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Quantitative analysis of major plant hormones in crude plant extracts by high-performance liquid chromatography–mass spectrometry

Nature Protocols volume 5, pages 986–992 (2010)Cite this article

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Abstract

The ability to measure plant hormones quantitatively is important as plant hormones regulate plant growth, development and response to biotic and abiotic cues. In this protocol, we describe the quantitative analysis of major plant hormones from crude plant extracts. Plant hormones are determined using reverse-phase liquid chromatography–tandem mass spectrometry with multiple reaction monitoring. The method provides quantification of most major plant hormones in a single run from 50 mg of fresh plant tissue. Extraction and quantitative analysis of 40 samples takes 2–3 d.

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Figure 1: Structures of endogenous plant hormones.
Figure 2: Schematic flow chart showing the steps for the analysis of 18 endogenous plant hormones and their metabolites in plant tissues by HPLC–electrospray ionization–tandem mass spectrometry (HPLC–ESI–MS/MS).
Figure 3: Typical MS/MS spectra of authentic standards and MRM chromatograms of detected plant hormones in 50 mg of fresh Arabidopsis leaf crude extracts.
Figure 4: Amounts of plant hormones and their metabolites in fresh Arabidopsis leaves, determined by LC–ESI–MS/MS.

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Acknowledgements

This work was supported by the National Science Foundation (MCB-0455318, DBI-0521250 and IOS-0818740) and a grant from the US Department of Agriculture (2007-35318-18393).

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Author notes

  1. Xiangqing Pan

    Present address: Current address: Solae, LLC, 4300 Duncan Avenue, St. Louis, MO 63110, USA.,

Authors and Affiliations

  1. Department of Biology, University of Missouri, St. Louis, MO, USA

    Xiangqing Pan & Xuemin Wang

  2. Donald Danforth Plant Science Center, St. Louis, MO, USA

    Xiangqing Pan & Xuemin Wang

  3. Division of Biology, Kansas Lipidomics Research Center, Kansas State University, Manhattan, KS, USA

    Ruth Welti

Authors
  1. Xiangqing Pan
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Contributions

X.P. contributed to the development and evaluation of the methods and combined the data and wrote the protocol. R.W. and X.W. supervised and drove the projects to use LC–MS/MS for plant hormone analysis and contributed to writing and improving the protocols. This work was done in X.W.'s lab based at Donald Danforth Plant Science Center.

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Correspondence to Xiangqing Pan.

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The authors declare no competing financial interests.

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Pan, X., Welti, R. & Wang, X. Quantitative analysis of major plant hormones in crude plant extracts by high-performance liquid chromatography–mass spectrometry. Nat Protoc 5, 986–992 (2010). https://doi.org/10.1038/nprot.2010.37

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