The structural properties and nonbonding interactions of a menthol-based deep eutectic solvent (DES) were investigated in detail employing experimental and computational methods. A mass spectrometry analysis confirmed the formation of 1:1 l-menthol/acetic acid. A molecular dynamics simulation was used to figure out energetically most favorable cluster conformers of the 1:1 l-menthol/acetic acid system. Density functional theory at the ωB97XD/6-311G (d,p) level of theory was employed to optimize the isolated structures and to calculate their thermochemical properties. Both experimental and computed IR spectra were analyzed for the samples. Additionally, vibrational circular dichroism (VCD) spectra of the samples were measured to prove the chirality transfer. Principal component analysis (PCA) was used to make the data interpretation more vivid. All the spectral data analyses and nanostructure elucidation proved the spontaneous formation of the DES through the formation of strong hydrogen bonding. Experimental solvatochromism and computed highest occupied molecular orbital-lowest unoccupied molecular orbital gaps validated the reasoning. Moreover, comparative VCD and IR spectral analyses clearly indicated a chirality transfer from the chiral menthol to achiral acetic acid. This study suggests that various techniques, such as mass spectrometry, IR, solvatochromism, and computed IR-VCD could be useful and important tools to elucidate nanostructure and nonbonding interactions of a DES. VCD could be used as an excellent complementary technique to IR spectroscopy for a chiral molecule-based DESs.