Development of a capillary electrophoresis-mass spectrometry method using polymer capillaries for metabolomic analysis of yeast.
Metabolomics is an emerging field in analytical biochemistry, and the development of such a method for comprehensive and quantitative analysis of organic acids, carbohydrates, and nucleotides is a necessity in the era of functional genomics. When a concentrated yeast extract was analyzed by CE-MS using a successive multiple ionic-polymer layer (SMIL)-coated capillary, the adsorption of the contaminants on the capillary wall caused severe problems such as no elution, band-broadening, and asymmetric peaks. Therefore, an analytical method for the analysis of anionic metabolites in yeast was developed by pressure-assisted CE using an inert polymer capillary made from poly(ether etherketone) (PEEK) and PTFE. We preferred to use the PEEK over the PTFE capillary in CE-MS due to the easy-to-use PEEK capillary and its high durability. The separation of anionic metabolites was successfully achieved with ammonium hydrogencarbonate/formate buffer (pH 6.0) as the electrolyte solution. The use of 2-propanol washing after every electrophoresis run not only eliminated wall-adsorption phenomena, but allowed for good repeatability to be obtained for migration times in the metabolomic analysis.
Tanaka
Yoshihide
Y
Human Stress Signal Research Center, National Institute of Advanced Industrial Science and Technology, Ikeda, Osaka, Japan. yoshihide-tanaka@aist.go.jp
Higashi
Tetsuji
T
Rakwal
Randeep
R
Wakida
Shin-ichi
S
Iwahashi
Hitoshi
H
D001577:Benzophenones; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D007659:Ketones; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D011092:Polyethylene Glycols; D011108:Polymers; D011138:Polytetrafluoroethylene; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D012822:Silicon Dioxide
D001577:Benzophenones; D007659:Ketones; D011108:Polymers; C063834:polyetheretherketone; D011092:Polyethylene Glycols; D012822:Silicon Dioxide; D011138:Polytetrafluoroethylene