Background: Cardiovascular diseases are the leading cause of morbidity and mortality globally, with acute myocardial infarction (AMI) being a significant contributor. This study aimed to investigate the roles of cardiac biomarkers, including H-FABP, GPBB, and others, in detecting AMI. Method: Blood samples were collected from 80 individuals, including 50 with AMI and 30 healthy controls, admitted to the Coronary Care Unit (CCU) of an Educational Hospital in Diyala province between May and July 2022. Cardiac markers (hs-troponin-I, myoglobin, CK-MB, GPBB, and H-FABP) were measured using the Sandwich-ELISA technique. Results: There were no significant differences (p>0.05) in age or gender distribution between the study groups. Levels of cardiac markers were significantly higher in AMI patients compared to healthy controls (p<0.05). H-FABP demonstrated the highest sensitivity (100%), followed by GPBB (97%), hs-troponin-I (87%), CK-MB (85%), and myoglobin (78%), with significant differences (p<0.05) in detecting AMI. H-FABP and GPBB also exhibited the highest specificity (98% and 96%, respectively), while myoglobin and CK-MB had lower specificity (82% and 84%, respectively). Furthermore, positive correlations were observed between H-FABP, GPBB, and other markers (hs-troponin-I, myoglobin, CK-MB). Conclusion: H-FABP and GPBB show promise as predictive indicators for early diagnosis (within 1-4 hours of chest pain) of AMI, offering potential utility in clinical practice.