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Updated: Jun 28, 2026

A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
Published on: April 11, 2014
Fluorimetric determination of ascorbic acid with o-phenylenediamine
Xia Wu1, Yuxia Diao, Changxia Sun
1Key Laboratory for Colloid and Interface Chemistry of Education Ministry, Department of Chemistry, School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, People's Republic of China.
Abstract:
A simple and sensitive fluorimetric method for the determination of ascorbic acid (AA) is described. The method is based on the condensation reaction between AA and o-phenylenediamine (OPDA) in the absence of the oxidant. The fluorescence intensity is measured at excitation and emission wavelengths of 360 and 430 nm, respectively. Under optimum condition, a linear relationship is obtained between the fluorescence intensity and the concentration of AA in the range of 0.05-40 mug ml(-1). The detection limit is 0.006 mug ml(-1), which is obviously lower than that of other fluorimetric methods reported.
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