Detection of sarin hydrolysis products from sarin-like organophosphorus agent-exposed human erythrocytes.
A sarin-like organophosphorus agent, [bis(isopropyl methyl)phosphonate; BIMP], was synthesized. This agent has the same phosphonate group as sarin and also has the same anti-acetylcholinesterase activity potency as sarin. The ID50 and LD50 values of BIMP in mice after intravenous injection were 3.9 nM and 0.8 mg/kg, respectively. The AChE activities of their red blood cells and brains were dose-dependently reduced by intravenous BIMP. After preparation of experimental BIMP-exposed human red blood cells, BIMP-bound acetylcholinesterase (AChE) was solubilized from erythrocyte membranes, purified by immunoaffinity chromatography, digested with trypsin, and the sarin hydrolysis products bound to AChE were released by alkaline phosphatase digestion. The digested sarin hydrolysis products were subjected to trimethylsilyl (TMS) derivatization and detected by gas chromatography-mass spectrometry. Isopropyl methylphosphonic- and methylphosphonic acids, which are the sarin hydrolysis products, were detected in experimental BIMP-exposed human red blood cells. This new method, which enables sarin's hydrolysis products to be detected in BIMP-exposed erythrocytes, is a useful tool for studying sarin-poisoning victims.
Nagao
M
M
Department of Forensic Medicine, Faculty of Medicine, University of Tokyo, Hongo, Japan.
Takatori
T
T
Matsuda
Y
Y
Nakajima
M
M
Niijima
H
H
Iwase
H
H
Iwadate
K
K
Amano
T
T
D000110:Acetylcholinesterase / Q000097:blood; D000818:Animals; D002619:Chemical Warfare Agents / Q000378:metabolism* / Q000506:poisoning; D002800:Cholinesterase Inhibitors / Q000097:blood* / Q000138:chemical synthesis / Q000506:poisoning; D004164:Diphosphonates / Q000097:blood* / Q000138:chemical synthesis / Q000506:poisoning; D004912:Erythrocytes / Q000201:enzymology*; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D006868:Hydrolysis; D007928:Lethal Dose 50; D051379:Mice; D008807:Mice, Inbred BALB C; D009943:Organophosphorus Compounds / Q000097:blood; D012524:Sarin / Q000378:metabolism* / Q000506:poisoning
D002619:Chemical Warfare Agents; D002800:Cholinesterase Inhibitors; D004164:Diphosphonates; D009943:Organophosphorus Compounds; C109992:bis(isopropyl methyl)phosphonate; C032627:methylphosphonic acid; D012524:Sarin; D000110:Acetylcholinesterase