Identification and Prevalence Determination of Novel Recreational Drugs and Discovery of Their Metabolites in Blood, Urine and Oral Fluid

Designer drug products which contain a variety of unregulated psychoactive constituents (phenethylamines, cathinones, and synthetic cannabinoids) have become main stream on the illicit drug market and are highly prevalent at electronic dance music festivals (EDM). We propose the collection and analysis of paired blood, urine, and oral fluid (OF) samples from volunteer participants attending these festivals, many of whom are likely to have ingested some of the newest compounds on the market. This proposal presents an opportunity to obtain information regarding what new drugs are on the market, their prevalence, identification of their metabolites, and correlations between their presence in blood, urine and OF specimens, allowing insight into patterns of use. Limited data exists in the literature comparing blood and OF test results. OF is an important, emerging matrix that can be collected rapidly, non-invasively, and near the time of a persons drug use or apparent intoxication. The ability to compare blood and OF results in addition to correlating these values with self-reported drug use, will allow elucidation of the relative value of these two matrices in assessing recent drug use. Additionally, inadequate existent research fails to establish the identity of metabolites of many novel designer drugs. Rapid changes in the drug market of synthetic compounds cause epidemiological studies to be completed long after drugs are introduced to users, and the scope of testing frequently fails to detect, identify or report the most recently available drugs. This project will collect paired blood and urine samples, which will allow for the possibility to connect parent compounds in blood with putative metabolites in blood and urine. Using authentic samples allows these determinations to be made in vivo, without dosing subjects and avoiding problems encountered in in vitro studies where the metabolic profile often differs from those in the intact organism. Identification of these metabolites would give insight to manufacturers of standard reference materials to improve availability of commercial standards, allowing more laboratories to identify their use in at risk criminal justice and public health settings. Samples will be collected over the days of March 14-23, 2014 at the Ultra Music Festival, an EDM festival in Miami, Florida. Following verbal recruitment of participants, volunteers will enter the study site, which is designated a safe zone, free of drug enforcement activity. Participants will have the study explained to them in full, asked to sign an informed consent form, and given a unique identifier number, which will be used as the only form of identification of all samples collected, allowing samples to be linked to one another, but not to the donor. Blood will be collected by a trained phlebotomist, and urine collection will occur in a private lavatory facility on-site. OF will be collected and tested on-site for the presence of priority drugs of abuse, and a secondary specimen will be collected for laboratory confirmation. Blood, urine and collected OF will be evaluated by screening for a broad range of therapeutic, abused and designer drugs currently comprising 327 compounds, using Liquid Chromatography-Time of Flight Mass Spectrometry. The scope is likely to be expanded due to changes in the illicit drug market between now and the time of the study. Confirmation for all detected compounds will occur using either the Liquid Chromatography Tandem Mass Spectrometry, or Gas Chromatography Mass Spectrometry, as appropriate. Novel artifacts and metabolites will be identified by comparison between samples containing parent drug (blood and oral fluid) and urine, using available extensive in-house, commercial and public mass spectral databases. Additional method development and validation may be required upon the detection of previously unidentified compounds and their metabolites. ca/ncf