Determination of Endogenous Brassinosteroids in Plant Tissues Using Solid-phase Extraction with Double Layered Cartridge Followed by High-performance Liquid Chromatography-Tandem Mass Spectrometry
Determination of Endogenous Brassinosteroids in Plant Tissues Using Solid-phase Extraction with Double Layered Cartridge Followed by High-performance Liquid Chromatography-Tandem Mass Spectrometry
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双层柱固相萃取高效液相色谱-串联质谱法测定植物组织中的内源油菜素内酯
DOI：
10.1002/pca.2421
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发表时间：
2013-07-01
影响因子：
3.3
通讯作者：
Feng, Yu-Qi
中科院分区：
文献类型：
--
作者：
Ding, Jun;Mao, Li-Jing;Feng, Yu-Qi
植物甾醇是一类重要的植物激素，对植物的生长发育和一系列生理现象起着重要的作用。为了了解BR的生物合成、降解和代谢途径，需要一种可靠的BR分析方法，并采用有效的样品前处理工艺。目的建立一种快速、有效的植物组织中内源性BR的定量分析方法。方法首先用乙腈提取BRs，然后用无水MgSO 4和NaCl脱水，最后用双层固相萃取柱净化，最后用液-液萃取（LLE）进一步净化。纯化过程主要在GCB/PSA柱上实现。GCB可以消除疏水性化合物，特别是含有体系的化合物，PSA可以消除极性干扰。结果内源性BRs在0.4 ~ 500 ng/mL（0.0124 ~ 15 ng）范围内线性关系良好，相关系数均在0.9957以上。该方法的相对回收率在71.1- 113.1%之间，日内和日间相对标准偏差（RSD）均小于16.3%。利用所提出的方法，植物组织量的需求被最小化到1克鲜重，这是迄今为止报道的最小量，据我们所知。版权所有（c）2013约翰威利父子有限公司
Introduction Brassinosteroids (BRs) are a group of important phytohormones that play vital roles in plant growth, development and a series of physiological phenomena. In order to understand biosynthesis, degradation and metabolic pathways of BRs, a reliable analytical method of BRs with effective sample pre-treatment process is favourable. Objective The development of a quick and effective method for the quantification of endogenous BRs in plant tissue with the aid of double layered solid-phase extraction (SPE) cartridges (graphite carbon black and primary secondary amine silica sorbent: GCB/PSA). Method The method involved an initial extraction of BRs with acetonitrile, a dehydration process with anhydrous MgSO4 and NaCl, a SPE purification process with a double layered cartridge, and a further clean-up step utilising liquid-liquid extraction (LLE). The purification process was mainly realised on the GCB/PSA cartridge. GCB could eliminate hydrophobic compounds, especially those containing a system, and PSA was introduced to remove the polar interferences. Endogenous BRs were quantified by HPLC-ESI-MS/MS. Results Good linearities were obtained in the range of 0.4-500ng/mL (0.0124-15ng), with the correlation coefficients above 0.9957. The relative recoveries of BRs of this method were in the range of 71.1-113.1%, with intra- and interday relative standard deviations (RSDs) less than 16.3%. With the proposed method, the requirement of plant tissue amount was minimised to 1g fresh weight, which is the smallest amount reported so far, to our knowledge. Copyright (c) 2013 John Wiley & Sons, Ltd.