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Development and validation of an HPTLC–DPPH assay and its application to the analysis of honey

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Abstract

Using honey as a model sample, the aim of this study is to develop and validate a simple, rapid screening tool that allows for the visualization of constituents that contribute to the antioxidant activity of a complex natural product and to quantify their individual antioxidant effects even if their chemical identity is unknown. The method employs a validated analysis, which is based on the separation of constituents using high-performance thin-layer chromatography (HPTLC) followed by their visualization using DPPH* (2,2-diphenyl-1-picrylhydrazyl) as derivatizing reagent and the quantification of the antioxidant activity of individual bands expressed as gallic acid equivalents.

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Acknowledgements

We thank Chromatech Scientific for their technical support of this project.

Funding

This research was supported by funding from the Cooperative Research Centre for Honey Bee Products (Project 13) and the use of samples from AgriFutures Australia (PRJ-010313).

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Authors and Affiliations

  1. Cooperative Research Centre for Honey Bee Products Limited (CRC HBP), Crawley, Western Australia, Australia

    Md Khairul Islam, Katherine Hammer & Cornelia Locher

  2. Division of Pharmacy, School of Allied Health, University of Western Australia, Crawley, Western Australia, 6009, Australia

    Md Khairul Islam, Tomislav Sostaric, Lee Yong Lim & Cornelia Locher

  3. School of Biomedical Sciences, University of Western Australia, Crawley, Western Australia, 6009, Australia

    Katherine Hammer

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  1. Md Khairul Islam
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Correspondence to Cornelia Locher.

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Islam, M.K., Sostaric, T., Lim, L.Y. et al. Development and validation of an HPTLC–DPPH assay and its application to the analysis of honey. JPC-J Planar Chromat 33, 301–311 (2020). https://doi.org/10.1007/s00764-020-00033-0

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  • DOI: https://doi.org/10.1007/s00764-020-00033-0

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