A new-style white pepper derived dual-emission carbon dots (CDs) with a quantum yield of 10.4% was designed and facile constructed with one-pot solvothermal method. The green emission (520 nm) had an efficient and special "turn-on" fluorescence sensing of coenzyme A (CoA) with the aid of Cu, while red emission (668 nm) barely changed and worked as reference. In the concentration range (0-150 µM), relative fluorescence intensity ratios (F/F) showed excellent linear correlation with concentrations of CoA, and detection limit was as low as 8.75 nm. Moreover, the strategy has been successfully applied for label-free detection of CoA in real pig liver samples with good recoveries (93.3-108.0%). Notably, the synthesized CDs had durable fluorescence, low cytotoxicity, and good biocompatibility for cellular imaging, which demonstrated wide and promising applicability for biosensing and bioimaging in the future.