Reversed-phase high-performance liquid chromatography coupled with second-order derivative spectroscopy for the quantitation of aromatic amino acids in peptides: application to hemorphins.
The characterization of aromatic amino acid-containing peptides in biological fluids or protein hydrolysates is commonly achieved using classical size-exclusion (SE) and reversed-phase (RP) high-performance liquid chromatography (HPLC) coupled with direct ultraviolet (UV) spectrometry. Here, a non-destructive quantitative determination of aromatic amino acids in peptides is developed using second-order derivative spectra obtained during RP-HPLC coupled with photodiode array detection. In this method, the free aromatic amino acids were used as standards. Sensitivity and accuracy were verified using some peptides, including bioactive hemorphins. The method was applied to determine the amounts of hemorphins present in a complex peptic bovine hemoglobin hydrolysate.
Zhao
Q
Q
Laboratoire de Genie Proteique et Cellulaire, Universite de La Rochelle, France.
Sannier
F
F
Garreau
I
I
Lecoeur
C
C
Piot
J M
JM
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis*; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D006454:Hemoglobins / Q000032:analysis* / Q000737:chemistry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D000596:Amino Acids; D006454:Hemoglobins; D010446:Peptide Fragments; C085122:hemorphin 7