In the present study liquid-liquid extraction (LLE) of amphetamine from biological samples was systematically optimised. The recovery of amphetamine extraction was studied using a variety of 5 solvents (chloroform, 1-chlorobutan, dichloromethane, diethylether, ethyl acetate) as well as a system of solvents (chloroform: ethyl acetate: ethanol, 3:1:1 v/v). Furthermore the influence of the volume of the extracting solvent and the pH on the recovery of amphetamine was examined. The optimum conditions of LLE procedure were applied for the extraction of amphetamine from biological samples such as urine, blood and also from less commonly used biological specimens such as liver, bile, brain, vitreous humour, pericardial fluid, cerebrospinal fluid, bone marrow and bone. The determination of amphetamine was accom- plished by gas chromatography equipped with a flame ionization detector (FID), after derivatization with heptafluorobu- tyric anhydride (HFBA).