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Paper-based DPPH Assay for Antioxidant Activity Analysis

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Abstract

We report on a paper-based 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl (DPPH) assay for a simple, inexpensive, low reagent and sample consumption and high throughput analysis of antioxidant activity. The paper-based device was fabricated using a lamination method to create a 5-mm in diameter circular test zone that was embedded with a DPPH reagent. The analysis was carried out in one-step by dropping an antioxidant/sample onto the test zone. After reduction by the antioxidant, the DPPH radicals become stable DPPH molecules, resulting in a change in color from deep violet to pale yellow. The violet color intensity of DPPH was inversely proportional to the antioxidant activity of the samples, and was measured using imaging software. A high precision and a low limit of detection were found in the analysis of six standard antioxidants including gallic acid, trolox, ascorbic acid, caffeic acid, vanilliic acid and quercetin. The device was then validated against the traditional spectrophotometric DPPH assay by analyzing the antioxidant activity of 7 tea samples. The results showed no significant difference for gallic acid equivalent for all 7 samples obtained from the two methods at the 95% confidence level, indicating that the developed method was reliable for antioxidant activity analysis of real samples. Finally, the paper-based DPPH device was found to be stable over 10 days when stored in a refrigerator (2–4°C), making it an easy-to-use device for end-users.

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Acknowledgments

This work was financially supported by the Research Grant of Burapha University through National Research Council of Thailand (Grant No. 72/2558 and 93/2559) and the Center of Excellence for Innovation in Chemistry (PERCH-CIC), Commission on Higher Education, Ministry of Education. We would also like to thank Prof. Dr. Federick W. H. Beamish and Prof. Dr. Ron Beckett, Faculty of Science, Burapha University for their comments and correction.

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Author notes

  1. K. S. and S. N. contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Burapha University, Chon Buri, 20131, Thailand

    Kitima Sirivibulkovit, Souksanh Nouanthavong & Yupaporn Sameenoi

  2. Savannakhet Teacher Training College, Savannakhet, China

    Souksanh Nouanthavong

Authors
  1. Kitima Sirivibulkovit
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  2. Souksanh Nouanthavong
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  3. Yupaporn Sameenoi
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Correspondence to Yupaporn Sameenoi.

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Sirivibulkovit, K., Nouanthavong, S. & Sameenoi, Y. Paper-based DPPH Assay for Antioxidant Activity Analysis. ANAL. SCI. 34, 795–800 (2018). https://doi.org/10.2116/analsci.18P014

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  • DOI: https://doi.org/10.2116/analsci.18P014

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