SOFT 2004 Meeting Abstracts - Society of Forensic Toxicologists

P48
DETERMINATlON OF FENTANYL IN POSTMORTEM BLOOD SAMPLES BY LIQUID
CHROMA TOGRAPHY TANDEM MASS SPECTROMETRY
Vera Coopman l ', Dirk. Van Varenbergh 2 , Karen Pien 2 and Jan Cordonnier l
lDepartment of Analytical Toxicology, Chemiphar N.V., Lieven Bauwensstraat 4, Bruges, Belgium.
2Department of Forensic Medicine, Free Brussels University, Brussels, Belgium.
Fentanyl is a potent synthetic narcotic analgesic administrated in the form of a transdermal patch for the
management of chronic pain. The authors present a liquid-liquid extraction procedure and a liquid
chromatography-tandem mass spectrometry method (LC-MS-MS) for the quantitation of fentanyl in
postmortem blood samples .. The method was applied in a case of acute intoxication due to excessive
administrated Durogesic® transdermal patch.
To a 1.0 mL aliquot of blood sample, 50 flL of internal standard solution fentanyl-ds (1 flgJmL) and 1 mL
1M K 2 C0 3 was added. Extraction was performed with 7 mL of a mixture of n-hexane:ethylacetate (7:3,
VN). After vortex mixing and centrifugation, the upper organic layer was evaporated and reconstituted in
1.0 mL of initial mobile phase. The LC-MS-MS analysis were performed using an Alliance® 2695 system
and a Quatro Micro mass spectrometer (Waters Milford, USA). Chromatography was achieved using a
Xterra® MS C18 column 2.1 mm x 100 mm with a 3.5 flm particle size (Waters Milford, USA). A gradient
elution was used with 0.15% formic acid and 0.15% formic acid in acetonitrile. The chromatographic
duration was 30 minutes. The flow rate was 0.20 mLimin and the column oven temperature was set at
35°C. A 20 flL aliquot of the sample was injected. Multiple reaction monitoring (MRM) with positive ion
dectection (ES+) was used for selective detection of fentanyl and fentanyl-ds. The precursor ion and two
fragment ions were selected as quantifier and qualifier, for fentanyl mlz 336.8 and fragments at m/z 187.95
and 104.4, for fentanyl-ds mlz 341.9 and fragments at m/z 188.0 and 104.5. Calibration was performed by
addition of standard solutions to fentanyl-free blood prior to extraction. Precision, accuracy and recovery
experiments were carried out. The liquid-liquid extraction procedure in combination with LC-MS-MS
proved to be a good alternative compared with the more labor-intensive solid-phase extractions previously
described.
The analytical procedure was applied in a forensic case: a 78-year-old woman was found death in bed,
lying on het back. The external examination revealed 10 Durogesic® transdermal patches (fentanyl 100
flglh) on the abdomen. An autopsy was carried out 2 days later. Multiple samples were taken for
toxicological examination. The following fentanyl concentrations were found; 28.6 flglL (right) and 28.2
fJg/L (left) in subclavian blood, 21.3 fJglL (right) and 20.9 fJg/L (left) in femoral blood, 37.6 fJglL (right)
and 33.9 flg/L (left) in ventricular blood. The comprehensive systematic toxicological analysis did not
reveal the presence of an other substance which contributed to the death of the woman. We concluded that
death was caused by intoxication with fentanyl. The manner was presumed to be suicide.
Keywords: fentanyl, LC-MS-MS, fatality
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