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Updated: Aug 11, 2026

A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
Published on: April 11, 2014
"Antioxidant" (reducing) efficiency of ascorbate in plasma is not affected by concentration
I F Benzie1, W Y Chung, J J Strain
1Department of Nursing and Health Sciences, The Hong Kong Polytechnic University, Kowloon, Hong Kong SAR, China.
Abstract:
Ascorbate (vitamin C), an important dietary derived antioxidant, reportedly shows decreasing "antioxidant efficiency" with increasing concentrations in indirect radical trapping methods of antioxidant capacity. This study investigated the effect of concentration on antioxidant efficiency of ascorbate using a direct test of antioxidant capacity, the ferric reducing/antioxidant power test (FRAP assay). Results showed that the antioxidant efficiency factor of ascorbate was 2 and was constant over a wide concentration range in both plasma and pure aqueous solution. However, the absolute amount of ascorbate lost per unit of time increased with concentration. Furthermore, ascorbate was less stable in plasma than in aqueous solutions of similar pH and less stable in ethylenediamine tetraacetic acid (EDTA) than in heparinized plasma. Results indicate that previously reported concentration-dependent changes in antioxidant efficiency of ascorbate may have been caused by loss of ascorbate prior to and during testing, and by methodologic characteristics of indirect peroxyl radical trapping tests of antioxidant capacity. Therefore, it is suggested that the premise that the antioxidant efficiency of ascorbate is concentration-dependent is largely methodologically derived and does not reflect the antioxidant behavior of ascorbate per se.
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