Hair has been focused on for its usability as an alternative biological specimen to blood and urine for determining drugs of abuse in fields such as forensic and toxicological sciences because hair can be used to elucidate the long intake history of abused drugs compared with blood and urine. Hair analysis consists of several pretreatment steps, such as washing out contaminates from hair, extraction of target compounds from hair, and cleanup for instrumental analysis. Each step includes characteristic and independent features for the class of drugs, e.g., stimulants, narcotics, cannabis, and other medicaments. In this review, recently developed methods to determine drugs of abuse are summarized, and the pretreatment steps as well as the sensitivity and applicability are critically discussed.