右旋磷霉素手性转化产物检测方法的建立

建立以右旋磷霉素为底物生物转化产生左旋磷霉素的检测方法。通过考察藤黄微球菌与大肠埃希菌对左旋磷霉素的最小抑菌浓度,确定检定菌种类;优化薄层色谱展开剂系统,将薄层色谱与生物显影相结合对转化产物进行定性;利用含量标准曲线,使用生物活性检测法对转化产物进行定量。藤黄微球菌对左旋磷霉素的最小抑菌浓度比大肠埃希菌小100倍,因此作为本实验的检定菌;薄层色谱的最佳展开剂确定为正丁醇/甲醇/水(4∶3∶2,体积比),此时左旋磷霉素在生物检定板上呈现边缘清晰的抑菌圈,且Rf值为0.51;在1.43~5.0 mg/mL范围内,抑菌圈直径与磷霉素浓度的对数值成正比。以藤黄微球菌作为检定菌,不需要对转化产物进行分离纯化和精制,可利用薄层色谱与生物显影相结合的方法对产物进行定性,利用生物活性检测法对产物进行定量,达到简便快速地对转化产物进行定性和定量的目的。

Detection Method for Chiral Transformation Product of Dextral-Fosfomycin

In order to establish a detection method for levo-fosfomycin in bio-transformation liquid when dextral-fosfomycin was employed as a substrate,the MIC values of fosfomycin against Micrococcus luteus(Ml) and E.coli(Ec) were determined,which was used to define the test microorganism.The developing system of TLC combined with bio-visualization was optimized to identify the transformed product,and employed bioactivity assay method to quantify the transformed product.The results showed that MIC value of fosfomyci...