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Published:
Journal of Analytical Toxicology,
ISSN 0146-4760,
Volume 30, Issue 3, April 2006,
pp. 196-201
More Reliable Brain Death Diagnosis
With Chromatographic Analysis of Midazolam, Diazepam, Thiopentone, and Active
Metabolites
Andreas Meinitzer, Winfried März, Harald Mangge,
and Gabriele Halwachs-Baumann
Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University
of Graz, Auenbruggerplatz 19, 8036 Graz, Austria
Brain death diagnosis may be confounded by centrally acting drugs.
The certainty of brain death diagnosis can be enhanced by demonstrating that
the concentrations of such drugs are well below the therapeutic range. A combined
high-performance liquid chromatography-based method was developed for the benzodiazepines
midazolam, 1-hydroxymidazolam, 1-hydroxymidazolam glucuronide, diazepam, and
nordiazepam and for the barbiturates thiopentone and pentobarbitone in serum
or plasma of critically ill patients. The lower limits of detection of the assays
for benzodiazepines and barbiturates were 2.5 ng/mL and 0.05 µg/mL. The
lower limits of the working ranges of these assays were set at 25 ng/mL and
0.5 µg/mL, respectively, and are below the lowest pharmacologically active
plasma concentrations of these drugs. Intra- and interday coefficients of variations
were less than 2.5% and 11.0% throughout, as determined with six replicates
(n = 6). These assays were accurate in that the relative difference between
actually measured and expected concentration never exceeded 12%. Utilization
of these assays will render the diagnosis of brain death more reliable.
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