Quantitative Analysis of Anionic Metabolites for Cathamnthus roseus by Capillary Electrophoresis Using Sulfonated Capillary Coupled with Electrospray lonization-Tandem Mass Spectrometry(METHODS)
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概要
- 論文の詳細を見る
This paper describes a practical method to quantify anionic metabolites using contemporary capillary electrophoresis-electrospray ionization-tandem mass spectrometry(CE-ESI-MS/MS). The use of sulfonated capillary permitted a simultaneous analysis of sugar phosphates, organic acids, nucleotides and coenzyme A compounds with only one CE condition. This capillary also improved the repeatability and sensitivity. MS/MS with multiple reaction monitoring(MRM) detection was utilized to achieve significant selectivity and sensitivity. Under optimized CE-ESI-MS/MS system, the detection limits of 53 metabolites at signal-to-noise ratio of 3 were between 0.040 and 8.8 μmol/l. The relative standard deviations(RSDs) for the majority anionic metabolites were better than 0.5% for migration times, and better than 10.0% for peak areas(n=6). Matrix effects by contaminants in sample solution in CE-ESI-MS/MS analysis were removed dramatically by the sample preparation method with liquid-liquid fractionation and ultrafiltration procedure. Furthermore, the developed method was successfully applied to determine anionic metabolites in a cultured cell of Catharanthus roseus. Accumulation of some metabolites including shikimate, malate, and sedoheptulose 7-phosphate by elicitation of methyl jasmonate was observed. The result would show shikimate, tricarboxylic acid and pentose phosphate pathways were activated. Our method will b useful for detailed analysis of primary metabolism dynamics.
- 社団法人日本生物工学会の論文
- 2008-03-25
著者
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Fukusaki Eiichiro
Department of Biotechnology, Graduate School of Engineering, Osaka University
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Harada Kazuo
Department Of Biotechnology Graduate School Of Engineering Osaka University:(present Office)departme
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KOBAYASHI AKIO
Department of Biotechnology, Graduate School of Engineering, Osaka University
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KAWABE Akira
Department of Biotechnology, Graduate School of Engineering, Osaka University
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Kobayashi A
National Inst. Advanced Industrial Sci. And Technol. Ibaraki Jpn
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Kobayashi Akio
Dep. Of Biotechnology Graduate School Of Engineering Osaka Univ.
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Kobayashi A
Laboratory Of Bioresources Chemistry Faculty Of Agriculture Okayama University
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Harada Kazuo
Applied Environmental Biology Graduate School Of Pharmaceutical Sciences Osaka University
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Kobayashi A
Ochanomizu Univ. Tokyo Jpn
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Kobayashi A
Dep. Of Biotechnology Graduate School Of Engineering Osaka Univ.
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Kawabe Akira
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Ohyama Yohko
Department of Biotechnology, Graduate School of Engineering, Osaka University
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Tabushi Tetsuya
Department of Biotechnology, Graduate School of Engineering, Osaka University
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Fukusaki Eiichiro
Division Of Advanced Science And Biotechnology Graduate School Of Engineering Osaka University
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Fukusaki Eiichiro
Dep. Of Biotechnology Graduate School Of Engineering Osaka Univ.
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Fukusaki Ei-ichiro
大阪大学大学院
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Ohyama Yohko
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Hirata Kazumasa
Dep. Of Applied Environmental Biology Graduate School Of Pharmaceutical Sciences Osaka Univ.
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Fukusaki E
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Fukusaki Eiichiro
Department Of Biotechnology Faculty Of Engineering Osaka University
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Fukusaki Eiichiro
Dept. Mat. Life Sci. Osaka Univ
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Harada Kazuo
Department Of Applied Environmental Biology Graduate School Of Pharmaceutical Sciences Osaka Univers
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Tabushi Tetsuya
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Kobayashi Akio
Department Of Biotechnology Graduate School Of Engineering Osaka University
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Tabuchi Tetsuya
Department of Biotechnology, Graduate School of Engineering, Osaka University
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KOBAYASHI Akio
Department of Biochemistry, Shinshu University School of Medicine
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KOBAYASHI Akio
Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University
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