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Paper-based DPPH Assay for Antioxidant Activity Analysis.

Kitima Sirivibulkovit1, Souksanh Nouanthavong1,2, Yupaporn Sameenoi1

  • 1Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Burapha University.

Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry
|July 13, 2018
PubMed
Summary
This summary is machine-generated.

A new paper-based assay simplifies antioxidant activity analysis using the 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl (DPPH) method. This cost-effective device offers reliable results comparable to traditional methods for various samples.

Keywords:
DPPH assayantioxidant activitypaper-based devices

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Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Materials Science

Background:

  • The 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl (DPPH) assay is a standard method for measuring antioxidant activity.
  • Traditional spectrophotometric DPPH assays can be time-consuming and require significant reagent and sample volumes.
  • There is a need for simpler, more accessible, and high-throughput methods for antioxidant analysis.

Purpose of the Study:

  • To develop and validate a novel paper-based 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl (DPPH) assay.
  • To enable simple, inexpensive, and high-throughput analysis of antioxidant activity with minimal reagent and sample consumption.
  • To provide a reliable alternative to traditional spectrophotometric methods for real-world sample analysis.

Main Methods:

  • Fabrication of a paper-based device with an embedded 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl (DPPH) reagent test zone using a lamination method.
  • One-step analysis by applying antioxidant samples to the test zone, observing color change (violet to yellow) upon DPPH reduction.
  • Quantification of antioxidant activity using imaging software based on the inverse relationship between color intensity and activity.
  • Validation against spectrophotometric methods using standard antioxidants and real tea samples.

Main Results:

  • The paper-based 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl (DPPH) assay demonstrated high precision and a low limit of detection for standard antioxidants.
  • Analysis of tea samples showed no significant difference in gallic acid equivalent compared to the traditional spectrophotometric assay (95% confidence level).
  • The developed paper device exhibited stability for over 10 days when stored under refrigeration (2-4°C).

Conclusions:

  • The paper-based 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl (DPPH) assay is a reliable, cost-effective, and user-friendly method for antioxidant activity determination.
  • This innovative approach facilitates high-throughput analysis and reduces reagent and sample requirements.
  • The device is suitable for analyzing antioxidant activity in various real-world samples, offering a practical alternative to conventional methods.