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An efficient approach for the isolation, identification and evaluation of antimicrobial plant components on an analytical scale, demonstrated by the example of Radix imperatoriae

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Abstract

Using Radix imperatoriae (the root of masterwort) as an example, we describe an efficient approach for the isolation, identification and evaluation of bioactive plant components on an analytical scale. The extraction of Radix imperatoriae with ethyl acetate was enhanced by the application of ultrasound oscillations. This rhizome extract was applied to three pathogenic bacteria (Bacillus cereus, Escherichia coli, and Staphylococcus aureus) to determine its antimicrobial activity. Disk diffusion was utilized to determine susceptibility. The extract components were separated using a series of chromatography approaches (semi-preparative RP-HPLC, or RP-HPLC on an analytical scale), followed by testing. All fractions were analyzed by LC-UV-ESI-MS and 600 MHz microcoil 1H NMR spectroscopy. Among other findings, in the fraction with the highest antibacterial activity we were able to identify oxypeucedanin and oxypeucedanin hydrate. Subsequent analysis revealed that only oxypeucedanin hydrate had antibacterial activity, whereas oxypeucedanin itself was inactive at the concentrations applied. Furthermore, oxypeucedanin hydrate appears to be largely, or exclusively, a by-product of sample preparation, since it is either not synthesized by the plant as a second metabolite or is produced by it in only very small quantities.

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Acknowledgements

The authors thank the Institute of Botany at the University of Tübingen for supplying them with plant material.

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Authors and Affiliations

  1. Institute of Organic Chemistry, University of Tübingen, Auf der Morgenstelle 18, 72076, Tübingen, Germany

    O. Gökay & K. Albert

  2. Institute of Microbiology and Infectious Diseases, University of Tübingen, Auf der Morgenstelle 28, 72076, Tübingen, Germany

    D. Kühner, F. Götz & U. Bertsche

  3. Interfaculty Institute for Biochemistry, University of Tübingen, Hoppe-Seyler-Strasse 4, 72076, Tübingen, Germany

    M. Los

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  1. O. Gökay
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  2. D. Kühner
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  3. M. Los
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  4. F. Götz
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  5. U. Bertsche
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  6. K. Albert
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Correspondence to K. Albert.

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Gökay, O., Kühner, D., Los, M. et al. An efficient approach for the isolation, identification and evaluation of antimicrobial plant components on an analytical scale, demonstrated by the example of Radix imperatoriae . Anal Bioanal Chem 398, 2039–2047 (2010). https://doi.org/10.1007/s00216-010-4153-2

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