As the opioid epidemic worsens, forensic labs are called on more often to analyze a variety of samples for drugs of abuse.

The investigation process into the use of drugs or drugs of abuse (DOA) is highly dependent on individual cases. Most often, there is a small shortlist of possible drugs that analysts can screen for in a sample. In such cases, specific and dedicated testing methods are utilized to elucidate the drug involved. The process of analysis also depends on the source of the sample. For example, samples from police institutions usually come with an idea of the type of analyte to be detected: pre-tests in urine have usually been conducted so analysts have a broad idea of the class of drug in question. In these cases, a laboratory only needs to screen for a few analytes. In other cases, such as in the event of intoxication, the drug responsible is less clear and broad screening methods are required. The samples available for analysis also depend on the type of case, and range from blood, urine, hair, vitreous humor and, occasionally, organs.

To analyze drugs and DOA in biological matrices, liquid chromatography mass spectrometry (LCMS) and gas chromatography mass spectrometry (GCMS) are used, but when analyzing body fluids, LCMS has become the choice of method.

Case study: Institute of Forensic Medicine
Volker Auwärter’s laboratory at the Institute of Forensic Medicine, University of Freiburg, benefits from specifically developed toxicological solutions to further research and services in forensic analysis. The laboratory runs drug screening services for organizations across Europe, in Luxembourg, Belgium and Poland. The institute also collaborates with a laboratory in the U.S., where they contribute to method development. As an academic laboratory, the key priorities of the institute’s work lie in research and analytical services, often for requests dealing with legal cases.

The role of the institute is to conduct toxicological analysis on cases where drugs or drugs of abuse are involved. This primarily involves testing blood, urine and hair samples and in post-mortem cases, stomach contents, vitreous humor and occasionally organs such as liver, kidney and muscle tissue. In some instances, analysis of powders, liquids, paper or other materials is also requested. Due to the nature of samples analyzed, semi- or broad-targeted approaches are needed to screen for a vast array of analytes. The source of the samples sent to the laboratory varies, and has changed over time.

“In the beginning, death cases and services for investigative authorities were the main source of work for us as a forensic laboratory. Over time, we have received more samples from the police, taken from offenders under the influence. Then it developed further and we now have more clinical samples, for example from prisons and forensic psychiatric clinics. Now this represents the larger portion of our work,” explains Auwärter.

Currently, the institute receives approximately 11,000 cases to analyze per year, ranging from simple blood alcohol analysis to full post-mortem toxicology. Due to the institute’s focus on research, the majority of funding received by the laboratory is for this purpose.

“The laboratory stands on two legs: one is academic, where we have funding for teaching and research, and the other is the service we provide to private facilities and investigative authorities. The two elements support each other—if we’re good in research, we can offer new, specialized methods to our customers, and if we provide a good service, this generates revenue to channel into innovative research. We’re not a high-throughput laboratory, so we always have to keep on top of the latest developments in order to offer specialized services private laboratories with high-throughput machines are not able to provide,” said Auwärter.

The institute has conducted EU-funded projects since 2011, all of which maintain a core focus on drugs market monitoring and development of methods to detect NPS in biological matrices. The laboratory must apply for new funding every two years, and over time the nature of the project has changed depending on the EU commission’s goals.

“We have to assess what the EU commission wants, and which project would fit in with our purpose. Previously, we have applied for a security research project, as well as the Drug Information and Prevention Program—you always have to look out for those that fit, and often need to adapt to them. In the beginning, the projects were more constructed around questions of prevention, whereas now, the projects are covered more by the criminal investigation area. At the moment, together with the Federal Criminal Police Office, we’re contributing to some profiling work to see which kinds of impurities are found in drugs and learn about their routes of synthesis,” explained Auwärter.

As specialists in NPS detection, the demand for this service from the institute has been high, so the research effort to keep up to speed with rapid developments in identification of new compounds is huge. In 2008, the problem of detection of the NPS “Spice” became a widespread and common occurrence that swept across the industry. As a result, a large portion of the laboratory’s work is now involved with detecting synthetic cannabinoids. Since 2012, screening for NPS has become an issue even in standard cases, and is a constant factor in the laboratory’s work.

“Every death case we complete will undergo the whole spectrum of NPS screening,” adds Auwärter.

The laboratory uses the Toxtyper (Bruker Daltonik) LC-MSn-based library solution for its EU monitoring projects: drug samples are purchased online by the laboratory to be analyzed for the principle active compounds. The instrument quickly identifies these samples and where drugs have already been screened before, it is used to identify specific groups of drugs, for example synthetic cannabinoids, and simultaneously looks for unknown compound spectra. These compounds can then be selected, purified and have their structures analyzed with other instruments.

The robustness, quality of data and ease of use of the instrument have simplified and accelerated the mass spectrometry screening capabilities of the laboratory, shortening the overall turnaround time of a case. By reducing the time spent on routine sample analysis, more time can be spent on complex intoxication or post-mortem cases, which is very valuable. Reduced sample preparation time is one of the key advantages of using such a technique: the time taken to prepare samples for GCMS is up to one hour, whereas this LCMS solution takes approximately 10 minutes.

Moving forward
The designer drug testing arena is rapidly expanding and is unlikely to slow down in the near future. Therefore, laboratories such as the Institute of Forensic Medicine will need to rely more and more on the advancing technologies provided by vendors, which are specifically designed with these research challenges in mind. The commitment of such vendors to provide the innovative instrumentation and industry expertise required for research institutes to carry out increasingly complex work paves the way for future discoveries. The two-way collaboration seen in this case study facilitated the co-development of novel drug screening methods, which enables the institute to further their research and service contributions to the forensic and toxicology industry. 

Example of selected drug detection in whole blood at a concentration of 0.5 ng/mL. Photo: Bruker