Synergistic and Additive Interactions in Essential Oils Obtained from Combined Plant Materials: Enhanced Control of Insect Pests

Essential oils (EOs) from combined plant materials offer a promising alternative to conventional extraction by enhancing chemical diversity and bioactivity. This study evaluated the chemical composition and insecticidal properties of individual and combined plant EOs from <i>Cymbopogon citratus</i>, <i>Eucalyptus camaldulensis</i>, <i>Eucalyptus lehmannii</i>, <i>Salvia rosmarinus</i> and <i>Thymus vulgaris</i> were evaluated against aphids. Binary and ternary combinations were prepared in equal proportions prior to hydrodistillation. GC-MS analysis revealed significant compositional shifts in EOs from combined plant materials. Major compounds in individual oils included citral (53.11%) and neral (29.14%) in <i>C. citratus</i>, thymol (70.84%) in <i>T. vulgaris</i>, and eucalyptol as the predominant compound in <i>E. camaldulensis</i> (66.51%), <i>E. lehmannii</i> (56.99%) and <i>S. rosmarinus</i> (46.56%), respectively. In the combined oils, the relative abundance of these constituents was altered, and in some cases new constituents were introduced. Principal Component Analysis (PCA) and Hierarchical Cluster Analysis (HCA) revealed that combined plant EOs clustered near their parental oils, indicating compositional inheritance. Contact toxicity assay against <i>Aphis fabae</i> demonstrated enhanced efficacy of the combined oils, with reduced LC₅₀ values (1.39 µL mL^-1 for <i>E. camaldulensis</i> + <i>T. vulgaris</i>) and synergistic interactions, indicated by a co-toxicity coefficient (CTC) of 221.58 and elevated synergistic factors. Pearson correlation analysis and Partial Least Squares (PLS) regression jointly identified Acorenone B and thymol as negatively, and caryophyllene as positively correlated compounds, all with relatively high contribution to insecticidal activity, ranking highest with a Variable Importance in Projection (VIP) scores > 1.0. While PLS model had modest predictive power, the integration of these statistical approaches supports the insecticidal potential of combined plant-derived EOS in laboratory bioassays and indicates their relevance to sustainable crop protection.