Carbon dots (CDs) have caught enormous attention owing to their distinctive properties, such as their high water solubility, tunable optical properties, and easy surface modification, which can be generally used for the detection of heavy metals and organic pollutants. Herein, nitrogen and fluorine co-doped carbon dots (NFCDs) were designed via a rapid, low-cost, and one-step microwave-assisted technique using DL-malic acid and levofloxacin. The NFCDs emitted intense green fluorescence under UV lighting, and the optical emission peak at 490 nm was observed upon a 280 nm excitation, with a high quantum yield of 21.03%. Interestingly, the spectral measurements illustrated excitation-independent and concentration-independent single-color fluorescence owing to the presence of nitrogen and fluorine elements in the surface functional groups. Additionally, the NFCDs were applied for the selective detection of Fe3+ and ascorbic acid based on the “turn-off” mode. The detection limits were determined as 1.03 and 4.22 µM, respectively. The quenching mechanisms were explored using the static quenching mechanism and the inner filter effect. Therefore, a NFCDs fluorescent probe with single color emission was successfully developed for the convenient and rapid detection of Fe3+ and ascorbic acid in environments.