Traceable amino acid analyses of proteins and peptides by isotope-dilution mass spectrometry using precolumn derivatization reagent.
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.
Kinumi
Tomoya
T
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. t.kinumi@aist.go.jp
Ichikawa
Ryota
R
Arimoto
Hirokazu
H
Takatsu
Akiko
A
D000596:Amino Acids / Q000032:analysis*; D000818:Animals; D002417:Cattle; D006801:Humans; D006868:Hydrolysis; D007202:Indicators and Reagents; D007328:Insulin / Q000032:analysis; D007554:Isotopes; D013058:Mass Spectrometry; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D012709:Serum Albumin / Q000032:analysis; D013388:Succinimides; D013552:Swine
D000596:Amino Acids; D007202:Indicators and Reagents; D007328:Insulin; D007554:Isotopes; D010455:Peptides; D011506:Proteins; D012709:Serum Albumin; D013388:Succinimides; C001426:N-hydroxysuccinimide