Like traditional opioids, these novel synthetic opioids are abused due to their analgesic and euphoric effects; however, their adverse side effects include respiratory distress, nausea, and decreased consciousness. Comprehensive analytical methods for detecting and quantifying these opioids are limited. The comprehensive analytical method used in the current project for the detection and quantification of these drugs was a solid-phase extraction followed by liquid chromatography- tandem mass spectrometry (LC-MS-MS). This method was validated according to the Scientific Working Group for Forensic Toxicology (SWGTOX) guidelines. Four lots of pooled oral fluid (n=5 contributors) were used as different sources for the required parameters. Linearity was determined by using six non-zero calibrators over 5 days. Limit of quantitation was evaluated as the lowest non-zero calibrator (10 ng/ml) in duplicate in three different lots over 3 days. Bias and precision were assessed in triplicate at low, medium, and high QC concentrations over 5 days. To reduce matrix effects and improve internal standard response over the linear range, reconstitution volume was optimized to 1 ml. To the knowledge of the researchers, this is the first publication with a validated method for extraction and quantification of W-series drugs. Although there are limited studies that involve detection of NSO, particularly in oral fluid, the LOQs validated in this study are anticipated to be sufficient for authentic samples. This method is currently being used to determine drug trends in several groups. Analysis of oral fluid specimens from sensitive populations that may be seeking alternative or substitute opioid use will enable examination of the prevalence of novel synthetic drugs in the market. 5 tables and 42 references
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