Little is known about the extent to which Atlantic eels coming from different European rivers converge on the same spawning site. Our aim was to evaluate the spatial homogeneity of eel spawning area(s) with an otolith microchemistry approach. This work compared the elemental signatures of otolith's core region of Anguilla anguilla leptocephali caught in the Sargasso Sea in 2007 with those of glass eels and elvers sampled in European estuaries during 2006, 2008 and 2009. Using laser ablation inductively coupled plasma mass spectrometry, the same annular ablation trajectory along the first feeding mark was applied on otoliths of glass eels, elvers and leptocephali. The concentrations of thirteen isotopes in the otoliths of glass eels ⁄ elvers did not vary among three annual cohorts collected in eleven European locations. However, otolith elemental fingerprints of leptocephali differed significantly from glass eels otolith's signature. Although the mechanisms that regulate the differences in trace element signatures among leptocephali and glass eels ⁄ elvers are unknown, we propose that the sampled glass eels ⁄ elvers were born in a spawning site or region where favourable transport and ⁄ or feeding conditions occurred. Conversely, the leptocephali may have been sampled in a less favourable region in the Sargasso Sea, with a low probability of reaching continental growth areas.