A　novel　mercaptotetrazole-silica　hybrid　monolithic　column　was　prepared　for　capillary　liquid　chromatography,　in　which　the　thiol-end　mercaptotetrazole　was　mixed　with　hydrolyzed　γ-methacryloxypropyltrimethoxysilane　and　tetramethyloxysilane　for　the　co-polycondensation　and　thiol-ene　click　reaction　in　a　one-pot　process.　The　effects　of　the　molar　ratio　of　silanes,　the　amount　of　mercaptotetrazole,　and　the　volume　of　porogen　on　the　morphology,　permeability　and　pore　properties　of　the　as-prepared　mercaptotetrazole-silica　hybrid　monoliths　were　investigated　in　detail.　A　series　of　test　compounds　including　alkylbenzenes,　amides　and　anilines　were　employed　for　evaluating　the　retention　behaviors　of　the　mercaptotetrazole-silica　hybrid　monolithic　columns.　The　results　demonstrated　that　the　mercaptotetrazole-silica　hybrid　monoliths　exhibited　hydrophobic,　hydrophilic　as　well　as　ion-exchange　interaction.　The　run-to-run,　column-to-column　and　batch-to-batch　reproducibilities　of　the　mercaptotetrazole-silica　hybrid　monoliths　were　satisfactory　with　the　relative　standard　deviations　less　than　1.4　(n = 5),　3.9　(n = 3)　and　4.0%　(n = 5),　respectively.　In　addition,　the　mercaptotetrazole-silica　hybrid　monolith　was　further　applied　to　the　separation　of　sulfonamides,　nucleobases　and　protein　tryptic　digests.　These　successful　applications　confirmed　the　promising　potential　of　the　mercaptotetrazole-silica　hybrid　monolith　in　the　separation　of　complex　samples.　©　2017　WILEY-VCH　Verlag　GmbH　&　Co.　KGaA,　Weinheim