ArticleForensic toxicology2026
Post-mortem degradation of flunitrazepam by aldehyde oxidase 1 and hemoglobin in blood.
Article in Forensic toxicology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
purposeFlunitrazepam is a widely used hypnotic agent detected in many fatal intoxication and suicide cases. However, its concentration in blood during autopsy differs from that at the time of death; it is either undetected or detected at lower concentrations than expected in post-mortem blood. In this study, we evaluated interactions between flunitrazepam and human blood using liquid chromatography coupled with quadrupole-Orbitrap mass spectrometry (LC-Q-Orbitrap-MS).
methodsWe measured flunitrazepam, 7-aminoflunitrazepam via reduction by aldehyde oxidase 1 (AOX1), and the degradation product of 7-aminoflunitrazepam via the Fenton reaction by hemoglobin (Hb) in human blood. In addition, we also detected flunitrazepam, 7-aminoflunitrazepam, and their degradation products in the blood of individuals who died from intentional ingestion of flunitrazepam.
resultsWe found that flunitrazepam was reduced to 7-aminoflunitrazepam by AOX1 in blood, and 7-aminoflunitrazepam was oxidized by Hb via the Fenton reaction. The mass spectra obtained by LC-Q-Orbitrap-MS revealed the formation of 7-amino-5-(2-fluorocyclohexa-1,3-dien-1-yl)-3-hydroxy-1-methyl-1,3-dihydro-2H-benzo[e][1,4]diazepin-2-one (7-amino-3-hydroxyflunitrazepam) in the Hb/H
conclusion7-Amino-3-hydroxyflunitrazepam is the degradation product of flunitrazepam at various temperatures. The degradation is brought about by post-mortem changes via AOX1-mediated reduction and Hb-mediated Fenton reaction. Thus, 7-amino-3-hydroxyflunitrazepam may be a valuable biomarker for correcting blood flunitrazepam concentration at the time of death in clinical cases.
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