| Metabolite Profiling of Mature Arabidopsis thaliana Seeds Using Gas Chromatography-Mass Spectrometry (GC-MS)
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Author:
Date:
2016-11-05
[Abstract] Metabolite profiling using gas chromatography-mass spectrometry (GC-MS) permits the annotation and quantification of a relatively wide variety of metabolites, covering a wide range of biochemical groups of metabolites. Lisec et al. (2006) established a method for GC-MS profiling in plants. Based on this protocol, we provide here a detailed GC-MS-based metabolite profiling protocol to identify compounds belonging to several biochemical groups in the primary metabolism of mature Arabidopsis thaliana seeds (Cohen et al., 2014). The protocol uses methoxyamine hydrochloride and N-methyl-N-trimethylsilyltriflouroacetamide (MSTFA) as derivatization reagents, as previous studies indicated these are the most appropriate compounds for profiling of plant ...
[摘要] 使用气相色谱 - 质谱（GC-MS）的代谢物分析允许注释和量化相对多种代谢物，涵盖广泛的代谢物的生物化学组。 Lisec等人。 （2006）建立了一种在植物中进行GC-MS分析的方法。基于该方案，我们在这里提供详细的基于GC-MS的代谢物分析方案以鉴定属于成熟拟南芥种子的主要代谢中的几个生物化学基团的化合物（Cohen等人。，2014）。该方案使用甲氧基胺盐酸盐和N，N-甲基-N，N-三甲基甲硅烷基三氟乙酰胺（MSTFA）作为衍生化试剂，如先前的研究所表明的，这些是用于分析植物代谢物的最合适的化合物。该方案相对快速，提供可重复的结果，并且可以用于简化许多其它类型的植物组织的代谢物，只有很少的修改。在这种情况下，开发种子可以作为研究代谢调节的优秀系统，因为在它们的开发期间，储备化合物的大规模合成在严格的转录调节下发生控制并且与时间上不同的代谢开关相关。
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| Analysis of Flower Cuticular Waxes and Cutin Monomers
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Author:
Date:
2013-09-20
[Abstract] Here we describe procedures for the flower cuticular waxes extraction, modification and subsequent qualitative and quantitative analysis by gas-chromotography-mass spectrometry (GC-MS) and gas-chromotography with flame ionization detector (GC-FID), accordingly. To characterize flower cutin monomers two experimental setup are described: (i) analysis of enzymatically isolated cuticles in order to determine the relative proportions of cutin monomers; (ii) analysis of freeze-dried material for quantitative estimation of the cutin content. This report is an adaptation of the earlier published protocols developed for the chemical analysis of the cuticles in vegetative organs (Leide et al., 2007).
[摘要] 在这里我们介绍花角质蜡提取，修改和随后的气相色谱 - 质谱（GC-MS）和气相色谱与火焰离子化检测器（GC-FID）的定性和定量分析的程序。 为了表征花角质单体，描述了两个实验装置：（i）分析酶分离的角质层，以确定角质单体的相对比例; （ii）冷冻干燥材料的分析，用于定量估计角质含量。 该报告是对早期公开的用于营养器官中角质层的化学分析的协议的改编（Leide等人，2007）。
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