FORENSIC TOXICOLOGY - Bio Medical Forensics

K11 Simultaneous Screening and
Confirmation of Drugs in Biological
Fluids Utilizing LC/MS/MS
Tania A. Sasaki, PhD*, Applied Biosystems, Inc., 850 Lincoln Centre
Drive, MS 430, Foster City, CA 94404
After attending this presentation, attendees will understand a simple
method used to detect and confirm the presence of drugs of abuse and
their metabolites in various biological matrices. This method has very
simple sample preparation and can detect and identify drugs across
several different compound classes.
This presentation will impact the forensic community and/or
humanity by demonstrating the ability of toxicologists to screen samples
in a simpler and quicker manner. They also will have the capability to
screen across several drug classes in a single experiment.
Rapid detection, identification, and quantification of drugs in
biological matrices are important aspects of forensic toxicology.
Typically, GCMS, HPLC, immunoassays, TLC, and various other
methods are used to screen for drugs and GC/MS is used confirmation
of drugs in Forensic analysis. The use of LC/MS/MS for screening,
confirmation, and quantitation of drugs in toxicological assays is
becoming increasingly common due to the simplicity, selectivity, and
sensitivity of the technique.
A simple LC/MS/MS method was developed to analyze biological
fluids (urine, blood, and oral fluids) for hundreds of common drugs of
abuse and/or their metabolites, including opioids (including Fentanyl),
sympathomemetic amines, antidepressants, benzodiazepines, cocaine,
and THC. A hybrid triple quadrupole/linear ion trap mass spectrometer
was used for detection, which allowed confirmation using full scan
MS/MS spectra and quantitation using multiple reaction monitoring
(MRM). Detection limits for all analytes can be as low as pg/mL range.
Sample preparation was eliminated or greatly simplified versus
analogous experiments using other chromatographic techniques and
experimental run times were on the order of 10 - 15 minutes.
Toxicology, Drug Screening, LC/MS/MS
K12 A Novel Method to Extend the Detection
Window of Drug Administration in Victims
of Malignant Assault With Hybrid
LC/MS/MS Technology Combining Triple
Quadrupole and Ion Trap Technology
Andre Schreiber, PhD*, Applied Biosystems/MDS Sciex, 71 Four Valley
Drive, Concord, Ontario L4K4V8, Canada
After attending this presentation, attendees will understand the
advantages of using hybrid triple quadrupole linear ion trap mass
spectrometry to identify phase I and phase II metabolites of drugs.
This presentation will impact the forensic community and/or
humanity by helping to determine whether a drug has been administered,
even after the parent drug has been completely eliminated from the
victim’s body.
A research method has been developed to detect drug intake long
after a dose has been administered. This is achieved by detecting
specific Phase I and Phase II metabolites that are continually excreted
post dose, far longer than the parent drug. Drugs and metabolites are
detected in positive mode utilizing specific Multiple Reaction
Monitoring (MRM) experiments. Information-dependent criteria for
acquisition of an enhanced product ion (EPI) scan result in precursor ion
fragmentation to characteristic product ions. Fragmentation occurs at
varying collision energies and enables spectral comparison to drug
libraries. In addition, Phase II metabolites, namely glucuronides are
detected using true Neutral Loss (NL) scanning and identified by EPI
acquisition and spectral matching. The loss of dehydroglucuronic acid
with a m/z ratio of 176 is characteristic of all glucuronide metabolites.
Chromatographic separation is based on a 2.1 mm ID, 5 micron Gemini
column with an acetonitrile, formic acid, and ammonium formate mobile
phase gradient ramp optimized for separation of various drugs and
metabolites. The method is used to detect drugs in forensic and clinical
research samples and was developed to provide greater scope,
sensitivity, and selectivity compared to conventional methods of drug
detection. The method will help to determine whether or not a drug has
been taken/administered even after the parent drug has been completely
eliminated from the body of the victim.
LC/MS/MS, Metabolites, Toxicology
K13 Application of Ion Mobility Spectrometry
to the Analysis of Gamma-Hydroxybutyrate
and Gamma-Hydroxyvalerate in
Toxicological Matrices
Jennifer W. Mercer, BS*, West Virginia University, 217 Clark Hall,
PO Box 6045, Morgantown, WV 26506; Diaa M. Shakleya, PhD,
National Institute on Drug Abuse, Chemistry and Drug Metabolism
Section, 5500 Nathan Shock Drive, Baltimore, MD 21224; and
Suzanne C. Bell, PhD, West Virginia University, 217 Clark Hall,
PO Box 6045, Morgantown, WV 26506
After attending this presentation, attendees will learn about a rapid,
portable, screening technique for the simultaneous analysis of GHB,
GHV, and analogs in urine. The physical extraction of the hydrophilic
analytes from urine will be discussed, as will the benefits of ion mobility
spectrometry in forensic analyses.
This presentation will impact the forensic community and/or
humanity by introducing a physical extraction with ion mobility
spectrometry as a rapid, portable screening technique suitable for the
detection of GHB, GHV, and analogs in urine.
The predator drug, gamma-hydroxybutyrate (GHB), the lactone
precursor (gamma-butyrolactone, GBL), and the diol precursor (1,4butanediol,
BD) continue to present significant analytical challenges to
forensic toxicologists and chemists. The five-carbon analog (gammahydroxyvalerate,
GHV) and the corresponding lactone (gammavalerolactone,
GVL) are emerging as substitutes for GHB, adding
further complications.
A rapid and reliable screening test for detection of GHB and GHV
would be useful for toxicologists as well as forensic chemists working
with solid dose samples. This lab has previously reported a microcrystal
test effective for aqueous solutions, but felt the development of a rapid,
simple instrumental test effective for screening urine required
development. In addition, GHB and GHV are extremely hydroscopic
and hydrophilic, negating the possibility of rapid and simple extractions
that might be necessary for quick screening. Thus, any successful
GHB/GHV screening methodology must either be matrix independent or
insensitive or capable of rapid and semi-quantitative extraction from the
matrix. The latter issue has been the limiting factor to date.
Ion mobility spectrometry (IMS) was investigated as a method of
screening urine for the presence of these drugs and their degradation
products. In the present study, a high-performance split/splitless injector
and autosampler were utilized to effect a physical separation of GHB
and GHV from aqueous matrices (including urine) based on differences
in relative volatility. This was achieved by a timed period of solvent
evaporation followed by rapid temperature increase and thermal
desorption of the residuals. The injection method in effect replaces
problematic solvent extraction methods with a physical extraction, an
efficient method in the present case considering the hydrophilic nature
of GHB. Sample was introduced directly into a detection system
129 * Presenting Author