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Updated: Feb 7, 2026

A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
Published on: April 11, 2014
Flow-Injection Chemiluminescence Method for Sensitive Determination of Ascorbic Acid in Fruit Juices and
Tamer H A Hasanin1,2, Terufumi Fujiwara2
1Department of Chemistry, Faculty of Science, Minia University.
Abstract:
A highly sensitive flow-injection (FI) method was developed for the determination of ascorbic acid using chemiluminescence (CL) based detection. This method involved the following processes: (1) reduction of tetrachloroaurate(III) in hydrochloric acid with ascorbic acid; (2) on-line extraction of the residual Au(III) with rhodamine B from the aqueous hydrochloric acid solution into toluene, followed by the separation of the Au(III)-containing organic phase from the aqueous phase through a microporous Teflon membrane in the flow system; and (3) the measurement of CL produced in a flow cell upon mixing of the extract stream of Au(III) in toluene with luminol in the reversed micellar medium of cetyltrimethylammonium chloride-water in 1-hexanol-cyclohexane, which was injected into a CL reagent stream. In this procedure, a reduction in the CL intensity occurred due to the addition of ascorbic acid to the Au(III) solution. The CL signal of Au(III) decreased with increasing concentration of ascorbic acid in the aqueous sample solution. The proposed procedure allowed the indirect quantitative determination of ascorbic acid in the range of 1.0 × 10-12 to 1.0 × 10-7 M with a correlation coefficient of 0.987 and relative standard deviation of 2.1% (n = 6) at 1.0 × 10-9 M. The proposed FI-CL methodology was successfully applied for quantitative determination of ascorbic acid in fruit juices and pharmaceutical samples.
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