Analysis of degradation products of Novichok agents in human urine by hydrophilic interaction liquid chromatography–tandem mass spectrometry
Tóm tắt
The detection of hydrolysis products of Novichok agents in biological samples from victims is important for confirming exposure to these agents. However, Novichok agents are new class of nerve agent and there have been only few reports on analyses of Novichok agent degradation products. Here, we developed hydrophilic interaction liquid chromatography (HILIC)–tandem mass spectrometry (MS/MS) methods to detect Novichok agent degradation products in human urine with simple pretreatment and high sensitivity. A Poroshell 120 HILIC-Z column was used to analyze six Novichok agent degradation products. For urine samples, we used a simple pretreatment method, which consisted of deproteinization with acetonitrile and microfiltration. We calculated the pKa values of the OH groups, the log P values, and the molecular weights to investigate the difference in chromatographic behaviors of the Novichok agent degradation products and the degradation products of conventional nerve agents. Six Novichok agent degradation products, including N-(bis-(diethylamino)methylidene)-methylphosphonamidic acid (MPGA), which could not be detected by our previous method, could be analyzed with sufficient peak shape and mutual separation. The detection limits of six Novichok agent degradation products were sufficiently low (1–50 ng/mL) and the calibration curves showed sufficient linearity. The physicochemical parameters of Novichok agent degradation products were different from those of conventional nerve agent degradation products, and this explains the difference in chromatographic behaviors. Six Novichok agent degradation products were successfully analyzed by HILIC–MS/MS. Due to the absence of a derivatization step, throughput performance was higher than our previous derivatization-liquid chromatography–MS/MS method.
Tài liệu tham khảo
Kassa J (2002) Review of oximes in the antidotal treatment of poison-ing by organophosphorus nerve agents. J Toxicol Cli Toxicol 40:803–816
Organization for the Prohibition of chemical weapons, chemical weapon convention. https://www.opcw.org. Accessed 15 June 2022
Seto Y, Tsunoda N, Kataoka M, Tsuge K, Nagano T (1999) In Natural and Selected Synthetic Toxins: Biological Implications; Tu, A. A., Gaffield, A., (Eds).; American Chemical Society: Washington, DC, pp. 318–332.
Noort D, Hulst AG, Platenburg DHJM, Polhuijs M, Benschop HP (1998) Quantitative analysis of O-isopropyl methylphosphonic acid in serum samples of Japanese citizens allegedly exposed to sarin: estimation of internal dosage. Arch Toxicol 72:671–675
Tsuchihashi H, Katagi M, Nishikawa M, Tatsuno M (1998) Identification of Metabolites of Nerve Agent VX in Serum Collected from a Victim. J Anal Toxicol 22:383–388
Summary of the Report on Activities Carried Out in Support of a Request for Technical Assistance by the United Kingdom of Great Britain and Northern Ireland (Technical Assistance Visit TAV/02/18), OPCW (2020)
Summary of the Report on Activities Carried Out In Support of a Request for Technical Assistance by Germany (Technical Assistance Visit – TAV/01/20), OPCW (2020)
Carlsen L (2019) After salisbury nerve agents revisited. Mol Inf 38:1800106
Bhakhoa H, Rhyman L, Ramasami P (2019) Theoretical study of the molecular aspect of the suspected novichok agent A234 of the Skripal poisoning. R Soc Open Sci 6:181831
Imrita Y A, Bhakhoa H, Sergeievab T, Dan´esb S, Savoo N, Elzagheid M I, Rhyman L, Andrada D M, Ramasami P ,(2020) A theoretical study of the hydrolysis mechanism of A-234; the suspected novichok agent in the Skripal attack. RSC Adv 10: 27884–27893.
Harvey SP, McMahon LR, Berg FJ (2020) Hydrolysis and enzymatic degradation of Novichok nerve agents. Heliyon 6:e03153
Mirbabaei F, Mohammad-Khah A, Naseri MT, Babri M, Faraz SM, Hosseini SE, Ashraf D (2022) Unambiguous identification and determination of A234-Novichok nerve agent biomarkers in biological fluids using GC–MS/MS and LC–MS/MS. Anal Bioanal Chem 414:3429–3442
Black RM, Muir B (2003) Derivatisation reactions in the chromatographic analysis of chemical warfare agents and their degradation products. J Chromatogr A 1000:253–281
Jang Y J, Kim K, Tsay O G, Atwood D A, Churchill D G (2015) Destruction and detection of chemical warfare agents. Chem Rev 115: PR1–PR76.
Noort D (2021) Fidder A van der Riet-van O D, Busker R, van der Schans M J (2021) verification of exposure to Novichok nerve Agents Utilizing a Semitargeted Human Butyrylcholinesterase Nonapeptide assay. Chem Res in Toxicol 34:1926–1932
Lee JY, Lim KC, Kim HS (2021) Characterization and study on fragmentation pathways of a novel nerve agent, ‘Novichok (A234)’, in aqueous solution by liquid chromatography-tandem mass spectrometry. Molecules 26:1059
Otsuka M, Miyaguchi H (2021) Theoretical evaluation of the hydrolysis of conventional nerve agents and novichok agents. Chem Phys Lett 785:139116
Yamaguchi A, Miyaguchi H, Tokeshi M (2022) Dimethoxytriadinylation LC-MS/MS of novichok a-series degradation products in human urine. Anal Chem 94:4658–4665
Dubey V, Velikeloth S, Sliwakowski M, Mallard G (2009) Official proficiency tests of the organisation for the prohibition of chemical weapons: current status and future directions. Accred Qual Assur 14:431–437
Quality System Document of the OPCW: Work Instruction for the Reporting of the Results of the OPCW Biomedical Proficiency tests (QDOC_LAB_WI_BioPT04 (Issue 1, Revision 1, dated 28 December 2016)).
Tetko IV, Gasteiger J, Todeschini R, Mauri A, Livingstone D, Ertl P, Palyulin VA, Radchenko EV, Zefirov NS, Makarenko AS, Tanchuk VY, Prokopenko VV (2005) Virtual computational chemistry laboratory-design and description. J Comput Aid Mol Des 19:453–463
VCCLAB, Virtual Computational Chemistry Laboratory, http://www.vcclab.org, 2005 (Accessed 8 August 2022).
Gaussian 09, Revision C.01 or D.01, Frisch, M. J. et al. Gaussian, Inc., Wallingford CT, 2009.
Gaussian 16, Revision C.01, Frisch, M. J. et al. Gaussian, Inc., Wallingford CT, 2016.
Yang C, Xue XS, Jin JL, Li X, Cheng JP (2013) Theoretical Study on the Acidities of Chiral Phosphoric Acids in Dimethyl Sulfoxide: Hints for Organocatalysis. J Org Chem 78:7076–7085
Becke AD (1988) Density-functional exchange-energy approximation with correct asymptonic behavior. Phys Rev A 38:3098–3100
Becke AD (1993) A new mixing of Hartree-Fock and local-density-functional theories. J Chem Phys 98:1372–1377
Becke AD (1993) Density-functional thermochemistry. III. the role of exact exchange. J Chem Phys 98:5648–5652
Lee C, Yang W, Parr RG (1988) Development of the colle-Salvetti correlation-energy formula into a functional of the electron density. Phys Rev B 37:785–789
Zhao Y, Truhlar DG (2008) The M06 suite of density functionals for main group thermochemistry, thermochemical kinetics, noncovalent interactions, excited states, and transition elements: two new functionals and systematic testing of four M06-class functionals and 12 other functionals. Theor Chem Account 120:215–241
http://ccc.chem.pitt.edu/wipf/MechOMs/evans_pKa_table.pdf. Accessed 8 Aug 2022
Otsuka M, Miyaguchi H (2022) Analysis of degradation products of nerve agents in biological fluids by ion chromatography–tandem mass spectrometry. Forensic Toxicol In press. https://doi.org/10.1007/s11419-022-00633-x)
Palumbo D, Fais P, Calì A, Lusardì M, Bertol E, Pascalì JP (2018) Novel zwitterionic HILIC stationary phase for the determination of ethyl glucuronide in human hair by LC-MS/MS. J Chromatogr B 1100–1101:33–38
Otsuka M, Miyaguchi H, Uchiyama M (2019) Analysis of degradation products of nitrogen mustards via hydrophilic interaction liquid chromatography–tandem mass spectrometry. J Chromatogr A 1602:199–205
HILIC Trouble shooting, Thermo Fisher Scientific, https://www.thermofisher.com/jp/en/home/industrial/chromatography/chromatography-learning-center/liquid-chromatography-information/hilic-hplc-uhplc-columns-information/hilic-troubleshooting.html (Accessed 6 September).
Otsuka M, Tsuge K, Seto Y, Miyaguchi H, Uchiyama M (2018) Analysis of degradation products of nerve agents via post-pentafluorobenzylation liquid chromatography-tandem mass spectrometry. J Chromatogr A 1577:31–37