In this work, novel types of nitrogen-doped carbon dots (N-CDs) were prepared from citric acid and glycine (GLY) as precursors through a simple pyrolysis method. The GLY-CDs showed strong fluorescence with a fluorescence quantum yield as high as 33.34% and good water solubility. The fluorescence of GLY-CDs could be selectively quenched by iron(III) ion (Fe 3+ ) resulting in the non-fluorescent complex. Due to the high affinity of Fe 3+ to adenosine-5 0 -triphosphate (ATP), the fluorescence of the GLY-CDs in GLY-CDs-Fe 3+ could be recovered by ATP. Thereby, quantitatively fluorescent turn-on detection of ATP could be achieved. The fluorescence recovery ratio was linearly proportional to the concentration of ATP with a detection limit as low as 15.0 nM, indicating the CDs have high sensitivity. The GLY-CDs were successfully employed in the detection of ATP in serum and cell lysates. K E Y W O R D S adenosine-5 0 -triphosphate, cell lysates, glycine carbon dots, iron(III) ion 1 | INTRODUCTION Phosphate-containing metabolites, including nucleoside pyrophosphates, and phosphoproteins, are widespread in living cells.Adenosine-5 0 -triphosphate (ATP) is well known as the major energy carrier and extracellular signaling molecule in living cells, [1,2] which