A simple and rapid HPLC method for quantification of histidine-containing dipeptides, anserine and carnosine, in animal extracts

Abstract

A simple method of HPLC for quantification of histidine-containing dipeptides (HCDP), anserine and carnosine, in animal meats was investigated. Each 10 μL of extracts prepared by heat extraction from various animal meats including beef, pork, chicken, and tuna was applied to a TSKG-2500 PWX1 column, developed with 45% acetonitrile containing 0.1% trifluoroacetic acid at a flow rate of 0.5 mL/min, and monitored with an UV detector at 210 nm. Both anserine and carnosine were eluted as an identical single peak and separated from other proteins, peptides and amino acids. Although a small amount of lysine and arginine were contained in the anserine-carnosine peak as major contaminants, these amino acids had no absorption at 210 nm. Therefore, the detected peak that showed the same retention time of anserine or carnosine was calculated as total HCDP. By using a standard curve of carnosine, total HCDP content in meat extracts was rapidly quantified within 30 min per one sample and the data obtained from this method were similar to those from an automatic amino acid analyzer.