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A simple approach to detect caffeine in tea beverages.

Amit K Ghosh1, Chandrasekhar Ghosh, Ashutosh Gupta

  • 1Unilever R&D Bangalore , 64 Main Road, Whitefield, Bangalore 560066, India.

Journal of Agricultural and Food Chemistry
|April 3, 2013
PubMed
Summary

Acridine Orange (AO) dye detects caffeine in water by fluorescence enhancement. This sensitive method works best at low pH and accurately measures caffeine in tea beverages.

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Area of Science:

  • Analytical Chemistry
  • Biophysical Chemistry

Background:

  • Acridine Orange (AO) is a fluorescent dye existing in monomer-dimer equilibrium in aqueous solutions.
  • The AO monomer is fluorescent, while the dimer is not, forming the basis for fluorescence-based detection.

Purpose of the Study:

  • To develop a selective and sensitive method for caffeine detection using Acridine Orange.
  • To investigate the mechanism of caffeine detection and optimize conditions for enhanced sensitivity.

Main Methods:

  • Utilizing the photophysical properties of Acridine Orange for caffeine sensing.
  • Monitoring shifts in the AO monomer-dimer equilibrium induced by caffeine addition.
  • Evaluating caffeine binding constants at different pH levels (pH ~2 and pH ~7).
  • Comparing the developed method with High-Performance Liquid Chromatography (HPLC) for caffeine quantification in tea beverages.

Main Results:

  • Caffeine addition shifts the AO equilibrium towards the fluorescent monomer, enhancing fluorescence intensity.
  • The enhancement of AO fluorescence serves as a reliable signal for caffeine detection.
  • Caffeine binding to AO monomer is stronger at pH ~2, leading to superior sensitivity compared to pH ~7.
  • The method demonstrated accurate caffeine content evaluation in commercial tea beverages.

Conclusions:

  • Acridine Orange is an effective probe for selective and sensitive caffeine detection in aqueous solutions.
  • Optimizing pH to ~2 significantly enhances caffeine detection sensitivity.
  • The developed AO-based fluorescence method provides a viable alternative for caffeine quantification in beverages.

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