Improved detection of di-peptides by liquid chromatography-tandem mass spectrometry with 2,4,6-trinitrobenzene sulfonate conversion

Eduardo M.N. Nakashima, Hu Qiang Qing, Mitsuru Tanaka, Toshiro Matsui

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

Highly sensitive detection of small peptides at the pM level was achieved by liquid chromatography-multiple reaction monitoring-tandem mass spectrometry (LCMRM- MS/MS) in combination with the 2,4,6-trinitrobenzene sulfonate (TNBS) conversion technique. Six di-peptides having Tyr at the C-terminal (i.e., Gly-Tyr, Val-Tyr, Met-Tyr, Glu-Tyr, Lys-Tyr and His-Tyr) were subjected to the TNBS-MRM analysis in this study. The TNBS conversion conditions of pH 8.0, 30 °C and 60- min incubation enabled the di-peptides to be successfully converted to a trinitrophenyl (TNP) form with the mass increment of +212 Da. The proposed TNBS-MRM method enabled di-peptide detection that was highly improved by a factor of 3-55 in signal-to-noise ratio due to increased hydrophobicity by the induced TNP moiety. The method also permitted highly sensitive detection of di-peptides with a detection limit of >54 pM (>1:35 fmol/injection), achieving high reproducibility (<5%coefficient of variation) and rapidity (<30 min) by LC-MRM-MS/MS.

Original languageEnglish
Pages (from-to)2094-2099
Number of pages6
JournalBioscience, Biotechnology and Biochemistry
Volume77
Issue number10
DOIs
Publication statusPublished - 2013

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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