庆大霉素发酵液检测方法研究

Study on Detection Method of Gentamicin Fermentation Broth

关键词:  庆大霉素发酵液  检测方法  旋光法  紫外分光光度法  HPLC 法

Key words:  Gentamicin fermentation broth,detection?method?,polarimetry,ultraviolet spectroscopy,HPLC method.

摘要:

目的 在庆大霉素微生物发酵过程中，探索发酵液中庆大霉素效价的快速测定方法。方法 在优化的发酵工艺条件下发酵生产庆大霉素，分别运用旋光法、紫外分光光度法和高效液相色谱法（HPLC）对庆大霉素发酵液的效价进行测定。结果 最佳发酵条件为：种瓶接种量2cm2，种龄24h，发酵瓶接种量12%，发酵周期5d。优化后的庆大霉素发酵液效价相较于优化前提高70%左右。紫外风光光度法、HPLC法和旋光法测定的庆大霉素发酵液效价产生的误差均小于3%。结论 三种检测方法均能运用在庆大霉素发酵液效价的快速检测中。旋光法操作简单但对标准品需求量大，紫外分光光度法耗时短但只能测定庆大霉素总体效价，HPLC法需要出峰时间。因此，在庆大霉素发酵液效价测定中需根据自己的需求选择最佳检测方法。

Abstract:

Objective: To explore the rapid determination method of gentamicin titer in fermentation broth during the microbial fermentation of gentamicin.. Methods: Gentamicin was fermented under optimized fermentation conditions. The potency of gentamicin fermentation broth was determined by optical rotation method, ultraviolet spectrophotometry and high performance liquid chromatography (HPLC). Results: The best fermentation conditions were: seed bottle inoculation volume 2cm2, seed age 24h, fermentation bottle inoculation volume 12%, fermentation period 5d. The optimized potency of gentamicin fermentation broth is increased by about 70% compared with that before optimization. The errors of gentamicin fermentation broth titers determined by ultraviolet wind photometric method, HPLC method and optical rotation method are all less than 3%. Conclusion: The three detection methods can be used in the rapid detection of gentamicin fermentation broth titer. The optical rotation method is simple to operate but has a large demand for standard products. The ultraviolet spectrophotometry method takes a short time but can only determine the overall potency of gentamicin. The HPLC method requires peak time. Therefore, in the determination of gentamicin fermentation broth titer, it is necessary to choose the best detection method according to your needs.