Selective conversion of glycerol to lactic acid requires the synergistic activation of C–H, C–O, and O–H. Herein, we report an inexpensive Fe–Cr/TiO2 catalyst that achieves lactic acid formation with a high selectivity of 60.5%. By changing the Fe/Cr ratios and reduction temperatures, the structures of Vő and Vő-OH were altered, which determined the overall oxidation–dehydration–isomerization process. The in situ ATR-IR experiments confirmed the activation of glycerol by adsorbing the primary −OH, and a series of control experiments verified that Fe was responsible for the cleavage of C–H and O–H, while CrOx catalyzed the activation of C–O and C–H. This work designs a hydrothermal catalyst with multifunctional ability, which may guide the rational catalyst design for the selective oxidation of glycerol to lactic acid. [ABSTRACT FROM AUTHOR]