Withania somnifera root extract which contains withanolides as major active constituent and it is used traditionally for the prevention and treatment of several diseases. The objective of the current study was to investigate the impact of The Trivedi Effect® - Consciousness Energy Healing Treatment on the structural properties of the ashwagandha root 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 Consciousness Energy Healing Treatment remotely by twenty 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 the control and Biofield Energy Treated samples, whereas the peak area% i.e. the relative amount of the phytoconstituents at respective retention time was significantly altered. The peak area% at Rt of 5.6, 6.8, 6.9, 7.2, 7.9, 8.4, 8.5, 8.6, and 9.2 minutes of the treated sample were increased significantly in the range of 1.46% to 253.06% compared to the control sample. In the contrary, the peak area% of the treated sample at Rt of 6.4 and 8.2 minutes were significantly decreased by 12.72% and 17.35%, respectively with respect to the control sample. A total of 16 withanolides such as withanoside IV, coagulin Q, viscosa lactone B, dihydrowithanolide D, withanolide A, withaferin A, withanone, withanolide D, ixocarpalactone A, withanolide sulfoxide, withanolide B, etc. were proposed with their structure from the molecular mass at m/z 783, 621, 489, 473, 767, 471, 505, and 992 at retention times of 6.4, 6.8, 7.2, 7.9, 8.2, 8.4, and 9.1 minutes, respectively with the help of LC-MS, GC-MS and NMR data of both the control and Biofield Energy Treated samples. The mass peak intensities of the Biofield Energy Treated sample were significantly changed in the range of -81.36% to 1720.90% compared with the control sample at the same retention time. These findings suggest that The Trivedi Effect® - Consciousness Energy 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.