Per-　and　polyfluoroalkyl　substances　(PFASs)　are　potentially　neurotoxic　compounds.　Levels　of　PFASs　in　cerebrospinal　fluid　(CSF)　could　directly　reflect　their　potential　harm　to　the　central　nervous　system.　Because　of　the　variety　of　PFASs　and　the　rarity　of　CSF,　there　is　an　urgent　need　to　establish　a　rapid　online　method　to　detect　a　broad　spectrum　of　PFASs　accurately　and　simultaneously　by　consuming　a　small　amount　of　CSF.　In　this　study,　we　developed　a　fast　and　automated　method　to　analyze　52　PFASs　in　human　CSF　samples　using　online　TurboFlow　ultra-high-performance　liquid　chromatography-tandem　mass　spectrometry.　Our　method　offered　excellent　matrix-matched　standard　curve　linearity　(correlation　coefficient　＞　0.99),　good　limits　of　quantitation　(MLOQs)　(0.01　to　0.08　ng　mL(-1)),　satisfactory　accuracy　(recoveries　of　74.6%-119.1%)　and　precision　(relative　standard　deviations　of　1.4%-13.2%),　small　sample　amount　consumption　(50　mu　L),　and　fast　analysis　time　(18　min　per　sample)　without　complex　sample　pretreatment　procedures.　These　are　advantageous　for　the　high　throughput　screening　of　PFASs　in　environmental　epidemiology　studies.　Repeated　freeze-thaw　experiments　showed　that　it　was　better　to　perform　the　analytical　process　soon　as　possible　after　sample　collection.　The　established　method　was　used　to　analyze　PFASs　in　60　people.　Short-chain　PFASs,　perfluorobutanoic　acid　(PFBA),　perfluoropentanoic　acid　(PFPeA),　and　novel　PFASs　[sodium　2-(N-ethylperfluorooctane-1-sulfonamido)ethyl　phosphate　(SAmPAP),　perfluoroethylcyclohexanesulfonate　(PFECHS),　and　perfluoro-3,　7-dimethyloctanoic　acid　(P37DMOA)]　were　reported　in　CSF　for　the　first　time.　PFBA　and　PFPeA　were　detected　in　all　samples　with　mean　concentrations　of　0.24　and　0.22　ng　mL(-1),　respectively.　We　also　calculated　the　blood-brain　barrier　transmission　efficiency　of　PFASs　(R-PFAS),　and　the　mean　R-PFBA　value　was　above　1,　which　indicated　that　PFBA　might　transfer　from　serum　to　CSF.