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Waldheimia glabra is traditionally used as incense and as an anti-influenza drug by Tibetans in China. Here, we collected W. glabra from the Gangs Rinpoche mountain at an altitude of 5200 m, and analyzed its essential oil by gas chromatography-mass spectrometry (GC-MS) combined with the retention indices (RI). Twenty-seven compounds, representing 72.4% of the total essential oil, were identified, including α-bisabolol (20.2%), valeranone (11.8%), chamazulene (9.9%), spathulenol (8.2%), β-caryophyllene (6.1%), and caryophyllene oxide (5.2%). Bioactivity evaluation of the essential oil revealed that it exhibited potent anti-influenza effect on viruses H3N2 and anti-inflammatory effect by inhibiting the lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 macrophages, but no anti-complementary activity.