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发酵法生产紫杉醇的检测分析
引用本文:金涛,周东坡,孙宇石,刘利群,邹积宏,平文祥,阎秀峰.发酵法生产紫杉醇的检测分析[J].生物技术,2012,22(3):65-69.
作者姓名:金涛  周东坡  孙宇石  刘利群  邹积宏  平文祥  阎秀峰
作者单位:1. 黑龙江大学生命科学学院、微生物黑龙江省高校重点实验室,黑龙江哈尔滨150080;农业微生物技术教育部工程研究中心博士后科研工作站,黑龙江哈尔滨150500;东北林业大学博士后科研流动站,黑龙江哈尔滨150086
2. 黑龙江大学生命科学学院、微生物黑龙江省高校重点实验室,黑龙江哈尔滨,150080
3. 黑龙江省食品药品检验检测所,黑龙江哈尔滨,150001
4. 东北林业大学博士后科研流动站,黑龙江哈尔滨,150086
基金项目:黑龙江省教育厅科学技术研究项目,中国博士后科学基金会,黑龙江大学高层次人才(创新团队)支持计划
摘    要: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。

关 键 词:检测  发酵  紫杉醇

Detection of Taxol Produced by Fermentation
JIN Tao , ZHOU Dong-po , SUN Yu-shi , LIU Li-qun , ZOU Ji-hong , PING Wen-xiang , YAN Xiu-feng.Detection of Taxol Produced by Fermentation[J].Biotechnology,2012,22(3):65-69.
Authors:JIN Tao  ZHOU Dong-po  SUN Yu-shi  LIU Li-qun  ZOU Ji-hong  PING Wen-xiang  YAN Xiu-feng
Institution:1.Laboratory of Microbiology,College of Life Science,Heilongjiang University,Harbin 150080; 2.Postdoctoral Research Working Station of Engineering Research Center of Agricultural Microbiology Technology,Ministry of Education,Harbin 150500; 3.Postdoctoral Research Station,Northeast Forestry University,Harbin 150086; 4.Heilongjiang Provincial Institute For Food&Drug Control,Harbin 150001,China)
Abstract:Objective:The content of taxol in fermentation broth was detected by HPLC.Method:The analytic samples were obtained from ethyl acetate extract of fermentation product of taxol producing strain.The HPLC separation was performed with benzene analytical column(250mm×4.6mm,5μm)in the flow rate of 1mL/min,the mobile phase was acetonitrile-methol-water(36∶ 4∶ 60),the injection volume was 20mL,the column temperature was room temperature.Result:Taxol and interfering substances can reach baseline separation.The calibration curve of peak area and taxol concentration was linear in the concentration range of 2.2-110μg/mL with good correlation coefficient(r=0.999 6).The average recovery rate was 99.55%,RSD 0.60%.Conclusion:The convenient analytical method on benzene column with high sensitivity and high accuracy did not require complicated sample pretreatment and complex instrument configuration,could be used to determine the content of taxol in fermentation broth effectively,compared to chromatographic condition on C18 column.
Keywords:detection  fermentation  taxol
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