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The determination of 22 natural brassinosteroids in a minute sample of plant tissue by UHPLC-ESI-MS/MS
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SYSNO ASEP 0463987 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title The determination of 22 natural brassinosteroids in a minute sample of plant tissue by UHPLC-ESI-MS/MS Author(s) Tarkowská, Danuše (UEB-Q) RID, ORCID
Novák, Ondřej (UEB-Q) RID, ORCID, SAI
Oklešťková, Jana (UEB-Q) RID, ORCID, SAI
Strnad, Miroslav (UEB-Q) RID, ORCIDSource Title Analytical and Bioanalytical Chemistry. - : Springer - ISSN 1618-2642
Roč. 408, č. 24 (2016), s. 6799-6812Number of pages 14 s. Language eng - English Country DE - Germany Keywords Brassinosteroids ; Solid-phase extraction ; Ultra-high performance liquid chromatography Subject RIV EB - Genetics ; Molecular Biology R&D Projects LO1204 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) LK21306 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) GA14-34792S GA ČR - Czech Science Foundation (CSF) Institutional support UEB-Q - RVO:61389030 UT WOS 000382883300027 DOI 10.1007/s00216-016-9807-2 Annotation The triterpenoid plant hormones brassinosteroids (BRs) are believed to influence almost every aspect of plant growth and development. We have developed a sensitive mass spectrometry-based method for the simultaneous profiling of twenty-two naturally occurring brassinosteroids including biosynthetic precursors and the majority of biologically active metabolites. Using ultra-high performance liquid chromatographic (UHPLC) analysis, the run time was reduced up to three times (to 9 min) in comparison to standard HPLC BRs analyses, the retention time stability was improved to 0.1-0.2 % RSD and the injection accuracy was increased to 1.1-4.9 % RSD. The procedures for extraction and for two-step purification based on solid-phase extraction (SPE) were optimised in combination with subsequent UHPLC analysis coupled to electrospray ionisation tandem mass spectrometry (ESI-MS/MS) using Brassica flowers and Arabidopsis plant tissue extracts. In multiple reaction monitoring (MRM) mode, the average detection limit for BRs analysed was close to 7 pg, and the linear range covered up to 3 orders of magnitude. The low detection limits for this broad range of BR metabolites enabled as little as 50 mg of plant tissue to be used for quantitative analyses. The results of determinations exploiting internal standards showed that this approach provides a high level of practicality, reproducibility and recovery. The method we have established will enable researchers to gain a better understanding of the dynamics of the biosynthesis and metabolism of brassinosteroids and their modes of action in plant growth and development. Workplace Institute of Experimental Botany Contact David Klier, knihovna@ueb.cas.cz, Tel.: 220 390 469 Year of Publishing 2017
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