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Updated: Aug 20, 2025

A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
Published on: April 11, 2014
Redox-Based Colorimetric Sensor for the Selective Determination of Ascorbic Acid in Fixed-Dose Combination Tablets
Samah F El-Malla1, Fotouh R Mansour1, Rehab H Elattar1
1Tanta University, Medical Campus of Tanta University, Faculty of Pharmacy, Department of Pharmaceutical Analytical Chemistry, El-Geish Street, Tanta 31111, Egypt.
Background:
Ascorbic acid (ASC; vitamin C) is a weak chromophore, so its presence cannot be determined directly by spectrophotometry.
Objective:
This work aims to develop and validate a simple and ecofriendly analytical method for the determination of ASC concentration based on the reaction of ASC with the metal complex, ferric salicylate.
Methods:
The visible absorbance of ferric salicylate was found to be inversely proportional to the concentration of ASC. The possible mechanism of the reaction between ASC and ferric salicylate was investigated: ferric salicylate was found to be reduced by ASC under the applied experimental conditions. Different parameters that may affect the reaction were also investigated: ferric salicylate concentration, pH, ionic strength, and the time of reaction. The optimum concentration of ferric salicylate was found to be 1000 µM and the optimum pH was 5.5. The developed method was validated according to International Conference on Harmonization (ICH) guidelines.
Results:
The linearity range of the developed method was 5-70 µg/mL and the correlation coefficient was 0.9994. The limits of detection and quantitation were 0.38 µg/mL and 1.16 µg/mL, respectively. The method was successfully applied to the determination of ASC concentration in commercial Ruta-C60® tablets. The mean recovery ± standard deviation was found to be 101.10 ± 0.70%. The results were statistically compared to those obtained by a reported HPLC method. Good agreement was observed.
Conclusion:
The developed method is simple, fast, cost-effective, and suitable for routine pharmaceutical analysis of ASC.
Highlights:
The developed method is more sensitive than the other spectrophotometric methods reported for determination of ASC.
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