Ofloxacin is a broad-spectrum antibiotic, which is useful for the treatment of many Gram-positive and Gram-negative bacterial infections. This study was designed to investigate the impact of The Trivedi Effect®-Biofield Energy Healing Treatment on the structural properties and the isotopic abundance ratio of ofloxacin using LC-MS and GC-MS spectroscopy. Ofloxacin sample was divided into two parts, one part of ofloxacin was considered as control (no Biofield Energy Treatment was provided), while the second part was treated with The Trivedi Effect®-Consciousness Energy Healing Treatment remotely by a renowned Biofield Energy Healer, Alice Branton and termed as a treated sample. The LC-ESI-MS spectra of both the samples of ofloxacin at the retention time 3.02 minutes exhibited the mass of the protonated molecular ion peak at m/z 362.17 [M+H]+ (calculated for C18H21FN3O4+, 362.15). The LC-MS based isotopic abundance ratio of PM+1/PM in the treated ofloxacin was significantly increased by 28.65% compared with the control sample. Thus, 13C, 2H, 15N, and 17O contributions from (C18H21FN3O4)+ to m/z 363.17 in the treated sample were significantly increased compared with the control sample. In the GC-MS the retention times of the treated sample (23.08 minute) was found to be close to those of the control sample (22.87 minutes). The peak area % of treated ofloxacin was increased by 0.7% compared to the control sample. Similarly, the GC-MS based isotopic abundance ratio of PM+1/PM in the treated ofloxacin was increased by 3.73% compared with the control sample. Hence, 13C, 2H, 15N, and 17O contributions from (C18H21FN3O4)+ to m/z 318 in the treated sample were significantly increased compared with the control sample. On the contrary, the isotopic abundance ratio of PM+2/PM in the treated ofloxacin was decreased by 10.60% compared with the control sample. Therefore, 18O contributions from (C18H21FN3O4)+ to m/z 319 in the treated sample were significantly decreased compared with the control sample. The LC-MS and GC-MS based isotopic abundance ratios of PM+1/PM (2H/1H or 13C/12C or 15N/14N or 17O/16O) and PM+2/PM (18O/16O) and peak area % in the treated ofloxacin were significantly altered compared to the control sample. The new form of treated ofloxacin would be better soluble and bioavailable compared to the control sample and would be very useful to design better pharmaceutical formulations that might offer better therapeutic response against urinary tract infections, infectious diarrhoea, infections of the urethra and cervix, pneumonia, cellulitis, chronic bronchitis, tuberculosis, prostatitis, plague, other bacterial infections, etc. Journal Web URL:- https://crimsonpublishers.com/sbb/fulltext/SBB.000586.php