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Related Experiment Video

Updated: Jun 27, 2026

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

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

Published on: April 11, 2014

Micro determination of ascorbic acid using methyl viologen.

E K Janghel¹, V K Gupta, M K Rai

¹Chhattisgarh Institute of Technology, Near Parri Nala, G.E. Road, Rajnangaon, Chhattisgarh, India.

|December 17, 2008

Summary

A new spectrophotometric method accurately determines ascorbic acid by measuring a blue radical ion. This sensitive technique offers a low detection limit, suitable for analyzing ascorbic acid in various samples.

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Area of Science:

Analytical Chemistry
Spectrophotometry

Background:

Ascorbic acid (vitamin C) is vital for human health.
Accurate quantification of ascorbic acid is crucial in food, pharmaceuticals, and biological studies.
Existing analytical methods may lack sensitivity or simplicity.

Purpose of the Study:

To develop a simple, sensitive, and accurate spectrophotometric method for determining ascorbic acid.
To establish the method's performance characteristics, including sensitivity and detection limits.
To validate the method's applicability to real-world samples.

Main Methods:

A spectrophotometric assay was developed based on the reduction of methyl viologen by ascorbic acid.
The formation of a stable blue free radical ion was measured at 600nm.
The method's linearity, sensitivity, molar absorptivity, and Sandell's sensitivity were determined.

Main Results:

The method demonstrated high sensitivity with a lower limit of detection of 0.1 µg/mL (0.1 ppm) for ascorbic acid.
Beer's law was obeyed in the concentration range of 0.1–1.0 µg/mL.
Molar absorptivity was 1.5x10^5 L/mol/cm, and Sandell's sensitivity was 0.001 µg/cm².
The method showed comparable sensitivity to flow injection analysis.

Conclusions:

The developed spectrophotometric method is simple, sensitive, and accurate for ascorbic acid determination.
The method is suitable for routine analysis of ascorbic acid in diverse matrices.
This technique offers a valuable alternative for quantifying ascorbic acid in food, pharmaceutical, and biological samples.