Purpose. The Federal Aviation Administration's Office of Aerospace Medicine (OAM) is responsible for the certification of pilots with diabetic conditions. Therefore, it is essential for OAM to monitor pilots involved in fatal accidents for abnormal glucose levels, which might have caused performance impairment/ incapacitation. The present study evaluated the use of postmortem vitreous humor and urine glucose levels in transportation accident fatalities as indicators of potentially incapacitating medical conditions or performance impairment.Methods. Vitreous humor and/or urine from 192 accident fatalities were analyzed for glucose using a hexokinase method. Cases with values below the lower limit of detection (10 mg/dL) and above 3 standard deviations (SD) from the mean were not included in the final statistics. All cases more than 5 SD above the mean were deemed abnormal and a full case history was evaluated based on the available medical history.Results. The mean glucose concentration was 30 mg/dL (SD: 21 mg/dL) from 98 postmortem vitreous humor specimens, while it was 27 mg/dL (SD: 16 mg/dL) from 127 postmortem urine samples. Nine of the 192 cases were identified as having abnormal glucose levels. Abnormal glucose levels were found in 5 of the 8 cases with a known diabetic condition. Glycosuria or low renal threshold was reported in 2 fatal pilots; 1 of these pilots had an abnormal glucose level.Conclusions. Hyperglycemia can be established from the vitreous humor and urine glucose levels, but hypoglycemia cannot, because of the rapid postmortem drop in vitreous humor glucose levels. All of the pilot abnormal glucose cases detected were previously identified during the medical certification process or had a medical reason for the abnormal level. The elevated vitreous humor and urine glucose levels have proven useful in identifying individuals with a pre-existing diabetic condition that might have been a factor in the accident.