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目的:采用高效液相色谱法对发酵液中的紫杉醇进行测定。方法:将紫杉醇产生菌发酵产物经乙酸乙酯萃取得测定样品,高效液相色谱分析方法为苯基柱(250mm×4.6mm,5μm),流动相乙腈-甲醇-水(36∶4∶60),流速1mL/min,检测波长227nm,进样体积20μL,柱温室温。结果:紫杉醇与干扰物可达到基线分离,在2.2～110μg/mL范围内,紫杉醇的峰面积与浓度线性关系良好,相关系数0.9996,平均回收率为99.55%,RSD为0.60%。结论:与使用C18柱色谱条件相比,该分析方法灵敏度高,不需要复杂的样品前处理过程,仪器配置不复杂、操作方便、准确性高,可有效地检测发酵液中紫杉醇的含量。
OBJECTIVE: To determine paclitaxel in fermentation broth by high performance liquid chromatography. Methods: The product was extracted with ethyl acetate. The high performance liquid chromatographic method was phenyl column (250mm × 4.6mm, 5μm). The mobile phase was acetonitrile - methanol - water (36:4:60) , Flow rate 1mL / min, detection wavelength 227nm, injection volume 20μL, column temperature room temperature. Results: Paclitaxel and interferon could be separated baseline. Within the range of 2.2-110 μg / mL, the peak area of paclitaxel was linear with the concentration. The correlation coefficient was 0.9996, the average recovery was 99.55% and the RSD was 0.60%. Conclusion: Compared with C18 column chromatography, this method has high sensitivity and does not require complex sample preparation. The instrument configuration is not complicated, easy to operate and high accuracy, which can effectively detect the content of paclitaxel in the fermentation broth.