Ashwagandha (Withania somnifera) extract is a popular health supplement, used in many nutraceutical and pharmaceutical formulations for good health benefits along with the prevention and treatment of various human diseases. The purpose of this study was to evaluate the impact of The Trivedi Effect®-Consciousness Energy Healing Treatment on the characteristic properties of the phyto-compounds present in the ashwagandha root extract using LC-MS, GC-MS, and NMR spectroscopy. Ashwagandha root extract was divided into two parts, one part of the sample was termed as untreated/control sample, while the other part of the sample received The Trivedi Effect®-Biofield Energy Healing Treatment remotely by a renowned Biofield Energy Healer, Mr. Mahendra Kumar Trivedi and termed as Biofield Energy Treated sample. The LC-ESI-MS data revealed that the chromatographic peak area% of the phytoconstituents present in the treated sample was significantly altered in the range of 31.13% to 20.61% compared to the control sample. Similarly, the average peak area of the treated sample was significantly improved by 20.61% compared to the control sample. A total of 14 withanolides such as viscosa lactone B, dihydrowithanolide D, withanolide A, withanolide B, etc. were proposed with m/z 489, 473, 471, and 455 at the retention times (Rt ) of 13.2, 14.5, 14.8, and 18.2 minutes, respectively. At Rt 13.2 minutes (m/z: 489), the peak area% and mass peak intensity of the phytocompound in the treated ashwagandha were significantly increased by 2.57% and 14.83%, respectively compared with the control sample. Similarly, at Rt 14.5 minutes (m/z: 473), peak area% was increased by 3.76%, but the mass peak intensity was significantly decreased by 26.48% in the treated sample compared with the control sample. At Rt 14.8 minutes (m/z: 471), peak area % was same, but the mass peak intensity was increased by 2.74% in the treated ashwagandha compared with the control sample. Subsequently, at Rt 18.2 minutes (m/z: 455), peak area% was increased 1.01%, whereas the peak intensity was significantly decreased by 10.90% in case of the treated sample compared to the control sample. These findings intensify to assume that The Trivedi Effect®-Consciousness Energy Healing Treatment might be responsible for the overall improvement of relative concentrations and altered relative abundance of the phytoconstituents in the ashwagandha root extract compared to the control sample. This Biofield Energy Treated ashwagandha root extract would be helpful to design better nutraceutical/pharmaceutical formulations against inflammatory diseases, immunological disorders, sexual disorders, arthritis, stress, cancer, diabetes, aging, and other chronic infections.