Methodology using MAE/SPME/GC–MS is being pursued for the analysis of organic pollutants in sebum. The microwave-assisted extraction (MAE) of standards of semi volatile organic pollutants from sebum was optimized. All compounds were extracted from sebum with recoveries analyzed by GC/MS ranging from 94% to 100% under the optimum MAE conditions: 10mL acetone–hexane (2:1), 60°C, and 10min microwave heating. To improve the detection limits a SPME procedure was optimized. Linearity ranged from 0.70ppb to 25ppb. R.S.D. were in the range of 1–23% for the SPME step. Preliminary real samples were analyzed and a range of compounds was detected. The optimized MAE/SPME/GC–MS methodology promises to be useful for different applications.