Nuclear inelastic scattering (NIS) of synchrotron radiation, also known as nuclear resonant vibrational spectroscopy (NRVS), has been shown to provide valuable insights into metal-centered vibrations at Mössbauer-active nuclei. We present a study of the iron-centered vibrational density of states (VDOS) during the first step of the Gif-type oxidation of cyclohexene with a novel trinuclear Fe 3 (μ 3 -O) complex as catalyst precursor. The experiments were carried out on shock-frozen solutions for different combinations of reactants: Fe 3 (μ 3 -O) in pyridine solution, Fe 3 (μ 3 -O) plus Zn/acetic acid in pyridine without and with addition of either oxygen or cyclohexene, and Fe 3 (μ 3 -O)/Zn/acetic acid/pyridine/cyclohexene (reaction mixture) for reaction times of 1 min, 5 min, and 30 min. The projected VDOS of the Fe atoms was calculated on the basis of pseudopotential density functional calculations. Two possible reaction intermediates were identified as [Fe (III) (C 5 H 5 N) 2 (O 2 CCH 3 ) 2 ] + and Fe (II) (C 5 H 5 N) 4 (O 2 CCH 3 ) 2 , yielding evidence that NIS (NRVS) allows to identify the presence of iron-centered intermediates also in complex reaction mixtures.