CN109593798A - A method of high-purity raphanin is produced with glucoraphenin - Google Patents
A method of high-purity raphanin is produced with glucoraphenin Download PDFInfo
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- CN109593798A CN109593798A CN201910029436.6A CN201910029436A CN109593798A CN 109593798 A CN109593798 A CN 109593798A CN 201910029436 A CN201910029436 A CN 201910029436A CN 109593798 A CN109593798 A CN 109593798A
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Abstract
The invention belongs to produce the technical field of raphanin, in particular to a kind of method with glucoraphenin production high-purity raphanin.The method of the present invention using glucoraphenin, myrosin, dialysis membrane tube as raw material, through the preparation of glucoraphenin reaction solution, reaction system preparation, hemodialysis reaction, reverse osmosis concentration, Centrifugical extraction and etc. production obtain raphanin.The method of the present invention introduces dialysis membrane tube and carries out hemodialysis reaction, realizes product and separates with the original position of substrate, enzyme, significantly improve glucoraphenin degradation rate, realizes the complete retention of myrosin, can be repeated for the production of raphanin.The features such as the method for the present invention has simple production process, stabilization, is easy to tissue amplification, and product yield is high, and equipment investment cost is low, and production cost is low, and comprehensive resource utilization rate is high, environmental-friendly.Have the characteristics that purity is high, no solvent residue, performance are stablized using the raphanin that the method for the present invention is prepared, can be widely applied to the industries such as medicine, food and daily use chemicals.
Description
Technical field
It is the invention belongs to produce the technical field of raphanin, in particular to a kind of to produce high-purity raphanin with glucoraphenin
Method.
Background technique
4- methylsulfinyl -3- cyclobutenyl sulphur glycosides, is commonly called as glucoraphenin (Glucoraphenin, C12H21NO10S3,
It 435.5Da), is a kind of sulphur glycosides of content at most in Radish seed.Myrosin (Myrosinase, EC 3.2.3.1) is also known as β-sulphur
For glucuroide, it is distributed mainly in root, stem, leaf, seedling and the seed of crucifer.Sulphur glycosides is urged myrosin
Under change effect, the fracture of S-glycosides key forms D-Glucose, hydrogen ion and unstable intermediate aglycone, and aglycone loses
It is reset after removing sulfate ion by Hoffmann or Lossen, generates biologically active isothiocyanates or salt.Radish
Glycosides is catalyzed through myrosin and produces 4- isothiocyanic acid base -1- (methylsulfinyl) -1- butylene, is commonly called as raphanin
(Sulforaphene, C6H9NOS2, 175.3Da), with bioactivity such as very strong anti-oxidant, anti-cell mutation, anticancers,
Application prospect is very wide.
The method of existing production raphanin, such as notification number of the October in 2017 of Granted publication on the 17th are 105198782 B of CN
The patent of invention of " extraction of raphanin and isolation and purification method in radish seed ", the method disclosed in the patent is: with radish kind
Son be raw material, freeze-dried, crushing, radish seed degreasing, enzymatic hydrolysis, extraction, concentration and etc. obtain raphanin crude extract, so
By making sheet, thin-layer chromatography and etc. be further purified to obtain raphanin.Major defect existing for this method has: 1. with radish kind
Son is that raw material carries out tissue production, and process route is long, process is complicated, is not easy to large-scale production；2. radish seed degreasing link,
Though helping to improve subsequent enzymolysis efficiency, need using a large amount of ether, higher cost, while causing part radish
The loss of glycosides reduces raphanin yield；3. using biggish compared to progress raphanin extraction in technique, when leading to subsequent concentration
Between it is long, raphanin is easily degraded, while it is higher that energy consumption is concentrated；4. isolating and purifying for raphanin is carried out using thin-layer chromatography, it is raw
Production capacity power is limited, is not suitable for industrialized production, and organic reagent consumption is more, seriously polluted.
Summary of the invention
The purpose of the present invention is the shortcomings for existing production raphanin method, provide a kind of high with glucoraphenin production
The method of purity raphanin, this method have simple production process, stabilization, are easy to tissue amplification, and product yield is high, equipment investment
The features such as at low cost, production cost is low, and comprehensive resource utilization rate is high, environmental-friendly.The radish prepared using the method for the present invention
Element has the characteristics that purity is high, no solvent residue, performance are stablized.
Mechanism of the invention is: myrosin and glucoraphenin are placed in the dialysis membrane that molecular cut off is 200Da or 250Da
In pipe, glucoraphenin-myrosin-dialysis membrane tube reaction system is constructed, then by glucoraphenin-myrosin-dialysis membrane tube reaction
System is placed in heat preservation chromatographic column, assembles dialysis membrane pipe reactor.Trailing plants in preference temperature and acid condition, reaction system
Glycosides is foretold by myrosin catalytic degradation, generates the primary products such as raphanin, glucose, sulfate radical.Due to myrosin molecular weight
Be 435.5Da for 65~75kDa, glucoraphenin molecular weight, be all larger than used membrane tube molecular cut off of dialysing, thus myrosin with
Glucoraphenin is by effectively catching in dialysis membrane tube.Meanwhile when by constantly purified water is pumped into dialysis membrane pipe reactor, generation
For the substances such as raphanin, glucose since molecular weight is less than dialysis membrane tube molecular cut off used, depressing in certain diffusion can not
Disconnected to flow slowly over dialysis membrane tube, dialysis enters in the purified water outside glucoraphenin-myrosin-dialysis membrane tube reaction system, makes to give birth to
Dialysis membrane pipe reactor is flowed continually out at object, thus isolated raphanin crude liquid.In reaction process, catalysate is constantly left
Glucoraphenin-myrosin-dialysis membrane tube reaction system realizes separation in situ, effectively reduces inhibition of the product to enzymic catalytic reaction
Effect, improves the degradation rate of glucoraphenin, also reduces the generation of secondary response, help to improve raphanin yield.It will dialysis
Membrane tube is introduced into reaction system, but also myrosin may be reused, effectively reduces production cost.Using reverse osmosis
Raphanin crude liquid is concentrated in concentration technique, and under a certain pressure, solvent and small-molecule substance can penetrate film, and raphanin
By effectively catching, to achieve the effect that concentration and purifying raphanin.Raphanin concentrate is carried out based on Centrifugical extraction technology
Extraction, centrifugal force are remarkably improved the separative efficiency of two-phase, reduce the water content in oily phase, improve the stabilization of raphanin
Property, and since n-hexane or ethyl acetate are high to the selectivity of raphanin, substantially increase raphanin purity.In addition, will be from
The heart vacuum concentration operating time extracted extends, and effectively improves the dissolvent residual of raphanin, increases Product Safety.
The object of the present invention is achieved like this: a method of with glucoraphenin produce high-purity raphanin, with glucoraphenin,
Myrosin, dialysis membrane tube be raw material, through the preparation of glucoraphenin reaction solution, reaction system preparation, hemodialysis reaction, reverse osmosis concentration,
Centrifugical extraction and etc. production obtain raphanin.Its specific method and step is as follows:
(1) glucoraphenin reaction solution is prepared
Using commercially available glucoraphenin as raw material, volume (mL) ratio according to quality (the g) ︰ purified water of glucoraphenin is 1 ︰'s 300~800
Glucoraphenin is added in purified water by ratio, then the sulfuric acid solution for being 5~10% with mass fraction adjusts pH to 3.0~4.5, stirs
After mixing uniformly, solution for standby, as glucoraphenin reaction solution are collected, is used as and handles in next step.
(2) glucoraphenin-myrosin-dialysis membrane tube reaction system is prepared
Using commercially available myrosin as raw material, the ratio that the mass ratio according to Hei mustard seed Mei ︰ glucoraphenin is 1 ︰ 1000~5000 will
Myrosin is slowly added into glucoraphenin reaction solution, and after mixing evenly, as glucoraphenin-myrosin reaction system is collected
It is spare.Using commercially available molecular cut off be 200Da or the dialysis membrane tube of 250Da is raw material, according to the appearance amount ︰ radish of dialysis membrane tube
Glycosides-myrosin reaction system volume ratio is the ratio of 1 ︰ 0.3~0.6, and glucoraphenin-myrosin reaction system is loaded on one
End has used reconfiguration film to press from both sides in closed dialysis membrane tube, then also closes the other end with reconfiguration film clamp, it is black just to prepare glucoraphenin-
Myrosase-dialysis membrane tube reaction system is used as and handles in next step.
(3) raphanin crude liquid is prepared
After the completion of (2) step, according to glucoraphenin-myrosin-dialysis membrane tube reaction for preparing of heat preservation chromatographic column ︰ (2) step
The volume ratio of system is the ratio of 1 ︰ 0.3~0.5, and glucoraphenin-myrosin-dialysis membrane tube reaction system is first placed in insulating layer
It analyses in column, then heats to 25~35 DEG C, just assemble dialysis membrane pipe reactor, it is spare.It is again 5~10% with mass fraction
Purified water is adjusted to pH identical with the glucoraphenin reaction solution of (1) step collection by sulfuric acid solution, collects acid purified water.Then
Acid purified water is pumped into dialysis membrane pipe reactor again, is dialysis membrane pipe reactor volume in the speed that is pumped into of acid purified water
1~3 times/it is small carry out continuous hemodialysis reaction at present, until in efflux without raphanin, respectively collect dialysis membrane tube it is anti-
Dialysis membrane pipe reactor after answering device efflux and reaction.For the dialysis membrane pipe reactor after the reaction of collection, contain black mustard
Sub- enzyme can continue to reuse after supplementing glucoraphenin according to the condition of (1) step；For the dialysis membrane pipe reactor outflow of collection
Liquid, as raphanin crude liquid are used as and handle in next step.
(4) raphanin concentrate is prepared
After the completion of (3) step, the raphanin crude liquid that (3) step is collected is pumped into reverse osmosis concentration device, in 0.08~0.15MPa
Under, reverse osmosis concentration is carried out, until the volume of reverse osmosis trapped fluid is reduced until the 1~5% of original volume, collects reverse osmosis respectively
Saturating permeate and reverse osmosis trapped fluid.For the reverse osmosis permeate of collection, after adjusting pH, it can be used as acid purified water and continue
It uses；For the reverse osmosis trapped fluid of collection, as raphanin concentrate, it is used as and handles in next step.
(5) high-purity raphanin is prepared
After the completion of (4) step, the volume ratio of the raphanin Nong Suo Ye ︰ ethyl acetate or n-hexane collected according to (4) step is 1 ︰ 2
The raphanin concentrate of collection and ethyl acetate or n-hexane are pumped into centrifugal extractor with cross-current flow, are flowed by~4 ratio
Speed is respectively 30~50mL/min, after Centrifugical extraction, collects water phase and oily phase respectively.For the water phase of collection, containing big
The substances such as glucose and micro raphanin are measured, after spray-dried, can be used as animal feed additive use.For the oil of collection
Phase is pumped into vacuum concentrator, under the conditions of vacuum degree is 0.08~0.095MPa, temperature is 30~40 DEG C, carries out vacuum
Concentration, stops after continuing concentration 3~6 hours when going out without evaporation liquid at drip, collects vacuum concentration evaporation liquid and vacuum respectively
Concentrate is concentrated.Liquid, predominantly ethyl acetate or n-hexane are evaporated for the vacuum concentration of collection, centrifugation extraction can be continued on for
It takes；For the vacuum concentration concentrate of collection, as high-purity raphanin.Glucoraphenin degradation rate is 92.7~95.1%, raphanin
Yield is 95.3~97.9%.Raphanin purity is 94.1~95.6%, no solvent residue, stores half a year at being -18 DEG C in temperature
It does not degrade.
The present invention is after adopting the above technical scheme, mainly have the following effects:
1, dialysis membrane tube is introduced in process of production and carry out hemodialysis reaction, realize product and separate with the original position of substrate, enzyme, effectively
Product inhibiton effect is reduced, glucoraphenin degradation rate is significantly improved.
2, the complete retention that myrosin in process of production, is realized using dialysis membrane tube, can be repeated for raphanin
Production, greatly reduce production cost.
3, process steps are few, are easy to tissue production, are at low cost, and dialysis membrane pipe reactor strong flexibility, can be real
Existing Flexible Production.
4, the use of Centrifugical extraction technology effectively improves the separative efficiency of water phase with oily phase, it is aqueous to reduce raphanin
Amount, improves product stability, and have the characteristics that treating capacity is big, high-efficient.
5, vacuum concentration duration in process of production, is properly increased, plays the role of reducing dissolvent residual, improves production
Product safety.
6, reverse osmosis to effectively increase production efficiency and raphanin purity with Centrifugical extraction technology, and have energy-saving
The advantages of.
The product prepared using the method for the present invention can be widely applied to the industries such as medicine, food and daily use chemicals.
Specific embodiment
With reference to embodiment, the present invention is further illustrated.
Embodiment 1
A method of high-purity raphanin being produced with glucoraphenin, steps are as follows for specific method:
(1) glucoraphenin reaction solution is prepared
Using commercially available glucoraphenin as raw material, the ratio that volume (mL) ratio according to quality (the g) ︰ purified water of glucoraphenin is 1 ︰ 300,
Glucoraphenin is added in purified water, then the sulfuric acid solution for being 5% with mass fraction adjusts pH to 3.0, after mixing evenly, collects
Solution for standby, as glucoraphenin reaction solution are used as and handle in next step.
(2) glucoraphenin-myrosin-dialysis membrane tube reaction system is prepared
Using commercially available myrosin as raw material, the ratio that the mass ratio according to Hei mustard seed Mei ︰ glucoraphenin is 1 ︰ 1000, by black mustard
Enzyme is slowly added into glucoraphenin reaction solution, and after mixing evenly, as glucoraphenin-myrosin reaction system, collection are spare.With
The dialysis membrane tube that commercially available molecular cut off is 200Da is raw material, is reacted according to appearance amount ︰ glucoraphenin-myrosin of dialysis membrane tube
The volume ratio of system is the ratio of 1 ︰ 0.3, has used reconfiguration film folder closed loaded on one end glucoraphenin-myrosin reaction system
In membrane tube of dialysing, then the other end is also closed with reconfiguration film clamp, just prepares glucoraphenin-myrosin-dialysis membrane tube reaction
System is used as and handles in next step.
(3) raphanin crude liquid is prepared
After the completion of (2) step, according to glucoraphenin-myrosin-dialysis membrane tube reaction for preparing of heat preservation chromatographic column ︰ (2) step
The volume ratio of system is the ratio of 1 ︰ 0.3, and glucoraphenin-myrosin-dialysis membrane tube reaction system is first placed in heat preservation chromatographic column
In, 25 DEG C are then heated to, dialysis membrane pipe reactor is just assembled, it is spare.It again will be pure with the sulfuric acid solution that mass fraction is 5%
Change water and be adjusted to pH identical with the glucoraphenin reaction solution of (1) step collection, collects acid purified water.Then acidity is purified again
Water is pumped into dialysis membrane pipe reactor, is pumped into 1 times/hour that speed is dialysis membrane pipe reactor volume in acid purified water
Under, continuous hemodialysis reaction is carried out, until in efflux without raphanin, collects dialysis membrane pipe reactor efflux and anti-respectively
Dialysis membrane pipe reactor after answering.For the dialysis membrane pipe reactor after the reaction of collection, containing myrosin, according to (1)
It can continue to reuse after the condition supplement glucoraphenin of step；For the dialysis membrane pipe reactor efflux of collection, as raphanin
Crude liquid is used as and handles in next step.
(4) raphanin concentrate is prepared
After the completion of (3) step, the raphanin crude liquid that (3) step is collected is pumped into reverse osmosis concentration device, at 0.08MPa, into
Row reverse osmosis concentration collects reverse osmosis permeate until the volume of reverse osmosis trapped fluid is reduced until the 1% of original volume respectively
With reverse osmosis trapped fluid.For the reverse osmosis permeate of collection, after adjusting pH, it can be used as acid purified water and continue to use；It is right
In the reverse osmosis trapped fluid of collection, as raphanin concentrate, it is used as and handles in next step.
(5) high-purity raphanin is prepared
After the completion of (4) step, the volume ratio for the raphanin Nong Suo Ye ︰ ethyl acetate collected according to (4) step is the ratio of 1 ︰ 2,
The raphanin concentrate and ethyl acetate of collection are pumped into centrifugal extractor with cross-current flow, flow velocity is respectively 30mL/min,
After Centrifugical extraction, water phase and oily phase are collected respectively.For the water phase of collection, contain a large amount of glucose and micro raphanin etc.
Substance after spray-dried, can be used as animal feed additive use.For the oily phase of collection, it is pumped into vacuum concentrator,
It under the conditions of vacuum degree is 0.08MPa, temperature is 30 DEG C, is concentrated in vacuo, continues concentration 3 when going out without evaporation liquid at drip
Stop after hour, collects vacuum concentration evaporation liquid and vacuum concentration concentrate respectively.Liquid is evaporated for the vacuum concentration of collection, it is main
To be ethyl acetate, Centrifugical extraction can be continued on for；For the vacuum concentration concentrate of collection, as high-purity raphanin.Trailing plants
Foretelling glycosides degradation rate is 92.7%, and raphanin yield is 95.3%.Raphanin purity is 94.1%, no solvent residue, is -18 in temperature
Storage half a year does not degrade at DEG C.
Embodiment 2
A method of high-purity raphanin being produced with glucoraphenin, steps are as follows for specific method:
(1) glucoraphenin reaction solution is prepared
Using commercially available glucoraphenin as raw material, the ratio that volume (mL) ratio according to quality (the g) ︰ purified water of glucoraphenin is 1 ︰ 500,
Glucoraphenin is added in purified water, then the sulfuric acid solution for being 8% with mass fraction adjusts pH to 4.0, after mixing evenly, collects
Solution for standby, as glucoraphenin reaction solution are used as and handle in next step.
(2) glucoraphenin-myrosin-dialysis membrane tube reaction system is prepared
Using commercially available myrosin as raw material, the ratio that the mass ratio according to Hei mustard seed Mei ︰ glucoraphenin is 1 ︰ 3000, by black mustard
Enzyme is slowly added into glucoraphenin reaction solution, and after mixing evenly, as glucoraphenin-myrosin reaction system, collection are spare.With
The dialysis membrane tube that commercially available molecular cut off is 250Da is raw material, is reacted according to appearance amount ︰ glucoraphenin-myrosin of dialysis membrane tube
The volume ratio of system is the ratio of 1 ︰ 0.4, has used reconfiguration film folder closed loaded on one end glucoraphenin-myrosin reaction system
In membrane tube of dialysing, then the other end is also closed with reconfiguration film clamp, just prepares glucoraphenin-myrosin-dialysis membrane tube reaction
System is used as and handles in next step.
(3) raphanin crude liquid is prepared
After the completion of (2) step, according to glucoraphenin-myrosin-dialysis membrane tube reaction for preparing of heat preservation chromatographic column ︰ (2) step
The volume ratio of system is the ratio of 1 ︰ 0.4, and glucoraphenin-myrosin-dialysis membrane tube reaction system is first placed in heat preservation chromatographic column
In, 30 DEG C are then heated to, dialysis membrane pipe reactor is just assembled, it is spare.It again will be pure with the sulfuric acid solution that mass fraction is 8%
Change water and be adjusted to pH identical with the glucoraphenin reaction solution of (1) step collection, collects acid purified water.Then acidity is purified again
Water is pumped into dialysis membrane pipe reactor, is pumped into 2 times/hour that speed is dialysis membrane pipe reactor volume in acid purified water
Under, continuous hemodialysis reaction is carried out, until in efflux without raphanin, collects dialysis membrane pipe reactor efflux and anti-respectively
Dialysis membrane pipe reactor after answering.For the dialysis membrane pipe reactor after the reaction of collection, containing myrosin, according to (1)
It can continue to reuse after the condition supplement glucoraphenin of step；For the dialysis membrane pipe reactor efflux of collection, as raphanin
Crude liquid is used as and handles in next step.
(4) raphanin concentrate is prepared
After the completion of (3) step, the raphanin crude liquid that (3) step is collected is pumped into reverse osmosis concentration device, at 0.11MPa, into
Row reverse osmosis concentration collects reverse osmosis permeate until the volume of reverse osmosis trapped fluid is reduced until the 3% of original volume respectively
With reverse osmosis trapped fluid.For the reverse osmosis permeate of collection, after adjusting pH, it can be used as acid purified water and continue to use；It is right
In the reverse osmosis trapped fluid of collection, as raphanin concentrate, it is used as and handles in next step.
(5) high-purity raphanin is prepared
After the completion of (4) step, the volume ratio for the raphanin Nong Suo Ye ︰ n-hexane collected according to (4) step is the ratio of 1 ︰ 3, will
The raphanin concentrate and n-hexane of collection are pumped into centrifugal extractor with cross-current flow, and flow velocity is respectively 40mL/min, centrifugation
After extraction, water phase and oily phase are collected respectively.For the water phase of collection, contain the objects such as a large amount of glucose and micro raphanin
Matter after spray-dried, can be used as animal feed additive use.For the oily phase of collection, it is pumped into vacuum concentrator,
It under the conditions of vacuum degree is 0.09MPa, temperature is 35 DEG C, is concentrated in vacuo, it is small to continue concentration 4 when going out without evaporation liquid at drip
When after stop, respectively collect vacuum concentration evaporation liquid and vacuum concentration concentrate.Liquid is evaporated for the vacuum concentration of collection, mainly
For n-hexane, Centrifugical extraction can be continued on for；For the vacuum concentration concentrate of collection, as high-purity raphanin.Glucoraphenin
Degradation rate is 94.2%, and raphanin yield is 96.8%.Raphanin purity is 95.1%, no solvent residue, in the case where temperature is -18 DEG C
Storage half a year does not degrade.
Embodiment 3
A method of high-purity raphanin being produced with glucoraphenin, steps are as follows for specific method:
(1) glucoraphenin reaction solution is prepared
Using commercially available glucoraphenin as raw material, the ratio that volume (mL) ratio according to quality (the g) ︰ purified water of glucoraphenin is 1 ︰ 800,
Glucoraphenin is added in purified water, then the sulfuric acid solution for being 10% with mass fraction adjusts pH to 4.5, after mixing evenly, collects
Solution for standby, as glucoraphenin reaction solution are used as and handle in next step.
(2) glucoraphenin-myrosin-dialysis membrane tube reaction system is prepared
Using commercially available myrosin as raw material, the ratio that the mass ratio according to Hei mustard seed Mei ︰ glucoraphenin is 1 ︰ 5000, by black mustard
Enzyme is slowly added into glucoraphenin reaction solution, and after mixing evenly, as glucoraphenin-myrosin reaction system, collection are spare.With
The dialysis membrane tube that commercially available molecular cut off is 200Da is raw material, is reacted according to appearance amount ︰ glucoraphenin-myrosin of dialysis membrane tube
The volume ratio of system is the ratio of 1 ︰ 0.6, has used reconfiguration film folder closed loaded on one end glucoraphenin-myrosin reaction system
In membrane tube of dialysing, then the other end is also closed with reconfiguration film clamp, just prepares glucoraphenin-myrosin-dialysis membrane tube reaction
System is used as and handles in next step.
(3) raphanin crude liquid is prepared
After the completion of (2) step, according to glucoraphenin-myrosin-dialysis membrane tube reaction for preparing of heat preservation chromatographic column ︰ (2) step
The volume ratio of system is the ratio of 1 ︰ 0.5, and glucoraphenin-myrosin-dialysis membrane tube reaction system is first placed in heat preservation chromatographic column
In, 35 DEG C are then heated to, dialysis membrane pipe reactor is just assembled, it is spare.It again will be pure with the sulfuric acid solution that mass fraction is 10%
Change water and be adjusted to pH identical with the glucoraphenin reaction solution of (1) step collection, collects acid purified water.Then acidity is purified again
Water is pumped into dialysis membrane pipe reactor, is pumped into 3 times/hour that speed is dialysis membrane pipe reactor volume in acid purified water
Under, continuous hemodialysis reaction is carried out, until in efflux without raphanin, collects dialysis membrane pipe reactor efflux and anti-respectively
Dialysis membrane pipe reactor after answering.For the dialysis membrane pipe reactor after the reaction of collection, containing myrosin, according to (1)
It can continue to reuse after the condition supplement glucoraphenin of step；For the dialysis membrane pipe reactor efflux of collection, as raphanin
Crude liquid is used as and handles in next step.
(4) raphanin concentrate is prepared
After the completion of (3) step, the raphanin crude liquid that (3) step is collected is pumped into reverse osmosis concentration device, at 0.15MPa, into
Row reverse osmosis concentration collects reverse osmosis permeate until the volume of reverse osmosis trapped fluid is reduced until the 5% of original volume respectively
With reverse osmosis trapped fluid.For the reverse osmosis permeate of collection, after adjusting pH, it can be used as acid purified water and continue to use；It is right
In the reverse osmosis trapped fluid of collection, as raphanin concentrate, it is used as and handles in next step.
(5) high-purity raphanin is prepared
After the completion of (4) step, the volume ratio for the raphanin Nong Suo Ye ︰ ethyl acetate collected according to (4) step is the ratio of 1 ︰ 4,
The raphanin concentrate and ethyl acetate of collection are pumped into centrifugal extractor with cross-current flow, flow velocity is respectively 50mL/min,
After Centrifugical extraction, water phase and oily phase are collected respectively.For the water phase of collection, contain a large amount of glucose and micro raphanin etc.
Substance after spray-dried, can be used as animal feed additive use.For the oily phase of collection, it is pumped into vacuum concentrator,
It under the conditions of vacuum degree is 0.095MPa, temperature is 40 DEG C, is concentrated in vacuo, continues to be concentrated when going out without evaporation liquid at drip
Stop after 6 hours, collects vacuum concentration evaporation liquid and vacuum concentration concentrate respectively.Liquid is evaporated for the vacuum concentration of collection,
Predominantly ethyl acetate can continue on for Centrifugical extraction；For the vacuum concentration concentrate of collection, as high-purity raphanin.
Glucoraphenin degradation rate is 95.1%, and raphanin yield is 97.9%.Raphanin purity be 95.6%, no solvent residue, temperature be-
Storage half a year does not degrade at 18 DEG C.
Claims (1)
1. a kind of method with glucoraphenin production high-purity raphanin, it is characterised in that steps are as follows for specific method:
(1) glucoraphenin reaction solution is prepared
It is the ratio of 1 g ︰, 300~800 mL according to the volume ratio of the Zhi Liang ︰ purified water of glucoraphenin using commercially available glucoraphenin as raw material
Example, glucoraphenin is added in purified water, then the sulfuric acid solution for being 5~10% with mass fraction adjusts pH to 3.0~4.5, stirring
After uniformly, solution for standby, as glucoraphenin reaction solution are collected, is used as and handles in next step；
(2) glucoraphenin-myrosin-dialysis membrane tube reaction system is prepared
Using commercially available myrosin as raw material, the ratio that the mass ratio according to Hei mustard seed Mei ︰ glucoraphenin is 1 ︰ 1000~5000 will
Myrosin is slowly added into glucoraphenin reaction solution, and after mixing evenly, as glucoraphenin-myrosin reaction system is collected
It is spare；Using commercially available molecular cut off be 200Da or the dialysis membrane tube of 250Da is raw material, according to the appearance amount ︰ radish of dialysis membrane tube
Glycosides-myrosin reaction system volume ratio is the ratio of 1 ︰ 0.3~0.6, and glucoraphenin-myrosin reaction system is loaded on one
End has used reconfiguration film to press from both sides in closed dialysis membrane tube, then also closes the other end with reconfiguration film clamp, it is black just to prepare glucoraphenin-
Myrosase-dialysis membrane tube reaction system is used as and handles in next step；
(3) raphanin crude liquid is prepared
After the completion of (2) step, according to glucoraphenin-myrosin-dialysis membrane tube reaction for preparing of heat preservation chromatographic column ︰ (2) step
The volume ratio of system is the ratio of 1 ︰ 0.3~0.5, and glucoraphenin-myrosin-dialysis membrane tube reaction system is first placed in insulating layer
It analyses in column, then heats to 25~35 DEG C, just assemble dialysis membrane pipe reactor, it is spare；It is again 5~10% with mass fraction
Purified water is adjusted to pH identical with the glucoraphenin reaction solution of (1) step collection by sulfuric acid solution, collects acid purified water；Then
Acid purified water is pumped into dialysis membrane pipe reactor again, is dialysis membrane pipe reactor volume in the speed that is pumped into of acid purified water
1~3 times/it is small carry out continuous hemodialysis reaction at present, until in efflux without raphanin, respectively collect dialysis membrane tube it is anti-
Dialysis membrane pipe reactor after answering device efflux and reaction；For the dialysis membrane pipe reactor efflux of collection, as raphanin
Crude liquid is used as and handles in next step；
(4) raphanin concentrate is prepared
After the completion of (3) step, the raphanin crude liquid that (3) step is collected is pumped into reverse osmosis concentration device, in 0.08~0.15MPa
Under, reverse osmosis concentration is carried out, until the volume of reverse osmosis trapped fluid is reduced until the 1~5% of original volume, collects reverse osmosis respectively
Saturating permeate and reverse osmosis trapped fluid；For the reverse osmosis trapped fluid of collection, as raphanin concentrate, it is used as in next step
Reason；
(5) high-purity raphanin is prepared
After the completion of (4) step, the volume ratio of the raphanin Nong Suo Ye ︰ ethyl acetate or n-hexane collected according to (4) step is 1 ︰ 2
The raphanin concentrate of collection and ethyl acetate or n-hexane are pumped into centrifugal extractor with cross-current flow, are flowed by~4 ratio
Speed is respectively 30~50mL/min, after Centrifugical extraction, collects water phase and oily phase respectively；For the oily phase of collection, it is pumped into
In vacuum concentrator, under the conditions of vacuum degree is 0.08~0.095MPa, temperature is 30~40 DEG C, it is concentrated in vacuo, to nothing
Evaporation liquid stops after continuing concentration 3~6 hours when going out at drip, collects vacuum concentration evaporation liquid and vacuum concentration concentration respectively
Liquid；For the vacuum concentration concentrate of collection, as high-purity raphanin；Glucoraphenin degradation rate is 92.7~95.1%, raphanin
Yield is 95.3~97.9%；Raphanin purity is 94.1~95.6%, no solvent residue, stores half a year at being -18 DEG C in temperature
It does not degrade.
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