Solid-phase　microextraction　(SPME)　coupled　with　electrospray　ionization　mass　spectrometry　(ESI-MS)　was　developed　for　rapid　and　sensitive　determination　of　endogenous　androgens.　The　SPME　probe　is　coated　with　covalent　organic　frameworks　(COFs)　synthesized　by　reacting　1,3,5-tri(4-aminophenyl)benzene　(TPB)　with　2,5-dioctyloxybenzaldehyde　(C8PDA).　This　COFs-SPME　probe　offers　several　advantages,　including　enhanced　extraction　efficiency　and　stability.　The　analytical　method　exhibited　wide　linearity　(0.1–100.0 µg　L−1),　low　limits　of　detection　(0.03–0.07 µg　L−1),　high　enrichment　factors　(37–154),　and　satisfactory　relative　standard　deviations　(RSDs)　for　both　within　one　probe　(4.0–14.8%)　and　between　different　probes　(3.4–12.7%).　These　remarkable　performance　characteristics　highlight　the　reliability　and　precision　of　the　COFs-SPME-ESI-MS　method.　The　developed　method　was　successfully　applied　to　detect　five　kinds　of　endogenous　androgens　in　female　serum　samples,　indicating　that　the　developed　analytical　method　has　great　potential　for　application　in　preliminary　clinical　diagnosis.　Graphical　abstract:　(Figure　presented.).　©　The　Author(s),　under　exclusive　licence　to　Springer-Verlag　GmbH　Austria,　part　of　Springer　Nature　2024.