Abstract Various groups have reported Fe L23 X-ray absorption spectra (XAS) of magnetite (Fe3O4), each claiming to be that of magnetite, but which contradict each other. Here we report an XAS study of two kinds of magnetite: one is biogenic magnetite nanocrystals extracted from the magnetotactic bacterium Magnetovibrio blakemorei strain MV-1; the other is synthetic, abiogenically produced nano-magnetite. We see significantly different XAS spectra of these two materials. Only when the abiogenic magnetite was reduced under H2 did it give the same spectrum as the biogenic sample. Extensive heating of the biogenic magnetite in air produced spectra similar to that of the abiogenic magnetite. These two spectra are typical of the range of published Fe L23 spectra of magnetite. X-ray diffraction confirmed that the biogenic material is stoichiometric Fe3O4, and showed that the as-received or partly reduced abiogenic material is a non-stoichiometric oxide, intermediate between magnetite and maghemite (γ-Fe2O3). When the membrane which surrounds magnetosome chains was intact, the biotic magnetite single crystals were surprisingly resistant to oxidation. This study clarifies a significant confusion existing in the literature as to the correct Fe L23 X-ray absorption spectra of magnetite and maghemite.