Correlation of Mass Spectrometry Identified Bacterial Biomarkers from a Fielded Pyrolysis-Gas Chromatography-Ion Mobility Spectrometry Biodetector with the Microbiological Gram Stain Classification Scheme

Abstract

Public reporting burden for fthi collection of Informaiaon is estimated to, average I hour per response, including the time for reviewing Instructions. searchilng erristing data sources, gatheing and maintsteng the data needed, and completing and review"n this collection of Information. Send cofmment regarding this burden estimate or any othier aspect of this collection of Infomuation, Indluding suggestions for reducing fti burden to Departmrent PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) AND ADDRESS(ES) 8. PERFORMING ORGANIZATION REPORTDIR ECBC, ATTN: AMSRD-ECBC-RT-II, APG, MD 21010-5424 NUMBER GEO-CENTERS, Inc., Gunpowder Branch, APG, MD 21010-5424 ECBC-TR-415 SPONSORING / MONITORING AGENCY NAME(S) AND ADDRESS(ES) 10. SPONSORIMONITOR'S ACRONYM(S) SPONSORIMONITOR'S REPORT NUMBER(S) DISTRIBUTION I AVAILABILITY STATEMENTApproved for public release; distribution is unlimited. SUPPLEMENTARY NOTES ABSTRACTThis work discusses the significance of the pyrolyzate peaks observed in the gas chromatography-ion mobility spectrometry (GC-IMS) dataspace of the pyrolysis-GC-IMS (Py-GC-IMS) briefcase system. This system has the ability to detect and classifyi deliberately released bioaerosols in outdoor field scenarios. The bioaerosols include Gram-positive spores and Gram-negative bacteria, the MS-2 Escherichia coliphage virus, and ovalbumnin (OV) protein species. The work suggests certain improvements that can be made to the IMS detection System. A pyrolysis-gas chromatography-ion mobility spectrometry (Py-GC-IMS) briefcase system can detect and classify deliberately released bioaerosols in outdoor field scenarios. The bioaerosols include Gram-positive spores and Gram-negative bacteria, the MS-2 Escherichia coliphage virus, and ovalbumin (OV) protein species. However, the origin and structural identities of the pyrolyzate peaks observed in the GC-IMS dataspace, their microbiological information content, and taxonomic importance with respect to biodetection have not been determined. SUBJECT TERMS PyrolysisThe present work interrogates the identities of the peaks by inserting a time-offlight (TOF) mass spectrometry (MS) system in parallel with the IMS detector through a Tee connection in the GC module. Biological substances, producing ion mobility peaks from the pyrolysis of microorganisms, have been identified by their GC retention times, by matching their electron ionization mass spectra with authentic standards, and by the National Institute of Standards and Technology (NIST) mass spectral database.Strong signals from 2-pyridinecarboxamide were identified in Bacillus samples, including Bacillus anthracis, and their origins were traced to the cell wall peptidoglycan macromolecule. 3-Hydroxymyristic acid is a component of lipopolysaccharides (LPS) in the cell walls of Gram-negative organisms. The Gram-negative E. coli organism showed significant amounts of 3-Hydroxymyristic acid derivatives and degradation products in Py-GC-MS analyses. Some of the fatty acid derivatives were observed in very low abundance in the ion mobi...

A pirólise como técnica analítica

“…Os pirogramas obtidos para os três tipos de bactérias estudados foram analisados, encontrando-se sinais característicos (biomarcardores) de tipos diferentes de bactérias, como o ácido tetradecanóico, tridecan-2-ona e dodecanal, que são compostos usados para distinguir bactéria gram-negativa (E. coli) de outras bactérias gram-positivas, como M. luteus e B. megaterium. 52 Alves et al 53 aplicaram a pirólise analítica para determinar o conteúdo de lignina em madeira de duas espécies de pinho Marítimo (Pinus pinaster e Picea abies). Os resultados obtidos com a pirólise analítica foram compatíveis com os teores de lignina Klason determinados pelo método (TAPPI T 222 OM-88), que é largamente utilizado como padrão para esse tipo de análise.…”

Feasibility Study of Using Short Wave Infrared Cavity Ringdown Spectroscopy (SWIR-CRDS) for Biological Agent Detection

“…[24][25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40] Some of the products have extremely high molecular weight, but there are also a number of low molecular weight, high volatility species that are produced. As would be expected, the lower molecular weight species prevail when the pyrolysis temperature is high and the residence time is long.…”

Quantitative assessment and optimization of parameters for pyrolysis of bacteria with gas chromatographic analysis

“…Comparisons of the chromatograms from the bacteria showed that some peaks, or constituents in the chromatograms were observed in two or more strains of bacteria and should be associated with parts of the bacterial cells found generally in bacteria, e.g. 2-furancarboxaldehyde, a product from the pyrolysis of carbohydrates or phenol which is derived from proteins [29]. These chemicals may be useful in a cursory survey of biological versus non-biological, and might serve to indicate certain bacteria.…”