Reported incidences of drug-facilitated sexual assault (DFSA) are on the increase worldwide. These cases represent a particular challenge for the forensic toxicologist due to the difficulty in obtaining adequate evidence of drug administration. Primarily, this is due to the nature and diversity of the drugs involved, their pharmacology and sampling timescales. Evaluating whether a drug has been administered to a victim for the purpose of sexual assault can often be difficult, if not impossible. This review draws attention to this burgeoning crime and focuses on the unique challenges DFSA cases present in terms of evidential analysis. Current analytical methodologies for investigating DFSA are highlighted and discussed along with developments in improving analytical procedures. In particular, enlarging detection windows by adopting emerging LC-MS techniques is also discussed. This review also highlights the use of cutting-edge technologies such as ultra-HPLC and the use of alternative matrices for addressing the problem of improved retrospective drug detection.