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Eduardo M N NAKASHIMA, Hu-Qiang QING, Mitsuru TANAKA, Toshiro MATSUI, Improved Detection of Di-peptides by Liquid Chromatography-Tandem Mass Spectrometry with 2,4,6-Trinitrobenzene Sulfonate Conversion, Bioscience, Biotechnology, and Biochemistry, Volume 77, Issue 10, 23 October 2013, Pages 2094–2099, https://doi.org/10.1271/bbb.130448
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Abstract
Highly sensitive detection of small peptides at the pm level was achieved by liquid chromatography-multiple reaction monitoring-tandem mass spectrometry (LC-MRM-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.