Using the plasmid relaxation assay, the induction of single strand breaks (SSB) and base damages was investigated in air-dried plasmid DNA irradiated under air or under vacuum, with two high LET particles. We first observed that an irradiation with 12C5+ ion produced less of both damages when performed in a vacuum rather than in the presence of air. This could be due to the presence of O2 which increases the primary radicalar effects in the latter case. Another explanation is a difference in the degree of hydration of the DNA molecules. Indeed, under vacuum only the water molecules tightly bound to DNA will persist. In contrast, in the presence of air, the outer hydration shell enhances the amount of hydroxyl radicals available for the radiolytic attack. However, no difference in the SSB induction was observed when DNA was irradiated with 36S16+ ion in the presence of air or under vacuum. This is likely due to the LET effect which partly cancels the production of radicals by recombination and increases the formation of superoxide anions in the track. Similarly, the lower induction of damage by 36S16+ irradiation in comparison with the 12C5+ ion is a consequence of the higher ionizing density for 36S16+ than for 12C5+ ions. Meanwhile, for both ions, base damages are not detected when DNA is irradiated under vacuum, whereas they are as frequent as SSB when irradiation is performed in the presence of air. Altogether, these observations support the idea that SSB and base damage are not formed by the same mechanism.