The present study seeks to find a way to quickly and correctly differentiate myocardial infarction from unstable angina by measuring the creatine kinase-MB/creatine phosphokinase ratio and comparing in non-ST elevation myocardial infarction patients with unstable angina at different time intervals, to improve the health quality of patients with coronary artery disease. The present study is a retrospective epidemiological analysis of 260 patients with non-ST elevation myocardial infarction and 260 patients with unstable angina, including age, sex, creatine kinase-MB, and creatine phosphokinase biomarkers at two-time intervals, including referral (4-8 h from the onset of pain) as the first interval, and 8 h after the first sampling was extracted as the second interval. Moreover, the delta of the creatine kinase-MB/creatine phosphokinase ratio during two interval times was measured. In non-ST elevation myocardial infarction patients in the first and second intervals, creatine kinase-MB/creatine phosphokinase ratio was 32.7 and 33.8% higher than the normal laboratory cutoff (positive), respectively, and in the group of unstable angina patients, this index was positive in 31.9 and 30.4% of patients, respectively. There was no significant difference between the mean creatine kinase-MB to creatine phosphokinase index between the patients with non-ST elevation myocardial infarction and unstable angina (p = 0.507). In the first interval, the sensitivity and specificity of this index in differentiating non-ST elevation myocardial infarction from unstable angina were 51.5 and 57.3% (area under the curve = 0.518), respectively. While in the second interval, the sensitivity and specificity of this index were 17.7 and 87.8% (area under the curve = 0.519), respectively. The creatine kinase-MB/creatine phosphokinase delta in the non-ST elevation myocardial infarction group was significantly higher than in patients with unstable angina during different time intervals (p = 0.01). According to our results, creatine kinase-MB/creatine phosphokinase index cannot help differentiate the two groups of non-ST elevation myocardial infarction and unstable angina. However, the findings show that higher levels of creatine kinase-MB enzyme and creatine kinase-MB/creatine phosphokinase delta in the early hours, 4-16 h after the onset of pain in non-ST elevation myocardial infarction patients, can be used to differentiate between non-ST elevation myocardial infarction and unstable angina.