Extraction and Analysis of a Comprehensive Quantitative Drug Panel in Hair Samples by UPLC-MS/MS for Forensic Toxicology

The detection and quantification of drugs in hair presents many challenges, especially when analyzing multiple classes of drugs simultaneously. The objective of this work was to develop, optimize and validate a method for the extraction and quantification of a comprehensive panel of drugs in hair to satisfy the confirmation cut-off values recommended by the society of Hair Testing (SoHT).
Sample preparation conditions were adjusted for optimal extraction of all compounds, including hair pretreatment, the initial incubation conditions and solid phase extraction. The finalized method included an initial incubation to extract analytes from hair samples, followed by mixed-mode SPE clean-up and UPLC-MS/MS analysis using the Waters TQ Absolute. The drug panel included, opiates, amine stimulants, benzodiazepines, synthetic opiates and other common drugs of abuse.
Efficient and clean sample extraction enabled a robust method that was validated for linearity, sensitivity, accuracy and precision for 58 analytes. We were able to achieve functional sensitivities down to 0.01 ng/mg for most analytes and as low as 0.001 ng/mg for fentanyl allowing all analytes to meet their SoHT recommended cut-offs. All analytes in external positive control samples were positively identified and >80% had concentrations that fell within the manufacturer’s acceptance range.
We have developed a robust method for quantitative analysis of a comprehensive panel of drugs in hair that meets SoHT’s cutoff criteria. This enabled the quantitative validation of a method for 58 compounds that can be applied to other related compounds as well.
Key learning objectives:
- Understand the challenges with detecting and quantifying multiple classes of drugs in hair samples
- Learn how to develop a comprehensive multi-panel drug method in hair samples that meets SoHT's cutoff criteria
- Learn detailed sample preparation techniques, including hair pretreatment and solid phase extraction
For Forensic Toxicology Use only.
Presenter: Jonathan P Danaceau
Dr. Jonathan Danaceau, Ph.D. is a Principal Scientist with Waters. He received his B.S. in Biology from Allegheny College and a Ph.D. in Neuroscience from the University of Utah (Salt Lake City, UT). Jon came to Waters in 2011 after spending several years in the forensic toxicology and sports doping fields. Since joining Waters, he has focused on sample preparation and LC/MS/MS solutions for forensic, clinical research, and other bioanalytical applications. He is an expert in sample preparation and bioanalytical method development and has published several papers in these fields. Jon is currently with the Waters Clinical Business Unit.
