Accurate detection of pH is important in pathological processes and food freshness. Developing sensors of sensitive response and visualization for pH is highly demanded. In this work, Chiral carbon dots(CCDs) was synthesized via one-pot hydrothermal process using o-phenylenediamine and L-Tryptophan, which displayed circular dichroism (CD) signals at 200–255 nm and 255–300 nm. The CCDs exhibited dual-emission peaks with blue and red emission bands when excited at 360 nm. The ratiometric signals of UV–vis absorption and fluorescence intensity of L-CDs were responsive over the pH range of 2.0 to 11.0 with significant color changes in solution. Fluorescence imaging of live cells displayed different signals related to pH in both the blue and red channels, allowing accurate measurement of the pH of the cellular environment. Furthermore, the pH test paper based on L-CDs enabled monitoring the freshness of shrimp and pork under 365 nm UV light. Therefore, L-CDs provided a multifunctional visual pH sensing platform for environmental monitoring and biosensing.