全自动在线固相萃取-液相色谱-串联质谱法同时检测水稻中6种内源性植物激素  

An Automated On-line Solid Phase Extraction-Liquid Chromatography-Tandem Mass Spectrometric Method for Six Endogenous Phytohormones Analysis in Rice

作　　者：夏群[1,2] 辛培勇[1] 褚金芳[1] 

机构地区：[1]中国科学院遗传与发育生物学研究所国家植物基因研究中心(北京),北京100101 [2]中国科学院大学,北京100049

出　　处：《分析化学》2016年第3期409-415,共7页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.91417314);中科院仪器设备功能开发技术创新项目和中科院关键技术人才项目资助~~

摘　　要：建立了同时检测水稻中6种内源性植物激素脱落酸(Abscisic acid,ABA)、吲哚-3-乙酸(Indole-3-acetic acid,IAA)、水杨酸(Salicylic acid,SA)、茉莉酸(Jasmonic acid,JA)、吲哚-3-丙酸(Indole-3-propionic acid,IPA)和吲哚-3-丁酸(Indole-3-butyric acid,IBA)的全自动在线固相萃取-液相色谱-串联质谱方法。植物样品经过甲醇提取,采用C_(18)固相萃取柱富集净化,流动相将待测物洗脱至C_(18)分析色谱柱进行分离,最终使用串联四极杆质谱进行检测。方法的线性范围为8~320μg/L,相关系数为R^2≥0.99;方法的检出限(S/N=3)范围为0.1~0.8μg/kg;实际样品中方法回收率范围为71.2%~126%,RSD<13%。应用本方法快速、准确地检测了水稻幼穗中多种内源性植物激素的含量,并与目前植物学领域内常用的检测方法进行了比较。同时,本方法对水稻受伤叶片的内源植物激素含量变化进行了定量分析,其含量随受伤时间的变化趋势与其生物背景的实验结果相吻合。An automated on-line solid phase extraction-liquid chromatography-tandem mass spectrometry（ on-line SPE-LC-MS /MS） method for the simultaneous quantification of six endogenous phytohormones including abscisic acid（ ABA）,indole-3-acetic acid（ IAA）,salicylic acid（ SA）,jasmonic acid（ JA）,indole-3-propionic acid（ IPA） and indole-3-butyric acid（ IBA） in rice tissues was described. Plant samples were extracted with methanol and on-line SPE procedure was performed with C（18） SPE cartridges. Analytes were eluted to C（18） analytical column with mobile phase and further analyzed by LC-MS / MS. The performance of the method was fully validated. Linearity showed good correlation（ R^2≥0. 99）. Limits of detections（ S /N = 3）were in the range of 0. 1-0. 8 μg / kg. Recoveries varied between 71. 2% and 126%. The method was rapid and sensitive to determine multiple phytohormones in rice young panicles and the results were cross-validated with the off-line SPE-LC-MS / MS phytohormone analytical method. Finally,the method was applied to monitor the changes of ABA,IAA,SA and JA in wounded rice leaves. The trends were in accord with plant physiological processes.

关 键 词：植物激素 在线固相萃取-液相色谱-串联质谱 定量分析 水稻 

分 类 号：S511[农业科学—作物学] O657.63[理学—分析化学]