FORENSIC TOXICOLOGY - Bio Medical Forensics

Eurachem method. Validation results established that routine
quantitative amitriptyline and qualitative morphine determination can be
achieved in liver and larvae matrices. Also at this study; a formula was
suggested for a back-calculation from the results that were obtained from
the decomposed liver matrix and larvae collected at the end of the
117(±0.5) hours period, to the 0 th hour liver concentration.
Entomotoxicology, Tricyclic Antidepressants, Opiates
K17 An Application of Speciated Isotope
Dilution Mass Spectrometry (SIDMS) for
Simultaneous Drug Quantitation of
Gamma-Hydroxybutyric Acid (GHB) and
Gamma-Butyrolactone (GBL) in Urine and
Blood Matrices
Joshua Z. Seither, BS*, 3645 Spring Valley Road, Akron, OH 44333; and
H.M. Kingston, PhD, and Timothy Fahrenholz, BS, Duquesne University,
700 Forbes Avenue, Pittsburgh, PA 15219
After attending this presentation, attendees will learn how Speciated
Isotope Dilution Mass Spectrometry (SIDMS) can be applied to the
forensic community. This presentation will provide an example of GHB
and GBL to show how this method can be applied.
This presentation will impact the forensic science community by
showing the community a quicker and more accurate way to quantitate
drugs with the example of GHB and GBL.
There are currently two major problems in the forensic science
community: scrutiny of analytical methods and a rapidly growing
backlog of samples. An accurate, rapid and simultaneous measurement
of GHB and GBL in urine and blood was developed to combat these
issues. This legally defensible method for analyzing both gammahydroxybutyric
acid (GHB) and gamma-butyrolactone (GBL)
simultaneously uses speciated isotope dilution mass spectrometry
(SIDMS). Current methods use gas chromatography mass spectrometry
(GC/MS) and are not able to quantitate both GHB and GBL
simultaneously; therefore, multiple extractions are required in order to
quantitatively analyze GHB and GBL. To perform SIDMS, deuterium
labeled GHB and carbon labeled GBL were utilized to spike the samples
for quantitation. Once the naturally occurring analyte is spiked with the
isotopically enriched analyte, SIDMS can account for any interconversion
that occurs between GHB and GBL during sample
preparation or analysis. After spiking the samples, a mixed-mode
(phenyl and propyl sulfonic acid) solid phase extraction column was
used for the filtration extraction of GHB and GBL from urine and blood
samples. Mass spectrometry studies were done using electrospray
ionization. Method validation was completed with triplicate sample
preparation and analyses (n =9) with a known concentration of GHB and
GBL in standardized urine and blood. Significant values of GHB and
GBL were chosen based on previous studies completed in the literature.
Concentration values of 5 ppm, 10 ppm, 200 ppm, and 400 ppm were
used. Endogenous levels of GHB average below 10 ppm. Some studies
have reported endogenous cutoff levels of GHB should be 6 ppm in urine
to avoid false negatives. GHB overdoses were reported at an average of
200 ppm and have been seen as high as 400 ppm. The experimental
values and the standard values were in agreement with the 95%
confidence interval. By using SIDMS, inter-conversions between GHB
and GBL can be accounted for and the correct quantification of both
analytes can be made. Temperature and pH levels were varied to
stimulate conversion between the two analytes, GHB and GBL. The
inter-conversion was accounted for in the SIDMS calculation, which
demonstrates the benefit for the use of this method in the forensic science
community. Calculations were made to account for the inter-conversion,
which demonstrate the use of the SIDMS method for drug quantitation.
* Presenting Author
This method can help forensic scientists by providing a procedure that is
legally defensible and quicker than other traditional methods of
analyzing GHB and GBL. This method can be beneficial to the forensic
science community.
Quantitation, SIDMS, GHB
K18 Validating Immunoassay ELISA Kits
to Detect Eighteen Benzodiazepines at
Low Levels
Danielle C. Mata, MS*, 320 North Flower Street, Santa Ana, CA 92703
After attending the presentation, attendees will understand the
process of validating an immunoassay benzodiazepine ELISA kit for low
concentration of drugs.
This presentation will impact the forensic science community by
demonstrating how changing the experimental parameters for an ELISA
kit will allow you to detect more drugs at needed concentrations.
Reports have shown that 30-40% of drivers take benzodiazepines
and that the use of these drugs could have impairing effects. The Orange
County Crime Lab (OCCL) recently validated an immunoassay
benzodiazepine ELISA screen to detect the 22 benzodiazepines
confirmed by an LC/MS/MS method at similar detection levels. The
main benzodiazepines prevalent in casework are alprazolam, diazepam,
lorazepam, and clonazepam and detection limits of 2, 10, 4, and
3 ng/mL, respectively are required. The validation process addressed
limits of detection, blanks, sample volumes, possible interferences, and
saturation curves for all detected benzodiazepines. The validation
determined that Temazepam at 3 ng/mL is the best benzodiazepine to use
for the limit of detection, and allows the OCCL to detect 18 of the 22
benzodiazepines seen via LC/MS/MS. Only four benzodiazepine
metabolites yield false negative results when no other benzodiazepines
are present.
ELISA Validation, Benzodiazepines, Low Concentrations
K19 Detection of Acute Diazinon Exposure in
Postmortem Bone Samples
Nebile Daglioglu, PhD*, Cukurova University, School of Medicine,
Department of Forenisc Medicine, Adana, 01330, TURKEY; Ramazan
Akcan, MD, Dicle University, School of Medicine, Department of Forensic
Medicine, Merkez Ilce 21280, Diyarbakir, 21280, TURKEY; Mete K.
Gulmen, PhD, Cukurova University, School of Medicine, Department of
Forensic Medicine, Adana, 01330, TURKEY; and Fadile Yener, MS, and
Pinar Efeoglu, MS, Cukurova University, School of Medicine, Department
of Forensic Medicine, Balcali, Adana, 01330, TURKEY
After attending this presentation, attendees will understand the
techniques of analyzing the acute diazinon exposure in postmortem bone
samples in explaining the mechanism and cause of death.
This presentation will impact the forensic science community by
the mechanism of diazinon exposure in understanding the cause of death
and the importance of detecting it with the forensic toxicology
lab techniques.
Forensic toxicological analyses have traditionally focused on the
use of blood, body fluids, and certain organs in examinations of deaths
due to intoxication. However, in some situations, putrefaction and
contamination make proper sampling from tissues and blood impossible,
such as in exceedingly degraded exhumation cases. In these cases, bone
might be useful as an alternative specimen since it is a potential depot for
pesticides and other chemical agents.
This third study is focused on the use of alternative specimens
where putrefaction and contamination make proper sampling from
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