Time-consuming　or　tedious　operation　in　multiple-step　process　might　is　the　obstacle　for　efficiently　preparing　aptamer-affinity　monolithic　column.　Here,　a　new　and　facile　strategy　to　prepare　aptamer-based　hybrid　affinity　monolith　in　＇one-pot＇　at　room　temperature　was　exploited,　in　which　UV　light-initiated　free-radical　polymerization　and　＇thiol-ene＇　click　reaction　were　implemented　simultaneously.　Only　7　min　was　cost　for　finishing　the　polymerization　reaction,　which　was　only　1/100　of　that　for　the　traditional　thermal　polymerization.　Using　ochratoxin　A　(OTA)　as　the　model　analyte,　the　recipe　for　photo-initiated　polymerization　was　optimized,　and　SEM　morphology,　FTIR,　EDS,　pore　size　distribution　and　specific　recognition　performance　were　also　studied.　Compared　with　traditional　thermal　polymerization,　the　resultant　monolith　was　achieved　more　facilely　and　displayed　better　results　such　as　more　homogeneous　skeleton　structure,　higher　reaction　efficiency　of　aptamer　(＞88.2%)　and　better　specific　selectivity　to　OTA.　Besides,　an　extremely　low　nonspecific　adsorption　of　analogues　was　obtained　and　showed　a　level　at　only　1/25　of　that　in　the　similar　aptamer-affinity　monolith　prepared　by　thermal　polymerization.　Applied　to　beer　and　red　wine　samples,　good　recovery　yields　about　99.7　±　4.0%　–101.2　±　2.3%　(n　=　3)was　achieved　with　the　acceptable　RSDs.　It　would　open　up　a　rapid　and　promising　access　to　efficiently　preparing　high-performance　aptamer-based　affinity　monolithic　columns　for　online　specific　recognition.　©　2021　Elsevier　B.V.