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This paper deals with a study of the mechanism of hydrodeoxygenation (HDO) at 0.1 MPa and 300–350 °C of 2-methyltetrahydrofuran (2-MTHF) over a Ni2P/SiO2 catalyst. The study was conducted using in situ Fourier transform infrared (FTIR) spectroscopy to monitor the adsorbed species during reaction and in situ X-ray absorption fine structure (XAFS) to probe the oxidation state of the Ni component. The work is relevant to the upgrading of bio-oil derived from the pyrolysis of biomass. It was deduced that the initial interaction of the 2-MTHF was with OH groups on the catalyst surface, followed by adsorption on Ni sites. Ring-opening of the tetrahydrofuran led to the formation of the main products of the reaction, n-pentane by HDO and n-butane + CO by decarbonylation. The CO band appeared with a slight delay in the FTIR spectrum because the reaction to form n-pentane was favored and turned over about four times before one n-butane + CO was formed.