title: Chromatographic analysis of toxic phosphylated oximes (POX): a brief overview.
abstract: Poisoning with organophosphorus compounds (OP), e.g. pesticides and nerve agents, causes inhibition of acetylcholinesterase (AChE) by phosphylation of the active site serine residue. Consequently, accumulation of stimulating acetylcholine in the synaptic cleft induces cholinergic crisis which ultimately may lead to death. For standard causal therapy, enzyme reactivators are administered representing oxime derivatives of quarternary pyridinium compounds, e.g. pralidoxime (2-PAM), obidoxime and HI 6. The mechanism of action includes removal of the phosphyl moiety by a nucleophilic attack of the oximate molecule substituting the enzyme and forming a phosphylated oxime (POX). POX is produced in stoichiometric amounts of reactivated enzyme and exhibits a significantly enhanced toxicity (inhibition rate constant) when compared to the parent OP. However, stability of POX under physiological conditions appears to be highly limited. Nevertheless, the presence of POX reveals a potential critical issue for both therapeutic efficacy in vivo and pharmacokinetic and pharmacodynamic (PK-PD) modelling based on cholinesterase activity data. Detailed characterization represents an important need for elaboration of the entire oxime pharmacology.Nevertheless, reports on POX toxicity and analysis are quite rare and may therefore be indicative of the challenge of POX analysis. This review provides a concise overview of chromatographic approaches applied to POX separation. Chromatography represents the key technology for POX purification and quantification in kinetic in vitro studies using buffers and biological fluids. Applications based on reversed-phase chromatography (RPC), ion pair chromatography (IPC) and an affinity approach as well as thin layer chromatography (TLC) are discussed and novel applications and data are presented.
authors.affiliation: Bundeswehr Institute of Pharmacology and Toxicology, Munich, Germany. HaraldJohn@bundeswehr.org
authors.forename: Christian
authors.initials: C
authors.lastname: Becker
authors.forename: Franz
authors.initials: F
authors.lastname: Worek
authors.forename: Harald
authors.initials: H
authors.lastname: John
mesh_terms: D000818:Animals; D002619:Chemical Warfare Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification*; D002845:Chromatography / Q000379:methods*; D006801:Humans; D009943:Organophosphorus Compounds / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification*; D010091:Oximes / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification*; D010575:Pesticides / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D002619:Chemical Warfare Agents; D009943:Organophosphorus Compounds; D010091:Oximes; D010575:Pesticides