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Orbitrap Fusionを用いたCE-MSと四重極GC-MSでのアミノ酸分析の比較
澄田, 智美;島村, 繁;布浦, 拓郎
Permalink : http://hdl.handle.net/2115/81351
JaLCDOI : 10.14943/lowtemsci.79.43
KEYWORDS : メタボローム解析;安定同位体;質量分析;Metabolomics;13C tracer analysis;Stable isotope;Mass spectrometry
Abstract
微量メタボローム解析技術（アミノ酸のピバロイル/イソプロピルエステル誘導体を用いたGCMSでの分析及び質量キャリブレーション用のソフトウェアであるMassWorks を用いた解析）によるアミノ酸のアイソトポマー解析は，アミノ酸生合成経路だけでなく，前駆体となる有機酸を生産するTCA 回路等周辺の主要代謝経路の解明にも有効な手段である．しかし，この僅かなバイオマスで解析可能な手法であっても，その適用に十分なバイオマスを得ることが困難な環境微生物も存在する．また，GC-MS での解像度では，他の化合物との十分な分離が困難な場合もある．そこで，より高精度で高感度な解析手法の開発を目指し，キャピラリー電気泳動インターフェイスZipChipTM とオービトラップ搭載機種Orbitrap Fusion を用いたCE-MS での分析及び解析ソフトウェアXcalibur QualBrowser を用いた代謝解析技術の基盤構築を進めている．本報では微量メタボローム解析における，従来のGC-MS と新技術のCE-MS の両者を比較しつつ紹介する．
Isotopomer analysis for amino acids in tracer-based metabolomics is useful to identify major metabolic pathways producing amino acid precursors such as the TCA cycle in addition to amino acid biosynthetic pathways. A novel method of tracer-based metabolomics that consists of quadrupole GC-MS analysis for pivaloyl/isopropyl derivatives of amino acids and MassWorks software for identification of stable isotope probing required less amount of samples comparing with the methods available previously. However, even with the novel method, isotopomer analysis of amino acids for some of environmental microbes is not applicable because of the difficulty in obtaining sufficient biomass. Thus, we are developing an accurate and sensitive analysis by CE-MS analysis with an Orbitrap Fusion MS compound with a capillary electrophoresis interface ZipChipTM. Here, we report the comparison of current GC-MS and developing CE-MS methods to identify isotopomers of aminio acids.
Isotopomer analysis for amino acids in tracer-based metabolomics is useful to identify major metabolic pathways producing amino acid precursors such as the TCA cycle in addition to amino acid biosynthetic pathways. A novel method of tracer-based metabolomics that consists of quadrupole GC-MS analysis for pivaloyl/isopropyl derivatives of amino acids and MassWorks software for identification of stable isotope probing required less amount of samples comparing with the methods available previously. However, even with the novel method, isotopomer analysis of amino acids for some of environmental microbes is not applicable because of the difficulty in obtaining sufficient biomass. Thus, we are developing an accurate and sensitive analysis by CE-MS analysis with an Orbitrap Fusion MS compound with a capillary electrophoresis interface ZipChipTM. Here, we report the comparison of current GC-MS and developing CE-MS methods to identify isotopomers of aminio acids.
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