CN104447889A - Preparation method of high-purity D-ribose - Google Patents
Preparation method of high-purity D-ribose Download PDFInfo
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Abstract
The invention discloses a preparation method of high-purity D-ribose. The preparation method of the high-purity D-ribose comprises the following steps: (1) ultrafiltering, namely ultrafiltering to remove macromolecular impurities; (2) precipitating in an alcohol, namely concentrating an ultrafiltrate at reduced pressure to obtain the concentrated ultrafiltrate of the D-ribose, and adding methanol or ethanol to precipitate in the methanol or ethanol; (3) exchanging ions, namely feeding a supernatant into a cationic-anionic-cationic resin column, and energizing on the side of an ion exchange solution; (4) carrying out chromatographic separation, namely concentrating the ion exchange solution which is collected in the step (3) at reduced pressure, feeding the concentrated ion exchange solution into a chromatographic separation resin, and collecting a chromatographic separation solution; (5) concentrating and crystallizing, namely concentrating and crystallizing to obtain wet high-purity D-ribose; (6) drying stepwise, namely putting the wet high-purity D-ribose which is obtained in the step (5) into a drying oven, and drying stepwise to obtain the finished product (high-purity D-ribose); and (7) measuring the finished product, namely preparing a solution from the high-purity D-ribose which is obtained in the step (6), and measuring the solution by adopting the high performance liquid chromatography. The preparation method of the high-purity D-ribose has the advantage of good D-ribose separation effect, and the purity of the separated D-ribose is more than 99.9%.
Description
Technical field
The present invention relates to food and pharmaceutical chemistry technical field, be specifically related to a kind of preparation method of high purity D-ribose.
Background technology
D-ribose is a kind of five-carbon ring aldehydo sugar that nature exists, and being present in all life cell, is the important component part of nucleic acid, coenzyme and cytogene, is widely used in the industrial sectors such as medicine, food, feed.In pharmaceutical industry, D-ribose can in a large number for the synthesis of riboflavin, also can be used as the raw materials for production of analog, terpenoid, modified amino acid, rennin supressor etc. of the precursor of nucleoside medicine, steroid, prostaglandin(PG), VD.In recent years, D-ribose is widely applied again the synthesis at many antiviral and antitumor drugs newly, as the medicine etc. of formycin, pyrazofurin, F-methylinosine phosphoric acid, halogenation chrondroitin and some treatment ovarian cancers.
The preparation method of current D-ribose mainly contains chemical synthesis and biological fermentation process two kinds.Compared with chemical synthesis, biological fermentation process has the advantages such as yield is high, cost is low, good product quality, environmental pollution are little, is thus generally adopted in the industrial production of D-ribose.But biological fermentation process is produced D-ribose itself and also be there are some shortcomings, impurity in such as D-ribose fermented liquid is more, containing plurality of impurities such as mycelium, protein, pigment, five-carbon sugar, hexose regulating YIN and YANG ion and phosphoric acid ester, cause the separation of D-ribose, purification comparatively difficulty, be thus difficult to prepare highly purified D-ribose.Current D-ribose manufacturing enterprise generally adopts following technique to carry out removal of impurities and purifies: fermentation liquor flocculation or foreign material such as macromole such as membrane filtration removing thalline, protein etc., cross carbon post or add activated carbon decolorizing, then by anion-cation exchange resin removing different kinds of ions impurity, again through concentrating under reduced pressure, add the ethanol of certain volume, crystallization under certain condition obtains D-ribose product.Make the D-ribose produced in this way generally can reach the purity of 97% ~ 99%, but be difficult to the high purity reaching more than 99.8%, the requirement of medical material can not be met.
Summary of the invention
For problems of the prior art, the invention provides a kind of preparation method of high purity D-ribose, the purity of D-ribose can be improved, and greatly reduce production cost.
For achieving the above object, the present invention is by the following technical solutions:
A preparation method for high purity D-ribose, comprises the following steps:
(1) ultrafiltration: D-ribose fermented liquid uses molecular weight cut-off to be 10,000 ~ 50, the ultra-filtration membrane ultrafiltration of 000, impurity such as macromole such as removing mycelium and protein etc.;
(2) alcohol precipitation: the pH regulating D-ribose ultrafiltrated is 2 ~ 6, D-ribose ultrafiltration and concentration liquid is obtained through concentrating under reduced pressure, then methyl alcohol or ethanol is added toward D-ribose ultrafiltration and concentration liquid, successively through stirring, standing, alcohol precipitation, pour out supernatant liquor, so operate 2 ~ 4 times, collect supernatant liquor, D-ribose all can be dissolved in methanol solution, a large amount of impurity in D-ribose fermented liquid are then for insolubles resides in solid residue;
(3) from friendship: sun-male-female resin column sent into by supernatant liquor step (2) collected, then conductance is surveyed what collect from friendship liquid; Be specially, supernatant liquor step (2) collected first crosses positive resin, that collects crosses negative resin from friendship liquid is direct, from negative resin collect from friendship liquid after positive resin, that is collected the 3rd positive resin post surveys conductance, if conductance is at 30 below μ s/cm from friendship liquid, it is qualified to be then considered as, if conductance is at 30 more than μ s/cm, then defective from friendship liquid, need again to cross resin from friendship.
(4) chromatographic separation: step (3) is collected from friendship liquid concentrating under reduced pressure, then the liquid after concentrating under reduced pressure is sent into chromatographic separation resin, collects chromatographic separation liquid; Three phases before, during and after the chromatographic separation liquid collected is divided into, in earlier stage waste liquid and impure more part discard, mid-term D-ribose purity lower than 99% chromatographic separation liquid return again chromatographic separation resin continue be separated, later stage D-ribose purity higher than 99% chromatogram liquid collect.
(5) condensing crystal: by concentrated for the chromatographic separation liquid collected, crystallization, obtain high purity D-ribose wet product;
(6) gradient is dry: step (5) is obtained high purity D-ribose wet product and puts into loft drier, and gradient is dry, namely obtains high purity D-ribose finished product after oven dry;
(7) finished product measures: the high purity D-ribose finished product wiring solution-forming obtained by step (6), uses high-performance liquid chromatogram determination.
In described step (2), the concentration of D-ribose ultrafiltration and concentration liquid is 40% ~ 75%, and during alcohol precipitation, methyl alcohol or ethanol add with the volume of 1.0 times, 0.5 times, 0.25 times D-ribose ultrafiltration and concentration liquid respectively.
The vitriol oil is adopted to regulate the pH value of D-ribose ultrafiltrated in described step (2).
From handing over the conductance of liquid need control at 30 below μ s/cm in described step (3), ensure the purity from handing over liquid.
20% ~ 70% is evaporated to from friendship liquid in described step (4).
In described step (5), chromatographic separation liquid is concentrated into 20% ~ 50%, concentrated solution stirs cooling at 40 DEG C ~ 75 DEG C, according to the speed of 0.5 DEG C ~ 5 DEG C/h, concentrated solution is cooled to 10 DEG C ~ 35 DEG C, add 150 order ~ 300 order D-ribose crystal seeds, after a large amount of precipitation to be crystallized, be cooled to-10 DEG C ~ 10 DEG C by the speed of 3 DEG C ~ 5 DEG C/h again, then centrifugation, obtain high purity D-ribose wet product.
In described step (6), drying temperature is 35 DEG C ~ 60 DEG C, and gradient increased temperature is dried.
In described step (7), high purity D-ribose finished product is made into the solution that concentration is 1.0%.
Compared with prior art, advantage of the present invention is:
1. in the present invention, each step of D-ribose separating-purifying all uses alcohols to come as medium, instead of traditional water-ol is the mode of medium.Under traditional way, be generally that after fermented liquid ultrafiltration, direct mistake is from friendship, the qualified of collection carries out chromatographic separation from friendship liquid, or without chromatographic separation direct concentrating under reduced pressure, extremely substantially anhydrous state, then add dissolve with ethanol, in ethanol crystallization.By seeing above, traditional method only uses ethanol in last crystallisation step, the present invention all uses alcohols as medium, when carrying out each step, D-ribose solution does not need to change back and forth between water and alcohol, make not only can improve product purity in this way, also can save concentration and evaporation cost simultaneously; 2. comprehensive membrane ultrafiltration of the present invention, alcohol precipitation, from friendship and chromatographic technique, had more alcohol precipitation and chromatrographic separation step than traditional method, made the purity being separated rear D-ribose reach more than 99.9%, far above existing method for purifying and separating, met the requirement of medical material; 3. the alcohol precipitation step that the present invention is exclusive can remove a large amount of impurity, greatly reduce the processing pressure of ion isolation resin, cross from compared with friendship with the ultrafiltrated without alcohol precipitation, precipitation solution is crossed from handing over the ion exchange resin amount only needing 1/50, and only this step just can reduce the production cost of D-ribose greatly.4. the present invention has created a kind of novel D-ribose chromatographic separation mode, and that uses alcohol sugar soln to carry out chromatographic separation exactly, and compared with molasses sugar solution chromatographic separation, the separating effect of this mode is better, and the D-ribose chromatogram liquid purity obtained is higher.
Accompanying drawing explanation
Fig. 1 is the liquid chromatogram measuring result of the embodiment of the present invention 1.
Fig. 2 is the liquid chromatogram measuring result of the embodiment of the present invention 2.
Fig. 3 is the liquid chromatogram measuring result of the embodiment of the present invention 3.
Fig. 4 is the liquid chromatogram measuring result of the embodiment of the present invention 4.
Fig. 5 is the liquid chromatogram measuring result of the embodiment of the present invention 5.
Fig. 6 is the liquid chromatogram measuring result of the embodiment of the present invention 6.
Embodiment
Below by specific embodiment, the invention will be further described, but embodiment does not limit in any form the present invention, can not limit protection scope of the present invention with this.
embodiment 1
The preparation method of the present embodiment high purity D-ribose, comprises the following steps:
(1) ultrafiltration: by D-ribose content be 9.00% fermented liquid use molecular weight cut-off be 10,000 ultra-filtration membrane ultrafiltration, the removing macromole such as mycelium and protein impurity, (2) alcohol precipitation: measure the D-ribose ultrafiltrated that 2300ml content is 8.85%, add the 5.8g vitriol oil, pH is regulated to be 2.0, through being evaporated to 40%, obtained D-ribose ultrafiltration and concentration liquid, then methyl alcohol is added toward D-ribose ultrafiltration and concentration liquid, methyl alcohol divides three times and adds, respectively with 1.0 times, 0.5 times, the volume of 0.25 times of D-ribose ultrafiltration and concentration liquid adds, successively through stirring, leave standstill alcohol precipitation, pour out supernatant liquor, operation like this 3 times, D-ribose all can be dissolved in methanol solution, a large amount of impurity in D-ribose fermented liquid are then for insolubles resides in solid residue, collect supernatant liquor, (3) from friendship: sun-male-female resin column sent into by supernatant liquor step (2) collected, and precipitation solution has entered rear use methyl alcohol liftout, then conductance is surveyed what collect from friendship liquid, be specially, supernatant liquor step (2) collected first crosses positive resin, that collects crosses negative resin from friendship liquid is direct, from negative resin collect from friendship liquid after positive resin, that is collected the 3rd positive resin post surveys conductance from friendship liquid, what the 3rd resin column was collected surveys a conductance from the every 300ml of friendship liquid, measurement result is at 10.0 μ s/cm ~ 25.0 μ s/cm, and mixing conductance is 16.7 μ s/cm, collects complete from friendship liquid, amount volume be 2960ml from friendship liquid, survey content be 6.82%.(4) chromatographic separation: step (3) is collected from the concentration handing over liquid to be evaporated to 20%, then the liquid after concentrating under reduced pressure is sent into chromatographic separation resin, feed liquid has entered rear use methyl alcohol liftout, collects chromatographic separation liquid; Three phases before, during and after the chromatographic separation liquid collected is divided into, in earlier stage waste liquid and impure more part discard, mid-term D-ribose purity lower than 99% chromatographic separation liquid return again chromatographic separation resin continue be separated, later stage D-ribose purity higher than 99% chromatogram liquid collect; (5) condensing crystal: by concentrated for the chromatographic separation liquid collected, crystallization, the later stage chromatogram collected is collected liquid and is concentrated into the concentration that D-ribose content is 20%, concentrated solution stirs cooling at the temperature of 40 DEG C, 10 DEG C are cooled to according to the speed of 5 DEG C/h, add 150 order D-ribose crystal seeds, wait a large amount of precipitation to be crystallized, then be cooled to 5 DEG C by the speed of 3 DEG C/h, then centrifugation, obtains high purity D-ribose wet product; (6) gradient is dry: step (5) is obtained high purity D-ribose wet product and puts into loft drier, and gradient is dry, namely under the temperature condition of 35 DEG C, is warming up to 50 DEG C of oven dry gradually, namely obtains high purity D-ribose finished product after oven dry; (7) finished product measures: accurately take the high purity D-ribose finished product that 1.0000g step (6) is obtained, be made into 100ml solution, and use high-performance liquid chromatogram determination, as shown in Figure 1, the purity of gained D-ribose finished product is 99.99% to result.
Embodiment 2
The preparation method of the present embodiment high purity D-ribose, comprises the following steps:
(1) ultrafiltration: by D-ribose content be 10.01% fermented liquid use molecular weight cut-off be 30,000 ultra-filtration membrane ultrafiltration, the removing macromole such as mycelium and protein impurity, (2) alcohol precipitation: measure the D-ribose ultrafiltrated that 2000ml content is 9.82%, add the 2.5g vitriol oil, regulate pH to be 4.0, be evaporated to the concentration of 55%, divide the methyl alcohol adding 1.0 times, 0.5 times, 0.25 times volumes for three times respectively in proportion, successively through stirring, leave standstill alcohol precipitation, pour out supernatant liquor, so operation 3 times, D-ribose all can be dissolved in methanol solution, a large amount of impurity in D-ribose ultrafiltrated are then for insolubles resides in solid residue, (3) from friendship: precipitation solution directly crosses sun-male-female ion exchange resin column, precipitation solution has entered rear use methyl alcohol liftout, then conductance is surveyed what collect from friendship liquid, be specially, ion exchange resin column has three, precipitation solution first crosses first positive resin, that collects directly enters second negative resin from friendship liquid, that collects from negative resin enters the 3rd positive resin from friendship liquid, what the 3rd resin column was collected surveys a conductance from the every 300ml of friendship liquid, measurement result is at 13.2 μ s/cm ~ 18.5 μ s/cm, mixing conductance is 15.6 μ s/cm, collect complete from friendship liquid, amount volume is 3000ml, surveying content is 6.52%, (4) chromatographic separation: that collects is qualified in the concentration of friendship liquid through being evaporated to 45%, concentrated solution enters chromatographic separation resin, feed liquid has entered rear use methyl alcohol liftout, three phases before, during and after the chromatogram liquid collected is divided into, in earlier stage waste liquid and impure more part discard, mid-term D-ribose purity lower than 99% chromatogram liquid return again chromatographic separation resin continue be separated, later stage D-ribose purity higher than 99% chromatogram liquid collect, (5) condensing crystal: the later stage chromatogram liquid collected is concentrated into the concentration that D-ribose content is 35%, concentrated solution stirs cooling at the temperature of 55 DEG C, 25 DEG C are cooled to according to the speed of 3 DEG C/h, add 200 order D-ribose crystal seeds, etc. a large amount of precipitation to be crystallized, be cooled to 0 DEG C by the speed of 4 DEG C/h again, centrifugation, obtain high purity D-ribose wet product, (6) gradient is dry: the D-ribose wet product after rejection filter, puts into loft drier, is warming up to 60 DEG C of oven dry under the temperature condition of 35 DEG C gradually, obtain high purity D-ribose crystal finished product, (7) finished product measures: the high purity D-ribose finished product 1.0000g accurately taking step (6) obtained is made into 100ml solution, and use high-performance liquid chromatogram determination, as shown in Figure 2, the purity of gained D-ribose finished product is 99.96% to result.
Embodiment 3
The preparation method of the present embodiment high purity D-ribose, comprises the following steps:
(1) ultrafiltration: by D-ribose content be 11.00% fermented liquid use molecular weight cut-off be 50,000 ultra-filtration membrane ultrafiltration, the removing macromole such as mycelium and protein impurity; (2) alcohol precipitation: measure the D-ribose ultrafiltrated that 2000ml content is 10.86%, add the 1.2g vitriol oil, regulate pH to be 6.0, be evaporated to the concentration of 75%, divide the ethanol adding 1.0 times, 0.5 times, 0.25 times volumes for three times respectively in proportion, successively through stirring, leave standstill alcohol precipitation, pour out supernatant liquor, 3 times like this, D-ribose all can be dissolved with ethanolic soln, a large amount of impurity in D-ribose ultrafiltrated are then for insolubles resides in solid residue; (3) from friendship: precipitation solution directly crosses sun-male-female ion exchange resin column, precipitation solution has entered rear use ethanol liftout, then survey conductance by what collect from friendship liquid, be specially, ion exchange resin column has three, precipitation solution first crosses first positive resin, that collects directly enters second negative resin from friendship liquid, and that collects from negative resin enters the 3rd positive resin from friendship liquid, and what the 3rd resin column was collected amasss as 3000ml from friendship liquid, mixing conductance is 13.2 μ s/cm, and surveying content is 7.18%; (4) chromatographic separation: will collect from the concentration of friendship liquid through being evaporated to 70%, concentrated solution enters chromatographic separation resin, feed liquid has entered rear use ethanol liftout, three phases before, during and after the chromatogram liquid collected is divided into, in earlier stage waste liquid and impure more part discard, mid-term D-ribose purity lower than 99% chromatogram liquid return again chromatographic separation resin continue be separated, later stage D-ribose purity higher than 99% chromatogram liquid collect; (5) condensing crystal: the later stage chromatogram liquid collected is concentrated into the concentration that D-ribose content is 50%, concentrated solution stirs cooling at the temperature of 75 DEG C, 35 DEG C are cooled to according to the speed of 4 DEG C/h, add 300 order D-ribose crystal seeds, etc. a large amount of precipitation to be crystallized, be cooled to 10 DEG C by the speed of 5 DEG C/h again, centrifugation, obtain high purity D-ribose wet product; (6) gradient is dry: the D-ribose wet product after rejection filter, puts into loft drier, is warming up to 55 DEG C of oven dry under the temperature condition of 35 DEG C gradually, obtain high purity D-ribose crystal finished product; (7) finished product measures: accurately take the high purity D-ribose crystal finished product 1.0000g that step (6) is obtained, be made into 100ml solution, and use high-performance liquid chromatogram determination, as shown in Figure 3, the purity of gained D-ribose finished product is 99.92% to result.
Embodiment 4
The preparation method of the present embodiment high purity D-ribose, comprises the following steps:
(1) ultrafiltration: by D-ribose content be 10.60% fermented liquid use molecular weight cut-off be 40,000 ultra-filtration membrane ultrafiltration, the removing macromole such as mycelium and protein impurity; (2) alcohol precipitation: measure the D-ribose ultrafiltrated that 2000ml content is 10.46%, add the 1.9g vitriol oil, regulate pH to be 5.0, be evaporated to the concentration of 75%, divide the ethanol adding 1.0 times, 0.5 times, 0.25 times volumes for three times respectively in proportion, successively through stirring, leave standstill alcohol precipitation, pour out supernatant liquor, 3 times like this, D-ribose all can be dissolved with ethanolic soln, a large amount of impurity in D-ribose ultrafiltrated are then for insolubles resides in solid residue; (3) from friendship: precipitation solution directly crosses sun-male-female ion exchange resin column, precipitation solution has entered rear use ethanol liftout, then survey conductance by what collect from friendship liquid, be specially, ion exchange resin column has three, precipitation solution first crosses first positive resin, that collects directly enters second negative resin from friendship liquid, and that collects from negative resin enters the 3rd positive resin from friendship liquid, and what the 3rd resin column was collected amasss as 3000ml from friendship liquid, mixing conductance is 14.6 μ s/cm, and surveying content is 6.83%; (4) chromatographic separation: will collect from the concentration of friendship liquid through being evaporated to 50%, concentrated solution enters chromatographic separation resin, feed liquid has entered rear use ethanol liftout, three phases before, during and after the chromatogram liquid collected is divided into, in earlier stage waste liquid and impure more part discard, mid-term D-ribose purity lower than 99% chromatogram liquid return again chromatographic separation resin continue be separated, later stage D-ribose purity higher than 99% chromatogram liquid collect; (5) condensing crystal: the later stage chromatogram liquid collected is concentrated into the concentration that D-ribose content is 30%, concentrated solution stirs cooling at the temperature of 45 DEG C, 20 DEG C are cooled to according to the speed of 2 DEG C/h, add 250 order D-ribose crystal seeds, etc. a large amount of precipitation to be crystallized, be cooled to-5 DEG C by the speed of 4 DEG C/h again, centrifugation, obtain high purity D-ribose wet product; (6) gradient is dry: the D-ribose wet product after rejection filter, puts into loft drier, is warming up to 55 DEG C of oven dry under the temperature condition of 35 DEG C gradually, obtain high purity D-ribose crystal finished product; (7) finished product measures: accurately take the high purity D-ribose crystal finished product 1.0000g that step (6) is obtained, be made into 100ml solution, and use high-performance liquid chromatogram determination, as shown in Figure 4, the purity of gained D-ribose finished product is 99.95% to result.
Embodiment 5
The preparation method of the present embodiment high purity D-ribose, comprises the following steps:
(1) ultrafiltration: by D-ribose content be 9.80% fermented liquid use molecular weight cut-off be 30,000 ultra-filtration membrane ultrafiltration, the removing macromole such as mycelium and protein impurity; (2) alcohol precipitation: measure the D-ribose ultrafiltrated that 2000ml content is 9.66%, add the 3.7g vitriol oil, regulate pH to be 3.0, be evaporated to the concentration of 60%, divide the ethanol adding 1.0 times, 0.5 times, 0.25 times volumes for three times respectively in proportion, successively through stirring, leave standstill alcohol precipitation, pour out supernatant liquor, 3 times like this, D-ribose all can be dissolved with ethanolic soln, a large amount of impurity in D-ribose ultrafiltrated are then for insolubles resides in solid residue; (3) from friendship: precipitation solution directly crosses sun-male-female ion exchange resin column, precipitation solution has entered rear use ethanol liftout, then survey conductance by what collect from friendship liquid, be specially, ion exchange resin column has three, precipitation solution first crosses first positive resin, that collects directly enters second negative resin from friendship liquid, and that collects from negative resin enters the 3rd positive resin from friendship liquid, and what the 3rd resin column was collected amasss as 3000ml from friendship liquid, mixing conductance is 18.3 μ s/cm, and surveying content is 6.37%; (4) chromatographic separation: will collect from the concentration of friendship liquid through being evaporated to 30%, concentrated solution enters chromatographic separation resin, feed liquid has entered rear use ethanol liftout, three phases before, during and after the chromatogram liquid collected is divided into, in earlier stage waste liquid and impure more part discard, mid-term D-ribose purity lower than 99% chromatogram liquid return again chromatographic separation resin continue be separated, later stage D-ribose purity higher than 99% chromatogram liquid collect; (5) condensing crystal: the later stage chromatogram liquid collected is concentrated into the concentration that D-ribose content is 40%, concentrated solution stirs cooling at the temperature of 60 DEG C, 15 DEG C are cooled to according to the speed of 5 DEG C/h, add 180 order D-ribose crystal seeds, etc. a large amount of precipitation to be crystallized, be cooled to-10 DEG C by the speed of 5 DEG C/h again, centrifugation, obtain high purity D-ribose wet product; (6) gradient is dry: the D-ribose wet product after rejection filter, puts into loft drier, is warming up to 55 DEG C of oven dry under the temperature condition of 35 DEG C gradually, obtain high purity D-ribose crystal finished product; (7) finished product measures: accurately take the high purity D-ribose crystal finished product 1.0000g that step (6) is obtained, be made into 100ml solution, and use high-performance liquid chromatogram determination, as shown in Figure 5, the purity of gained D-ribose finished product is 99.95% to result.
Embodiment 6
The preparation method of the present embodiment high purity D-ribose, comprises the following steps:
(1) ultrafiltration: by D-ribose content be 10.82% fermented liquid use molecular weight cut-off be 20,000 ultra-filtration membrane ultrafiltration, the removing macromole such as mycelium and protein impurity; (2) alcohol precipitation: measure the D-ribose ultrafiltrated that 2000ml content is 10.63%, add the 1.8g vitriol oil, regulate pH to be 5.0, be evaporated to the concentration of 65%, divide the ethanol adding 1.0 times, 0.5 times, 0.25 times volumes for three times respectively in proportion, successively through stirring, leave standstill alcohol precipitation, pour out supernatant liquor, 3 times like this, D-ribose all can be dissolved with ethanolic soln, a large amount of impurity in D-ribose ultrafiltrated are then for insolubles resides in solid residue; (3) from friendship: precipitation solution directly crosses sun-male-female ion exchange resin column, precipitation solution has entered rear use ethanol liftout, then survey conductance by what collect from friendship liquid, be specially, ion exchange resin column has three, precipitation solution first crosses first positive resin, that collects directly enters second negative resin from friendship liquid, and that collects from negative resin enters the 3rd positive resin from friendship liquid, and what the 3rd resin column was collected amasss as 3000ml from friendship liquid, mixing conductance is 16.2 μ s/cm, and surveying content is 6.94%; (4) chromatographic separation: will collect from the concentration of friendship liquid through being evaporated to 60%, concentrated solution enters chromatographic separation resin, feed liquid has entered rear use ethanol liftout, three phases before, during and after the chromatogram liquid collected is divided into, in earlier stage waste liquid and impure more part discard, mid-term D-ribose purity lower than 99% chromatogram liquid return again chromatographic separation resin continue be separated, later stage D-ribose purity higher than 99% chromatogram liquid collect; (5) condensing crystal: the later stage chromatogram liquid collected is concentrated into the concentration that D-ribose content is 45%, concentrated solution stirs cooling at the temperature of 40 DEG C, 30 DEG C are cooled to according to the speed of 5 DEG C/h, add 150 order D-ribose crystal seeds, etc. a large amount of precipitation to be crystallized, be cooled to 0 DEG C by the speed of 5 DEG C/h again, centrifugation, obtain high purity D-ribose wet product; (6) gradient is dry: the D-ribose wet product after rejection filter, puts into loft drier, is warming up to 55 DEG C of oven dry under the temperature condition of 35 DEG C gradually, obtain high purity D-ribose crystal finished product; (7) finished product measures: accurately take the high purity D-ribose crystal finished product 1.0000g that step (6) is obtained, be made into 100ml solution, and use high-performance liquid chromatogram determination, as shown in Figure 6, the purity of gained D-ribose finished product is 99.94% to result.
Claims (8)
1. a preparation method for high purity D-ribose, is characterized in that, comprises the following steps:
(1) ultrafiltration: D-ribose fermented liquid uses molecular weight cut-off to be 10,000 ~ 50, the ultra-filtration membrane ultrafiltration of 000, removing macromole impurity, obtains D-ribose ultrafiltrated;
(2) alcohol precipitation: the pH regulating D-ribose ultrafiltrated is 2 ~ 6, obtains D-ribose ultrafiltration and concentration liquid, then add methyl alcohol or ethanol toward D-ribose ultrafiltration and concentration liquid through concentrating under reduced pressure, successively through stirring, standing, alcohol precipitation, pour out supernatant liquor, so operate 2 ~ 4 times, collect supernatant liquor;
(3) from friendship: sun-male-female resin column sent into by supernatant liquor step (2) collected, then conductance is surveyed what collect from friendship liquid;
(4) chromatographic separation: step (3) is collected from friendship liquid concentrating under reduced pressure, then the liquid after concentrating under reduced pressure is sent into chromatographic separation resin, collects chromatographic separation liquid;
(5) condensing crystal: by concentrated for the chromatographic separation liquid collected, crystallization, obtain high purity D-ribose wet product;
(6) gradient is dry: step (5) is obtained high purity D-ribose wet product and puts into loft drier, and gradient is dry, namely obtains high purity D-ribose finished product after oven dry;
(7) finished product measures: the high purity D-ribose finished product wiring solution-forming obtained by step (6), uses high-performance liquid chromatogram determination.
2. the preparation method of high purity D-ribose according to claim 1, it is characterized in that: in described step (2), the mass concentration of D-ribose ultrafiltration and concentration liquid is 40% ~ 75%, and during alcohol precipitation, methyl alcohol or ethanol add with the volume of 1.0 times, 0.5 times, 0.25 times D-ribose ultrafiltration and concentration liquid respectively.
3. the preparation method of high purity D-ribose according to claim 1, is characterized in that: adopt the vitriol oil to regulate the pH value of D-ribose ultrafiltrated in step (2) described in described step.
4. the preparation method of high purity D-ribose according to claim 1, is characterized in that: from handing over the conductance of liquid to control at 30 below μ s/cm in described step (3).
5. the preparation method of high purity D-ribose according to claim 1, is characterized in that: being evaporated to mass concentration from friendship liquid in described step (4) is 20% ~ 70%.
6. the preparation method of high purity D-ribose according to claim 1, it is characterized in that: in described step (5), chromatographic separation liquid is concentrated into mass concentration is 20 ~ 50%, concentrated solution stirs cooling at 30 DEG C ~ 75 DEG C, according to the speed of 0.5 DEG C ~ 5 DEG C/h, concentrated solution is cooled to 10 DEG C ~ 25 DEG C, add 150 order ~ 300 order D-ribose crystal seeds, after a large amount of precipitation to be crystallized, then according to the speed of 3 DEG C ~ 5 DEG C/h to-10 DEG C ~ 10 DEG C, then centrifugation, obtains high purity D-ribose wet product.
7. the preparation method of high purity D-ribose according to claim 1, is characterized in that: in described step (6), drying temperature is 35 DEG C ~ 60 DEG C, and gradient increased temperature is dried.
8. the preparation method of high purity D-ribose according to claim 1, is characterized in that: in described step (7), high purity D-ribose finished product is made into the solution that mass concentration is 1.0%.
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| CN201410786353.9A Pending CN104447889A (en) | 2014-12-18 | 2014-12-18 | Preparation method of high-purity D-ribose |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4904587A (en) * | 1987-09-10 | 1990-02-27 | Takeda Chemical Industries, Ltd. | Production of D-ribose |
| CN1234404A (en) * | 1999-03-22 | 1999-11-10 | 华东理工大学 | Method for separating D-ribose from fermentation liquor |
| CN1508138A (en) * | 2002-12-18 | 2004-06-30 | 诚志生命科技有限公司 | Method for extracting D-ribose crystal from fermented broth |
| CN1616473A (en) * | 2004-09-21 | 2005-05-18 | 山东省食品发酵工业研究设计院 | Method for separating and extracting D-ribose from fermented liquid by film separating technology |
| CN102241706A (en) * | 2010-12-31 | 2011-11-16 | 三达膜科技（厦门）有限公司 | D-ribose purification and separation method |
| CN103145771A (en) * | 2013-03-15 | 2013-06-12 | 江西诚志生物工程有限公司 | Method for extracting D-ribose from fermentation liquor by ultrafiltration and ion exchange technologies |
-
2014
- 2014-12-18 CN CN201410786353.9A patent/CN104447889A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4904587A (en) * | 1987-09-10 | 1990-02-27 | Takeda Chemical Industries, Ltd. | Production of D-ribose |
| CN1234404A (en) * | 1999-03-22 | 1999-11-10 | 华东理工大学 | Method for separating D-ribose from fermentation liquor |
| CN1508138A (en) * | 2002-12-18 | 2004-06-30 | 诚志生命科技有限公司 | Method for extracting D-ribose crystal from fermented broth |
| CN1616473A (en) * | 2004-09-21 | 2005-05-18 | 山东省食品发酵工业研究设计院 | Method for separating and extracting D-ribose from fermented liquid by film separating technology |
| CN102241706A (en) * | 2010-12-31 | 2011-11-16 | 三达膜科技（厦门）有限公司 | D-ribose purification and separation method |
| CN103145771A (en) * | 2013-03-15 | 2013-06-12 | 江西诚志生物工程有限公司 | Method for extracting D-ribose from fermentation liquor by ultrafiltration and ion exchange technologies |
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