This study aimed to evaluate the total concentration of polycyclic aromatic hydrocarbons in sediments of Iguassu River in Southern Brazil. Alongside the concentration, the amount of such compounds bioavailable was also evaluated. This is accomplished by comparing its total amount present in sediments and the amount extracted by n-butanol. The results showed that the total concentration of polycyclic aromatic hydrocarbons presented in sediment ranged from 4.49 to 58.75 μg/g. The total amount of polycyclic aromatic hydrocarbons extracted by n-butanol ranged from 1.22 to 17.07 μg/g. The use of n-butanol represents the mimetic conditions that hydrocarbons, derived from oil, could be taken up by organisms. Most of the hydrocarbons extracted by n-butanol were those with lower octanol–water partition constant, usually those with three and four rings. Compounds with more than four rings were extracted in lower or insignificant amounts. Even the hydrocarbons with lower molecular weight available may be degraded or eliminated by organisms, when accumulated. Estimating bioavailability of hydrocarbons represents what specific hydrocarbons could be available to be taken up by organisms.