More rapid triaging of patients suspected for acute myocardial infarction (AMI) is needed to facilitate early initiation of appropriate therapy in patients with AMI and to exclude low-risk patients who can safely be sent home. Heart-type fatty acid-binding protein (H-FABP) is established to be the earliest available plasma marker for AMI diagnosis. There is a paucity of data on circulating levels of H-FABP in the general population. In this study we applied a H-FABP immunoassay with improved analytical performance to assess plasma H-FABP reference values in a relatively large group (n = 443) of healthy subjects aged 18-69 y. Mean (±SD) plasma H-FABP concentration was 1.7 ± 0.9 ng/mL, with somewhat higher values found in males than in females. Plasma H-FABP also increases with age, as previously reported. The 99th centile was obtained for subjects aged 41-69 years and amounted to 5.6 ng/mL.