Dried blood spot (DBS) is a sampling technique in which a small volume of blood is collected on a specific type of filter paper. Since the 1960s, DBS has been primarily used for the newborn screening of metabolic diseases. With the development of highly sensitive analytical instruments, the dried blood spot technique allows for accurate quantification of ethanol, stimulants, drugs of abuse and heavy metal elements in biological samples. Due to its obvious advantages, the application of DBS in forensic toxicology has witnessed significant growth in the past decade, which is highlighted and presented in this article. Some challenges and suggestions of dried blood spot applications were summarized for the further research as well.