We have previously detected in cardiac patients severalfold higher levels of endothelin (ET) in the pericardial fluid (PF) than in the plasma (PL). We postulated that this is due to different activities of neutral endopeptidase (NEP) in the two compartments. With approval of the ethical committee and informed consent by 32 patients (18 men, 14 women, aged 62 +/- 2 years; NYHA II-IV), PF was taken during cardiac surgery. PL samples were obtained on the day of surgery before premedication. ET was measured by radioimmunoassay after extraction (SepPakC18). NEP activity was measured by a microplate-based kinetic enzyme assay over 120 min. PF ET (78 +/- 11 pg/ml) was significantly (p < 0.05) higher than PL ET (3.38 +/- 0.48 pg/ml). The PF/PL ratio was 38 +/- 14, range 7-200. PF ET was inversely related to the NYHA state of the patients, whereas a similar relation was not found with PL ET. PL and PF ET levels did not correlate. In HPLC, the total immunoreactive ET activity co-eluted with the human ET standard. PF NEP activity (2.26 +/- 0.12 U/l) was lower (p < 0.05) than PL NEP (3.62 +/- 0.22 U/I). PL NEP was not different from that of healthy controls (3.28 +/- 0.22 U/L; n = 50). No correlation was found between NEP in either compartment and the NYHA state of the patients. ET concentration and NEP activity did not correlate in PF or PL. We conclude that ET is extremely high in the PF of cardiac patients and that this is not caused by altered NEP activity.