The influence of high-pressure treatment on the 3 main folate species present in orange juice was assessed under conditions similar to those used in high-pressure pasteurization and sterilization using freshly squeezed orange juice and different model orange juices. The latter were used also to determine the influence of matrix components, pH, and excess ascorbic acid. Excess ascorbate strongly protected folates against pressure and heat. Pressurization at 600 MPa and 80°C affected model juice folates synergistically, and pressure increased the formation of 5,10-methenylfolate. In fresh orange juice, components other than ascorbate additionally stabilized folates, thus pressure preservation and pressure sterilization appeared very feasible.