白刺链霉菌(Streptomyces albospinus)CT205产环己酰亚胺含量测定方法的建立

Determination of Cycloheximide Content Produced by Streptomyces albospinus CT205

建立白刺链霉菌(Streptomyces albospinus)CT205代谢产环己酰亚胺含量的检测方法,为菌株CT205的开发利用提供技术支持。采用高效液相色谱法(HPLC)、生物活性法和分光光度法等三种检测手段分别测定菌株CT205发酵上清液及粗提物中活性物质环己酰亚胺(Cycloheximide)的含量。三种方法测定环己酰亚胺标准品均具有较好的线性关系,其中相关系数R_(HPLC)^(2)法(0.9992)>R_(生物活性法)^(2)(0.9937)≈R_(分光光度法)^(2)(0.9965)。HPLC法及生物活性法分别测定CT205发酵液及粗提物中活性物质含量分别为72.87、1555.70及66.15、1259.00μg/mL,HPLC法与生物活性法测定的发酵上清液中活性物质含量误差在+6.72μg/mL。HPLC检测方法的准确性及灵敏度均高于生物活性法,适用于样品含量的准确测定,生物活性法适用于菌株诱变筛选及条件优化实验产生的大批量样品的测定及比较,分光光度法不适用检测杂质含量较多的样品。

The purpose of this study was to establish a method for detecting the content of Cycloheximide produced by Streptomyces albospinus CT205,and to provide technical support for the development and utilization of strain CT205.The content of Cycloheximide in the fermented supernatant and crude extract from strain CT205 were determined with three detection methods including high performance liquid chromatography(HPLC),biological activity and spectrophotometry.The results showed that three methods for determining Cycloheximide standard had a great linear relationship,in which the correlation coefficient R_(HPLC)^(2)(0.9992)>R_(biological activity)^(2)(0.9937)≈R_(spectrophotometry)^(2)(0.9965).The content of active substances in CT205 fermented broth and crude extracts were determined with HPLC and biological activity methods were 72.87,1557.50 and 66.15,1259.00μg/mL respectively.It was found that the error of the active substance content in the fermentation supernatant by HPLC method and biological activity method was+6.72μg/mL.Therefore,HPLC method was more accurate and sensitive than the biological activity method,and was suitable for accurate determination of sample content.The biological activity method was suitable for the determination and comparison of large quantities of samples produced by strain mutagenesis screening and condition optimization experiments,and the spectrophotometry method was not suitable for detecting samples with a large amount of impurities.