HPLC法测定Streptomyces_sp.BOR-0331发酵液中疏螺旋体素的含量

目的：建立放线菌Streptomyces sp.BOR-0331发酵液中疏螺旋体素含量测定的方法。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；高效液相色谱（HPLC）法进行含量测定，采用Agilent ZORBAX SB-C₁₈色谱柱（5 μm，4.6 mm×250 mm），流动相为乙腈-0.1%三氟乙酸溶液（50：50），流速为1.5 mL·min^-1，检测波长为257 nm。结果：疏螺旋体素浓度范围在12.5~200 μg·mL^-1线性良好，相关系数为0.9997（n=5）；平均回收率为102.8%（RSD为0.86%），精密度RSD为1.3%；3批发酵液中疏螺旋体素的含量分别为61.18、69.92、61.08 μg·mL^-1。结论：方法准确可靠，可以用于疏螺旋体素发酵、提炼及纯化过程的含量测定。

Objective: To develop an HPLC method for the determination of borrelidinin in fermentation broth of Streptomyces sp. BOR-0331. Methods: HPLC-Q-TOF-MS was used to qualitatively determine borrelidin in fermentation broth,and HPLC was used to quantitatively determine the content of borrelidin. Sample was achieved on a ZORBAX SB-C₁₈ (5 μm, 4.6 mm×250 mm) column and detected at 257 nm. The mobile phase consisted of acetonitrile and 0.1% trifluoroacetic acid solution (50:50,v/v) at a flow rate of 1.5 mL·min^-1. Results: The calibration curve was linear over the range of 12.5-200 μg·mL^-1 with the correlation of 0.999 7 (n=5). The average recovery of borrelidin was 102.8% (RSD=0.86%). The contents of borrelidinin in three batches of fermentation broths were 61.18,69.92 and 61.08 μg·mL^-1,respectively. Conclusion: The method is accurate,simple with good reproducibility,which is suitable for the determination of borrelidinin in fermentation broth.

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