Withania somnifera (Ashwagandha) root extract is very useful herbal medicine since from ancient time. The current study designed to investigate the impact of The Trivedi Effect® - Energy of Consciousness Healing Treatment on the structural properties of the ashwagandha root hydroalcoholic extract using LC-MS, GC-MS, and NMR spectroscopy. Ashwagandha root extract was divided into two parts – one part was control (without treatment), while other part was treated with the Energy of Consciousness Healing Treatment remotely by seven renowned Biofield Energy Healers and defined as the Biofield Energy Treated sample. The LC-MS analysis revealed that the retention time of the phytoconstituents remained same in both the control and treated samples, whereas the peak area % at respective retention time was significantly altered. The peak area% of the treated sample at Rt of 5.3, 5.5, 6.4, 6.5, 6.8, 6.9, 7.1, 7.3, 7.8, 7.9, 8.0, 8.2, 8.4, 8.6, 8.8, 9.0, 9.1, and 10.1 minutes were significantly reduced by 0.57% to 38.10% compared to the control sample. In addition, the peak area% of the treated sample at Rt of 5.7, 6.7, and 8.1 minutes were significantly increased by 16.00%, 244.44% and 19.62%, respectively compared with the control sample. A total of 21 withanolides such as withanoside IV, coagulin Q, viscosa lactone B, withanolide A, withaferin A, withanone, withanolide D, ixocarpalactone A and withanolide sulfoxide, ixocarpalactone A, withanolide sulfoxide, withanolide B, etc. were proposed with their structure from the molecular mass at m/z 783, 569, 621, 489, 473, 767, 471, 505, 992, and 455 at retention times of 6.4, 6.5, 6.8, 7.1, 7.9, 8.1, 8.4, 9.1, and 10.1 minutes with the help of GC-MS and NMR data of both the control and Biofield Energy Treated samples. There were significant changes observed in the peak intensity values at the same retention time in the Biofield Energy Treated sample in the range of -75.32% to 108.51% compared with the control sample. These findings suggest that The Trivedi Effect® - Energy of Consciousness Healing Treatment could be beneficial for altering the concentration of the phytoconstituents in the ashwagandha root extract by modifying their intrinsic physicochemical properties, which might be helpful to improve the bioavailability of active constituents of W. somnifera extract that might provide better therapeutic response against inflammatory diseases, immunological disorders, stress, arthritis, cancer, diabetes, sexual disorders, aging and other chronic infections.