Cardiac biomarkers are important targets for any test sample to discover ideal cardiac health supplement. The study was aimed to evaluate the cardioprotective activity of Biofield Energy Treated test sample (DMEM) using rat cardiomyocytes (H9c2) cells. The test item (DMEM medium) was divided into three parts, first part received one-time Biofield Treatment by Alice Branton, a renowned Biofield Energy Healer, and defined as the one-time Biofield Energy Treated (BT-I) DMEM, while second part received two-times Biofield Energy Treatment and denoted as the BT-II DMEM. The third part did not give treatment and denoted as the untreated DMEM group. The cell viability of the test sample by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay showed an increase level of viable cells by 86.58% and 91.37% in the BT-I and BT-II groups, respectively. Besides, BT-I and BT-II groups showed 10.56% and 23.53% cytoprotective action compared to the tert-Butyl hydroperoxide (t-BHP) induced group. Further, lactate dehydrogenase (LDH) level was significantly (p≤0.001) reduced by 54.43% and 53.52% in the BT-I and BT-II groups, respectively compared to t-BHP induced group. Moreover, the level of creatine kinase-myocardial band (CK-MB) was reduced by 63.48% and 60.68% in the BT-I and BT-II groups, respectively compared to the t-BHP induced group. Additionally, reactive oxygen species (ROS) level was significantly inhibited by 55.6% and 112.8% in the BT-I and BT-II groups, respectively compared to the t-BHP induced group. Besides, apoptotic activity was significantly inhibited by 16% and 10% in the BT-I and BT-II groups, respectively compared to the t-BHP induced group. Overall, the experimental data demonstrated that The Trivedi Effect® has significant capacity to improve the overall cardiac parameters in H9c2 cells, which can be used against various cardiac disorders such as congestive heart failure (CHF), cardiomyopathy, heart valve disorders, arrhythmias, myocardial infarction, etc.