Abstract: Starting from protoporphyrin a chromatographic pure hematoporphyrin dimethyl ester and its diacetyl derivative were prepared. The purification procedure with countercurrent distribution and column chromatography with alumina oxide and silica gel was described. Both compounds were identified on the basis of their IR, UV, MS, TLC, HPLC and ¹H and ¹³C NMR spectroscopic analysis. The ¹³C NMR assignments are given, which seem to be useful for the identification of different porphyrins.