Analytical Sciences

Development of an Evaluation Method for Hydroxyl Radical Scavenging Activities Using Sequential Injection Analysis with Chemiluminescence Detection

Aoi MIYAMOTO,* Saori NAKANO,** Kaishu NAGAI,** Naoya KISHIKAWA,** Kaname OHYAMA,** Takahiko AOYAMA,* Yoshiaki MATSUMOTO,* and Naotaka KURODA**

*School of Pharmacy, Nihon University, 7-7-1 Narashinodai, Funabashi, Chiba 274-8555, Japan
**Graduate School of Biomedical Sciences, Course of Pharmaceutical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan

A method for evaluating hydroxyl radical (·OH) scavenging activities using sequential injection analysis (SIA) with chemiluminescence (CL) detection was developed. In this system, CL was produced by the reaction of luminol with ·OH generated from the Fenton reaction. The scavenging activity was expressed as a diminution rate of the CL due to the scavenging of ·OH by a sample. The SIA system allows the automation of a series of experimental procedures including Fenton’s reaction, scavenging of ·OH, and luminol CL reaction. The evaluation of scavenging activities in one sample (n = 3) was completed within 3.0 min. Relative standard deviations (n = 3) of scavenging activity with 700 μM L-ascorbic acid were 2.6% (intraday) and 3.7% (interday). The SIA-CL system was applied to measure ·OH scavenging activities of several antioxidants and pharmaceuticals.

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