Abstract: The method for extraction and determination of androsterone, 5α-androstane-3α,17β-diol, epiandrosterone and 5α-androstane-3β,17β-diol were developed by SPME with headspace derivatization and GC/MS. It was applied to the simultaneous analysis of 4 anabolic steroids in the spiked human hair with the satisfactory accuracy and precision. The limits of detection(S/N＝3) for 4 anabolic steroids in spiked human hair are in the range of 0.027-0.150μg L^－1.