The redox speciation of iron was determined during the iron fertilization LOHAFEX and for the first time, the chemiluminescence assay of filtered and unfiltered samples was systematically compared. We hypothesize that higher chemiluminescence in unfiltered samples was caused by Fe(II) adsorbed onto biological particles. Dissolved and particulate Fe(II) increased in the mixed layer steadily 6-fold during the first two weeks and decreased back to initial levels by the end of LOHAFEX. Both Fe(II) forms did not show diel cycles downplaying the role of photoreduction. The chemiluminescence of unfiltered samples across the patch boundaries showed strong gradients, correlated significantly to biomass and the photosynthetic efficiency and were higher at night, indicative of a biological control. At 150 m deep, a secondary maximum of dissolved Fe(II) was associated with maxima of nitrite and ammonium despite high oxygen concentrations. We hypothesize that during LOHAFEX, iron redox speciation was mostly regulated by trophic interactions.