The luminescent carbon dots with peroxidase mimicking property had attracted considerable attention in biomedical field. In this work, iron-doped carbon dots (Fe-CDs) were prepared by one-pot hydrothermal method with 5, 10, 15, 20-tetra (4-borate phenyl)−21H, 23H-porphyrin Fe (II) (Fe-TBPP) as precursor. The obtained Fe-CDs emitted intense blue luminescence under ultraviolet light irradiation. Moreover, the Fe-CDs exhibited remarkable peroxidase mimicking property, which can efficiently catalyze the oxidation of 3, 3′, 5, 5′-tetramethylbenzidine (TMB) into blue ox-TMB in the presence of hydrogen peroxide (H2O2). More importantly, the emission of Fe-CDs could be gradually quenched with the addition of H2O2. Based on these phenomena, a new optical dual-mode (colorimetric and fluorometric) method for the detection of H2O2 and glucose was successfully established. The detection limits of glucose were calculated to be 3.86 and 7.27 μM (S/N = 3) respectively based on the colorimetric and fluorometric methods. Furthermore, we combined this dual-mode detection method with smartphone imaging. The colorimetric and fluorescent images were collected by recognition software of smartphone, which were then transformed into the corresponding HSL values for quantitative determination of glucose. Finally, the dual-mode approach based on Fe-CDs was used for the detection of glucose content in human serum, demonstrating the potential application of carbon dots in the biological area.