高效液相色谱法分析微生物代谢物埃博霉素B的含量

目的建立检测黏细菌纤维堆囊菌发酵液中代谢物埃博霉素B(EpoB)含量的方法。方法用高效液相色谱法测定EpoB的含量,色谱柱为C18柱,流动相为甲醇-水(60∶40),流速为1.0 mL/min,进样量为3μL,检测波长为247 nm。结果平均回收率为99.55%,RSD为1.94%(n=9),精密度试验表明,日内RSD为0.94%(n=5),日间RSD为1.98%(n=5),EpoB在0.625~10 mg/mL浓度范围内呈线性关系。最低检测限为1.25μg/mL。结论此法可以用于检测代谢产物的含量,具有良好的准确度和精密度。

Quantitative analysis of microbic metabolin EpoB by high performance liquid chromatography

Purpose To develop a HPLC method for quantitative analysis of EpoB in the fermentation broth of Sorangium cellulosum.Methods High performance liquid chromtographic system includes a Kromasil C18 column,eluted with methanol-water(60∶40) at the flow rate 1.0 mL/min.Results The average recovery is 99.55% with RSD 1.94% and the precision in a day(RSD)is 0.94% while 1.98% between days.The detection limit is low(1.25 μg/mL) at 247 nm detection.The calibration linear ranges between 0.625 mg/mL and 10 mg/mL.Conclusion The method is feasible for the quantitative determination of EpoB in fermentation broth with high accuracy and precision.