The synthesis of dual-emissive carbon dots (CDs) with a longer emission wavelength by using a facile strategy is of great importance for the fabrication of ratiometric fluorescent nanoprobes. Herein, red/green dual-emissive carbon dots (RGDE CDs) were synthesized in one step using 2,5-diaminotoluene sulfate (DATS) as a carbon source. The as-prepared RGDE CDs not only exhibited dual emission fluorescence peaks (525 nm, 603 nm) at the single excitation wavelength of 370 nm, but also possessed good water solubility and excellent fluorescence stability. Moreover, the as-prepared RGDE CDs could be directly utilized as a ratiometric fluorescent probe for the determination of trace ONOO- due to the electron transfer process from ONOO- to the excited RGDE CDs. Under optimal conditions, the linear range was 0.03-60 μM with the limit of detection of 11.6 nM. Importantly, this RGDE CD probe could be applied for the detection of intracellular ONOO- with excellent biocompatibility and cellular imaging capability, indicating great promise in biomedical applications.