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Development of a method for comprehensive and quantitative analysis of plant hormones by highly sensitive nanoflow liquid chromatography-electrospray ionization-ion trap mass spectrometry.
Anal Chim Acta. 2009 Aug 26; 648(2):215-25.AC

Abstract

In recent plant hormone research, there is an increased demand for a highly sensitive and comprehensive analytical approach to elucidate the hormonal signaling networks, functions, and dynamics. We have demonstrated the high sensitivity of a comprehensive and quantitative analytical method developed with nanoflow liquid chromatography-electrospray ionization-ion trap mass spectrometry (LC-ESI-IT-MS/MS) under multiple-reaction monitoring (MRM) in plant hormone profiling. Unlabeled and deuterium-labeled isotopomers of four classes of plant hormones and their derivatives, auxins, cytokinins (CK), abscisic acid (ABA), and gibberellins (GA), were analyzed by this method. The optimized nanoflow-LC-ESI-IT-MS/MS method showed ca. 5-10-fold greater sensitivity than capillary-LC-ESI-IT-MS/MS, and the detection limits (S/N=3) of several plant hormones were in the sub-fmol range. The results showed excellent linearity (R(2) values of 0.9937-1.0000) and reproducibility of elution times (relative standard deviations, RSDs, <1.1%) and peak areas (RSDs, <10.7%) for all target compounds. Further, sample purification using Oasis HLB and Oasis MCX cartridges significantly decreased the ion-suppressing effects of biological matrix as compared to the purification using only Oasis HLB cartridge. The optimized nanoflow-LC-ESI-IT-MS/MS method was successfully used to analyze endogenous plant hormones in Arabidopsis and tobacco samples. The samples used in this analysis were extracted from only 17 tobacco dry seeds (1mg DW), indicating that the efficiency of analysis of endogenous plant hormones strongly depends on the detection sensitivity of the method. Our analytical approach will be useful for in-depth studies on complex plant hormonal metabolism.

Authors+Show Affiliations

Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19646587

Citation

Izumi, Yoshihiro, et al. "Development of a Method for Comprehensive and Quantitative Analysis of Plant Hormones By Highly Sensitive Nanoflow Liquid Chromatography-electrospray Ionization-ion Trap Mass Spectrometry." Analytica Chimica Acta, vol. 648, no. 2, 2009, pp. 215-25.
Izumi Y, Okazawa A, Bamba T, et al. Development of a method for comprehensive and quantitative analysis of plant hormones by highly sensitive nanoflow liquid chromatography-electrospray ionization-ion trap mass spectrometry. Anal Chim Acta. 2009;648(2):215-25.
Izumi, Y., Okazawa, A., Bamba, T., Kobayashi, A., & Fukusaki, E. (2009). Development of a method for comprehensive and quantitative analysis of plant hormones by highly sensitive nanoflow liquid chromatography-electrospray ionization-ion trap mass spectrometry. Analytica Chimica Acta, 648(2), 215-25. https://doi.org/10.1016/j.aca.2009.07.001
Izumi Y, et al. Development of a Method for Comprehensive and Quantitative Analysis of Plant Hormones By Highly Sensitive Nanoflow Liquid Chromatography-electrospray Ionization-ion Trap Mass Spectrometry. Anal Chim Acta. 2009 Aug 26;648(2):215-25. PubMed PMID: 19646587.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of a method for comprehensive and quantitative analysis of plant hormones by highly sensitive nanoflow liquid chromatography-electrospray ionization-ion trap mass spectrometry. AU - Izumi,Yoshihiro, AU - Okazawa,Atsushi, AU - Bamba,Takeshi, AU - Kobayashi,Akio, AU - Fukusaki,Eiichiro, Y1 - 2009/07/04/ PY - 2009/04/27/received PY - 2009/06/24/revised PY - 2009/07/01/accepted PY - 2009/8/4/entrez PY - 2009/8/4/pubmed PY - 2009/9/30/medline SP - 215 EP - 25 JF - Analytica chimica acta JO - Anal Chim Acta VL - 648 IS - 2 N2 - In recent plant hormone research, there is an increased demand for a highly sensitive and comprehensive analytical approach to elucidate the hormonal signaling networks, functions, and dynamics. We have demonstrated the high sensitivity of a comprehensive and quantitative analytical method developed with nanoflow liquid chromatography-electrospray ionization-ion trap mass spectrometry (LC-ESI-IT-MS/MS) under multiple-reaction monitoring (MRM) in plant hormone profiling. Unlabeled and deuterium-labeled isotopomers of four classes of plant hormones and their derivatives, auxins, cytokinins (CK), abscisic acid (ABA), and gibberellins (GA), were analyzed by this method. The optimized nanoflow-LC-ESI-IT-MS/MS method showed ca. 5-10-fold greater sensitivity than capillary-LC-ESI-IT-MS/MS, and the detection limits (S/N=3) of several plant hormones were in the sub-fmol range. The results showed excellent linearity (R(2) values of 0.9937-1.0000) and reproducibility of elution times (relative standard deviations, RSDs, <1.1%) and peak areas (RSDs, <10.7%) for all target compounds. Further, sample purification using Oasis HLB and Oasis MCX cartridges significantly decreased the ion-suppressing effects of biological matrix as compared to the purification using only Oasis HLB cartridge. The optimized nanoflow-LC-ESI-IT-MS/MS method was successfully used to analyze endogenous plant hormones in Arabidopsis and tobacco samples. The samples used in this analysis were extracted from only 17 tobacco dry seeds (1mg DW), indicating that the efficiency of analysis of endogenous plant hormones strongly depends on the detection sensitivity of the method. Our analytical approach will be useful for in-depth studies on complex plant hormonal metabolism. SN - 1873-4324 UR - https://www.unboundmedicine.com/medline/citation/19646587/Development_of_a_method_for_comprehensive_and_quantitative_analysis_of_plant_hormones_by_highly_sensitive_nanoflow_liquid_chromatography_electrospray_ionization_ion_trap_mass_spectrometry_ DB - PRIME DP - Unbound Medicine ER -