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Raman spectroscopy, electronic microscopy and SPME-GC-MS to elucidate the mode of action of a new antimicrobial food packaging material

Analytical and Bioanalytical Chemistry volume 409, pages 1037–1048 (2017)Cite this article

Abstract

One critical challenge when developing a new antimicrobial packaging material is to demonstrate the mode of action of the antimicrobials incorporated into the packaging. For this task, several analytical techniques as well as microbiology are required. In this work, the antimicrobial properties of benzyl isothiocyanate, allyl isothiocyanate and essential oils of cinnamon and oregano against several moulds and bacteria have been evaluated. Benzyl isothiocyanate showed the highest antimicrobial activity and it was selected for developing the new active packaging material. Scanning electron microscopy and Raman spectroscopy were successfully used to demonstrate the mode of action of benzyl isothiocyanate on Escherichia coli. Bacteria exhibited external modifications such as oval shape and the presence of septum surface, but they did not show any disruption or membrane damage. To provide data on the in vitro action of benzyl isothiocyanate and the presence of inhibition halos, the transfer mechanism to the cells was assessed using solid-phase microextraction–gas chromatography–mass spectrometry. Based on the transfer system, action mechanism and its stronger antimicrobial activity, benzyl isothiocyanate was incorporated to two kinds of antimicrobial labels. The labels were stable and active for 140 days against two mould producers of ochratoxin A; Penicillium verrucosum is more sensitive than Aspergillus ochraceus. Details about the analytical techniques and the results obtained are shown and discussed.

Antimicrobial evaluation of pure compounds, incorporation in the packaging and study for mode of action on S. coli by Raman, SEM and SPME-GC-MS

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Acknowledgments

This work was supported by University of Zaragoza (PIFUZ-2012-B-CIE-001) within the scope of 2012/0254 REPSOL lubricantes y especialidades (Rylesa). Thanks are also given to Gobierno de Aragón and Fondo Social Europeo for the financial help of GUIA Group, T-10. Warm and sincere thanks are given to Thermo Fisher Scientific for the successful and efficient collaboration. Margarita Aznar acknowledges the Spanish Ministry of Economy for its Ramon y Cajal contract (Project RYC-2012-11856). The authors acknowledge Carlos Cuestas from the Advanced Microscopy Laboratory (LMA) of the Aragón Institute of Nanoscience (INA) for the technical support provided during SEM analysis.

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  1. Aragón Institute of Engineering Research (I3A), EINA, Department of Analytical Chemistry, University of Zaragoza, María de Luna 3 Torres Quevedo Bldg., 50018, Zaragoza, Spain

    Isabel Clemente, Margarita Aznar, Jesús Salafranca & Cristina Nerín

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  1. Isabel Clemente
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Correspondence to Cristina Nerín.

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Clemente, I., Aznar, M., Salafranca, J. et al. Raman spectroscopy, electronic microscopy and SPME-GC-MS to elucidate the mode of action of a new antimicrobial food packaging material. Anal Bioanal Chem 409, 1037–1048 (2017). https://doi.org/10.1007/s00216-016-0022-y

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Keywords

  • Essential oil
  • Benzyl isothiocyanate
  • Raman spectroscopy
  • Scanning electron microscopy
  • SPME-GC-MS
  • Antimicrobial active packaging