Improvements to the Fluoride Reactivation Method by Simple Organic Extraction for Retrospective Detection of Exposure to the Organophosphorus Nerve Agents in Human Plasma

Jin Young Lee^*, Yong Han Lee

Agency for Defense Development (ADD), Daejeon, Republic of Korea.

DOI: 10.4236/ijamsc.2014.23007                

Abstract

A rapid and simple organic extraction method for the determination of the chemical warfare agent, isopropyl methylphosphonofluoridate (sarin, GB) in human plasma has been developed using gas chromatography-tandem mass spectrometry (GC-MS/MS). In the course of method development, several organic solvents have been screened and chloroform show a low background and increase signal to noise ratio of GB among other organic solvents. Especially, the organic extraction method of reactivated GB from the human plasma has a 30% greater recovery yield than solid-phase ex-traction (SPE). This simple extraction method was successfully applied to the trace analysis of nerve agents in human plasma in the 3rd Organisation for the Prohibition of Chemical Weapons (OPCW) confidence building exercise on biomedical sample analysis.

Keywords

Chemical Warfare Agents, Isopropyl Methylphosphonofluoridate, Organic Extraction, Plasma

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Casida, J.E. and Quistad, G.B. (2004) Organophosphate Toxicology: Safety Aspects of Nonacetylcholinesterase Secondary Targets. Chemical Research in Toxicology, 17, 983-998. http://dx.doi.org/10.1021/tx0499259
[2]	Technical Secretariat of the Organization for Prohibition of Chemical Weapons (1997) Convention on the Prohibition of the Development, Production, Stockpiling and use of Chemical Weapons and their Destruction. The Hague. http://www.opcw.nl
[3]	Shih, M.L., Smith, J.R., McMonagle, J.D., Dolzine, T.W. and Gresham, V.C. (1991) Detection of Metabolites of Toxic Alkylmethylphosphonates in Biological Samples. Biological Mass Spectrometry, 20, 717-723. http://dx.doi.org/10.1002/bms.1200201111
[4]	Barr, J.R., Driskell, W.J., Aston, L.S. and Martinez, R.A. (2004) Quantitation of Metabolites of the Nerve Agents Sarin, Soman, Cyclohexylsarin, VX, and Russian VX in Human Urine Using Isotope-Dilution Gas Chromatography-Tandem Mass Spectrometry. Journal of Analytical Toxicology, 28, 372-378. http://dx.doi.org/10.1093/jat/28.5.372
[5]	Black, R.M., Clarke, R.J., Read, R.W. and Reid, M.T.J. (1994) Application of Gas Chromatography-Mass Spectrometry and Gas Chromatography-Tandem Mass Spectrometry to the Analysis of Chemical Warfare Samples, Found to Contain Residues of the Nerve Agent Sarin, Sulphur Mustard and Their Degradation Products. Journal of Chromatography A, 662, 301-321. http://dx.doi.org/10.1016/0021-9673(94)80518-0
[6]	Driskell, W.J., Shih, M., Needham, L.L. and Barr, D.B. (2002) Quantitation of Organophosphorus Nerve Agent Metabolites in Human Urine Using Isotope Dilution Gas Chromatography-Tandem Mass Spectrometry. Journal of Analytical Toxicology, 26, 6-10. http://dx.doi.org/10.1093/jat/26.1.6
[7]	Fredriksson, S.A., Hammarstrom, L.G., Henriksson, L. and Lakso, H.A. (1995) Trace Determination of Alkyl Methylphosphonic Acids in Environmental and Biological Samples Using Gas Chromatography/Negative-Ion Chemical Ionization Mass Spectrometry and Tandem Mass Spectrometry. Journal of Mass Spectrometry, 30, 1133-1143. http://dx.doi.org/10.1002/jms.1190300810
[8]	Black, R.M. and Read, R.W. (1997) Application of Liquid Chromatography-Atmospheric Pressure Chemical Ionisation Mass Spectrometry, and Tandem Mass Spectrometry, to the Analysis and Identification of Degradation Products of Chemical Warfare Agents. Journal of Chromatography A, 759, 79-92. http://dx.doi.org/10.1016/S0021-9673(96)00763-7
[9]	Black, R.M. and Read, R.W. (1998) Analysis of Degradation Products of Organophosphorus Chemical Warfare Agents and Related Compounds by Liquid Chromatography-Mass Spectrometry Using Electrospray and Atmospheric Pressure Chemical Ionization. Journal of Chromatography A, 794, 233-244. http://dx.doi.org/10.1016/S0021-9673(97)00611-0
[10]	Mawhinney, D.B., Hamelin, E.I., Fraser, R., Silva, S.S., Pavlopoulos, A.J. and Kobelski, R.J. (2007) The Determination of Organophosphonate Nerve Agent Metabolites in Human Urine by Hydrophilic Interaction Liquid Chromatography Tandem Mass Spectrometry. Journal of Chromatography B, 852, 235-243. http://dx.doi.org/10.1016/j.jchromb.2007.01.023
[11]	Noort, D., Hulst, A.G., Platenburg, D., Polhuijs, M. and Benschop, H.P. (1998) Quantitative Analysis of O-Isopropyl Methylphosphonic Acid in Serum Samples of Japanese Citizens Allegedly Exposed to Sarin: Estimation of Internal Dosage. Archives of Toxicology, 72, 671-675. http://dx.doi.org/10.1007/s002040050559
[12]	T?rnes, J.A. (1996) Identification of Some Alkyl Methylphosphonic Acids by Thermospray Tandem Mass Spectrometry. Rapid Communications in Mass Spectrometry, 10, 878-882. http://dx.doi.org/10.1002/(SICI)1097-0231(19960610)10:8%3C878::AID-RCM596%3E3.0.CO;2-B
[13]	Benschop, H. and De Jong, L. (2001) In: Somani, S. and Romano, J., Eds., Chemical Warfare Agents: Toxicity at Low Levels, CRC Press, Boca Raton, 25-81.
[14]	Fidder, A., Hulst, A.G., Noort, D., de Ruiter, R., van der Schans, M.J., Benschop, H.P. and Langenberg, J.P. (2002) Retrospective Detection of Exposure to Organophosphorus Anti-Cholinesterases:? Mass Spectrometric Analysis of Phosphylated Human Butyrylcholinesterase. Chemical Research in Toxicology, 15, 582-590. http://dx.doi.org/10.1021/tx0101806
[15]	Jakubowski, E.M., McGuire, J.M., Evans, R.A., Edwards, J.L., Hulet, S.W., Benton, B.J., Forster, J.S., Burnett, D.C., Muse, W.T., Matson, K., Crouse, C.L., Mioduszewski, R.J. and Thomson, S.A. (2004) Quantitation of Fluoride Ion Released Sarin in Red Blood Cell Samples by Gas Chromatography-Chemical Ionization Mass Spectrometry Using Isotope Dilution and Large-Volume Injection. Journal of Analytical Toxicology, 28, 357-363. http://dx.doi.org/10.1093/jat/28.5.357
[16]	Holland, K.E., Solano, M.I., Johnson, R.C., Maggio, V.L. and Barr, J.R. (2008) Modifications to the Organophosphorus Nerve Agent-Protein Adduct Refluoridation Method for Retrospective Analysis of Nerve Agent Exposures. Journal of Analytical Toxicology, 32, 116-124. http://dx.doi.org/10.1093/jat/32.1.116
[17]	Polhuijs, M., Langenberg, J.P. and Benschop, H.P. (1997) New Method for Retrospective Detection of Exposure to Or- ganophosphorus Anticholinesterases: Application to Alleged Sarin Victims of Japanese Terrorists. Toxicology and Applied Pharmacology, 146, 156-161. http://dx.doi.org/10.1006/taap.1997.8243
[18]	Adams, T.K., Capacio, B.R., Smith, J.R., Whalley, C.E. and Korte, W.D. (2004) The Application of the Fluoride Reactivation Process to the Detection of Sarin and Soman Nerve Agent Exposures in Biological Samples. Drug and Chemical Toxicology, 27, 77-91. http://dx.doi.org/10.1081/DCT-120027901
[19]	Jakubowski, E.M., Heykamp, L.S., Durst, H.D. and Thomson, S.A. (2001) Preliminary Studies in the Formation of Ethyl Methylphosphonofluoridate from Rat and Human Serum Exposed to Vx and Treated with Fluoride Ion. Analytical Letters, 34, 727-737. http://dx.doi.org/10.1081/AL-100103215
[20]	Muller, A.J. (1988) The Use of Organosilicon Esters for the Synthesis of Alkyl Phosphonofluoridates. The Journal of Organic Chemistry, 53, 3364-3365. http://dx.doi.org/10.1021/jo00249a046
[21]	Gupta, H.K., Pardasani, D., Mazumder, A., Purohit, A.K. and Dubey, D.K. (2009) Tetrabutylammonium Tetra (tert- Butyl Alcohol) Coordinated Fluoride—An Efficient Reagent for the Synthesis of Fluorine Derivatives of Phosphorus(V) Compounds. Tetrahedron Letters, 50, 2697-2699. http://dx.doi.org/10.1016/j.tetlet.2009.03.151