阅读说明：本技术 一种降低林可霉素b组分含量的纯化工艺 (Purification process for reducing content of lincomycin B component ) 是由 吴海波 周永正 梁新建 于 2018-07-10 设计创作，主要内容包括：本发明公开了一种降低林可霉素B组分含量的纯化工艺。该工艺以溶媒提取液的反萃液或林可霉素粗品的水溶液为纯化对象,将溶液用NaOH调pH后在硅胶基质填料的色谱柱上进行吸附,以纯水进行洗涤,再用丁醇或丙酮进行解析,之后用盐酸调酸、结晶。本纯化工艺使用了高效制备液相色谱技术,具有分离收率高,分离周期短,工艺稳定性好,自动化程度高的优势。相对其它纯化工艺,该工艺设备占地少,环境污染小,生产可控性强,有利于工业规模应用。(The invention discloses a purification process for reducing the content of a lincomycin B component. The process uses a back extraction solution of a solvent extraction solution or an aqueous solution of a lincomycin crude product as a purification object, adjusts the pH of the solution by NaOH, then adsorbs the solution on a chromatographic column filled with silica gel matrix, washes the solution by pure water, then resolves the solution by butanol or acetone, and then adjusts the acid by hydrochloric acid and crystallizes the solution. The purification process uses a high performance preparative liquid chromatography technology, and has the advantages of high separation yield, short separation period, good process stability and high automation degree. Compared with other purification processes, the process equipment occupies less land, has little environmental pollution and strong production controllability, and is beneficial to industrial scale application.)

1. A purification process for reducing the content of a lincomycin B component is characterized by comprising the following steps: using the back extraction solution of the solvent extract or the aqueous solution of the lincomycin crude product as a purification object, adjusting the pH of the solution by NaOH, adsorbing the solution on a chromatographic column filled with silica gel matrix, washing the solution by pure water, resolving the solution by butanol or acetone, adjusting the acid by hydrochloric acid, and crystallizing the solution.

2. A purification process for reducing the content of lincomycin B component according to claim 1, wherein: the silica gel matrix filler is filled into a high-pressure chromatographic column or a medium-low pressure column according to the particle size.

3. A purification process for reducing the content of lincomycin B component according to claim 1, wherein: and (4) replacing the back extraction solution or the aqueous solution of the crude product with the fermentation filtrate or the aqueous solution of the recovered crude product for purification and separation.

4. A purification process for reducing the content of lincomycin B component according to claim 1, wherein: the solution was adjusted to a pH of between 7.5 and 8.5 with NaOH and then adsorbed on a silica gel-based packed chromatographic column.

5. A purification process for reducing the content of lincomycin B component according to claim 1, wherein: the sample loading mass of the lincomycin is controlled to be 7-13% of the mass of the filler.

6. The purification process for reducing the content of the lincomycin B component according to claim 1, which specifically comprises the following steps:

① adjusting pH of the back extract of the solvent extract or the aqueous solution of the lincomycin crude product to 7.5-8.5 with NaOH to obtain alkaline solution;

②, pumping the alkalines into a chromatographic column filled with silica gel matrix filler, and controlling the sample loading mass of lincomycin to be 7-13% of the filler mass;

③ washing the column with 0.5-2BV pure water, collecting the fraction by stages, and considering whether the fraction is used indiscriminately according to the content of component B after HPLC detection;

④ is resolved with 1.5-6BV acetone containing 0-10% water or butanol containing 0-20% water;

⑤ adding hydrochloric acid to adjust pH to below 2, directly crystallizing acetone solution, concentrating butanol solution, crystallizing or back extracting with acid water, and crystallizing with acetone;

⑥ the column was equilibrated with 2-5BV of pure water and the process could be repeated ① to ⑤.

Technical Field

The invention belongs to the technical field of pharmaceutical chemicals, and particularly relates to a purification process for reducing the content of a lincomycin B component.

Background

Lincomycin produces a certain amount of lincomycin B in the fermentation process. Because the component B has lower antibacterial activity and higher toxicity, the requirements on the component B are increasingly strict due to lincomycin and derivative products of the lincomycin, clindamycin phosphate and the like. The current means for reducing the content of the component B mainly comprise: 1. after the higher alcohol extraction, the component B is washed with alkaline water. The method has poor controllability, large yield loss and large amount of waste water. 2. And separating the hydrochloric acid back extraction solution or the crude product water solution by resin to remove the component B. The method has the advantages of long period, low efficiency and non-centralized analysis unit, and a large amount of wastewater can be generated after the resin is regenerated by acid and alkali. Therefore, a more efficient purification process for reducing the content of the lincomycin B component is needed.

Disclosure of Invention

In order to overcome some defects of the traditional purification process, the invention develops a high-efficiency purification process for reducing the content of the lincomycin B component.

The technical scheme of the invention is as follows:

a purification process for reducing the content of a lincomycin B component is characterized by comprising the following steps: using the back extraction solution of the solvent extract or the aqueous solution of the lincomycin crude product as a purification object, adjusting the pH of the solution by NaOH, adsorbing the solution on a chromatographic column filled with silica gel matrix, washing the solution by pure water, resolving the solution by butanol or acetone, adjusting the acid by hydrochloric acid, and crystallizing the solution.

The separation column of the invention can be filled into a high-pressure chromatographic column or a medium-low pressure column according to the particle size of the filler. Meanwhile, the fermentation filtrate or the water solution for recovering the crude product can be used for replacing the back extraction solution or the water solution for the crude product for purification and separation.

The purification process for reducing the content of the lincomycin B component is characterized by comprising the following steps of: adjusting pH of the solution to 7.5-8.5 with NaOH, and adsorbing on a chromatographic column filled with silica gel matrix; the sample loading mass of the lincomycin is controlled to be 7-13% of the mass of the filler.

The purification process for reducing the content of the lincomycin B component comprises the following steps and operations:

① adjusting pH of the back extract of the solvent extract or the aqueous solution of the lincomycin crude product to 7.5-8.5 with NaOH to obtain alkaline solution;

②, pumping the alkalines into a chromatographic column filled with silica gel matrix filler, and controlling the sample loading mass of lincomycin to be 7-13% of the filler mass;

③ washing the column with 0.5-2BV pure water, collecting the fraction by stages, and considering whether the fraction is used indiscriminately according to the content of component B after HPLC detection;

④ is resolved with 1.5-6BV acetone containing 0-10% water or butanol containing 0-20% water;

⑤ adding hydrochloric acid to adjust pH to below 2, directly crystallizing acetone solution, concentrating butanol solution, crystallizing or back extracting with acid water, and crystallizing with acetone;

⑥ the column was equilibrated with 2-5BV of pure water and the process could be repeated ① to ⑤.

The invention takes the high performance preparative liquid chromatography as a purification means, and has the advantages of high separation yield, short separation period, good process stability and high automation degree. Compared with other purification processes, the process equipment occupies less land, has little environmental pollution and strong production controllability, and is beneficial to industrial scale application.

Drawings

FIG. 1 example 1 UV monitoring spectrum (210 nm);

FIG. 2 example 2 UV monitoring spectrum (210 nm);

FIG. 3 example 3 UV monitoring spectrum (210 nm);

FIG. 4 chromatographic assay of the feed solution before purification in example 3;

FIG. 5 chromatographic detection of the product obtained in example 3;

FIG. 6 example 4 UV monitoring spectrum (210 nm);

FIG. 7 example 5 UV monitoring spectrum (210 nm);

FIG. 8 example 6 UV monitoring spectrum (210 nm);

FIG. 9 example 7 UV monitoring spectrum (210 nm);

FIG. 10 UV monitoring spectrum (210nm) of example 8.

Detailed Description

The following embodiment is used to specifically describe a high-efficiency lincomycin purification process of the present invention.