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Related Experiment Video

Updated: Jul 15, 2026

A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
11:56

A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells

Published on: April 11, 2014

A simple and sensitive assay for ascorbate using a plate reader.

Jesse M Vislisel1, Freya Q Schafer, Garry R Buettner

  • 1ESR Facility and Free Radical and Radiation Biology, University of Iowa, Iowa City, IA 52242, USA.

Analytical Biochemistry
|April 17, 2007
PubMed
Summary

A new, fast, and affordable plate reader assay accurately measures ascorbate levels in various samples. This method enables high-throughput analysis, significantly reducing experimental time and costs for biological and food sample testing.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Cell Biology

Background:

  • Accurate determination of ascorbate (Vitamin C) is crucial in biological and food science.
  • Existing methods for ascorbate quantification can be time-consuming and costly.
  • Development of a rapid, high-throughput assay is needed for efficient sample analysis.

Purpose of the Study:

  • To develop and validate a rapid, inexpensive, and reliable plate reader-based assay for ascorbate determination.
  • To assess the assay's robustness across diverse sample types.
  • To utilize the assay for investigating ascorbate derivative uptake in cell cultures.

Main Methods:

  • Ascorbic acid is oxidized by Tempol and reacted with o-phenylenediamine.
  • The formation of a fluorescent condensation product is monitored using a plate reader.

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Last Updated: Jul 15, 2026

A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
11:56

A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells

Published on: April 11, 2014

A Rapid and Chemical-free Hemoglobin Assay with Photothermal Angular Light Scattering
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  • The assay was validated using various biological and food samples, including cell lines.

    • Main Results:

    • The assay provides accurate ascorbate quantification in <10 minutes for 96 samples.
    • It demonstrates robustness with diverse matrices like orange juice, plasma, and cultured cells.
    • Investigated ascorbate derivative uptake in U937 cells, revealing differential accumulation rates.

    Conclusions:

    • The developed plate reader assay is a rapid, economical, and reliable tool for ascorbate quantification.
    • It significantly reduces analysis time compared to conventional methods.
    • The assay is suitable for high-throughput screening and biological research applications.