阅读说明：本技术 一种通过离子交换树脂分离纯化夫西地酸的方法 (method for separating and purifying fusidic acid through ion exchange resin ) 是由 张茂华 蒋永飞 赖庆云 陈亮 吴耀美 于 2019-09-19 设计创作，主要内容包括：本发明公开了一种通过离子交换树脂分离纯化夫西地酸的方法,利用夫西地酸在pH值大于等于电点的条件下水溶性增大和离子交换树脂的强选择性吸附这两个特点,采用发酵液经过滤后得澄清的夫西地酸水溶液,发酵滤液经离子交换树脂吸附解析,最后将解析液经浓缩后用有机溶剂萃取并结晶烘干,得到符合质量要求的夫西地酸成品,解决了目前大多数生产工艺工序步骤繁琐,生产周期长,有机溶剂使用量较大等问题,本发明的整体工艺中工序步骤少,生产周期短,生产效率高,同时有机溶剂用量少,提高了生产安全性。(The invention discloses a method for separating and purifying fusidic acid by ion exchange resin, which utilizes the two characteristics of increased water solubility and strong selective adsorption of the ion exchange resin of the fusidic acid under the condition that the pH value is more than or equal to an electric point, adopts clear fusidic acid aqueous solution obtained by filtering fermentation liquor, obtains a fusidic acid finished product meeting the quality requirement by adsorbing and resolving the fermentation filtrate by the ion exchange resin, and finally extracts, crystallizes and dries the resolved solution by using an organic solvent after concentrating, thereby solving the problems of complicated steps, long production period, large usage amount of the organic solvent and the like of most of the conventional production process.)

1. A process for the separation and purification of fusidic acid by means of an ion exchange resin, characterised in that: the method comprises the following steps:

(1) Selecting and filtering fusidic acid fermentation liquor filtering equipment;

(2) Adsorbing the filtered filtrate by using ion exchange resin, wherein the content of the adsorption waste liquid is less than 100mg/L, the adsorption capacity of the resin is about 30g/L, after adsorption saturation, using 0.7mol of sodium chloride aqueous solution to analyze the saturated resin, detecting the content of the analysis liquid by using liquid chromatography, and collecting part of analysis liquid with the content of more than 500 mg/L;

(3) Concentrating the analytic solution to the concentration of 50-60 g/l, controlling the concentration temperature at 60 ℃, extracting the concentrated solution by using ethyl acetate with the volume of 1 time, adjusting the pH value to 5.0-5.5 by using dilute hydrochloric acid, standing for layering, and collecting an extracted lipid phase;

(4) concentrating the extracted lipid phase by single effect, controlling the concentration temperature at 50-55 ℃, concentrating to the concentration of 220-250 g/l, cooling, stirring, crystallizing for 12 hours, and filtering to obtain crude fusidic acid;

(5) refining the crude fusidic acid with ethyl acetate, and drying to obtain a finished fusidic acid product with the purity of more than 99.5%.

2. the method for separating and purifying fusidic acid through ion exchange resin according to claim 1, wherein fusidic acid fermentation liquor in step (1) is filtered by using a plate-and-frame filter, a filter aid with the volume of 1-2% is added into the fusidic acid fermentation liquor, the mixture is stirred for 10-30 minutes, then the mixture is filtered by using the plate-and-frame filter, and filter cakes are washed by drinking water after the filtration.

3. A process for separation and purification of fusidic acid by ion exchange resin according to claim 1, wherein in step (1) fusidic acid fermentation broth is selected from membrane filtration, and membrane filtration equipment is selected from ultrafiltration membrane or hollow fiber membrane.

4. A process for the separation and purification of fusidic acid by ion exchange resin according to claim 1, wherein in step (2) the ion exchange resin may be selected from KT-30, D303 or D315 ion exchange resin.

5. A process for separation and purification of fusidic acid via ion exchange resin according to claim 1, wherein in step (3) the eluent concentration is selected from membrane concentration or climbing-film concentration.

6. A process for the separation and purification of fusidic acid by ion exchange resin according to claim 1, wherein the extraction solvent in step (3) is selected from the group consisting of the lipid solvents ethyl acetate or isopropyl acetate.

the technical field is as follows:

the invention belongs to the technical field of chemical substance extraction, and particularly relates to a method for separating and purifying fusidic acid through ion exchange resin.

Background art:

fusidic acid belongs to fusidic acid antibiotics, and is found and extracted from a fusidic acid lipococcus fermentation broth by Danish Liao pharmaceutical company in 1962. Fusidic acid is a water-insoluble colorless crystal that is soluble in organic solvents such as alcohols and lipids. Fusidic acid is a weak acid with a pKa of 5.7. The sodium salt form is generally used clinically and is easy to dissolve in water.

A method for extracting and separating fusidic acid, which is disclosed in a patent of 'a method for extracting and separating fusidic acid' published by Jiuyangyang biopharmaceutical Limited in Jiangxi of 2010, is characterized in that fermentation liquor is subjected to membrane filtration, then is adsorbed by macroporous resins D4020 and D312, is resolved by ethanol, and is heated, decompressed and concentrated, and then is extracted by a lipid organic solvent and is crystallized. The scheme has the advantages of more organic solvents, large consumption of resolved ethanol, large amount of steam required during ethanol recovery, high energy consumption and high cost.

The separation and purification method mentioned in the preliminary research on the fermentation and separation and purification process of fusidic acid, which is a paper of Master university in Shanghai in 2012, is to obtain mycelium containing fusidic acid by centrifugally filtering fermentation liquid under acidic condition, leaching the mycelium with ethanol or methanol after drying the mycelium, adsorbing and analyzing the leaching solution by macroporous resin, and concentrating the analyzing solution under reduced pressure to obtain fusidic acid white powder with the purity of more than 90%. Although the process method is simple, the solvent consumption is low, the purity of the obtained product is poor, and the market demand cannot be met.

At present, the existing production process mostly uses fermentation liquor to acidify and filter to obtain mycelium, the mycelium is dried and then uses alcohol organic solvent to extract dry mycelium, and the extract liquor is concentrated to remove alcohol, then uses lipid organic solvent to extract and crystallize. The method has the advantages of large alcohol amount for mycelium extraction, low production safety, large energy consumption during recovery and high cost although the alcohol can be recovered and reused.

in the process of Jiangxi Jiuyangyang biological pharmacy Co., Ltd, although fusidic acid is transferred to a water phase by a membrane filtration method, the processes of mycelium drying and alcohol extraction are reduced, and the ethanol dosage in the step is solved, a large amount of ethanol is still used when macroporous adsorption resin is used for adsorption and resolution in the subsequent process, the solvent dosage is not improved on the whole, the production safety is not improved, and the energy consumption cost is not reduced.

the invention content is as follows:

in view of the above problems, the technical problem to be solved by the present invention is to provide a process for the separation and purification of fusidic acid by ion exchange resin.

The invention relates to a method for separating and purifying fusidic acid by using ion exchange resin, which comprises the following steps:

(1) selecting and filtering fusidic acid fermentation liquor filtering equipment;

(2) Adsorbing the filtered filtrate by using ion exchange resin, wherein the content of the adsorption waste liquid is less than 100mg/L, the adsorption capacity of the resin is about 30g/L, after adsorption saturation, using 0.7mol of sodium chloride aqueous solution to analyze the saturated resin, detecting the content of the analysis liquid by using liquid chromatography, and collecting part of analysis liquid with the content of more than 500 mg/L;

(3) Concentrating the analysis solution to the concentration of 50-60 g/l, controlling the concentration temperature at 60 ℃, extracting the concentrated solution by using ethyl acetate with the volume of 1 time that of the concentrated solution, adjusting the pH value to be less than 5.7 by using dilute hydrochloric acid, standing for layering, and collecting an extracted lipid phase;

(4) concentrating the extracted lipid phase by single effect, cooling, stirring, crystallizing for 12 hours, and filtering to obtain crude fusidic acid;

(5) Refining the crude fusidic acid with ethyl acetate, and drying to obtain a finished fusidic acid product with the purity of more than 99.5%.

preferably, the fusidic acid fermentation liquor in the step (1) is filtered by using a plate-and-frame filter, a filter aid with the volume of 1-2% is added into the fusidic acid fermentation liquor, the mixture is stirred for 10-30 minutes, and then the mixture is pumped into the plate-and-frame filter, and after the filtration, the filter cake is washed by drinking water.

Preferably, the fusidic acid fermentation broth in the step (1) is filtered by a membrane, and the membrane filtering equipment can be an ultrafiltration membrane or a hollow fiber membrane.

Preferably, the ion exchange resin in the step (2) can be KT-30, D303 or D315 ion exchange resin.

preferably, the concentration of the solution in the step (3) may be membrane concentration or climbing-film concentration.

Preferably, the extraction solvent in step (3) may be selected from ethyl acetate or isopropyl acetate.

The invention has the beneficial effects that: the invention adopts the process route of fermentation liquor → filtration → ion exchange resin adsorption → salt solution analysis → concentration → extraction → crystallization, and utilizes the two characteristics of water solubility increase of the fusidic acid under the condition that the pH value is more than or equal to the electric point and strong selective adsorption of the ion exchange resin, the fermentation liquor is filtered to obtain clear fusidic acid aqueous solution, the fermentation filtrate is absorbed and analyzed by the ion exchange resin, and finally the analyzed solution is concentrated, extracted by organic solvent, crystallized and dried to obtain the fusidic acid finished product meeting the quality requirement. The problems that most of the existing production processes are complex in process steps, long in production period, large in organic solvent usage amount and the like are solved, the overall process is few in process steps, short in production period, high in production efficiency, small in organic solvent usage amount and capable of improving production safety.

Description of the drawings:

FIG. 1 is a process flow diagram of example 1 of the present invention;

FIG. 2 is a process flow diagram of example 2 of the present invention

FIG. 3 is a process flow diagram of example 3 of the present invention.

The specific implementation mode is as follows:

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described below by way of specific embodiments and accompanying drawings. It is to be understood that such description is merely illustrative and not intended to limit the scope of the present invention. Moreover, in the following description, descriptions of well-known structures and techniques are omitted so as to not unnecessarily obscure the concepts of the present invention.

The method for separating and purifying fusidic acid by using ion exchange resin comprises the following steps:

(1) Selecting and filtering fusidic acid fermentation liquor filtering equipment;

(2) Adsorbing the filtered filtrate by using ion exchange resin, wherein the content of the adsorption waste liquid is less than 100mg/L, the adsorption capacity of the resin is about 30g/L, after adsorption saturation, using 0.7mol of sodium chloride aqueous solution to analyze the saturated resin, detecting the content of the analysis liquid by using liquid chromatography, and collecting part of analysis liquid with the content of more than 500 mg/L;

(3) Concentrating the analysis solution to the concentration of 50-60 g/l, controlling the concentration temperature at 60 ℃, extracting the concentrated solution by using ethyl acetate with the volume of 1 time that of the concentrated solution, adjusting the pH value to be less than 5.7 by using dilute hydrochloric acid, standing for layering, and collecting an extracted lipid phase;

(4) Concentrating the extracted lipid phase by single effect, cooling, stirring, crystallizing for 12 hours, and filtering to obtain crude fusidic acid;

(5) refining the crude fusidic acid with ethyl acetate, and drying to obtain a finished fusidic acid product with the purity of more than 99.5%.

Specifically, filtering the fusidic acid fermentation liquor in the step (1) by using a plate-and-frame filter, adding a filter aid with the volume of 1-2% into the fusidic acid fermentation liquor, stirring for 10-30 minutes, then pumping into the plate-and-frame filter, and washing a filter cake by using drinking water after filtering; in the step (1), the fusidic acid fermentation liquor is filtered by a membrane, and ultrafiltration membranes or hollow fiber membranes can be used as membrane filtering equipment. The ion exchange resin in the step (2) can be KT-30, D303 or D315 ion exchange resin. In the step (3), the concentration of the analytic solution can be membrane concentration or climbing membrane concentration, and the extraction solvent can be lipid solvent ethyl acetate or isopropyl acetate.