Analytical and Bioanalytical Chemistry

, Volume 406, Issue 21, pp 5187–5194 | Cite as

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

  • Brooke G. Pantazides
  • Caroline M. Watson
  • Melissa D. Carter
  • Brian S. Crow
  • Jonas W. Perez
  • Thomas A. Blake
  • Jerry D. Thomas
  • Rudolph C. JohnsonEmail author
Research Paper
Part of the following topical collections:
  1. Analysis of Chemicals Relevant to the Chemical Weapons Convention


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.


Organophosphorus nerve agent Butyrylcholinesterase Cholinesterase inhibitors Protein adduct Immunomagnetic separation 













Clinical and Laboratory Standards Institute


Glutamic acid








Immunomagnetic separation


Liquid chromatography tandem mass spectrometry


Limit of detection


Limit of quantitation


Lowest reportable limit


Methylphosphonic acid


Multiple reaction monitoring




Organophosphorus nerve agents


Quality control


Quality control high


Quality control low


Relative standard deviation




The Netherlands Organisation for Applied Scientific Research



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.


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.


  1. 1.
    Convention of the prohibition of the development, production, stockpiling, and use of chemical weapons and their destruction (2005) Organisation for the Prohibition of Chemical Weapons, Netherlands. Assessed 16 August 2013
  2. 2.
    Nicolet Y, Lockridge O, Masson P, Fontecilla-Camps JC, Nachon F (2003) J Biol Chem 278:41141–41147CrossRefGoogle Scholar
  3. 3.
    Grob D, Harvey JC (1957) J Clin Invest 37:350–368CrossRefGoogle Scholar
  4. 4.
    Nagao M, Takatori T, Matsuda Y, Nakajima M, Iwase H, Iwadate K (1997) Toxicol Appl Pharm 144:198–203CrossRefGoogle Scholar
  5. 5.
    Black RM, Read RW (2007) Toxin Rev 26:275–298CrossRefGoogle Scholar
  6. 6.
    Report on the alleged use of chemical weapons in the Ghouta Area of Damascus on 21 August 2013 (2013) Council on Foreign Relations, Netherlands. slideshow/Secretary_General_Report_of_CW_Investigation.pdf. Accessed 17 Sep 2013
  7. 7.
    Ellman G, Courtney D, Andres V, Featherstone R (1961) Biochem Pharm 7:88–95CrossRefGoogle Scholar
  8. 8.
    Worek F, Mast U, Kidelen D, Diepold C, Eyer P (1999) Clin Chim Acta 288:73–90CrossRefGoogle Scholar
  9. 9.
    Gallo MA, Lawryk NJ (1991) In: Hayes WJ Jr, Laws ER Jr (eds) Handbook of Pesticide Toxicology. Acedemic Press, New YorkGoogle Scholar
  10. 10.
    Noort D, Benschop HP, Black RM (2002) Toxicol Appl Pharm 184:116–126CrossRefGoogle Scholar
  11. 11.
    Swaim LL, Johnson RC, Zhou Y, Sandlin C, Barr JR (2008) J Anal Toxicol 32:774–777CrossRefGoogle Scholar
  12. 12.
    Adams TK, Capacio BR, Smith JR, Whalley CE, Korte WD (2005) Drug Chem Toxicol 27:77–91CrossRefGoogle Scholar
  13. 13.
    Li B, Ricordel I, Schopfer LM, Baud F, Mégarbane B, Nachon F, Masson P, Lockridge O (2010) Toxicol Sci 116:23–31CrossRefGoogle Scholar
  14. 14.
    Black RM, Noort D (2005) In: Mesilaakso M (ed) Chemical weapons convention analysis: sample collection, preparation, and analytical methods. John Wiley & Sons, EnglandGoogle Scholar
  15. 15.
    Riches J, Morton I, Read RW, Black RM (2005) J Chromatogr B 816:251–258CrossRefGoogle Scholar
  16. 16.
    Degenhardt C, Pleijsier K, Van der Schans MJ, Langenberg JP, Preston KE, Solano MI, Maggio VL, Barr JR (2004) J Anal Toxicol 28:364–371CrossRefGoogle Scholar
  17. 17.
    Sporty JLS, Lemire SW, Jakubowski EM, Renner JA, Evans RA, Williams RF, Schmidt JG, Van der Schans MJ, Noort D, Johnson RC (2010) Anal Chem 82:6593–6600CrossRefGoogle Scholar
  18. 18.
    Williams NH, Harrison JM, Read RW, Black RM (2007) Arch Toxicol 81:627–639CrossRefGoogle Scholar
  19. 19.
    Li B, Nachon F, Froment MT, Verdier L, Debouzy JC, Brasme B, Gillon E, Schopfer LM, Lockridge O, Masson P (2008) Chem Res Toxicol 21:421–431CrossRefGoogle Scholar
  20. 20.
    Nigg HN, Knaak JB (2000) Rev Environ Contam Toxicol 163:29–111Google Scholar
  21. 21.
    Black RM (2010) J Chromatogr B 878:1207–1215CrossRefGoogle Scholar
  22. 22.
    Aurbek N, Thiermann H, Eyer F, Eyer P, Worek F (2009) Toxicol 259:133–139CrossRefGoogle Scholar
  23. 23.
    Fidder A, Hulst AG, Noort D, de Ruiter R, van der Schans MJ, Benschop HP, Langenberg JP (2002) Chem Res Toxicol 15:582–590CrossRefGoogle Scholar
  24. 24.
    Knaack JS, Zhou Y, Abney CW, Jacob JT, Prezioso SM, Hardy K, Lemire SW, Thomas J, Johnson RC (2012) Anal Chem 84:9470–9477CrossRefGoogle Scholar
  25. 25.
    Taylor JK (1987) Quality assurance of chemical measurements. Lewis Publishers, MichiganGoogle Scholar
  26. 26.
    Westgard JO, Barry PL, Hunt MR (1981) Clin Chem 27:493–508Google Scholar
  27. 27.
    Caudill SP, Schleicher RL, Pirkle JL (2008) Stat Med 27:4094–4106CrossRefGoogle Scholar
  28. 28.
    Carter MD, Crow BS, Pantazides BG, Watson CM, DeCastro BR, Thomas JD, Blake TA, Johnson RC (2013) J Biomol Screen. doi: 10.1177/1087057113497799 Google Scholar
  29. 29.
    Vesper H, Emons H, Gnezda M, Jain CP, Miller WG, Rej R, Schumann G, Tate J, Thienpont L, Vaks JE (2010) Characterization and qualification of commutable reference materials for laboratory medicine; approved guidelines (C53-A). Clinical and Laboratory Standards Institute, PAGoogle Scholar
  30. 30.
    Carter MD, Crow BS, Pantazides BG, Watson CM, Thomas JD, Blake TA, Johnson RC (2013) Anal Chem. doi: 10.1021/ac4029714 Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg (outside the USA) 2014

Authors and Affiliations

  • Brooke G. Pantazides
    • 1
  • Caroline M. Watson
    • 2
  • Melissa D. Carter
    • 1
  • Brian S. Crow
    • 1
  • Jonas W. Perez
    • 3
  • Thomas A. Blake
    • 1
  • Jerry D. Thomas
    • 1
  • Rudolph C. Johnson
    • 1
    Email author
  1. 1.Division of Laboratory Sciences, National Center for Environmental HealthCenters for Disease Control and PreventionAtlantaUSA
  2. 2.Oak Ridge Institute for Science and Education Fellow at the Centers for Disease Control and PreventionAtlantaUSA
  3. 3.BattelleAtlantaUSA

Personalised recommendations