Determination of postmortem interval from old skeletal remains by image analysis of luminol test results

The luminol test is routinely used in forensic serology to locate blood traces and identify blood stains not visible to the naked eye; its sensitivity is reported as ranging from 1:100,000 to 1:5,000,000. To evaluate the possibility of correlating the postmortem interval with blood remnants in bone tissue, the luminol test was performed on 80 femurs with a known time of death, grouped in five classes. Powdered bone (30 mg) was recovered from compact tissue of the mid-shaft of each femur and was treated with 0.1 mL of Luminol solution (Sirchie Finger Print Laboratories, Inc.). The reactions were observed in a dark room and filmed by a TV camera equipped with a recording tape. An intense chemiluminescence was observed after a few seconds in all 20 femurs with a PMI ranging from 1 month to 3 years. On the 20 femurs with a PMI ranging from 10-15 years, a clear chemiluminescence was visible with the naked eye in 80% of the sample. Among the 20 femurs with a PMI ranging from 25 to 35 years, a weaker chemiluminescence appeared in 7 femurs (33% of the sample). In the 10 femurs with a PMI ranging from 50 to 60 years, a faint reaction was observed only in a single femur. In none of the ten femurs with a PMI over 80 years was chemiluminescence observed. The image of each reaction was computerized and analyzed for gray scale. The results of image analysis show a possible quantitative relationship between the PMI and luminol chemiluminescence in powdered bone.