Analytical Sciences

Traceable Amino Acid Analyses of Proteins and Peptides by Isotope-Dilution Mass Spectrometry Using Precolumn Derivatization Reagent

Tomoya KINUMI,* Ryota ICHIKAWA,** Hirokazu ARIMOTO,** and Akiko TAKATSU*

*Bio-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
**Graduate School of Life Sciences, Tohoku University, Aoba, Sendai 980-8577, Japan

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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