Antler has the benefits of increasing kidney yang, nourishing essence and blood, and strengthening tendons and bones. In the face of the different types of antler, screen out a representative ingredient that represents the overall quality of the herb from the comprehensive evaluation of quality, and therapeutic efficacy and recognize its rapid detection has become the key to the initial rapid evaluation of the quality of antler. In recent years, hydroxyproline (Hyp) has been recognized as one of the index components of antler quality evaluation. Molecularly imprinted polymer membranes (MIPs) has found significant application in the field of chemical sensing. In this study, we prepared MIPs by self-assembly on ferrocene (Fc)-multi-walled carbon nanotubes (MWCNTs) modified glassy carbon electrode (GCE). Thus, we developed a molecularly imprinted electrochemical sensor for highly selective, sensitive, and rapid detection of the Hyp. The sensor had a detection limit of 9.0 × 10-3 μg/mL within a range of 1.0 × 10-2 ~ 1.0 × 103 μg/mL (S/N = 3). Using the sensor in actual samples resulted in recoveries ranging from 97.45 ~ 107.70%, RSD of 0.53 ~ 1.28%. This work provides a novel approach for the initial screening evaluation of the quality of Chinese medicinal materials.