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4. Combining presumptive color tests, pressure-sensitive adhesive-based collection, and paper spray-mass spectrometry for illicit drug detection.

Author

Prunty, Sarah, Carmany, Daniel, Dhummakupt, Elizabeth S., and Manicke, Nicholas E.

Subjects
*DRUGS of abuse, *HEROIN, *METHAMPHETAMINE, *DRUG residues, *PRESSURE-sensitive adhesives, *DRUG traffic, *SPECTROMETRY, *ELECTROSPRAY ionization mass spectrometry
Abstract

Illicit drug trafficking and abuse is a significant public safety and health concern. Color tests are commonly used for drug screening, but their poor specificity results in false positives. This study demonstrates the combination of drug residue collection using pressure-sensitive adhesive paper, on-paper color testing, and post-reaction analysis by paper spray mass spectrometry (PS-MS) on both portable and benchtop ion trap MS. All steps, including residue collection, color testing, and paper spray analysis, were performed on the same piece of paper. Three common color tests were investigated: the cobalt thiocyanate test for cocaine, the Simon test for methamphetamine, and the Marquis test for phenethylamine stimulants and opiates. The detection threshold for color tests ranged from 1.25 to 10 μg on paper. Drug residues were successfully confirmed by paper spray MS at the color test threshold in all cases, except for heroin after reaction with the Marquis reagent, when using the portable MS. In this case, the MS detection threshold was 4-fold higher than the color test threshold. The stability of the color test products was assessed through a time study. Drug residues could be detected by MS at least 24 hours after reaction. A series of realistic samples, including false positives, were analyzed to demonstrate the technique's utility in real-world scenarios. Overall, combining color tests with PS-MS offers a rapid, low-cost method for the collection and analysis of illicit drugs. [ABSTRACT FROM AUTHOR]

Published
2023
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10. Paper spray ionization: Applications and perspectives

Author

McBride, Ethan M., primary, Mach, Phillip M., additional, Dhummakupt, Elizabeth S., additional, Dowling, Sarah, additional, Carmany, Daniel O., additional, Demond, Paul S., additional, Rizzo, Gabrielle, additional, Manicke, Nicholas E., additional, and Glaros, Trevor, additional

Published
2019
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12. Cover Image

Author

Mach, Phillip M., primary, Dhummakupt, Elizabeth S., additional, Carmany, Daniel O., additional, McBride, Ethan M., additional, Busch, Michael W., additional, Demond, Paul S., additional, Rizzo, Gabrielle M., additional, Hollinshead, Donna E., additional, and Glaros, Trevor, additional

Published
2018
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14. Direct Analysis of Aerosolized Chemical Warfare Simulants Captured on a Modified Glass-Based Substrate by ‘Paper-Spray’ Ionization

Author

Dhummakupt, Elizabeth, Mach, Phillip M, Carmany, Daniel, Demond, Paul, Moran, Theodore, Connell, Theresa, Wylie, Harold, Manicke, Nicholas E, Nilles, J. Michael, and Glaros, Trevor

Abstract

Paper spray ionization mass spectrometry (PS-MS) offers a rapid alternative platform requiring no sample preparation. Aerosolized CWA simulants, trimethyl phosphate, dimethyl methylphosphonate, and diisopropyl methylphosphonate, were captured by passing air through a glass fiber filter disk and simultaneously capturing it directly onto a disposable paper spray cartridge. CWA simulants were aerosolized at varying concentrations using an in-house built aerosol chamber. A custom 3D-printed holder was designed and built to facilitate the aerosol capture onto the paper spray cartridges. The air flow through each of the collection devices was maintained equally to assure the same volume of air sampled across methods. Each approach yielded linear calibration curves with R2 values between 0.98–0.99 for each compound and similar limits of detection in terms of disbursed aerosol concentration. While the glass fiber filter disk has a higher capture efficiency (≈40%), the paper spray method produces analogous results even with a lower capture efficiency (≈1%). Improvements were made to include glass fiber filters as the substrate within the paper spray cartridge consumable. Glass fiber filters were then treated with ammonium sulfate to decrease chemical interaction with the simulants. This allowed for improved direct aerosol capture efficiency (>40%). Ultimately, the limits of detection were reduced to levels comparable to current worker population limits of 1x10-6 mg/m3.

Published
2024
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16. Direct Analysis of Aerosolized Chemical Warfare Simulants Captured on a Modified Glass-Based Substrate by “Paper-Spray” Ionization

Author

Dhummakupt, Elizabeth S., primary, Mach, Phillip M., additional, Carmany, Daniel, additional, Demond, Paul S., additional, Moran, Theodore S., additional, Connell, Theresa, additional, Wylie, Harold S., additional, Manicke, Nicholas E., additional, Nilles, J. Michael, additional, and Glaros, Trevor, additional

Published
2017
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17. Genetic and biochemical studies of phosphatase activity of PhoR

Author

Carmany, Daniel O., Hollingsworth, Kristine, and McCleary, William R.

Subjects
Bacteriology -- Research, Phosphatases -- Physiological aspects, Genetic research -- Analysis, Biochemistry -- Research, Histidine -- Physiological aspects, Biological sciences
Abstract

Research has been conducted on Escherichia coli PhoR histidine kinase of the phosphate regulon. The hypothesis that PhoR can function as phospho-PhoB phosphatase has been tested, and the results are described.

Published
2003

18. Proteomic and Metabolomic Profiling Identify Plasma Biomarkers for Exposure to Ultra-low Levels of Carfentanil.

Author

Dhummakupt, Elizabeth S, Rizzo, Gabrielle M, Feasel, Michael, Mach, Phillip M, Tran, Bao Q, Carmany, Daniel O, Demond, Paul S, McBride, Ethan M, Maughan, Michele, Sekowski, Jennifer W, and Glaros, Trevor

Subjects
OPIOID analgesics, PROTEOMICS, METABOLOMICS, BIOLOGICAL tags, BLOOD plasma, LIQUID chromatography-mass spectrometry
Abstract

Despite the recent epidemic of fentanyl abuse, there are few validated assays capable of rapidly detecting these compounds. In order to improve the ability to detect carfentanil at physiologically relevant concentrations, we developed a systems biology approach to discover host-based markers which are specifically amplified upon exposure in a rabbit model. For this work, two "omics" pipelines utilizing mass spectrometry were developed and leveraged. First, a proteomics pipeline was developed to interrogate the blood plasma for protein-based biomarkers. Due to the incredible dynamic range of the plasma protein content, a multi-dimensional fractionation technique was used to partition and more accurately investigate the circulating plasma proteome. Isobaric tandem mass tags were integrated into the workflow to make quantitative assessments across all animals for an extended time course post-exposure. In addition to the proteomics efforts, blood plasma was also processed through an untargeted metabolomics pipeline. This approach allows for the identification of >800 small molecule features. By processing and analyzing data sets in parallel, we were able to identify a unique fingerprint of protein and metabolite perturbations that manifest following exposure to carfentanil. [ABSTRACT FROM AUTHOR]

Published
2019
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20. In Silico Docking of Small-Molecule Inhibitors to the Escherichia coli Type III Secretion System EscN ATPase

Author

ARMY EDGEWOOD CHEMICAL BIOLOGICAL CENTER APG MD, Henderson, Terry J, Carmany, Daniel O, ARMY EDGEWOOD CHEMICAL BIOLOGICAL CENTER APG MD, Henderson, Terry J, and Carmany, Daniel O

Abstract

The type III secretion system (T3SS) is a supermolecular construct that allows many Gram-negative pathogens to translocate effector virulence proteins directly into host cells in a process requiring adenosine triphosphate (ATP) hydrolysis. We used in silico molecular modeling and computational chemistry to determine the conformations of three different small-molecule compounds when bound to EscN, the T3SS ATPase of enteropathogenic Escherichia coli. Each compound was shown to separately bind EscN within or very close to its active site and to effectively inhibit catalytic activity. Two structurally related compounds were observed to adopt extended conformations in the active-site cleft and essentially occupied the entire cleft, which obstructed the ATP binding including the formation of ATP-EscN non-covalent interactions and hydrogen bonds. Both compounds appeared to span the region in the active site with Phe355 and Gln426, residues that are essential for forming -stacking interactions and hydrogen bonds with the adenosine base of the ATP substrate. In contrast, the third compound appeared to fold upon itself in the EscN active-site cleft to adopt a very compact conformation that occupied only one side of the cleft. Our goal was to determine the three-dimensional structures of the compound EscN complexes for designing lead compounds and ultimately develop broad-spectrum therapeutics against T3SS pathogens., The original document contains color images.

Published
2014

21. Purification and Characteriztion of the Type III Secretion System Protein from Burkholderia mallei

Author

ARMY EDGEWOOD CHEMICAL BIOLOGICAL CENTER APG MD, Guelta, Mark A, Dorsey, Russell, Carmany, Daniel, Swietnicki, Wieslaw, ARMY EDGEWOOD CHEMICAL BIOLOGICAL CENTER APG MD, Guelta, Mark A, Dorsey, Russell, Carmany, Daniel, and Swietnicki, Wieslaw

Abstract

Burkholderia mallei is a Gram-negative zoonotic pathogen of solipeds that can also secondarily infect human hosts causing glanders. The bacterium is highly infectious and has been considered in the past as a potential biological weapon. In Western Europe and North America, glanders has been eradicated, but it persists in other areas. The two major bacterial virulence systems, the type III (T3SS) and type VI (T6SS) secretion systems, have been studied and found to be essential for the virulence in animal models of the disease. B. mallei has two T3SSs that are plant-like and animal-like, but only the latter is essential for virulence in animal models of glanders. In this study, we have expressed, purified, and measured the catalytic activity of the B. mallei BsaS ATPase in an in vitro assay., The original document contains color images.

Published
2013

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