Method for extracting tylosin tartrate

Abstract

The invention relates to a method for refining tylosin tartrate, which comprises the following steps of: dehydrating tylosin extract liquor by using a 4A molecular sieve, decoloring by using activated carbon, and finally feeding a tartaric acid-butanol saturated solution to crystallize to obtain a finished product of the tylosin tartrate. According to the method, the tylosin tartrate is extracted by combining a molecular sieve dehydration method with a tartaric acid-butanol saturated solution fed-batch crystallization method, so that the extraction process is simplified, the purity and the yield of the tylosin tartrate product are effectively improved, and industrial production is easy to realize.

Description

Technical Field

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

[0002] Tylosin, also known as tylosin and tylosin, is an important macrolide antibiotic for livestock and poultry. Tylosin tartrate is the tartrate salt of tylosin, with a molecular formula of 2(C 46 H 77 NO 17 )·C4H6O6, molecular weight 1982.31, is produced by Streptomyces freundii as white or light yellow powder, a white plate-like crystal, slightly soluble in water, and the aqueous solution is alkaline; Tylosin tartrate is mainly used in the clinic to treat and prevent various respiratory, intestinal, reproductive and locomotor system infections caused by pathogens such as mycoplasma, Staphylococcus aureus, Bacillus pyogenes, Pneumococcus, Erysipelothrix, Haemophilus parahaemolyticus, Neisseria meningitidis, Pasteurella, spirochetes, cocci and other pathogens. Under the condition of pH 3.7-9.5, the tylosin solution is very stable: when pH < 3.7, tylosin is decomposed into desmycosin and mycosyl; when pH > 9.5, the lactone ring of tylosin is easily destroyed.

[0003] The domestic tylosin extraction process mainly pre-treats the tylosin fermentation liquid, separates the solid from the liquid, and obtains the fermentation filtrate; secondly, the fermentation filtrate is subjected to liquid-liquid extraction with an organic solvent, tylosin enters the ester phase liquid, the ester phase liquid is acidified and stripped, and tylosin enters the aqueous phase; finally, the tylosin aqueous phase liquid is neutralized, decolorized, filtered, and spray-dried to obtain the tylosin tartrate finished product. This method has the following problems: ① It is not easy to accurately control the pH during the extraction process, wherein when the pH is too high, tylosin is easy to precipitate, resulting in tylosin loss, and when the pH is too low, tylosin tartrate is easy to degrade; ② During spray drying, it is decomposed and destroyed due to the long heating time, and more than 5% of tylosin is lost, thereby affecting the quality and yield of the finished product. Summary of the invention

[0004] The invention provides a method for refining tylosin tartrate. Tylosin tartrate is refined by 4A molecular sieve dehydration treatment combined with tartaric acid-butanol saturated solution feed crystallization method, so that the purity and yield of the tylosin tartrate product are effectively improved while simplifying the extraction process, and industrial production is easily realized.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is:

[0006] A method for extracting tylosin tartrate comprises the following steps: firstly dehydrating the tylosin extract through a 4A molecular sieve, then decolorizing through activated carbon, and finally adding a tartaric acid-butanol saturated solution for crystallization to obtain a tylosin tartrate finished product.

[0007] Wherein, the potency of the tylosin extract is greater than 18000u / ml.

[0008] Wherein, the addition amount of the 4A molecular sieve is 3-5:50 (W / V);

[0009] Before use, the 4A molecular sieve needs to be spread flat on an enamel tray and baked at high temperature. The oven temperature is set at 260-280°C and the baking time is 4-6 hours. Remove deep water and activate.

[0010] The water content of the tylosin extract after dehydration is less than or equal to 0.6%.

[0011] Wherein, the added amount of the activated carbon is 2-4% (W / V).

[0012] Wherein, the addition amount of the tartaric acid-butanol saturated solution is 2.5-3.0% (V / V).

[0013] The tartaric acid-butanol saturated solution is prepared by heating a butanol water bath to 70° C. and adding tartaric acid.

[0014] Wherein, the ratio of tartaric acid to butanol is 70 / 500 (V / V).

[0015] The technical solution of the present invention has the following beneficial technical effects:

[0016] 1. The present invention achieves the purpose of reducing the water content of the extract by dehydrating the 4A molecular sieve. Experiments show that the water content of the extract is lower than 0.6%, which is more conducive to the crystallization of saturated tartaric acid-butanol in the subsequent flow and effectively avoids the occurrence of emulsification in the subsequent process.

[0017] 2. The saturated tartaric acid-butanol crystallization method is adopted to avoid the pH being too high or too low in the extraction process of tylosin tartrate, and effectively improve the extraction yield of tylosin tartrate;

[0018] 3. The present invention eliminates the spray drying step and simplifies the extraction process by combining the dehydration of the molecular sieve with the saturated tartaric acid-butanol crystallization method. The yield of the tylosin tartrate product is more than 90%, while the A component reaches more than 93% and the total component reaches more than 97%, effectively improving the product yield and product quality. DETAILED DESCRIPTION

[0019] The embodiments of the present invention will be described clearly and completely below in conjunction with the examples. Obviously, the described examples are only some examples of the present invention, rather than all examples. The following description of at least one exemplary embodiment is actually only illustrative and is by no means intended to limit the present invention and its application or use. Based on the examples in the present invention, all other examples obtained by ordinary technicians in the field without creative work are within the scope of protection of the present invention.

[0020] The tylosin extract in the following examples and comparative examples was prepared by the following method: the tylosin fermentation broth was first adjusted to acidity with 28% aluminum sulfate and then adjusted to alkalinity with 30% liquid alkali, and the filtrate obtained after solid-liquid separation was extracted with butyl acetate to obtain a tylosin extract with a potency of 24500u / ml.

[0021] Example 1

[0022] Take 50L of tylosin extract, the potency is 24500u / ml;

[0023] Extraction liquid removal: The tylosin extract was allowed to stand for 30 min, the impurities in the lower layer were removed, and the water content was measured to be 1.02%.

[0024] Molecular sieve drying: Spread the 4A molecular sieve flat on an enamel tray, put it into an oven, turn on the oven and set it to 260°C for 6 hours.

[0025] Dehydration: Weigh 4 kg of 4A molecular sieve and add it to the extract, rotate at 60 r / min, stir for 30 minutes, and measure the water content to be 0.60%.

[0026] Decolorization: Add 1.5 kg of activated carbon to the extract, stir for 30 min, rotate at 60 r / min, and filter.

[0027] Reagent preparation: Prepare a saturated solution of tartaric acid-butanol at 70°C.

[0028] Crystallization: at 40°C, add 1.225 L of saturated tartaric acid-butanol solution. After the addition, continue stirring for 30 min at a stirring speed of 60 r / min and let stand for 30 min.

[0029] Drying: The precipitated crystals were dried in an oven at 80° C. The Tylos tartrate product was obtained with a yield of 90.2%, 94.1% of component A, and 97.8% of total components.

[0030] Example 2

[0031] Take 50L of tylosin extract, the potency is 24500u / ml;

[0032] Extraction liquid removal: The tylosin extract was allowed to stand for 30 min, the impurities in the lower layer were removed, and the water content was measured to be 1.14%.

[0033] Molecular sieve drying: Spread the 4A molecular sieve flat on an enamel tray, put it into an oven, turn on the oven and set it to 265°C for 5.5 hours.

[0034] Dehydration: Weigh 5 kg of 4A molecular sieve and add it to the extract, rotate at 60 r / min, stir for 30 min, and the water content is 0.41%.

[0035] Decolorization: Add 1.47 kg of activated carbon to the extract, stir for 30 min, rotate at 60 r / min, and filter.

[0036] Reagent preparation: Prepare a saturated solution of tartaric acid-butanol at 70°C.

[0037] Crystallization: at 40°C, add 1.341 L of saturated tartaric acid-butanol solution. After the addition, continue stirring for 30 min at a stirring speed of 60 r / min and let stand for 30 min.

[0038] Drying: The precipitated crystals were dried in an oven at 80° C. The Tylos tartrate product was obtained with a yield of 90.7%, 94.3% of component A, and 97.6% of total components.

[0039] Example 3

[0040] Take 50L of tylosin extract, the potency is 24500u / ml;

[0041] Extraction liquid removal: The tylosin extract was allowed to stand for 30 min, the impurities in the lower layer were removed, and the water content was measured to be 1.25%.

[0042] Molecular sieve drying: Spread the 4A molecular sieve flat on an enamel tray, put it into an oven, turn on the oven and set it to 270°C for 5.3 hours.

[0043] Dehydration: Weigh 3kg 4A molecular sieve and add it to the extract, stir for 30min, rotation speed 60r / min, water content 0.48%.

[0044] Decolorization: Add 1.65 kg of activated carbon to the extract, stir for 30 min, rotate at 60 r / min, and filter.

[0045] Reagent preparation: Prepare a saturated solution of tartaric acid-butanol at 70°C.

[0046] Crystallization: at 40°C, add 1.278 L of saturated tartaric acid-butanol solution. After the addition, continue stirring for 30 min at a stirring speed of 60 r / min and let stand for 30 min.

[0047] Drying: The precipitated crystals were dried in an oven at 80° C. The Tylos tartrate product was obtained with a yield of 90.2%, 93.7% of component A, and 97.8% of total components.

[0048] Example 4

[0049] Take 50L of tylosin extract, the potency is 24500u / ml;

[0050] Extraction liquid removal: Take the tylosin extract, let it stand for 30 minutes, remove the impurities in the lower layer, and measure the water content to be 1.16%.

[0051] Molecular sieve drying: Spread the 4A molecular sieve flat on an enamel tray, put it into an oven, turn on the oven and set it to 280℃ for 4 hours.

[0052] Dehydration: Weigh 4.5 kg of 4A molecular sieve and add it to the extract, stir for 30 minutes, set at 60 r / min, and the water content is 0.39%.

[0053] Decolorization: Add 1.47 kg of activated carbon to the extract, stir for 30 min, set to 60 r / min, and filter.

[0054] Reagent preparation: Prepare a saturated solution of tartaric acid-butanol at 70°C.

[0055] Crystallization: at 40°C, add 1.306 L of saturated tartaric acid-butanol solution. After the addition, continue stirring for 30 min at a stirring speed of 60 r / min and let stand for 30 min.

[0056] Drying: The precipitated crystals were dried in an oven at 80° C. The Tylos tartrate product was obtained with a yield of 90.6%, 93.8% of component A, and 97.4% of total components.

[0057] Comparative Example 1

[0058] Take 50L of tylosin extract, the potency is 24500u / ml;

[0059] Extraction liquid removal: Take the tylosin extract, let it stand for 30 minutes, remove the impurities in the lower layer, and measure the water content to be 1.17%.

[0060] Decolorization: Add 1.48 kg of activated carbon to the extract, stir for 30 min, set to 60 r / min, and filter.

[0061] Reagent preparation: Prepare a saturated solution of tartaric acid-butanol at 70°C.

[0062] Crystallization: at 40°C, add 1.247 L of saturated tartaric acid-butanol solution. After the addition, continue stirring for 30 min at a stirring speed of 60 r / min and let stand for 30 min.

[0063] Drying: The precipitated crystals were dried in an oven at 80° C. The Tylos tartrate product was obtained with a yield of 89.1%, 89.2% of component A, and 93.5% of total components.

[0064] Comparative Example 2

[0065] Take 50L of tylosin extract, the potency is 24500u / ml;

[0066] Extraction liquid removal: Take the tylosin extract, let it stand for 30 minutes, remove the impurities in the lower layer, and measure the water content to be 1.37%.

[0067] Molecular sieve drying: Spread the 4A molecular sieve flat on an enamel tray, put it into an oven, turn on the oven and set it to 270°C for 6 hours.

[0068] Dehydration: Weigh 4 kg of 4A molecular sieve and add it to the extract, stir for 30 minutes, speed 60 r / min, water content 0.41%.

[0069] Crystallization decolorization: The temperature of the extract after dehydration is reduced to 12°C, and it is acidified with 4% tartaric acid and allowed to stand to obtain a stripping solution, which is then neutralized with 23% calcium hydroxide to obtain a refined solution. 1.52 kg of activated carbon is added to the refined solution, the speed is 60 r / min, and the mixture is stirred for 30 minutes before filtering.

[0070] Drying: The refined liquid was spray dried by spray drying to obtain the finished product of tylosin tartrate, with a yield of 87.8%, component A of 92.7%, and total components of 95.5%.

[0071] Comparative Example 3

[0072] Take 50L of tylosin extract, the potency is 24500u / ml;

[0073] Extraction liquid removal: Take the tylosin extract, let it stand for 30 minutes, remove the impurities in the lower layer, and measure the water content to be 1.09%.

[0074] Molecular sieve drying: Spread the 4A molecular sieve flat on an enamel tray, put it into an oven, turn on the oven and set it to 270°C for 5 hours.

[0075] Dehydration: Weigh 4.2 kg of 4A molecular sieve and add it to the extract, rotate at 60 r / min, stir for 30 min, and measure the water content to be 0.39%.

[0076] Decolorization: Add 1.47 kg of activated carbon to the extract, stir for 30 min, rotate at 60 r / min, and filter.

[0077] Reagent preparation: Prepare a saturated solution of tartaric acid-butanol at 70°C.

[0078] Crystallization: at 40°C, add 1.254 L of saturated tartaric acid-butanol solution. After the addition, continue stirring for 30 min at a stirring speed of 60 r / min and let stand for 30 min.

[0079] Drying: The precipitated crystals were dried in an oven at 80° C. The Tylos tartrate product was obtained with a yield of 90.6%, 93.8% of component A, and 97.4% of total components.

Claims

1. A method for extracting tylosin tartrate, comprising the following steps: The tylosin extract was first dehydrated by 4A molecular sieve, then decolorized by activated carbon, and finally crystallized by adding tartaric acid-butanol saturated solution to obtain the tylosin tartrate finished product.

2. The method for extracting tylosin tartrate according to claim 1, characterized in that The potency of the tylosin extract is greater than 18000u / ml.

3. The method for extracting tylosin tartrate according to claim 1, characterized in that The addition amount of the 4A molecular sieve is 3-5:50 (W / V).

4. The method for extracting tylosin tartrate according to claim 1, characterized in that The water content of the tylosin extract after dehydration is less than or equal to 0.6%.

5. The method for extracting tylosin tartrate according to claim 1, characterized in that The added amount of the activated carbon is 2-4% (W / V).

6. The method for extracting tylosin tartrate according to claim 1, characterized in that The addition amount of the tartaric acid-butanol saturated solution is 2.5-3.0% (V / V).

7. The method for extracting tylosin tartrate according to claim 6, characterized in that The tartaric acid-butanol saturated solution is prepared by heating butanol in a water bath to 70° C. and adding tartaric acid.

8. The method for extracting tylosin tartrate according to claim 6, characterized in that The ratio of tartaric acid to butanol is 70 / 500 (V / V).