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Screening for Antimicrobials in Mouthwashes Using HPTLC-Bioluminescence Detection

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Abstract

One of the tasks of food law enforcement authorities is to supervise the composition of cosmetics. In the case of mouthwashes, they are likely to contain (labeled or unlabeled) antimicrobial compounds. Conventional analyses, such as high-performance liquid chromatography (HPLC) and gas chromatography (GC) only shed light on a compound’s structure, but not on its biological function. In this study, we demonstrate that the task of detecting antimicrobials in mouthwashes can be streamlined using the luminescent bacterium Vibrio fischeri as a biodetector coupled with high-performance thin-layer chromatography (HPTLC) as a pre-separation method. The employment of subsequent conventional techniques could then be restricted to fractions with proven V. fischeri toxicity. Samples were separated in parallel on silica gel and amino layer HPTLC plates, developed with a solvent system containing tertiary butyl methyl ether and n-hexane and dried on a plate heater. After applying V. fischeri onto the HPTLC plate, zones of interest were extracted from a parallel plate and identified by HPLC–UV or GC-mass spectrometry. The reaction of V. fischeri to more than 40 standard substances which might be present in mouthwashes was determined. Based on this information, six commercially available mouthwashes were analyzed. The workflow proved to be viable for an effect-directed screening for antimicrobial compounds. The analysis of mouthwashes revealed that not only declared preservatives are used (sodium benzoate, cetylpyridinium chloride) but also other compounds, especially constituents of essential oils. Because their main purpose is flavoring of the mouthwash, they are summarized as “aroma” (anethole, carvone, menthol, thymol) which is in compliance with legal restrictions.

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Acknowledgments

We thank our colleagues at the State Laboratory Basel-City, Dr. Urs Hauri and his team for the advice and the help with HPLC–DAD, and Dr. Markus Niederer and his team for the assistance with the GC–MS. Furthermore, we thank CAMAG, especially Mr. Leuenberger, for the support concerning the HPTLC equipment. We would also like to thank Mr. Handloser (CAMAG) and Mr. Schulz (Merck Millipore/Lab Solutions, R&D Thin-layer chromatography) for the information on the properties of HPTLC layers.

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Authors and Affiliations

  1. Kantonales Laboratorium Basel-Stadt (State Laboratory Basel-City), Kannenfeldstrasse 2, 4056, Basel, Switzerland

    Vera Baumgartner & Christopher Hohl

  2. Institute of Food Chemistry, University of Hohenheim, Stuttgart, Germany

    Wolfgang Schwack

Authors
  1. Vera Baumgartner
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  2. Christopher Hohl
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  3. Wolfgang Schwack
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Correspondence to Vera Baumgartner.

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Published in the topical collection Miniaturized and New Featured Planar Chromatography and Related Techniques with guest editor Paweł K. Zarzycki.

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Baumgartner, V., Hohl, C. & Schwack, W. Screening for Antimicrobials in Mouthwashes Using HPTLC-Bioluminescence Detection. Chromatographia 76, 1315–1325 (2013). https://doi.org/10.1007/s10337-012-2375-5

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  • DOI: https://doi.org/10.1007/s10337-012-2375-5

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