Timely detection of myocardial injury is essential for appropriate management of patients in emergency department (ED) evaluated for acute myocardial infarction. A novel electrocardiogram (ECG) metric, the Cardiac Electrical Biomarker (CEB), uses eigenvalue modeling of the 12-lead ECG and quantifies dipolar vs. multipolar forces. The goal of this project was to study association between the CEB and high-sensitivity troponin I (HsTnI). We conducted a retrospective study of patients, evaluated in the ED for acute myocardial infarction [n = 411; 57.6 ± 13.2 years; 186 (45%) men; 266 (64%) African-Americans]. Resting 12-lead ECG and HsTnI were measured at presentation and at 3, 6, and 9 hours after the initial measurement. The CEB was measured by the VectraplexECG System (VectraCor, Totowa, NJ). Patient-specific longitudinal analysis was performed to study association between the CEB with HsTnI changes over time. The CEB indicated myocardial injury in 116 (28.2%) study participants. HsTnI was significantly elevated during ED observation period in patients with myocardial injury, diagnosed by the CEB [median (interquartile range), 10.3 (5.2-31.4) vs. 6.3 (3.5-16.5) ng/L; P = 0.002]. In a mixed-effects linear regression adjusted for age, race, and sex, increasing HsTnI was associated with the CEB elevation [β-coefficient, 0.071 (95% confidence interval, 0.008-0.134); P = 0.027]. In conclusion, in patients in ED evaluated for acute myocardial injury, increasing values of HsTnI were associated with increasing values of the CEB, suggesting that myocardial injury is the mechanism that underlines acute changes in the CEB.