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We elucidated the changes in flavonoids (icariin, epimedin A, epimedin B, and epimedin C) in E. sagittatum leaves. The sum of primary flavonoids content reached a maximum (11.74%) at 20 days following the high-concentration Fe2+ (2500 mg·L-1) treatment. We examined the FT-IR spectra characteristics of E. sagittatum leaf samples utilising the FT-IR strategy, and constructed an OPLS-DA model and identified characteristic peaks to obtain classified recognition of E. sagittatum. Further, wid