Comprehensive chemical derivatization for gas chromatography-mass spectrometry-based multi-targeted profiling of the major phytohormones

被引：117

作者：

Birkemeyer, C ^{[1
]}

Kolasa, A ^{[1
]}

Kopka, J ^{[1
]}

机构：

[1] Max Planck Inst Mol Plant Physiol, Dept Willmitzer, D-14476 Golm, Germany

来源：

JOURNAL OF CHROMATOGRAPHY A | 2003年 / 993卷 / 1-2期

关键词：

derivatization; GC; Nicotiana tabacum; Arabidopsis thaliana; tobacco; organic acids; plant hormones; auxin; cytotoxin;

D O I：

10.1016/S0021-9673(03)00356-X

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

In the present investigation we report selection of the N-methyl-N-(tert.-butyldimethylsilyl)trifluoroacetamide (MTBSTFA) reagent as the most comprehensive derivatization protocol among 17 tested reactions covering trifluoroacetylation, pentafluorobenzylation, methylations, and trimethylsilylations. MTBSTFA allowed easy and robust tert.-butyldimethylsilyl derivatization of 1-aminocyclopropane-1-carboxylic acid, indole-3-acetic acid, (+/-)-jasmonic acid, salicylic acid, (+/-)-abscisic acid, meta-topolin, and trans-zeatin. Detection limits as analysed by selected ion monitoring quadrupole GC-MS were 0.2, 0.01, 1.0, 0.02, 0.3, 0.3, and 0.9 pmol of injected substance, respectively. Analysis of gibberellic acid A3, trans-zeatin riboside and (+/-)-abscisic acid-beta-D-glucopyranosyl ester was best when coupled by splitting extracts and trimethysilylation. The MTBSTFA derivatization protocol was optimised, and validated. The preparation was insensitive to 2% residual water and to :51 day storage at room temperature. The final scheme was highly reproducible and successfully applied to extracts from similar to300 mg (fresh mass) of tobacco (Nicotiana tabacum) root and Arabidopsis thaliana seedling. (C) 2003 Elsevier Science B.V. All rights reserved.

引用

页码：89 / 102

页数：14

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