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The oxidation of highly toxic arsenite (As(III)) was studied using humic acid-coated magnetite nanoparticles (HA-MNP) as a photosensitizer. Detailed characterization of the HA-MNP was carried out before and after the photoinduced treatment of As(III) species. Upon irradiation of HA-MNP with 350 nm light, a portion of the As(III) species was oxidized to arsenate (As(V)) and was nearly quantitatively removed from the aqueous solution. The separation of As(III) from the aqueous solution is primarily driven by the strong adsorption of As(III) onto the HA-MNP. As(III) removals of 40–90% were achieved within 60 min depending on the amount of HA-MNP. The generation of reactive oxygen species (•OH and 1O2) and the triplet excited state of HA-MNP (3HA-MNP*) was monitored and quantified during HA-MNP photolysis. The results indicate 3HA-MNP* and/or singlet oxygen (1O2) depending on the reaction conditions are responsible for converting As(III) to less toxic As(V). The formation of 3HA-MNP* was quantified using the electron transfer probe 2,4,6-trimethylphenol (TMP). The formation rate of 3HA-MNP* was 8.0 ± 0.6 × 10−9 M s−1 at the TMP concentration of 50 µM and HA-MNP concentration of 1.0 g L−1. The easy preparation, capacity for triplet excited state and singlet oxygen production, and magnetic separation suggest HA-MNP has potential to be a photosensitizer for the remediation of arsenic (As) and other pollutants susceptible to advanced oxidation.