Heterocyclic amines (HCAs), which are formed during the cooking of protein-rich foods, are potent mutagens and a risk factor for human cancers. Levels of HCAs have been extensively investigated in meat products but not in fish products. Here, we report levels of HCAs in fried salmon, tuna, hake, sardine, angler fish, cod, sole, swordfish, squid, and cuttlefish. The HCA levels of some of these foods have not been previously analyzed. We employed multivariate factor-analysis tools, including principal components analysis (PCA) and partial least-squares (PLS) regression, to study the effects of cooking weight loss and levels of creatine, glucose, and free amino acids on HCA levels. The highest concentrations of mutagenic HCAs, 159.3 ng·g–1 total, where 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) accounted for 121 ng·g–1, were found in fried swordfish (cooking loss of 51.8%). These levels are higher than those generally found in fried chicken, which is typically cited as the most contaminated food item. Thus, swordfish is among the richest known sources of HCAs. The other cooked seafood items contained from 0.4 to 35.4 ng·g–1 HCAs, comparable to concentrations typically reported for meat. Chemometric analysis showed that the fish species is the most influential parameter on the formation of HCAs such as DMIP, PhIP, and norharman. Concentrations of histidine, lysine, creatine, and glucose, as well as weight loss, also influence the yield of HCAs. These results suggest that seafood is an important dietary source of HCAs. The formation of HCAs in fish is influenced by multiple factors, some of which remain unknown.