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Published:
Journal of Analytical Toxicology,
ISSN 0146-4760,
Volume 31, Number 2, March 2007,
pp.69-74
A Rapid Cation-Exchange HPLC Method for Detection and
Quantification of Pyridinium Oximes in Plasma and Tissue
Harry Singh, Deborah Moorad-Doctor, Ruthie H. Ratcliffe,
Katie Wachtel, Andres Castillo, and Gregory E. Garcia
Department of Biochemical Pharmacology, Division of Biochemistry, Walter Reed
Army Institute of Research, Silver Spring, Maryland 20910
A rapid and sensitive assay for pyridinium oximes in plasma and
tissue was developed. The method was suitable for the analysis of mono- and
di-pyridinium oximes and utilizes ultrafiltration followed by cation-exchange
high-performance liquid chromatography with UV detection. The assay was originally
developed for the measurement of the oxime MMB-4 in plasma for which the lower
limit of detection was 0.0005 µg and the limit of quantitation was 0.001
to 2.5 µg. The assay required as little as 50 µL of whole blood
or 30 µL of tissue homogenate, and it was used for a pharmacokinetic study
from a single intramuscular injection of MMB-4 (dichloride or dimethylsulfonate
salt) in the guinea pig. Both salts were found to have similar pharmacokinetic
properties in the plasma with a T? of about 34 to 42 min and the area-under-the-curve
values increased dose dependently. MMB-4 tissue concentrations were much lower
than the plasma. The tissue levels peaked at 5–20 min depending on the
tissue. A rank of concentration was diaphragm > heart > thigh muscle.
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