Title
Quantitation of ortho-cresyl phosphate adducts to butyrylcholinesterase in human serum by immunomagnetic-UHPLC-MS/MS
Author
Johnson, D.
Carter, M.D.
Crow, B.S.
Isenberg, S.L.
Graham, L.A.
Erol, H.A.
Watson, C.M.
Pantazides, B.G.
van der Schans, M.J.
Langenberg, J.P.
Noort, D.
Blake, T.A.
Thomas, J.D.
Johnson, R.C.
Publication year
2015
Abstract
Tri-ortho-cresyl phosphate (ToCP) is an anti-wear, flame retardant additive used in industrial lubricants, hydraulic fluids and gasoline. The neurotoxic effects of ToCP arise from the liver-activated metabolite 2-(o-cresyl)-4H-1,3,2-benzodioxaphosphoran-2-one (cresyl saligenin phosphate or CBDP), which inhibits esterase enzymes including butyrylcholinesterase (BChE). Following BChE adduction, CBDP undergoes hydrolysis to form the aged adduct ortho-cresyl phosphoserine (oCP-BChE), thus providing a biomarker of CBDP exposure. Previous studies have identified ToCP in aircraft cabin and cockpit air, but assessing human exposure has been hampered by the lack of a laboratory assay to confirm exposure. This work presents the development of an immunomagnetic-UHPLC-MS/MS method for the quantitation of unadducted BChE and the long-term CBDP biomarker, oCP-BChE, in human serum. The method has a reportable range from 2.0ng/ml to 150ng/ml, which is consistent with the sensitivity of methods used to detect organophosphorus nerve agent protein adducts. The assay demonstrated high intraday and interday accuracy (≥85%) and precision (RSD≤15%) across the calibration range. The method was developed for future analyses of potential human exposure to CBDP. Analysis of human serum inhibited in vitro with CBDP demonstrated that the oCP-BChE adduct was stable for at least 72h at 4, 22 and 37 C. Compared to a previously reported assay, this method requires 75% less sample volume, reduces analysis time by a factor of 20 and demonstrates a threefold improvement in sensitivity. Published 2015. This article is a U.S. Government work and is in the public domain in the USA. © Published 2015. This article is a U.S. Government work and is in the public domain in the USA.
Subject
Observation, Weapon & Protection Systems
CBRN - CBRN Protection
TS - Technical Sciences
Butyrylcholinesterase
Dresyl saligenin phosphate
Jamaica ginger paralysis
Organophosphate-induced delayed neuropathy
Tri-ortho-cresyl-phosphate
Body fluids
Cabins (aircraft)
Calibration
Enzymes
Lubricants
Tribology
Butyrylcholinesterase
Esterase enzymes
Flame-retardant additives
Human exposures
Industrial lubricants
Jamaica
Neurotoxic effects
Organophosphate-induced delayed neuropathy
Cockpits (aircraft)
Cholinesterase
Tri ortho cresyl phosphate
Accuracy
Cholinesterase blood level
Controlled study
In vitro study
Limit of detection
Tandem mass spectrometry
Ultra performance liquid chromatography
Zingiber officinale
To reference this document use:
http://resolver.tudelft.nl/uuid:fd3c6c2a-1ae0-423a-83c6-d95a18622334
DOI
https://doi.org/10.1002/jms.3576
TNO identifier
527060
Publisher
John Wiley and Sons Ltd
ISSN
1076-5174
Source
Journal of Mass Spectrometry, 50 (4), 683-692
Document type
article