HPLC法测定Streptomyces_sp.发酵液中bafilomycin_A1的含量

目的：建立放线菌Streptomyces sp.发酵液中bafilomycin A1的鉴别及含量测定的方法。方法：采用高效液相色谱-四极杆飞行时间串联质谱（HPLC-Q-TOF-MS）分析技术进行定性鉴别；采用高效液相色谱法（HPLC法）进行含量测定，使用Ultimate LP-C₁₈色谱柱（4.6 mm&#215;250 mm，5 μm），流动相为甲醇-水（85：15），流速1.0 mL&#183;min^-1，检测波长247 nm，柱温40℃。结果：bafilomycin A1质量浓度在18.75~600 μg&#183;mL^-1范围内线性良好，相关系数为1.0（n=6）；平均回收率为101.4%，RSD为1.3%（<2.0%）；方法的精密度RSD为1.2%。3批发酵样品中bafilomycin A1含量分别为183.69、196.29、198.63 μg&#183;mL^-1，均在有效测定范围内。结论：本方法可以用于bafilomycin A1发酵、提炼及纯化过程的含量测定。

Objective:To develop an HPLC method for the determination of bafilomycin A1 in Streptomyces sp.fermentation broth.Methods:HPLC-Q-TOF-MS was used to qualitatively identify bafilomycin A1 in fermentation broth.HPLC was used to quantitatively determine the content of bafilomycin A1 in Streptomyces sp.fermentation broth.The separation was achieved on a Ultimate LP-C₁₈ (4.6 mm&#215;250 mm, 5 μm) column at 40℃ and detected at 247 nm, the mobile phase consisted of methanol and water (85:15) at a flow rate of 1.0 mL&#183;min^-1.Results:The calibration curve was linear over the range of 18.75-600 μg&#183;mL^-1 with the correlation coefficient of 1.0 (n=6).The average recovery of bafilomycin A1 was 101.4% (RSD=1.3%).Bafilomycin A1 contents in three batches of samples were 183.69, 196.29, 198.63 μg&#183;mL^-1 within the effective measurement range, respectively.Conclusion:The method is suitable for the determination of bafilomycin A1 in fermentation broth.

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