Simultaneous RP-HPLC-DAD determination of dansyl amino acids in chemically treated human hair

Pece Sherovski, Marina Stefova, Natasha Ristovska


A simple, rapid and reliable RP-HPLC-DAD method for the analysis of 19 amino acids in the protein hydrolysate of chemically treated hair is developed. The pre-column dansylation and RP-HPLC separation were optimized and peak resolution was improved as compared to previously published HPLC methods. The developed method showed excellent linearity for all tested amino acids in the concentration range 0.05-0.5 mmol/l. The obtained results show that the concentration of all amino acids detected in bleached hair decreases with increasing the concentration of hydrogen peroxide, this is most significant for cystine, compared to virgin hair. Higher amounts of the typical amino acids for keratin have been measured after exposure of hair to protein treatments. The method has been shown as very useful for amino acids analysis in a protein hydrolisate and it can be also used for establishing the amino acid profiles of other protein samples using this optimized procedure.


HPLC; dansyl amino acids; protein hydrolisate; hair proteins; bleaching, protein treatments;


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