CN103667393B - A kind of lipase-catalyzed online synthesis 6 " method of-O-palmityl-neohesperidin dihydrochalcone ester - Google Patents
A kind of lipase-catalyzed online synthesis 6 " method of-O-palmityl-neohesperidin dihydrochalcone ester Download PDFInfo
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Abstract
The invention discloses a kind of method of lipase-catalyzed online synthesis 6''-O-palmityl-neohesperidin dihydrochalcone ester, described method is: take mol ratio as neohesperidin dihydrochalcone and the vinyl palmitate of 1:1 ~ 12 be raw material, with 0.5 ~ 1.0g Lipozyme RMIM for catalyzer, with the mixed solvent of tertiary amyl alcohol and DMSO for reaction solvent, 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, 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''-O-palmityl-neohesperidin dihydrochalcone ester through conventional aftertreatment.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 method of lipase-catalyzed online controlled selectivity synthesis 6''-O-palmityl-neohesperidin dihydrochalcone ester.
(2) background technology
Neohesperidin dihydrochalcone (Neohesperidin dihydrochalcone) is the neohesperidin that separation and Extraction from natural citrus class plant the obtained flavone derivative through peroxidation, its sugariness is 500 ~ 700 times of sucrose, it has high sugariness, the feature such as low in calories, nontoxic, safe, there is anticancer, antibacterial, the multiple biological activity such as antioxygen, decreasing cholesterol, be therefore applied to food, medicine and daily chemical industry.In addition, neohesperidin dihydrochalcone clean taste, with special delicate fragrance, have the effect of splendid shielding bitter taste, therefore neohesperidin dihydrochalcone is specially adapted to vegetable based food processing, jam, chewing gum, toothpaste and sucks in tablet.
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 dihydrochalcone is as applying one of maximum flavonoid compound, and the neohesperidin dihydrochalcone fatty acid ester researching and developing its physiologically active stronger will have a good application prospect.
At present, the route of synthesis of neohesperidin dihydrochalcone fatty acid ester mainly carries out enzymatic reaction in organic medium, carries out acylations to the hydroxyl on glycosyl, specifically comprises acidylate, esterification and transesterification reaction three kinds of methods.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 dihydrochalcone enzymatic selective esterification new technology is found significant for the production of development neohesperidin dihydrochalcone 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, there is the enzymatic regioselectivity esterification of more Chinese scholars to flavonoid compound in organic medium to be studied, but also do not have for the research of the enzymatic modification of neohesperidin dihydrochalcone.And the enzymatic regioselectivity esterification process of flavonoid compound often needs the longer reaction times (24h) in the organic medium reported, 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 dihydrochalcone is the neohesperidin that separation and Extraction from natural citrus class plant the obtained flavone derivative through peroxidation, 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 dihydrochalcone is modified and not only can improve bioavailability, its application in fields such as medicine, food can also be expanded.And neohesperidin dihydrochalcone is the sugary flavone derivative of a class, we can utilize the enzymatic regioselectivity esterification of Research foundation to neohesperidin dihydrochalcone in early stage to study.Therefore we have studied the method for lipase-catalyzed selectivity synthesis 6''-O-palmityl-neohesperidin dihydrochalcone ester in microfluidic channel reactor, be intended to the online controlled method for selective synthesis of the neohesperidin dihydrochalcone palmitinic acid monoesters finding a kind of high-efficiency environment friendly.Due to the neohesperidin dihydrochalcone monoesters that different chain length replaces, its solvability, cell permeabilities etc. have a great difference, thus microflow control technique is utilized to synthesize the neohesperidin dihydrochalcone monoester class compound of different chain length replacement, its bioavailability can be furtherd investigate, expand its application in medicine and the field such as food.
(3) summary of the invention
The technical problem to be solved in the present invention is to provide the novel process of lipase-catalyzed selectivity synthesis 6''-O-palmityl-neohesperidin dihydrochalcone ester in a kind of microfluidic channel reactor, 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 method of lipase-catalyzed online synthesis 6''-O-palmityl-neohesperidin dihydrochalcone 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 dihydrochalcone being 1:1 ~ 1:12 with the ratio of amount of substance and vinyl palmitate 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 obtains 6''-O-palmityl-neohesperidin dihydrochalcone ester through 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 dihydrochalcone with the mixed solvent (tertiary amyl alcohol: DMSO=4:1) of tertiary amyl alcohol and DMSO, as long as its consumption ensures that neohesperidin dihydrochalcone can fully be dissolved, and is loaded in syringe for subsequent use; Only dissolve vinyl palmitate 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 dihydrochalcone 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 palmitate 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 dihydrochalcone and the mixing solutions of DMSO and be dissolved with the tert-amyl alcohol solution of vinyl palmitate time, volumetric usage for dissolving the tertiary amyl alcohol of vinyl palmitate is preferably equal with the volumetric usage of dissolving the tertiary amyl alcohol of neohesperidin dihydrochalcone 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 dihydrochalcone counts 10 ~ 25mL/mmol with the amount of substance of neohesperidin dihydrochalcone 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 Rhizomucor miehei, is produce through submerged fermentation with a kind of gene-modified aspergillus oryzae (Aspergillus oryzae) microorganism.Lipozyme RM IM 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 dihydrochalcone and vinyl palmitate is preferably 1:8 ~ 12, 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, and gained reaction solution can obtain 6''-O-palmityl-neohesperidin dihydrochalcone ester by conventional post-treating method.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-palmityl-neohesperidin dihydrochalcone 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 dihydrochalcone, but lipase-catalyzed selectivity synthesis 6''-O-palmityl-neohesperidin dihydrochalcone ester has higher transformation efficiency and selectivity 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
1h NMR confirms.This shows that the enzymatic selectivity synthesis of neohesperidin dihydrochalcone cetylate 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 dihydrochalcone-6''-O-cetylate in microfluidic channel reactor of the present invention, 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 an interface.
The synthesis of embodiment 1:6''-O-palmityl-neohesperidin dihydrochalcone ester
Equipment therefor is with reference to figure 1.Neohesperidin dihydrochalcone (0.49mmol) is dissolved in 10mL tertiary amyl alcohol: in the mixed solvent of DMSO=4:1 (v/v), vinyl palmitate (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, TLC follows the tracks of wash-out process simultaneously, the elutriant containing single product obtained is merged evaporate to dryness, obtains flaxen monoesters crystal, obtains neohesperidin dihydrochalcone-6''-cetylate, is 6''-O-palmityl-neohesperidin dihydrochalcone ester.The selectivity (the neohesperidin dihydrochalcone ester total content of the content/generation of neohesperidin dihydrochalcone-6''-cetylate) that HPLC detects neohesperidin dihydrochalcone transformation efficiency 84%, neohesperidin dihydrochalcone-6''-cetylate is 100%.
Nuclear-magnetism characterization result is as follows:
1h-NMR (DMSO-d6, δ, ppm): neohesperidin dihydrochalcone-6''-cetylate: 12.33 (s, 2H ,-OH
5), 9.15 (s ,-OH
5'), 7.03 (d, 2H, J=8.4Hz, H
2'and H
6'), 6.67 (d, 2H, J=8.5Hz, H
3'and H
5'), 5.99 (s, 2H, H
6and H
8), 5.44 (m, 1H, H
1''), 5.41 (m, 1H, 2'''-OH of neohesperidindihydrochalcone), 5.12 (m, 1H, H1'''), 5.08 (d, 1H, J=5.1Hz, 4'''-OH of neohesperidindihydrochalcone), 4.70 (d, 1H, J=4.8Hz, 4''-OH of neohesperidin dihydrochalcone), 4.68 (d, 1H, J=4.4Hz, 3''-OH of neohesperidin dihydrochalcone), 4.50 (d, 1H, J=5.8Hz, 3'''-OH of neohesperidin dihydrochalcone), 4.26 (d, 1H, J=11.9Hz, H
6''acylated), 4.04 (m, 1H, H
6''acylated), 3.73-3.16 (8H, H of rhamnoglucosyl), 3.22 (m, 2H, H
3), 2.78 (t, 2H, J=7.7Hz, H
2), 2.27 (m, 2H ,-CH
2-CO-), 1.20 (m, 3H of CH
3ofrhamnosyl), 1.26 (m, 26H of (CH
2)
13of palmitoyl), 0.84 (t, 3H, J=7.0Hz, CH
3).
IR(KBr,cm
-1):3415(OH),1740(C=O),1643(C=C).
ESI-MS(m/z):886(M
1+2H
2O-H)
-,M
1corresponding exactly to the molecular weightof6''-O-palmitoyl-neohesperidin dihydrochalcone.
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 dihydrochalcone cetylate be 52 DEG C.
Embodiment 6-10
The substrate mol ratio changing vinyl palmitate and neohesperidin dihydrochalcone 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 dihydrochalcone is that 0.49mmol is constant, changes the consumption of vinyl palmitate.Other are with embodiment 1, and result is as shown in table 2.
Table 2: the impact that neohesperidin dihydrochalcone and the comparison of vinyl palmitate substrate are reacted
The result of table 2 shows, along with the increase of reactant vinyl palmitate, 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 dihydrochalcone has substantially transformed completely in order to neohesperidin dihydrochalcone-6''-cetylate.If now continue the consumption increasing reactant vinyl palmitate, the transformation efficiency of reaction and selectivity will be caused to reduce, thus, the best substrate of this reaction is than being 8:1, under the reaction conditions, neohesperidin dihydrochalcone transforms substantially completely in order to neohesperidin dihydrochalcone-6''-cetylate.
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 | 44 | 100 |
| 12 | 25 | 60 | 100 |
| 13 | 30 | 72 | 100 |
| 1 | 35 | 84 | 100 |
| 14 | 40 | 76 | 99 |
The result of table 3 shows, neohesperidin dihydrochalcone-6''-palmitinic acid the monoesters that 20min can obtain 44% is carried out in reaction, selectivity is now 100%, and neohesperidin dihydrochalcone is be converted into neohesperidin dihydrochalcone-6''-palmitinic acid monoesters completely substantially.Along with the increase in reaction times, the transformation efficiency of reaction increases gradually, when 35min is carried out in reaction, and the transformation efficiency of reaction and selectivity optimum, neohesperidin dihydrochalcone-6''-palmitinic acid monoesters transformation efficiency can reach 84%, 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, in microfluidic channel reactor, the Best Times of neohesperidin dihydrochalcone-6''-palmitinic acid monoesters synthesis is 35min.
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 | 22 | 100 |
| Comparative example 3 | Bacillus alkaline protease | 6 | 100 |
| Embodiment 1 | Lipozyme RMIM | 84 | 100 |
The result of table 4 shows, for the regioselectivity esterification of enzymatic neohesperidin dihydrochalcone in microfluidic channel reactor, different enzymes has fairly obvious impact to reaction.Utilize Lipozyme TLIM catalyzed reaction, the transformation efficiency of neohesperidin dihydrochalcone-6''-palmitinic acid monoesters is 50%.And utilizing this reaction of bacillus alkaline protease catalysis, the transformation efficiency of neohesperidin dihydrochalcone-6''-palmitinic acid monoesters is only 6%.From the result of table 4, for the regioselectivity esterification of enzymatic neohesperidin dihydrochalcone in microfluidic channel reactor, the most effective catalyzer is Lipozyme RMIM, the transformation efficiency of neohesperidin dihydrochalcone-6''-palmitinic acid monoesters is 84%, and selectivity is 100%.
Comparative example 4 ~ 5
Change different types of flavonoid compound in micro-fluidic micro passage reaction, change the neohesperidin dihydrochalcone (0.49mmol) of embodiment 1 into hesperidine 0.49mmol(comparative example 4 respectively), saligenin 0.49mmol(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 | 3 | 100 |
| Comparative example 5 | Saligenin | 12 | 100 |
| Embodiment 1 | Neohesperidin dihydrochalcone | 84 | 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 3% to hesperidine, and reaction and difficulty thereof, react hardly.The reaction result of saligenin is also undesirable, and transformation efficiency is only 12%.From the result of table 5, for the regioselectivity esterification of enzymatic flavonoid compound in microfluidic channel reactor, neohesperidin dihydrochalcone can obtain more satisfactory reaction result, and the transformation efficiency of reaction can reach 84%, and selectivity is 100%.
Claims (6)
1. a lipase-catalyzed online synthesis 6 " method of-O-palmityl-neohesperidin dihydrochalcone 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 dihydrochalcone being 1:8 ~ 12 with the ratio of amount of substance and vinyl palmitate 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 aftertreatment obtained 6 "-O-palmityl-neohesperidin dihydrochalcone ester.
2. lipase-catalyzed online synthesis 6 as claimed in claim 1 " method of-O-palmityl-neohesperidin dihydrochalcone ester; it is characterized in that: described method comprises the following steps: to be first that the tertiary amyl alcohol of 4:1 and the mixed solvent of DMSO dissolve neohesperidin dihydrochalcone by volume ratio, is loaded in syringe for subsequent use; Dissolve vinyl palmitate 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. lipase-catalyzed online synthesis 6 as claimed in claim 1 " method of-O-palmityl-neohesperidin dihydrochalcone ester, it is characterized in that: described microfluidic channel reactor comprises thermostat container, and described reaction channel is placed in thermostat container.
4. lipase-catalyzed online synthesis 6 as claimed in claim 2 " method of-O-palmityl-neohesperidin dihydrochalcone ester, it is characterized in that: described microfluidic channel reactor comprises thermostat container, and described reaction channel is placed in thermostat container.
5. lipase-catalyzed online synthesis 6 as claimed in claim 1 " method of-O-palmityl-neohesperidin dihydrochalcone ester, it is characterized in that: described neohesperidin dihydrochalcone is 1:8 with the ratio of the amount of substance of vinyl palmitate.
6. lipase-catalyzed online synthesis 6 as claimed in claim 1 " method of-O-palmityl-neohesperidin dihydrochalcone ester, it is characterized in that: described acylation reaction temperature is 52 DEG C, the described acylation reaction time is 35min.
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| CN103184249A (en) * | 2011-12-31 | 2013-07-03 | 浙江工业大学 | Method for on-line synthesizing glucose-6-palmitate by lipase catalysis |
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| CN103184249A (en) * | 2011-12-31 | 2013-07-03 | 浙江工业大学 | Method for on-line synthesizing glucose-6-palmitate by lipase catalysis |
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| Regioselective acylation of flavonoids catalyzed by lipase in low toxicity media;Athina Kontogianni et al.;《Eur. J. Lipid Sci. Technol.》;20011031;第103卷(第10期);第655–660页 * |
| 黄建蓉.磁场影响有机介质中酶促合成柚皮苷酯机理及其特性研究.《中国博士学位论文全文数据库,工程科技I辑》.2010,第1.1.3.3、1.2、2.2.3.5节. * |
| 黄酮类化合物的分子修饰;李卫等;《食品科学》;20050815;第25卷(第8期);505-507 * |
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