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Power of High Res Mass Spec to Detect Fentanyl Analogues
Thursday, July 27, 2017 2:45:00 PM UTC - 4:00:00 PM UTC
Duration: 1 hour(s) 15 minute(s)
This webinar summarizes the pharmacology of Non-Pharmaceutical Fentanyls (NPFs), trends of abuse, and explore novel techniques for the analysis of NPFs in forensic toxicology casework using Q-TOF mass spectrometry applications.
Webinar Time: 11:00am EDT/8:00am PDT/3:00pm GMT

Learning Objectives:

1)  Describe the pharmacology of Non-Pharmaceutical Fentanyls (NPFs) and their current trends of abuse in forensic toxicology.

2)  Create an analytical scheme using various data acquisition modes to identify unknowns such as NPFs in forensic toxicology casework using QTOF instrumentation. 

This webinar will summarize the pharmacology of NPFs, trends of abuse, and explore novel techniques for the analysis of NPFs in forensic toxicology casework using Quadrupole Time-of-Flight (Q-TOF) mass spectrometry applications.

The number of opioid overdoses in the United States has dramatically increased over the past few years.  The abuse of heroin and synthetic opioids such as fentanyl and its analogues are major contributors to this epidemic.  Non-pharmaceutical fentanyls (NPFs) such as carfentanil, furanyl fentanyl, acetyl fentanyl, fluoroisobutyryl fentanyl, and other novel opioids such as U-47700 and 4-ANPP are being routinely encountered in forensic toxicology and drug chemistry casework.  Furthermore, we will highlight prevalence statistics from Alabama, Palm Beach County, and other laboratories.

Immunoassay-based techniques with antibodies targeted to fentanyl typically have some level of cross-reactivity to NPFs due to chemical structure similarity.  However, false negatives may persist if confirmation methods only target fentanyl and metabolites.  More sophisticated methods and instrumentation may be required to adequately detect NPFs.  Laboratories should strive to incorporate new forensic toxicology and drug chemistry applications to stay current with emerging novel psychoactive substances. 

Forensic toxicology applications focused on a methodology for the detection and confirmation of NPFs in whole blood will be discussed.  An Agilent 6545 Q-TOF Mass Spectrometer (MS) was used to conduct high resolution mass spectrometry screening and fragment confirmation. An in-house library was constructed for the purposes of screening by accurate mass extraction and confirmation by fragmentation spectra.  The library was created by the addition of empirical formulas for currently available NPFs and other novel opioids for accurate mass calculation.  Available standards were then purchased and their retention times and fragmentation spectra were added to the library.

Screening and confirmation by the Q-TOF mass spectrometer was accomplished by conducting two experiments with extracts from a liquid-liquid extraction.  The first injection was for MS only full scan data collection in TOF mode and the data processed by an accurate mass extraction and scoring.  General scoring criteria for MS mode were based on a scoring algorithm of extracted ion chromatograms that included mass accuracy, retention time, isotopic spacing, and isotopic abundance.  The second injection was for MS/MS mode with varying collision energies for fragmentation of the accurate mass precursor ions.  Agilent MassHunter Acquisition software allows for operation of the Q-TOF in three different MS/MS modes: All Ions, Auto MS/MS, and Targeted MS/MS.  The confirmation capability of the TOF screen results for the different modes was compared.  In addition, general method development strategies will be presented to include chromatography, MS and MS/MS mode operational settings, and data processing algorithms that were used for this methodology.

For Forensic Use


Curt Harper & Jason Hudson
Curt Harper - Appointed Toxicology Discipline Chief for the Alabama Department of Forensic Sciences (ADFS) in 2012. 

Jason Hudson - Forensic Toxicology Section Chief at the Alabama Department of Forensic Sciences, Birmingham Laboratory.