WRAIR Protocols for Soldier Status and Readiness to Organophosphate Exposure: Unprocessed Whole Blood Cholinesterase and Pyridostigmine Bromide Quantification
WALTER REED ARMY INST OF RESEARCH SILVER SPRING MD
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Exposure to nerve agents, OPs, pesticides, anesthetics, terrorists chemical agents, and drugs of abuse such as cocaine, heroin, and some neuro-degenerative disease states selectively reduces AChE andor BChE activity. Since urban terrorism is on the rise, Federal, State, and local authorities need a reliable, fast, inexpensive method for confirming such an assault to initiate appropriate containment, decontamination, and treatment measures. Thus, we developed a semi-automated medical diagnostic microplate procedure capable of screening unprocessed whole blood samples for the concentrations of AChE and BChE patent pending, WRAIR to overcome limitations of current methods. We applied our technique to whole blood in man, non-human primates, and rodents. In addition, pyridostigmine bromide PB treatment is the only prophylactic treatment for troops who expect to be exposed to OPs. Thus, we developed a sensitive HPLC technique to quantify PB in human blood and also in Rhesus and rodent blood and tissue. The technique uses solid phase extraction, lyophilization for concentration, and HPLC of the reconstituted samples followed by strong-cation exchange chromatography via isocratic elution. These techniques will provide the soldier, field medic, hospital, company, or civilian organization with the ability to rapidly determine exposure to OPs and aid in treatment while the HPLC assay will aid in verifying troop compliance when required for PB prophylaxis against OPs.
- Medicine and Medical Research
- Chemical, Biological and Radiological Warfare