The invention relates to the technical field of preparation of carbon nanomaterials and provides a preparation method of nitrogen-doped fluorescent carbon dots. The preparation method comprises the following steps: 1, a carbon source is broken, ground, sieved by a 200-mesh sieve and dried in an oven; 2, the carbon source dried by the oven is weighed and placed into a 50-ml beaker, then an organic solvent is added, and a mixture is stirred sufficiently and subjected to ultrasonic processing for 0.8-1.2 h; 3, the mixture prepared in the step 2 is placed in a 50-ml hydrothermal kettle to have a hydrothermal reaction for 2-12 h, and the temperature is controlled at 60-180 DEG C; 4, after step 3, a suspension is taken out and placed into a high-speed desk centrifuge, supernatant liquor is taken out after 10-20 min of centrifugation, and hydrosol of the fluorescent carbon dots is obtained; 5, the hydrosol, obtained in the step 4, of the fluorescent carbon dots is placed in rotary evaporation equipment, the organic solvent is removed, and the target product fluorescent carbon dots are prepared. The preparation method of the nitrogen-doped fluorescent carbon dots has the characteristics of green and environment-friendly reaction conditions, easiness in control and easiness in mass production and preparation and is expected to be widely applied to the fields of photoelectric devices, bio-imaging, detection, sensing and the like.