Cholecalciferol is a fat-soluble vitamin (Vitamin D3). It is used for the prevention and treatment of rickets, osteoporosis, arthritis, Parkinson's and Alzheimer's diseases, etc. In this study, the LC-MS and GC-MS analytical methods were used to characterize and evaluate the isotopic abundance ratios of PM+1/PM and PM+2/PM of the Trivedi Effect®-Consciousness Energy Healing Treated cholecalciferol compared to the control sample. The test sample cholecalciferol was divided into control and treated parts. Only, the treated cholecalciferol was received the Trivedi Effect®-Consciousness Energy Healing Treatment remotely by a renowned Biofield Energy Healer, Mahendra Kumar Trivedi. The LC-MS spectra of both the samples at retention time (Rt) ~22 min exhibited the mass of the molecular ion peak at m/z 385.25 (called for C27H45O+, 385.35). The LC-MS based isotopic abundance ratio of PM+1/PM in the treated cholecalciferol was increased by 2.93% compared with the control sample. Similarly, the GC-MS based isotopic abundance ratio of PM+1/PM and PM+2/PM in the treated cholecalciferol was increased by 1.53% and 3.22%, respectively compared with the control sample. Hence, 13C, 2H, 17O and 18O contributions from C27H44O+ to m/z 386 and 387 in the treated cholecalciferol were significantly increased compared with the control sample. The isotopic abundance ratios of PM+1/PM (2H/1H or 13C/12C or 17O/16O) and PM+2/PM (18O/16O) in the treated cholecalciferol were significantly increased compared to the control sample. The increased isotopic composition of the Trivedi Effect®-Consciousness Energy Healing Treated cholecalciferol might have altered the neutron to proton ratio in the nucleus via the possible mediation of neutrino. The increased isotopic abundance ratio of the treated cholecalciferol may increase the intra-atomic bond strength, stability, and shelf-life. The new form of cholecalciferol would be more efficacious pharmaceutical/nutraceutical formulations for the prevention and treatment of various diseases such as vitamin D deficiency, rickets, osteoporosis, arthritis, multiple sclerosis, cancer, mental disorders, diabetes mellitus, cardiovascular diseases, hypertension, infections, influenza, cognitive impairment in older adults, Parkinson's and Alzheimer's diseases, dementia, autoimmune disease, glucose intolerance, multiple sclerosis, etc. Journal Web URL:- https://www.scitcentral.com/article.php?journal=39&article=2123&article_title=Biofield%20Energy%20Treatment%20and%20its%20Impact%20on%20the%20Isotopic%20Abundance%20of%20Cholecalciferol%20(Vitamin%20D3)