Withania somnifera (ashwagandha) root extract is a popular health supplement, with purported health benefits including prevention and treatment of various diseases, i.e. anxiety, stress, etc. The objective of this experiment was to evaluate the impact of Energy of Consciousness Healing Treatment (The Trivedi Effect®) on the characteristic properties of the phytoconstituents present in 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 another 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 liquid chromatographic data of the control and Biofield Energy Treated samples revealed that the retention time of the 25 chromatographic peaks remained same, whereas the peak area% i.e. the relative amount of the phytoconstituents was altered significantly. The peak area% of the treated ashwagandha root extract representing the phytoconstituents was significantly decreased in the range of 6.02% to 39.74% at Rt of 5.2, 5.3, 5.4, 5.6, 5.7, 6.9, 7.1, 7.3, 7.8, 7.9, 8.0, and 8.5 minutes compared to the control sample. On the contrary, the peak area% of the other phytoconstituents present in the treated sample was significantly increased in the range of 4.12% to 82.32% at Rt of 5.1, 6.4, 6.6, 6.8, 8.1, 8.2, 8.4, 8.6, 8.8, 8.9, 9.0, 9.1, and 9.2 minutes, respectively compared to the control sample. A total of 16 withanolides such as sitoindoside IX, viscosa lactone B, dihydrowithanolide D, withanolide A, withaferin A, ixocarpalactone A, withanolide sulfoxide, etc. were proposed from the molecular mass at m/z 605, 489, 473, 471, 505, and 992 at the retention times of 6.9, 7.1, 7.8, 8.2, 8.5, and 9.2 minutes with the help of LC-MS, GC-MS and NMR data of both the samples. Subsequently, the mass peak intensities of the treated sample were significantly changed in the range of -61.24% to 106.61% compared with the control sample at the same retention time. These findings suggest that Energy of Consciousness Healing Treatment could be advantageous 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 ashwagandha extract that might provide better therapeutic response against inflammatory diseases, immunological disorders, arthritis, stress, cancer, diabetes, sexual disorders, aging, and other chronic infections.