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Rationale  Rapid sub‐species characterization of pathogens is required for timely responses in outbreak situations. Pyrolysis mass spectrometry (PyMS) has the potential to be used for this purpose.    Methods  However, in order to make PyMS practical for traceback applications, certain improvements related to spectrum reproducibility and data acquisition speed were required. The main objectives of this study were to facilitate fast detection (<30 min to analyze 6 samples, including preparation) and sub‐species‐level bacterial characterization based on pattern recognition of mass spectral fingerprints acquired from whole cells volatilized and ionized at atmospheric pressure. An AccuTOF DART mass spectrometer was re‐engineered to permit ionization of low‐volatility bacteria by means of  Plasma Jet Ionization  (PJI), in which an electric discharge, and, by extension, a plasma beam, impinges on sample cells.    Results  Instrumental improvements and spectral acquisition methodology are described. Performance of the re‐engineered system was assessed using a small challenge set comprised of assorted bacterial isolates differing in identity by varying amounts. In general, the spectral patterns obtained allowed differentiation of all samples tested, including those of the same genus and species but different serotypes.    Conclusions  Fluctuations of ±15% in bacterial cell concentrations did not substantially compromise replicate spectra reproducibility. © 2015 National Center for Toxicological Research.  Rapid Communications in Mass Spectrometry  published by John Wiley & Sons Ltd.

Instrumental improvements and sample preparations that enable reproducible, reliable acquisition of mass spectra from whole bacterial cells