Tilia　tuan　honey　has　many　biological　activities,　and　the　source　of　biological　activities　is　inseparable　from　the　protein　and　peptides　in　tilia　tuan　honey.　Antioxidant　activity　is　one　of　the　many　biological　activities　of　tilia　tuan　honey.　In　this　paper,　the　composition　and　structure　of　trypsinolysis　peptides　in　tilia　tuan　honey　were　identified　by　ultra-high　performance　liquid　chromatography-quadrupole/electrostatic　field　orbitrap　high　resolution　mass　spectrometry　(UPLC-Q-Exactive),　and　the　relationship　between　trypsinolysis　peptides　in　tilia　tuan　honey　and　antioxidant　activity　was　discussed.　Tilia　tuan　honey　protein　was　precipitated　by　20%trichloroacetic　acid　and　lyophilized.　The　lyophilized　protein　was　dissolved　in　100　mmol/L　NH4HCO3　buffer　solution.　Dithiothreotol　was　added　to　reduce　protein,　iodoacetamide　was　added　to　alkylate　protein,　and　500　mmol/L　dithiothreotol　solution　was　added　to　the　final　protein　solution　to　stop　the　alkylation　reaction,　then　the　protein　was　hydrolyzed　by　trypsin.　The　enzymatic　peptides　was　enriched　and　purified　by　C18　solid　phase　extraction　column.　Gradient　elution　was　carried　out　with　0.1%　formic　acid　(A)　and　acetonitrile　(B)　as　mobile　phase.　The　enzymatic　hydrolyzed　tilia　tuan　honey　peptides　was　identified　by　Full　MS/ddMS2　mode　of　high　resolution　mass　spectrometry　(Q-Exactive).　The　results　of　mass　spectrometry　were　analyzed　by　MaxQuant　software,　and　compared　on　Uniprot　by　Blast.　A　total　of　52　peptides　in　tilia　tuan　honey　were　identified,　71.15%　of　which　were　derived　from　major　royal　jelly　proteins　(MRJPs),　and　the　identified　peptides　belonged　to　10　proteins,　of　which　6　(60%)　were　MRJP　family　proteins.　Reproducibility　analysis　of　tilia　tuan　honey　peptides　mass　spectrometry　results,　more　than　two　repetitive　mass　spectrometry　can　achieve　reliable　identification　of　tilia　tuan　honey　peptides　(87.1%).　According　to　the　structure-activity　relationship　of　honey　peptides　reported　in　the　literature,　11　peptides　with　antioxidant　activity　were　screened　out.　Four　different　antioxidant　activities　were　tested　after　chemical　synthesis　of　11　peptides.　The　results　showed　that　the　antioxidant　activities　of　these　11　peptides　were　significantly　different,　and　1　g/L　VIYEWK　showed　strong　2,2＇-azinobis-(3-ethylbenzthiazoline-6-sulphonate)　(ABTS)　free　radical　scavenging　activity　(33.8±0.6%),　2,2-diphenyl-1-picryhydrazyl　(DPPH)　free　radical　scavenging　activity　(96.5±0.0%),　and　showed　a　certain　reducing　power(0.010±0.001)　and　Fe2+　chelating　activity　(4.0±0.2%).　The　method　can　quickly　and　effectively　identify　the　structure　of　tilia　tuan　honey　peptides,　provide　a　basis　for　revealing　the　relationship　between　tilia　tuan　honey　peptides　structure　and　antioxidant　activity.　But　the　results　of　antioxidant　activity　in　vitro　can　not　fully　explain　the　antioxidant　activity　of　peptides.　Therefore,　it　is　necessary　to　study　the　antioxidant　activity　of　peptides　in　vivo　and　the　antioxidant　activity　of　peptides　is　the　result　of　the　interaction　of　various　amino　acids.　The　modification　of　amino　acids　of　antioxidant　peptides　can　be　considered　to　determine　the　specific　amino　acids　that　play　an　antioxidant　role.　©　2020,　Editorial　Board　of　Journal　of　Chinese　Mass　Spectrometry　Society.　All　right　reserved.