Amino acid analysis by capillary electrophoresis after phenylthiocarbamylation.
Capillary electrophoresis using SDS in phosphate buffer provides high resolution and short separation time for peptide and protein hydrolysate amino acids after derivatization with phenylisothiocyanate. The phenylthiocarbamyl derivatives are quantified in the picomole and femtomole range at signal-to-noise ratios better than 3:1 (for 50 fmol) and with a linearity correlation coefficient averaging 0.9938. The migration time and peak area variabilities were on average 1.1 and 2.7%, respectively. Complete separation of all the 18 amino acids normally found in polypeptide hydrolysates is achieved in less than 30 min using 75-microm capillaries while 50-microm capillaries require less than 15 min. Analysis of peptide and protein hydrolysates in the range 10-600 residues revealed excellent agreement with the known compositions at sensitivities better by large factors than the corresponding HPLC methodology (about 20-fold) and conventional ninhydrin-based analysis (about 1000-fold).
Zahou
E
E
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
Jörnvall
H
H
Bergman
T
T
D000596:Amino Acids / Q000032:analysis* / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D006868:Hydrolysis; D010455:Peptides / Q000737:chemistry; D010670:Phenylthiourea / Q000737:chemistry; D011506:Proteins / Q000737:chemistry; D015203:Reproducibility of Results
D000596:Amino Acids; D010455:Peptides; D011506:Proteins; D010670:Phenylthiourea