O-Toluic acid (OTA) and m-toluic acid (MTA) are two isomers of toluic acid that act as an important organic intermediates, mostly used in medicines and pesticides. The aim of the study was to evaluate the impact of biofield energy treatment on isotopic abundance ratios of 2H/1H, 13C/12C, (PM+1)/PM and 18O/16O, (PM+2)/PM, in toluic acid isomers using gas chromatography-mass spectrometry (GC-MS). The OTA and MTA samples were divided into two parts: control and treated. The control sample remained as untreated, while the treated sample was further divided into four groups as T1, T2, T3, and T4. The treated group was subjected to biofield energy treatment. The GC-MS spectra of both the isomers showed five m/z peaks due to the molecular ion peak and fragmented peaks of toluic acid derivatives. The isotopic abundance ratio of (PM+1)/PM and (PM+2)/PM were calculated for both the isomers and found significant alteration in the treated isomers. The isotopic abundance ratio of (PM+1)/PM in treated samples of OTA was decreased and then slightly increased upto 2.37% in T2, where the (PM+2)/PM in treated OTA, significantly decreased by 55.3% in T3 sample. Similarly, in case of MTA, the isotopic abundance ratio of (PM+1)/PM in the treated sample showed a slight increase the (PM+2)/PM was decreased by 11.95% in T2 as compared to their respective control. GC-MS data suggests that the biofield energy treatment on toluic acid isomers had significantly altered the isotopic abundance of 2H, 13C, and 18O in OTA and MTA as compared to the control.