CN103667401B - A kind of lipase-catalyzed online synthesis 6 " method of-O-lauroyl-Neohesperidin ester - Google Patents
A kind of lipase-catalyzed online synthesis 6 " method of-O-lauroyl-Neohesperidin ester Download PDFInfo
- Publication number
- CN103667401B CN103667401B CN201310392308.0A CN201310392308A CN103667401B CN 103667401 B CN103667401 B CN 103667401B CN 201310392308 A CN201310392308 A CN 201310392308A CN 103667401 B CN103667401 B CN 103667401B
- Authority
- CN
- China
- Prior art keywords
- reaction
- neohesperidin
- channel
- lauroyl
- ester
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000000034 method Methods 0.000 title claims abstract description 38
- 238000003786 synthesis reaction Methods 0.000 title claims abstract description 24
- 230000015572 biosynthetic process Effects 0.000 title claims abstract description 23
- 238000006243 chemical reaction Methods 0.000 claims abstract description 114
- 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 abstract description 48
- ARGKVCXINMKCAZ-UZRWAPQLSA-N neohesperidin 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)O3)O[C@H]3[C@@H]([C@H](O)[C@@H](O)[C@H](C)O3)O)=CC(O)=C2C(=O)C1 ARGKVCXINMKCAZ-UZRWAPQLSA-N 0.000 claims abstract description 44
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims abstract description 41
- MSXVEPNJUHWQHW-UHFFFAOYSA-N 2-methylbutan-2-ol Chemical compound CCC(C)(C)O MSXVEPNJUHWQHW-UHFFFAOYSA-N 0.000 claims abstract description 37
- 230000035484 reaction time Effects 0.000 claims abstract description 17
- GLVVKKSPKXTQRB-UHFFFAOYSA-N ethenyl dodecanoate Chemical compound CCCCCCCCCCCC(=O)OC=C GLVVKKSPKXTQRB-UHFFFAOYSA-N 0.000 claims abstract description 16
- 238000005917 acylation reaction Methods 0.000 claims abstract description 15
- 108010048733 Lipozyme Proteins 0.000 claims abstract description 14
- FCCDDURTIIUXBY-UHFFFAOYSA-N lipoamide Chemical compound NC(=O)CCCCC1CCSS1 FCCDDURTIIUXBY-UHFFFAOYSA-N 0.000 claims abstract description 14
- 239000007810 chemical reaction solvent Substances 0.000 claims abstract description 12
- 239000002994 raw material Substances 0.000 claims abstract description 12
- 239000012046 mixed solvent Substances 0.000 claims abstract description 10
- 239000000047 product Substances 0.000 claims description 10
- 239000000126 substance Substances 0.000 claims description 7
- 239000006227 byproduct Substances 0.000 claims description 3
- 230000001360 synchronised effect Effects 0.000 claims description 3
- 230000008901 benefit Effects 0.000 abstract description 5
- 230000009466 transformation Effects 0.000 description 24
- -1 flavonoid compound Chemical class 0.000 description 18
- 230000000052 comparative effect Effects 0.000 description 12
- 229930003935 flavonoid Natural products 0.000 description 11
- 235000017173 flavonoids Nutrition 0.000 description 11
- 230000002255 enzymatic effect Effects 0.000 description 10
- 230000032050 esterification Effects 0.000 description 9
- 238000005886 esterification reaction Methods 0.000 description 9
- 229940070765 laurate Drugs 0.000 description 9
- 239000005639 Lauric acid Substances 0.000 description 7
- 239000000376 reactant Substances 0.000 description 7
- 238000011160 research Methods 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical group CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 230000003078 antioxidant effect Effects 0.000 description 5
- 208000012839 conversion disease Diseases 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- 240000004307 Citrus medica Species 0.000 description 4
- 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 description 4
- 230000004071 biological effect Effects 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 4
- 238000010828 elution Methods 0.000 description 4
- 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 description 4
- 230000009467 reduction Effects 0.000 description 4
- 238000004809 thin layer chromatography Methods 0.000 description 4
- 241000193830 Bacillus <bacterium> Species 0.000 description 3
- 108091005658 Basic proteases Proteins 0.000 description 3
- 238000006555 catalytic reaction Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- POULHZVOKOAJMA-UHFFFAOYSA-M dodecanoate Chemical compound CCCCCCCCCCCC([O-])=O POULHZVOKOAJMA-UHFFFAOYSA-M 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- CQRYARSYNCAZFO-UHFFFAOYSA-N salicyl alcohol Chemical compound OCC1=CC=CC=C1O CQRYARSYNCAZFO-UHFFFAOYSA-N 0.000 description 3
- 240000006439 Aspergillus oryzae Species 0.000 description 2
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 2
- 206010007191 Capillary fragility Diseases 0.000 description 2
- 241000207199 Citrus Species 0.000 description 2
- 235000005979 Citrus limon Nutrition 0.000 description 2
- 244000248349 Citrus limon Species 0.000 description 2
- 235000001938 Citrus medica Nutrition 0.000 description 2
- 235000002555 Citrus medica var sarcodactylis Nutrition 0.000 description 2
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 108010084311 Novozyme 435 Proteins 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 230000001093 anti-cancer Effects 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 235000012000 cholesterol Nutrition 0.000 description 2
- 235000020971 citrus fruits Nutrition 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 235000014113 dietary fatty acids Nutrition 0.000 description 2
- 238000004821 distillation Methods 0.000 description 2
- POULHZVOKOAJMA-UHFFFAOYSA-N dodecanoic acid Chemical compound CCCCCCCCCCCC(O)=O POULHZVOKOAJMA-UHFFFAOYSA-N 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 229930195729 fatty acid Natural products 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 150000002430 hydrocarbons Chemical group 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- 230000004089 microcirculation Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 231100000614 poison Toxicity 0.000 description 2
- 230000007096 poisonous effect Effects 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- DKVBOUDTNWVDEP-NJCHZNEYSA-N teicoplanin aglycone Chemical compound N([C@H](C(N[C@@H](C1=CC(O)=CC(O)=C1C=1C(O)=CC=C2C=1)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)OC=1C=C3C=C(C=1O)OC1=CC=C(C=C1Cl)C[C@H](C(=O)N1)NC([C@H](N)C=4C=C(O5)C(O)=CC=4)=O)C(=O)[C@@H]2NC(=O)[C@@H]3NC(=O)[C@@H]1C1=CC5=CC(O)=C1 DKVBOUDTNWVDEP-NJCHZNEYSA-N 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 241000040710 Chela Species 0.000 description 1
- 102000007330 LDL Lipoproteins Human genes 0.000 description 1
- 108010007622 LDL Lipoproteins Proteins 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 241000235403 Rhizomucor miehei Species 0.000 description 1
- 241001093501 Rutaceae Species 0.000 description 1
- ZONYXWQDUYMKFB-UHFFFAOYSA-N SJ000286395 Natural products O1C2=CC=CC=C2C(=O)CC1C1=CC=CC=C1 ZONYXWQDUYMKFB-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 230000000118 anti-neoplastic effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000003035 anti-peroxidant effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 235000021466 carotenoid Nutrition 0.000 description 1
- 150000001747 carotenoids Chemical class 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 230000000536 complexating effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 150000005690 diesters Chemical class 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 1
- 230000009881 electrostatic interaction Effects 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 230000009144 enzymatic modification Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 229930003949 flavanone Natural products 0.000 description 1
- 150000002207 flavanone derivatives Chemical class 0.000 description 1
- 235000011981 flavanones Nutrition 0.000 description 1
- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 229930182486 flavonoid glycoside Natural products 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
- 229940094952 green tea extract Drugs 0.000 description 1
- 235000020688 green tea extract Nutrition 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000005311 nuclear magnetism Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 238000012797 qualification Methods 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a kind of lipase-catalyzed online synthesis 6 " method of-O-lauroyl-Neohesperidin ester; described method is: take mol ratio as Neohesperidin and the vinyl laurate of 1:1 ~ 12 be raw material; with 0.5 ~ 1.0g Lipozyme? does is RMIM catalyzer; with the mixed solvent of tertiary amyl alcohol and DMSO for reaction solvent; by Lipozyme? RMIM is uniformly filled in the reaction channel of microfluidic channel reactor, the reaction channel internal diameter of described microfluidic channel reactor is 0.8 ~ 2.4mm, and reaction channel length is 0.5 ~ 1.0m; Raw material and reaction solvent are passed in reaction channel continuously and carries out acylation reaction, controlling acylation reaction temperature is 40 ~ 55 DEG C, the acylation reaction time is 20 ~ 40min, collects reaction solution online, and reaction solution obtains 6 through conventional aftertreatment "-O-lauroyl-Neohesperidin ester.The present invention has the advantage that the reaction times is short, selectivity is high and productive rate is high.
Description
(1) technical field
The present invention relates to a kind of lipase-catalyzed online controlled selectivity synthesis 6 " method of-O-lauroyl-Neohesperidin ester.
(2) background technology
Neohesperidin (Neohesperidin) be also neohesperidin, and be a kind of flavanone kind composition, be extensively present in the plants such as the dried immature fruit of citron orange, citrus, lemon, Buddha's hand, especially in the pericarp of rutaceae, content is higher.There is good anticancer, antibacterial, anti-oxidant, decreasing cholesterol, reduction capillary fragility, improve the multiple biological activitys such as microcirculation, be usually used in the fields such as medicine, makeup, food.
The various biological activitys of flavonoid compound are all based on anti-oxidant activity according to the study, relevant with Green Tea Extract or anti peroxidation of lipid.Special structure gives the chemical property of its a series of uniqueness, if with many kinds of metal ions generation complexing or electrostatic interaction, there is reduction and catch the characteristic of free radical and many derivative reactions active etc.Flavonoid compound suppresses low-density lipoprotein LDL oxidation and suppresses the research of carotenoid and unsaturated fatty acids acid oxidase to show, aglycon has more outstanding anti-oxidant activity than glucosides.This is because aglycon lipotropy is strong, can embed in the kernel of microbial film hydrophobic layer and play a role.The poly-hydroxy of flavonoid glycoside compound and glycosyl structure make it have certain wetting ability, and this will certainly affect anti-oxidant activity.Flavonoid compound is modified by esterification, and introducing long hydrocarbon chain in the molecule can increase the fat-soluble of it, thus increases with the consistency of fat and improve oxidation-resistance.So, Neohesperidin is as applying one of maximum flavonoid compound, and the Neohesperidin fatty acid ester researching and developing its physiologically active stronger will have a good application prospect.
Research abroad for neohesperidin mainly concentrates on the aspects such as extracting and developing, qualification and clinical antineoplastic.The domestic research for neohesperidin mainly concentrates on In Vitro Bacteriostasis, antioxidation in vitro, falls clinical pharmacology and the health-care effect aspects such as blood ester.Few for the research of the modification application this respect of Neohesperidin both at home and abroad, mainly make its modification with the method process of chemistry, make exactly hesperidine and some metal ion chela with, to strengthen its biological activity; Carry out modification by enzymatic reaction method in addition in addition, object improves its fat-soluble and thermotolerance.Enzymatic modification because reaction conditions is gentle, selectivity comparatively advantages of higher be more and more concerned.At present, Neohesperidin is modified by the esterification of enzymatic regioselectivity, and introducing long hydrocarbon chain in the molecule can increase the fat-soluble of it, thus increases with the consistency of fat and improve oxidation-resistance.The mild condition although enzymatic reaction responds, the advantage that selectivity is good, but enzymic catalytic reaction is having some limitations property also, restriction between the activity of such as enzyme and the polarity of organic solvent, restriction between the polarity of organic solvent and the solvability of reaction substrate, reaction times longer, transformation efficiency not high-technology bottleneck.Therefore, Neohesperidin enzymatic selective esterification new technology is found significant for the production of development Neohesperidin fatty acid ester.
Micro-fluidic (Microfluidics) manipulates to receive to rise to technology and the science that skin rises volume fluid in micron scale construction, is the new cross discipline emerged rapidly nearly ten years.Current, the development of micro-fluidic has surmounted the original object being mainly analytical chemistry service greatly, and is becoming the important technological platform of whole chemistry subject, life science, instrumental science and even information science new round innovation research.
After Harrison seminar in 1997 has delivered the document of first section synthetic compound in micro-fluidic chip microreactor, micro-fluidic chip reactor has been successfully used to multiple organic synthesis, and illustrates application prospect widely.Along with the development of microring array, micro-reacting tcchnology in micro-fluidic chip, carry out building-up reactions in the chips and become one of the study hotspot in micro-fluidic chip field.
Compare with conventional chemical reactor, micro passage reaction not only has makes the diffusion length between reactant greatly shorten, and mass transfer velocity is fast; The easy control of reaction conditions such as reactant ratio, temperature, reaction times and flow velocity, side reaction is less; Need reactant consumption very micro-, not only can reduce the consumption of costliness, poisonous, adverse reaction thing, the environmental pollutant produced in reaction process are also few, are the technology of a kind of environmental friendliness, study on the synthesis novel substance.
At present, more Chinese scholars is had to be studied the Enzyme catalyzed synthesis of Neohesperidin ester in organic medium, but the method often needs the longer reaction times (24h), and reaction transformation efficiency and selectivity not high, utilizing microflow control technique to explore new building-up reactions, is study hotspot international and domestic now in order to solve the technical bottleneck of existing synthetic method.Our research group has also carried out systematic study to the enzymatic selectivity synthesis of saccharide compound in micro-fluidic reactor early stage, finds that the enzymatic selectivity synthesis utilizing microflow control technique to carry out saccharide compound has certain feasibility.And Neohesperidin is the sugary flavonoid compound of a class, derive from the plants such as the dried immature fruit of citron orange, citrus, lemon, Buddha's hand, there is good anticancer, antibacterial, anti-oxidant, decreasing cholesterol, reduction capillary fragility, improve the multiple biological activitys such as microcirculation.The regioselectivity esterification of Neohesperidin is modified and not only can improve bioavailability, its application in fields such as medicine, food can also be expanded.And Neohesperidin is the sugary flavonoid compound of a class, we can utilize the enzymatic regioselectivity esterification of Research foundation to Neohesperidin in early stage to study.Therefore " method of-O-lauroyl-Neohesperidin ester is intended to the online controlled method for selective synthesis of the Neohesperidin lauric acid monoester finding a kind of high-efficiency environment friendly to we have studied lipase-catalyzed selectivity synthesis 6 in microfluidic channel reactor.
(3) summary of the invention
The technical problem to be solved in the present invention is to provide lipase-catalyzed selectivity synthesis 6 in a kind of microfluidic channel reactor, and " novel process of-O-lauroyl-Neohesperidin ester, has the advantage that the reaction times is short, selectivity is high and productive rate is high.
For solving the problems of the technologies described above, the present invention adopts following technical scheme:
A kind of lipase-catalyzed online synthesis 6 " method of-O-lauroyl-Neohesperidin ester, described method adopts microfluidic channel reactor, described microfluidic channel reactor comprises syringe pump, syringe, reaction channel and product collector, described syringe is installed in syringe pump, be communicated with the entrance of reaction channel by first interface, described product collector passes through the outlet of the second interface and reaction channel, described reaction channel internal diameter is 0.8 ~ 2.4mm, and reaction channel length is 0.5 ~ 1.0m, described method comprises: the Neohesperidin being 1:1 ~ 1:12 with the ratio of amount of substance and vinyl laurate are for raw material, with 0.5 ~ 1.0g Lipozyme RMIM for catalyzer, with the mixed solvent of tertiary amyl alcohol and methyl-sulphoxide (DMSO) for reaction solvent, Lipozyme RMIM is uniformly filled in reaction channel, raw material and reaction solvent are placed in syringe, raw material and reaction solvent pass into continuously in reaction channel and carry out acylation reaction by syringe under the promotion of syringe pump, controlling acylation reaction temperature is 40 ~ 55 DEG C, the acylation reaction time is 20 ~ 40min, reaction solution is collected online by product collector, reaction solution is through the obtained 6 "-O-lauroyl-Neohesperidin esters of aftertreatment.
In the microfluidic channel reactor that the present invention adopts, described syringe number can be one or more, depending on concrete reaction requirement.Such as, when use two syringes, T-shaped or Y type interface can be adopted to be connected with the entrance of reaction channel, different reactants is introduced from two entrances, conflux and enter public reaction channel, increased with probability of collision by reactant molecule contact in microchannel, make two strands of reaction solution streams mix in public reaction channel and react.Multiple syringe synchronously promotes by syringe pump.
Described microfluidic channel reactor also comprises thermostat container, and described reaction channel is placed in thermostat container, and such as reaction channel is enclosed in case by thermostat container, effectively can control temperature of reaction with this.Described thermostat container can require to select voluntarily according to temperature of reaction, such as constant temperature water box etc.
The present invention does not limit for the material of reaction channel, and recommendation is green, the material of environmental protection, such as silicone tube; Shape for reaction channel is preferably curved shape, can ensure at the uniform velocity stable the passing through of reaction solution.
The present invention, in implementation process, first can dissolve Neohesperidin with the mixed solvent (tertiary amyl alcohol: DMSO=4:1) of tertiary amyl alcohol and DMSO, as long as its consumption ensures that Neohesperidin can fully dissolve, and is loaded in syringe for subsequent use; Only dissolve vinyl laurate with nontoxic tertiary amyl alcohol, be loaded in another syringe for subsequent use; Then under the synchronous promotion of syringe pump (such as PHD2000 syringe pump), make raw material and reaction solvent pass in reaction channel react.Therefore, in the present invention, in reaction solvent, tertiary amyl alcohol and DMSO volume ratio are greater than 4:1, react compared to the shaking table of routine, can be down to minimum by the usage quantity of poisonous DMSO.
Comparatively preferred, method of the present invention comprises the following steps:
First dissolve Neohesperidin with the mixed solvent that volume ratio is the tertiary amyl alcohol of 4:1 and DMSO, be loaded in syringe for subsequent use; Dissolve vinyl laurate with tertiary amyl alcohol, be loaded in another syringe for subsequent use; Two syringes are connected with the entrance of reaction channel by Y type interface, then raw material and reaction solvent are passed in reaction channel under the synchronous promotion of syringe pump and carry out acylation reaction.
When use two syringes respectively to reaction channel inject be dissolved with the tertiary amyl alcohol of Neohesperidin and the mixing solutions of DMSO and be dissolved with the tert-amyl alcohol solution of vinyl laurate time, volumetric usage for dissolving the tertiary amyl alcohol of vinyl laurate is preferably equal with the volumetric usage of dissolving the tertiary amyl alcohol of Neohesperidin and the mixed solvent of DMSO, so that the consistence of two kinds of material molar ratios when ensureing to enter reaction channel.
The volumetric usage of the mixed solvent of described tertiary amyl alcohol and DMSO for dissolving Neohesperidin counts 10 ~ 25mL/mmol with the amount of substance of Neohesperidin usually.
In the present invention, the commodity that described Lipozyme RMIM uses letter (novozymes) company of Novi to produce, being a kind of lipase obtained from Rhizomucormiehei, is produce through submerged fermentation with a kind of gene-modified aspergillus oryzae (Aspergillusoryzae) microorganism.LipozymeRMIM is granular product, and granularity is 0.2-0.6mm.
Lipozyme RMIM is uniformly filled in reaction channel by the inventive method, is directly evenly fixed in reaction channel by granular catalyzer by mechanical means.
Further, the mol ratio of described Neohesperidin and vinyl laurate is preferably 1:8 ~ 10, most preferably is 1:8.
Further, described acylation reaction temperature is preferably 52 ~ 55 DEG C, most preferably is 52 DEG C.
Further, the described acylation reaction time is preferably 30 ~ 40min, most preferably is 35min.Regulate the flow velocity of reaction channel inner fluid by regulating syringe pump and then regulate the residence time of raw material in reaction channel, i.e. the reaction times.
Reaction product of the present invention can be collected online, gained reaction solution can obtain 6 by conventional post-treating method "-O-lauroyl-Neohesperidin ester.Described conventional post-treating method can be: the underpressure distillation of gained reaction solution is except desolventizing, residuum is separated through silica gel column chromatography, with 200-300 order silica gel wet method dress post, elution reagent is ethyl acetate, methyl alcohol, water volume ratio are the mixed solvent of 25:2.5:0.6, wet method upper prop after residuum dissolves with a small amount of elution reagent, collect elutriant, TLC follows the tracks of wash-out process simultaneously, the elutriant containing single product obtained is merged evaporate to dryness, flaxen monoesters crystal can be obtained, be 6 "-O-lauroyl-Neohesperidin ester.
The reaction channel internal diameter that the present invention uses is 2mm, and reaction channel is long is 1.0m.Reaction channel internal diameter and length can affect rate of flow of fluid in reaction channel and the residence time, but do not cause direct impact to reaction itself.
In the present invention, although there is the hydroxyl of 6 similar nature in Neohesperidin, but "-O-lauroyl-Neohesperidin ester has higher transformation efficiency and selectivity to lipase-catalyzed selectivity synthesis 6 in micro-fluidic micro passage reaction; the transformation efficiency of diester is very low; almost do not have, the content 100% of the monoesters obtained by column chromatography for separation.The structure warp of product
1hNMR confirms.This shows that the enzymatic selectivity synthesis of Neohesperidin laurate in micro-fluidic micro passage reaction has good reaction conversion ratio and selectivity, can realize high monoesters rate.
Compared with prior art, beneficial effect of the present invention is: selectivity synthesis Neohesperidin-6 in microfluidic channel reactor of the present invention "-O-laurate, this method not only shortens the reaction times widely, and has high transformation efficiency and reaction preference; Reduce the usage quantity of DMSO simultaneously, there is environment-friendly advantage.
(4) accompanying drawing explanation
Fig. 1 is the structural representation of the microfluidic channel reactor that the embodiment of the present invention adopts.
(5) embodiment
With specific embodiment, protection scope of the present invention is described further below, but protection scope of the present invention is not limited thereto:
The structural reference Fig. 1 of the microfluidic channel reactor that the embodiment of the present invention uses, comprises a syringe pump (not shown), two syringes 1, reaction channel 3, constant temperature water box (5, only show its floor map) and product collector 4; Two syringes 1 are installed in syringe pump, be communicated with the entrance of reaction channel 3 by a Y type interface, described reaction channel 3 is placed in constant temperature water box 5, temperature of reaction is controlled by constant temperature water box 5, the internal diameter 2.0mm of described reaction channel 3, pipe range 1m, the outlet of described reaction channel 3 is communicated with product collector 4 by the second interface.
Embodiment 1:6 " the synthesis of-O-lauroyl-Neohesperidin ester
Neohesperidin (0.49mmol) is dissolved in 10mL tertiary amyl alcohol with reference to figure 1. by equipment therefor: in the mixed solvent of DMSO=4:1 (v/v), vinyl laurate (3.92mmol) is dissolved in 10mL tertiary amyl alcohol, is then loaded in 10mL syringe for subsequent use respectively.0.87g Lipozyme RMIM is uniformly filled in the reaction channel of microfluidic channel reactor, and under PHD2000 syringe pump promotes, two-way reaction solution is respectively with totally 8.91 μ Lmin
-1flow velocity entered in reaction channel by " Y " joint and react, temperature of reactor is controlled at 52 DEG C by constant temperature water box, syringe pump showing reaction solution actual volume in reaction channel is 314 microlitres, reaction solution is continuous flow reaction 35min in reaction channel, and reaction result is by thin-layer chromatography TLC tracing detection.
Reaction solution is collected online by product collector, underpressure distillation is except desolventizing, with 200-300 order silica gel wet method dress post, elution reagent is ethyl acetate: methyl alcohol: water volume ratio=25:2.5:0.6, post height 35cm, column diameter 4.5cm, wet method upper prop after sample dissolves with a small amount of elution reagent, flow velocity 2mLmin collected by elutriant
-1, simultaneously TLC follows the tracks of wash-out process, the elutriant containing single product obtained is merged evaporate to dryness, obtains flaxen monoesters crystal, obtain Neohesperidin-6 "-laurate is 6 "-O-lauroyl-Neohesperidin ester.HPLC detects Neohesperidin transformation efficiency 92%, the total content of the Neohesperidin ester of the content/generation of Neohesperidin-6 selectivity of "-the laurate (Neohesperidin-6 "-laurate) be 100%.
Nuclear-magnetism characterization result is as follows:
1h-NMR (DMSO-d6, δ, ppm): Neohesperidin-6 "-laurate: 12.03 (s, H
5), 9.12 (s ,-OH
3 '), 6.95 (s ,-OH
5 '), 6.92 (d, 1H, J=2.9Hz, H
6 '), 6.87 (m, 1H, H
2 '), 6.10 (d, 1H, J=2.2Hz, H
8), 6.06 (d, 1H, J=2.2Hz, H
6), 5.49 (m, 1H, H
2), 5.44 (m, 1H, H
1 "), 5.41 (m, 1H, 2 " '-OHofneohesperidin), 5.16 (m, 1H, H
1 " '), 5.10 (s, 1H, 4 " '-OHofneohesperidin); 4.74 (d, 1H, J=4.8Hz, 4 "-OHofneohesperidin), 4.69 (d, 1H, J=4.4Hz, 3 "-OHofneohesperidin), 4.50 (d, 1H; J=5.8Hz, 3 " '-OHofneohesperidin), 4.26 (d, 1H, J=11.9Hz, H
6 "acylated), 4.03 (m, 1H, H
6 "acylated), 3.77 (s, 3Hof-OCH
3ofphenyl), 3.73-3.14 (8H, Hofrhamnoglucosyl), 3.22 (m, 1H, H
3), 2.73 (d, 1H, J=3.2Hz, H
3), 2.22 (m, 2H ,-CH
2-CO-), 1.17 (m, 3HofCH
3ofrhamnosyl), 1.26 (m, 18Hof (CH
2)
9oflauroyl), 0.85 (t, 3H, J=7.0Hz, CH
3).
IR(KBr,cm
-1):3415(OH),1738(C=O),1647(C=C).
ESI-MS(m/z):828(M
1+2H
2O-H)
-,M
1correspondingexactlytothemolecularweightof6″-O-lauroyl–neohesperidin.
Embodiment 2-5
Change the temperature of microfluidic channel reactor, other are with embodiment 1, and reaction result is as shown in table 1:
Table 1: temperature is on the impact of reaction
The result of table 1 shows, when flow velocity is 8.91 μ Lmin
-1reaction times is when being 35min, react the rising with temperature, transformation efficiency also obviously raises, and when temperature of reaction reaches 52 DEG C, the transformation efficiency of reaction and selectivity are all best, if now continue to heat up, the reduction of enzymic activity will be caused, thus result in the transformation efficiency of reaction and selectivity decreases, thus in the present invention in micro-fluidic micro passage reaction the optimal reaction temperature of Neohesperidin laurate be 52 DEG C.
Embodiment 6-10
The substrate mol ratio changing vinyl laurate and Neohesperidin in micro-fluidic micro passage reaction is 1:1(embodiment 6), 4:1(embodiment 7), 6:1(embodiment 8), 10:1(embodiment 9), 12:1(embodiment 10), the consumption of Neohesperidin is that 0.49mmol is constant, changes the consumption of vinyl laurate.Other are with embodiment 1, and result is as shown in table 2.
Table 2: the impact that Neohesperidin and the comparison of vinyl laurate substrate are reacted
The result of table 2 shows, along with the increase of reactant vinyl laurate, the transformation efficiency of reaction also along with increase, when substrate than for 8:1 time, the transformation efficiency of reaction and selectivity optimum, Neohesperidin has substantially quantitatively transformed completely in order to Neohesperidin-6 "-laurate.If now continue the consumption increasing reactant vinyl laurate, the transformation efficiency of reaction and selectivity will be caused to reduce, and thus, the best substrate of this reaction is than being 8:1, under the reaction conditions, Neohesperidin transforms substantially completely in order to Neohesperidin-6 "-O-laurate.
Embodiment 11-14
Changing the reaction times in micro-fluidic micro passage reaction is 20min(embodiment 11), 25min(embodiment 12), 30min(embodiment 13), 40min(embodiment 14), other are with embodiment 1, and result is as shown in table 3.
Table 3: the reaction times is on reaction conversion ratio and optionally affect
| Embodiment | Reaction times [min] | Transformation efficiency [%] | Selectivity [%] |
| 11 | 20 | 20 | 100 |
| 12 | 25 | 60 | 100 |
| 13 | 30 | 85 | 100 |
| 1 | 35 | 92 | 100 |
| 14 | 40 | 86 | 99 |
The result of table 3 shows, the Neohesperidin-6 that 20min can obtain 20% is carried out in reaction, and "-lauric acid monoester, selectivity is now 100%, and Neohesperidin is be converted into Neohesperidin-6 "-lauric acid monoester completely substantially.Along with the increase in reaction times, the transformation efficiency of reaction increases gradually, and when 35min is carried out in reaction, the transformation efficiency of reaction and selectivity optimum, "-lauric acid monoester transformation efficiency can reach 92% to Neohesperidin-6, and reaction preference also can reach 100%.If now continue to extend the reaction times, can cause reaction conversion ratio on the contrary and optionally reduce, thus, " Best Times of-lauric acid monoester synthesis is 35min to Neohesperidin-6 in microfluidic channel reactor.
Comparative example 1 ~ 3
Change the catalyzer in micro-fluidic micro passage reaction, change Lipozyme TLIM(comparative example 1 respectively into), lipase Novozym435(comparative example 2), bacillus alkaline protease (comparative example 3), other are with embodiment 1, and result is as shown in table 4.
Table 4: different enzyme is on reaction conversion ratio and optionally affect
| Enzyme source | Transformation efficiency [%] | Selectivity [%] | |
| Comparative example 1 | Lipozyme TLIM | 50 | 100 |
| Comparative example 2 | Novozym435 | 30 | 100 |
| Comparative example 3 | Bacillus alkaline protease | 8 | 100 |
| Embodiment 1 | Lipozyme RMIM | 92 | 100 |
The result of table 4 shows, for the regioselectivity esterification of enzymatic Neohesperidin in microfluidic channel reactor, different enzymes has fairly obvious impact to reaction." transformation efficiency of-lauric acid monoester is 50% to utilize Lipozyme TLIM catalyzed reaction, Neohesperidin-6.And utilizing this reaction of bacillus alkaline protease catalysis, " transformation efficiency of-lauric acid monoester is only 8% to Neohesperidin-6.From the result of table 4, for the regioselectivity esterification of enzymatic Neohesperidin in microfluidic channel reactor, the most effective catalyzer is Lipozyme RMIM, and the transformation efficiency of Neohesperidin is 92%, and selectivity is 100%.
Comparative example 4 ~ 5
Change different types of flavonoid compound in micro-fluidic micro passage reaction, the Neohesperidin (0.49mmol) of embodiment 1 is changed into respectively hesperidine (comparative example 4), saligenin (comparative example 5), other with embodiment 1, result is as shown in table 5.
Table 5: different flavonoid compound is on reaction conversion ratio and optionally affect
| Flavone derivative | Transformation efficiency [%] | Selectivity [%] | |
| Comparative example 4 | Hesperidine | 2 | 100 |
| Comparative example 5 | Saligenin | 10 | 100 |
| Embodiment 1 | Neohesperidin | 92 | 100 |
The result of table 5 shows, for the regioselectivity esterification of enzymatic flavonoid compound in microfluidic channel reactor, different flavonoid compounds has different reaction results.Under the same reaction conditions, transformation efficiency is only 2% to hesperidine, and reaction and difficulty thereof, react hardly.The reaction result of saligenin is also undesirable, and transformation efficiency is only 10%.From the result of table 5, for the regioselectivity esterification of enzymatic flavonoid compound in microfluidic channel reactor, Neohesperidin can obtain more satisfactory reaction result, and the transformation efficiency of reaction can reach 92%, and selectivity is 100%.
Claims (6)
1. a lipase-catalyzed online synthesis 6 " method of-O-lauroyl-Neohesperidin ester, it is characterized in that described method adopts microfluidic channel reactor, described microfluidic channel reactor comprises syringe pump, syringe, reaction channel and product collector, described syringe is installed in syringe pump, be communicated with the entrance of reaction channel by first interface, described product collector passes through the outlet of the second interface and reaction channel, described reaction channel internal diameter is 0.8 ~ 2.4mm, and reaction channel length is 0.5 ~ 1.0m, described method comprises: the Neohesperidin being 1:8 ~ 10 with the ratio of amount of substance and vinyl laurate are for raw material, with 0.5 ~ 1.0g Lipozyme RMIM for catalyzer, with the mixed solvent of tertiary amyl alcohol and methyl-sulphoxide for reaction solvent, Lipozyme RMIM is uniformly filled in reaction channel, raw material and reaction solvent are placed in syringe, raw material and reaction solvent pass into continuously in reaction channel and carry out acylation reaction by syringe under the promotion of syringe pump, controlling acylation reaction temperature is 52 ~ 55 DEG C, the acylation reaction time is 30 ~ 40min, reaction solution is collected online by product collector, reaction solution is through the obtained 6 "-O-lauroyl-Neohesperidin esters of aftertreatment.
2. lipase-catalyzed online synthesis 6 as claimed in claim 1 " method of-O-lauroyl-Neohesperidin ester; it is characterized in that: described method comprises the following steps: first to dissolve Neohesperidin with the mixed solvent that volume ratio is the tertiary amyl alcohol of 4:1 and DMSO, is loaded in syringe for subsequent use; Dissolve vinyl laurate with tertiary amyl alcohol, be loaded in another syringe for subsequent use; Two syringes are connected with the entrance of reaction channel by Y type interface, then raw material and reaction solvent are passed in reaction channel under the synchronous promotion of syringe pump and carry out acylation reaction.
3. " method of-O-lauroyl-Neohesperidin ester, is characterized in that: described microfluidic channel reactor comprises thermostat container, and described reaction channel is placed in thermostat container in lipase-catalyzed online synthesis 6 as claimed in claim 1.
4. " method of-O-lauroyl-Neohesperidin ester, is characterized in that: described microfluidic channel reactor comprises thermostat container, and described reaction channel is placed in thermostat container in lipase-catalyzed online synthesis 6 as claimed in claim 2.
5. " method of-O-lauroyl-Neohesperidin ester, is characterized in that: described Neohesperidin is 1:8 with the ratio of the amount of substance of vinyl laurate in lipase-catalyzed online synthesis 6 as claimed in claim 1.
6. " method of-O-lauroyl-Neohesperidin ester, is characterized in that: described acylation reaction temperature is 52 DEG C, and the described acylation reaction time is 35min in lipase-catalyzed online synthesis 6 as claimed in claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310392308.0A CN103667401B (en) | 2013-09-02 | 2013-09-02 | A kind of lipase-catalyzed online synthesis 6 " method of-O-lauroyl-Neohesperidin ester |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310392308.0A CN103667401B (en) | 2013-09-02 | 2013-09-02 | A kind of lipase-catalyzed online synthesis 6 " method of-O-lauroyl-Neohesperidin ester |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103667401A CN103667401A (en) | 2014-03-26 |
| CN103667401B true CN103667401B (en) | 2015-11-25 |
Family
ID=50306169
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310392308.0A Active CN103667401B (en) | 2013-09-02 | 2013-09-02 | A kind of lipase-catalyzed online synthesis 6 " method of-O-lauroyl-Neohesperidin ester |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103667401B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107955822A (en) * | 2017-12-21 | 2018-04-24 | 浙江工业大学 | A kind of lipase-catalyzed online synthesis S-（4- methyl-benzyls）The method of laurate thioesters |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103184253A (en) * | 2011-12-31 | 2013-07-03 | 浙江工业大学 | Method of using lipase to catalyze and synthesize mannose-6-laurate on line |
| CN103184252A (en) * | 2011-12-31 | 2013-07-03 | 浙江工业大学 | Method of using lipase to catalyze and synthesize glucose-6-laurate on line |
| CN103184256A (en) * | 2011-12-31 | 2013-07-03 | 浙江工业大学 | Method for on-line synthesizing saccharose-6-laurate by lipase catalysis |
-
2013
- 2013-09-02 CN CN201310392308.0A patent/CN103667401B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103184253A (en) * | 2011-12-31 | 2013-07-03 | 浙江工业大学 | Method of using lipase to catalyze and synthesize mannose-6-laurate on line |
| CN103184252A (en) * | 2011-12-31 | 2013-07-03 | 浙江工业大学 | Method of using lipase to catalyze and synthesize glucose-6-laurate on line |
| CN103184256A (en) * | 2011-12-31 | 2013-07-03 | 浙江工业大学 | Method for on-line synthesizing saccharose-6-laurate by lipase catalysis |
Non-Patent Citations (2)
| Title |
|---|
| Biocatalytic preparation of acylated derivatives of flavonoid glycosides enhances their antioxidant and antimicrobial activity;F. Mellou等;《Journal of Biotechnology》;20051231(第116期);295-304 * |
| New progress in biocatalysis and biotransformation of flavonoids;Anming Wang等;《Journal of Medicinal Plants Research》;20100518;第4卷(第10期);847-856 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103667401A (en) | 2014-03-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103667402B (en) | A kind of lipase-catalyzed online synthesis 6 " method of-O-lauroyl-naringin ester | |
| CN103667396B (en) | A kind of lipase-catalyzed online synthesis 6 " method of-O-lauroyl-naringin dihydrochalcone ester | |
| CN103184249B (en) | Method for on-line synthesizing glucose-6-palmitate by lipase catalysis | |
| CN103184257B (en) | Method for on-line synthesizing sucrose-6-acetate catalyzed by lipase | |
| CN103184252B (en) | Method of using lipase to catalyze and synthesize glucose-6-laurate on line | |
| CN103184251B (en) | Method for on-line synthesizing glucose-6-acetate catalyzed by lipase | |
| CN103184253B (en) | Method of using lipase to catalyze and synthesize mannose-6-laurate on line | |
| CN103184255B (en) | Method of using lipase to catalyze and synthesize galactose-6-acetate on line | |
| CN105838600B (en) | A kind of method of 5 ' O palmityl uridines of lipase-catalyzed online synthesis | |
| CN107475330A (en) | A kind of method of lipase-catalyzed online synthesis N (5 glucose ester valeryl) metoprolol | |
| CN103184256A (en) | Method for on-line synthesizing saccharose-6-laurate by lipase catalysis | |
| CN103182277B (en) | Microfluidic channel reactor and the application in synthesis of sucrose-6-palmitate thereof | |
| CN107384992A (en) | A kind of method of lipase-catalyzed online synthesis 5 '-O- lauroyl -5-methyl-uridin | |
| CN107384991A (en) | A kind of method of 5 '-O- ethene adipyl uridines of lipase-catalyzed online synthesis | |
| CN103667400B (en) | A kind of lipase-catalyzed online synthesis 6 " method of-O-palmityl-naringin ester | |
| CN103667393B (en) | A kind of lipase-catalyzed online synthesis 6 " method of-O-palmityl-neohesperidin dihydrochalcone ester | |
| CN107384781A (en) | A kind of method of lipase-catalyzed online synthesis 5 '-O- ethene adipyls -5-methyl-uridin | |
| CN107384782A (en) | A kind of method of lipase-catalyzed online synthesis 5 '-O- ethene adipyls -5-FUD | |
| CN107488690A (en) | A kind of method of lipase-catalyzed online synthesis N (5 glucose ester valeryl) mexiletine | |
| CN103667394B (en) | A kind of lipase-catalyzed online synthesis 6 " method of-O-lauroyl-neohesperidin dihydrochalcone ester | |
| CN103667399B (en) | A kind of lipase-catalyzed online synthesis 6 " method of-O-palmityl-Neohesperidin ester | |
| CN103667401B (en) | A kind of lipase-catalyzed online synthesis 6 " method of-O-lauroyl-Neohesperidin ester | |
| CN103667395B (en) | A kind of lipase-catalyzed online synthesis 6 " method of-O-palmityl-naringin dihydrochalcone ester | |
| CN103184250B (en) | Method for on-line synthesizing mannose-6-acetate by lipase catalysis | |
| CN103184254B (en) | Method of using lipase to catalyze and synthesize mannose-6-palmitate on line |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |