Monitoring Exposure to Five Chemical Warfare Agents Using the Dried Urine Spot Technique and Liquid Chromatography-Mass Spectrometry/Mass Spectrometry—In Vivo Determination of Sarin Metabolite in Mice
Abstract
:1. Introduction
2. Results
2.1. Development and Optimization of the Extraction Procedure (Urine Volume, “In-Tip” Extraction Method, Solvent and Volume)
2.2. Validation of the Method
2.2.1. Calibration Model
2.2.2. Selectivity (Interferences)
2.2.3. LOD and LOQ in Urine Samples
2.2.4. Recoveries
2.2.5. Accuracy and Precision
2.2.6. Stability
2.2.7. Matrix Effects in LC-MS/MS
2.2.8. DUS vs. “Dilute and Shoot”
2.3. Exposure of Mice to GB—In Vivo Experiment
2.4. Non-Targeted “Screening”
3. Summary and Conclusions
4. Materials and Methods
4.1. Chemicals and Reagents
4.2. Preparation of Standard Solutions
4.3. Instrument Conditions
4.4. DUS Sample Preparation
4.5. Animals
4.6. In Vivo Experiment
4.7. Method Validation
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Conflicts of Interest
References
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Compound | Dominant MRM Transition (Negative ESI) | tR (min) | LOD (ng/mL) | LOQ (ng/mL) |
---|---|---|---|---|
EMPA (VX acid) | 123 > 95 | 2.7 | 10 | 30 |
IMPA (GC-acid) | 137 > 79 | 3.7 | 3 | 9 |
iBuMPA (RVX-acid) | 151 > 77 | 5.5 | 1 | 3 |
CMPA (GF-acid) | 177 > 95 | 6.6 | 0.5 | 3 |
PMPA (GD-acid) | 179 > 95 | 7.8 | 0.5 | 3 |
Compound | 10 ng/mL in Urine | 100 ng/mL in Urine | ||||
---|---|---|---|---|---|---|
Recovery (%) | Accuracy (%) | Precision (%) | Recovery (%) | Accuracy (%) | Precision (%) | |
EMPA | 41.7 | 23.1 | 9.2 | 40 | 21.2 | 12.5 |
IMPA | 53.3 | 20.6 | 8.5 | 44 | 19.7 | 12.5 |
iBuMPA | 55.5 | 14.6 | 8.9 | 80 | 12.6 | 11.7 |
CMPA | 49.1 | 24.4 | 8.1 | 56 | 19.1 | 13.3 |
PMPA | 54.1 | 16.3 | 8.5 | 54 | 1.9 | 2.9 |
Characteristics | DBS [19] | DUS (This Study) |
---|---|---|
Sample volume, µL | 20 | 50 |
Sample preparation | Extract: 400 µL MeOH Evaporation to dryness, add 400 µL water | Extract (in a polypropylene tip): 300 µL MeOH:H2O Filter the extract |
LOD (ng/mL): | ||
EMPA (VX acid) | 1 | 10 |
IMPA (GB—acid) | 0.5 | 3 |
iBuMPA (RVX-acid) | 1 | 1 |
CMPA (GF-acid) | 1 | 0.5 |
PMPA (GD-acid) | 0.3 | 0.5 |
Long-term sample stability on the paper | ˃~1 month | ˃5 months |
Concentration in an in vivo experiment (ng/mL): | 1LD50, 5 rats, IM.: | 1LD50, 5 mice, IN.: |
2 h after the exposure (Avg) | 28 | 8800 |
20 h after the exposure (Avg) | 3700 | |
24 h after the exposure (Avg) | 3.3 |
Metabolite Structure | Negative Ion Formula | MRM (ESI−) | Transition Intensity Ratio |
---|---|---|---|
EMPA (VX Acid) | C3H9PO3− | 123.0 > 95.0 | 3 |
123.0 > 79.0 | 1 | ||
IMPA (GB Acid) | C4H11PO3− | 137.0 > 95.0 | 6 |
137.0 > 79.0 | 1 | ||
iBuMPA (RVX Acid) | C5H13PO3− | 151.1 > 77.0 | 2 |
151.1 > 79.0 | 1 | ||
CMPA (GF Acid) | C7H15PO3− | 177.1 > 95.0 | 2 |
177.1 > 79.0 | 1 | ||
PMPA (GD Acid) | C7H17PO3− | 179.1 > 95.0 | 2 |
179.1 > 79.0 | 1 |
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Yishai Aviram, L.; Dagan, S.; Hindi, A.; Chapman, S.; Gez, R.; Drug, E. Monitoring Exposure to Five Chemical Warfare Agents Using the Dried Urine Spot Technique and Liquid Chromatography-Mass Spectrometry/Mass Spectrometry—In Vivo Determination of Sarin Metabolite in Mice. Molecules 2023, 28, 7687. https://doi.org/10.3390/molecules28237687
Yishai Aviram L, Dagan S, Hindi A, Chapman S, Gez R, Drug E. Monitoring Exposure to Five Chemical Warfare Agents Using the Dried Urine Spot Technique and Liquid Chromatography-Mass Spectrometry/Mass Spectrometry—In Vivo Determination of Sarin Metabolite in Mice. Molecules. 2023; 28(23):7687. https://doi.org/10.3390/molecules28237687
Chicago/Turabian StyleYishai Aviram, Lilach, Shai Dagan, Ariel Hindi, Shira Chapman, Rellie Gez, and Eyal Drug. 2023. "Monitoring Exposure to Five Chemical Warfare Agents Using the Dried Urine Spot Technique and Liquid Chromatography-Mass Spectrometry/Mass Spectrometry—In Vivo Determination of Sarin Metabolite in Mice" Molecules 28, no. 23: 7687. https://doi.org/10.3390/molecules28237687
APA StyleYishai Aviram, L., Dagan, S., Hindi, A., Chapman, S., Gez, R., & Drug, E. (2023). Monitoring Exposure to Five Chemical Warfare Agents Using the Dried Urine Spot Technique and Liquid Chromatography-Mass Spectrometry/Mass Spectrometry—In Vivo Determination of Sarin Metabolite in Mice. Molecules, 28(23), 7687. https://doi.org/10.3390/molecules28237687