Metabolites identified for the novel psychoactive substances (NPS) alpha-pyrrolidinophenone (alpha-PVP), methylone, and dimethylone were identified using an in vitro process and subsequently compared to metabolites produced in vivo from human drug-user samples to determine the extent to which each metabolite could be detected in authentic biological specimens of recreational users and which metabolites would serve as the most valuable biological markers of use.
The widespread use of NPS at electronic dance music (EDM) festivals has been well documented, and several adverse events and fatalities from the ingestion of these recreational drugs have been reported in the United States. The diversity and rapid turnover in the prevalence of any particular NPS at any given time has created several challenges for public health officials, law enforcement, and forensic science communities. The current study demonstrated the value of using oral fluid as a specimen for drug detection compared to blood. Through the use of in vitro metabolism studies, metabolic pathways for alpha-PVP and dimethylone were proposed. These metabolites were subsequently identified in authentic blood, urine, and oral fluid specimens. The most prevalent metabolite for alpha-PVP was the 5-OH-PVP metabolite and for dimethylone were methylated dimethylone in blood and oral fluid and the hydroxylated dimethylone in urine. In a few cases, the parent drug was not confirmed; however, the presence of unique metabolites could be used in many cases to indicate which parent drug the subject had ingested. During 2013 and 2015, biological samples were collected from 396 individuals (126 blood samples; 227 urine samples; 122 oral fluid samples screened with the Alere DDS2; and 384 oral fluid samples collected with the Immunalysis Quantisal oral fluid collector). 27 figures, 26 tables, and appended extensive tabular data