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Trace analysis of multi-class phytohormones in Oryza sativa using different scan modes in high-resolution Orbitrap mass spectrometry : method validation, concentration levels, and screening in multiple accessions

Ashley Haeck (UGent) , Herman Van Langenhove (UGent) , Lies Harinck (UGent) , Tina Kyndt (UGent) , Godelieve Gheysen (UGent) , Monica Höfte (UGent) and Kristof Demeestere (UGent)
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Abstract
Phytohormones are signaling and regulating metabolites involved in numerous plant processes, including growth, development, and responses to stress. Currently, the focus is on the analysis of multiple phytohormones in order to characterize crosstalk and hormone signaling networks. In this paper, representative phytohormones of the major classes are simultaneously determined in rice tissues by a generic solid-liquid extraction, followed by liquid chromatography and electrospray ionization high-resolution tandem mass spectrometry using a Q-Exactive (TM) instrument. After a thorough optimization of the sample preparation, the analytical method was fully validated toward the ultra-trace quantification of six a priori selected plant hormones using three scan modes of the quadrupole-Orbitrap instrument: full-scan high-resolution mass spectrometry, targeted single ion monitoring (t-SIM), and t-SIM followed by data-dependent tandem mass spectrometry. Overall, a similar quantitative performance was noticed for the different scan modes. The analytical method was successfully applied to measure basal phytohormone levels in six different rice accessions, comprising Oryza sativa ssp. japonica, indica, and Oryza glaberrima. Hormone concentrations were higher in shoots than in roots or at least similar. Except for a lower level of salicylic acid in shoots of O. glaberrima versus O. sativa, no other differences in hormone levels could be noticed that were dependent of the (sub)species assignment of the analyzed accessions. Making use of the benefits of full-scan high-resolution mass spectrometry, a first post-run suspect screening was performed, suggesting - based on accurate mass measurements and isotopic patterns - the possible presence of about 50 additional plant hormones in the rice tissues.
Keywords
PLANT HORMONES, ABSCISIC-ACID, SALICYLIC-ACID, QUANTITATIVE-ANALYSIS, DETECTION LIMITS, JASMONIC ACID, UPLC-MS/MS, RICE, QUANTIFICATION, EXTRACTION, Plant metabolites, Phytohormones, High-resolution mass spectrometry, Orbitrap, Rice, Accessions

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Chicago
Haeck, Ashley, Herman Van Langenhove, Lies Harinck, Tina Kyndt, Godelieve Gheysen, Monica Höfte, and Kristof Demeestere. 2018. “Trace Analysis of Multi-class Phytohormones in Oryza Sativa Using Different Scan Modes in High-resolution Orbitrap Mass Spectrometry : Method Validation, Concentration Levels, and Screening in Multiple Accessions.” Analytical and Bioanalytical Chemistry 410 (18): 4527–4539.
APA
Haeck, A., Van Langenhove, H., Harinck, L., Kyndt, T., Gheysen, G., Höfte, M., & Demeestere, K. (2018). Trace analysis of multi-class phytohormones in Oryza sativa using different scan modes in high-resolution Orbitrap mass spectrometry : method validation, concentration levels, and screening in multiple accessions. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 410(18), 4527–4539.
Vancouver
1.
Haeck A, Van Langenhove H, Harinck L, Kyndt T, Gheysen G, Höfte M, et al. Trace analysis of multi-class phytohormones in Oryza sativa using different scan modes in high-resolution Orbitrap mass spectrometry : method validation, concentration levels, and screening in multiple accessions. ANALYTICAL AND BIOANALYTICAL CHEMISTRY. 2018;410(18):4527–39.
MLA
Haeck, Ashley, Herman Van Langenhove, Lies Harinck, et al. “Trace Analysis of Multi-class Phytohormones in Oryza Sativa Using Different Scan Modes in High-resolution Orbitrap Mass Spectrometry : Method Validation, Concentration Levels, and Screening in Multiple Accessions.” ANALYTICAL AND BIOANALYTICAL CHEMISTRY 410.18 (2018): 4527–4539. Print.
@article{8573256,
  abstract     = {Phytohormones are signaling and regulating metabolites involved in numerous plant processes, including growth, development, and responses to stress. Currently, the focus is on the analysis of multiple phytohormones in order to characterize crosstalk and hormone signaling networks. In this paper, representative phytohormones of the major classes are simultaneously determined in rice tissues by a generic solid-liquid extraction, followed by liquid chromatography and electrospray ionization high-resolution tandem mass spectrometry using a Q-Exactive (TM) instrument. After a thorough optimization of the sample preparation, the analytical method was fully validated toward the ultra-trace quantification of six a priori selected plant hormones using three scan modes of the quadrupole-Orbitrap instrument: full-scan high-resolution mass spectrometry, targeted single ion monitoring (t-SIM), and t-SIM followed by data-dependent tandem mass spectrometry. Overall, a similar quantitative performance was noticed for the different scan modes. The analytical method was successfully applied to measure basal phytohormone levels in six different rice accessions, comprising Oryza sativa ssp. japonica, indica, and Oryza glaberrima. Hormone concentrations were higher in shoots than in roots or at least similar. Except for a lower level of salicylic acid in shoots of O. glaberrima versus O. sativa, no other differences in hormone levels could be noticed that were dependent of the (sub)species assignment of the analyzed accessions. Making use of the benefits of full-scan high-resolution mass spectrometry, a first post-run suspect screening was performed, suggesting - based on accurate mass measurements and isotopic patterns - the possible presence of about 50 additional plant hormones in the rice tissues.},
  author       = {Haeck, Ashley and Van Langenhove, Herman and Harinck, Lies and Kyndt, Tina and Gheysen, Godelieve and H{\"o}fte, Monica and Demeestere, Kristof},
  issn         = {1618-2642},
  journal      = {ANALYTICAL AND BIOANALYTICAL CHEMISTRY},
  language     = {eng},
  number       = {18},
  pages        = {4527--4539},
  title        = {Trace analysis of multi-class phytohormones in Oryza sativa using different scan modes in high-resolution Orbitrap mass spectrometry : method validation, concentration levels, and screening in multiple accessions},
  url          = {http://dx.doi.org/10.1007/s00216-018-1112-9},
  volume       = {410},
  year         = {2018},
}

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