色谱

色谱

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基质固相分散萃取-高效液相色谱-串联质谱法分析拟南芥中的赤霉素

王璐1, 吴倩2, 段春凤2, 吴大朋2, 关亚风1,2*   

  1. 1. 中国科学技术大学化学系, 安徽 合肥 230026; 2. 中国科学院分离分析化学重点实验室 中国科学院大连化学物理研究所仪器分析化学室, 辽宁 大连 116023
  • 收稿日期:2011-07-29 修回日期:2011-08-19 出版日期:2011-09-28 发布日期:2011-10-28
  • 通讯作者: 关亚风,博士生导师,研究员,主要研究方向为微型分离分析.
  • 基金资助:

    国家自然科学重点基金项目(No. 90917020)

Determination of gibberellins in Arabidopsis thaliana by matrix solid-phase dispersion extraction and high performance liquid chromatography-tandem mass spectrometry

WANG Lu1, WU Qian2, DUAN Chunfeng2, WU Dapeng2, GUAN Yafeng1,2*   

  1. 1. Department of Chemistry, University of Science and Technology of China, Hefei 230026, China; 2. CAS Key Laboratory of Separation Science for Analytical Chemistry, Department of Instrumentation and Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
  • Received:2011-07-29 Revised:2011-08-19 Online:2011-09-28 Published:2011-10-28

PDF

570

被引次数

40

摘要： 建立了基质固相分散萃取-高效液相色谱-串联质谱法(MSPD-HPLC-MS/MS)同时测定拟南芥中3种赤霉素GA1, GA3和GA4的分析方法。将拟南芥样品与C18填料混合研磨制成MSPD柱,并采用80%冷甲醇洗脱。采用反相C18色谱柱进行分离,以0.05%甲酸水溶液和乙腈为流动相进行梯度洗脱,采用电喷雾离子源负离子模式(ESI~)电离,多反应监测模式检测。对样品前处理条件、色谱分离条件和质谱检测条件进行了优化,结果表明,在最优条件下,3种赤霉素在10～300 ng/g范围内均呈良好线性关系,相关系数(r2)均大于0.98,检出限在1.1～4.1 ng/g之间。在10～50 ng/g添加水平下,平均回收率范围为54.7%～102.6%,相对标准偏差（RSD,n=3)为3.2%～12.8%。该方法操作简单、灵敏度高、选择性好、回收率高,适合拟南芥中GA1、GA3、GA4含量的测定。

关键词: 赤霉素, 高效液相色谱-串联质谱法, 基质固相分散萃取, 拟南芥

Abstract: A method for the analysis of gibberellin A1 (GA1), gibberellin A3 (GA3) and gibberellin A4 (GA4) in Arabidopsis thaliana by matrix solid-phase dispersion extraction (MSPD) and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed. The solid sample of Arabidopsis thaliana was gently blended with C18 to obtain a homogeneous mixture. This mixture was transferred to an SPE cartridge filled with 0.5 g C18 to form a MSPD column. GA1, GA3 and GA4 were eluted with cold 80% methanol aqueous solution. The target compounds were separated on a C18 column with a gradient elution of 0.05% formic acid aqueous solution and acetonitrile as the mobile phase. The identification and quantification were carried out by using electrospray ionization in negative ion mode (ESI~) with multiple reaction monitoring (MRM). The linear ranges for GA1, GA3 and GA4 were all from 10 to 300 ng/g with correlation coefficients greater than 0.98. The limits of detection were in the range of 1.1~4.1 ng/g. The average recoveries and relative standard deviations were 54.7%~102.6% and 3.2%~12.8% respectively in the spiked range of 10~50 ng/g. The method is simple, sensitive, efficient and accurate. It is suitable for the confirmation and quantitative determination of GA1, GA3 and GA4 in Arabidopsis thaliana.

Key words: Arabidopsis thaliana, high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), gibberellins, matrix solid-phase dispersion extraction