Development of a New LC-MS/MS Method for the Quantification of Keto Acids

• NOGUCHI Kazuyoshi

  Fundamental Technology Labs., Institute For Innovation, Ajinomoto Co., Inc.

• MIZUKOSHI Toshimi

  Fundamental Technology Labs., Institute For Innovation, Ajinomoto Co., Inc.

• MIYANO Hiroshi

  Fundamental Technology Labs., Institute For Innovation, Ajinomoto Co., Inc.

• YAMADA Naoyuki

  Fundamental Technology Labs., Institute For Innovation, Ajinomoto Co., Inc.

抄録

Keto acids are known to be key intermediates in various metabolic pathways. The development of an accurate method for the determination of keto acids is therefore important for diagnosing metabolic disorders as well as elucidating cellular metabolic processes in the TCA cycle, glycolysis and amino acid biosynthesis. In this study, we have developed a comprehensive and reliable LC-MS/MS method for the analysis of biological samples using a pre-column derivatization process. Ten keto acids, including α- and β-keto acids, were converted to the corresponding O-(2,3,4,5,6-pentafluorobenzyl)oxime derivatives and analyzed by LC-MS/MS. Oxaloacetic acid, which is generally considered to be unstable, was also successfully derivatized under mild reaction conditions. The pretreatment procedure used in this study was simple and did not require any difficult extraction or evaporation processes. The separation and detection of the derivatized keto acids was achieved using an LC-MS/MS system in multiple reaction monitoring mode. This newly developed method was applied to the analysis of keto acids in rat plasma, and showed good reproducibility (1.1–4.7% as CV) and recovery (96–109%) rates. This method also exhibited a low limit of detection in the range of 0.01–0.25 μM and good linearity (r² > 0.997) over a wide concentration range of up to 300 μM. Based on these performance characteristics, this method could be readily applied to the comprehensive analysis of keto acids in biological samples.

収録刊行物

• CHROMATOGRAPHY

  CHROMATOGRAPHY 35 (3), 117-123, 2014

  クロマトグラフィー科学会

キーワード

• Keto acid
• Quantification method
• O-(2,3,4,5,6-Pentafluorobenzyl)oxime
• LC-MS/MS

詳細情報

• CRID: 1390001205745184000
• NII論文ID: 130004715563
• NII論文ID: 40020306341
• NII書誌ID: AA1137755X
• DOI: 10.15583/jpchrom.2014.017
• ISSN: 13483315
• ISSN: 13428284
• NDL書誌ID: 025991204
• Web Site: http://id.ndl.go.jp/bib/025991204
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I025991204
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可