Surface analysis by time-of-flight secondary ion mass spectrometry, X-ray photoelectron spectroscopy and scanning tunnelling microscopy has been applied to provide new insight on Mo effects on the composition and nanostructure of the passive films grown in sulfuric acid on well-controlled Fe–17Cr–14.5Ni–2.3Mo(100) austenitic stainless steel single crystal surfaces. A duplex hydroxylated oxide matrix, 1.8–1.9 nm thick, is formed with a strong partition between Cr(III) and Fe(III) in the inner and outer layers, respectively. Cr(III) is increasingly enriched by preferential iron oxide dissolution upon passivation and ageing. Ni, only present as oxide traces in the film, is enriched in the alloy underneath. Mo, mostly present as Mo(IV) in the Cr-rich inner layer prior to anodic polarisation, becomes increasingly enriched (up to 16% of cations) mostly as Mo(VI) in the Fe-rich outer layer of the passive film, with ageing promoting this effect. Metallic Mo is not significantly enriched below the passive film produced from the native oxide covered surface. Mo does not markedly impact the nanogranular morphology of the native oxide film nor its local thickness variations assigned to substrate site effects on Cr(III) enrichment. Site specific preferential passivation still takes place at the (native) oxide-covered step edges of the alloy surface, and transient dissolution remains preferentially located on the terraces. Nanostructures, possibly Mo-containing, and healing local depressions formed by transient dissolution during passivation, appear as a specific effect of the Mo presence. Another Mo effect, observed even after 20 h of passivation, is to prevent crystallisation at least in the Fe-rich outer part of the passive film where it is concentrated mostly as Mo(VI) (i.e. molybdate) species.