In this study, we constructed a highly sensitive and selective electrochemical sensing strategy for L-ascorbic acid (AA) based on a covalent organic framework (COF)-loading non-noble transition metal Co ion and macrocyclic cationic pillar[6]arene (CP6) nanocomposite (CP6-COF-Co). The COF plays a crucial role in anchoring the Co ion according to its crystalline porous and multiple coordination sites and has an outstanding performance for building an electrochemical sensing platform based on a unique two-dimensional structure. Accordingly, the transition-metal Co ion can be successfully anchored on the framework of COF and shows strong catalytic activity for the determination of AA. Moreover, introduction of host-guest recognition based on CP6 and AA can bring new properties for enhancing selectivity, sensitivity, and practical application in real environment. Host-guest interactions between CP6 and AA were evaluated by the H-1 NMR spectrum. When compared with other literatures, our method displayed a lower determination limit and broader linear range. To the best of our knowledge, this is the first study carried out for the non-noble transition-metal Co ion, COF, and pillar[6]arene hybrid material in sensing field, which has a potential value in sensing, catalysis, and preparation of advanced multifunction materials.