SOFT 2004 Meeting Abstracts - Society of Forensic Toxicologists

A78
LCIMSIMS DETERMINATION OF LSD, ISO-LSD, AND THE MAIN METABOLITE 2-0XO-3­
HYDROXY-LSD IN WHOLE BLOOD AND URINE
Sys Stybe Johansen* and Jytle Lundsby Jensen
Department of Forensic Chemistry, Institute of Forensic Medicine, University of Copenhagen,
Frederik V's vej 11, DK-2100 Copenhagen OE, Denmark. Phone +45-35326241. Fax: +45-35326085. E­
mail: SSJ@forensic.ku.dk
Aim: To develop simple LC/MSIMS applications that identify, confirm and quantify rare potent drugs in
forensic cases. An application with the very potent hallucinogenic drug LSD is shown.
Method: A liquid chromatography mass spectrometric (LCIMSIMS) method has been developed for the
determination of LSD in whole blood and urine. Furthermore, determination of the common LSD impurity
iso-LSD (inactive) and the main metabolite 2-oxo-3-hydroxy-LSD were included. The procedure involved
a simple liquid-liquid extraction of I g sample containing the analyte and LSD-D3 (internal standard) with
butyl acetate at pH 9.8. After centrifugation, the organic fraction was removed and evaporated to dryness at
40°C and reconstituted in 100 III mobile phase. A gradient LC system (HP 1100 system, Agilent tech.) with
a Zorbax SB CI8 (30 x 2.lmm, 3.5 Ilm) was used to separate the analytes within 10 min. Identification,
confirmation and quantification were done by positive electrospray ionisation with a triple quadrupole mass
spectrometer (Quattro micro, Waters) operating in multiple reaction monitoring (MRM) mode. For each
analyte two MRM's were set up, one for quantification and one as qualifier using one precursor ion and
two product ions per analyte. The ratio between the responses of the two MRM's was used for
identification purposes along with the retention time.
Results: The curves of extracted whole blood standards were linear over a working range of 0.01 to 5.0
Ilg/kg for LSD and iso-LSD of both transitions. The limit of quantification (LOQ) of LSD and iso-LSD was
O.ot Ilglkg in whole blood, while LOQ of 2-oxo-3-hydroxy-LSD was 0.5 Ilglkg. The repeatability
expressed by relative standard deviation (RSD) was better than 10% and the relative accuracy was between
92-99% for LSD and 89-108% for iso-LSD, respectively. The ratio was determined for each analyte and a
RSD of max. 10% was confirmed.
The method was applied for a case investigation involving a 26-year-old man who admitted to have been
abusing LSD. Blood concentrations of LSD and iso-LSD were determined to 0.27 Ilglkg and 0.44 Ilglkg. 2­
oxo-3-hydroxy-LSD was detected in the urine sample and confirmed the abuse of LSD. The case illustrated
the importance of analyte separation before MRM detection of a sample. The chromatographic separation
of LSD and iso-LSD were of importance because the diastereoisomers have identical fragmentation paths
leading to fragments ofequal m/z ratio.
Conclusion: The method was proved to be suitable for forensic cases being simple, reproducible and
selective. However, it was observed that monitoring typical fragments of LSD without chromatographic
separation is inadequate. After the development of this LCIMSIMS method, the initial screening by RIA
(radioimmunoassay) for LSD was cancelled because it was too time-consuming and expensive considering
the low number of investigations/cases per year.
Keywords: LSD, Whole blood, LC/MS/MS
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