Analysis of low abundant trehalose-6-phosphate and related metabolites in Medicago truncatula by hydrophilic interaction liquid chromatography–triple quadrupole mass spectrometry
| Autor: | João Ferreira, Diana Branco, Maria Rosário Bronze, Susana de Sousa Araújo, Pedro Fevereiro, Carla António, Tiago F. Jorge, Ana Teresa Mata |
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| Rok vydání: | 2016 |
| Předmět: |
0106 biological sciences
0301 basic medicine Sucrose Metabolite Tandem mass spectrometry 01 natural sciences Biochemistry Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound Limit of Detection Tandem Mass Spectrometry Medicago truncatula Methylphosphonic acid Chromatography biology Hydrophilic interaction chromatography Organic Chemistry Selected reaction monitoring Trehalose General Medicine Reference Standards biology.organism_classification Biosynthetic Pathways Triple quadrupole mass spectrometer Plant Leaves Solutions 030104 developmental biology chemistry Metabolome Sugar Phosphates Hydrophobic and Hydrophilic Interactions Chromatography Liquid 010606 plant biology & botany |
| Zdroj: | Journal of Chromatography A. 1477:30-38 |
| ISSN: | 0021-9673 |
| DOI: | 10.1016/j.chroma.2016.11.031 |
| Popis: | Trehalose-6-phosphate (T6P) is an important signaling metabolite involved in plant growth control that inhibits the sucrose nonfermenting-1-related protein kinase 1 (SnRK1), a key regulator of energy and carbon metabolism in plants. The quantification of T6P in plant tissues is fundamental to improve our understanding of sugar signaling and the links between plant growth and development in response to stress conditions. However, the almost undetectable levels of T6P together with the complex plant matrix and the presence of T6P isomers such as sucrose-6-phosphate (S6P), makes the detection of this metabolite challenging. This work describes the development and validation of a hydrophilic interaction chromatography (HILIC) method for the on-line coupling with negative ion electrospray (ESI) triple quadrupole tandem mass spectrometry (MS/MS) in the highly sensitive and selective multiple reaction monitoring (MRM) mode for the target analysis of metabolic intermediates of the biosynthesis of trehalose, including glucose-6-phosphate (G6P), uridine 5-diphospho-glucose (UDPG), T6P (and its isomer S6P). Enhanced signal in the MRM mode and improved chromatographic separation for each compound were obtained using piperidine and methylphosphonic acid as additives in the HILIC mobile phase. The optimized HILIC-ESI-QqQ-MS/MS method increases the range of sensitive analytical methodologies for the quantification of key low-abundant metabolites, and was applied to quantify the fluctuations of S6P, T6P and G6P in Medicago truncatula plants in response to environmental stress. The levels of S6P, T6P, and G6P in M. truncatula plant tissues (roots and leaves) exposed to a water deficit and recovery treatment, ranged from 30 to 150pmolg-1 FW, 16-120pmolg-1 FW, and 330-1690pmolg-1 FW, respectively. |
| Databáze: | OpenAIRE |
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