A method is described for separation of carnosine and anserine and of the amino acids constituting them by means of thin-layer chromatography. For quantitative estimation the amino acids and dipeptides were treated with 7-chlorine-4-nitrobenzeno-2-hydroxy-1,3-diazol (NBD-chloride) and subsequently separated by chromatography on silica gel and estimated by absorption at 456-470 nm. Systems of solvents were developed for separation of both free and chemically modified amino acids and dipeptides; optimal conditions were chosen for preparation of tissue extracts and their treatment with NBD-chloride. By this method carnosine and anserine could be stimated in extract from 0.02-0.2 mg of muscular tissue. The time required for the analyses was about 3-5 hrs.