In the present study,the potential anti-allergic constituents in Corni Fructus were investigated. The anti-allergic components of Corni Fructus were preliminarily screened by hyaluronidase inhibition assay. The fractions with high hyaluronidase inhibition activity were further evaluated by the degranulation inhibition test of RBL-2H3 cells in vitro. The anti-allergic components were analyzed by gas chromatography-mass spectrometry (GC-MS). The results showed that the petroleum ether extract of Corni Fructus (COPE) had a significantly anti-hyaluronidase activity with an IC₅₀ value of 3.11 mg/mL. Four fractions with higher anti-hyaluronidase activity were separated by silica gel column chromatography,among which COPE-8 had a more significant inhibitory effect on RBL-2H3 cell degranulation in a dose-dependent manner and showed low cytotoxicity. After treatment with 20 μg/mL COPE-8,the histamine and β-hexosaminidase released from RBL-2H3 cells were reduced by 46% and 39%,respectively. The main active components in COPE-8 were α-amyrin,β-amyrin,and sclareol,by GC-MS analysis. These findings signpost that COPE-8 has remarkable anti-allergic activity and has potential application in the development of anti-allergic active substances.