Plasma concentrations of endothelin (ET)-1 and -3 were determined simultaneously in dogs with various pathophysiological conditions, because these peptides may display different pharmacological profiles. The study pays special attention to the characterization of plasma ET immunoreactivity (ET-IR), using high-pressure liquid chromatography (HPLC) analysis with off-line detection by radio-immunoassay (RIA). In most sick dogs evaluated total plasma ET-1-IR concentration did not differ from that of healthy dogs. However, HPLC analysis of their total plasma ET-1-IR revealed distinct ET-IR profiles. Big-ET-1, which is barely detectable in control dogs, does represent the predominant ET in sick dogs. Regardless of the pathophysiological conditions, considerable amounts of high-molecular weight ET-1-IR, most likely aggregated ET-material, was found consistently. With respect to ET-3, we constantly observed moderately increased concentrations, though no major difference of molecular pattern was evident between healthy and sick dogs. The data show a distinct regulation of ET-1 and ET-3 in dogs. Furthermore, specific molecular forms of ET-IR were found to occur in various diseases. The endothelins may therefore prove to be of diagnostic importance in the pathophysiology of vascular diseases.