An accurate, precise and sensitive method is described for the analysis of 29 polycyclic aromatic hydrocarbons (PAHs), including 19 2-6-ringed PAHs and 10 alkyl-PAHs. The method is based on an isotope dilution technique using gas chromatography/mass spectrometry (GC/MS) and available labeled PAHs as internal standards. Quality parameters were calculated with satisfactory results and 36 Spanish river sediments were analysed. Results were evaluated regarding to the sediment quality guidelines (SQGs) based on the effects range-low (ERL) and the effects range-median (ERM) values. Most analysed sediments showed a good quality, since only 7 of them exceeded ERL values, including one sample surpassing ERM values. PAH profiles were studied in order to identify PAH sources as mainly petrogenic or pyrogenic. Most samples showed petrogenic-type fingerprints, although 6 of the 11 sediments with the highest PAH concentrations (> 1000 ng/g) were classified as pyrogenic, including 4 of the 7 samples exceeding ERL values. Quality assurance was carried out by the triplicate analysis of one preanalysed river sediment without PAHs subsequently spiked at a medium (500 ng/g) and a low concentration level (10 ng/g) of each analyte. Main quality requirements for methods based on isotope dilution were accomplished. Method accuracy was 80-120% for most PAHs, method precision was <15% for all the analysed compounds and method detection limits (MDLs) were 1-3 ng/g.