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反相高效液相色谱法测定发酵液中FR-008/杀念菌素及其相关组分的含量

毛相朝, 沈亚领*, 魏东芝**, 陈实, 邓子新   

  1. 1.华东理工大学生物反应器国家重点实验室鲁华生物技术研究所, 上海 200237;
    2.上海交通大学生命科学与技术学院BIOX生命科学研究中心, 上海 200030
  • 收稿日期:2004-09-18 修回日期:2005-05-10 出版日期:2005-06-15 发布日期:2005-06-15
  • 通讯作者: 沈亚领*, 魏东芝**

Determination of Candicidin/FR-008 and Related Components in Fermentation Broth by RP-HPLC

MAO Xiang-zhao, SHEN Ya-ling*, WEI Dong-zhi**, CHEN Shi, Deng Zi-xin   

  1. 1.State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai 200237, China;
    2.Bio-X Life Science Research Center and School of Life Science and Biotechnology, Shanghai Jiaotong University, Shanghai 200030, China
  • Received:2004-09-18 Revised:2005-05-10 Online:2005-06-15 Published:2005-06-15
  • Contact: SHEN Ya-ling*, WEI Dong-zhi**

摘要: 目的 建立发酵液中杀念菌素(Candicidin)/FR008及其相关组分的快速测定方法。方法 色谱条件SB-C8(4.6 mm×250 mm)色谱柱, 乙腈-20 mmol·L-1醋酸铵缓冲液(pH4.0) = 40:60为流动相, 流速为1.0 mL·min-1, 柱温为25 ºC, 检测波长380nm结果 6.25-500 μg·mL-1范围内Candicidin/FR008与峰面积呈良好线性关系, 回归方程为Y = 20.461x + 30.748, r = 0.9991。精密度和重复性分别为1.84%3.8%结论 本方法的精密度、回收率和线性关系均表现较好, 且具有精确、高效、操作简单等特点, 适用于发酵产物的常规检测。

关键词: 杀念菌素/FR-008, 杀念菌素/FR-008, 杀念菌素/FR-008, 高效液相色谱, 高效液相色谱, 高效液相色谱, 发酵液, 发酵液, 发酵液, 衍生物, 衍生物, 衍生物

Abstract: Aim A liquid chromatographic method for the determination of candicidin/FR-008 and related components in fermentation broth has been developed. Methods There were four major components in the candicidin/FR-008 complex, which were separated by HPLC under the following conditions: SB-C8 column (4.6 mm ×250 mm, 5 μm) was used, the mobile phase consisted of acetonitrileammonium acteate (20 mmol·L-1, pH 4.0) (40:60, V/V), with a flow rate of 1.0 mL·min-1 the UV detection wavelength was 380 nm, and the whole process was performed at 25 ºC. Results The linearity was obtained in the range of 6.25-500 μg·mL-1 candicidin/FR-008 with the regression equation of Y = 20 461x + 30 748 and the correlation coefficient of 0.999 1. The instrument precision was 1.84% and the method precision was 3.8%. Conclusion This method is accurate, rapid and simple; it can be used for determination of candicidin/FR-008 and related components in fermentation broth.

Key words: candicidin/FR-008, candicidin/FR-008, HPLC, HPLC, fermentation broth, fermentation broth, metabolites, metabolites

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Supporting: *Corresponding author. Tel.: 86-21-64253156, 86-21-64252078