Method for purifying acetylisovaleryltylosin tartrate

Abstract

The invention relates to a method for purifying acetylisovaleryltylosin tartrate, which comprises the following steps: adsorbing, desorbing and separating a crude acetylisovaleryltylosin tartrate dissolving solution by macroporous resin, decolorizing by anion resin, and finally carrying out nanofiltration concentration and spray drying to obtain a pure acetylisovaleryltylosin tartrate product. The method has the advantages of strong selectivity, high separation purity, substantial improvement of the purity of the acetylisovaleryltylosin tartrate product, good product yield, regeneration and reuse of the filter material, no use of flammable and explosive organic solvents, strong safety, cost saving and consumption reduction, and is suitable for large-scale production.

Description

Technical Field

[0001] The invention belongs to the technical field of biological fermentation and extraction, and particularly relates to a method for purifying tyvalosin tartrate. Background Art

[0002] Tylvalosin, also known as acetylisovaleryl tylosin (AIV), was developed by Eco Animal Health Products Ltd. in the UK, and its tartrate salt is commonly used. Tylvalosin offers advantages such as high efficacy, low toxicity, and low residue, and does not induce cross-resistance among macrolide antibiotics. It is an effective macrolide antibiotic for the treatment of respiratory and digestive tract infections. It is used to treat mycoplasma infections in pigs and chickens, Brachyspira hyosynospira, and other sensitive bacterial infections, such as swine panting disease, chronic respiratory distress syndrome, blue ear disease, porcine proliferative enteritis, infectious pleuropneumonia, streptococcal disease, and air sacculitis in chickens.

[0003] Chemical structure of Tylvalosin tartrate:

[0004]

[0005] Tylvalosin tartrate Molecular formula: C57H93NO25 Molecular weight: 1192

[0006] Tylvalosin tartrate is a white or off-white powder, which is easily soluble in methanol, soluble in water, acetone or chloroform, slightly soluble in ethyl acetate or ether, and almost insoluble in ethane.

[0007] The soluble powder of tylvalosin tartrate is administered by spray when treating airbag inflammation. If the purity of tylvalosin tartrate is insufficient, it may easily cause the spray dispenser to clog, affecting the efficacy of the drug.

[0008] CN101381756A discloses a method for purifying super tylosin (i.e., tyvalosin) by using acid extraction and alkali precipitation and secondary crystallization. CN102659882A discloses a method for purifying tartrate acetylisovaleryl tylosin (i.e., tyvalosin). Both methods use acid and base displacement technology, have poor selectivity for structural analogs, and cannot effectively remove related substances with similar structures. The final tyvalosin purity is relatively low, reaching only 92.1% at the highest.

[0009] CN108358979B discloses a method for purifying tylvalosin using two organic solvent extractions followed by dissolution crystallization. The organic solvent is selected from at least one of ethyl acetate, butyl acetate, benzene, and toluene, and the dissolution agent is selected from at least one of methyl tert-butyl ether, ethyl ether, and petroleum ether. CN107936074 discloses a method for purifying tylvalosin using organic solvents such as ethyl acetate, isopropyl alcohol, and petroleum ether to recrystallize the crude tylvalosin twice. Both processes utilize organic solvent extraction and recrystallization. While these methods can achieve a tylvalosin purity exceeding 97%, they are complex, result in significant yield losses, and pose numerous safety and environmental risks.

[0010] CN104610403A discloses a method for purifying tylvalosin using silica gel chromatography. This method uses column chromatography silica gel as the separation medium, and can produce a tylvalosin product with a purity exceeding 97%. However, the use of column chromatography results in a complex production process and high production costs, making industrial production difficult.

[0011] CN112409429B discloses a method for obtaining purified tylvalosin using activated carbon adsorption and impurity removal, followed by spray drying of the filtrate. Although this process does not employ an organic solvent, the activated carbon adsorption and impurity removal of 10% (w / w) tylvalosin, followed by spray drying of the filtrate, significantly affects the effectiveness of the activated carbon adsorption and impurity removal, making it difficult to ensure product purity. While the activated carbon adsorbs impurities, it also adsorbs a certain amount of tylvalosin, affecting the decolorization yield of tylvalosin. Once used, the activated carbon cannot be regenerated and reused, and the resulting filter residue becomes solid hazardous waste. Summary of the Invention

[0012] The purpose of the present invention is to overcome the above-mentioned defects of the prior art and provide a method for purifying tyvalosin tartrate.

[0013] The technical solutions adopted to achieve the above objectives are:

[0014] A method for purifying tyvalosin tartrate comprises the following steps:

[0015] (1) dissolving the crude product of tylvalosin, and separating it by adsorption and analysis with a macroporous resin to obtain a tylvalosin tartrate analytical solution;

[0016] (2) decolorizing the tartrate tartrate tantalum solution obtained in step (1) with an anionic resin to obtain a tartrate tartrate decolorized solution;

[0017] (3) The decolorized tartrate tyvalosin solution obtained in step (2) is concentrated by nanofiltration and then spray-dried to obtain pure tartrate tyvalosin.

[0018] Wherein, the weight percentage of the crude tyvalosin in step (1) is greater than or equal to 80%.

[0019] Wherein, the crude tyvalosin product in step (1) is dissolved in 6% (w / w) acidic methanol.

[0020] Wherein, the macroporous resin adsorption feeding rate in step (1) is 1.5 to 1.9 BV / h.

[0021] Wherein, the macroporous resin analysis in step (1) is performed by top separation with water and then analysis with 10% (w / w) tartaric acid.

[0022] Wherein, the analytical feed rate is 0.6-1.2 BV / h.

[0023] Wherein, the tartrate tyvalosin analytical solution in step (1) is a liquid having a collection wavelength of 290 nm and a potency of 100 ug / ml or above.

[0024] Wherein, the macroporous resin in step (1) is a polar macroporous resin;

[0025] Preferably, the macroporous resin is LX-1180 resin

[0026] Wherein, the anion resin decolorization feeding rate in step (2) is 0.8 to 1.2 BV / h.

[0027] Wherein, the anion resin in step (2) is a weakly basic anion resin;

[0028] Preferably, the anion resin is D941 resin.

[0029] Wherein, the concentration multiple of the nanofiltration membrane in step (3) is controlled to be 5 to 7 times.

[0030] The technical solution of the present invention has the following beneficial technical effects:

[0031] The present invention improves the tyvalosin purification process by adopting macroporous resin extraction and anion resin decolorization extraction and purification methods. The process has strong selectivity, high separation purity, high decolorization efficiency, low residue, and the resin can be regenerated and reused. No flammable and explosive organic solvent is used. After purification, the feed liquid is concentrated through a nanofiltration membrane. The entire purification process has strong safety, low production cost, low energy consumption, high product purity, is suitable for large-scale production, and can stably reach a yield of 93.2% and a purity of 98.4%. DETAILED DESCRIPTION

[0032] The present invention is described below with reference to examples. It should be understood that the examples are for the purpose of illustrating the present invention rather than limiting the present invention. The scope and core content of the present invention are determined by reference to the claims.

[0033] The present invention is described below with reference to examples. It should be understood that the examples are for the purpose of illustrating the present invention rather than limiting the present invention. The scope and core content of the present invention are determined by reference to the claims.

[0034] Preparation of 6% (w / w) acidic methanol: Take 75 ml of 95% methanol (density 0.8023 g / ml), add 600 ml of purified water, adjust the pH to 3.0 with 2 mol / L hydrochloric acid, and add purified water to 1000 ml.

[0035] 1400 g of crude tylvalosin (purity 80% (w / w)) was divided into 7 portions and the following experiments were performed on each portion:

[0036] Example 1

[0037] 200g of crude tylvalosin was dissolved in 6% (w / w) acidic methanol with stirring to prepare a 20% (w / w) tylvalosin solution. The solution was fed to a macroporous resin (model LX-1180) at a feed rate of 1.5 BV / h. After completion, the solution was top-washed with water and then resolved with 10% (w / w) tartaric acid at a feed rate of 0.6 BV / h. The collection wavelength was 290 nm. The solution with a titer ≥ 100 μg / ml was obtained to obtain a tylvalosin tartrate solution. The solution was then passed through an anionic resin column (model D941 resin) at a feed rate of 0.8 BV / h to obtain a tylvalosin tartrate decolorized solution. The solution was then passed through a nanofiltration membrane and concentrated to 5 times to obtain a tylvalosin tartrate concentrate. The solution was then spray-dried to obtain pure tylvalosin tartrate. The yield was 92.8% and the purity was 97.9%.

[0038] Example 2

[0039] 200g of crude tylvalosin was dissolved in 6% (w / w) acidic methanol with stirring to prepare a 20% (w / w) tylvalosin solution. The solution was fed to a macroporous resin (model LX-1180) at a feed rate of 1.7 BV / h. After completion, the solution was top-washed with water and then resolved with 10% (w / w) tartaric acid at a feed rate of 1.0 BV / h. The collection wavelength was 290 nm. The solution with a titer ≥ 100 μg / ml was obtained to obtain a tylvalosin tartrate solution. The solution was then passed through an anionic resin column (model D941 resin) at a feed rate of 1.0 BV / h to obtain a tylvalosin tartrate decolorized solution. The solution was then passed through a nanofiltration membrane and concentrated to 6 times to obtain a tylvalosin tartrate concentrate. The solution was spray-dried to obtain pure tylvalosin tartrate. The yield was 93.5% and the purity was 98.1%.

[0040] Example 3

[0041] 200g of crude tylvalosin was dissolved in 6% (w / w) acidic methanol with stirring to prepare a 20% (w / w) tylvalosin solution. The solution was fed to a macroporous resin (model LX-1180) at a feed rate of 1.9 BV / h. After completion, the solution was top-washed with water and then resolved with 10% (w / w) tartaric acid at a feed rate of 1.2 BV / h. The collection wavelength was 290 nm. The solution with a titer ≥ 100 μg / ml was obtained to obtain a tylvalosin tartrate solution. The solution was then passed through an anionic resin column (model D941 resin) at a feed rate of 1.2 BV / h to obtain a tylvalosin tartrate decolorized solution. The solution was then filtered through a nanofiltration membrane and concentrated to 7 times to obtain a tylvalosin tartrate concentrate. The solution was then spray-dried to obtain pure tylvalosin tartrate. The yield was 93.4% and the purity was 98.6%.

[0042] Example 4

[0043] 200g of crude tylvalosin was dissolved in 6% (w / w) acidic methanol with stirring to prepare a 20% (w / w) tylvalosin solution. The solution was fed to a macroporous resin (model LX-1180) at a feed rate of 1.7 BV / h. After completion, the solution was top-washed with water and then resolved with 10% (w / w) tartaric acid at a feed rate of 0.9 BV / h. The collection wavelength was 290 nm. The solution with a titer ≥ 100 μg / ml was obtained to obtain a tylvalosin tartrate solution. The solution was then passed through an anionic resin column (model D941 resin) at a feed rate of 1.0 BV / h to obtain a tylvalosin tartrate decolorized solution. The solution was then passed through a nanofiltration membrane and concentrated to 6 times to obtain a tylvalosin tartrate concentrate. The solution was then spray-dried to obtain pure tylvalosin tartrate. The yield was 92.6% and the purity was 98.8%.

[0044] Example 5

[0045] 200g of crude tylvalosin was dissolved in 6% (w / w) acidic methanol with stirring to prepare a 20% (w / w) tylvalosin solution. The solution was fed to a macroporous resin (model LX-1180) at a feed rate of 1.5 BV / h. After completion, the solution was top-washed with water and then resolved with 10% (w / w) tartaric acid at a feed rate of 1.2 BV / h. The collection wavelength was 290 nm. The solution with a titer ≥ 100 μg / ml was obtained to obtain a tylvalosin tartrate solution. The solution was then passed through an anionic resin column (model D941 resin) at a feed rate of 1.0 BV / h to obtain a tylvalosin tartrate decolorized solution. The solution was then filtered through a nanofiltration membrane and concentrated to 6 times to obtain a tylvalosin tartrate concentrate. The solution was then spray-dried to obtain pure tylvalosin tartrate. The yield was 93.7% and the purity was 98.5%.

[0046] Comparative Case 1

[0047] 200g of crude tylvalosin was dissolved in 6% (w / w) acidic methanol with stirring to produce a 20% (w / w) tylvalosin solution. 12g of activated carbon was added, the temperature maintained at 15-17°C, and stirred for 1 hour. The activated carbon was then filtered to remove the decolorized filtrate, which was spray-dried to obtain pure tylvalosin tartrate. The yield was 94.3% and the purity was 86.5%.

[0048] Comparative Case 2

[0049] 200g of crude tylvalosin was dissolved in 6% (w / w) acidic methanol with stirring to prepare a 20% (w / w) tylvalosin solution. The solution was fed to a macroporous resin (model LX-1180) at a feed rate of 1.9 BV / h. After completion, the solution was top-washed with water and then resolved with 10% (w / w) tartaric acid at a feed rate of 1.3 BV / h. The collection wavelength was 290 nm. The solution with a titer ≥ 100 μg / ml was obtained to obtain a tylvalosin tartrate solution. The solution was then passed through an anionic resin column (model D941 resin) at a feed rate of 1.3 BV / h to obtain a tylvalosin tartrate decolorized solution. The solution was then filtered through a nanofiltration membrane and concentrated to 4 times to obtain a tylvalosin tartrate concentrate. The solution was then spray-dried to obtain pure tylvalosin tartrate. The yield was 90.7% and the purity was 88.8%.

[0050] Comparative Case 3

[0051] 200g of crude tylvalosin was dissolved in 6% (w / w) acidic methanol with stirring to prepare a 20% (w / w) tylvalosin solution. The solution was fed to a macroporous resin (model LX-1180) at a feed rate of 1.9 BV / h. After completion, the solution was top-washed with water and then resolved with 10% (w / w) tartaric acid at a feed rate of 1.0 BV / h. The solution was collected at a wavelength of 290 nm. The solution containing a titer of ≥100 μg / ml was filtered through a nanofiltration membrane and concentrated to 6 times to obtain a tylvalosin tartrate concentrate. This concentrate was then spray-dried to obtain pure tylvalosin tartrate. The yield was 91.1% and the purity was 86.2%.

[0052] Effect comparison:

[0053]

Claims

1. A method for purifying tyvalosin tartrate, comprising the following steps: (1) dissolving the crude product of tylvalosin, and separating it by adsorption and analysis with a macroporous resin to obtain a tylvalosin tartrate analytical solution; (2) decolorizing the tartrate tartrate tantalum solution obtained in step (1) with an anionic resin to obtain a tartrate tartrate decolorized solution; (3) The decolorized tartrate tyvalosin solution obtained in step (2) is concentrated by nanofiltration and then spray-dried to obtain pure tartrate tyvalosin.

2. The purification method according to claim 1, wherein The weight percentage of the crude tyvalosin in step (1) is greater than or equal to 80%.

3. The purification method according to claim 1, wherein The crude tyvalosin product in step (1) is dissolved in 6% (w / w) acidic methanol.

4. The purification method according to claim 1, wherein The macroporous resin adsorption feeding rate in step (1) is 1.5 to 1.9 BV / h.

5. The purification method according to claim 1, wherein The macroporous resin analysis in step (1) is performed by top separation with water and then analysis with 10% (w / w) tartaric acid.

6. The purification method according to claim 5, characterized in that The analytical feed rate is 0.6 to 1.2 BV / h.

7. The purification method according to claim 1, characterized in that The tartrate tyvalosin analytical solution in step (1) is a liquid having a collection wavelength of 290 nm and a potency of 100 ug / ml or above.

8. The purification method according to claim 1, characterized in that The macroporous resin in step (1) is a polar macroporous resin; Preferably, the macroporous resin is LX-1180 resin.

9. The purification method according to claim 1, characterized in that The anion resin decolorization feeding rate in step (2) is 0.8 to 1.2 BV / h.

10. The purification method according to claim 1, characterized in that The anion resin in step (2) is a weakly basic anion resin; Preferably, the anion resin is D941 resin.

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