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Use of ultra‐performance liquid chromatography/time‐of‐flight mass spectrometry with nozzle‐skimmer fragmentation for comprehensive quantitative analysis of secondary metabolites in Arabidopsis thaliana
Author(s) -
Sugimoto Takanori,
Bamba Takeshi,
Izumi Yoshihiro,
Nomura Hironari,
Shiina Takashi,
Fukusaki Eiichiro
Publication year - 2011
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201100552
Subject(s) - chromatography , mass spectrometry , time of flight mass spectrometry , chemistry , fragmentation (computing) , time of flight , analytical chemistry (journal) , ionization , organic chemistry , biology , ion , ecology
Abstract This study sought to develop techniques for LC/MS‐based metabolomics and to verify that an MS/MS spectral tag (MS2T) could be used in practical secondary metabolite profiling. The retention time (RT), precursor ions, and fragment ions generated by nozzle‐skimmer fragmentation were determined using ultra‐performance liquid chromatography/time‐of‐flight mass spectrometry (UPLC/TOF‐MS) and compared with the MS2T. A standard mix was analyzed with UPLC/TOF‐MS under the same conditions as were used to construct the MS2T. The difference in RT for the standards was less than 0.15 min and the average RSD was about 2.8%, suggesting that the analysis was highly repeatable. Both precursor ions and fragment ions were observed when the cone voltage was 75 V. Experimental data and fragmentation pattern in the MS2T annotation list were highly similar. Wild‐type and cas‐1 mutant Arabidopsis thaliana samples treated with an elicitor were analyzed using UPLC/TOF‐MS. Sixty‐five peaks were successfully annotated. Fragment ions were observed with nozzle‐skimmer fragmentation in 50 of 65 (77%) peaks. The reliability of annotation may have increased as a result of fragment ions. Results of multivariate analysis suggested that cas‐1 was related to induction of the biosynthesis of these flavonoids. The devised method facilitated practical secondary metabolite profiling.

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