Chemical Warfare Agents by LC-MS/MS. Don Noot _ Vogon Labs Michele Mayer _ DRDC Suffield VLS. Vogon Laboratory Services. Analytical Consulting
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1 Chemical Warfare Agents by LC-MS/MS Don Noot _ Vogon Labs Michele Mayer _ DRDC Suffield
2 Working on the base in Suffield Tried to get a picture on the base... 2
3 Venn Diagram #1 3
4 Venn Diagram #2 4
5 Venn Diagram #2 Vancouver version 5
6 CWA Venn Diagram 6
7 CWA Method of Analysis many methods for CWA historical methods using GC-MS, derivatization required for breakdown products more recently, LC-MS methods allowing detection without derivatization 7
8 CWA Method of Analysis - 2 existing methods for environmental samples (water, soil, extracts of surfaces) biological fluids needed a method for CWAs in decontamination formulations 8
9 Decontamination Formulations the problem real time analysis of agent in complex matrix the matrix solvent of some type to dissolve agent; reactive ingredient; possible counter ion (e.g. Na) 9
10 Decontamination Formulations - 2 need to deactivate the active ingredient quickly to capture a agent concentration in solution stop the reaction direct analysis by dilute and shoot LC-MS/MS should be perfect 10
11 G Agents - GF Cyclohexyl sarin Cyclohexyl methylphosphonofluoridate 11
12 G Agents - GD Soman o-pinacolyl methylphosphonofluoridate 12
13 ISTD tough to get isotopically labeled CW agents! TEP, Triethyl phosphate resistant to hydrolysis 13
14 MS/MS Optimization 3rd step in working with new compound is optimization of MS/MS parameters 2nd step: run full scan do I have what I think I have in the vial? also run a blank! 1 st step? 14
15 TEP in ACN - FIA 15
16 MS/MS Optimization - GF GF by FIA didn t look like I had GF in the vial very small [M+H] + lots of other ions 16
17 GF in ACN flow injection analysis 17
18 FIA negative peaks? 18
19 GF in ACN fast chromatography 19
20 GF in decon formulation 20
21 GF - Effect of NH 4 Ac in ESI 21
22 GF & TEP Effect of MeOH vs ACN 22
23 GF & TEP in MPEG matrix 23
24 TEP good indicator of ion suppression 24
25 APCI vs ESI ESI shows greater sensitivity (AJS) APCI less susceptible to chromatographic additives and matrix Injected [M+NH4]+ to [M+H]+ Area Ratio ESI APCI GF ng 15.7 GF ng GF 0.12 ng GF 0.6 ng GF 3 ng average Relative intensity of [M+NH4]+ to [M+H]+ for GF by ESI and APCI 25
26 GF - Effect of NH 4 Ac in APCI 26
27 GF - Matrix Effects in APCI 27
28 ESI vs APCI final method chosen was ESI why, if APCI shows less variability from matrix and effects chromatographic conditions? ESI-AJS was more precise dilute matrix out to level where no ion suppression is also good for decon reaction quenching use TEP as indicator of ion suppression increased confidence in results 28
29 ESI vs APCI - precision Name Acq. Date-Time TEP Area GF-NH4 Area GF & TEP Std 2010/1/ GF & TEP Std 2010/1/ GF & TEP Std 2010/1/ GF & TEP Std 2010/1/ GF & TEP Std 2010/1/ GF & TEP Std 2010/1/ GF & TEP Std 2010/1/ GF & TEP Std 2010/1/ average std dev %RSD 0.4% 1.0% Name Acq. Date-Time TEP Area GF-NH4 Area TEP & GF in AcCN 2010/3/ TEP & GF in AcCN 2010/3/ TEP & GF in AcCN 2010/3/ TEP & GF in AcCN 2010/3/ TEP & GF in AcCN 2010/3/ TEP & GF in AcCN 2010/3/ TEP & GF in AcCN 2010/3/ TEP & GF in AcCN 2010/3/ TEP & GF in AcCN 2010/3/ TEP & GF in AcCN 2010/3/ average std dev %RSD 4.7% 5.5% 29
30 GF linearity using ESI-AJS 30
31 GF - Instrument Detection Limit approximately 8 pg on-column 31
32 Decon Formulation Experiment Sample TEP Results GF-NH4 Results TPP Results Name RT Resp. RT Resp. RT Resp. GF TEP TPP in ACN RSDL decon_gf_93mm11-1a RSDL decon_gf_93mm11-1b RSDL decon_gf_93mm11-1c RSDL decon_gf_93mm11-1d GF TEP TPP in ACN GF TEP TPP in ACN RSDL decon_gf_93mm11-1a RSDL decon_gf_93mm11-1b RSDL decon_gf_93mm11-1c RSDL decon_gf_93mm11-1d GF TEP TPP in ACN n average std dev %RSD 0.07% 3.54% 0.09% 1.66% 0.08% 4.14% 32
33 Conclusions dilute and shoot LC-MS/MS method is appropriate for CWA studies in decontamination formulations quench decon reaction dilute out matrix use ESTD calculation TEP works well as indicator of ion suppression TPP works well as surrogate 33
34 34
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