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Published:
Journal of Analytical Toxicology,
ISSN 0146-4760,
Volume 32, Number 7, September,
pp.457-469
?Simultaneous
Detection and Quantification of Amphetamines, Diazepam and its
Metabolites, Cocaine and its Metabolites, and Opiates in Hair by
LC–ESI-MS–MS Using a Single Extraction Method
?Eleanor I. Miller, Fiona M. Wylie, and John S. Oliver
Forensic Medicine and Science, University of Glasgow, University
Place, Glasgow, G12 8QQ, Scotland
?A liquid chromatography–tandem mass spectrometry
method was developed and validated for the simultaneous identification
and quantification of amphetamines, diazepam and its metabolites,
cocaine and its metabolites, and opiates from hair using a single
extraction method. As part of the method development, Gemini
C18, Synergi Hydro RP, and Zorbax Stablebond-Phenyl LC columns
were tested with three different mobile phases. Analyte recovery
and limit of detection were evaluated for two different solid-phase
extraction methods that used Bond Elut Certify™ and Clean
Screen® cartridges. Phosphate buffer (pH 5.0) was chosen
as the optimum hair incubation medium because of the high stability
of cocaine and 6-monoacetylmorphine using this method and faster
sample preparation. The optimized method was fully validated.
Linearity was established over the concentration range 0.2–10
ng/mg hair, and the correlation coefficients were all greater
than 0.99. Total extraction recoveries were greater than 76%,
detection limits were between 0.02 and 0.09 ng/mg, and the intra-
and interday imprecisions were generally less than 20% in spiked
hair. The intra- and interbatch imprecision of the method for
a pooled authentic hair sample ranged from 1.4 to 23.4% relative
standard deviation (RSD) and 8.3 to 25.4% RSD, respectively,
for representative analytes from the different drug groups. The
percent matrix effect ranged from 63.5 to 135.6%, with most analytes
demonstrating ion suppression. Sixteen postmortem samples collected
from suspected drug-related deaths were analyzed for the 17 drugs
of abuse and metabolites included in the method. The method was
sufficiently sensitive and specific for the analysis of drugs
and metabolites in postmortem hair samples. There is scope for
the inclusion of other target drugs and metabolites in the method.
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