Sensitive　and　specific　analysis　of　zearalenone　(ZEN)　mycotoxin　in　cereals　for　ensuring　food　safety　is　critical　and　remains　challenging.　Herein,　a　new　gold　nanoparticles　@aptamer-functionalized　hybrid　affinity　monolithic　column　was　proposed　and　employed　for　online　specific　recognition　of　ZEN　by　HPLC.　Characterization　on　the　morphology,　Brunauer–Emmett–Teller　(BET)　surface　area　mechanical　stability　and　specific　performance　of　the　obtained　affinity　monolith　were　investigated.　A　super-high　aptamer　coverage　density　could　reach　3636　pmol/μL,　which　is　preferable　to　gain　an　effective　analysis　of　ZEN　with　high　specificity　and　a　low　interference　of　co-existed　substances　including　typical　α-Zearalenol　(α-ZOL)　and　Aflatoxin　B1　(AFB1).　The　sensitive　recognition　of　trace　ZEN　was　obtained　with　the　limit　of　detection　(LOD)　as　low　as　0.05　ng/mL.　Applied　to　real　cereal　samples,　satisfactory　recoveries　were　obtained　in　the　range　of　91.6　±　1.4%–97.8　±　2.6%　(n　=　3)　in　corn,　93.8　±　3.1%–95.0　±　3.6%　(n　=　3)　in　wheat,　and　90.9　±　4.7%–94.7　±　3.8%　(n　=　3)　in　rice,　respectively.　The　results　on　quantitative　analysis　were　similar　to　that　of　LC–MS　and　better　than　that　obtained　by　using　immunoaffinity　column　(IAC)　or　molecularly　imprinted　polymer　(MIP).　This　protocol　provided　an　efficient　access　to　high-efficient　online　specific　recognition　of　ZEN　in　cereals　by　using　such　an　aptamer-affinity　capillary　monolithic　column.　©　2020