CN102424662A - Separation and purification method for vitamin A palmitate - Google Patents
Separation and purification method for vitamin A palmitate Download PDFInfo
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Abstract
The present invention discloses a separation and purification method for vitamin A palmitate. The method is applicable for separation of a reaction product from the following reaction: retinol acetate reacts with palmitic acid by the catalysis effect of immobilized lipase to synthesize the vitamin A palmitate. The separation and purification method comprises the following steps: 1) removing the palmitic acid by low temperature crystallization; 2) separating the vitamin A palmitate and the retinol acetate by a liquid-liquid extraction and separation method or a silica gel column chromatography separation method. According to the present invention, the process conditions of the separation and the purification are optimized; the liquid-liquid extraction and separation method of the present invention has advantages of simple operation and low equipment requirements; with the silica gel column chromatography separation method, the high purity vitamin A palmitate can be obtained.
Description
Technical field
The invention belongs to enzyme process synthesise vitamins A cetylate field, be specifically related to the separation purification method in the enzyme process synthesise vitamins A cetylate process.
Background technology
The vitamin A cetylate is one of topmost series product of vitamin A; Be widely used in food, medicine, dietary supplements, makeup, fodder additives etc.; It is the indispensable a kind of material of human homergy; It can keep and promote people's bulk-growth, growth, reproduction and cell membrane stability, to the tangible effect of being formed with of vision, and; The vitamin A cetylate also helps the breeding of cell and distinguishes non-viable non-apoptotic cell, and can produce the combined with radical of transgenation with those as a kind of inhibitor.The required vitamin A cetylate of human homergy mainly is to be formed by esterification on small bowel by vitamin A usually; Play a role again after being transported in the liver storage by lymph again, so the vitamin A cetylate has vital role to the regular intracellular metabolite of human body.Vitamin A, vitamin A acetic ester, vitamin A palm ester are that China can allow the vitamin A series additive that adds, but vitamin A, vitamin A acetic ester are unstable to heat, auroral poles.The relative vitamin A of vitamin A palm ester, vitamin A acetic ester chemical property are stablized, and are difficult for advantages such as decomposition, have been widely used in makeup, medicine, feed etc.
Vitamin A Palmitate 1.7 M.I.U/Gram can adopt fixed lipase catalyzed synthetic.The reaction mechanism of fixed lipase catalyzed synthesise vitamins A cetylate is following:
Enzymic catalytic reaction is generally reversible reaction, and can draw thus has intact and the palmitinic acid of unreacted still in the reaction afterreaction liquid, and product Vitamin A Palmitate 1.7 M.I.U/Gram and acetate also have the reaction solvent sherwood oil.
The aftertreatment of enzyme process synthesise vitamins A cetylate mainly comprises two kinds of strategies; A kind of is through optimal conditions; Improve the conversion of, make transformation efficiency, so just need not carry out the separation and purification operation as far as possible near 100% to Vitamin A Palmitate 1.7 M.I.U/Gram; And direct crystallization just can obtain the very high Vitamin A Palmitate 1.7 M.I.U/Gram of purity, and this also is a strategy of producing Davitin A at present as vitamin A manufacturing enterprises of a few family such as Xiamen Jin Dawei; Another kind is not require that transformation efficiency is very high, after reaction finishes, carries out separation and purification, obtains the very high Vitamin A Palmitate 1.7 M.I.U/Gram of purity through one or several lock out operation.The height of separation and purification cost is the key whether this synthesis technique can industrialization, therefore, considers from the industrialization economy principle, requires the separation and purification operation simple as far as possible, to reduce production costs, increases enterprise profit.
Summary of the invention
To the deficiency of prior art, the invention provides a kind of separation purification method of Vitamin A Palmitate 1.7 M.I.U/Gram, thereby can obtain the very high Vitamin A Palmitate 1.7 M.I.U/Gram finished product of purity.
For achieving the above object, the present invention realizes through following technical scheme:
The separation purification method of Vitamin A Palmitate 1.7 M.I.U/Gram comprises the steps:
1) removal of palmitinic acid: earlier with and palmitinic acid through the reaction product of fixed lipase catalyzed synthesise vitamins A cetylate at 4 ℃ of condition held 20 h, separate out the palmitinic acid crystallization, remove by filter the palmitinic acid crystallization; Supernatant liquid after then will filtering is placed on 8 h under-20 ℃ of conditions, separates out the palmitinic acid crystallization again, removes by filter the palmitinic acid crystallization once more, obtains filtered liq;
2) separate Vitamin A Palmitate 1.7 M.I.U/Gram and: the filtered liq that obtains separates Vitamin A Palmitate 1.7 M.I.U/Gram and with liquid-liquid extraction partition method or silica gel column chromatography partition method.
Step 2) the liquid-liquid extraction partition method in is:
With the filtered liq that obtains with the mixed solvent of ethanol-water solution and sherwood oil as the extraction solvent extracting and separating, extraction temperature is 4 ℃.
In the mixed solvent of ethanol-water solution and sherwood oil, the alcoholic acid volume(tric)fraction is 80%-100%, and the ratio of the volume of ethanol-water solution and sherwood oil is 3:1.
In the mixed solvent of above-mentioned ethanol-water solution and sherwood oil, the alcoholic acid volume(tric)fraction is 84%.
Step 2) the silica gel column chromatography partition method in is:
The developping agent that is made into sherwood oil and chloroform carries out thin-layer chromatography to filtered liq to be separated, and separates obtaining and Vitamin A Palmitate 1.7 M.I.U/Gram; Wherein the configuration proportion of developping agent PetroChina Company Limited. ether and chloroform is (1-5) by volume: 1.
The configuration proportion of above-mentioned developping agent PetroChina Company Limited.'s ether and chloroform is 2:1 by volume.
The present invention adopts the liquid-liquid extraction partition method to separate Vitamin A Palmitate 1.7 M.I.U/Gram and.Extraction is a kind of with liquid extraction agent immiscible with it two-pack of processing or polycomponent solution, thereby realizes the isolating mass transfer sepn process of component.Its ultimate principle is to utilize the difference of compound solubleness or partition ratio in the solvent of two kinds immiscible (or slightly solubles), and compound is transferred in the another kind of solvent from a kind of solvent.Through the repeated multiple times extraction, can title product and other components be separated as much as possible like this.It is very important that selection of Extractant seems in extraction, and it is big as far as possible in the partition ratio difference of extraction solvent two in mutually that suitable extraction agent is exactly each component of requirement, promptly selects coefficient big.
In addition, the present invention also adopts the silica gel column chromatography partition method to separate Vitamin A Palmitate 1.7 M.I.U/Gram and.Wherein the selection of eluting solvent is isolating key factor.By analyze can know treat that Vitamin A Palmitate 1.7 M.I.U/Gram is arranged in the isolating sample, the palmitinic acid of and trace.Wherein contain 1-COOH in the palmitinic acid molecular structure, polarity is stronger, and Vitamin A Palmitate 1.7 M.I.U/Gram polarity a little less than, especially with Vitamin A Palmitate 1.7 M.I.U/Gram polarity a little less than.Therefore, the organic solvent of selecting polarity to suit can be realized the separation and purification of Vitamin A Palmitate 1.7 M.I.U/Gram as wash-out moving phase.Select sherwood oil, benzene, chloroform, four kinds of organic solvents of ETHYLE ACETATE as developping agent according to the solvent polarity size, reaction solution is carried out the thin-layer chromatography separation test.The result shows: use any organic solvent all can not three kinds of materials be separated separately.Concern according to Vitamin A Palmitate 1.7 M.I.U/Gram and, palmitinic acid polar difference, select sherwood oil and two kinds of organic solvents of chloroform to join mutually.
The present invention selects two kinds of diverse ways to separate Vitamin A Palmitate 1.7 M.I.U/Gram, and the one, liquid-liquid separation has advantage simple to operate, low for equipment requirements; Another is that silica gel column chromatography separates, and utilizes this separation operating method can obtain the very high Vitamin A Palmitate 1.7 M.I.U/Gram of purity.
Description of drawings
Below in conjunction with accompanying drawing and embodiment the present invention is made further detailed description, but accompanying drawing and embodiment to should not be construed as be the qualification that the present invention is carried out.
Fig. 1 is the removal effect figure of palmitinic acid under the differing temps in the embodiment of the invention 1.
Fig. 2 is the impact effect figure of ethanol solubility to extracting in the embodiment of the invention 2.
Fig. 3 be in the embodiment of the invention 3 extraction times to the presentation graphs that influences of separating effect.
Embodiment
And palmitinic acid are following through the concrete reaction process of fixed lipase catalyzed synthesise vitamins A cetylate:
In the system of reaction back Vitamin A Palmitate 1.7 M.I.U/Gram, unreacted, palmitinic acid and solvent sherwood oil are arranged.
A) removal of palmitinic acid in the system
Earlier reaction solution is placed 20 h at 4 ℃ of refrigerators, remove by filter the palmitinic acid of partly precipitated.Supernatant after then will filtering is placed on-20 ℃ of refrigerator 8 h, refilters and removes sedimentary palmitinic acid.
B) Vitamin A Palmitate 1.7 M.I.U/Gram separation and purification
Residual and the polarity of micro-palmitinic acid there are differences in Vitamin A Palmitate 1.7 M.I.U/Gram and the system.So this research sets about carrying out the separation and purification of Vitamin A Palmitate 1.7 M.I.U/Gram from two aspects: (1) liquid-liquid extraction separates; (2) silica gel column chromatography separates.
C) liquid-liquid extraction separates
Select several kinds of discrepant solvents of polarity to join mutually, the reaction solution vacuum concentration of having removed palmitinic acid that takes a morsel adds the extraction solvent for preparing to evaporate to dryness, stirs several minutes with glass rod, then standing demix.Get phase (extraction phase) and following (extracting phase) mutually, performance liquid chromatography check and analysis respectively.Also to investigate the influence of extraction times in addition, select suitable extraction times, both meet the requirements of purity, guarantee to have the higher recovery again separating effect.
D) silica gel column chromatography separates
(1) selects suitable wash-out moving phase
With the reaction solution of having removed palmitinic acid with the kapillary point on the silica-gel plate of having dried; According to polarity size select several kinds of organic solvents (sherwood oil, benzene, chloroform and ETHYLE ACETATE) and wherein two kinds of solvents after joining mutually by a certain percentage carry out thin-layer chromatography as developping agent.According to the separating effect of several kinds of materials, select suitable developping agent promptly as the wash-out moving phase of silica gel column chromatography;
(2) silicagel column separates
1. claim glue 200-300 order silica gel, take by weighing the silica gel of 30-70 times of applied sample amount;
2. the sherwood oil that stirs the adding certain volume is stirred well to even in dried silica gel with glass rod;
3. adorn post and use the cotton jam-pack at the bottom of with post, add the sherwood oil of about 1/3 volume, load onto and hold the liquid ball, open the post lower piston, an impouring of homogenate is held in the liquid ball, let its natural subsidence, will stick in post jamb silica gel with sherwood oil and pour in the post;
4. after the compacting sedimentation is accomplished, in holding the liquid ball, add a certain amount of sherwood oil, connect the air pump pressurization then, the compacting of post bed;
5. go up appearance and adopt appearance on the dry method.Spissated sample is added in a certain amount of dried silica gel, fully stir, the normal temperature held lets and treats that isolating sample fully is adsorbed onto on the silica gel particle.Pour the silica gel that has adsorbed sample in the pillar into, in pillar, add one little cotton then above the layer of silica gel, guarantee that wash-out moving phase can not wash out the post bed;
6. cross post and in holding the liquid ball, add wash-out moving phase, the beginning wash-out with collecting.Per 3 ml collection, 1 pipe detects with thin-layer chromatography earlier, identical sample merged, and concentrated, lyophilize, performance liquid chromatography detects, and calculates purity.
E) Davitin A check and analysis
(1) the HPLC detection method of Davitin A
Get reaction solution 0.2 ml, be settled to 10 ml, draw 100 μ l sample introductions with the methyl alcohol dilution.Chromatographic column is the reverse post of waters Nova-Park C18 (3.9 * 150 mm, 4 μ m), and moving phase is 100% chromatogram methyl alcohol; 25 ℃ of column temperatures, detector are Agilent G1328B DAD detector, detect wavelength 327 nm; Flow velocity 1 ml/min, times 17.2 min loses shape;
(2) drafting of typical curve
Accurately take by weighing 0.082 g, be dissolved in the 10 ml sherwood oils.Dilute 100 times, 160 times, 640 times, 1000 times, 1280 times respectively with methyl alcohol.Performance liquid chromatography detects, and is X-coordinate with the units, and peak area is an ordinate zou drawing standard curve.Typical curve: y=50.691x+180.14 R2=0.9914;
(3) drafting of vitamin A palmitinic acid typical curve
Accurately take by weighing 0.068 g Vitamin A Palmitate 1.7 M.I.U/Gram, be dissolved in the 10 ml sherwood oils.Dilute 20 times, 40 times, 80 times, 160 times, 320 times respectively with methyl alcohol.Performance liquid chromatography detects, and is X-coordinate with the Vitamin A Palmitate 1.7 M.I.U/Gram units, and peak area is an ordinate zou drawing standard curve.Vitamin A Palmitate 1.7 M.I.U/Gram typical curve: y=41.115x-316.51 R2=0.9999;
(4) data processing
If the volumetric molar concentration that adds before the reaction is A (mol/L), it is B (mol/L) that reaction finishes to generate the Vitamin A Palmitate 1.7 M.I.U/Gram volumetric molar concentration in the system of back, and residual volumetric molar concentration is C (mol/L).
Descriptive statistics: value listed in the figure is MV, adopts standard deviation to represent the dispersion degree of observed value to MV among the figure.
Transformation efficiency: transformation efficiency=B/A * 100%; Degradation rate: degradation rate=1-B/A * 100%-C/A * 100%.
The enzyme reaction initial velocity: enzyme reaction initial velocity (g/ml/min)=reaction finishes Vitamin A Palmitate 1.7 M.I.U/Gram mass concentration/15 that the back generates.
Pillar reaction flow velocity: reaction flow velocity (ml/ unit's column volume/unit time)=reaction solution volume/column volume/reaction times.
Embodiment 1: the removal of palmitinic acid
And palmitinic acid are following through the concrete reaction process of fixed lipase catalyzed synthesise vitamins A cetylate:
In the system of reaction back Vitamin A Palmitate 1.7 M.I.U/Gram, unreacted, palmitinic acid and solvent sherwood oil are arranged.Sherwood oil and acetate boiling point are lower, in the spissated process of vacuum-evaporation, can form the steam volatilization, are prone to remove.The existence of excessive palmitinic acid can have influence on separating of Vitamin A Palmitate 1.7 M.I.U/Gram and, thereby will remove palmitinic acid earlier.Under the palmitinic acid low temperature in sherwood oil solubleness lower, can mass crystallization come out.Therefore, can adopt the method for freezing to remove palmitinic acid.The present invention specifically investigates the removal effect of cetylate respectively under 4 ℃ ,-20 ℃, the time of freezing is 4-20 hour, and the result is as shown in Figure 1.
As can be seen from Figure 1, under 4 ℃, along with the prolongation of time, the removal amount of palmitinic acid increases gradually; Under-20 ℃, along with the prolongation of time, the removal amount of palmitinic acid increased before this, then tended towards stability.The removal amount of-20 ℃ of following palmitinic acids is higher than the removal amount under 4 ℃.But under-20 ℃ of conditions; A large amount of palmitinic acids are separated out simultaneously; The crystal that forms can comprise partial solvent; The minimizing of solvent can cause separating out of Vitamin A Palmitate 1.7 M.I.U/Gram again, and the part Vitamin A Palmitate 1.7 M.I.U/Gram that the new palmitinic acid crystallization meeting that forms will be separated out wraps up, and then causes the loss of product Vitamin A Palmitate 1.7 M.I.U/Gram.Take all factors into consideration, select optimized strategy be with reaction solution at 4 ℃ of condition held 20 h, palmitinic acid can partly be separated out, filter paper filtering is removed the palmitinic acid crystallization.The reaction solution that to remove the part palmitinic acid again is at-20 ℃ of condition held 8 h, and filter paper filtering is removed the palmitinic acid crystallization again.Cause the loss of Vitamin A Palmitate 1.7 M.I.U/Gram in the time of can avoiding the palmitinic acid mass crystallization like this.
Embodiment 2: extracting and separating Vitamin A Palmitate 1.7 M.I.U/Gram and
Filtered liq to obtain among the embodiment 1 is a raw material, and the present invention is on the basis of the multiple organic solvent of test, and the mixed solvent of selecting ethanol-water solution and sherwood oil is as extraction solvent, and the result will not list at this.Concentration of ethanol is that volume parts has material impact to extraction, ethanol solubility is too high or too low all can cause in the extraction not stratified.Table 1 and Fig. 2 are the influence of ethanol volume parts to extraction:
Table 1 alcohol concn is to the influence of extraction
| Ethanol solubility | Experimental phenomena | The partition ratio | The Vitamin A Palmitate 1.7 M.I.U/Gram partition ratio | Select coefficient |
| 0% | Not stratified | - | - | - |
| 20% | Not stratified | - | - | - |
| 40% | Layering | 260.19 | 93.28 | 0.36 |
| 60% | Layering | 197.18 | 132.26 | 0.67 |
| 80% | Layering | 15.25 | 76.33 | 5 |
| 100% | Not stratified | - | - | - |
Can find out from table 1,, select coefficient to increase gradually along with the increase of ethanol volume parts.When the ethanol volume(tric)fraction is 80%, select coefficient to reach maximum; And when ethanol solubility is 100%, extract not stratified.Therefore be necessary the ethanol volume(tric)fraction is done thinner research between 80%-100%.
Found out by Fig. 2, select coefficient to increase along with the increase of ethanol volume(tric)fraction earlier, then descend, the ethanol volume(tric)fraction is just not stratified above 90%.When its volume(tric)fraction is 84%, select coefficient to reach maximum.Possible cause is in this system, and an amount of water can promote biphase layering in the extraction process, also can impel the phase transition like the polar phase separately of Vitamin A Palmitate 1.7 M.I.U/Gram and Vitamin A Palmitate 1.7 M.I.U/Gram.Therefore, selecting the ethanol volume(tric)fraction is 84% to be used for further research.
The proportioning of ethanol-water solution and sherwood oil also has very significant effects to effect of extracting, and suitable proportioning can make Vitamin A Palmitate 1.7 M.I.U/Gram and separate as far as possible.Table 2 is ethanol-water solution and the influence of sherwood oil proportioning to extracting:
Table 2 ethanol-water solution and sherwood oil proportioning are to the influence of extraction
| Ethanol-water solution and sherwood oil proportioning | Experimental phenomena | The partition ratio | The Vitamin A Palmitate 1.7 M.I.U/Gram partition ratio | Select coefficient |
| 5:1 | Not stratified | - | - | - |
| 4:1 | Layering | 7.36 | 70.56 | 9.59 |
| 3:1 | Layering | 8.75 | 148.23 | 16.94 |
| 2:1 | Not stratified | - | - | - |
| 1:1 | Not stratified | - | - | - |
| 1:2 | Not stratified | - | - | - |
| 1:3 | Not stratified | - | - | - |
Find out from table 2, when ethanol-water solution and sherwood oil proportioning are 3:1, Vitamin A Palmitate 1.7 M.I.U/Gram two mutually in partition ratio and partition ratio differ maximum, promptly select coefficient big.It is very important to extracting and separating and Vitamin A Palmitate 1.7 M.I.U/Gram that this also further specifies suitable water-content.Therefore, selecting ethanol-water solution and sherwood oil proportioning is 3:1.
The influence of extraction temperature
This experimental selection under three temperature such as normal temperature, 4 ℃ ,-20 ℃, extract experimental result such as table 3:
Table 3 temperature is to the influence of extraction
| Extraction temperature/℃ | The partition ratio | The Vitamin A Palmitate 1.7 M.I.U/Gram partition ratio | Vitamin A Palmitate 1.7 M.I.U/Gram concentration (g/ml) | Select coefficient |
| Normal temperature | 7.17 | 73.98 | 0.0106 | 10.32 |
| 4 | 7.76 | 88.82 | 0.0116 | 11.45 |
| -20 | 9.22 | 99.13 | 0.0093 | 10.77 |
It is generally acknowledged that temperature has material impact to extraction; The spread coefficient that the one side elevated temperature can increase solute helps extracting and separating; The rising of temperature may cause separated material inactivation or destroy on the other hand, especially some thermo-sensitivitys and thermolability material.The result of table 3 meets this basic law.When extraction temperature is 4 ℃, select coefficient maximum, promptly the extracting and separating effect is best, and the Vitamin A Palmitate 1.7 M.I.U/Gram degraded is minimum.Thereby select 4 ℃ of extraction temperature as subsequent technique.
Embodiment 3: the checking of extraction and separation technology
Through the experiment among the embodiment 2, tentatively confirmed the technology of extraction separation purification Vitamin A Palmitate 1.7 M.I.U/Gram: the ethanol volume(tric)fraction is 84%, ethanol-water solution: sherwood oil=3:1, and extraction temperature is 4 ℃.Under these processing condition, carry out the extracting and separating of continuous several times, experimental result is as shown in Figure 3.
Find out that from Fig. 3 along with the increase of continuous extraction number of times, the purity of Vitamin A Palmitate 1.7 M.I.U/Gram increases gradually, and the recovery reduces progressively.When continuous extraction 4 times, this moment, the purity of Vitamin A Palmitate 1.7 M.I.U/Gram was 98.3%, and the recovery is 65.2%.After this continuing increases extraction times again, though Vitamin A Palmitate 1.7 M.I.U/Gram purity slightly rises, the recovery reduces obviously.Comprehensive purity and the recovery two aspects consider that continuous extraction just can reach the requirement of food with Vitamin A Palmitate 1.7 M.I.U/Gram 4 times, and with this understanding, the recovery is also than higher.
Embodiment 4: silica gel column chromatography separates and Vitamin A Palmitate 1.7 M.I.U/Gram
Filtered liq to obtain among the embodiment 1 is a raw material, concerns according to Vitamin A Palmitate 1.7 M.I.U/Gram and, palmitinic acid polar difference, selects sherwood oil and two kinds of organic solvents of chloroform to join mutually, and experiment organic solvent proportioning is to isolating influence.
Be made into the developping agent of different ratios (1:1,2:1,3:1,4:1,5:1) with sherwood oil and chloroform, treat separating reaction liquid and carry out the thin-layer chromatography test, the proportioning of test sherwood oil and chloroform is to isolating influence.Can find out by experimental result, join mutually that along with the increase of sherwood oil content, Vitamin A Palmitate 1.7 M.I.U/Gram and, palmitinic acid separating effect are poor more with sherwood oil and chloroform.When sherwood oil: during chloroform=2:1, Vitamin A Palmitate 1.7 M.I.U/Gram Rf value is 0.81, Rf value 0.47; Palmitinic acid Rf value is 0.08; Three's Rf value differs greatly, and this explains that with this understanding Vitamin A Palmitate 1.7 M.I.U/Gram and other two kinds of separating substances are effective.So the solvent system of selection sherwood oil: chloroform=2:1 is used for silica gel column chromatography as wash-out moving phase.
Davitin A is prone to oxidized destruction in illumination and air, thereby in the separation and purification operation, will consider lucifuge and anti-oxidation.And separate the back condensing crystal also is the important ring in the whole process of production; According to general crystallization strategy; Add a spot of Vitamin A Palmitate 1.7 M.I.U/Gram powder in the Vitamin A Palmitate 1.7 M.I.U/Gram solvent system to be crystallized containing; To promote the crystallization of Vitamin A Palmitate 1.7 M.I.U/Gram, obtain the very high Vitamin A Palmitate 1.7 M.I.U/Gram finished product of purity.
The variation that is appreciated that a lot of details is possible, but therefore this do not run counter to scope of the present invention and spirit, and the those of ordinary skill of any affiliated technical field all should be regarded as not breaking away from the category of patent of the present invention to its suitable variation of doing.
Claims (6)
1. the separation purification method of Vitamin A Palmitate 1.7 M.I.U/Gram is characterized in that comprising the steps:
1) removal of palmitinic acid: earlier with and palmitinic acid through the reaction product of fixed lipase catalyzed synthesise vitamins A cetylate at 4 ℃ of condition held 20 h, separate out the palmitinic acid crystallization, remove by filter the palmitinic acid crystallization; Supernatant liquid after then will filtering is placed on 8 h under-20 ℃ of conditions, separates out the palmitinic acid crystallization again, removes by filter the palmitinic acid crystallization once more, obtains filtered liq;
2) separate Vitamin A Palmitate 1.7 M.I.U/Gram and: the filtered liq that obtains separates Vitamin A Palmitate 1.7 M.I.U/Gram and with liquid-liquid extraction partition method or silica gel column chromatography partition method.
2. the separation purification method of Vitamin A Palmitate 1.7 M.I.U/Gram according to claim 1 is characterized in that step 2) in the liquid-liquid extraction partition method be:
With the filtered liq that obtains with the mixed solvent of ethanol-water solution and sherwood oil as the extraction solvent extracting and separating, extraction temperature is 4 ℃.
3. the separation purification method of Vitamin A Palmitate 1.7 M.I.U/Gram according to claim 2 is characterized in that in the mixed solvent of said ethanol-water solution and sherwood oil, and the alcoholic acid volume(tric)fraction is 80%-100%, and the ratio of the volume of ethanol-water solution and sherwood oil is 3:1.
4. the separation purification method of Vitamin A Palmitate 1.7 M.I.U/Gram according to claim 3 is characterized in that in the mixed solvent of said ethanol-water solution and sherwood oil, and the alcoholic acid volume(tric)fraction is 84%.
5. the separation purification method of Vitamin A Palmitate 1.7 M.I.U/Gram according to claim 1 is characterized in that step 2) in the silica gel column chromatography partition method be:
The developping agent that is made into sherwood oil and chloroform carries out thin-layer chromatography to filtered liq to be separated, and separates obtaining and Vitamin A Palmitate 1.7 M.I.U/Gram; Wherein the configuration proportion of developping agent PetroChina Company Limited. ether and chloroform is (1-5) by volume: 1.
6. the separation purification method of Vitamin A Palmitate 1.7 M.I.U/Gram according to claim 5 is characterized in that the configuration proportion of said developping agent PetroChina Company Limited.'s ether and chloroform is 2:1 by volume.
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Cited By (5)
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| CN103191134A (en) * | 2013-04-26 | 2013-07-10 | 四川省惠达药业有限公司 | Pharmaceutical composition of lipid-soluble vitamin and preparation method thereof |
| CN103194511A (en) * | 2013-03-06 | 2013-07-10 | 清华大学 | Method of lipase-catalyzed synthesis of fatty acid ester of clindamycin |
| KR20160141310A (en) * | 2015-05-29 | 2016-12-08 | 주식회사 마크로케어 | Two-phase reaction system for production of long chain retinyl ester and Preparation Method of ong chain retinyl ester |
| CN112724059A (en) * | 2021-01-15 | 2021-04-30 | 万华化学集团股份有限公司 | Preparation method of vitamin A palmitate |
| CN114907402A (en) * | 2022-05-07 | 2022-08-16 | 万华化学(四川)有限公司 | Synthesis of organic phosphonic fatty acid type ionic liquid and application of organic phosphonic fatty acid type ionic liquid in separation process of vitamin A palmitate |
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| THIERRY MAUGARD等: "Study of Vitamin Ester Synthesis by Lipase-Catalyzed Transesterification in Organic Media", 《BIOTECHNOL.PROG》 * |
| 刘祥洪等: "一步法合成维生素A棕榈酸酯", 《广东化工》 * |
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| CN103194511A (en) * | 2013-03-06 | 2013-07-10 | 清华大学 | Method of lipase-catalyzed synthesis of fatty acid ester of clindamycin |
| CN103194511B (en) * | 2013-03-06 | 2014-12-24 | 清华大学 | Method of lipase-catalyzed synthesis of fatty acid ester of clindamycin |
| CN103191134A (en) * | 2013-04-26 | 2013-07-10 | 四川省惠达药业有限公司 | Pharmaceutical composition of lipid-soluble vitamin and preparation method thereof |
| CN103191134B (en) * | 2013-04-26 | 2014-02-12 | 四川省惠达药业有限公司 | Pharmaceutical composition of lipid-soluble vitamin and preparation method thereof |
| KR20160141310A (en) * | 2015-05-29 | 2016-12-08 | 주식회사 마크로케어 | Two-phase reaction system for production of long chain retinyl ester and Preparation Method of ong chain retinyl ester |
| KR101700909B1 (en) | 2015-05-29 | 2017-02-14 | 주식회사 마크로케어 | Two-phase reaction system for production of long chain retinyl ester and Preparation Method of ong chain retinyl ester |
| CN112724059A (en) * | 2021-01-15 | 2021-04-30 | 万华化学集团股份有限公司 | Preparation method of vitamin A palmitate |
| CN112724059B (en) * | 2021-01-15 | 2023-05-30 | 万华化学集团股份有限公司 | Preparation method of vitamin A palmitate |
| CN114907402A (en) * | 2022-05-07 | 2022-08-16 | 万华化学(四川)有限公司 | Synthesis of organic phosphonic fatty acid type ionic liquid and application of organic phosphonic fatty acid type ionic liquid in separation process of vitamin A palmitate |
| CN114907402B (en) * | 2022-05-07 | 2024-02-27 | 万华化学(四川)有限公司 | Synthesis of organic phosphine fatty acid type ionic liquid and application of organic phosphine fatty acid type ionic liquid in vitamin A palmitate separation process |
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