Establishment of a Two-Dimensional HPLC-MS/MS Method Combined with DCI/D₂O Hydrolysis for the Determination of Trace Amounts of D-Amino Acid Residues in Proteins

• ISHIGO Shoto

  Graduate School of Pharmaceutical Sciences, Kyushu University

• NEGISHI Eiichi

  Shiseido Irica Technology Inc.

• MIYOSHI Yurika

  Graduate School of Pharmaceutical Sciences, Kyushu University

• ONIGAHARA Hirohisa

  Graduate School of Pharmaceutical Sciences, Kyushu University

• MITA Masashi

  Shiseido Co., Ltd.

• MIYAMOTO Tetsuya

  Department of Pharmaceutical and Life Sciences, Kitasato University

• MASAKI Haruhiko

  Department of Biotechnology, The University of Tokyo

• HOMMA Hiroshi

  Department of Pharmaceutical and Life Sciences, Kitasato University

• UEDA Tadashi

  Graduate School of Pharmaceutical Sciences, Kyushu University

• HAMASE Kenji

  Graduate School of Pharmaceutical Sciences, Kyushu University

書誌事項

タイトル別名

• Establishment of a Two-Dimensional HPLC-MS/MS Method Combined with DCl/D₂O Hydrolysis for the Determination of Trace Amounts of D-Amino Acid Residues in Proteins
• Establishment of a two-dimensional HPLC-MS/MS method combined with DCl/D2O hydrolysis for the determination of trace amounts of d-amino acid residues in proteins

抄録

A two-dimensional (2D) HPLC-MS/MS method has been established for the determination of trace amounts of D-amino acid residues in proteins. D-Amino acid residues are now increasingly recognized as biomarkers of diseases and key moieties for the regulation of protein structures/functions, therefore, a sensitive analytical method is highly required. However, non-negligible amounts of D-amino acids are produced by chemical racemization during the hydrolysis of proteins, and a sensitive determination of D-amino acid residues is normally difficult. In the present study, DCl/D₂O hydrolysis is adopted and the produced deuterated D-amino acids are distinguished from naturally-occurring D-amino acids in the proteins using the 2D-HPLC-MS/MS system. D-Ala, D-Asp, D-Glu, D-Pro and D-Ser (frequently observed D-amino acids in higher animals) were selected as the targets, and the sensitive determination (around 1% or less of the L-forms) could be carried out. As an application, the D-amino acid residues in ovalbumin (OVA) were determined, and the presence of a significant amount of D-Ser (1.8% of L-Ser) was demonstrated.

収録刊行物

• CHROMATOGRAPHY

  CHROMATOGRAPHY 36 (2), 45-50, 2015

  クロマトグラフィー科学会

キーワード

• Enantiomer separation
• D-Amino acid residue
• 2D-HPLC-MS/MS

詳細情報

• CRID: 1390282680721229312
• NII論文ID: 130005098795
• NII書誌ID: AA1137755X
• DOI: 10.15583/jpchrom.2015.017
• ISSN: 13483315
• ISSN: 13428284
• NDL書誌ID: 026704658
• Web Site: http://id.ndl.go.jp/bib/026704658
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I026704658
• Web Site: https://www.jstage.jst.go.jp/article/jpchrom/36/2/36_2015.017/_pdf
• 本文言語コード: en
• データソース種別:
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  • NDL
  • Crossref
  • CiNii Articles
  • KAKEN
• 抄録ライセンスフラグ: 使用不可