High-speed analysis of Dabsyl amino acids in collagen by UHPLC

Collagen, one of the proteins has been recently attracting attention from the viewpoint of health and beauty, since collagen is a main constituent of corium which gives skin vitality and keeps skin soft without wrinkles. Amino acid structure of collagen is known as the arrangement of glycine sequencing at every 3 residue and contains other amino acids such as proline, hydroxyproline and alanine. In addition as specific amino acids for collagen, there are hydroxyproline and hydroxylysine. In order to measure the constituent amino acids of proteins or peptides such as collagen, sample preparation such as hydrolysis and so on needs to be implemented. The DAB-Label kit available from Jasco for amino acid derivatization using Dabsyl chloride as reagent includes the tools and reagents for vapor-phase hydrolysis by hydrochloric acid for proteins or peptides, and by using the DAB-Label kit, collagen can be hydrolyzed and amino acids can be derivatized with Dabsyl chloride in order to analyze the constituent amino acids of collagen.

In this report, collagen was hydrolyzed with the DAB-Label kit and derivatized amino acids were separated by Ultra High-performance Liquid Chromatography (UHPLC) using a UV detector.