The invention relates to quantum-dot-based paper chips and a preparation method thereof, as well as application of the quantum-dot-based paper chips in glucose detection. The preparation method of thequantum-dot-based paper chips comprises the following steps: S1, drying porcine blood, and grinding the dried porcine blood into porcine blood powder; S2, dissolving the porcine blood, carrying out hydrothermal reaction at 150-200 DEG C for 10-18 hours, carrying out centrifuging so as to obtain liquid supernatant, filtering the liquid supernatant, and carrying out diluting so as to obtain a carbon quantum dot solution for standby application; S3, partially coating paper chips with hydrophobic matrices so as to form hydrophobic areas, and leaving the uncoated parts as hydrophilic areas; and S4, adding substrates of horseradish peroxidase and the carbon quantum dot solution into the hydrophobic areas, carrying out drying, adding a glucose oxidase solution, carrying out drying, and adding the substrates of horseradish peroxidase and the carbon quantum dot solution again so as to obtain the quantum-dot-based paper chips. The quantum-dot-based paper chips provided by the invention have activity of horseradish-like peroxidase, so that, the quantum-dot-based paper chips can be used for quickly detecting content of glucose; moreover, the detection results are more reliable and accurate, low in cost, high in application values and good in application prospects.