Evidence of the Antioxidant and Anti‐Inflammatory Potential of Chemically Characterized <i>Cladanthus mixtus</i> Flower Essential Oils: Insights From In Vitro, In Vivo, and In Silico

Abstract

This study investigates the chemical composition, antioxidant, and anti-inflammatory effects of Cladanthus mixtus (L.) Chevall. flower essential oil (CMEO). Gas Chromatography-Mass Spectrometry (GC-MS) analysis allowed for the identification of 29 compounds representing 92.73% of the total oil. The major constituent was (E)-2-Methylbut-2-en-1-yl methacrylate (32.62. Antioxidant activity yielded IC50 values of 566 ±7 µg/mL for DPPH, 491 ± 10 µg/mL for ABTS, and an EC50 of 235 ± 12 µg/mL for FRAP. These results demonstrate significant radical scavenging and reducing potentials compared to BHT (147 ±2 µg /mL for DPPH) and ascorbic acid (201 ± 5 µg/mL for ABTS). The anti-inflammatory effects of CMEO were assessed through in vitro models, revealing IC50 values for BSA denaturation, heat-induced, and hypotonicity-induced hemolysis tests at 508.6 ± 11, 456.4 ± 9, and 589 ±7 µg/mL, respectively. These values were compared against standards such as Diclofenac sodium (IC50 = 265.6 ±7 µg/mL) and Indomethacin (IC50 = 121.3 ±6 µg/mL). Molecular docking studies provided further insights into the bioactivity of β-Caryophyllene, humulene, and caryophyllene oxide, which showed strong interactions with inflammatory mediators, including TNF-α, COX1, COX2, and 5-LOX enzymes. These findings highlight CMEO as a potential source of natural antioxidant and anti-inflammatory compounds.