An UPLC-MS/MS method for highly sensitive high-throughput analysis of phytohormones in plant tissues
| Autor: | Andrej Frolov, Bettina Hause, Gerd Ulrich Balcke, Vinzenz Handrick, Anja Henning, Alain Tissier, Mandy Fichtner, Nick Bergau, Hagen Stellmach |
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| Jazyk: | angličtina |
| Předmět: |
Analyte
Jasmonates Solid phase extraction (SPE) Plant stress response Plant Science lcsh:Plant culture Biology Tandem mass spectrometry Electrospray ionization Quantification Botany Genetics lcsh:SB1-1110 Sample preparation Solid phase extraction LC-MS/MS lcsh:QH301-705.5 Organism Chromatography Abiotic stress Plant Part Methodology Highly sensitive Phytohormones lcsh:Biology (General) Wounding Biotechnology |
| Zdroj: | Plant Methods Plant Methods, Vol 8, Iss 1, p 47 (2012) |
| ISSN: | 1746-4811 |
| DOI: | 10.1186/1746-4811-8-47 |
| Popis: | Background Phytohormones are the key metabolites participating in the regulation of multiple functions of plant organism. Among them, jasmonates, as well as abscisic and salicylic acids are responsible for triggering and modulating plant reactions targeted against pathogens and herbivores, as well as resistance to abiotic stress (drought, UV-irradiation and mechanical wounding). These factors induce dramatic changes in phytohormone biosynthesis and transport leading to rapid local and systemic stress responses. Understanding of underlying mechanisms is of principle interest for scientists working in various areas of plant biology. However, highly sensitive, precise and high-throughput methods for quantification of these phytohormones in small samples of plant tissues are still missing. Results Here we present an LC-MS/MS method for fast and highly sensitive determination of jasmonates, abscisic and salicylic acids. A single-step sample preparation procedure based on mixed-mode solid phase extraction was efficiently combined with essential improvements in mobile phase composition yielding higher efficiency of chromatographic separation and MS-sensitivity. This strategy resulted in dramatic increase in overall sensitivity, allowing successful determination of phytohormones in small (less than 50 mg of fresh weight) tissue samples. The method was completely validated in terms of analyte recovery, sensitivity, linearity and precision. Additionally, it was cross-validated with a well-established GC-MS-based procedure and its applicability to a variety of plant species and organs was verified. Conclusion The method can be applied for the analyses of target phytohormones in small tissue samples obtained from any plant species and/or plant part relying on any commercially available (even less sensitive) tandem mass spectrometry instrumentation. |
| Databáze: | OpenAIRE |
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