This paper presents a rapid and sensitive method for the determination and chiral separation of amphetamines and related designer drugs in hair samples. The substances are extracted from hair matrix by a 30 min treatment with a saturated carbonate buffer at pH 10 under ultrasonication. A commercial chiral derivatizing agent, trifluoroacetyl-prolyl chloride, is then added to the solution that is directly extracted with hexane and subsequently analyzed by GC/MS in SIM mode. R and S isomers of amphetamine, methamphetamine, MDA, MDMA and MDEA can be separated and detected with a limit of detection of 0.1 ng/mg for amphetamine, methamphetamine and MDA, and of 0.2 ng/mg for MDMA and MDEA. The method was then applied to 12 samples from suspected amphetamines abusers, showing the presence of both isomers of amphetamine and MDMA in one sample (27 and 1.5 ng/mg, respectively) and of MDMA in further eight samples, in concentrations ranging from traces to 2.7 ng/mg. No differences were observed in the disposition of different isomers in hair.