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Rapid and sensitive detection of auxins and flavonoids in plant samples by high‐performance liquid chromatography coupled with tandem mass spectrometry
Author(s) -
Liang Yuan,
Zhu Xiaocui,
Wu Tongbo,
Zhao Meiping,
Liu Huwei
Publication year - 2012
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201200418
Subject(s) - chemistry , chromatography , indole 3 acetic acid , acetic acid , kaempferol , tandem mass spectrometry , mass spectrometry , detection limit , indole test , auxin , quercetin , biochemistry , gene , antioxidant
Simultaneous determination of indole‐3‐acetic acid and methyl indole‐3‐acetic acid ester in small amounts of plant tissue is essential for elucidating their mutual transformation mechanism and the in vivo function of methyl indole‐3‐acetic acid ester. Rapid quantification of flavonoids in the same sample is important for clarifying their roles in the transport of auxins and other phytohormones. Herein, we describe a simple method for the simultaneous determination of indole‐3‐acetic acid and its methyl ester in the roots of the Arabidopsis thaliana seedlings and a protocol for the rapid extraction and quantification of quercetin and kaempferol in these seedlings. High‐performance liquid chromatography coupled with electrospray ionization time‐of‐flight tandem mass spectrometry was used for the detection of all the compounds. Negative data for indole‐3‐acetic acid and positive data for methyl indole‐3‐acetic acid ester were collected in two successive files with a single injection of the extracted sample. Under optimized conditions, the limit of detection for the four compounds was 2 ng/mL for indole‐3‐acetic acid, 0.5 ng/mL for methyl indole‐3‐acetic acid ester, 5 ng/mL for quercetin, and 1 ng/mL for kaempferol, respectively. Because of the high sensitivity of the assay, only 2–10 mg of the plant material was required to obtain quantitative results.