Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 406, Issue 21, Pages 5187-5194Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-014-7718-7
Keywords
Organophosphorus nerve agent; Butyrylcholinesterase; Cholinesterase inhibitors; Protein adduct; Immunomagnetic separation
Funding
- Centers for Disease Control and Prevention, Defense Threat Reduction Agency
- Oak Ridge Institute for Science and Education
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Organophosphorus nerve agent (OPNA) adducts to butyrylcholinesterase (BChE) can be used to confirm exposure in humans. A highly accurate method to detect G- and V-series OPNA adducts to BChE in 75 mu L of filtered blood, serum, or plasma has been developed using immunomagnetic separation (IMS) coupled with liquid chromatography tandem mass spectrometry (LC-MS/MS). The reported IMS method captures > 88 % of the BChE in a specimen and corrects for matrix effects on peptide calibrators. The optimized method has been used to quantify baseline BChE levels (unadducted and OPNA-adducted) in a matched-set of serum, plasma, and whole blood (later processed in-house for plasma content) from 192 unexposed individuals to determine the interchangeability of the tested matrices. The results of these measurements demonstrate the ability to accurately measure BChE regardless of the format of the blood specimen received. Criteria for accepting or denying specimens were established through a series of sample stability and processing experiments. The results of these efforts are an optimized and rugged method that is transferrable to other laboratories and an increased understanding of the BChE biomarker in matrix.
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