The aim of this work was to determine phenolic profile and antioxidant properties of methanolic (MEP) and ethanolic (EEP) extracts of monofloral bee-collected sunflower pollen from Serbia. In total, 37 different polyphenols (19 flavonol glycosides and 18 other polyphenols) were identified by UHPLC/MS-MS Orbitrap analysis. The different phenolic profile of MEP and EEP was observed with flavonols as most abundant, 94.1% (MEP) and 93.4% (EEP), out of which flavonoid glycosides represented 73.2% (MEP) and 66.1% (EEP). The higher amount of total phenolics was recorded in EEP (200.58 mg/kg DW) than in MEP (244.44 mg/kg DW). The quercetin 3-O-galactoside was the major compound in both extracts, 112.86 mg/kg in MEP and 128.64 mg/kg in EEP. The presence of coumaroylated and acetylated flavonoid glycosides were registered, among which the quercetin 3-O-(6“-acetyl)hexoside was recored in the significant amount in MEP (111.10 mg Q3G/kg DW), whereas in EEP the content of coumaroylated kaempferol glycosides (137.95 mg Q3G/kg DW) was significantly higher. The ferric reducing power and ABTS radical scavenging activity assays revealed that pollen extracts possessed significant antioxidant activity. The monofloral bee-collected pollen of Helianthus annuus L. can be used as good food supplement with high flavonoids content and antioxidant activity. Display omitted •Polyphenolic profile of sunflower bee-collected pollen is determined.•In total, 37 different phenolic compounds were identified/quantified.•The different phenolic profile of methanolic and ethanolic extracts was observed.•Pollen extracts possessed considerable antioxidant activity.•Due to significant flavonoids content pollen is potentially good food supplement.