Quasi-simultaneous determination of antioxidative activities against superoxide anion and nitric oxide by a combination of sequential injection analysis and flow injection analysis with chemiluminescence detection

Abstract

A method that combines sequential injection analysis (SIA), flow injection analysis and chemiluminescence (CL) detection was developed for the quasi-simultaneous determination of antioxidative activities against superoxide anion (O2-) and nitric oxide (NO). The antioxidative activity was expressed as the decrease in luminol CL intensity caused by the quenching of O2- or NO by an antioxidant. The SIA system consisted of two syringe pumps, two selection valves, two holding coils, an HPLC pump to deliver luminol solution, and a CL detector. Operation of the syringe pumps and multiport valves was controlled automatically using a personal computer with appropriate software. A hypoxanthine (HX)-xanthine oxidase (XOD) system was used for the generation of O2-, and (+/-)-(E)-4-methyl-2-[(E)-hydroxyimino]-5-nitro-6-methoxy-3-hexenamide (NOR1) was employed as NO donor agent. The repeatability of the method was evaluated with 35.2 microg ml(-1) L-ascorbic acid, and the relative standard deviations (RSD) of the antioxidative activities were less than 3.8%. The quasi-simultaneous determination of the antioxidative activities in one sample was completed within 2.0 min. The antioxidative activities of some antioxidants and commercially available supplements containing certain antioxidants were successfully determined using this system. The proposed system is rapid and reproducible, and thus may be useful for the screening of functional foods, supplements and pharmaceutical formulations that exhibit antioxidative activity.

Scavenging of superoxide anion radical and hydroxyl radical by novel thiazolyl‐thiazolidine‐2,4‐dione compounds

“…This reaction is accompanied by a strong light emission [ Fig. 2(A), curve 1], and has become very useful for detecting active oxygen species and evaluating antioxidative effects of drugs (16)(17)(18).…”

Nitric oxide (NO) scavenging capacity of natural antioxidants

“…Although it is well-known that free radicals to be scavenged by antioxidants include nitric oxide (NO), NO has never been a subject for scavenging capacity measurement in ORAC-like methods (Al-Laith, 2010;Cook, Wink, Blount, Krishna, & Hanbauer, 1996;Nishibayashi, Asanuma, Kohno, Gomez-Vargas, & Ogawa, 1996;Poderoso et al, 1999;Rubbo et al, 2000;Sreejayan & Rao, 1997). Recently, using the chemiluminescence-based method with the multisyringe flow injection analysis (MSFIA), the NO scavenging capacity for some antioxidants was evaluated; however the MSFIA method has a few experimental disadvantages such as the low selectivity for NO and the requirement of careful choice of experimental conditions (Miyamoto et al, 2007;Ribeiro, Magalhases, Segundo, Reis, & Lima, 2010). In living biological systems, a high level of NO and its oxidised derivatives such as peroxynitrite are known to be toxic (Szabo & Thiemermann, 1994), but the ability of antioxidants to scavenge NO has been hardly evaluated.…”

A Smart Advanced Chemiluminescence-Sensing Platform for Determination and Imaging of the Tissue Distribution of Natural Antioxidants

“…Antioxidants were separated by a high-performance liquid chromatography (HPLC) system and detected as negative peaks in the chromatogram . Moreover, our research group previously reported the use of luminol CL assays to evaluate antioxidant capacity based on the ability of the antioxidant to scavenge ROS. − However, in these CL assays, the negative signal was recorded against the antioxidant capacity, and the relationship between the CL signal and the antioxidant capacity does not possess good linearity. All of the previously reported antioxidant assays in the literature required the pulverization and homogenization of samples before their analysis.…”

“…Furthermore, during our luminometer-based measurement studies, the compounds with higher antioxidant capacities provided more intense CL. As we mentioned previously, the reported CL methods for evaluation of antioxidants relied mainly on the quenching of CL. − While using quinones as a probe, antioxidant capacity can be interpreted as a positive long-lasting CL signal for the first time through the quinone redox cycle (Figure ).…”