Comprehensive Phytochemical Profiling of Iris songarica Rhizomes and Evaluation of Their Anti-Inflammatory Activity In Vivo

The chemical composition of Iris songarica rhizome extracts was systematically investigated using GC-MS and UHPLC-MS. Their biological activity was further evaluated in vivo. The chloroform rhizome extract contained 33 identified compounds distributed across five main classes. Flavonoids predominated (50.7% of total ionic current), with tectochrysine (42.15%) as the major component, followed by 3,7-dihydroxy-2-(3,4-dimethoxyphenyl)-4H-chromene-4-one (5.18%) and a naringenin derivative (3.99%). Fatty acid esters comprised 30.6%, dominated by linoleic acid ethyl ester (11.05%), ethyl oleate, and hexadecanoic acid ethyl ester. Phenolic and aromatic compounds accounted for 14.24%, including (E)-4-(3-hydroxyprop-1-en-1-yl)-2-methoxyphenol and flamenol. Quantitative HPLC revealed hesperetin (69.72 µg/mL) and fisetin (12.32 µg/mL) as predominant in the 50% aqueous ethanol extract, and cinarin (6.28 µg/mL) in the ethyl acetate root extract. HPLC-MS identified 25 polyphenols, mainly isoflavonoids and flavones, with key markers songaricol, irilin B, tectorigenin, irisflavone A, and irizon B, some reported for the first time in Kazakhstan irises. Biological evaluation demonstrated potent activity: the 50% aqueous ethanol extract inhibited xylene-induced ear oedema in mice by 72.7% at 300 mg/kg, comparable to diclofenac (90.9%), without observable toxicity. These findings confirm I. songarica as a valuable source of bioactive polyphenols with anti-inflammatory potential.