To investigate whether McLTPⅡ.9 (the non-specific lipid transfer protein from Mentha canadensis Linn.) possesses affinity for monoterpenoids, protein structural analysis was performed on signal peptide-truncated McLTPⅡ.9 (McLTPⅡ.9-ΔSP), and six monoterpenoids, namely menthol, menthone, linalool, pulegone, limonene, and α-pinene were selected based on previous research findings for molecular docking prediction, followed by in vitro binding activity validation. The results show that McLTPⅡ.9-ΔSP harbors a core nsLTPⅡ domain, its tertiary structure is mainly composed of α-helices and loop regions, moreover, the protein structure of McLTPⅡ.9-ΔSP is reasonable for further analysis. The molecular docking prediction results indicate that McLTPⅡ.9-ΔSP exhibits binding interactions with all six monoterpenoids, among which the affinity with linalool and α-pinene is particularly prominent, and Leu8, Leu45, Phe48, and Ile49 may serve as important amino acid binding sites. The in vitro binding activity experiment results show that McLTPⅡ.9-ΔSP could bind to all the aforementioned monoterpenoids, and the binding capacity ranking (from high to low) is generally consistent with the molecular docking affinity prediction result, in the following order: linalool, menthol, α-pinene, menthone, limonene, pulegone. The comprehensive analysis indicates that McLTPⅡ.9 is capable of binding monoterpenoids and possesses potential transport functions