CN116621724A - Oxytetracycline extraction method for reducing content of impurity 2-acetyl-2-desamidooxytetracycline - Google Patents
Oxytetracycline extraction method for reducing content of impurity 2-acetyl-2-desamidooxytetracycline Download PDFInfo
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- CN116621724A CN116621724A CN202310896692.1A CN202310896692A CN116621724A CN 116621724 A CN116621724 A CN 116621724A CN 202310896692 A CN202310896692 A CN 202310896692A CN 116621724 A CN116621724 A CN 116621724A
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- oxytetracycline
- terramycin
- acetyl
- deamidated
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- 239000004100 Oxytetracycline Substances 0.000 title claims abstract description 65
- IWVCMVBTMGNXQD-PXOLEDIWSA-N oxytetracycline Chemical compound C1=CC=C2[C@](O)(C)[C@H]3[C@H](O)[C@H]4[C@H](N(C)C)C(O)=C(C(N)=O)C(=O)[C@@]4(O)C(O)=C3C(=O)C2=C1O IWVCMVBTMGNXQD-PXOLEDIWSA-N 0.000 title claims abstract description 65
- 229960000625 oxytetracycline Drugs 0.000 title claims abstract description 65
- 235000019366 oxytetracycline Nutrition 0.000 title claims abstract description 65
- IWVCMVBTMGNXQD-UHFFFAOYSA-N terramycin dehydrate Natural products C1=CC=C2C(O)(C)C3C(O)C4C(N(C)C)C(O)=C(C(N)=O)C(=O)C4(O)C(O)=C3C(=O)C2=C1O IWVCMVBTMGNXQD-UHFFFAOYSA-N 0.000 title claims abstract description 65
- 239000012535 impurity Substances 0.000 title claims abstract description 42
- 238000000605 extraction Methods 0.000 title claims abstract description 29
- KIPLYOUQVMMOHB-MXWBXKMOSA-L [Ca++].CN(C)[C@H]1[C@@H]2[C@@H](O)[C@H]3C(=C([O-])[C@]2(O)C(=O)C(C(N)=O)=C1O)C(=O)c1c(O)cccc1[C@@]3(C)O.CN(C)[C@H]1[C@@H]2[C@@H](O)[C@H]3C(=C([O-])[C@]2(O)C(=O)C(C(N)=O)=C1O)C(=O)c1c(O)cccc1[C@@]3(C)O Chemical compound [Ca++].CN(C)[C@H]1[C@@H]2[C@@H](O)[C@H]3C(=C([O-])[C@]2(O)C(=O)C(C(N)=O)=C1O)C(=O)c1c(O)cccc1[C@@]3(C)O.CN(C)[C@H]1[C@@H]2[C@@H](O)[C@H]3C(=C([O-])[C@]2(O)C(=O)C(C(N)=O)=C1O)C(=O)c1c(O)cccc1[C@@]3(C)O KIPLYOUQVMMOHB-MXWBXKMOSA-L 0.000 claims abstract description 103
- 229940063650 terramycin Drugs 0.000 claims abstract description 101
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims abstract description 23
- 238000000034 method Methods 0.000 claims abstract description 23
- 235000003891 ferrous sulphate Nutrition 0.000 claims abstract description 20
- 239000011790 ferrous sulphate Substances 0.000 claims abstract description 20
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims abstract description 20
- 239000000047 product Substances 0.000 claims description 72
- 238000000855 fermentation Methods 0.000 claims description 43
- 230000004151 fermentation Effects 0.000 claims description 43
- 238000003756 stirring Methods 0.000 claims description 41
- 239000007788 liquid Substances 0.000 claims description 33
- 238000005406 washing Methods 0.000 claims description 32
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims description 30
- 238000001914 filtration Methods 0.000 claims description 24
- 238000001035 drying Methods 0.000 claims description 23
- RZFBEFUNINJXRQ-UHFFFAOYSA-M sodium ethyl xanthate Chemical compound [Na+].CCOC([S-])=S RZFBEFUNINJXRQ-UHFFFAOYSA-M 0.000 claims description 21
- 230000003113 alkalizing effect Effects 0.000 claims description 18
- 239000003795 chemical substances by application Substances 0.000 claims description 18
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 18
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 18
- 229960001763 zinc sulfate Drugs 0.000 claims description 18
- 239000000706 filtrate Substances 0.000 claims description 17
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 claims description 16
- ZDYUUBIMAGBMPY-UHFFFAOYSA-N oxalic acid;hydrate Chemical compound O.OC(=O)C(O)=O ZDYUUBIMAGBMPY-UHFFFAOYSA-N 0.000 claims description 13
- 239000012530 fluid Substances 0.000 claims description 10
- 235000006408 oxalic acid Nutrition 0.000 claims description 10
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 8
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 8
- 239000013078 crystal Substances 0.000 claims description 8
- 238000007865 diluting Methods 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 8
- 235000010265 sodium sulphite Nutrition 0.000 claims description 8
- 239000011347 resin Substances 0.000 claims description 7
- 229920005989 resin Polymers 0.000 claims description 7
- 159000000000 sodium salts Chemical class 0.000 claims description 5
- ZOOODBUHSVUZEM-UHFFFAOYSA-N ethoxymethanedithioic acid Chemical compound CCOC(S)=S ZOOODBUHSVUZEM-UHFFFAOYSA-N 0.000 claims description 4
- 239000012991 xanthate Substances 0.000 claims description 4
- 230000001376 precipitating effect Effects 0.000 claims description 2
- CWYNVVGOOAEACU-UHFFFAOYSA-N Fe2+ Chemical compound [Fe+2] CWYNVVGOOAEACU-UHFFFAOYSA-N 0.000 abstract description 11
- 229910001448 ferrous ion Inorganic materials 0.000 abstract description 11
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 abstract description 9
- 229910001447 ferric ion Inorganic materials 0.000 abstract description 9
- 239000003638 chemical reducing agent Substances 0.000 abstract description 7
- 238000007254 oxidation reaction Methods 0.000 abstract description 7
- 230000020477 pH reduction Effects 0.000 abstract description 7
- 238000006728 Cope elimination reaction Methods 0.000 abstract description 4
- 230000003647 oxidation Effects 0.000 abstract description 4
- -1 phenol lactone Chemical class 0.000 abstract description 4
- 238000005303 weighing Methods 0.000 description 12
- 239000007791 liquid phase Substances 0.000 description 11
- 238000001514 detection method Methods 0.000 description 8
- 230000000052 comparative effect Effects 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 230000007423 decrease Effects 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 229910000358 iron sulfate Inorganic materials 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- NFVZIERLAZUYBQ-UHFFFAOYSA-N [K].[Zn] Chemical compound [K].[Zn] NFVZIERLAZUYBQ-UHFFFAOYSA-N 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000003379 elimination reaction Methods 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 239000000276 potassium ferrocyanide Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 235000010378 sodium ascorbate Nutrition 0.000 description 2
- PPASLZSBLFJQEF-RKJRWTFHSA-M sodium ascorbate Substances [Na+].OC[C@@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RKJRWTFHSA-M 0.000 description 2
- 229960005055 sodium ascorbate Drugs 0.000 description 2
- PPASLZSBLFJQEF-RXSVEWSESA-M sodium-L-ascorbate Chemical compound [Na+].OC[C@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RXSVEWSESA-M 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 241000270711 Malaclemys terrapin Species 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 230000001775 anti-pathogenic effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 125000002147 dimethylamino group Chemical group [H]C([H])([H])N(*)C([H])([H])[H] 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000010587 phase diagram Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000013225 prussian blue Substances 0.000 description 1
- 239000002516 radical scavenger Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 150000003512 tertiary amines Chemical class 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C231/00—Preparation of carboxylic acid amides
- C07C231/22—Separation; Purification; Stabilisation; Use of additives
- C07C231/24—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C2603/00—Systems containing at least three condensed rings
- C07C2603/02—Ortho- or ortho- and peri-condensed systems
- C07C2603/40—Ortho- or ortho- and peri-condensed systems containing four condensed rings
- C07C2603/42—Ortho- or ortho- and peri-condensed systems containing four condensed rings containing only six-membered rings
- C07C2603/44—Naphthacenes; Hydrogenated naphthacenes
- C07C2603/46—1,4,4a,5,5a,6,11,12a- Octahydronaphthacenes, e.g. tetracyclines
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The application provides an oxytetracycline extraction method for reducing the content of impurity 2-acetyl-2-deamidated oxytetracycline, which is characterized in that a strong reducing agent ferrous sulfate is added in an acidification stage in the initial stage of extraction, and the introduced ferrous ions and ferric ions can inhibit oxidation reaction after Cope elimination reaction and inhibit oxidation to form a phenol lactone, so that the generation of impurity 2-acetyl-2-deamidated oxytetracycline is reduced, the content of 2-acetyl-2-deamidated oxytetracycline is reduced, and the purity of oxytetracycline is improved; compared with the existing extraction method, the method can reduce the content of the impurity 2-acetyl-2-deamidated terramycin in the terramycin finished product, improve the purity of the terramycin finished product, avoid the influence of introducing new impurities on the subsequent operation and the product quality, and has high industrial application feasibility, and belongs to the technical field of terramycin extraction.
Description
Technical Field
The application relates to the technical field of oxytetracycline extraction, in particular to an oxytetracycline extraction method for reducing impurity 2-acetyl-2-deamidated oxytetracycline content.
Background
The terramycin is light yellow crystalline powder, is slightly soluble in ethanol, is slightly soluble in water, is stable in air, and is gradually dark in color when exposed to light. The oxytetracycline has broad-spectrum anti-pathogenic microorganism effect, and can be used as a rapid bacteriostat, and has bactericidal effect on certain bacteria at high concentration, so that the research on the oxytetracycline extraction method has great significance.
The existing terramycin extraction method mainly extracts terramycin from terramycin fermentation liquor, but byproducts and terramycin degradation products are generated in the process of extracting terramycin from terramycin fermentation liquor, hydrogen at the 5a position and hydroxyl at the 6 position of terramycin are subjected to trans elimination reaction to form hydrogen bonds in the initial acidification stage (pH < 2) of extraction, dehydrated terramycin is generated, the dehydrated terramycin is further degraded to form stereoisomerism alpha-aspartmycin and beta-aspartmycin, tertiary amine of alpha-aspartmycin and beta-aspartmycin is subjected to Ke Pu elimination reaction (Cope elimination reaction), dimethylamino is removed by oxidation, then the 7-formylating is performed to form a phenol lactone, and the phenol lactone is further reacted to form a deamide derivative 2-acetyl-2-deamide terramycin. The 2-acetyl-2-desamideomycin has poor antibacterial effect, and the content thereof is required to be less than 2.0% in pharmacopoeia, but since the structure of 2-acetyl-2-desamideomycin is very similar to that of oxytetracycline, it is difficult to be removed during purification, resulting in lower purity of oxytetracycline. This problem is to be solved.
Disclosure of Invention
In order to solve the technical problems, the technical scheme adopted by the application is to provide the oxytetracycline extraction method for reducing the content of impurity 2-acetyl-2-desamideomycin, so as to solve the technical problems of higher content of impurity 2-acetyl-2-desamideomycin and lower oxytetracycline purity in the existing oxytetracycline extraction method.
The embodiment of the application provides an oxytetracycline extraction method for reducing the content of impurity 2-acetyl-2-deamidated oxytetracycline, which comprises the following steps:
(1) Acidizing the terramycin fermentation broth by oxalic acid, stirring, then adding ferrous sulfate, and continuously stirring to obtain an acidized broth;
(2) Adding sodium xanthate and zinc sulfate into the acidizing fluid, stirring and standing for treatment to obtain a purified fluid;
(3) Diluting the purified solution with oxalic acid water, and then filtering and washing to obtain filtrate and washing liquid;
(4) Decolorizing the filtrate and the washing liquid to obtain decolorized liquid;
(5) Adding an alkalizing agent into the decolorized solution, stirring, precipitating crystals in the stirring process, filtering the materials after stirring to obtain a wet product, and drying the wet product to obtain the terramycin finished product.
Preferably, in step (1), the terramycin broth is acidified with oxalic acid to a pH <2 and stirred for 30min.
Preferably, in the step (1), the addition amount of the ferrous sulfate is 1.0-1.5%g/mL based on the volume of the terramycin fermentation broth, and stirring is continued for 30min.
Preferably, in the step (2), the added amount of the sodium ascorbate is 4.0-4.8 permillage g/mL based on the volume of the terramycin fermentation broth, and the added amount of the zinc sulfate is 2/3 of the weight of the sodium ascorbate.
Preferably, in the step (2), the stirring time is 30min, and the standing time is 1h.
Preferably, in the step (3), the pH value of the oxalic acid water is 2.5-3.5, the oxalic acid water is diluted to the titer of 10000-12000U/mL, the filtering mode is plate frame filtering, the washing mode is top washing, and the washing liquid in the washing process is oxalic acid water with the pH value of 2.5-3.5.
Preferably, in the step (5), the alkalizing agent is 15% ammonia water containing 2% sodium sulfite, the pH value of the decolorized solution is adjusted to 4.7-4.9 by adding the alkalizing agent, and the stirring time is 60min.
Preferably, in the step (5), the filtering mode is plate-frame filtering, the drying temperature is 50-100 ℃, and the quality of the terramycin finished product reaches constant weight.
The beneficial effects of the application are as follows: the application provides an oxytetracycline extraction method for reducing impurity 2-acetyl-2-deamidated oxytetracycline content, which comprises the steps of adding a strong reducing agent in an acidification stage in the initial stage of extractionFerrous sulfate, the ferrous ions and the ferric ions introduced can inhibit oxidation reaction after Cope elimination reaction and inhibit oxidation to form the terrapin, so that the generation of impurity 2-acetyl-2-deamidated terramycin is reduced, the content of 2-acetyl-2-deamidated terramycin is reduced, and the purity of terramycin is improved; in addition, in the purification stage, the introduced ferrous ions and ferric ions and the original ferric ions in the terramycin fermentation broth react with sodium salt of the yellow blood under an acidic environment to generate a precipitate: 4Fe 3+ +3Fe(CN) 6 4- →Fe 4 [Fe(CN) 6 ] 3 ∈so as to remove ferrous ions and ferric ions, and the balance of ferrous ions react with zinc sulfate to generate gel-like zinc potassium ferrocyanide double salt: 2K 4 Fe(CN) 6 +3ZnSO 4 →K 2 Zn 3 [Fe(CN) 6 ] 2 ↓+3K 2 SO 4 The proteins in the adsorbed terramycin fermentation liquor are precipitated together, so that secondary removal of ferrous ions is realized, and the influence of introduction of new impurities on subsequent operation and product quality due to addition of ferrous sulfate is avoided; compared with the existing extraction method, the method can reduce the content of the impurity 2-acetyl-2-deamidated terramycin in the terramycin finished product, improve the purity of the terramycin finished product, avoid the influence of introducing new impurities on the subsequent operation and the product quality, and has high industrial application feasibility.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are needed in the embodiments or the description of the prior art will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and that other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a process flow diagram of an oxytetracycline extraction method for reducing the impurity 2-acetyl-2-desamidooxytetracycline content of the present application;
FIG. 2 is a liquid-phase spectrum of the terramycin product prepared in example 3 of the application;
FIG. 3 is a liquid-phase diagram of the oxytetracycline product prepared in comparative example 1.
Detailed Description
In order to make the technical problems, technical schemes and beneficial effects to be solved more clear, the application is further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the application. The raw materials and the devices used in the application are conventional commercial products unless specified, and the methods used in the application are conventional methods unless specified.
Example 1
An oxytetracycline extraction method for reducing the content of impurity 2-acetyl-2-desamidooxytetracycline, comprising the following steps:
(1) Weighing 73L of terramycin fermentation liquor with the titer of 26622.40U/mL, acidizing the terramycin fermentation liquor by oxalic acid until the pH value is 1.96, stirring for 30min, adding 73.02g of ferrous sulfate (the addition amount of the ferrous sulfate is 1.0 per mill g/mL based on the volume of the terramycin fermentation liquor) into the terramycin fermentation liquor, and continuously stirring for 30min to obtain an acidized liquor;
(2) 292.09g of sodium xanthate (the adding amount of the sodium xanthate is 4.0 per mill g/mL based on the volume of the terramycin fermentation broth) and 194.73g of zinc sulfate (the adding amount of the zinc sulfate is 2/3 of the weight of the sodium xanthate) are added into the acidizing fluid, and the mixture is stirred for 30min and then is subjected to standing treatment for 1h, so as to obtain a purified liquid;
(3) Diluting the purified solution by oxalic acid water with the pH value of 3.1 to the titer of 10685.5U/ml, and then carrying out plate frame filtration and top washing to obtain filtrate and top washing liquid;
(4) Decolorizing the filtrate and the top washing liquid by using a 122# resin tank to obtain decolorized liquid;
(5) 15% ammonia water containing 2% sodium sulfite is used as an alkalizing agent, the alkalizing agent is added into the decolorized solution, the pH value of the decolorized solution is adjusted to 4.9, and the decolorized solution is stirred for 60min, and crystals are separated out in the stirring process; filtering the materials in a plate frame after stirring to obtain a wet product, then drying the wet product at 65 ℃, taking out and weighing the wet product for many times in the drying process until the mass of the wet product reaches constant weight to obtain 717.49g of finished terramycin, and carrying out liquid phase detection on the finished terramycin product to obtain the finished terramycin product with the purity of 97.13% and the impurity 2-acetyl-2-deamidated terramycin content of 0.87%; the process flow is shown in figure 1.
Example 2
An oxytetracycline extraction method for reducing the content of impurity 2-acetyl-2-desamidooxytetracycline, comprising the following steps:
(1) Weighing 73L of terramycin fermentation liquor with the titer of 24570.20U/mL, acidizing the terramycin fermentation liquor by oxalic acid until the pH value is 1.87, stirring for 30min, adding 87.63g of ferrous sulfate (the addition amount of the ferrous sulfate is 1.2 per mill g/mL based on the volume of the terramycin fermentation liquor) into the terramycin fermentation liquor, and continuously stirring for 30min to obtain the acidized liquor;
(2) 292.29g of sodium xanthate (the adding amount of the sodium xanthate is 4.0 per mill g/mL based on the volume of the terramycin fermentation broth) and 194.86g of zinc sulfate (the adding amount of the zinc sulfate is 2/3 of the weight of the sodium xanthate) are added into the acidizing fluid, and the mixture is stirred for 30min and then is subjected to standing treatment for 1h, so as to obtain a purified liquid;
(3) Diluting the purified solution by oxalic acid water with the pH value of 3.2 to the titer of 10775.8U/ml, and then carrying out plate frame filtration and top washing to obtain filtrate and top washing liquid;
(4) Decolorizing the filtrate and the top washing liquid by using a 122# resin tank to obtain decolorized liquid;
(5) 15% ammonia water containing 2% sodium sulfite is used as an alkalizing agent, the alkalizing agent is added into the decolorized solution, the pH value of the decolorized solution is adjusted to 4.9, and the decolorized solution is stirred for 60min, and crystals are separated out in the stirring process; and (3) carrying out plate-frame filtration on the materials after stirring to obtain a wet product, then drying the wet product at 65 ℃, taking out and weighing the wet product for multiple times in the drying process until the mass of the wet product reaches constant weight to obtain 695.02g of terramycin finished product, carrying out liquid phase detection on the terramycin finished product, and measuring that the purity of terramycin in the finished product is 97.45% and the content of impurity 2-acetyl-2-deamidated terramycin is 0.82%.
Example 3
An oxytetracycline extraction method for reducing the content of impurity 2-acetyl-2-desamidooxytetracycline, comprising the following steps:
(1) Weighing 73L of terramycin fermentation liquor with the titer of 25976.63U/mL, acidizing the terramycin fermentation liquor by oxalic acid until the pH value is 1.85, stirring for 30min, adding 109.51g of ferrous sulfate (the addition amount of the ferrous sulfate is 1.5 per mill g/mL based on the volume of the terramycin fermentation liquor) into the terramycin fermentation liquor, and continuously stirring for 30min to obtain the acidized liquor;
(2) 292.56g of sodium xanthate (the adding amount of the sodium xanthate is 4.0 per mill g/mL based on the volume of the terramycin fermentation broth) and 195.99g of zinc sulfate (the adding amount of the zinc sulfate is 2/3 of the weight of the sodium xanthate) are added into the acidizing fluid, and the mixture is stirred for 30min and then is subjected to standing treatment for 1h, so as to obtain a purified liquid;
(3) Diluting the purified solution by oxalic acid water with the pH value of 3.1 to the titer of 11369.67U/ml, and then carrying out plate frame filtration and top washing to obtain filtrate and top washing liquid;
(4) Decolorizing the filtrate and the top washing liquid by using a 122# resin tank to obtain decolorized liquid;
(5) 15% ammonia water containing 2% sodium sulfite is used as an alkalizing agent, the alkalizing agent is added into the decolorized solution, the pH value of the decolorized solution is adjusted to 4.85, and the decolorized solution is stirred for 60min, and crystals are separated out in the stirring process; and (3) carrying out plate-frame filtration on the materials after stirring to obtain a wet product, then drying the wet product at 65 ℃, taking out and weighing the wet product for multiple times in the drying process until the mass of the wet product reaches constant weight to obtain 700.24g of terramycin finished product, carrying out liquid phase detection on the terramycin finished product, and measuring that the purity of terramycin in the finished product is 97.57% and the content of impurity 2-acetyl-2-deamidated terramycin is 0.81%.
As is clear from examples 1 to 3, as the amount of the iron sulfate as a strong reducing agent added in the acidification stage increases from 1.0 to 1.2 g/mL, the content of 2-acetyl-2-desamidooxytetracycline as an impurity in the oxytetracycline product decreases, and as the amount of the iron sulfate as a strong reducing agent increases from 1.2 to 1.5 g/mL, the content of 2-acetyl-2-desamidooxytetracycline as an impurity in the oxytetracycline product still decreases, but the decrease is not large, and the purity of oxytetracycline still increases, and the increase is not large, so that, from an economical point of view, the amount of the added iron sulfate is preferably 1.2 g/mL based on the volume of the oxytetracycline fermentation broth.
Example 4
An oxytetracycline extraction method for reducing the content of impurity 2-acetyl-2-desamidooxytetracycline, comprising the following steps:
(1) Weighing 73L of terramycin fermentation liquor with the titer of 27325.68U/mL, acidizing the terramycin fermentation liquor by oxalic acid until the pH value is 1.77, stirring for 30min, adding 73.12g of ferrous sulfate (the addition amount of the ferrous sulfate is 1.0 per mill g/mL based on the volume of the terramycin fermentation liquor) into the terramycin fermentation liquor, and continuously stirring for 30min to obtain the acidized liquor;
(2) 321.24g of sodium xanthate (the adding amount of the sodium xanthate is 4.4 per mill g/mL based on the volume of the terramycin fermentation broth) and 213.16g of zinc sulfate (the adding amount of the zinc sulfate is 2/3 of the weight of the sodium xanthate) are added into the acidizing fluid, and the mixture is stirred for 30min and then is subjected to standing treatment for 1h, so as to obtain a purified liquid;
(3) Diluting the purified solution with oxalic acid water with pH of 2.7 to a titer of 11305.15U/ml, and then carrying out plate frame filtration and top washing to obtain filtrate and top washing liquid;
(4) Decolorizing the filtrate and the top washing liquid by using a 122# resin tank to obtain decolorized liquid;
(5) 15% ammonia water containing 2% sodium sulfite is used as an alkalizing agent, the alkalizing agent is added into the decolorized solution, the pH value of the decolorized solution is adjusted to 4.75, and the decolorized solution is stirred for 60min, and crystals are separated out in the stirring process; and (3) carrying out plate and frame filtration on the materials after stirring to obtain a wet product, then drying the wet product at 65 ℃, taking out and weighing the wet product for multiple times in the drying process until the mass of the wet product reaches constant weight to obtain 733.29g of terramycin finished product, carrying out liquid phase detection on the terramycin finished product, and measuring that the purity of terramycin in the finished product is 97.16% and the content of impurity 2-acetyl-2-deamidated terramycin is 0.85%.
Example 5
An oxytetracycline extraction method for reducing the content of impurity 2-acetyl-2-desamidooxytetracycline, comprising the following steps:
(1) Weighing 73L of terramycin fermentation liquor with the titer of 25873.16U/mL, acidizing the terramycin fermentation liquor by oxalic acid until the pH value is 1.75, stirring for 30min, adding 72.99g of ferrous sulfate (the addition amount of the ferrous sulfate is 1.0 per mill g/mL based on the volume of the terramycin fermentation liquor) into the terramycin fermentation liquor, and continuously stirring for 30min to obtain the acidized liquor;
(2) 350.41g of sodium xanthate (the adding amount of the sodium xanthate is 4.8 per mill g/mL based on the volume of the terramycin fermentation broth) and 235.66g of zinc sulfate (the adding amount of the zinc sulfate is 2/3 of the weight of the sodium xanthate) are added into the acidizing fluid, and the mixture is stirred for 30min and then is subjected to standing treatment for 1h, so as to obtain a purified liquid;
(3) Diluting the purified solution with oxalic acid water with pH of 2.7 to a titer of 10856.65U/ml, and then carrying out plate frame filtration and top washing to obtain filtrate and top washing liquid;
(4) Decolorizing the filtrate and the top washing liquid by using a 122# resin tank to obtain decolorized liquid;
(5) 15% ammonia water containing 2% sodium sulfite is used as an alkalizing agent, the alkalizing agent is added into the decolorized solution, the pH value of the decolorized solution is adjusted to 4.8, and the decolorized solution is stirred for 60min, and crystals are separated out in the stirring process; and (3) carrying out plate-frame filtration on the materials after stirring to obtain a wet product, then drying the wet product at 65 ℃, taking out and weighing the wet product for multiple times in the drying process until the mass of the wet product reaches constant weight to obtain 697.30g of terramycin finished product, carrying out liquid phase detection on the terramycin finished product, and measuring that the purity of terramycin in the finished product is 97.37% and the content of impurity 2-acetyl-2-deamidated terramycin is 0.83%.
As is clear from examples 1 and 4 to 5, as the amount of the scavenger sodium salt of xanthate added in the purification stage increases, the content of 2-acetyl-2-desamidooxytetracycline as an impurity in the oxytetracycline product gradually decreases, and the oxytetracycline purity gradually increases, so that the amount of sodium salt of xanthate added is preferably 4.8%g/mL based on the volume of oxytetracycline fermentation broth.
Comparative example 1
An oxytetracycline extraction method for reducing the content of impurity 2-acetyl-2-desamidooxytetracycline, comprising the following steps:
(1) Weighing 73L of terramycin fermentation liquor with the titer of 26454.60U/mL, acidizing the terramycin fermentation liquor by oxalic acid until the pH value is 1.86, and stirring for 30min to obtain an acidized liquor;
(2) 295.15g of sodium xanthate (the adding amount of the sodium xanthate is 4.0 per mill g/mL based on the volume of the terramycin fermentation broth) and 194.78g of zinc sulfate (the adding amount of the zinc sulfate is 2/3 of the weight of the sodium xanthate) are added into the acidizing fluid, and the mixture is stirred for 30min and then is subjected to standing treatment for 1h, so as to obtain a purified liquid;
(3) Diluting the purified solution with oxalic acid water with pH of 2.75 to a titer of 11886.8U/ml, and then carrying out plate frame filtration and top washing to obtain filtrate and top washing liquid;
(4) Decolorizing the filtrate and the top washing liquid by using a 122# resin tank to obtain decolorized liquid;
(5) 15% ammonia water containing 2% sodium sulfite is used as an alkalizing agent, the alkalizing agent is added into the decolorized solution, the pH value of the decolorized solution is adjusted to 4.75, and the decolorized solution is stirred for 60min, and crystals are separated out in the stirring process; and (3) carrying out plate-frame filtration on the materials after stirring to obtain a wet product, then drying the wet product at 65 ℃, taking out and weighing the wet product for multiple times in the drying process until the mass of the wet product reaches constant weight to obtain 713.92g of terramycin finished product, carrying out liquid phase detection on the terramycin finished product, and measuring that the purity of terramycin in the finished product is 95.95% and the content of impurity 2-acetyl-2-deamidated terramycin is 1.44%.
As can be seen from the examples 1 and the comparative example 1, the addition of the strong reducing agent ferrous sulfate in the acidification stage can obviously reduce the content of the impurity 2-acetyl-2-deamidated oxytetracycline in the oxytetracycline finished product and obviously improve the purity of oxytetracycline in the finished product.
Please refer to table 1, which is a summary of experimental data and experimental results of examples 1-5 and comparative example 1.
Table 1 summary of experimental data and experimental results for examples 1-5 and comparative example 1
As shown in Table 1, the addition of the strong reducing agent ferrous sulfate in the acidification stage can obviously reduce the content of the impurity 2-acetyl-2-deamidated terramycin in the terramycin finished product by more than 0.57%, obviously improve the purity of terramycin and improve the purity by more than 1.18%.
The terramycin finished product prepared in the example 3 is subjected to liquid phase detection, the liquid phase chromatogram is shown in figure 2, and as can be seen from figure 2, the terramycin content in the terramycin finished product is 97.57%, and the impurity 2-acetyl-2-deamidated terramycin content is 0.81%; the terramycin finished product prepared in the comparative example 1 is subjected to liquid phase detection, the liquid chromatogram is shown in figure 3, and as can be seen from figure 3, the terramycin content in the terramycin finished product is 95.95%, and the impurity 2-acetyl-2-deamidated terramycin content is 1.44%; as can be seen from FIGS. 2 and 3, the oxytetracycline product extracted by the extraction method of the present application has high purity, and the content of impurity 2-acetyl-2-desamidooxytetracycline can be significantly reduced.
In summary, the application provides an oxytetracycline extraction method for reducing the content of impurity 2-acetyl-2-deamidated oxytetracycline, wherein a strong reducing agent ferrous sulfate is added in an acidification stage in the initial stage of extraction, and the introduced ferrous ions and ferric ions can inhibit oxidation reaction after Cope elimination reaction and inhibit oxidation to form a phenol lactone, so that the generation of impurity 2-acetyl-2-deamidated oxytetracycline is reduced, the content of 2-acetyl-2-deamidated oxytetracycline is reduced, and the purity of oxytetracycline is improved; in addition, in the purification stage, the introduced ferrous ions, the ferric ions and the original ferric ions in the terramycin fermentation broth react with sodium xanthate under an acidic environment to generate precipitates, so that the removal of the ferrous ions and the ferric ions is realized, the rest ferrous ions react with zinc sulfate to generate gel-like zinc potassium ferrocyanide double salts, proteins in the terramycin fermentation broth are adsorbed to precipitate together, the secondary removal of the ferrous ions is realized, and the influence of the introduction of new impurities to subsequent operation and product quality due to the addition of ferrous sulfate is avoided; compared with the existing extraction method, the method can reduce the content of the impurity 2-acetyl-2-deamidated terramycin in the terramycin finished product, improve the purity of the terramycin finished product, avoid the influence of introducing new impurities on the subsequent operation and the product quality, has high industrial application feasibility, and can be widely applied to the technical field of terramycin extraction.
It should be noted that: when the wet product is dried, the drying temperature is selected to be 50-100 ℃, the drying temperature is lower, the drying time is longer, the drying temperature is higher, the drying time is shorter, and the drying temperature can be specifically adjusted according to actual conditions, for example, the drying temperature is set to be 65 ℃.
The above embodiments are only for illustrating the technical solution of the present application, and not for limiting the same; although the application has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present application, and are intended to be included in the scope of the present application.
Claims (9)
1. An oxytetracycline extraction method for reducing the content of impurity 2-acetyl-2-desamidooxytetracycline, comprising the following steps:
(1) Acidizing the terramycin fermentation broth by oxalic acid, stirring, then adding ferrous sulfate, and continuously stirring to obtain an acidized broth;
(2) Adding sodium xanthate and zinc sulfate into the acidizing fluid, stirring and standing for treatment to obtain a purified fluid;
(3) Diluting the purified solution with oxalic acid water, and then filtering and washing to obtain filtrate and washing liquid;
(4) Decolorizing the filtrate and the washing liquid to obtain decolorized liquid;
(5) Adding an alkalizing agent into the decolorized solution, stirring, precipitating crystals in the stirring process, filtering the materials after stirring to obtain a wet product, and drying the wet product to obtain the terramycin finished product.
2. The method for extracting oxytetracycline with reduced content of 2-acetyl-2-deamidated oxytetracycline as claimed in claim 1, wherein in step (1), the oxytetracycline fermentation broth is acidified with oxalic acid to pH <2 and stirred for 30min.
3. The method for extracting oxytetracycline with reduced content of 2-acetyl-2-deamidated oxytetracycline as defined in claim 1, wherein in step (1), the addition amount of ferrous sulfate is 1.0-1.5%g/mL based on the volume of oxytetracycline fermentation broth, and stirring is continued for 30min.
4. The method for extracting oxytetracycline with reduced content of 2-acetyl-2-deamidated oxytetracycline as defined in claim 1, wherein in the step (2), the added amount of sodium salt of xanthate is 4.0-4.8%o g/mL based on the volume of oxytetracycline fermentation broth, and the added amount of zinc sulfate is 2/3 of the weight of sodium salt of xanthate.
5. The method for extracting oxytetracycline with reduced content of 2-acetyl-2-deamidated oxytetracycline as described in claim 1, wherein in step (2), stirring time is 30min and standing time is 1h.
6. The method for extracting oxytetracycline with reduced content of 2-acetyl-2-deamidated oxytetracycline as defined in claim 1, wherein in the step (3), the pH value of oxalic acid water is 2.5-3.5, the titer of the purified solution is 10000-12000U/mL, the filtering mode is plate-frame filtering, the washing mode is top washing, and the washing liquid in the washing is oxalic acid water with the pH value of 2.5-3.5.
7. The method for extracting oxytetracycline with reduced content of 2-acetyl-2-deamidated oxytetracycline as described in claim 1, wherein in step (4), 122# resin tank is used to decolorize the filtrate and wash solution.
8. The method for extracting terramycin by reducing the content of 2-acetyl-2-deamidated terramycin as defined in claim 1, wherein in the step (5), the alkalizing agent is 15% ammonia water containing 2% sodium sulfite, the pH value of the decolorized solution is adjusted to 4.7-4.9 by adding the alkalizing agent, and the stirring time is 60min.
9. The method for extracting oxytetracycline with reduced content of 2-acetyl-2-deamidated oxytetracycline as defined in claim 1, wherein in step (5), the filtering mode is plate-frame filtering, the drying temperature is 50-100 ℃, and the oxytetracycline is dried until the quality of oxytetracycline finished product reaches constant weight.
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