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首页> 中文期刊> 《实验室研究与探索》 >UPLC-ESI-MS/MS法测定发酵液中紫杉醇含量

UPLC-ESI-MS/MS法测定发酵液中紫杉醇含量

         
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摘要

产紫杉醇菌发酵液经乙酸乙酯萃取后,应用Waters ACQUITY UPLC仪和Waters Quattro Premier三级四极杆质谱仪检测建立了产紫杉醇菌发酵液中紫杉醇含量的快速测定方法.采用Waters zXTerra MS C18色谱柱(2.1 mm×150 mm,5μm),梯度洗脱,流速0.3 mL/min,柱温为30 ℃;进样量3.0 μL 以超高效液相色谱分离、电喷雾离子化串联质谱分析在多反应监测(MRM)模式下进行检测 在10～100 μg/mL,紫杉醇对照品的峰面积与浓度线性关系良好,相关系数0.998;方法具有良好的精密度,RSD为0.51％;稳定性试验表明室温下样品在8h内稳定;具有良好的重现性,RSD为1.6％;平均加样回收率为99.58％,RSD为0.62％ 采用MRM模式测定紫杉醇含量不仅可以增加定量准确性,而且专一性强,干扰因素少,还为产紫杉醇微生物的快速筛选提供科学的方法.%Waters ACQUITY UPLC and Waters Qualtro Premier third grade quadrupole mass spectrometer were applied to establish a rapid and accurate determination method for the content of taxol in fermentation broth of taxol-yielding fungus by UPLC-ESI-MS/MS after the extraction of fermentation broth by ethyl acetate. The separation was performed with Waters zXTerra MS C18 analytical column(2. 1 mm × 150 mm, 5 μm)with a gradient elution in the flow rate of 0. 3 mL/min. The injection volume was 3.0 (xL, and the column temperature was adjusted to 30 ℃. The detection was performed by the separation of UPLC coupled with the analysis of ES1-MS-MS in the multiple reaction monitoring ( MRM) mode. The calibration curve of peak area and taxol concentration was linear in the concentration range of 10 to rn100 μg/mL with good correlation coefficient (r =0.998). The validation data were indicative of good precision ( RSD 0.51% ). Stability study showed that the samples were stable at room temperature within 8 hours. Good reproducibility (RSD 1.6% ) was also obtained. The average recovery rate was 99.58% , RSD 0.62%. An analytical method for the determination of taxol via UPLC-ESI-MS/MS in MRM could not only increase the quantitative accuracy and possess strong specificity and less interference, but also provide a rapid scientific method capably applied to the screening of taxol-producing microbes.

著录项

  • 来源
    《实验室研究与探索》 |2012年第11期|15-18|共4页
  • 作者

    金涛; 刘盛春; 周东坡; 平文祥; 宋福强; 阎秀峰;

  • 作者单位

    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;

    黑龙江大学物理科学与技术学院,黑龙江哈尔滨150080;

    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;

    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;

    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;

    东北林业大学博士后科研流动站,黑龙江哈尔滨150086;

  • 原文格式 PDF
  • 正文语种 chi
  • 中图分类 药物含量测定;
  • 关键词

    UPLC-ESI-MS/MS; 发酵; 紫杉醇;

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