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Updated: Jun 1, 2026

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
On-line detection of antioxidative activity in high-performance liquid chromatography eluates by chemiluminescence
A Dapkevicius1, T A van Beek, H A Niederländer
1Laboratory of Organic Chemistry, Wageningen Agricultural University, Dreijenplein 8, 6703 HB Wageningen, The Netherlands.
Abstract:
Luminol chemiluminescence (CL) was employed for the on-line detection of radical scavengers in HPLC eluates. Optimization of CL reagents and instrumental setup resulted in a steady postcolumn luminol photochemical reaction in the presence of microperoxidase and hydrogen peroxide at pH 10. Quenching of the CL signal was utilized to detect radical scavenging activity of both natural and synthetic antioxidants at the nanogram level. The detection system can be used with isocratic or gradient elution. Several antioxidative compounds were detected in thyme and sage acetone extracts. Quantitative results can be obtained when antioxidants are analyzed at certain concentrations. The method is suitable for rapid screening of antioxidants in crude extracts.
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