An extended and comprehensive review is presented herein, focusing on sample preparation (pretreatment and extraction) and different analytical methods applied for the quantification of tricyclic antidepressants. These procedures are relevant tools in clinical and forensic toxicology. It is revealed that SPE, for sample preparation, and HPLC, using reversed-phase alkyl (C18) or cyanopropyl-bonded silica columns for the analytes separation, are effective and versatile methods for assay of tricyclic antidepressants. These methods enable achievable detection limits using UV/diode array detection, readily available in most laboratories, down to 1-8 ng ml(-1), and using electron capture detection better than 1 ng ml(-1), which is lower than that for nitrogen-phosphorus detector. MS interfaced with electrospray ionization offered similar sensitivity, whilst sonic spray ionization provided detection down to 0.03 ng ml(-1). A brief discussion on chemical structures, metabolism and mechanism of action of this group of drugs is also presented.