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An enhanced butyrylcholinesterase method to measure organophosphorus nerve agent exposure in humans

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Abstract

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 μL 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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Abbreviations

A:

Alanine

Ab:

Antibody

AChE:

Acetylcholinesterase

BChE:

Butyrylcholinesterase

ChE:

Cholinesterase

CLSI:

Clinical and Laboratory Standards Institute

E:

Glutamic acid

F:

Phenylalanine

G:

Glycine

GB:

Sarin

IMS:

Immunomagnetic separation

LC-MS/MS:

Liquid chromatography tandem mass spectrometry

LOD:

Limit of detection

LOQ:

Limit of quantitation

LRL:

Lowest reportable limit

MeP:

Methylphosphonic acid

MRM:

Multiple reaction monitoring

OP:

Organophosphorus

OPNA:

Organophosphorus nerve agents

QC:

Quality control

QCH:

Quality control high

QCL:

Quality control low

RSD:

Relative standard deviation

S:

Serine

TNO:

The Netherlands Organisation for Applied Scientific Research

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Acknowledgments

The authors would like to thank the members of the Analytical Response and Chemical Laboratory Response Network Laboratories for technical assistance and thoughtful discussion during the preparation of this manuscript. Specifically, the authors would like to thank Ms. Chariety Sapp of the Centers for Disease Control and Prevention’s Incident Response Laboratory for aliquoting and distributing the convenience set specimens and Ms. Karen Shields for thoughtful input to the manuscript.

Funding information

This work was funded by the Centers for Disease Control and Prevention, Defense Threat Reduction Agency, and Oak Ridge Institute for Science and Education.

Disclaimer

The findings and conclusions in this report are those of the authors and do not necessarily represent the views of the Centers for Disease Control and Prevention. Use of trade names is for identification only and does not imply endorsement by the Centers for Disease Control and Prevention, the Public Health Service, or the US Department of Health and Human Services.

Author information

Authors and Affiliations

  1. Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, 4770 Buford Highway, NE, Atlanta, GA, 30341, USA

    Brooke G. Pantazides, Melissa D. Carter, Brian S. Crow, Thomas A. Blake, Jerry D. Thomas & Rudolph C. Johnson

  2. Oak Ridge Institute for Science and Education Fellow at the Centers for Disease Control and Prevention, 4770 Buford Highway, NE, Atlanta, GA, 30341, USA

    Caroline M. Watson

  3. Battelle, 2987 Clairmont Road, Atlanta, GA, 30329, USA

    Jonas W. Perez

Authors
  1. Brooke G. Pantazides
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  2. Caroline M. Watson
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  3. Melissa D. Carter
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  4. Brian S. Crow
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  5. Jonas W. Perez
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  6. Thomas A. Blake
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  7. Jerry D. Thomas
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  8. Rudolph C. Johnson
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Corresponding author

Correspondence to Rudolph C. Johnson.

Additional information

Published in the topical collection Analysis of Chemicals Relevant to the Chemical Weapons Convention with guest editors Marc-Michael Blum and R. V. S. Murty Mamidanna.

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Pantazides, B.G., Watson, C.M., Carter, M.D. et al. An enhanced butyrylcholinesterase method to measure organophosphorus nerve agent exposure in humans. Anal Bioanal Chem 406, 5187–5194 (2014). https://doi.org/10.1007/s00216-014-7718-7

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  • DOI: https://doi.org/10.1007/s00216-014-7718-7

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