LC-MS-MS analysis of 2,4-dinitrophenol and its phase I and II metabolites in a case of fatal poisoning

Abstract

A liquid chromatography-tandem mass spectrometry (LC-MS-MS) analysis of biological fluids (blood, urine, gastric content, and bile) collected at autopsy in a case of suspected 2,4-dinitrophenol (DNP) fatal poisoning allowed the determination of DNP and its known metabolites (2-amino-4-nitrophenol and 2-nitro-4-aminophenol). The tentative identification of three conjugated metabolites (DNP glucuronide, DNP sulfate, and 2-amino-4-nitrophenol glucuronide) could be made on the basis of their pseudomolecular ion, isotopic and fragmentation patterns, and retention characteristics. Another DNP metabolite reported in the literature, 2,4-diaminophenol, was not detected in the samples. Postmortem blood concentrations were 48.4 mg/L for DNP and 1.2 mg/L for 2-amino-4-nitrophenol. Gas chromatography-MS screening and quantification in postmortem blood revealed the presence of toxic concentrations of citalopram and its desmethylated metabolite (0.58 and 0.40 mg/L, respectively) and therapeutic or lower than therapeutic levels of olanzapine (0.04 mg/L), desalkylflurazepam (0.02 mg/L), and nordazepam (0.01 mg/L). Based on LC-MS-MS results and on available literature data on DNP poisonings, it was concluded that DNP poisoning played a contributing role, together with citalopram, in the cause of death.