by Joshua DeBord, Senior Scientist, CFSRE, and Chris Elicone, Director of Product Management, Bruker
The global novel psychoactive substances (NPS) crisis is a significant, and growing, international public health threat. Sharing similar effects with established illicit drugs like fentanyl and alprazolam, NPS are synthetic substances that are designed to evade legal control through manufacturers regularly altering their chemical structures.
There is limited data on the safety, toxicity and long-term health consequences of many NPS, and their frequent modification and growing number makes it increasingly difficult for forensic and clinical laboratories to keep pace. By July 2024, 1,245 individual NPS had been reported to the United Nations Office on Drugs and Crime (UNODC) from across the globe, and there is growing concern of further proliferation of nitazenes, a group of synthetic opioids responsible for clusters of fatal overdose events.
The U.S. Center for Forensic Science Research and Education (CFSRE) has established a proactive early warning system under the NPS Discovery program, whose extensive research has provided forensic professionals, from local crime laboratories to those in federal agencies, with valuable trend reports, drug alerts and forensic monographs to help fight the problem through knowledge. The program analyzes the chemical composition of samples from across the country to support trend investigation at a regional and national level.
However, the performance of the NPS Discovery program is only as good as the speed and reliability of analysis in the forensic lab, where detection techniques can lag behind new variants.
Researchers at the CFSRE have been working to develop faster and more flexible methods to detect and identify NPS in biological and drug samples. A new chromatography-free Direct Analysis in Real Time (DART-MS/MS) workflow based on an advanced mass spectrometry instrument that can run both DART and liquid chromatography-mass spectrometry (LC-MS/MS) on the same system is set to provide a new solution in the field of NPS detection and quantification.
The analytical challenge of NPS
Because NPS tend to have a short lifespan in the drug market, acquiring accurate and timely information can be difficult. The issue is compounded by the fact that, once the details are available, that substance may no longer be the greatest concern. Events associated with NPS are also relatively localized, making it even more challenging to respond rapidly and effectively to each individual sample.
Traditional forensic screening methods, such as colorimetric testing and immunoassays, require lengthy development and validation, can be cost-prohibitive, inflexible and frequently fall short in performance—lacking specificity, producing false negatives for NPS and offering inadequate sensitivity for highly potent compounds at low concentrations. As a result, many forensic chemistry units are investigating the feasibility of alternative technologies.
The forensic community currently relies upon gas chromatography-mass spectrometry (GC-MS) for drug sample analysis, which can lack the sensitivity required to screen for NPS. While a typical preparation of methamphetamine or heroin produces a sample suitable for GC-MS, the more challenging NPS may not appear on an initial screen. For example, synthetic opioids, such as carfentanil and nitazene analogs, as well as benzodiazepines, occur at levels within mixtures that can be challenging to detect by GC-MS. Where a timely response is of the essence, labs are under increased pressure to find a rapid and robust solution.
The DART advantage
Enter chromatography-free technology. Its versatility, ease of use and the ability to run both DART and LC-MS/MS on a single mass spectrometry system enables analysis in greater detail than with other systems. By first screening suspected drug samples with GC-MS and then moving to a DART workflow to help confirm challenging NPS, same-day results are possible.
DART mass spectrometry is a chromatography-free, high-productivity ambient ionization technique that analyzes samples quickly and with minimal sample preparation. Samples are ionized and analyzed immediately by a mass spectrometer. The approach has enabled the NPS Discovery program to increase sample throughput while performing analysis in real time has made method development more efficient.
Supporting nitazene detection
Nitazenes are a group of synthetic opioids which were initially developed as an alternative to morphine but were never medically approved due to their extreme potency. They are notoriously difficult to detect as they can be mixed with other substances at minute amounts (often lower than 1 percent mass). In 2019, the first nitazene was reported to UNODC and a further 26 substances have been identified in the years since.
The NPS Discovery laboratory is working to develop a new method for the fast and accurate identification of nitazene substances. The lab has historically used GC-MS as a screen, but it was challenging to detect low-abundance mixture components like nitazene analogs. The sensitivity of DART allows the lab to explore the similar fragmentation of nitazene analogs to assist with identifying truly unknown NPS. In as little as eight seconds, DART can identify the presence of nitazene analogs in a sample, even in mixtures where they would have gone undetected by GC-MS analysis.
Improving frontline screening
The CFSRE is considering replacing its current GC-MS front-line screening technology with the DART-MS chromatography-free workflow to help detect challenging NPS such as benzodiazepines and synthetic opioids at an earlier stage, with the goal of facilitating faster response times. Workflow automation using the chromatography-free approach, and integrated robotic sample preparation are also seen as further valuable system developments. Because of the large quantity of data generated, the team is working toward automating data review to help achieve the highest possible throughput, with the ultimate goal of achieving a flexible, rapid analytical response to the NPS crisis.
About the authors
Dr. DeBord is a Senior Scientist at the CSFRE where he oversees the chemistry arm of the NPS Discovery research program. Following a PhD in Analytical Chemistry investigating heroin provenance through chemometrics, Dr. DeBord transitioned into forensic science through a toxicology and drug chemistry role before joining the CSFRE in 2022. Following more than 10 years in the forensics field, he leverages forensic and data science to support public safety investigations and public health research.
Christopher Elicone is Director of Product Management at Bruker, with more than 25 years of experience advancing mass spectrometry technologies and analytical workflows. His work spans diverse applications, including forensic toxicology, clinical research, food safety, environmental analysis and pharmaceutical testing. Chris worked on innovations across a range of platforms—from portable ion trap systems to GC and LC triple quadrupoles and high-resolution mass spectrometry. He holds a Chemistry degree from Emory University in Atlanta, GA.