CN114149477A - Crystallization method of high-purity vitamin B12 crystal and product thereof - Google Patents
Crystallization method of high-purity vitamin B12 crystal and product thereof Download PDFInfo
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- FDJOLVPMNUYSCM-WZHZPDAFSA-L cobalt(3+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(1r,2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2 Chemical compound [Co+3].N#[C-].N([C@@H]([C@]1(C)[N-]\C([C@H]([C@@]1(CC(N)=O)C)CCC(N)=O)=C(\C)/C1=N/C([C@H]([C@@]1(CC(N)=O)C)CCC(N)=O)=C\C1=N\C([C@H](C1(C)C)CCC(N)=O)=C/1C)[C@@H]2CC(N)=O)=C\1[C@]2(C)CCC(=O)NC[C@@H](C)OP([O-])(=O)O[C@H]1[C@@H](O)[C@@H](N2C3=CC(C)=C(C)C=C3N=C2)O[C@@H]1CO FDJOLVPMNUYSCM-WZHZPDAFSA-L 0.000 title claims abstract description 86
- 229930003779 Vitamin B12 Natural products 0.000 title claims abstract description 85
- 239000011715 vitamin B12 Substances 0.000 title claims abstract description 85
- 235000019163 vitamin B12 Nutrition 0.000 title claims abstract description 85
- 239000013078 crystal Substances 0.000 title claims abstract description 68
- 238000002425 crystallisation Methods 0.000 title claims abstract description 46
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- 230000008025 crystallization Effects 0.000 claims abstract description 37
- 239000007864 aqueous solution Substances 0.000 claims abstract description 28
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- 239000002245 particle Substances 0.000 claims abstract description 5
- 239000002994 raw material Substances 0.000 claims abstract description 3
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- 239000012528 membrane Substances 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000003480 eluent Substances 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 239000000706 filtrate Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000001728 nano-filtration Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
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- -1 polycyclic compound Chemical class 0.000 description 2
- OVARTBFNCCXQKS-UHFFFAOYSA-N propan-2-one;hydrate Chemical group O.CC(C)=O OVARTBFNCCXQKS-UHFFFAOYSA-N 0.000 description 2
- 238000005086 pumping Methods 0.000 description 2
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 2
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- 208000014644 Brain disease Diseases 0.000 description 1
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- 235000011114 ammonium hydroxide Nutrition 0.000 description 1
- 238000005571 anion exchange chromatography Methods 0.000 description 1
- 239000003957 anion exchange resin Substances 0.000 description 1
- 229940058548 b-12 resin Drugs 0.000 description 1
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- 125000002091 cationic group Chemical group 0.000 description 1
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- 229910017052 cobalt Inorganic materials 0.000 description 1
- 239000010941 cobalt Substances 0.000 description 1
- GUTLYIVDDKVIGB-UHFFFAOYSA-N cobalt atom Chemical compound [Co] GUTLYIVDDKVIGB-UHFFFAOYSA-N 0.000 description 1
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- WUPRCGRRQUZFAB-DEGKJRJSSA-N corrin Chemical group N1C2CC\C1=C\C(CC/1)=N\C\1=C/C(CC\1)=N/C/1=C\C1=NC2CC1 WUPRCGRRQUZFAB-DEGKJRJSSA-N 0.000 description 1
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- 235000013345 egg yolk Nutrition 0.000 description 1
- 210000002969 egg yolk Anatomy 0.000 description 1
- ZOOODBUHSVUZEM-UHFFFAOYSA-N ethoxymethanedithioic acid Chemical compound CCOC(S)=S ZOOODBUHSVUZEM-UHFFFAOYSA-N 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
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- 125000000168 pyrrolyl group Chemical group 0.000 description 1
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- 229920002477 rna polymer Polymers 0.000 description 1
- 235000010288 sodium nitrite Nutrition 0.000 description 1
- 235000010265 sodium sulphite Nutrition 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
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- 239000012991 xanthate Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H23/00—Compounds containing boron, silicon, or a metal, e.g. chelates, vitamin B12
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention discloses a crystallization method of high-purity vitamin B12 crystals, which comprises the following steps: step one, preparing a vitamin B12 acetone aqueous solution by taking a vitamin B12 crude product as a raw material, placing the solution in a crystallization container, and stirring the solution; step two, slowly and uniformly dripping an anti-solvent acetone into a crystallization container, and controlling the dripping flow rate to be 0.5-1.0 BV/h; after the dropwise addition is finished, quickly and uniformly dropwise adding an anti-solvent acetone into the crystallization container, and controlling the dropwise adding flow rate to be 3.0-3.5 BV/h; after the dropwise adding is finished, performing aging treatment; and step three, filtering and drying the product prepared in the step two. The vitamin B12 crystal prepared by the invention is in the shape of block particles and has the advantages of large size, small specific surface area, higher purity and better stability (difficult moisture absorption).
Description
Technical Field
The invention relates to the technical field of vitamin B12, in particular to a crystallization method of a high-purity vitamin B12 crystal and a product thereof.
Background
Vitamin B12, abbreviated as VB12, also known as cobalamin, having the molecular formula C63H88CoN14O14P, molecular weight 1355.38, is a polycyclic compound containing 3-valent cobalt, with 4 reduced pyrrole rings linked together to form 1 corrin ring, the only metal-element-containing vitamin.
Vitamin B12 is an important vitamin, is a coenzyme necessary in the synthesis process of ribonucleic acid and deoxyribonucleic acid, and is mainly present in animal food, such as liver, kidney, pig heart and the like, lean meat, fish, cow milk, egg yolk and the like. The lack of VB12 in vivo can cause pathological changes in the nervous system such as peripheral and central encephalopathies. In daily life, vitamin B12 can be used in the fields of health food and injection, and with the increasing requirements on food and medicine safety, preparing vitamin B12 with high purity and low impurity content becomes the object of the skilled in the art.
The crystallization process is one of the key means for purifying and removing the impurities of the vitamin B12, because the spreading agent of the vitamin B12 resin chromatography is acetone water solution, in addition, the vitamin B12 is not dissolved in acetone, and the acetone has the characteristic of being mutually soluble with water, the vitamin B12 mainly prepares a finished crystal product by the crystallization process of adding an anti-solvent acetone to change the solubility of feed liquid at present. The crystallization method has the advantages of high crystallization yield and easy recovery of a single organic solvent acetone, for example, Chinese patent application with application publication number CN111808158A discloses a preparation method of a vitamin B12 crude product, wherein the disclosed crystallization process is to add 8-10 times of anti-solvent acetone into a vitamin B12 filtrate. However, the vitamin B12 crystal prepared by the crystallization method is needle-shaped, small in size, easy to break in the crystallization process, large in specific surface area, easy to carry impurities, poor in crystal stability and easy to absorb moisture.
Disclosure of Invention
Aiming at the problems in the prior art, the invention discloses a method for crystallizing high-purity vitamin B12 crystal, which prepares blocky granular B12 crystal by adopting two-stage crystallization and accurately controlling the parameters of each stage of crystallization, and has the advantages of large size, small specific surface area, higher purity and better stability (difficult moisture absorption).
The specific technical scheme is as follows:
a crystallization method of high-purity vitamin B12 crystals, comprising:
step one, preparing a vitamin B12 acetone aqueous solution by taking a vitamin B12 crude product as a raw material, placing the solution in a crystallization container, and stirring the solution;
step two, slowly and uniformly dripping an anti-solvent acetone into a crystallization container, and controlling the dripping flow rate to be 0.5-1.0 BV/h; after the dropwise addition is finished, quickly and uniformly dropwise adding an anti-solvent acetone into the crystallization container, and controlling the dropwise adding flow rate to be 3.0-3.5 BV/h; after the dropwise adding is finished, performing aging treatment;
and step three, filtering and drying the product prepared in the step two to obtain the high-purity vitamin B12 crystal.
The crystallization method disclosed by the invention adopts a two-stage crystallization process and an aging process, and specifically, a slow uniform-speed dripping mode is adopted in the first stage, a fast uniform-speed dripping mode is adopted in the second stage, the dripping amount and the dripping flow rate of acetone in the two stages are respectively controlled, and finally, the aging treatment is carried out, so that the finally prepared vitamin B12 crystal is ensured to be in the form of blocky particles, and the finally prepared vitamin B12 crystal has the advantages of large size, small specific surface area, higher purity and better stability.
In the first step:
the vitamin B12 acetone aqueous solution is directly obtained by eluting the acetone aqueous solution when the vitamin B12 crude product is subjected to resin chromatography, or is prepared from the vitamin B12 crude product;
the vitamin B12 crude product is obtained by flocculating and hydrolyzing the fermentation liquor, filtering with a membrane, converting, removing impurities by ion exchange, carrying out macroporous resin chromatography, concentrating, crystallizing and drying. The preparation process comprises the following steps:
(1) flocculation hydrolysis of fermentation liquor: adding 1% equivalent of polyaluminium chloride and sodium sulfite into the fermentation liquor, and hydrolyzing at 90 ℃ for 30 min;
(2) membrane filtration and impurity removal: filtering the hydrolysate with ceramic membrane, and high pressure nano-filtering and concentrating the filtrate with 200Da nano-filtration membrane;
(3) and (3) positive columns: adjusting the pH of the nanofiltration concentrated solution to 3.0 by using 7% hydrochloric acid, then passing the nanofiltration concentrated solution through cobalamion cationic resin, eluting the resin by using 8.5% ammonia water, and immediately adjusting the pH of an eluent to 4-5 by using 7% hydrochloric acid;
(4) and (3) transformation: adding 2 equivalents of VB12 xanthate and sodium nitrite into the positive column eluent, and carrying out heat preservation and conversion for 2h at the temperature of 80 ℃;
(5) macroporous resin chromatography: microfiltration is carried out on the converted conversion solution through a ceramic membrane, then, filtrate passes through an XAD1180N macroporous resin chromatographic column, after washing and desalting by pure water, a 7-8% acetone water solution is used for spreading layers, VB12 components with high purity and concentration are collected, and the acetone solvent is removed through evaporation and concentration;
(6) anion exchange chromatography: after the concentrated VB12 solution passes through IRA900Cl anion exchange resin, immediately adjusting the pH value of the feed liquid to 5-7 by using a 4% sodium hydroxide solution;
(7) and (3) macroporous resin concentration: feeding the feed liquid after the negative column into an XAD1180N macroporous resin column, and eluting with 80% acetone aqueous solution to obtain the vitamin B12 acetone aqueous solution;
(8) and (3) crystallizing and drying: pouring the concentrated eluent into a three-mouth bottle, dropwise adding acetone with the volume 4 times that of the concentrated solution under stirring at room temperature, then putting the three-mouth bottle into a constant-temperature cooling tank, cooling to 0 ℃, and preserving heat for 1 h. And (3) carrying out suction filtration, collecting a filter cake, and drying in an oven at 50 ℃ for 3-6 h to obtain a VB12 crude product, wherein the purity of the VB12 crude product is 98.5%.
Preferably:
in the vitamin B12 acetone aqueous solution, the concentration of the vitamin B12 crude product is 10-50 g/L, and the concentration of acetone in the acetone aqueous solution is 50-80 vol% (volume ratio).
Preferably:
the temperature of the crystallization container is controlled to be 10-30 ℃, and the stirring speed is 50-300 rpm.
In the second step:
the dropping flow rate is 0.5-1.0 BV/h, namely the 1h flow rate is 0.5-1 time of the volume of the initial vitamin B12 acetone aqueous solution; the same explanation is also given for the later dripping flow rate of 3.0 to 3.5 BV/h.
Preferably:
when the anti-solvent acetone is slowly and uniformly dripped, the addition amount of the acetone is 2.0-3.5 times of the acetone aqueous solution of the vitamin B12 in the step one;
and (3) when the anti-solvent acetone is quickly and uniformly dripped, the addition amount of the acetone is 5.0-7.5 times of that of the vitamin B12 acetone aqueous solution in the step one.
In the second step, the flow rate and the dropping amount of the acetone during slow uniform dropping and the flow rate and the dropping amount during fast uniform dropping are controlled within the preferable ranges, so that the purity of the prepared vitamin B12 crystal can be ensured to reach more than 99%, and the crystallization yield is more than 93%.
Further preferably:
when the anti-solvent acetone is slowly and uniformly dripped, the addition amount of the acetone is 2.5-3.5 times of the acetone aqueous solution of the vitamin B12 in the step one;
and (3) when the anti-solvent acetone is quickly and uniformly dripped, the addition amount of the acetone is 6.5-7.5 times of that of the vitamin B12 acetone aqueous solution in the step one.
The crystallization yield of the product vitamin B12 crystal can be further improved to be higher than 96 percent by further optimizing the dropping amount of the acetone dropped twice.
Preferably, in the second step, the aging time is 1-3 h.
In the third step:
the filtration adopts a Buchner funnel;
and drying for 5-12 h at the temperature of 50-85 ℃ under a vacuum condition.
Under the above-mentioned continuously preferred process conditions, more preferably:
step two, slowly and uniformly dripping an anti-solvent acetone into a crystallization container, wherein the dripping flow rate is controlled to be 0.5BV/h, and the dripping amount is 2.5 times of the vitamin B12 acetone aqueous solution in the step one; after the dropwise addition, quickly and uniformly dropwise adding an anti-solvent acetone into the crystallization container, controlling the dropwise adding flow rate to be 3.0BV/h, wherein the dropwise adding amount is 6.5 times of the vitamin B12 acetone aqueous solution in the step one; and aging after the dropwise addition is finished.
The purity and the crystallization yield of the vitamin B12 crystal prepared by the more preferable process are both optimal.
The invention also discloses a high-purity vitamin B12 crystal prepared by the method, wherein the high-purity vitamin B12 crystal is blocky particles, the length-diameter ratio is 1-2, the length is 100-160 mu m, and the width is 60-120 mu m; the purity is more than 99%.
Compared with the prior art, the invention has the following beneficial effects:
the invention discloses a crystallization method of high-purity vitamin B12 crystals, which comprises the steps of adopting two-stage crystallization, accurately controlling the dropping amount and the dropping flow rate of acetone in each stage of crystallization, and finally carrying out aging treatment to prepare blocky granular B12 crystals, wherein the crystals have the advantages of large size, small specific surface area and better stability (difficult moisture absorption). More importantly, the preparation process of the invention can obtain the purity of more than 99 percent without sacrificing the production cost, such as increasing the high-resolution chromatography process to improve the purity.
Drawings
FIG. 1 is a microphotograph of vitamin B12 crystals prepared in example 1;
FIG. 2 is a microphotograph of vitamin B12 crystals prepared in example 2;
FIG. 3 is a microphotograph of vitamin B12 crystals prepared in comparative example 1;
fig. 4 is a microphotograph of vitamin B12 crystals prepared in comparative example 3.
Detailed Description
The present invention is further illustrated by the following examples, which are not intended to limit the scope of the invention.
Example 1
(1) Taking 200mL of vitamin B12 acetone aqueous solution obtained by chromatography, wherein the concentration of VB12 is 50000mg/L, the concentration of acetone is 50% (volume ratio), placing the solution in a 2L crystallization container, controlling the temperature of the crystallization container to be 30 ℃, and setting the stirring speed to be 50 rpm;
(2) measuring 500mL of acetone in a beaker, slowly dropwise adding the acetone into a crystallization container by using a peristaltic pump, and setting the dropwise adding speed of the peristaltic pump to be 0.5BV/h (100mL/h) until the dropwise adding is finished;
(3) weighing 1300mL of acetone in the beaker again, and accelerating the pumping speed of the peristaltic pump to 3.0BV/h (600mL/h) until the acetone is continuously aged for 1h after the acetone is dropwise added;
(4) filtering the crude vitamin B12 crystal product by using a Buchner funnel to obtain a wet vitamin B12 crystal product; and drying the VB12 wet product at 50 ℃ for 12 hours under vacuum condition to obtain a VB12 crystal finished product, and weighing.
The microphotograph of VB12 crystal prepared in this example is shown in FIG. 1, and when observing FIG. 1, it can be seen that the average length of the crystal was 120 μm, the average width was 65 μm, and the average aspect ratio was 1.85.
Example 2
(1) Dissolving 200mL of a VB12 crude product to obtain a vitamin B12 acetone aqueous solution, wherein the concentration of VB12 is 10000mg/L and the concentration of acetone is 80 percent (volume ratio), placing the solution in a 2L crystallization container, controlling the temperature of the crystallization container to be 10 ℃, and setting the stirring speed to be 300 rpm;
(2) weighing 700mL of acetone into a beaker, slowly dropwise adding the acetone into a crystallization container by using a peristaltic pump, and setting the dropwise adding speed of the peristaltic pump to be 1BV/h (200mL/h) until the dropwise adding is finished;
(3) weighing 1500mL of acetone into the beaker again, accelerating the pumping speed of the peristaltic pump to 3.5BV/h (700mL/h) until the acetone is dropwise added, and continuing to age for 3 h;
(4) filtering the crude vitamin B12 crystal product by using a Buchner funnel to obtain a wet vitamin B12 crystal product; and drying the VB12 wet product at 85 ℃ for 5 hours under vacuum condition to obtain a VB12 crystal finished product, and weighing.
The microphotograph of VB12 crystal prepared in this example is shown in FIG. 2, and when observing FIG. 2, it can be seen that the average length of the crystal was 155. mu.m, the average width was 117. mu.m, and the average aspect ratio was 1.32.
Example 3
The preparation process is basically the same as that of example 1, except that in step (2), the dropping speed of the peristaltic pump is adjusted to be 1.0 BV/h.
This example produced VB12 crystals having an average length of 116 μm, an average width of 105 μm and an average aspect ratio of 1.10.
Example 4
The preparation process is basically the same as that of example 1, except that 550mL of acetone is measured in step (2), and the dropping speed of a peristaltic pump is adjusted to be 0.8 BV/h.
The VB12 crystals prepared in this example had an average length of 112 μm, an average width of 97 μm and an average aspect ratio of 1.15.
Example 5
The preparation process is basically the same as that of example 1, except that 1300mL of acetone is measured in step (3), and the dropping speed of a peristaltic pump is adjusted to be 3.5 BV/h.
The VB12 crystals prepared in this example had an average length of 125 μm, an average width of 103 μm and an average aspect ratio of 1.21.
Example 6
The preparation process is basically the same as that of example 1, except that 1400mL of acetone is measured in step (3), and the dropping speed of a peristaltic pump is adjusted to be 3.2 BV/h.
The VB12 crystals prepared in this example had an average length of 124 μm, an average width of 86 μm and an average aspect ratio of 1.44.
Example 7
The preparation process is basically the same as that of example 1, except that in step (2), 400mL of acetone is measured, and the dropping speed of a peristaltic pump is adjusted to be 1.0 BV/h.
The crystals prepared in this example had an average length of 119 μm, an average width of 102 μm and an average aspect ratio of 1.17.
Example 8
The preparation process is basically the same as that of example 1, except that in step (3), 1000mL of acetone is measured, and the dropping speed of a peristaltic pump is adjusted to be 3.5 BV/h.
The crystals prepared in this comparative example had an average length of 122 μm, an average width of 88 μm, and an average aspect ratio of 1.39.
Comparative example 1
The preparation process is basically the same as that of example 1, except that in step (2), 600mL of acetone is measured, and the dropping speed of a peristaltic pump is adjusted to be 1.5 BV/h.
A photomicrograph of VB12 crystals prepared in this comparative example is shown in FIG. 3, and it can be seen from the photomicrograph that the morphology is also lumpy under the microscope, but the particles are small in size, the average length of the crystals is 51 μm, the average width is 45 μm, and the average aspect ratio is 1.13.
Comparative example 2
The preparation process is basically the same as that of example 1, except that in step (3), the dropping speed of the peristaltic pump is adjusted to 4.0 BV/h.
The crystals prepared in this comparative example had an average length of 83 μm, an average width of 79 μm, and an average aspect ratio of 1.05.
Comparative example 3 (conventional process):
200mL of the chromatographed vitamin B12 aqueous acetone solution of the same lot as in example 1, in which the concentration of VB12 was 50000mg/L and the concentration of acetone was 50% (by volume), was placed in a 2L crystallization vessel, the temperature of the crystallization vessel was controlled at 30 ℃ and the stirring rate was set at 50 rpm. Acetone was slowly added dropwise to the crystallization vessel using a peristaltic pump until turbidity appeared. Slowly cooling to 4 ℃, the cooling rate is 5 min/DEG C, continuously dripping 800mL of acetone at the dripping speed of 0.5BV/h, and after finishing dripping, preserving heat and aging at 2 ℃ for 2 h. After aging, filtering the vitamin B12 product by using a Buchner funnel to obtain a wet crystallized vitamin B12 product; and vacuum drying the VB12 wet product at 50 ℃ for 12h, and weighing the VB12 crystal finished product.
The microphotograph of VB12 crystals prepared in this comparative example is shown in FIG. 4, and it is understood that needle-shaped crystals were prepared by the conventional crystallization process, the crystals had an average length of 75 μm, an average width of 6 μm, and an average aspect ratio of 12.50.
And (3) performance testing:
(1) crystal color: the color of the crystals was visually checked and compared with the existing product (comparative example 3 product).
(2)
(3) Purity: referring to the method provided in the united states pharmacopeia 2017 edition, a VB12 aqueous solution with the concentration of 1mg/mL is prepared, after liquid phase separation, the peak area ratio of VB12 of a sample is calculated according to a peak area normalization method, and the liquid phase conditions are shown in the following table 1:
TABLE 1
The properties of the products prepared in the above examples and comparative examples, respectively, are listed in the following table 2.
TABLE 2
Remarking: the higher the crystal color score in the table, the darker the color, the better the quality of the VB12 crystals.
And (3) stability testing:
to further prove that the vitamin B12 crystal prepared by the present invention has excellent stability, the vitamin B12 crystal prepared in example 1 and comparative example 3 respectively is mixed with water to prepare 1mg/mL aqueous solution, and the acid resistance, alkali resistance, high temperature resistance and light resistance are respectively examined, and the test conditions and performance data are favorable in the following Table 3.
TABLE 3
Comparing the data in table 3, it can be determined that the vitamin B12 crystal prepared by the present invention has more excellent stability, which may be due to the blocky shape, large size, small specific surface area and higher purity.
Claims (10)
1. A method for crystallizing high-purity vitamin B12 crystals, comprising:
step one, preparing a vitamin B12 acetone aqueous solution by taking a vitamin B12 crude product as a raw material, placing the solution in a crystallization container, and stirring the solution;
step two, slowly and uniformly dripping an anti-solvent acetone into a crystallization container, and controlling the dripping flow rate to be 0.5-1.0 BV/h; after the dropwise addition is finished, quickly and uniformly dropwise adding an anti-solvent acetone into the crystallization container, and controlling the dropwise adding flow rate to be 3.0-3.5 BV/h; after the dropwise adding is finished, performing aging treatment;
and step three, filtering and drying the product prepared in the step two to obtain the high-purity vitamin B12 crystal.
2. The method for crystallizing high-purity vitamin B12 crystals according to claim 1, wherein in step one:
the vitamin B12 acetone aqueous solution is directly obtained by eluting the acetone aqueous solution when the vitamin B12 crude product is subjected to resin chromatography, or is prepared from the vitamin B12 crude product;
in the vitamin B12 acetone aqueous solution, the concentration of the vitamin B12 crude product is 10-50 g/L, and the concentration of acetone in the acetone aqueous solution is 50-80 vol%.
3. The method for crystallizing high-purity vitamin B12 crystals according to claim 1, wherein in the first step, the temperature of the crystallization vessel is controlled to 10 to 30 ℃ and the stirring speed is 50 to 300 rpm.
4. The method for crystallizing high-purity vitamin B12 crystals according to claim 1, wherein in step two:
when the anti-solvent acetone is slowly and uniformly dripped, the addition amount of the acetone is 2.0-3.5 times of the acetone aqueous solution of the vitamin B12 in the step one;
and (3) when the anti-solvent acetone is quickly and uniformly dripped, the addition amount of the acetone is 5.0-7.5 times of that of the vitamin B12 acetone aqueous solution in the step one.
5. The method for crystallizing high-purity vitamin B12 crystals according to claim 4, wherein in step two:
when the anti-solvent acetone is slowly and uniformly dripped, the addition amount of the acetone is 2.5-3.5 times of the acetone aqueous solution of the vitamin B12 in the step one;
and (3) when the anti-solvent acetone is quickly and uniformly dripped, the addition amount of the acetone is 6.5-7.5 times of that of the vitamin B12 acetone aqueous solution in the step one.
6. The method for crystallizing high-purity vitamin B12 crystals according to claim 1, wherein the aging time in step two is 1 to 3 hours.
7. The method for crystallizing high-purity vitamin B12 crystals according to claim 1, wherein in step three, the filtration is performed using a Buchner funnel.
8. The method for crystallizing high-purity vitamin B12 crystals according to claim 1, wherein the drying is performed at 50-85 ℃ under vacuum for 5-12 hours in step III.
9. The method for crystallizing high-purity vitamin B12 crystals according to claim 1, wherein the crystallization yield is more than 93%.
10. A high purity vitamin B12 crystal prepared according to the method of any one of claims 1-9, wherein:
the high-purity vitamin B12 crystal is a blocky particle, the average length-diameter ratio is 1-2, the average length is 100-160 mu m, and the average width is 60-120 mu m; the purity is more than 99%.
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