Tricyclic　antidepressants　(TCAs)　including　amitriptyline　(AT),　doxepin　(DOX)　and　nortriptyline　(NT)　are　the　first-line　drugs　for　the　clinical　treatment　of　depression;　however,　monitoring　TCA　concentrations　in　biological　fluids　and　tissues　is　necessary　to　improve　therapeutic　effect　and　determine　the　cause　of　death　in　patients.　It　is　of　great　significance　to　develop　a　rapid　and　sensitive　method　for　real-time　monitoring　of　TCAs　in　various　biosamples.　In　this　work,　we　fabricated　a　novel　covalent　organic　framework　(COF)　based　solid-phase　microextraction　(SPME)　probe　by　an　in-situ　step-by-step　strategy,　which　was　obtained　by　se-quentially　modifying　1,3,5-tri　(4-aminophenyl)　benzene　(TPB)　and　2,　5-divinylbenzaldehyde　(DVA)　on　the　surface　of　polydopamine　layer.　The　TPB-DVA-COF-SPME　probe　possessed　high　specific　surface　area　(1244　m2　center　dot　g　-　1　),　regular　pores　(3.23　nm),　good　hydrophobicity　and　stability,　resulting　in　efficient　enrichment　for　TCAs.　Furthermore,　the　combination　of　TPB-DVA-COF-SPME　probe　and　ambient　electrospray　ioniza-tion　mass　spectrometry　system　(ESI/MS)　was　firstly　proposed　for　rapid　and　sensitive　determination　of　TCAs　in　biosamples.　As　a　result,　the　developed　method　exhibited　low　limits　of　detection　(LODs)　(0.1-0.5　mu　g　center　dot　L　-　1　),　high　enrichment　factors　(39-218),　and　low　relative　standard　deviations　(RSDs)　for　one　probe　(1.2-3.8%)　and　probe-to-probe　(2.0-3.7%).　Benefiting　from　these　outstanding　performance,　TPB-DVA-COF-SPME　probe　was　further　successfully　applied　to　biosamples　(i.e.,　serum,　liver,　kidney,　and　brain)　with　excellent　reusability,　indicating　the　promising　applicability　of　the　TPB-DVA-COF-SPME-ESI/MS　as　a　pow-erful　tool　for　drug　monitoring.(c)　2023　Elsevier　B.V.　All　rights　reserved.