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Author Harada, Kazuo ♦ Ohyama, Yohko ♦ Tabushi, Tetsuya ♦ Kobayashi, Akio ♦ Fukusaki, Eiichiro
Source World Health Organization (WHO)-Global Index Medicus
Content type Text
Publisher Elsevier
File Format HTM / HTML
Language English
Difficulty Level Medium
Subject Domain (in DDC) Natural sciences & mathematics ♦ Chemistry & allied sciences ♦ Life sciences; biology ♦ Biochemistry ♦ Natural history of organisms ♦ Technology ♦ Medicine & health ♦ Pharmacology and therapeutics ♦ Diseases ♦ Chemical engineering ♦ Manufacture for specific uses ♦ Precision instruments & other devices
Subject Domain (in MeSH) Eukaryota ♦ Organisms ♦ Inorganic Chemicals ♦ Chemicals and Drugs ♦ Investigative Techniques ♦ Analytical, Diagnostic and Therapeutic Techniques and Equipment
Subject Keyword Discipline Biomedical Engineering ♦ Discipline Microbiology ♦ Catharanthus ♦ Metabolism ♦ Anions ♦ Electrophoresis, Capillary ♦ Spectrometry, Mass, Electrospray Ionization ♦ Journal Article ♦ Research Support, Non-u.s. Gov't
Abstract 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 mumol/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 be useful for detailed analysis of primary metabolism dynamics.
Description Country affiliation: Japan
Author Affiliation: Harada K ( Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.)
ISSN 13891723
Educational Role Student ♦ Teacher
Age Range above 22 year
Educational Use Reading ♦ Research ♦ Self Learning
Interactivity Type Expositive
Education Level UG and PG
Learning Resource Type Article
Publisher Date 2008-03-01
Publisher Place Japan
e-ISSN 13474421
Journal Journal of Bioscience and Bioengineering
Volume Number 105
Issue Number 3


Source: WHO-Global Index Medicus