysanthemum Nutrition 0.000 description 1
- 244000189548 Chrysanthemum x morifolium Species 0.000 description 1
- 235000005979 Citrus limon Nutrition 0.000 description 1
- 244000131522 Citrus pyriformis Species 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- 244000024873 Mentha crispa Species 0.000 description 1
- 235000014749 Mentha crispa Nutrition 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241000606265 Valeriana jatamansi Species 0.000 description 1
- AOZUYISQWWJMJC-UHFFFAOYSA-N acetic acid;methanol;hydrate Chemical compound O.OC.CC(O)=O AOZUYISQWWJMJC-UHFFFAOYSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000002113 chemopreventative effect Effects 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 150000008131 glucosides Chemical class 0.000 description 1
- 125000003147 glycosyl group Chemical group 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 235000020232 peanut Nutrition 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a preparation method of diosmetin, which comprises the following steps: diosmin is subjected to enzymolysis by using enzyme; the weight ratio of the diosmin to the enzyme is 100 (0.60-23.81); the preparation method does not produce a large amount of waste water, does not pollute the environment, is easy to control reaction conditions, simple to operate, convenient to treat and high in yield, and the prepared diosmetin has high HPLC content, high purity and good product quality.
Description
Technical Field
The invention relates to the technical field of synthesis, and particularly relates to a preparation method of diosmetin.
Background
Diosmetin is one of natural flavonoids, and has a chemical name of 3 ', 5, 7-trihydroxy-4' -methoxyflavone. The diosmetin is yellow powder, has a melting point of 256-258 ℃, a CAS number of 520-34-3, a molecular formula of C16H12O6, a molecular weight of 300.26, and can be dissolved in organic solvents such as methanol, ethanol and the like. The hydroxyl groups at three positions of 3', 5 and 7 and the C2 ═ C3 double bond in the structural formula determine the unique chemical properties and biological activities of diosmetin, wherein the phenolic hydroxyl group at the position 7 can be combined with different glycosyl groups to generate glycosylation reaction to show different biological activities. Diosmetin is widely distributed in nature, mainly exists in a free type or a glucoside type, and is mainly found in natural medicines such as chrysanthemum, spearmint, valeriana jatamansi jones and fruits such as lemon, peanut and the like at present. The flavonoids are important components of daily diet of people, the chemoprevention effect of the flavonoids in diet on cancer is a hot point of study of researchers in recent years, and diosmetin has the effects of oxidation resistance, infection resistance, shock resistance and the like, can be widely applied to functional foods, cosmetics and medicines, and therefore has received wide attention.
The content of diosmetin in natural plants is low, so that a synthesis method is mostly adopted to prepare diosmetin to meet market demands, and the existing diosmetin synthesis process adopts organic solvents and inorganic acids as hydrolysis media, so that a large amount of acidic wastewater is generated, and environmental pollution is easily caused.
Disclosure of Invention
In view of the above, the application provides a method for preparing diosmin, which prepares diosmin by enzymolysis, and does not generate a large amount of wastewater in the production process, and does not cause environmental pollution, the reaction conditions are easy to control, the operation is simple, the treatment is convenient, the yield is high, the prepared diosmin has high HPLC content, the HPLC content reaches more than 98%, the purity is high, and the product quality reaches 99%.
In order to solve the technical problems, the technical scheme provided by the application is a preparation method of diosmetin, which comprises the following steps: diosmin is subjected to enzymolysis by using enzyme; the weight ratio of diosmin to enzyme is 100 (0.60-23.81).
Preferably, the preparation method specifically comprises: mixing diosmin, enzyme and water for enzymolysis; the weight ratio of the diosmin to the enzyme is 100 (0.60-23.81); the ratio of diosmin to water was 105 kg: 1m3。
Preferably, the enzyme is selected from one or more of hesperidinase, rhamnosidase, naringinase, glucosidase and pectinase.
Preferably, the ratio of diosmin to enzyme by weight is 100: 2.38.
Preferably, the enzyme is hesperidinase.
Preferably, the enzymolysis temperature is 35-70 ℃, and the enzymolysis time is 12-72 h.
Preferably, the enzymolysis temperature is 40-50 ℃.
Preferably, the enzymatic hydrolysis temperature is 50 ℃.
Preferably, the enzymolysis time is 24 h.
Preferably, the preparation method further comprises: mixing the reaction solvent, hesperidin, alkaline reagent and iodine, and heating for reaction to obtain diosmin.
Preferably, the preparation method further comprises:
(A1) mixing iodine and a reaction solvent to obtain a raw material solution I;
(A2) mixing hesperidin with a reaction solvent to obtain a raw material solution II;
(A3) mixing the raw material solution I and the raw material solution II, heating for reaction, recovering a reaction solvent, adding water, uniformly mixing, filtering, washing with water, and collecting a filter cake;
(A4) mixing and dissolving the filter cake and an alkaline solution, and filtering by using a fine filter to obtain a fine-filtered filtrate;
(A5) and mixing the fine filtered filtrate with hydrochloric acid, crystallizing, filtering, and collecting a filter cake to obtain diosmin.
Preferably, the weight ratio of the hesperidin to the enzyme is 100 (0.5-20).
Preferably, the hesperidin to enzyme weight ratio is 100:2.
Preferably, the weight ratio of iodine to hesperidin is 1: 5 to 15.
Preferably, the weight ratio of iodine to hesperidin is 3: 10.
preferably, the ratio of the reaction solvent to the iodine used in the step (A1) is 1m3：75kg
Preferably, the ratio of the reaction solvent to the hesperidin used in the step (A2) is 1m3：200～300kg。
Preferably, the ratio of the reaction solvent to the hesperidin used in the step (A2) is 1m3：250kg。
Preferably, the step (a3) specifically includes: adding the raw material solution I into the raw material solution II at the flow rate of 400ml/min, heating for reaction, recovering a reaction solvent, adding water, uniformly mixing, filtering, washing with water, and collecting a filter cake.
Preferably, the step (a3) specifically includes: heating the raw material solution II to 70 ℃, adding the raw material solution I into the raw material solution II at the flow rate of 400ml/min, heating to 85 ℃, stirring, reacting for 18h, recovering a reaction solvent, adding water at the temperature of 40-50 ℃, stirring, filtering, washing with water until the pH value of an effluent liquid is 6-7, and collecting a filter cake.
Preferably, hydrochloric acid is added into the filtrate subjected to the fine filtration in the step (A5) to adjust the pH value to 5-6.
Preferably, the reaction solvent is a pyridine or amide solvent.
Preferably, the amide solvent is selected from any one of methylacetamide, ethylacetamide, dimethyloxamide, dimethylacetamide, or dimethylformamide.
Preferably, the amide solvent is N, N dimethylformamide.
Preferably, the reaction solvent is an amide solvent.
Preferably, the alkaline agent is an inorganic alkaline agent.
Preferably, the alkaline agent is selected from one or more of sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate and potassium carbonate solution.
Preferably, the alkaline agent is sodium hydroxide.
Preferably, the preparation method further comprises: extracting with ethanol after enzymolysis.
Preferably, the ethanol is 95% ethanol.
Preferably, the preparation method further comprises: filtering after enzymolysis treatment (I), refluxing and extracting by ethanol, filtering (II), collecting filtrate, crystallizing, standing, filtering (III), washing, collecting filter cake, drying and crushing.
Preferably, the filtration (I) is plate and frame filtration.
Preferably, the ethanol extraction process is ethanol reflux extraction.
Preferably, the ethanol extraction process is reflux extraction with 95% ethanol.
Preferably, the reflux extraction process has a reflux extraction time of 2 h.
Preferably, the crystallization process is specifically: adding cooling water, standing and crystallizing.
Preferably, the washing process is specifically: washing with water until no alcohol smell.
Preferably, the preparation method specifically comprises: filtering with plate-frame filter after enzymolysis, reflux extracting with ethanol, filtering (II), collecting filtrate, adding cooling water, standing for crystallization, filtering (III), washing with water until no alcohol smell exists, collecting filter cake, oven drying, and pulverizing.
The detailed description of the present application compared to the prior art is as follows
The application provides a method for preparing diosmin, which is used for preparing diosmin by enzymolysis, wherein the principle of diosmin enzymolysis is that rhamnose and glucose of diosmin are removed under the action of hesperidinase.
The preparation method of diosmetin does not generate a large amount of waste water and does not cause pollution to the environment.
The preparation method of diosmetin has the advantages of short time, simple post-treatment, low production cost and high yield, and the prepared diosmetin has high HPLC content, high purity and good product quality and is suitable for industrial production.
The invention screens the proper dosage of the hesperidinase, the optimal dosage is that the hesperidinase with 2 percent of the weight of the hesperidin (2.38 percent of the weight of the diosmin) is used for enzymolysis, the lower limit is 0.5 percent of the weight of the hesperidin (0.60 percent of the weight of the diosmin), and the upper limit is 20 percent of the weight of the hesperidin (23.81 percent of the weight of the diosmin). More than 20% by weight of hesperidin (23.81% by weight of diosmin) causes enzyme waste and increases production cost. When the amount of hesperidinase is small, the diosmin does not react completely.
The optimal enzymolysis temperature is 50 ℃, and the optimal enzymolysis time is 24 hours. The method has the advantages of easily controlled reaction conditions, simple operation and convenient treatment.
Detailed Description
In order to make those skilled in the art better understand the technical solution of the present invention, the following detailed description of the present invention is provided with reference to specific embodiments.
A method for preparing diosmetin comprises the following steps: diosmin is subjected to enzymolysis by using enzyme; the weight ratio of diosmin to enzyme is 100 (0.60-23.81).
Preferably, the preparation method specifically comprises: mixing diosmin, enzyme and water for enzymolysis; the weight ratio of the diosmin to the enzyme is 100 (0.60-23.81); the ratio of diosmin to water was 105 kg: 1m3。
Preferably, the enzyme is selected from one or more of hesperidinase, rhamnosidase, naringinase, glucosidase and pectinase. Preferably, the ratio of diosmin to enzyme by weight is 100: 2.38.
Preferably, the enzyme is hesperidinase.
Preferably, the enzymolysis temperature is 35-70 ℃, and the enzymolysis time is 12-72 h.
Preferably, the enzymolysis temperature is 40-50 ℃.
Preferably, the enzymatic hydrolysis temperature is 50 ℃.
Preferably, the enzymolysis time is 24 h.
Preferably, the preparation method further comprises: mixing the reaction solvent, hesperidin, alkaline reagent and iodine, and heating for reaction to obtain diosmin.
Preferably, the preparation method further comprises:
(A1) mixing iodine and a reaction solvent to obtain a raw material solution I;
(A2) mixing hesperidin with a reaction solvent to obtain a raw material solution II;
(A3) mixing the raw material solution I and the raw material solution II, heating for reaction, recovering a reaction solvent, adding water, uniformly mixing, filtering, washing with water, and collecting a filter cake;
(A4) mixing and dissolving the filter cake and an alkaline solution, and filtering by using a fine filter to obtain a fine-filtered filtrate;
(A5) and mixing the fine filtered filtrate with hydrochloric acid, crystallizing, filtering, and collecting a filter cake to obtain diosmin.
Preferably, the weight ratio of the hesperidin to the enzyme is 100 (0.5-20).
Preferably, the hesperidin to enzyme weight ratio is 100:2.
Preferably, the weight ratio of iodine to hesperidin is 1: 5 to 15.
Preferably, the weight ratio of iodine to hesperidin is 3: 10.
preferably, the ratio of the reaction solvent to the iodine used in the step (A1) is 1m3：75kg
Preferably, the ratio of the reaction solvent to the hesperidin used in the step (A2) is 1m3：200～300kg。
Preferably, the ratio of the reaction solvent to the hesperidin used in the step (A2) is 1m3：250kg。
Preferably, the step (a3) specifically includes: adding the raw material solution I into the raw material solution II at the flow rate of 400ml/min, heating for reaction, recovering a reaction solvent, adding water, uniformly mixing, filtering, washing with water, and collecting a filter cake.
Preferably, the step (a3) specifically includes: heating the raw material solution II to 70 ℃, adding the raw material solution I into the raw material solution II at the flow rate of 400ml/min, heating to 85 ℃, stirring, reacting for 18h, recovering a reaction solvent, adding water at the temperature of 40-50 ℃, stirring, filtering, washing with water until the pH value of an effluent liquid is 6-7, and collecting a filter cake.
Preferably, hydrochloric acid is added into the filtrate subjected to the fine filtration in the step (A5) to adjust the pH value to 5-6.
Preferably, the reaction solvent is a pyridine or amide solvent.
Preferably, the amide solvent is selected from any one of methylacetamide, ethylacetamide, dimethyloxamide, dimethylacetamide, or dimethylformamide.
Preferably, the amide solvent is N, N dimethylformamide.
Preferably, the reaction solvent is an amide solvent.
Preferably, the alkaline agent is an inorganic alkaline agent.
Preferably, the alkaline agent is selected from one or more of sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate and potassium carbonate solution.
Preferably, the alkaline agent is sodium hydroxide.
Preferably, the preparation method further comprises: extracting with ethanol after enzymolysis.
Preferably, the ethanol is 95% ethanol.
Preferably, the preparation method further comprises: filtering after enzymolysis treatment (I), refluxing and extracting by ethanol, filtering (II), collecting filtrate, crystallizing, standing, filtering (III), washing, collecting filter cake, drying and crushing.
Preferably, the filtration (I) is plate and frame filtration.
Preferably, the ethanol extraction process is ethanol reflux extraction.
Preferably, the ethanol extraction process is reflux extraction with 95% ethanol.
Preferably, the reflux extraction process has a reflux extraction time of 2 h.
Preferably, the crystallization process is specifically: adding cooling water, standing and crystallizing.
Preferably, the washing process is specifically: washing with water until no alcohol smell.
Preferably, the preparation method specifically comprises: filtering with plate-frame filter after enzymolysis, reflux extracting with ethanol, filtering (II), collecting filtrate, adding cooling water, standing for crystallization, filtering (III), washing with water until no alcohol smell exists, collecting filter cake, oven drying, and pulverizing.
The reagents used in the examples are commercially available reagents, 95% ethanol is 95% ethanol by volume.
Example 1
A method for preparing diosmetin comprises the following steps:
(A) preparation of diosmetin:
(A1)75kg iodine 1m3Dissolving a reaction solvent (N, N Dimethylformamide (DMF)) to obtain a raw material solution I for later use;
(A2) adding 250kg hesperidin into the mixture for 1m3A reaction solvent (N, N Dimethylformamide (DMF)) to obtain a raw material solution II;
(A3) heating the raw material solution II to 70 ℃, adding the raw material solution I into the raw material solution II at the flow rate of 400ml/min, heating to 85 ℃, stirring, reacting for 18 hours, recovering a reaction solvent, adding water at 40 ℃, stirring uniformly, filtering, washing with water until the pH value of an effluent liquid is 7, and collecting a filter cake;
(A4) and adding hydrochloric acid into the filtrate to adjust the pH value to 5, standing for crystallization, filtering, and collecting a filter cake to obtain 210kg of diosmin.
(B) Enzymolysis: adding 210kg of diosmin into the mixture for 2m3Stirring and mixing uniformly in water, adding hesperidin in an amount of 2% of the weight of hesperidin (2.38% of the weight of diosmin) for enzymolysis, and performing enzymolysis at 50 ℃ for 24 hours to obtain an enzymolysis product;
(C) refining: filtering zymolyte, collecting solid filter cake, and collecting solid with a volume of 2m3Dissolving with 95% ethanol, reflux extracting for 2h, filtering, collecting filtrate, recovering ethanol to obtain filtrate 200L, adding cooling water, crystallizing, standing for 8h, filtering, washing with water until no alcohol smell is produced, collecting filter cake, oven drying, and pulverizing to obtain refined diosmetin 95 kg.
The yield of diosmetin is 45 percent
HPLC external standard method for detecting diosmetin content
Chromatographic conditions are as follows: specification and model of the chromatographic column: c18: column temperature 40 ℃ C. 100X 4.6mm X3 um
Flow rate 1.5 ml/min; mobile phase: acetonitrile-glacial acetic acid-methanol-water; detection wavelength: 275nm
Diosmetin test samples: the refined diosmetin prepared in example 1.
Preparation of a test solution: weighing 25mg of diosmetin sample, adding dimethyl sulfoxide into a 25ml volumetric flask, carrying out ultrasonic treatment for about 5min to completely dissolve the sample, cooling to room temperature (15-35 ℃) and then metering to 25ml with dimethyl sulfoxide to obtain the diosmetin.
Diosmetin control: diosmetin work reference.
Preparation of a reference solution: weighing 25mg of diosmetin working reference substance, adding dimethyl sulfoxide into a 25ml volumetric flask, performing ultrasonic treatment for about 5min to completely dissolve a test sample, cooling to room temperature, and metering to 25ml with dimethyl sulfoxide to obtain the diosmetin.
The detection method comprises the following steps: and (3) balancing the chromatographic system for 30-60min under the chromatographic conditions, automatically sampling the solutions in sequence after the baseline is stable, and automatically integrating by an instrument and keeping the chromatogram.
And (3) calculating the content:
the content calculation formula is as follows:
Atest articleIs the peak area of the test sample;
Areference substancePeak area for control;
Mtest articleThe concentration is the concentration of the test solution, mg/ml;
Mreference substanceThe concentration of the reference solution is mg/ml;
Creference substance-is the nominal content of the control substance%
LOD is the loss on drying,%, of the test sample;
the refined diosmetin prepared in example 1 has an external standard HPLC content of 98% and a purity of 99%.
Example 2
This example differs from example 1 only in that: in this example, the reaction solvent was pyridine. (alternative is the reaction solvent example 1 (N, N Dimethylformamide (DMF))
Example 3
This example differs from example 1 only in that: the enzymolysis temperature is 55 ℃.
Example 4
This example differs from example 1 only in (B) enzymatic hydrolysis: adding 210kg of diosmin into the mixture for 2m3Stirring and mixing uniformly in water, adding hesperidin 0.3% (0.36% of diosmin) for enzymolysis, and performing enzymolysis at 50 deg.C for 24 hr to obtain zymolyte;
example 5
This example differs from example 1 only in (B) enzymatic hydrolysis: adding 210kg of diosmin into the mixture for 2m3Stirring in water, adding hesperidin 0.5 wt% (0.60 wt% of diosmin) of hesperidin, and performing enzyme treatmentHydrolyzing, and performing enzymolysis at 50 deg.C for 24 hr to obtain zymolyte.
Example 6
This example differs from example 1 only in (B) enzymatic hydrolysis: enzymolysis with rhamnosidase.
Example 7
This example differs from example 1 only in (B) enzymatic hydrolysis: enzymolysis with naringinase.
Example 8
This example differs from example 1 only in (B) enzymatic hydrolysis: enzymolysis with glucosidase.
Example 9
This example differs from example 1 only in (B) enzymatic hydrolysis: enzymolysis with pectinase.
Effect example 1
Diosmetin was prepared according to the preparation method of example 1, the yield of the reaction carried out under different dosages of hesperidinase, the content and purity detected by HPLC external standard method were measured, and the results are shown in Table 1
TABLE 1
The best usage amount of hesperidin is 2% (2.38% by weight of diosmin) for enzymolysis, the lower limit is 0.5% (0.60% by weight of diosmin) by weight of hesperidin, and the upper limit is 20% (23.81% by weight of diosmin) by weight of hesperidin. More than 20% by weight of hesperidin (23.81% by weight of diosmin) causes enzyme waste and increases production cost. When the amount of the diosmin is small, the reaction of the diosmin is incomplete, and the purity of the obtained product is low.
Effect example 2
Diosmetin was prepared according to the preparation method of example 1, the yield of the reaction carried out under different enzymolysis temperature and enzymolysis time conditions, the content and purity detected by HPLC external standard method were measured, and the results are shown in Table 2 (the amount of hesperidin used is 2% of the weight of hesperidin)
TABLE 2
The enzymolysis temperature and the enzymolysis time have obvious quality influence on the yield of the product and the content and the purity detected by an HPLC external standard method. The enzymolysis temperature is 35-70 ℃, and the enzymolysis time is 12-72 h. Preferably, the enzymolysis temperature is 40-50 ℃. The enzymolysis temperature is 50 ℃, the enzymolysis time exceeds 24 hours, and the content and purity detected by an HPLC external standard method are not obviously changed, so the preparation efficiency and the cost are considered, and the enzymolysis temperature is most preferably 50 ℃, and the enzymolysis time is 24 hours.
The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and these modifications and adaptations should be considered within the scope of the invention.
Claims (10)
1. A method for preparing diosmetin is characterized by comprising the following steps: diosmin is subjected to enzymolysis by using enzyme; the weight ratio of diosmin to enzyme is 100 (0.60-23.81).
2. The preparation method according to claim 1, wherein the preparation method specifically comprises: mixing diosmin, enzyme and water for enzymolysis; the weight ratio of the diosmin to the enzyme is 100 (0.6-23.81); the ratio of diosmin to water was 105 kg: 1m3。
3. The method according to claim 1, wherein the enzyme is one or more selected from the group consisting of hesperidinase, rhamnosidase, naringinase, glucosidase and pectinase.
4. The method of claim 1, wherein the ratio of diosmin to enzyme by weight is 100: 2.38.
5. The preparation method according to claim 1, wherein the enzymolysis temperature is 35-70 ℃ and the enzymolysis time is 12-72 h.
6. The preparation method according to claim 5, wherein the enzymolysis temperature is 50 ℃ and the enzymolysis time is 24 h.
7. The method of manufacturing according to claim 1, further comprising: mixing the reaction solvent, hesperidin, alkaline reagent and iodine, and heating for reaction to obtain diosmin.
8. The method of manufacturing according to claim 7, further comprising:
(A1) mixing iodine and a reaction solvent to obtain a raw material solution I;
(A2) mixing hesperidin with a reaction solvent to obtain a raw material solution II;
(A3) mixing the raw material solution I and the raw material solution II, heating for reaction, recovering a reaction solvent, adding water, uniformly mixing, filtering, washing with water, and collecting a filter cake;
(A4) mixing and dissolving the filter cake and an alkaline solution, and filtering by using a fine filter to obtain a fine-filtered filtrate;
(A5) and mixing the fine filtered filtrate with hydrochloric acid, crystallizing, filtering, and collecting a filter cake to obtain diosmin.
9. The method of manufacturing according to claim 1, further comprising: extracting with ethanol after enzymolysis.
10. The method of manufacturing according to claim 9, further comprising: filtering after enzymolysis treatment (I), extracting with ethanol, filtering (II), collecting filtrate, crystallizing, standing, filtering (III), washing, collecting filter cake, drying, and pulverizing.
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20080049174A (en) * | 2006-11-30 | 2008-06-04 | (주)아모레퍼시픽 | Preparing method of diosmetin from lemon peel and the cosmetic composition containing thereof as active ingredient |
| CN102070689A (en) * | 2011-01-25 | 2011-05-25 | 湖南圆通药业有限公司 | Method for producing diosmin |
| CN103819520A (en) * | 2014-02-14 | 2014-05-28 | 闻永举 | Method for preparing mono-glucoside through selective hydrolysis of flavone rutinoside or neohesperidoside |
| WO2014131995A1 (en) * | 2013-02-28 | 2014-09-04 | Les Laboratoires Servier | Method for the enzymatic synthesis of diosmetin derivatives |
| CN105732744A (en) * | 2016-04-29 | 2016-07-06 | 南京正大天晴制药有限公司 | Method for preparing green and economic diosmin |
| CN109180627A (en) * | 2018-09-06 | 2019-01-11 | 陕西嘉禾生物科技股份有限公司 | A kind of semisynthesis of diosmetin |
| CN109369591A (en) * | 2018-11-29 | 2019-02-22 | 陕西嘉禾生物科技股份有限公司 | A kind of synthetic method of diosmetin |
| CN111100104A (en) * | 2019-12-26 | 2020-05-05 | 陕西嘉禾药业有限公司 | Preparation method of diosmetin |
-
2021
- 2021-10-27 CN CN202111255615.5A patent/CN114317630A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20080049174A (en) * | 2006-11-30 | 2008-06-04 | (주)아모레퍼시픽 | Preparing method of diosmetin from lemon peel and the cosmetic composition containing thereof as active ingredient |
| CN102070689A (en) * | 2011-01-25 | 2011-05-25 | 湖南圆通药业有限公司 | Method for producing diosmin |
| WO2014131995A1 (en) * | 2013-02-28 | 2014-09-04 | Les Laboratoires Servier | Method for the enzymatic synthesis of diosmetin derivatives |
| CN103819520A (en) * | 2014-02-14 | 2014-05-28 | 闻永举 | Method for preparing mono-glucoside through selective hydrolysis of flavone rutinoside or neohesperidoside |
| CN105732744A (en) * | 2016-04-29 | 2016-07-06 | 南京正大天晴制药有限公司 | Method for preparing green and economic diosmin |
| CN109180627A (en) * | 2018-09-06 | 2019-01-11 | 陕西嘉禾生物科技股份有限公司 | A kind of semisynthesis of diosmetin |
| CN109369591A (en) * | 2018-11-29 | 2019-02-22 | 陕西嘉禾生物科技股份有限公司 | A kind of synthetic method of diosmetin |
| CN111100104A (en) * | 2019-12-26 | 2020-05-05 | 陕西嘉禾药业有限公司 | Preparation method of diosmetin |
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