Myoglobin has been described as an early marker of myocardial injury. It increases within 1 to 3 hours of myocardial injury, and falls back to normal early after the event. Few data suggest that myoglobin can be used to triage patients with chest pain. To assess the diagnostic utility of second myoglobin levels drawn within 2 to 3 hours after presentation to the emergency department for detecting myocardial infarction, we prospectively collected myoglobin levels in 368 patients aged > or = 30 years who were admitted with chest pain. Myoglobin levels were measured at admission and 2 to 3 hours later. Sensitivity and specificity for detecting acute myocardial infarction for a twofold increase in myoglobin level from baseline were 39% and 98%, respectively. The early diagnostic performance of myoglobin at admission, the second level drawn 2 to 3 hours later, and creatine kinase-MB mass drawn at admission were similar (receiver-operating characteristic curves 0.80, 0.86, and 0.85). The diagnostic performance of each of these markers was significantly superior to the absolute change from baseline to second myoglobin (receiver-operating characteristic curve 0.77). In patients who presented within 4 hours of symptom onset, myoglobin drawn 2 to 3 hours later had the highest yield for detecting myocardial infarction. These results suggest that serial myoglobin measurement aiming to detect changes over time is not superior to single marker determinations. Myoglobin measured in 2 to 3 hours from admission may be helpful in triaging patients who present within 4 hours from onset of symptoms.