The invention relates to a carbon quantum dot fluorescent-labeled material with coke powder serving as a carbon source and a preparation method of the carbon quantum dot fluorescent-labeled material. According to the technical scheme, the mass ratio of the coke powder to concentrated H2SO4 to concentrated HNO3 is 0.1: (9-15):(3-5), firstly, the coke powder is put in a reaction kettle, then mixed acid of concentrated H2SO4 and concentrated HNO3 is added, the mixture is stirred for 8-12 hours at a temperature of 85-105 DEG C, and cooling and solid-liquid separation are performed; the pH value is regulated to 7-11, and a brown black liquid is obtained; the brown black liquid, acetone and isopropanol are stirred and centrifugalized, and a lower-layer solution is obtained; the brown black liquid and absolute ethyl alcohol are stirred and centrifugalized, and a carbon quantum dot solution is obtained; the carbon quantum dot solution is dried, and the carbon quantum dot fluorescent-labeled material with the coke powder serving as the carbon source is obtained. The carbon quantum dot fluorescent-labeled material with the coke powder serving as the carbon source has the characteristics that the reaction temperature is low, the process is simple, the cost is low, separation and purification are facilitated, the yield of carbon quantum dots is high, and industrialization is facilitated; the prepared carbon quantum dot fluorescent-labeled material with the coke powder serving as the carbon source has good optical performance and low cytotoxicity.