Journal des techniques analytiques et bioanalytiques

Abstrait

An Extraction Assay Analysis for Galanthamine in Guinea Pig Plasma and its Application to Nerve Agent Countermeasures

Wes E Steiner, Ioannis A Pikalov, Patrick T Williams, William A English and Corey J Hilmas

Galanthamine hydrobromide (GAL HBr), approved material for treatment of mild to moderate Alzheimer’s disease, is a centrally-acting reversible acetylcholinesterase inhibitor (AChEI) that is currently under evaluation as a therapeutic countermeasure against organophosphorus G- and V-Series nerve agents, which can induce rapid lethality in guinea pigs and humans. It has been shown that upon combination with atropine (ATR) and pyridine-2-aldoxime methochloride (2-PAM), a single dose of GAL administered before or soon after the acute exposure to a lethal dose of organophosphorus compounds can safely counteract toxicity in guinea pigs. To that end a new sample preparation extraction method analysis assay has been explored to enable future high-throughput, reproducible, and sensitive assays for the extraction of galanthamine in guinea pig plasma. Samples were prepared with diphenhydramine hydrochloride (DPH HCl) internal standard and recovered with 10 min liquid-liquid trichloromethane extractions. Samples were analyzed with a reversed phase liquid chromatographic column interfaced to a triple quadrupole mass spectrometer (LC/MS/MS) operating in the positive ion multiple reaction monitoring (MRM) Turbo Ionspray mode. Precursor to product ion (M+H)+ transitions of 288-to-213 m/z and 256-to-167 m/z for GAL and DPH were observed, respectively. Sample run times of 1.50 min were achieved.