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Rapid detection and quantitation of dipicolinic acid from Clostridium botulinum spores using mixed-mode liquid chromatography-tandem mass spectrometry

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Abstract

Analysis of the dipicolinic acid (DPA) released from Clostridium botulinum spores during thermal processing is crucial to obtaining a mechanistic understanding of the factors involved in spore heat resistance and related food safety applications. Here, we developed a novel mixed-mode liquid chromatography-tandem mass spectrometry (LC–MS/MS) method for detection of the DPA released from C. botulinum type A, nonproteolytic types B and F strains, and nonpathogenic surrogate Clostridium sporogenes PA3679 spores. DPA was retained on a mixed-mode C18/anion exchange column and was detected using electrospray ionization (ESI) positive mode within a 4-min analysis time. The intraday and interday precision (%CV) was 1.94–3.46% and 4.04–8.28%, respectively. Matrix effects were minimal across proteolytic type A Giorgio-A, nonproteolytic types QC-B and 202-F, and C. sporogenes PA3679 spore suspensions (90.1–114% of spiked DPA concentrations). DPA recovery in carrot juice and beef broth ranged from 105 to 118%, indicating limited matrix effects of these food products. Experiments that assessed the DPA released from Giorgio-A spores over the course of a 5-min thermal treatment at 108 °C found a significant correlation (R = 0.907; P < 0.05) between the log reduction of spores and amount of DPA released. This mixed-mode LC–MS/MS method provides a means for rapid detection of DPA released from C. botulinum spores during thermal processing and has the potential to be used for experiments in the field of food safety that assess the thermal resistance characteristics of various C. botulinum spore types.

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Abbreviations

DPA:

Dipicolinic acid

ESI:

Electrospray ionization

LC-MS:

Liquid chromatography-mass spectrometry

MS:

Mass spectrometry

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Funding

The research described in this publication is supported by the Food and Drug Administration (FDA) of the U.S. Department of Health and Human Services (HHS) as part of an award totaling $4,148,332 with 0% financed with non-governmental sources. The contents are those of the authors and do not necessarily represent the official views of, nor an endorsement, by FDA, HHS, or the U.S. Government. For more information, please visit FDA.gov.

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Authors and Affiliations

  1. Division of Food Processing Science and Technology, Office of Food Safety, U.S. Food and Drug Administration, 6502 South Archer Road, Bedford Park, IL, USA

    Benjamin W. Redan, Travis R. Morrissey, Catherine A. Rolfe & N. Rukma Reddy

  2. Institute for Food Safety and Health, Illinois Institute of Technology, 6502 South Archer Road, Bedford Park, IL, USA

    Viviana L. Aguilar

  3. Multi-Component Foods, Food Process Evaluation Team, Office of Food Safety, U.S. Food and Drug Administration, 5001 Campus Drive, College Park, MD, USA

    Guy E. Skinner

Authors
  1. Benjamin W. Redan
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  2. Travis R. Morrissey
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  3. Catherine A. Rolfe
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  4. Viviana L. Aguilar
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  5. Guy E. Skinner
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  6. N. Rukma Reddy
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Correspondence to Benjamin W. Redan.

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Redan, B.W., Morrissey, T.R., Rolfe, C.A. et al. Rapid detection and quantitation of dipicolinic acid from Clostridium botulinum spores using mixed-mode liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 414, 2767–2774 (2022). https://doi.org/10.1007/s00216-022-03926-7

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  • DOI: https://doi.org/10.1007/s00216-022-03926-7

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