title: Dichlorvos, chlorpyrifos oxon and Aldicarb adducts of butyrylcholinesterase, detected by mass spectrometry in human plasma following deliberate overdose.
abstract: The goal of this study was to develop a method to detect pesticide adducts in tryptic digests of butyrylcholinesterase in human plasma from patients poisoned by pesticides. Adducts to butyrylcholinesterase in human serum may serve as biomarkers of pesticide exposure because organophosphorus and carbamate pesticides make a covalent bond with the active site serine of butyrylcholinesterase. Serum samples from five attempted suicides (with dichlorvos, Aldicarb, Baygon and an unknown pesticide) and from one patient who accidentally inhaled dichlorvos were analyzed. Butyrylcholinesterase was purified from 2 ml serum by ion exchange chromatography at pH 4, followed by procainamide affinity chromatography at pH 7. The purified butyrylcholinesterase was denatured, digested with trypsin and the modified peptide isolated by HPLC. The purified peptide was analyzed by multiple reaction monitoring in a QTRAP 4000 mass spectrometer. This method successfully identified the pesticide-adducted butyrylcholinesterase peptide in four patients whose butyrylcholinesterase was inhibited 60-84%, but not in two patients whose inhibition levels were 8 and 22%. It is expected that low inhibition levels will require analysis of larger serum plasma volumes. In conclusion, a mass spectrometry method for identification of exposure to live toxic pesticides has been developed, based on identification of pesticide adducts on the active site serine of human butyrylcholinesterase.
authors.affiliation: Eppley Institute, University of Nebraska Medical Center, Omaha, NE 68198-5950, USA.
authors.forename: Bin
authors.initials: B
authors.lastname: Li
authors.forename: Ivan
authors.initials: I
authors.lastname: Ricordel
authors.forename: Lawrence M
authors.initials: LM
authors.lastname: Schopfer
authors.forename: Frédéric
authors.initials: F
authors.lastname: Baud
authors.forename: Bruno
authors.initials: B
authors.lastname: Mégarbane
authors.forename: Patrick
authors.initials: P
authors.lastname: Masson
authors.forename: Oksana
authors.initials: O
authors.lastname: Lockridge
mesh_terms: D000448:Aldicarb / Q000097:blood* / Q000378:metabolism / Q000506:poisoning; D001665:Binding Sites; D002091:Butyrylcholinesterase / Q000097:blood* / Q000378:metabolism; D004390:Chlorpyrifos / Q000031:analogs & derivatives* / Q000097:blood / Q000378:metabolism / Q000506:poisoning; D002800:Cholinesterase Inhibitors / Q000097:blood* / Q000378:metabolism / Q000506:poisoning; D002851:Chromatography, High Pressure Liquid; D004006:Dichlorvos / Q000097:blood* / Q000378:metabolism / Q000506:poisoning; D006801:Humans; D007306:Insecticides / Q000097:blood* / Q000378:metabolism / Q000506:poisoning; D011041:Poisoning / Q000097:blood; D011485:Protein Binding; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013406:Suicide, Attempted
chemical_list: D002800:Cholinesterase Inhibitors; D007306:Insecticides; C009618:O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphate; D004006:Dichlorvos; D000448:Aldicarb; D002091:Butyrylcholinesterase; D004390:Chlorpyrifos
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