A　poly　(methacrylic　acid-co-divinyl-benzene)　[poly　(MAA-co-DVB)]　monolithic　column　was　specially　prepared　according　to　the　chemical　structures　of　the　three　mycotoxins　of　aflatoxin　B1,　zearalenone　and　sterigmatocystin,　and　used　for　in-tube　solid-phase　microextraction　(in-tube　SPME)　of　the　selected　mycotoxins.　The　poly　(MAA-co-DVB)　monolithic　column　was　characterized　in　detail,　and　exhibited　effective　hydrophobic,　pi-pi,　and　hydrogen　bonding　interactions　towards　the　target　analytes.　By　coupling　the　poly　(MAA-co-DVB)　monolithic　column-based　in-tube　SPME　with　high-performance　liquid　chromatography,　a　simple,　sensitive　and　matrix　effect-free　method　for　determination　of　mycotoxins　was　developed.　Some　important　parameters　related　to　extraction　efficiency　were　optimized　systematically.　Under　the　optimal　conditions,　the　enrichment　factors　for　the　three　mycotoxins　ranged　from　71.9　to　98.7.　Recoveries　of　mycotoxins　in　rice　grains　with　three　spiked　concentration　levels　were　in　the　range　of　78.0-102.8%,　with　satisfactory　relative　standard　deviations　(n　=　4)　of　2.96-4.74%.　Good　linearity　was　obtained　with　correlation　coefficients　larger　than　0.99,　and　the　detection　limits　(S/N　=　3)　of　the　three　mycotoxins　were　between　0.69　and　2.03　mu　g　kg(-1).　The　developed　poly　(MAA-co-DVB)　monolithic　column　based　in-tube　SPME-HPLC　method　was　successfully　used　to　sensitively　determine　trace　mycotoxins　in　practical　samples　of　rice　grain　without　interference　peaks,　indicating　that　it　is　promising　for　the　detection　of　trace　mycotoxins　in　complex　grain　samples.