A　photo-initiated　polymerized　oligonucleotide-grafted　hydrophilic　affinity　monolithic　column　was　synthesized　in　situ,　and　exploited　for　selective　in-tube　solid　phase　micro-extraction　(IT-SPME)　protocol　towards　the　sensitive　detection　of　ochratoxin　A　(OTA).　Only　7　min　was　required　for　the　rapid　polymerization　of　aptamer-based　affinity　monolith,　which　was　much　less　than　the　reaction　time　of　most　thermal　polymerization　(12-16　h)　and　sol-gel　chemistry　methods　(up　to　52　h).　Characterizations　such　as　polymerization　recipes,　structure　morphology,　FTIR　spectrum,　elemental　mapping,　mechanical　stability,　and　specific　recognition　performance　were　evaluated.　A　significantly　hydrophilic　nature　with　a　low　contact　angle　of　15　degrees　was　observed,　and　a　mixed-mode　mechanism　including　aptamer　affinity　recognition　and　hydrophilic　interaction　(HI)　was　employed.　By　coupling　with　HPLC-fluorescence　detection,　the　highly　specific　online　recognition　performance　was　achieved　with　an　extremely　low　nonspecific　adsorption　of　the　analogues.　The　calibration　curve　of　OTA　was　obtained　in　the　concentration　range　0.05-50.00　ng.mL(-1)　with　a　limit　of　detection　(LOD,　S/N　=　3)　of　0.012　ng.mL(-1).　Applied　to　sample　analysis,　acceptable　recovery　yields　of　95.1　+/-　1.4%　-　99.5　+/-　2.2%　(n　=　3)　were　obtained　in　beer　and　red　wine.　The　proposed　method　lighted　a　promising　way　to　efficiently　preparing　a　hydrophilic　aptamer-affinity　monolith　for　highly　specific　recognition　of　trace　mycotoxin　by　IT-SPME　coupled　with　HPLC.