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Updated: Jun 8, 2026

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
Published on: July 31, 2011
Traceable amino acid analyses of proteins and peptides by isotope-dilution mass spectrometry using precolumn
Tomoya Kinumi1, Ryota Ichikawa, Hirokazu Arimoto
1Bio-Medical Standard Section, National Metrology Institute of Japan (NMIJ), National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba C-3, Ibaraki 305-8563, Japan. t.kinumi@aist.go.jp
Abstract:
We have developed an accurate and traceable quantitation method of proteins and peptides by isotope-dilution mass spectrometry with the precolumn derivatization for hydrolyzed amino acid. This method utilized N-butylnicotinic acid N-hydroxysuccinimide ester iodide as the derivatization reagent and C-30 reversed phase column for the separation. Quantitative results of porcine insulin and human serum albumin obtained from the hydrolyzed six or seven amino acids showed a good agreement, with less than 3% of the expanded uncertainties. This method allows more accurate and more robust amino acid analysis in comparison with non-labeled methods.
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