TY - JOUR
T1 - Improved detection of di-peptides by liquid chromatography-tandem mass spectrometry with 2,4,6-trinitrobenzene sulfonate conversion
AU - Nakashima, Eduardo M.N.
AU - Qing, Hu Qiang
AU - Tanaka, Mitsuru
AU - Matsui, Toshiro
N1 - Funding Information:
This work was supported by grant aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan (no. 22248014) to TM. The authors thank Yuri Iwaihara and Chihiro Hashimoto of Kyushu University for their technical support.
PY - 2013
Y1 - 2013
N2 - 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.
AB - 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.
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U2 - 10.1271/bbb.130448
DO - 10.1271/bbb.130448
M3 - Article
C2 - 24096674
AN - SCOPUS:84886679444
SN - 0916-8451
VL - 77
SP - 2094
EP - 2099
JO - Bioscience, Biotechnology and Biochemistry
JF - Bioscience, Biotechnology and Biochemistry
IS - 10
ER -