Imidazole derivatives have attracted significant interests in recent time for their usefulness in synthetic heterocyclic chemistry, analytical chemistry and pharmacology. Aim of present study was to evaluate the impact of biofield treatment on two imidazole derivatives (i.e., imidazole and 2-methylimidazole) by various analytical methods. The biofield treatment was done by Mr. Trivedi on both the compounds and both control and treated samples of imidazole and 2-methylimidazole were characterized with respect to physical, and structural properties using X-ray diffraction (XRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), Fourier transform infrared (FT-IR), ultraviolet-visible (UV-Vis) spectroscopy, and Gas chromatography-Mass spectrometry (GC-MS). X-ray diffraction study revealed that crystallite size varied in a different way for imidazole and 2-methylimidazole due to the presence of methyl group in 2-c position although their core was same. Treated sample of imidazole showed a slight increase in crystallite size (6.5%); however, treated 2-methylimidazole showed a significant increase (166.68%) in crystallite size along with decrease in peak intensity as compared to control. The latent heat of fusion (ΔH) of imidazole was increased up to 0.62% in treated sample as compared to control; whereas in treated 2-methylimidazole, the ΔH was decreased by 22% as compared to control. Maximum degradation temperature (Tmax) from TGA of imidazole was remained same but 2-methylimidazole was increased by 1.5% as compared to control. FT-IR spectra showed slight change in stretching frequencies of treated imidazole and 2-methylimidazole as compared to control. Both the imidazole and 2-methylimidazole showed similar UV absorbance maxima as compared to respective control sample. GC-MS data revealed that isotopic abundance ratio of either 13C/12C or 15N/14N or 2H/1H (PM+1/PM) of treated imidazole was significantly increased up to 232.51% as compared to control, however, isotopic abundance ratio of 13C/12C or 15N/14N or 2H/1H (PM+1/PM) of treated 2-methylimidazole showed a minor change from -1.68 upto 1.68% as compared to control. Overall, the experimental results suggest that biofield treatment has significant effect on structural and thermal properties of imidazole and 2-methylimidazole.